SW846PU32
DETERMINATION OF CARBONYL COMPOUNDS
BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
1.0 SCOPE AND APPLICATION
1.1 This method provides procedures for the determination of free carbonyl
compounds in various matrices by derivatization with 2,4-dinitrophenylhydrazine
(DNPH). The method utilizes high performance liquid chromatography (HPLC) with
ultraviolet/visible (UV/vis) detection to identify and quantitate the target
analytes. This method includes two procedures encompassing all aspects of the
analysis (extraction to determination of concentration). Procedure 1 is
appropriate for the analysis of aqueous, soil and waste samples and stack samples
collected by Method 0011. Procedure 2 is appropriate for the analysis of indoor
air samples collected by Method 0100. The list of target analytes differs by
procedure. The appropriate procedure for each target analyte is listed in the
table below.
Compound CAS No." Proc. lb Proc. 2
b
Acetaldehyde
Acetone
Acrolein
Benzaldehyde
Butanal (butyraldehyde)
Crotonaldehyde
Cyclohexanone
Decanal
2,5-Dimethylbenzaldehyde
Formaldehyde
Heptanal
Hexanal (hexaldehyde)
Isovaleraldehyde
Nonanal
Octanal
Pentanal (valeraldehyde)
Propanal (propionaldehyde)
m-Tolualdehyde
o-Tolualdehyde
p-Tolualdehyde
75-07-0
67-64-1
107-02-8
100-52-7
123-72-8
123-73-9
108-94-1
112-31-2
5779-94-2
50-00-0
111-71-7
66-25-1
590-86-3
124-19-6
124-13-0
110-62-3
123-38-6
620-23-5
529-20-4
104-87-0
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
Chemical Abstract Services Registry Number.
The two procedures have overlapping lists of target compounds that have
been evaluated using modifications of the analysis. Refer to the
respective procedure number when choosing the appropriate analysis
technique for a particular compound.
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1.2 Method detection limits (MDL) using procedure 1 are listed in Tables 1
and 2. Sensitivity data for sampling and analysis by use of Method 0100
(procedure 2) are given in Table 3. The MDL for a specific sample may differ
from the listed value, depending upon the interferences in the sample matrix and
the volume of sample used in the procedure.
1.3 The extraction procedure for solid samples is outlined in Sec. 7.1 of
this method.
1.4 When this method is used to analyze unfamiliar sample matrices,
compound identification should be supported by at least one additional
qualitative technique. A gas chromatograph/mass spectrometer (GC/MS) may be used
for the qualitative confirmation of results for the target analytes, using the
extract produced by this method.
1.5 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of chromatography and in the interpretation of
chromatograms. Each analyst must demonstrate the ability to generate acceptable
results with this method.
2.0 SUMMARY OF METHOD
This method contains two procedures dealing with different sample types.
2.1 Liquid and Solid Samples (Procedure 1)
2.1.1 For wastes comprised of solids, and aqueous wastes containing
greater than one percent solid material, the aqueous phase should be
separated from the solid phase and stored, according to Sec. 6.2, for
possible later analysis. If necessary, the particle size of the solids in
the waste is reduced. The solid phase is extracted with a volume of fluid
equal to 20 times the sample's weight. The extraction fluid employed is
a function of the alkalinity of the solid phase of the waste. A special
extractor is used when volatiles are being extracted. Following
extraction, the extract is filtered through a 0.6 - 0.8 /zm glass fiber
filter.
2.1.2 If compatible (i.e., multiple phases will not form on
combination), the initial aqueous phase of the waste is added to the
aqueous extract, and these liquids are analyzed together. If incompatible,
the liquids are analyzed separately and the results are mathematically
combined to yield a volume-weighted average concentration.
2.1.3 A measured volume of aqueous sample (approx. 100 mL) or an
appropriate amount of solids extract (approx. 25 g), is buffered to pH 3
and derivatized with 2,4-dinitrophenylhydrazine (DNPH), using either the
appropriate solid-phase or a liquid-liquid extraction technique. If the
solid-phase extraction (SPE) option is used, the derivatized compound is
extracted using solid sorbent cartridges, then eluted with ethanol. If the
liquid-liquid option is used, the derivatized compound is serially
extracted three (3) times with methylene chloride. The methylene chloride
extracts are concentrated using the appropriate procedure 3500 series
method and exchanged with acetonitrile prior to HPLC analysis. HPLC
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conditions are described which permit the separation and measurement of
various carbonyl compounds in the extract by absorbance detection at 360
nm.
2.1.4 If formaldehyde is the only analyte of interest, the aqueous
sample and/or solid sample extract should be buffered to pH 5.0 to minimize
the formation of artifact formaldehyde.
2.2 Stack Gas Samples Collected by Method 0011 (Procedure 1): The entire
sample returned to the laboratory is extracted with methylene chloride and the
extract is diluted or concentrated to a known volume. An aliquot of the
methylene chloride extract is solvent exchanged and concentrated or diluted as
necessary. HPLC conditions are described that permit the separation and
measurement of various carbonyl compounds in the extract by absorbance detection
at 360 nm.
2.3 Indoor Air Samples by Method 0100 (Procedure 2): The sample
cartridges are returned to the laboratory and backflushed with acetonitrile into
a 5-mL volumetric flask. The eluate is diluted to volume with acetonitrile. Two
aliquots of the acetonitrile extract are pipetted into two sample vials having
Teflon®-!ined septa. HPLC conditions are described which allow for the
separation and measurement of the various carbonyl compounds in the extract by
absorbance detection at 360 nm.
3.0 INTERFERENCES
3.1 Method interferences may be caused by contaminated solvents, reagents,
glassware, and other sample processing hardware which can lead to discrete
artifacts and/or elevated chromatogram baselines. All materials should routinely
demonstrate freedom from interferences under analysis conditions by analyzing
laboratory reagent blanks as described in Sec. 8.5.
3.1.1 Glassware must be scrupulously cleaned. Glassware should be
rinsed as soon as possible after use 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. After washing the glassware should
then be drained, dried, and heated in a laboratory oven at 130"C for two
to three hours before reuse. Solvent rinses with acetonitrile may be
substituted for the oven heating. After drying and cooling, glassware
should be stored in a clean environment to prevent accumulation of dust or
other contaminants.
NOTE: Do not rinse glassware with acetone or methanol. These solvents react
with DNPH to form interferences.
3.1.2 The use of high purity reagents and solvents helps minimize
interference. Purification of solvents by distillation in all glass
systems may be required.
3.1.3 Polyethylene gloves must be worn when handling silica gel
cartridges to reduce the possibility of contamination.
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3.2 Formaldehyde contamination of the DNPH reagent is frequently
encountered due to its widespread occurrence in the environment. The DNPH
reagent in Procedure 2 must be purified by multiple recrystallizations in HPLC-
grade acetonitrile. Recrystallization is accomplished, at 40 - 60°C, by slow
evaporation of the solvent to maximize crystal size. The purified DNPH crystals
are stored under HPLC-grade acetonitrile. Impurity levels of carbonyl compounds
in the DNPH are determined prior to sample analysis and should be less than 25
mg/L. Refer to Appendix A for the recrystallization procedure.
3.3 Matrix interferences may be caused by contaminants co-extracted from
the sample. The extent of matrix interferences will be source- and matrix-
specific. If interferences occur in subsequent samples, modification of the
mobile phase or some additional cleanup may be necessary.
3.4 In Procedure 1, acetaldehyde is generated during the derivatization
step if ethanol is present in the sample. This background will impair the
measurement of acetaldehyde levels below 0.5 ppm (500 ppb).
3.5 For Procedure 2, at the stated two column analysis conditions, the
identification and quantitation of butyraldehyde may be difficult due to
coelution with isobutyraldehyde and methyl ethyl ketone. Precautions should be
taken and adjustment of the analysis conditions should be made to avoid potential
problems.
4.0 APPARATUS AND MATERIALS
4.1 High performance liquid chromatograph (modular).
4.1.1 Pumping system - Gradient, with constant flow control capable
of 1.50 mL/min.
4.1.2 High pressure injection valve with 20-juL loop.
4.1.3 Column - 250 mm x 4.6 mm ID, 5-jiim particle size, CIS (Zorbax
or equivalent).
4.1.4 Absorbance detector - 360 nm.
4.1.5 Strip-chart recorder compatible with detector - Use of a data
system for measuring peak areas and retention times is recommended.
4.1.6 Helium - for degassing mobile phase solvents. (Procedures
1 and 2)
4.1.7 Mobile phase reservoirs and suction filtration apparatus - For
holding and filtering HPLC mobile phase. Filtering system should be all
glass and Teflon® and use a 0.22 ;um polyester membrane filter.
4.1.8 Syringes - for HPLC injection loop loading, with capacity at
least four times the loop volume.
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4.2 Apparatus and materials for Procedure 1
4.2.1 Reaction vessel - 250-mL Florence flask.
4.2.2 Separatory funnel - 250-mL, with Teflon® stopcock.
4.2.3 Kuderna-Danish (K-D) apparatus - See Method 3510 or other
appropriate 3500 series method. (Other concentration apparatus may be
employed if the laboratory can demonstrate equivalent performance).
4.2.4 Boiling chips - Solvent-extracted with methylene chloride,
approximately 10/40 mesh (silicon carbide or equivalent).
4.2.5 pH meter - Capable of measuring to 0.01 pH units.
4.2.6 Glass fiber filter paper - 1.2 /zm pore size (Fisher Grade G4
or equivalent).
4.2.7 Solid sorbent cartridges - Packed with 2 g CIS (Baker or
equivalent).
4.2.8 Vacuum manifold - Capable of simultaneous extraction of up to
12 samples (Supelco or equivalent).
4.2.9 Sample reservoirs - 60-mL capacity (Supelco or equivalent).
4.2.10 Pipet - Capable of accurately delivering 0.10 ml solution.
4.2.11 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 2°C). The bath should be used in a hood.
4.2.12 Sample shaker - Controlled temperature incubator (± 2°C) with
orbital shaking (Lab-Line Orbit Environ-Shaker Model 3527 or equivalent).
4.2.13 Syringes - 5-mL, 500-/iL, 100-^L, (Luer-Lok or equivalent).
4.2.14 Syringe filters - 0.45 p.m filtration disks (Gelman Acrodisc
4438 or equivalent).
4.3 Apparatus and materials for Procedure 2
4.3.1 Syringes - 10-mL, with Luer-Lok type adapter, used to
backflush the sample cartridges by gravity feed.
4.3.2 Syringe rack - made of an aluminum plate with adjustable legs
on all four corners. Circular holes of a diameter slightly larger than the
diameter of the 10-mL syringes are drilled through the plate to allow batch
processing of cartridges for cleaning, coating, and sample elution. A
plate (0.16 x 36 x 53 cm) with 45 holes in a 5 x 9 matrix is recommended.
See Figure 2 in Method 0100.
4.4 Volumetric flasks - 5-mL, 10-mL, and 250- or 500-mL.
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4.5 Vials - 10- or 25-mL, glass with Teflon®-lined screw caps or crimp
tops.
4.6 Balance - Analytical, capable of accurately weighing to 0.0001 g.
4.7 Glass funnels
4.8 Polyethylene gloves - used to handle silica gel cartridges.
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 - Water in which an interferant is not
observed at the method detection limit for the compounds of interest.
5.3 Formalin - Solution of formaldehyde (CH20) in organic-free reagent
water, nominally 37.6 percent (w/w). Exact concentration will be determined for
the stock solution in Sec. 5.7.1.1.
5.4 Aldehydes and ketones - analytical grade, used for preparation of DNPH
derivative standards of target analytes other than formaldehyde. Refer to the
target analyte list.
5.5 Procedure 1 reagents
5.5.1 Methylene chloride, CH2C12 - HPLC grade or equivalent.
5.5.2 Acetonitrile, CH3CN - HPLC grade or equivalent.
5.5.3 Sodium hydroxide solutions, NaOH, 1.0 N and 5 N.
5.5.4 Sodium chloride, NaCl, saturated solution - Prepare by
dissolving an excess of reagent grade solid in organic-free reagent water.
5.5.5 Sodium sulfite solution, Na2S03, 0.1 M.
5.5.6 Sodium sulfate, Na2S04 - granular, anhydrous.
5.5.7 Citric acid, C8H807, 1.0 M solution.
5.5.8 Sodium citrate, C6H5Na307«2H20, 1.0 M trisodium salt dihydrate
solution.
5.5.9 Acetic acid (glacial), CH3C02H.
5.5.10 Sodium acetate, CH3C02Na.
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5.5.11 Hydrochloric acid, HC1, 0.1 N.
5.5.12 Citrate buffer, 1 M, pH 3 - Prepare by adding 80 ml of 1 M
citric acid solution to 20 mL of 1 M sodium citrate solution. Mix
thoroughly. Adjust pH with NaOH or HC1 as needed.
5.5.13 pH 5.0 Acetate buffer (5M) - Formaldehyde analysis only.
Prepared by adding 40 ml 5M acetic acid solution to 60 ml 5M sodium acetate
solution. Mix thoroughly. Adjust pH with NaOH or HC1 as needed.
5.5.14 2,4-Dinitrophenylhydrazine, 2,4-(02N)2C6H3]NHNH2, (DNPH), 70%
in organic-free reagent water (w/w). Prepare a 3.00 mg/mL solution by
dissolving 428.7 mg of 70% (w/w) DNPH solution in 100 ml of acetonitrile.
5.5.15 Extraction fluid for Procedure 1 - Dilute 64.3 mL of 1.0 N
NaOH and 5.7 ml glacial acetic acid to 900 ml with organic-free reagent
water. Dilute to 1 liter with organic-free reagent water. The pH should
be 4.93 ± 0.02.
5.6 Procedure 2 reagents
5.6.1 Acetonitrile, CH3CN - HPLC grade.
5.6.2 2,4-Dinitrophenylhydrazine, C6HaN404, (DNPH) - recrystallize
at least twice with HPLC-grade acetonitrile using procedure in Appendix A.
5.7 Stock standard solutions for Procedure 1
5.7.1 Stock formaldehyde (approximately 1000 mg/L) - Prepare by
diluting an appropriate amount of the certified or standardized
formaldehyde (approximately 265 /zL) to 100 mL with organic-free reagent
water. If a certified formaldehyde solution is not available or there is
any question regarding the quality of a certified solution, the solution
may be standardized using the procedure in Sec. 5.7.1.1.
5.7.1.1 Standardization of formaldehyde stock solution -
Transfer a 25 mL aliquot of a 0.1 M Na2S03 solution to a beaker and
record the pH. Add a 25.0 mL aliquot of the formaldehyde stock
solution (Sec. 5.18.1) and record the pH. Titrate this mixture back
to the original pH using 0.1 N HC1. The formaldehyde concentration
is calculated using the following equation:
* *• , /, x (30.03)(N HCl)(mL HC1)
Concentration (mg/L) = - — — L
25.0 mL
where:
N HC1 = Normality of HC1 solution used (in milli-equivalents/mL)
(1 mmole of HC1 = 1 milli-equivalent of HC1)
mL HC1 = mL of standardized HC1 solution used
30.03 = Molecular of weight of formaldehyde (in mg/mmole)
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5.7.2 Stock aldehyde(s) and ketone(s) - Prepare by adding an
appropriate amount of the pure material to 90 ml of acetonitrile and dilute
to 100 ml, to give a final concentration of 1000 mg/L.
5.8 Stock standard solutions for Procedure 2
5.8.1 Preparation of the DNPH Derivatives for HPLC analysis
5.8.1.1 To a portion of the recrystallized DNPH, add
sufficient 2N HC1 to obtain an approximately saturated solution. Add
to this solution the target analyte in molar excess of the DNPH.
Filter the DNPH derivative precipitate, wash it with 2N HC1, wash it
again with water, and allow it to dry in air.
5.8.1.2 Check the purity of the DNPH derivative by melting
point determination or HPLC analysis. If the impurity level is not
acceptable, recrystallize the derivative in acetonitrile. Repeat the
purity check and recrystallization as necessary until 99% purity is
achieved.
5.8.2 Preparation of DNPH derivative standards and calibration
standards for HPLC analysis
5.8.2.1 Stock standard solutions - Prepare individual stock
standard solutions for each of the target analyte DNPH derivatives by
dissolving accurately weighed amounts in acetonitrile. Individual
stock solutions of approximately 100 mg/L may be prepared by
dissolving 0.010 g of the solid derivative in 100 mL of acetonitrile.
5.8.2.2 Secondary dilution standard(s) - Using the
individual stock standard solutions, prepare secondary dilution
standards in acetonitrile containing the DNPH derivatives from the
target analytes mixed together. Solutions of 100 jug/L may be
prepared by placing 100 /iL of a 100 mg/L solution in a 100 mL
volumetric flask and diluting to the mark with acetonitrile.
5.8.2.3 Calibration standards - Prepare a working
calibration standard mix from the secondary dilution standard, using
the mixture of DNPH derivatives at concentrations of 0.5 - 2.0 M9/L
(which spans the concentration of interest for most indoor air work).
The concentration of the DNPH derivative in the standard mix solutions
may need to be adjusted to reflect relative concentration distribution
in a real sample.
5.9 Standard storage - Store all standard solutions at 4°C in a glass vial
with a Teflon®-!ined cap, leaving minimum headspace, and in the dark. Standards
should be stable for about 6 weeks. All standards should be checked frequently
for signs of degradation or evaporation, especially just prior to preparing
calibration standards from them.
5.10 Calibration standards
Prepare calibration solutions at a minimum of 5 concentrations for
each analyte of interest in organic-free reagent water (or acetonitrile for
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Procedure 2) from the stock standard solution. The lowest concentration
of each analyte should be at, or just above, the MDLs listed in Tables 1
or 2. The other concentrations of the calibration curve should correspond
to the expected range of concentrations found in real samples.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes, Sec.
4.1.
6.2 Samples must be refrigerated at 4°C. Aqueous samples must be
derivatized and extracted within 3 days of sample collection. The holding times
of leachates of solid samples should be kept at a minimum. All derivatized
sample extracts should be analyzed within 3 days after preparation.
6.3 Samples collected by Methods 0011 or 0100 must be refrigerated at
4*C. It is recommended that samples be extracted and analyzed within 30 days of
collection.
7.0 PROCEDURE
7.1 Extraction of solid samples (Procedure 1)
7.1.1 All solid samples should be made as homogeneous as possible
by stirring and removal of sticks, rocks, and other extraneous material.
When the sample is not dry, determine the dry weight of the sample, using
a representative aliquot. If particle size reduction is necessary, proceed
as per Method 1311.
7.1.1.1 Determination of dry weight - In certain cases,
sample results are desired based on a dry weight basis. When such
data are desired or required, a portion of sample for dry weight
determination should be weighed out at the same time as the portion
used for analytical determination.
WARNING; The drying oven should be contained in a hood or vented. Significant
laboratory contamination may result from drying a heavily contaminated
hazardous waste sample.
7.1.1.2 Immediately after weighing the sample for
extraction, weigh 5 - 10 g of the sample into a tared crucible.
Determine the % dry weight of the sample by drying overnight at
105°C. Allow to cool in a desiccator before weighing:
<,/ j • Li sample ...
% dry weight = - - - - 1 — xlOO
g of sample
7.1.2 Measure 25 g of solid into a 500-mL bottle with a Teflon®-
lined screw cap or crimp top, and add 500 mL of extraction fluid (Sec.
5.5.15). Extract the solid by rotating the bottle at approximately 30 rpm
for 18 hours. Filter the extract through glass fiber filter paper and
store in sealed bottles at 4°C. Each mL of extract represents 0.050 g
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solid. Smaller quantities of solid sample may be used with correspondingly
reduced volumes of extraction fluid maintaining the 1:20 mass to volume
ratio.
7.2 Cleanup and separation (Procedure 1)
7.2.1 Cleanup procedures may not be necessary for a relatively clean
sample matrix. The cleanup procedures recommended in this method have been
used for the analysis of various sample types. If particular samples
demand the use of an alternative cleanup procedure, the analyst must
determine the elution profile and demonstrate that the recovery of
formaldehyde from a spiked sample is greater than 85%. Recovery may be
lower for samples which form emulsions.
7.2.2 If the sample is not clear, or the complexity is unknown, the
entire sample should be centrifuged at 2500 rpm for 10 minutes. Decant the
supernatant liquid from the centrifuge bottle, and filter through glass
fiber filter paper into a container which can be tightly sealed.
7.3 Derivatization (Procedure 1)
7.3.1 For aqueous samples, measure an aliquot of sample which is
appropriate to the anticipated analyte concentration range (nominally
100 mL). Quantitatively transfer the sample aliquot to the reaction vessel
(Sec. 4.2).
7.3.2 For solid samples, 1 to 10 ml of extract (Sec. 7.1) will
usually be required. The amount used for a particular sample must be
determined through preliminary experiments.
NOTE: In cases where the selected sample or extract volume is less than 100 ml,
the total volume of the aqueous layer should be adjusted to 100 ml with
organic-free reagent water. Record original sample volume prior to
dilution.
7.3.3 Derivatization and extraction of the target analytes may be
accomplished using the liquid-solid (Sec. 7.3.4) or liquid-liquid (Sec.
7.3.5) procedures.
7.3.4 Liquid-solid derivatization and extraction
7.3.4.1 For analytes other than formaldehyde, add 4 ml of
citrate buffer and adjust the pH to 3.0 ± 0.1 with 6M HC1 or 6M NaOH.
Add 6 mL of DNPH reagent, seal the container, and place in a heated
(40eC), orbital shaker (Sec. 4.2.12) for 1 hour. Adjust the agitation
to produce a gentle swirling of the reaction solution.
7.3.4.2 If formaldehyde is the only analyte of interest, add
4 mL acetate buffer and adjust pH to 5.0 ± 0.1 with 6M HC1 or 6M NaOH.
Add 6 mL of DNPH reagent, seal the container, and place in a heated
(40eC), orbital shaker (Sec. 4.2.12) for 1 hour. Adjust the agitation
to produce a gentle swirling of the reaction solution.
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7.3.4.3 Assemble the vacuum manifold and connect to a water
aspirator or vacuum pump. Attach a 2-g sorbent cartridge to the
vacuum manifold. Condition each cartridge by passing 10 mL dilute
citrate buffer (10 ml of 1 M citrate buffer dissolved in 250 mL of
organic-free reagent water) through each sorbent cartridge.
7.3.4.4 Remove the reaction vessel from the shaker
immediately at the end of the one hour reaction period and add 10 ml
saturated NaCl solution to the vessel.
7.3.4.5 Quantitatively transfer the reaction solution to the
sorbent cartridge and apply a vacuum so that the solution is drawn
through the cartridge at a rate of 3 to 5 mL/min. Continue applying
the vacuum for about 1 minute after the liquid sample has passed
through the cartridge.
7.3.4.6 While maintaining the vacuum conditions described in
Sec. 7.3.4.4, elute each cartridge train with approximately 9 mL of
acetonitrile directly into a 10 mL volumetric flask. Dilute the
solution to volume with acetonitrile, mix thoroughly, and place in a
tightly sealed vial until analyzed.
NOTE: Because this method uses an excess of DNPH, the cartridges will remain a
yellow color after completion of Sec. 7.3.4.5. The presence of this color
is not indicative of the loss of the analyte derivatives.
7.3.5 Liquid-liquid derivatization and extraction
7.3.5.1 For analytes other than formaldehyde, add 4 mL of
citrate buffer and adjust the pH to 3.0 ± 0.1 with 6M HC1 or 6M NaOH.
Add 6 mL of DNPH reagent, seal the container, and place in a heated
(40°C), orbital shaker for 1 hour. Adjust the agitation to produce
a gentle swirling of the reaction solution.
7.3.5.2 If formaldehyde is the only analyte of interest, add
4 mL acetate buffer and adjust pH to 5.0 ± 0.1 with 6M HC1 or 6M NaOH.
Add 6 mL of DNPH reagent, seal the container, and place in a heated
(40°C), orbital shaker for 1 hour. Adjust the agitation to produce
a gentle swirling of the reaction solution.
7.3.5.3 Serially extract the solution with three 20 mL
portions of methylene chloride using a 250 mL separatory funnel. If
an emulsion forms upon extraction, remove the entire emulsion and
centrifuge at 2000 rpm for 10 minutes. Separate the layers and
proceed with the next extraction. Combine the methylene chloride
layers in a 125-mL Erlenmeyer flask containing 5.0 grams of anhydrous
sodium sulfate. Swirl contents to complete the extract drying
process.
7.3.5.4 Assemble a Kuderna-Danish (K-D) concentrator by
attaching a 10-mL concentrator tube to a 500-mL evaporator flask. See
Sec. 4.0 of Method 3510. Pour the extract into the evaporator flask
being careful to minimize transfer of sodium sulfate granules. Wash
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the Erlenmeyer flask with 30 ml of methylene chloride and add wash to
the evaporator flask to complete quantitative transfer.
7.3.5.5 Concentrate the extract to a final volume of 5 ml,
using the K-D techniques, as described in Method 3510. Exchange the
solvent to acetonitrile prior to analysis.
7.4 Extraction of samples from Methods 0011 and 0100 (Procedures 1 and 2)
7.4.1 Stack gas samples collected by Method 0011 (Procedure 1)
7.4.1.1 Measure the volume of the aqueous phase of the
sample prior to extraction (for moisture determination when the volume
was not measured in the field). Pour the sample into a separatory
funnel and drain the methylene chloride into a volumetric flask.
7.4.1.2 Extract the aqueous solution with two or three
aliquots of methylene chloride. Add the methylene chloride extracts
to the volumetric flask.
7.4.1.3 Fill the volumetric flask to the line with methylene
chloride. Mix well and remove an aliquot.
7.4.1.4 If high concentrations of formaldehyde are present,
the extract can be diluted with mobile phase, otherwise the extract
solvent must be exchanged as described in Sec. 7.3.5.5. If low
concentrations of formaldehyde are present, the sample should be
concentrated during the solvent exchange procedure.
7.4.1.5 Store the sample at 4°C. If the extract will be
stored longer than two days, it should be transferred to a vial with
a Teflon®-! ined screw cap, or a crimp top with a Teflon®-! ined septum.
Proceed with HPLC chromatographic analysis if further cleanup is not
required.
7.4.2 Ambient air samples collected by Method 0100 (Procedure 2)
7.4.2.1 The samples will be received by the laboratory in a
friction-top can containing 2 - 5 cm of granular charcoal, and should
be stored in this can, in a refrigerator, until analysis.
Alternatively, the samples may also be stored alone in their
individual glass containers. The time between sampling and analysis
should not exceed 30 days.
7.4.2.2 Remove the sample cartridge from the labeled culture
tube. Connect the sample cartridge (outlet or long end during
sampling) to a clean syringe.
NOTE: The liquid flow during desorption should be in the opposite direction from
the air flow during sample collection (i.e, backflush the cartridge).
7.4.2.3 Place the cartridge/syringe in the syringe rack.
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7.4.2.4 Backflush the cartridge (gravity feed) by passing
6 ml of acetonitrile from the syringe through the cartridge to a
graduated test tube, or to a 5-mL volumetric flask.
NOTE: A dry cartridge has an acetonitrile holdup volume slightly greater than 1
ml. The eluate flow may stop before the acetonitrile in the syringe is
completely drained into the cartridge because of air trapped between the
cartridge filter and the syringe Luer-Lok tip. If this happens, displace
the trapped air with the acetonitrile in the syringe using a long-tip
disposable Pasteur pipet.
7.4.2.5 Dilute to the 5 ml mark with acetonitrile. Label
the flask with sample identification. Pipet two aliquots into sample
vials having Teflon®-!ined septa.
7.4.2.6 Store the sample at 4°C. Proceed with HPLC
chromatographic analysis of the first aliquot if further cleanup is
not required. Store the second aliquot in the refrigerator until the
results of the first aliquot analysis are complete and validated. The
second aliquot can be used for confirmatory analysis, if necessary.
7.5 Recommended chromatographic conditions
7.5.1 Procedure 1 - For aqueous samples, soil or waste samples, and
stack gas samples collected by Method 0011.
Column:
Mobile Phase Gradient:
Flow Rate:
Detector:
Injection Volume:
CIS, 4.6 mm x 250 mm ID, 5 jum particle size
70/30 acetonitrile/water (v/v), hold for 20 min.
70/30 acetonitrile/water to 100% acetonitrile in
15 min.
100% acetonitrile for 15 min.
1.2 mL/min
Ultraviolet, operated at 360 nm
20 ML
7.5.2 Procedure 2 - For ambient air samples collected by Method
0100.
Column:
Mobile Phase Gradient:
Two HPLC columns, 4.6 mm x 250 mm ID, (Zorbax
CDS, or equivalent) in series
60/40 CH3CN/H20, hold for 0 min.
60/40 to 75/25 CH3CN/H20, linearly in 30 min.
75/25 to 100% CH3CN, linearly in 20 min.
100% CH3CN for 5 minutes.
100% to 60/40 CH3CN/H20, linearly in 1 min.
60/40 CH3CN/H20 for 15 minutes.
Detector:
Ultraviolet, operated at 360 nm
8315A - 13
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Flow Rate: 1.0 mL/min
Sample Injection volume: 25 fj.1 (suggested)
NOTE: Analysts are advised to adjust their HPLC systems to optimize
chromatographic conditions for their particular analytical needs. The
separation of acrolein, acetone, and propionaldehyde should be a minimum
criterion of the optimization in Procedure 2.
7.5.3 Filter and degas the mobile phase to remove dissolved gasses,
using the following procedure:
7.5.3.1 Filter each solvent (water and acetonitrile) through
a 0.22 jum polyester membrane filter, in an all glass and Teflon®
suction filtration apparatus.
7.5.3.2 Degas each filtered solution by purging with helium
for 10 - 15 minutes (100 mL/min) or by heating to 60°C for 5 - 10
minutes in an Erlenmeyer flask covered with a watch glass. A constant
back pressure restrictor (350 kPa) or 15 - 30 cm of 0.25 mm ID Teflon®
tubing should be placed after the detector to eliminate further mobile
phase outgassing.
7.5.3.3 Place the mobile phase components in their
respective HPLC solvent reservoirs, and program the gradient system
according to the conditions listed in Sec. 7.5.2. Allow the system
to pump for 20 - 30 minutes at a flow rate of 1.0 mL/min with the
initial solvent mixture ratio (60%/40% CH3CN/H20). Display the
detector output on a strip chart recorder or similar output device to
establish a stable baseline.
7.6 Calibration
7.6.1 Establish liquid chromatographic operating conditions to
produce a retention time similar to that indicated in Table 1 for the
liquid-solid derivatization and extraction or in Table 2 for liquid-liquid
derivatization and extraction. For determination of retention time
windows, see Sec. 7.5 of Method 8000. Suggested chromatographic conditions
are provided in Sec. 7.5.
7.6.2 Process each calibration standard solution through
derivatization and extraction, using the same procedure employed for sample
processing (Sees. 7.3.4 or 7.3.5).
7.6.3 Analyze a solvent blank to ensure that the system is clean and
interference free.
NOTE: The samples and standards must be allowed to come to ambient temperature
before analysis.
7.6.4 Analyze each processed calibration standard using the
chromatographic conditions listed in Sec. 7.5, and tabulate peak area
against calibration solution concentration in
8315A - 14 Revision 1
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7.6.5 Tabulate the peak area along with standard concentration
injected to determine the calibration factor (CF) for the analyte at each
concentration (see Sec. 7.8.1 for equations). The percent relative
standard deviation (%RSD) of the mean CF of the calibration standards
should be < 20 percent or a system check will have to be performed. If a
calibration check after the system check does not meet the criteria, a
recalibration will have to be performed. If the recalibration does not
meet the established criteria, new calibration standards must be made.
7.6.6 The working calibration curve must be verified each day,
before and after analyses are performed, by analyzing one or more
calibration standards. The calibration factor obtained should fall within
± 15 percent of the initially established calibration factor or a system
check will have to be performed. If a calibration check after the system
check does not meet the criteria, the system must be recalibrated.
7.6.7 After 10 sample runs, or less, one of the calibration
standards must be reanalyzed to ensure that the DNPH derivative calibration
factors remain within ± 15% of the original calibration factors.
7.7 Sample analysis
7.7.1 Analyze samples by HPLC, using conditions established in Sec.
7.5. For Procedure 1 analytes, Tables 1 and 2 list the retention times and
MDLs that were obtained under these conditions. For Procedure 2 analytes,
refer to Figure 3 for the sample chromatogram.
7.7.2 If the peak area exceeds the linear range of the calibration
curve, a smaller sample injection volume should be used. Alternatively,
the final solution may be diluted with acetonitrile and reanalyzed.
7.7.3 After elution of the target analytes, calculate the
concentration of analytes found in the samples using the equations found
in Sec. 7.8 or the specific sampling method used.
7.7.4 If the peak area measurement is prevented by the presence of
observed interferences, further cleanup is required.
7.8 Calculations
7.8.1 Calculate each calibration factor, mean calibration factor,
standard deviation, and percent relative standard deviation as follows:
_ _ Peak Area of the Compound in the Standard
Concentration of the Compound Injected (in ug/L)
8315A - 15 Revision 1
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mean CF = CF =
SD =
V(CF.-CF)2
^ ' SD
RSD = — x 100
n-1 CF
where:
CF * Mean calibration factor using the 5 calibration concentrations.
CFj = Calibration factor for calibration standard i (i = 1-5).
RSD = Relative standard deviation of the calibration factors.
n = Number of calibration standards.
7.8.2 Calculate the concentrations in liquid samples as follows:
Concentration of aldehydes (ug/L) = (Area of sample peak) x 100
IT x Vs
where:
CF = Mean calibration factor the analyte.
Vs = Number of ml of sample (unitless).
7.8.3 Calculate the concentration in solid samples as follows:
Concentration of aldehydes (ug/g) = (Area of sample peak) x 100
where:
CF = Mean calibration factor the analyte.
Vex = Number of ml extraction fluid aliquot (unitless).
7.8.4 Calculate the concentration of formaldehyde in stack gas
samples (Method 0011) as follows: (Procedure 1)
7.8.4.1 Calculation of total formaldehyde
To determine the total formaldehyde in mg, use the following
equation:
-r . , f -,... ,. „ nr (g/mole formaldehyde) ...3 ,
Total mg formaldehyde = C. x V x DF x—— 1—'.— x 10 d mg/ug
d (g/mole DNPH derivatiibe)
8315A - 16 Revision 1
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where:
Cd = measured concentration of DNPH-formaldehyde derivative, mg/L
V = organic extract volume, ml
DF = dilution factor
7.8.4.2 Formaldehyde concentration in stack gas
Determine the formaldehyde concentration in the stack gas
using the following equation:
_ K x (total formaldehyde in mg)
m(std)
where:
K
"mlstd)
= 35.31 ft3/")3, if Vm(std) is expressed in English units
= 1.00 m3/m3, if Vm(8td) is expressed in metric units
= volume of gas sample as measured by dry gas meter,
corrected to standard conditions, dscm (dscf)
7.8.5 Calculation of the concentration of formaldehyde and other
carbonyls from indoor air sampling by Method 0100. (Procedure 2)
7.8.5.1 The concentration of target analyte "a," in air at
standard conditions (25°C and 101.3 kPa), Conc8td in ng/L, may be
calculated using the following equation:
.n _ (Areaa)(Vo1a)(MWa)(1000 ng/ug) ^ ^
(RF)(MWd)(VTotStd )(1000inl/L)
where:
Areaa
CF
Vol.
MWa
MWd
"TotStd
DF
"a"
from the
in
Area of the sample peak for analyte "a"
Mean calibration factor for analyte
calibration in M9/L. (See Sec. 7.8.1)
Total volume of the sample cartridge eluate (ml)
Molecular weight of analyte "a" in g/mole
Molecular weight of the DNPH derivative of analyte
g/mole
Total volume of air sampled converted to standard
conditions in liters (L). (To calculate the concentration
at sampling conditions use Vtot.) (See Sec. 9.1.3 of Method
0100)
Dilution Factor for the sample cartridge eluate, if any.
If there is no dilution, DF = 1
8315A - 17
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7.8.5.2 The target analyte "a" concentration at standard
conditions may be converted to parts per billion by volume, Conca in
ppbv, using the following equation:
Conca ,in ppbv =
(Conco_,td)(22.4)
(MM.)
where:
C°nca.std = Concentration of "a," at standard conditions, in ng/L
22.4 = Ideal gas law volume (22.4 nl_ of gas = 1 nmole, at
standard conditions)
MWa = Molecular weight of analyte "a" in g/mole (or ng/nmole)
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 laboratory should also maintain records to document the quality of the data
generated.
8.2 Quality control procedures necessary to evaluate the HPLC 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, 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.
8315A - 18 Revision 1
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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 Refer to Table 4 for QC acceptance limits derived from the
interlaboratory method validation study on Method 8315.
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 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 MDLs for Procedure 1 listed in Table 1 were obtained using
organic-free reagent water and liquid-solid extraction. The MDLs for Procedure
1 listed in Table 2 were obtained using organic-free reagent water and methylene
chloride extraction. Results reported in Tables 1 and 2 were achieved using
fortified reagent water volumes of 100 mL. Lower detection limits may be
obtained using larger sample volumes.
9.1.1 Procedure 1 of this method has been tested for linearity of
recovery from spiked organic-free reagent water and has been demonstrated
to be applicable over the range 50-1000 jug/L .
9.1.2 To generate the MDL and precision and accuracy data reported
in this section, analytes were segregated into two spiking groups, A and
B. Chromatograms using liquid-solid and liquid-liquid extraction are
presented in Figures 1 (a and b) and 2 (a and b), respectively.
9.2 The sensitivity of Procedure 2 sampling (Method 0100) and analysis is
listed in Table 3.
9.3 Method 8315, Procedure 1, was tested by 12 laboratories using reagent
water and ground waters spiked at six concentrations over the range 30-2200
p.g/1. Method accuracy and precision were found to be directly related to the
concentration of the analyte and independent of the sample matrix. Mean recovery
weighted linear regression equations, calculated as a function of spike
concentration, as well as overall and single-analyst precision regression
equations, calculated as functions of mean recovery, are presented in Table 5.
These equations can be used to estimate mean recovery and precision at any
concentration value within the range tested.
8315A - 19 Revision 1
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10.0 REFERENCES
1. "OSHA Safety and Health Standards, General Industry", (29CRF1910).
Occupational Safety and Health Administration, OSHA 2206, (Revised, January
1976).
11.0 SAFETY
11.1 The toxicity or carcinogenicity of each reagent used in this method
has not been precisely defined; however, each chemical compound should be treated
as a potential health hazard. From this viewpoint, exposure to these chemicals
must be reduced to the lowest possible level by whatever means available. The
laboratory is responsible for maintaining a current awareness file of OSHA
regulations regarding the safe handling of the chemicals specified in this
method. A reference file of material safety data sheets should also be made
available to all personnel involved in the chemical analysis. Additional
references to laboratory safety are available.
11.2 Formaldehyde has been tentatively classified as a known or suspected,
human or mammalian carcinogen.
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TABLE 1
PROCEDURE 1 - METHOD DETECTION LIMITS" USING
LIQUID-SOLID EXTRACTION
Retention Time MDL
Analyte (minutes) (/ig/L)"
Formaldehyde 5.3 6.2
Acetaldehyde 7.4 43.7b
Propanal 11.7 11.0
Crotonaldehyde 16.1 5.9
Butanal 18.1 6.3
Cyclohexanone 27.6 5.8
Pentanal 28.4 15.3
Hexanal 34.1 10.7
Heptanal 35.0 10.0
Octanal 40.1 6.9
Nonanal 40.4 13.6
Decanal 44.1 4.4
" The method detection limit (MDL) is defined in Chapter One. With the exception
of acetaldehyde, all reported MDLs are based upon analyses of 6 to 8 replicate
blanks spiked at 25 M9/L-
b The reported MDL is based upon analyses of three replicate blanks fortified at
250 M9/L.
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TABLE 2
PROCEDURE 1 - METHOD DETECTION LIMITS8 USING
LIQUID-LIQUID EXTRACTION
Retention Time MDL
Analyte (minutes)
Formaldehyde
Acetaldehyde
Propanal
Crotonaldehyde
Butanal
Cyclohexanone
Pentanal
Hexanal
Heptanal
Octanal
Nonanal
Decanal
5.3
7.4
11.7
16.1
18.1
27.6
28.4
34.1
35.0
40.1
40.4
44.1
23.2
110.2"
8.4
5.9
7.8
6.9
13.4
12.4
6.6
9.9
7.4
13.1
The method detection limit (MDL) is defined in Chapter One. With the exception
of acetaldehyde, all reported MDLs are based upon analyses of 6 to 8 replicate
blanks spiked at 25
b The reported MDL is based upon analyses of three replicate blanks fortified at
250
8315A - 22 Revision 1
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TABLE 3
PROCEDURE 2 - SENSITIVITY (ppb, v/v) OF SAMPLING AND ANALYSIS FOR
CARBONYL COMPOUNDS IN AMBIENT AIR USING AN ADSORBENT CARTRIDGE
FOLLOWED BY GRADIENT HPLC"
Compound
10 20
30
Sample Volume (L)b
40 50 100 200
300 400 500
Acetaldehyde
Acetone
Acrolein
Benzaldehyde
Butyraldehyde
Crotonaldehyde
2,5-Dimethyl-
benzaldehyde
Formaldehyde
Hexanal
Isovaleraldehyde
Propionaldehyde
m-Tolualdehyde
o-Tolualdehyde
p-Tolualdehyde
Valeraldehyde
1.36
1.28
1.29
1.07
1.21
1.22
0.97
1.45
1.09
1.15
1.28
1.02
1.02
1.02
1.15
0.68
0.64
0.65
0.53
0.61
0.61
0.49
0.73
0.55
0.57
0.64
0.51
0.51
0.51
0.57
0.45
0.43
0.43
0.36
0.40
0.41
0.32
0.48
0.36
0.38
0.43
0.34
0.34
0.34
0.38
0.34
0.32
0.32
0.27
0.30
0.31
0.24
0.36
0.27
0.29
0.32
0.25
0.25
0.25
0.29
0.27
0.26
0.26
0.21
0.24
0.24
0.19
0.29
0.22
0.23
0.26
0.20
0.20
0.20
0.23
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
14
13
13
11
12
12
10
15
11
11
13
10
10
10
11
0.07
0.06
0.06
0.05
0.06
0.06
0.05
0.07
0.05
0.06
0.06
0.05
0.05
0.05
0.06
0.05
0.04
0.04
0.04
0.04
0.04
0.03
0.05
0.04
0.04
0.04
0.03
0.03
0.03
0.04
0.03
0.03
0.03
0.03
0.03
0.03
0.02
0.04
0.03
0.03
0.03
0.03
0.03
0.03
0.03
0.03
0.03
0.03
0.02
0.02
0.02
0.02
0.03
0.02
0.02
0.03
0.02
0.02
0.02
0.02
8 The ppb values are measured at 1 atm and 25°C. The sample cartridge is eluted
with 5 mL acetonitrile and 25 juL is injected into the HPLC. The maximum
sampling flow through a DNPH-coated Sep-PAK is about 1.5 L/minute.
b A sample volume of 1000 L was also analyzed.
of 0.01 ppb for all the target analytes.
The results show a sensitivity
8315A - 23
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TABLE 4
PERFORMANCE-BASED QC ACCEPTANCE LIMITS CALCULATED
USING THE COLLABORATIVE STUDY DATA
Spike
Analyte Concentration* Xb SRC
Formaldehyde
Propanal
Crotonaldehyde
Butanal
Cyclohexanone
Hexanal
Octanal
Decanal
160
160
160
160
160
160
160
160
154
148
160
151
169
151
145
153
30.5
22.4
34.8
22.7
39.2
34.6
40.1
40.0
Acceptance
Limits, %d
39
50
35
52
32
30
15
21
-153
-134
-165
-137
-179
-159
-166
-171
" Spike concentration, /xg/L.
b Mean recovery calculated using the reagent water, mean recovery, linear
regression equation, M9/L.
0 Overall standard deviation calculated using the reagent water, overall
standard deviation linear regression equation, p.g/1.
d Acceptance limits calculated as (X ± 3SR)100/spike concentration.
8315A - 24
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TABLE 5
WEIGHTED LINEAR REGRESSION EQUATIONS FOR MEAN RECOVERY AND PRECISION (/ig/L)
Applicable
Analyte Cone. Range
Formaldehyde 39.2 - 2450
Propanal 31.9 - 2000
Crotonaldehyde 32.4 - 2030
Butanal 35.4 - 2220
Cyclohexanone 31.6 - 1970
Hexanal 34.1- 2130
Octanal 32.9 - 2050
Decanal 33.2 - 2080
X
SR
sr
X
SR
sr
X
SR
X
SR
X
SR
sr
X
SR
Sr
X
SR
X
SR
Reagent Water
0.909C + 8.79
0.185X + 1.98"
0.093X + 5.79
0.858C + 10.49
0.140X + 1.63
0.056X + 2.76
0.975C + 4.36
0.185X + 5.15
0.096X + 1.85
0.902C + 6.65
0.149X + 0.21
0.086X - 0.71
0.962C + 14.97
0.204X + 4.73"
0.187X + 3.46
0.844C + 15.81
0.169X + 9.07
0.098X + 0.37"
0.856C + 7.88
0.200X + 11.17
0.092X + 1.71"
0.883C + 12.00
0.225X + 5.52
0.088X + 2.28"
Ground Water
0.870C + 14.84
0.177X + 13.85
0.108X + 6.24
0.892C + 22.22
0.180X + 12.37
0.146X + 2.08"
0.971C + 2.94
0.157X + 6.09
0.119X - 2.27
0.925C + 12.71
0.140X + 6.89
0.108X - 1.63"
0.946C + 28.95
0.345X + 5.02
0.123X + 7.64
0.926C + 9.16
0.132X + 8.31
0.074X - 0.40"
0.914C + 13.09
0.097X + 12.41
0.039X + 1.14
0.908C + 6.46
0.153X + 2.23
0.052X + 0.37
8 Variance is not constant over concentration range.
X Mean recovery, M9/L, exclusive of outliers.
SR Overall standard deviation, M9/U exclusive of outliers.
sr Single-analyst standard deviation, M9/U exclusive of outliers.
8315A - 25 Revision 1
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FIGURE la
PROCEDURE 2
LIQUID-SOLID PROCEDURAL STANDARD OF GROUP A ANALYTES AT 625
-0.80-
-1.00-
,-1.20-
o
v -». w
o <
-1.80-
-i.ao-
-a.oo-
i.oo
a.oo 3.00
x SO1 MinutM
4.00
Retention Time (min)
5.33
11.68
18.13
27.93
36.60
42.99
Analvte Derivative
Formaldehyde
Propanal
Butanal
Cyclohexanone
Heptanal
Nonanal
8315A - 26
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FIGURE Ib
PROCEDURE 1
LIQUID-SOLID PROCEDURAL STANDARD OF GROUP B ANALYTES AT 625
-0.60-
-0.80-
-1.00-
2-1.20H
-1.40-
-1.60-
-1.80-
1.00
a.oo 3.00
» 101 Minutes
4.00
Retention Time (min)
7.50
16.68
26.88
32.53
40.36
45.49
Analvte Derivative
Acetaldehyde
Crotonaldehyde
Pentanal
Hexanal
Octanal
Decanal
8315A - 27
Revision 1
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FIGURE 2a
PROCEDURE 1
LIQUID-LIQUID PROCEDURAL STANDARD OF GROUP A ANALYTES AT 625 /xg/L
-i.40
1.00
2.00 3.00
x iO1 ainutts
4.00
Retention Time (min)
5.82
13.23
20.83
29.95
37.77
43.80
Analvte Derivative
Formaldehyde
Propanal
Butanal
Cyclohexanone
Heptanal
Nonanal
8315A - 28
Revision 1
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FIGURE 2b
PROCEDURE 1
LIQUID-LIQUID PROCEDURAL STANDARD OF GROUP B ANALYTES AT 625 /jg/L
-8.00H
1.00
1.00
3.00
x 10* •inut««
4.00
Retention Time (min)
7.79
17.38
27.22
32.76
40.51
45.62
Analyte Derivative
Acetaldehyde
Crotonaldehyde
Pentanal
Hexanal
Octanal
Decanal
8315A - 29
Revision 1
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FIGURE 3
PROCEDURE 2
CHROMATOGRAPHIC SEPARATION OF THE DNPH DERIVATIVES OF 15 CARBONYL COMPOUNDS
DNPM
uuu
10
30
30
40
TIME. min
Peak Identification
Number
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
Compound
Concentration (nq/uL)
Formaldehyde 1.140
Acetaldehyde 1.000
Acrolein 1.000
Acetone 1.000
Propanal 1.000
Crotonaldehyde 1.000
Butanal 0.905
Benzaldehyde 1.000
Isovaleraldehyde 0.450
Pentanal 0.485
o-Tolualdehyde 0.515
m-Tolualdehyde 0.505
p-Tolualdehyde 0.510
Hexanal 1.000
2,4-Dimethylbenzaldehyde 0.510
8315A - 30
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METHOD 8315A
DETERMINATION OF CARBONYL COMPOUNDS
BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
Media (Optionl)
7.1.1 -7.1.1.1
Homogenize sample
and determne dry
weight
7.1.2 Extract
sample for 18
hours; fllter and
store extract
7 3.2 Measure 1-10
ml extract; adjust
volume to 100 mL
with water
7.0 What is
the sample
matrix?
7.0 Is media
solid or
aqueous''
Is sample
dear or sample
complexity
Known?
Stack Gas (Option
0
No
7.2.2 Centrifuge sample
at 2500 rpm for 10
; decant
and filter
Aqueous
7.3.1 Measure
aliquot of sample;
adjust volume to
100 ml with water
7.3.5.5 Exchange
solvent to methanol
0
8315A - 31
Revision 1
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METHOD 83ISA
continued
i
1
7.4.2.2 - 7.4.2.3
Connect sample cartridge
to dean syringe and
place in syringe rack
i
7.4.2.4 Backflush
cartridge with
acetonitrite
7.4.2.4
Doeseluaie
flow become
blocked?
7.4.24 Displace
trapped air with
acetonitrile in
syringe using a long-tip
disposable Pasteur pipet
7 4.2.5 Dilute to 5
mL with acetonitrile;
label flask; pipet 2
aliquots into
sample vials
7.4.2.6 Store
sample at 4C
8315A - 32
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METHOD 83ISA
continued
74.1.1 Measure volume
of aqueous phase of
sample; pour sample into
separator/ funnel and
drain methytene chloride
(from Method 0011) into
volumetric flask
7.4.1.2 Extract aqueous
solution with methytene
chloride; add methytene
chloride extracts to
volumetric flask
7.4.1.3 Dilute to volume
with metiytene chloride;
mix well; remove aliquot
7.4.1.5 Store
sample at 4C
7 4.1.4 Dilute
extract with mobile
phase
7.4.1.4 Exchange
solvent with methanol
as in 7.3.5.5
a high concentration
of formaldehyde?
7.4.1.4
Does sample
have a low
concentration of
formaldehyde?
7.4.1.4 Concentrate
extract during
solvent exchange
process
8315A - 33
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METHOD 83ISA
continued
7 5.2 Set LC conditions
to produce appropriate
retention times
75.1 Option
or2LC
conditions''
7 5.1 Set LC
conditions to produce
appropriate retention
times
7.5.2.1 Filter and
degas mobile phase
7.6.2 Process calibration
standards through same
processing steps as samples
7.6.3 - 7.6.4
Analyze solvent blank
and calibration standards;
tabulate peak areas
7.6.5 Determine response
factor at each concentration
76.5
Does
calibration
check meet
criteria?
O
7.6.5 Prepare new
calibration
standards
8315A - 34
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METHOD 83ISA
continued
7.6.6 - 7.6.7 Verity
calibration curve every day;
reanalyze 1 calibration
standard after 10
sample runs or less
7.7 Analyze samples
byHPLC
7.7.2 Inject a smaller
volume or dilute sample
7.7.4 Further
cleanup is required
7.7.2
Does peak
area exceed
calibration
curve'
7.7.4 Are
interferences
present?
7.8.1 Calculate each
response (actor, mean
response factor, and
percent RSD
i
1
7.8.2 - 7.8.5
Calculate analyte
concentrations
1
1
Stop
8315A - 35
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APPENDIX A
RECRYSTALLIZATION OF 2,4-DINITROPHENYLHYDRAZINE (DNPH)
NOTE: This procedure should be performed in a properly ventilated hood.
Inhalation of acetonitrile can result in nose and throat irritation
(brief exposure at 500 ppm) or more serious effects at higher
concentration and/or longer exposures.
A.I Prepare a saturated solution of DNPH by boiling excess DNPH in 200 ml
of acetonitrile for approximately 1 hour.
A.2 After 1 hour, remove and transfer the supernatant to a covered beaker
on a hot plate and allow gradual cooling to 40 to 60"C. Maintain this
temperature range until 95% of the solvent has evaporated, leaving crystals.
A.3 Decant the solution to waste and rinse the remaining crystals twice
with three times their apparent volume of acetonitrile.
A.4 Transfer the crystals to a clean beaker, add 200 ml of acetonitrile,
heat to boiling, and again let the crystals grow slowly at 40 to 60°C until 95%
of the solvent has evaporated. Repeat the rinsing process as in Sec. A.3.
A.5 Take an aliquot of the second rinse, dilute 10 times with
acetonitrile, acidify with 1 ml of 3.8 M perchloric acid per 100 ml of DNPH
solution, and analyze with HPLC as in Sec. 7.0 for Procedure 2. An acceptable
impurity level is less than 0.025 ng/juL of formaldehyde in recrystallized DNPH
reagent or below the sensitivity (ppb, v/v) level indicated in Table 3 for the
anticipated sample volume.
A.6 If the impurity level is not satisfactory, pipet off the solution to
waste, repeat the recrystallization as in Sec. A.4 but rinse with two 25 mL
portions of acetonitrile. Prep and analyze the second rinse as in Sec. A.5.
A.7 When the impurity level is satisfactory, place the crystals in an all-
glass reagent bottle, add another 25 ml of acetonitrile, stopper, and shake the
bottle. Use clean pipets when removing the saturated DNPH stock solution to
reduce the possibility of contamination of the solution. Maintain only a minimum
volume of the saturated solution adequate for day to day operation to minimize
waste of the purified reagent.
8315A - 36 Revision 1
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METHOD 8321A
SOLVENT EXTRACTABLE NON-VOLATILE COMPOUNDS BY
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY/THERMOSPRAY/MASS SPECTROMETRY
(HPLC/TS/MS) OR ULTRAVIOLET (UV) DETECTION
1.0 SCOPE AND APPLICATION
1.1 This method covers the use of high performance liquid chromatography
(HPLC), coupled with either thermospray-mass spectrometry (TS-MS), and/or
ultraviolet (UV), for the determination of disperse azo dyes, organophosphorus
compounds, and Tris-(2,3-dibromopropyl)phosphate, chlorinated phenoxyacid
compounds and their esters, and carbamates in wastewater, ground water, and
soil/sediment matrices. Data are also provided for chlorophenoxy acid herbicides
in fly ash (Table 15), however, recoveries for most compounds are very poor
indicating poor extraction efficiency for these analytes using the extraction
procedure included in this method. Additionally, it may apply to other
non-volatile compounds that are solvent extractable, are amenable to HPLC, and
are ionizable under thermospray introduction for mass spectrometric detection.
The following compounds can be determined by this method:
Compound Name
CAS No.'
Azo Dyes
Disperse Red 1
Disperse Red 5
Disperse Red 13
Disperse Yellow 5
Disperse Orange 3
Disperse Orange 30
Disperse Brown 1
Solvent Red 3
Solvent Red 23
Anthraquinone Dyes
Disperse Blue 3
Disperse Blue 14
Disperse Red 60
Coumarin Dyes
Fluorescent Briqhteners
Fluorescent Brightener 61
Fluorescent Brightener 236
Alkaloids
Caffeine
Strychnine
2872-52-8
3769-57-1
126038-78-6
6439-53-8
730-40-5
5261-31-4
17464-91-4
6535-42-8
85-86-9
2475-46-9
2475-44-7
17418-58-5
8066-05-5
3333-62-8
58-08-2
57-24-9
8321A - 1
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Compound Name
CAS No."
Organophosphorus Compounds
Methomyl 16752-77-5
Thiofanox 39196-18-4
Famphur 52-85-7
Asulam 3337-71-1
Dichlorvos 62-73-7
Dimethoate 60-51-5
Disulfoton 298-04-4
Fensulfothion 115-90-2
Merphos 150-50-5
Methyl parathion 298-00-0
Monocrotophos 919-44-8
Naled 300-76-5
Phorate 298-02-2
Trichlorfon 52-68-6
Tris-(2,3-Dibromopropy1) phosphate, (Tris-BP) 126-72-7
Chlorinated Phenoxyacid Compounds
Dalapon 75-99-0
Dicamba 1918-00-9
2,4-D 94-75-7
MCPA 94-74-6
MCPP 7085-19-0
Dichlorprop 120-36-5
2,4,5-T 93-76-5
Silvex (2,4,5-TP) 93-72-1
Dinoseb 88-85-7
2,4-DB 94-82-6
2,4-D, butoxyethanol ester 1929-73-3
2,4-D, ethylhexyl ester 1928-43-4
2,4,5-T, butyl ester 93-79-8
2,4,5-T, butoxyethanol ester 2545-59-7
Carbamates
Aldicarb* 116-06-3
Adicarb Sulfone 1646-88-4
Aldicarb Sulfoxide 1646-87-3
Aminocarb 2032-59-9
Barban 101-27-9
Benomyl 17804-35-2
Bromacil 314-40-9
Bendiocarb* 22781-23-3
Carbaryl* 63-25-2
Carbendazim* 10605-21-7
3-Hydroxy-Carbofuran 16655-82-6
Carbofuran* 1563-66-2
8321A - 2
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Compound Name CAS No."
Carbamates (continued)
Chloroxuron 1982-47-4
Chloropropham 101-21-3
Diuron 330-54-1
Fenuron 101-42-8
Fluometuron 2164-17-2
Linuron* 330-55-2
Methiocarb 2032-65-7
Methomyl* 16752-77-5
Mexacarbate 315-18-4
Monuron 150-68-5
Neburon 555-37-3
Oxamyl* 23135-22-0
Propachlor 1918-16-7
Propham 122-42-9
Propoxur 114-26-1
Siduron 1982-49-6
Tebuthiuron 34014-18-1
" Chemical Abstract Services Registry Number.
These carbamates were tested in a multi-laboratory evaluation; all
others were tested in a single-laboratory evaluation.
1.2 This method may be applicable to the analysis of other non-volatile
or semi volatile compounds.
1.3 Tris-BP has been classified as a carcinogen. Purified standard
material and stock standard solutions should be handled in a hood.
1.4 Method 8321 is designed to detect the chlorinated phenoxyacid
compounds (free acid form) and their esters without the use of hydrolysis and
esterification in the extraction procedure.
1.5 The compounds were chosen for analysis by HPLC/MS because they have
been designated as problem compounds that are hard to analyze by traditional
chromatographic methods (e.g. gas chromatography). The sensitivity of this
method is dependent upon the level of interferants within a given matrix, and
varies with compound class and even with compounds within that class.
Additionally, the limit of detection (LOD) is dependent upon the mode of
operation of the mass spectrometer. For example, the LOD for caffeine in the
selected reaction monitoring (SRM) mode is 45 pg of standard injected (10 pi
injection), while for Disperse Red 1 the LOD is 180 pg. The LOD for caffeine
under single quadrupole scanning is 84 pg and is 600 pg for Disperse Red 1 under
similar scanning conditions.
8321A - 3 Revision 1
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1.6 The experimentally determined limits of detection (LOD) for the
target analytes are presented in Tables 3, 10, 13, and 14. For further compound
identification, MS/MS (CAD - Collision Activated Dissociation) can be used as an
optional extension of this method.
1.7 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of high performance liquid chromatographs/mass
spectrometers and skilled in the interpretation of liquid chromatograms and mass
spectra. Each analyst must demonstrate the ability to generate acceptable
results with this method.
2.0 SUMMARY OF METHOD
2.1 This method provides reverse phase high performance liquid
chromatographic (RP/HPLC) and thermospray (TS) mass spectrometric (MS) conditions
for the detection of the target analytes. Quantitative analysis is performed by
TS/MS, using an external standard approach. Sample extracts can be analyzed by
direct injection into the thermospray or onto a liquid chromatographic-
thermospray interface. A gradient elution program is used on the chromatograph
to separate the compounds. Detection is achieved both by negative ionization
(discharge electrode) and positive ionization, with a single quadrupole mass
spectrometer. Since this method is based on an HPLC technique, the use of
ultraviolet (UV) detection is optional on routine samples.
2.2 Prior to the use of this method, appropriate sample preparation
techniques must be used.
2.2.1 Samples for analysis of chlorinated phenoxyacid compounds are
prepared by a modification of Method 8151 (see Sec. 7.1.2). In general,
one liter of aqueous sample or fifty grams of solid sample are pH
adjusted, extracted with diethyl ether, concentrated and solvent exchanged
to acetonitrile.
2.2.2 Samples for analysis of the other target analytes are prepared
by established extraction techniques. In general, water samples are
extracted at a neutral pH with methylene chloride, using an appropriate
3500 series method. An appropriate 3500 series method using methylene
chloride/acetone (1:1) is used for solid samples. A micro-extraction
technique is included for the extraction of Tris-BP from aqueous and
non-aqueous matrices.
2.2.3 For carbamates one liter aqueous samples or forty grams of
solid sample are methylene chloride extracted (refer to appropriate 3500
series method), concentrated (preferably using a rotary evaporator with
adapter) and solvent exchanged with methanol.
2.3 An optional thermospray-mass spectrometry/mass spectrometry
(TS-MS/MS) confirmatory method is provided. Confirmation is obtained by using
MS/MS Collision Activated Dissociation (CAD) or wire-repeller CAD.
8321A - 4 Revision 1
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3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, 8000 and 8151.
3.2 The use of Florisil Column Cleanup (Method 3620) has been
demonstrated to yield recoveries less than 85% for some of the compounds in this
method, and is therefore not recommended for all compounds. Refer to Table 2 of
Method 3620 for recoveries of organophosphorus compounds as a function of
Florisil fractions.
3.3 Compounds with high proton affinity may mask some of the target
analytes. Therefore, an HPLC must be used as a chromatographic separator, for
quantitative analysis.
3.4 Analytical difficulties encountered with specific organophosphorus
compounds, as applied in this method, may include (but are not limited to) the
following:
3.4.1 Methyl parathion shows some minor degradation upon analysis.
3.4.2 Naled can undergo debromination to form dichlorvos.
3.4.3 Merphos often contains contamination from merphos oxide.
Oxidation of merphos can occur during storage, and possibly upon
introduction into the mass spectrometer.
Refer to Method 8141 for other compound problems as related to the
various extraction methods.
3.5 The chlorinated phenoxy acid compounds, being strong organic acids,
react readily with alkaline substances and may be lost during analysis.
Therefore, glassware and glass wool must be acid-rinsed, and sodium sulfate must
be acidified with sulfuric acid prior to use to avoid this possibility.
3.6 Due to the reactivity of the chlorinated herbicides, the standards
must be prepared in acetonitrile. Methylation will occur slowly, if prepared in
methanol.
3.7 Benomyl is known to quickly degrade to carbendazim in the environment
(Reference 21).
3.8 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts or elevated baselines, or both, causing
misinterpretation of chromatograms or spectra. All of these materials must be
demonstrated to be free from interferences under the conditions of the analysis
by running reagent blanks. Specific selection of reagents and purification of
solvents by distillation in all-glass systems may be required.
3.9 Interferants co-extracted from the sample will vary considerably
from source to source. Retention times of target analytes must be verified by
using reference standards.
8321A - 5 Revision 1
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3.10 The optional use of HPLC/MS/MS methods aids in the confirmation of
specific analytes. These methods are less subject to chemical noise than other
mass spectrometric methods.
4.0 APPARATUS AND MATERIALS
4.1 HPLC/MS
4.1.1 High Performance Liquid Chromatograph (HPLC) - An analytical
system with programmable solvent delivery system and all required
accessories including 10-/zL injection loop, analytical columns, purging
gases, etc. The solvent delivery system must be capable, at a minimum, of
a binary solvent system. The chromatographic system must be capable of
interfacing with a Mass Spectrometer (MS).
4.1.1.1 HPLC Post-Column Addition Pump - A pump for post
column addition should be used. Ideally, this pump should be a
syringe pump, and does not have to be capable of solvent
programming.
4.1.1.2 Recommended HPLC Columns - A guard column and an
analytical column are required.
4.1.1.2.1 Guard Column - C18 reverse phase guard
column, 10 mm x 2.6 mm ID, 0.5 jiim frit, or equivalent.
4.1.1.2.2 Analytical Column - C18 reverse phase
column, 100 mm x 2 mm ID, 5 urn particle size of ODS-Hypersil;
or C8 reversed phase column, 100 mm x 2 mm ID, 3 /xm particle
size of MOS2-Hypersil, or equivalent.
4.1.2 HPLC/MS interface(s)
4.1.2.1 Micromixer - 10-jxL, interfaces HPLC column system
with HPLC post-column addition solvent system.
4.1.2.2 Interface - Thermospray ionization interface and
source that will give acceptable calibration response for each
analyte of interest at the concentration required. The source must
be capable of generating both positive and negative ions, and have
a discharge electrode or filament.
4.1.3 Mass spectrometer system - A single quadrupole mass
spectrometer capable of scanning from 1 to 1000 amu. The spectrometer
must also be capable of scanning from 150 to 450 amu in 1.5 sec. or less,
using 70 volts (nominal) electron energy in the positive or negative
electron impact modes. In addition, the mass spectrometer must be capable
of producing a calibrated mass spectrum for PEG 400, 600, or 800 (see Sec.
5.14).
8321A - 6 Revision 1
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4.1.3.1 Optional triple quadrupole mass spectrometer -
capable of generating daughter ion spectra with a collision gas in
the second quadrupole and operation in the single quadrupole mode.
4.1.4 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 any MS data file to be searched for ions of a specified mass, and
such ion abundances to be plotted 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 integration of the abundances in any
EICP between specified time or scan-number limits. There must be computer
software available to operate the specific modes of the mass spectrometer.
4.2 HPLC with UV detector - An analytical system with solvent
programmable pumping system for at least a binary solvent system, and all
required accessories including syringes, 10-juL injection loop, analytical
columns, purging gases, etc. An automatic injector is optional, but is useful
for multiple samples. The columns specified in Sec. 4.1.1.2 are also used with
this system.
4.2.1 If the UV detector is to be used in tandem with the
thermospray interface, then the detector cell must be capable of
withstanding high pressures (up to 6000 psi). However, the UV detector
may be attached to an HPLC independent of the HPLC/TS/MS and in that case
standard HPLC pressures are acceptable.
4.3 Purification Equipment for Azo Dye Standards
4.3.1 Soxhlet extraction apparatus.
4.3.2 Extraction thimbles, single thickness, 43 x 123 mm.
4.3.3 Filter paper, 9.0 cm (Whatman qualitative No. 1 or
equivalent).
4.3.4 Silica-gel column - 3 in. x 8 in., packed with Silica gel
(Type 60, EM reagent 70/230 mesh).
4.4 Extraction equipment for Chlorinated Phenoxyacid Compounds
4.4.1 Erlenmeyer flasks - 500-mL wide-mouth Pyrex®, 500-mL Pyrex®,
with 24/40 ground glass joint, 1000-mL Pyrex®.
4.4.2 Separatory funnel - 2000-mL.
4.4.3 Graduated cylinder - 1000-mL.
4.4.4 Funnel - 75 mm diameter.
4.4.5 Wrist shaker - Burrell Model 75 or equivalent.
4.4.6 pH meter.
8321A - 7 Revision 1
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4.5 Kuderna-Danish (K-D) apparatus (optional).
4.5.1 Concentrator tube - 10-mL graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.5.2 Evaporation flask - 500-mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.5.3 Snyder column - Two-ball micro (Kontes K-569001-0219 or
equivalent).
4.5.4 Springs - 1/2 in. (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.
4.5.5 Solvent vapor recovery system (Kontes K-545000-1006 or K-
547300-0000, Ace Glass 6614-30, or equivalent).
4.6 Disposable serological pipets - 5 ml x 1/10, 5.5 mm ID.
4.7 Collection tube - 15-mL conical, graduated (Kimble No. 45165 or
equivalent).
4.8 Vials - 5-mL conical, glass, with Teflon®-!ined screw-caps or crimp
tops.
4.9 Glass wool - Supelco No. 2-0411 or equivalent.
4.10 Microsyringes - 100-juL, 50-juL, 10-/A (Hamilton 701 N or equivalent),
and 50 nl (Blunted, Hamilton 705SNR or equivalent).
4.11 Rotary evaporator - Equipped with 1000-mL receiving flask.
4.12 Balances - Analytical, 0.0001 g, Top-loading, 0.01 g.
4.13 Volumetric flasks, Class A - 10-mL to 1000-mL.
4.14 Graduated cylinder - 100-mL.
4.15 Separatory funnel - 250-mL.
4.16 Separatory funnel - 2-liter, with Teflon® stopcock.
4.17 Concentrator adaptor (optional- for carbamate extraction).
8321A - 8 Revision 1
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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 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 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.
5.4 Ammonium acetate, NH4OOCCH3, solution (0.1 M). Filter through a 0.45
micron membrane filter (Millipore HA or equivalent).
5.5 Acetic acid, CH3C02H
5.6 Sulfuric acid solution
5.6.1 (1:1, v/v) - Slowly add 50 ml H2S04 (sp. gr. 1.84) to 50 ml
of water.
5.6.2 (1:3, v/v) - slowly add 25 ml H2S04 (sp. gr. 1.84) to 75 ml
of water.
5.7 Argon gas, 99+% pure.
5.8 Solvents
5.8.1 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.8.2 Toluene, C6H5CH3 - Pesticide quality or equivalent.
5.8.3 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.8.4 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 ethyl alcohol preservative must be
added to each liter of ether.
5.8.5 Methanol, CH3OH - HPLC quality or equivalent.
5.8.6 Acetonitrile, CH3CN - HPLC quality or equivalent.
5.8.7 Ethyl acetate CH3C02C2H5 - Pesticide quality or equivalent.
8321A - 9 Revision 1
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5.9 Standard Materials - pure standard materials or certified solutions
of each analyte targeted for analysis. Disperse azo dyes must be purified before
use according to Sec. 5.10.
5.10 Disperse Azo Dye Purification
5.10.1 Two procedures are involved. The first step is the Soxhlet
extraction of the dye for 24 hours with toluene and evaporation of the
liquid extract to dryness, using a rotary evaporator. The solid is then
recrystallized from toluene, and dried in an oven at approximately 100°C.
If this step does not give the required purity, column chromatography
should be employed. Load the solid onto a 3 x 8 inch silica gel column
(Sec. 4.3.4), and elute with diethyl ether. Separate impurities
chromatographically, and collect the major dye fraction.
5.11 Stock standard solutions - Can be prepared from pure standard
materials or can be purchased as certified solutions. Commercially-prepared
stock standards can be used if they are verified against EPA standards. If EPA
standards are not available for verification, then standards certified by the
manufacturer and verified against a standard made from pure material is
acceptable.
5.11.1 Prepare stock standard solutions by accurately weighing 0.0100
g of pure material. Dissolve the material in methanol or other suitable
solvent (e.g. prepare Tris-BP in ethyl acetate), and dilute to known volume
in a volumetric flask.
NOTE: Due to the reactivity of the chlorinated herbicides, the standards
must be prepared in acetonitrile. Methylation will occur if prepared
in methanol.
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.
5.11.2 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 frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards.
5.12 Calibration standards - A minimum of five concentrations for each
parameter of interest should be prepared through dilution of the stock standards
with methanol (or other suitable solvent). One of these concentrations should
be near, but above, the MDL. The remaining concentrations should correspond to
the expected range of concentrations found in real samples, or should define the
working range of the HPLC-UV/VIS or HPLC-TS/MS. Calibration standards must be
replaced after one or two months, or sooner if comparison with check standards
indicates a problem.
5.13 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system, along with the
8321A - 10 Revision 1
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effectiveness of the method in dealing with each sample matrix, by spiking each
sample, standard, and blank with one or two surrogates (e.g., organophosphorus
or chlorinated phenoxyacid compounds not expected to be present in the sample).
5.14 HPLC/MS tuning standard - Polyethylene glycol 400 (PEG-400), PEG-600
or PEG-800. Dilute to 10 percent (v/v) in methanol. Dependent upon analyte
molecular weight range: m.w. < 500 amu, use PEG-400; m.w. > 500 amu, use PEG-600,
or PEG-800.
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 Sample preparation - Samples for analysis of disperse azo dyes and
organophosphorus compounds must be prepared by an appropriate 3500 series method
prior to HPLC/MS analysis.:
Samples for the analysis of Tris-(2,3-dibromopropyl)phosphate wastewater
must be prepared according to Sec. 7.1.1 prior to HPLC/MS analysis. Samples for
the analysis of chlorinated phenoxyacid compounds and their esters should be
prepared according to Sec. 7.1.2 prior to HPLC/MS analysis.
7.1.1 Microextraction for Tris-BP:
7.1.1.1 Solid Samples
7.1.1.1.1 Weigh a 1-gram portion of the sample
into a tared beaker. If the sample appears moist, add an
equivalent amount of anhydrous sodium sulfate and mix well.
Add 100 jxL of Tris-BP (approximate concentration 1000 mg/L)
to the sample selected for spiking; the amount added should
result in a final concentration of 100 ng/juL in the 1-mL
extract.
7.1.1.1.2 Remove the glass wool plug from a
disposable serological pipet. Insert a 1 cm plug of clean
silane treated glass wool to the bottom (narrow end) of the
pipet. Pack 2 cm of anhydrous sodium sulfate onto the top of
the glass wool. Wash pipet and contents with 3 - 5 mL of
methanol.
7.1.1.1.3 Pack the sample into the pipet prepared
according to Sec. 7.1.1.1.2. If packing material has dried,
wet with a few mL of methanol first, then pack sample into
the pipet.
7.1.1.1.4 Extract the sampl e wi th 3 mL of methanol
followed by 4 mL of 50% (v/v) methanol/methylene chloride
(rinse the sample beaker with each volume of extraction
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solvent prior to adding it to the pipet containing the
sample). Collect the extract in a 15-mL graduated glass
tube.
7.1.1.1.5 Evaporate the extract to 1 mL using the
nitrogen blowdown technique (Sec. 7.1.1.1.6). Record the
volume. It may not be possible to evaporate some sludge
samples to a reasonable concentration.
7.1.1.1.6 Nitrogen Blowdown Technique
7.1.1.1.6.1Place 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.
7.1.1.1.6.2The internal wall of the
tube must be rinsed down several times with
methylene chloride 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. 7.1.1.1.7.
7.1.1.1.7 Transfer the extract to a glass vial
with a Teflon®-!ined screw-cap or crimp-top and store
refrigerated at 4°C. Proceed with HPLC analysis.
7.1.1.1.8 Determination of percent dry weight -
In certain cases, sample results are desired based on a dry
weight basis. When such data are desired, or required, a
portion of sample for this determination should be weighed
out at the same time as the portion used for analytical
determination.
WARNING: The drying oven should be contained in a hood or vented. Significant
laboratory contamination may result from drying a heavily contaminated
hazardous waste sample.
7.1.1.1.9 Immediately after weighing the sample
for extraction, weigh 5-10 g of the sample into a tared
crucible. Determine the % dry weight of the sample by drying
overnight at 105°C. Allow to cool in a desiccator before
weighing:
% dry weight = q of dry sample x 100
g of sample
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7.1.1.2 Aqueous Samples
7.1.1.2.1 Using a 100-mL graduated cylinder,
measure 100 ml of sample and transfer it to a 250-mL
separatory funnel. Add 200 /xL of Tris-BP (approximate
concentration 1000 mg/L) to the sample selected for spiking;
the amount added should result in a final concentration of
200 ng//iL in the 1-mL extract.
7.1.1.2.2 Add 10 ml of methylene chloride to the
separatory funnel. Seal and shake the separatory funnel
three times, approximately 30 seconds each time, with
periodic venting to release excess pressure.
NOTE: Methylene chloride creates excessive pressure rapidly; therefore,
initial venting should be done immediately after the separatory funnel
has been sealed and shaken once. Methylene chloride is a suspected
carcinogen, use necessary safety precautions.
7.1.1.2.3 Allow the organic layer to separate from
the water 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 phase separation. See Section 7.5,
Method 3510.
7.1.1.2.4 Collect the extract in a 15-mL graduated
glass tube. Proceed as in Sec. 7.1.1.1.5.
7.1.2 Extraction for chlorinated phenoxyacid compounds - Preparation
of soil, sediment, and other solid samples must follow Method 8151, with
the exception of no hydrolysis or esterification. (However, if the analyst
desires to determine all of the phenoxyacid moieties as the acid,
hydrolysis may be performed.) Section 7.1.2.1 presents an outline of the
procedure with the appropriate changes necessary for determination by
Method 8321. Section 7.1.2.2 describes the extraction procedure for
aqueous samples.
7.1.2.1 Extraction of solid samples
7.1.2.1.1 Add 50 g of soil/sediment sample to a
500-mL, wide mouth Erlenmeyer. Add spiking solutions if
required, mix well and allow to stand for 15 minutes. Add 50
ml of organic-free reagent water and stir for 30 minutes.
Determine the pH of the sample with a glass electrode and pH
meter, while stirring. Adjust the pH to 2 with cold H2S04
(1:1) and monitor the pH for 15 minutes, with stirring. If
necessary, add additional H2S04 until the pH remains at 2.
7.1.2.1.2 Add 20 ml of acetone to the flask, and
mix the contents with the wrist shaker for 20 minutes. Add
80 ml of diethyl ether to the same flask, and shake again for
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20 minutes. Decant the extract and measure the volume of
solvent recovered.
7.1.2.1.3 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.
7.1.2.1.4 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 2000
ml separatory funnel containing 250 ml of 5% acidified sodium
sulfate. 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.
7.1.2.1.5 Check the pH of the extract. If it is
not at or below pH 2, add more concentrated HC1 until the
extract is 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 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 the 500 ml
Erlenmeyer flask.
7.1.2.1.6 Add 45 - 50 g acidified anhydrous sodium
sulfate to the combined ether extracts. Allow the extract to
remain in contact with the sodium sulfate for approximately
2 hours.
NOTE: The drying step is very critical. Any moisture remaining in the ether
will result in low recoveries. The amount of sodium sulfate used 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 overnight in contact with the sodium sulfate.
7.1.2.1.7 Transfer the ether extract, through a
funnel plugged with acid-washed glass wool, into a 500-mL K-D
flask equipped with a 10 ml concentrator tube. Use a glass
rod to crush caked sodium sulfate during the transfer. Rinse
the Erlenmeyer flask and column with 20-30 ml of diethyl
ether to complete the quantitative transfer. Reduce the
volume of the extract using the macro K-D technique (Sec.
7.1.2.1.8).
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7.1.2.1.8 Add one or two clean boiling chips to
the flask and attach a three ball macro-Snyder column.
Attach the solvent vapor recovery glassware (condenser and
collection device) (Sec. 4.5.5) to the Snyder column of the
K-D apparatus following manufacturer's instructions. Prewet
the Snyder column by adding about 1 ml of diethyl ether to
the top. Place the apparatus on a hot water bath (600-65°C)
so that the concentrator tube is partially immersed in the
hot water and the entire lower rounded surface of the flask
is bathed in vapor. Adjust the vertical position of the
apparatus and the water temperature, as required, to complete
the concentration in 15-20 minutes. At the proper rate of
distillation the balls of the column will actively chatter,
but the chambers will not flood. When the apparent volume of
liquid reaches 5 ml, remove the K-D apparatus from the water
bath and allow it to drain and cool for at least 10 minutes.
7.1.2.1.9 Exchange the solvent of the extract to
acetonitrile by quantitatively transferring the extract with
acetonitrile to a blow-down apparatus. Add a total of 5 ml
acetonitrile. Reduce the extract volume according to Sec.
7.1.1.1.6, and adjust the final volume to 1 ml.
7.1.2.2 Preparation of aqueous samples
7.1.2.2.1 Using a 1000-mL graduated cylinder,
measure 1 liter (nominal) of sample, record the sample volume
to the nearest 5 ml, and transfer it to a separatory funnel.
If high concentrations are anticipated, a smaller volume may
be used and then diluted with organic-free reagent water to
1 liter. Adjust the pH to less than 2 with sulfuric acid
(1:1).
7.1.2.2.2 Add 150 ml of diethyl ether to the
sample bottle, seal, and shake for 30 seconds to rinse the
walls. Transfer the solvent wash to the separatory funnel and
extract the sample by shaking the funnel for 2 minutes with
periodic venting to release excess pressure. Allow the
organic layer to separate from the water layer 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. Drain the aqueous phase into a 1000-mL Erlenmeyer
flask.
7.1.2.2.3 Repeat the extraction two more times
using 100 ml of diethyl ether each time. Combine the
extracts in a 500 ml Erlenmeyer flask. (Rinse the 1000-mL
flask with each additional aliquot of extracting solvent to
make a quantitative transfer.)
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7.1.2.2.4 Proceed to Sec. 7.1.2.1.6 (drying, K-D
concentration, solvent exchange, and final volume
adjustment).
7.1.3 Extraction for carbamates - Preparation of soil, sediment, and
other solid samples must follow an appropriate 3500 series method.
7.1.3.1 Forty gram quantities are extracted with methylene
chloride using an appropriate 3500 series method.
7.1.3.2 Concentration steps can be achieved using a rotary
evaporator or K-D, to 5-10 ml volumes.
7.1.3.3 Final concentration and solvent exchange to 1-mL
final volume of methanol, can be done preferably using an adaptor on
the rotary evaporator. If an adaptor is unavailable, the final
concentration can be achieved using a gentle stream of nitrogen, in
a fume hood.
7.1.4 Extraction for carbamates - Preparation of aqueous samples
must follow an appropriate 3500 series method.
7.1.4.1 One liter quantities are extracted with methylene
chloride using an appropriate 3500 series method.
7.1.4.2 Final concentration and exchange to methanol is the
same as applied in Sees. 7.1.3.2 and 7.1.3.3.
7.2 Prior to HPLC analysis, the extraction solvent must be exchanged to
methanol or acetonitrile (Sec. 7.1.2.1.9). The exchange is performed using the
K-D procedures listed in all of the extraction methods.
7.3 HPLC Chromatographic Conditions:
7.3.1 Analyte-specific chromatographic conditions are shown in Table
1. Chromatographic conditions which are not analyte-specific are as
follows:
Flow rate: 0.4 mL/min
Post-column mobile phase: 0.1 M ammonium acetate (1% methanol)
(0.1 M ammonium acetate for phenoxyacid
compounds)
Post-column flow rate: 0.8 mL/min
7.3.2 If there is a chromatographic problem from compound retention
when analyzing for disperse azo dyes, organophosphorus compounds, and
Tris-(2,3-dibromopropyl)phosphate, a 2% constant flow of methylene chloride
may be applied as needed. Methylene chloride/aqueous methanol solutions
must be used with caution as HPLC eluants. Acetic acid (1%), another
mobile phase modifier, can be used with compounds with acid functional
groups.
7.3.3 A total flow rate of 1.0 to 1.5 mL/min is necessary to
maintain thermospray ionization.
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7.3.4 Retention times for organophosphorus compounds on the
specified analytical column are presented in Table 9.
7.4 Recommended HPLC/Thermospray/MS operating conditions:
7.4.1 Positive lonization mode
Repeller (wire or plate, optional):170 to 250 v (sensitivity
optimized). See Figure 2 for schematic of source with wire
repeller.
Discharge electrode: off
Filament: on or off (optional, analyte dependent)
Mass range: 150 to 450 amu (analyte dependent,
expect 1 to 18 amu higher than
molecular weight of the compound).
Scan time: 1.50 sec/scan.
7.4.2 Negative lonization mode
Discharge electrode: on
Filament: off
Mass Range: 135 to 450 amu
Scan time: 1.50 sec/scan.
7.4.3 Thermospray temperatures:
Vaporizer control: 110°C to 130°C.
Vaporizer tip: 200eC to 215eC.
Jet: 210°C to 220°C.
Source block: 230°C to 265°C. (Some compounds may
degrade in the source block at higher
temperatures, operator should use
knowledge of chemical properties to
estimate proper source temperature).
7.4.4 Sample injection volume: 20 juL is necessary in order to
overfill the 10-jiL injection loop. If solids are present in the extract,
allow them to settle or centrifuge the extract and withdraw the injection
volume from the clear layer.
7.5 Calibration:
7.5.1 Thermospray/MS system - Must be hardware-tuned on quadrupole
1 (and quadrupole 3 for triple quadrupoles) for accurate mass assignment,
sensitivity, and resolution. This is accomplished using polyethylene
glycol (PEG) 400, 600, or 800 (see Sec. 5.14) which has average molecular
weights of 400, 600, and 800, respectively. A mixture of these PEGs can
be made such that it will approximate the expected working mass range for
the analyses. Use PEG 400 for analysis of chlorinated phenoxyacid
compounds. The PEG is introduced via the thermospray interface,
circumventing the HPLC.
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7.5.1.1 The mass calibration parameters are as follows:
for PEG 400 and 600 for PEG 800
Mass range: 15 to 765 amu Mass range: 15 to 900 amu
Scan time: 5.00 sec/scan Scan time: 5.00 sec/scan
Approximately 100 scans should be acquired, with 2 to 3
injections made. The scan with the best fit to the accurate mass
table (see Tables 7 and 8) should be used as the calibration table.
7.5.1.2 The low mass range from 15 to 100 amu is covered by
the ions from the ammonium acetate buffer used in the thermospray
process: NH4+ (18 amu), NH4+H20 (36), CH3OHNH + (50) (methanol), or
CH3CNNH4+ (59) (acetonitrile), and CH3COOHNH4 (78) (acetic acid).
The appearance of the m/z 50 or 59 ion depends upon the use of
methanol or acetonitrile as the organic modifier. The higher mass
range is covered by the ammonium ion adducts of the various ethylene
glycols (e.g. H(OCH2CH2)nOH where n=4, gives the H(OCH2CH2)4OHNH/ ion
at m/z 212).
7.5.2 Liquid Chromatograph
7.5.2.1 Prepare calibration standards as outlined in Section
5.12.
7.5.2.2 Choose the proper ionization conditions, as outlined
in Sec. 7.4. Inject each calibration standard onto the HPLC, using
the chromatographic conditions outlined in Table 1. Calculate the
area under the curve for the mass chromatogram of each quantitation
ion. For example, Table 9 lists the retention times and the major
ions (>5%) present in the positive ionization thermospray single
quadrupole spectra of the organophosphorus compounds. In most cases
the (M+H)+ and (M+NH4) + adduct ions are the only ions of significant
abundance. Plot these ions as area response versus the amount
injected. The points should fall on a straight line, with a
correlation coefficient of at least 0.99 (0.97 for chlorinated
phenoxyacid analytes).
7.5.2.3 If HPLC-UV detection is also being used, calibrate
the instrument by preparing calibration standards as outlined in Sec.
5.12, and injecting each calibration standard onto the HPLC using the
chromatographic conditions outlined in Table 1. Integrate the area
under the full chromatographic peak for each concentration.
Quantitation by HPLC-UV may be preferred if it is known that sample
interference and/or analyte coelution are not a problem.
7.5.2.4 For the methods specified in Sees. 7.5.2.2 and
7.5.2.3, the retention time of the chromatographic peak is an
important variable in analyte identification. Therefore, the ratio
of the retention time of the sample analyte to the standard analyte
should be 1.0 - 0.1.
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7.5.2.5 The concentration of the sample analyte will be
determined by using the calibration curves determined in Sees. 7.5.2.2
and 7.5.2.3. These calibration curves must be generated on the same
day as each sample is analyzed. At least duplicate determinations
should be made for each sample extract. Samples whose concentrations
exceed the standard calibration range should be diluted to fall within
the range.
7.5.2.6 Refer to Method 8000 for further information on
calculations.
7.5.2.7 Precision can also be calculated from the ratio of
response (area) to the amount injected; this is defined as the
calibration factor (CF) for each standard concentration. If the
percent relative standard deviation (%RSD) of the CF is less than 20
percent 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. The CF and %RSD can be calculated as follows:
CF = Total Area of Peak/Mass injected (ng)
%RSD = SD/CF x 100
where:
SD = Standard deviation between CFs
CF = Average CF
7.6 Sample Analysis
7.6.1 Once the LC/MS system has been calibrated as outlined in Sec.
7.5, then it is ready for sample analysis. It is recommended that the
samples be initially analyzed in the negative ionization mode. If low
levels of compounds are suspected then the samples should also be screened
in the positive ionization mode.
7.6.1.1 A blank 20-jiL injection (methanol) must be analyzed
after the standard(s) analyses, in order to determine any residual
contamination of the Thermospray/HPLC/MS system.
7.6.1.2 Take a 20 juL aliquot of the sample extract from Sec.
7.4.4. Start the HPLC gradient elution, load and inject the sample
aliquot, and start the mass spectrometer data system analysis.
7.7 Calculations
7.7.1 Using the external standard calibration procedure (Method
8000), determine the identity and quantity of each component peak in the
sample reconstructed ion chromatogram which corresponds to the compounds
used for calibration processes. See Method 8000 for calculation equations.
7.7.2 The retention time of the chromatographic peak is an important
parameter for the identity of the analyte. However, because matrix
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interferences can change chromatographic column conditions, the retention
times are not as significant, and the mass spectra confirmations are
important criteria for analyte identification.
7.8 Optional Thermospray HPLC/MS/MS confirmation
7.8.1 With respect to this method, MS/MS shall be defined as the
daughter ion collision activated dissociation acquisition with quadrupole
one set on one mass (parent ion), quadrupole two pressurized with argon and
with a higher offset voltage than normal, and quadrupole three set to scan
desired mass range.
7.8.2 Since the thermospray process often generates only one or two
ions per compound, the use of MS/MS is a more specific mode of operation
yielding molecular structural information. In this mode, fast screening
of samples can be accomplished through direct injection of the sample into
the thermospray.
7.8.3 For MS/MS experiments, the first quadrupole should be set to
the protonated molecule or ammoniated adduct of the analyte of interest.
The third quadrupole should be set to scan from 30 amu to just above the
mass region of the protonated molecule.
7.8.4 The collision gas pressure (Ar) should be set at about 1.0
mTorr and the collision energy at 20 eV. If these parameters fail to give
considerable fragmentation, they may be raised above these settings to
create more and stronger collisions.
7.8.5 For analytical determinations, the base peak of the collision
spectrum shall be taken as the quantification ion. For extra specificity,
a second ion should be chosen as a backup quantification ion.
7.8.6 Generate a calibration curve as outlined in Sec. 7.5.2.
7.8.7 For analytical determinations, calibration blanks must be
analyzed in the MS/MS mode to determine specific ion interferences. If no
calibration blanks are available, chromatographic separation must be
performed to assure no interferences at specific masses. For fast
screening, the MS/MS spectra of the standard and the analyte could be
compared and the ratios of the three major (most intense) ions examined.
These ratios should be approximately the same unless there is an
interference. If an interference appears, chromatography must be utilized.
7.8.8 For unknown concentrations, the total area of the quantitation
ion(s) is calculated and the calibration curves generated as in Sec. 7.5
are used to attain an injected weight number.
7.8.9 MS/MS techniques can also be used to perform structural
analysis on ions represented by unassigned m/z ratios. The procedure for
compounds of unknown structures is to set up a CAD experiment on the ion
of interest. The spectrum generated from this experiment will reflect the
structure of the compound by its fragmentation pattern. A trained mass
spectroscopist and some history of the sample are usually needed to
interpret the spectrum. (CAD experiments on actual standards of the
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expected compound are necessary for confirmation or denial of that
substance.)
7.9 Optional wire-repeller CAD confirmation
7.9.1 See Figure 3 for the correct position of the wire-repeller in
the thermospray source block.
7.9.2 Once the wire-repeller is inserted into the thermospray flow,
the voltage can be increased to approximately 500 - 700 v. Enough voltage
is necessary to create fragment ions, but not so much that shorting occurs.
7.9.3 Continue as outlined in Section 7.6.
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 laboratory should also maintain records to document the quality of the data
generated.
8.2 Quality control procedures necessary to evaluate the HPLC system
operation are found in Method 8000, Sec. 7.0 and includes evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
Check the performance of the entire analytical system daily using data gathered
from analyses of blanks, standards, and replicate samples.
8.2.1 See Sec. 7.5.2.7 for required HPLC/MS parameters for standard
calibration curve %RSD limits.
8.2.2 See Sec. 7.5.2.4 regarding retention time window QC limits.
8.2.3 If any of the chromatographic QC limits are not met, the
analyst should examine the LC system for:
o Leaks,
o Proper pressure delivery,
o A dirty guard column; may need replacing or repacking, and
o Possible partial thermospray plugging.
Any of the above items will necessitate shutting down the HPLC/TS
system, making repairs and/or replacements, and then restarting the
analyses. The calibration standard should be reanalyzed before any sample
analyses, as described in Sec. 7.5.
8.2.4 The experience of the analyst performing liquid
chromatography is invaluable to the success of the method. Each day that
analysis is performed, the daily calibration standard should be evaluated
to determine if the chromatographic system is operating properly. If any
changes are made to the system (e.g. column change), the system must be
recalibrated.
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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, 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 Single operator accuracy and precision studies have been conducted
using spiked sediment, wastewater, sludge, and water samples. The results are
presented in Tables 4, 5, 6, 11, 12, 15, 20 and 21. Tables 4, 5, and 6 provide
single-laboratory data for Disperse Red 1, Table 11 with organophosphorus
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pesticides, Table 12 with Tris-BP, Table 15 with chlorophenoxyacid herbicides and
Tables 20 and 21 with carbamates.
9.2 LODs should be calculated for the known analytes, on each instrument
to be used. Tables 3, 10, and 13 list limits of detection (LOD) and/or estimated
quantitation limits (EQL) that are typical with this method.
9.2.1 The LODs presented in this method were calculated by analyzing
three replicates of four standard concentrations, with the lowest
concentration being near the instrument detection limit. A linear
regression was performed on the data set to calculate the slope and
intercept. Three times the standard deviation (3a) of the lowest standard
amount, along with the calculated slope and intercept, was used to find the
LOD. The LOD was not calculated using the specifications in Chapter One,
but according to the ACS guidelines specified in Reference 4.
9.2.2 Table 17 presents a comparison of the LODs from Method 8150
and Method 8321 for the chlorinated phenoxyacid compounds.
9.3 Table 16 presents multi-laboratory accuracy and precision data for the
chlorinated phenoxyacid herbicides. The data summary is based on data from three
laboratories that analyzed duplicate solvent solutions at each concentration
specified in the Table.
9.4 Tables 22 and 23 present the multi-laboratory accuracy and precision
data for the carbamates. The data summary is based on data from nine
laboratories that analyzed triplicate solvent solutions at each concentration
level specified in the Tables.
10.0 REFERENCES
1. Voyksner, R.D., Haney, C.A., "Optimization and Application of Thermospray
High-Performance Liquid Chromatography/Mass Spectrometry", Anal. Chem..
1985, 57, 991-996.
2. Blakley, C.R., Vestal, M.L., "Thermospray Interface for Liquid
Chromatography/Mass Spectrometry", Anal. Chem.. 1983, 5_5, 750-754.
3. Taylor, V., Hickey, D.M., Marsden, P.O., "Single Laboratory Validation of
EPA Method 8140", EPA-600/4-87/009, U.S. Environmental Protection Agency,
Las Vegas, NV, 1987, 144 pp.
4. "Guidelines for Data Acquisition and Data Quality Evaluation in
Environmental Chemistry", Anal. Chem., 1980, 52, 2242-2249.
5. Betowski, L.D., Jones, T.L., "The Analysis of Organophosphorus Pesticide
Samples by HPLC/MS and HPLC/MS/MS", Environmental Science and Technology,
1988.
8. U.S. EPA: 2nd Annual Report on Carcinogens, NTP 81-43, Dec. 1981, pp.
236-237.
9. Blum, A., Ames, B.N., Science 195, 1977, 17.
8321A - 23 Revision 1
January 1995
-------
10. Zweidinger, R.A., Cooper, S.D., Pellazari, E.D., Measurements of Organic
Pollutants in Water and Wastewater, ASTM 686.
11. Cremlyn, R., Pesticides: Preparation and mode of Action, John Wiley and
Sons, Chichester, 1978, p. 142.
12. Cotterill, E.G., Byast, T.H., "HPLC of Pesticide Residues in Environmental
Samples", In Liquid Chromatography in Environmental Analysis, Laurence,
J.F., Ed., Humana Press, Clifton, NJ, 1984.
13. Voyksner, R.D., "Thermospray HPLC/MS for Monitoring the Environment", In
Applications of New Mass Spectrometry Techniques in Pesticide Chemistry;
Rosen, J.D., Ed., John Wiley and Sons: New York, 1987.
14. Yinon, J., Jones, T.L., Betowski, L.D., Rap. Comm. Mass Spectrom., 1989,
3, 38.
15. Shore, F.L., Amick, E.N., Pan, S.T., Gurka, D.F., "Single Laboratory
Validation of EPA Method 8150 for the Analysis of Chlorinated Herbicides
in Hazardous Waste", EPA/600/4-85/060, U.S. Environmental Protection
Agency, Las Vegas, NV, 1985.
16. "Development and Evaluations of an LC/MS/MS Protocol", EPA/600/X-86/328,
Dec. 1986.
17. "An LC/MS Performance Evaluation Study of Organophosphorus Pesticides",
EPA/600/X-89/006, Jan. 1989.
18. "A Performance Evaluation Study of a Liquid Chromatography/Mass
Spectrometry Method for Tris-(2,3-Dibromopropyl) Phosphate",
EPA/600/X-89/135, June 1989.
19. "Liquid Chromatography/Mass Spectrometry Performance Evaluation of
Chlorinated Phenoxyacid Herbicides and Their Esters", EPA/600/X-89/176,
July 1989.
20. "An Inter!aboratory Comparison of an SW-846 Method for the Analysis of the
Chlorinated Phenoxyacid Herbicides by LC/MS", EPA/600/X-90/133, June 1990.
21. Somasundaram, L., and J.R. Coates, Ed., "Pesticide Transformation Products
Fate and Significance in the Environment", ACS Symposium Series 459,
Ch. 13, 1991.
22. Single-Laboratory Evaluation of Carbamates, APPL, Inc., Fresno, CA.
23. "Interlaboratory Calibration Study of a Thermospray-Liquid Chromatography/
Mass Spectrometry (TS-LC/MS) Method for Selected Carbamate Pesticides",
EPA/600/X-92/102, August 1992.
8321A - 24 Revision 1
January 1995
-------
TABLE 1
RECOMMENDED HPLC CHROMATOGRAPHIC CONDITIONS
Initial
Mobile Initial
Phase Time
(%) (min)
Analytes:
Orqanophosphorus Compounds
50/50 0
(water/methanol)
Azo Dyes (e.g. Disperse Red 1)
50/50 0
(water/CH3CN)
Tris-(2,3-dibromopropyl ) phosphate
50/50 0
(water/methanol)
Final
Gradient Mobile
(linear) Phase
(min) (%)
10 100
(methanol )
5 100
(CH3CN)
10 100
(methanol)
Final
Time
(min)
5
5
5
Chlorinated phenoxyacid compounds
75/25 2
(A/methanol)
40/60 3
(A/methanol)
15 40/60
(A/methanol)
75/25
10
(A/methanol)
Where A = 0.1 M ammonium acetate (1% acetic acid)
Carbamates
Option A:
Time
(mini
0
30
35
40
45
Mobile phase A
(percent)
95
20
0
95
95
Mobile phase B
(percent)
5
80
100
5
5
Where A = 5 mM ammonium acetate with 0.1 M acetic acid
and B = methanol
With optional post-column addition of 0.5 M ammonium acetate.
8321A - 25
Revision 1
January 1995
-------
Carbamates (continued)
Option B:
TABLE 1 (continued)
€
Mobile phase A Mobile phase B
(percent) (percent)
0 95 5
30 0 100
35 0 100
40 95 5
45 95 5
Where A = water with 0.1 M ammonium acetate with 1% acetic acid
B = methanol with 0.1 M ammonium acetate with 1% acetic acid
with optional post-column addition of 0.1 M ammonium acetate.
8321A - 26 Revision 1
January 1995
-------
TABLE 2
COMPOUNDS AMENABLE TO THERMOSPRAY MASS SPECTROMETRY
Disperse Azo Dyes Alkaloids
Methine Dyes Aromatic ureas
Arylmethane Dyes Amides
Coumarin Dyes Amines
Anthraquinone Dyes Amino acids
Xanthene Dyes Organophosphorus Compounds
Flame retardants Chlorinated Phenoxyacid Compounds
Carbamates
TABLE 3
LIMITS OF DETECTION AND METHOD SENSITIVITIES
FOR DISPERSE RED 1 AND CAFFEINE
Compound
Disperse Red 1
Caffeine
Mode
SRM
Single Quad
CAD
SRM
Single Quad
CAD
LOD
P9
180
600
2,000
45
84
240
EQL(7s)
P9
420
1400
4700
115
200
560
EQL(lOs)
pg
600
2000
6700
150
280
800
EQL = Estimated Quantitation Limit
Data from Reference 16.
8321A - 27 Revision 1
January 1995
-------
TABLE 4
PRECISION AND ACCURACY COMPARISONS OF MS AND MS/MS WITH
HPLC/UV FOR ORGANIC-FREE REAGENT WATER SPIKED WITH DISPERSE RED 1
Percent Recovery
Sample
HPLC/UV
MS
CAD
SRM
Spike 1
Spike 2
RPD
82.2 ± 0.2
87.4 + 0.6
6.1%
92.5 + 3.7
90.2 ± 4.7
2.5%
87.6 ± 4.6
90.4 ± 9.9
3.2%
95.5 + 17
90.0 ± 5.
5.9%
.1
9
Data from Reference 16.
TABLE 5
PRECISION AND ACCURACY COMPARISONS OF MS AND MS/MS WITH
HPLC/UV FOR MUNICIPAL WASTEWATER SPIKED WITH DISPERSE RED 1
Sample
Spike 1
Spike 2
RPD
HPLC/UV
93.4 ± 0
96.2 ± 0
3.0%
Percent Recovery
MS
.3 102.0 ± 31
.1 79.7 + 15
25%
CAD
82.7 + 13
83.7 ± 5.2
1.2%
Data from Reference 16.
8321A - 28
Revision 1
January 1995
-------
TABLE 6
RESULTS FROM ANALYSES OF ACTIVATED SLUDGE PROCESS WASTEWATER
Sample
5 mg/L Spiking
Concentration
1
1-D
2
3
RPD
0 mg/L Spiking
Concentration
1
1-D
2
3
RPD
Recovery
HPLC/UV
0.721 ± 0.003
0.731 ± 0.021
0.279 ± 0.000
0.482 ± 0.001
1.3%
0.000
0.000
0.000
0.000
--
of Disperse Red 1
MS
0.664 + 0.030
0.600 + 0.068
0.253 + 0.052
0.449 ± 0.016
10.1%
0.005 + 0.0007
0.006 ± 0.001
0.002 ± 0.0003
0.003 + 0.0004
18.2%
(mq/L)
CAD
0.796 + 0.008
0.768 + 0.093
0.301 + 0.042
0.510 + 0.091
3.6%
<0.001
<0.001
<0.001
<0.001
--
Data from Reference 16.
8321A - 29
Revision 1
January 1995
-------
TABLE 7
CALIBRATION MASSES AND % RELATIVE ABUNDANCES
OF PEG 400
Mass
18.0
35.06
36.04
50.06
77.04
168.12
212.14
256.17
300.20
344.22
388.25
432.28
476.30
520.33
564.35
608.38
652.41
653.41
696.43
697.44
% Relative
Abundances8
32.3
13.5
40.5
94.6
27.0
5.4
10.3
17.6
27.0
45.9
64.9
100
94.6
81.1
67.6
32.4
16.2
4.1
8.1
2.7
Intensity is normalized to mass 432.
8321A - 30 Revision 1
January 1995
-------
TABLE 8
CALIBRATION MASSES AND % RELATIVE ABUNDANCES
OF PEG 600
Mass
18.0
36.04
50.06
77.04
168.12
212.14
256.17
300.20
344.22
388.25
432.28
476.30
520.33
564.35
608.38
652.41
653.41
696.43
% Relative
Abundances"
4.7
11.4
64.9
17.5
9.3
43.9
56.1
22.8
28.1
38.6
54.4
64.9
86.0
100
63.2
17.5
5.6
1.8
Intensity is normalized to mass 564.
8321A - 31 Revision 1
January 1995
-------
TABLE 9
RETENTION TIMES AND THERMOSPRAY MASS SPECTRA
OF ORGANOPHOSPHORUS COMPOUNDS
Compound
Monocrotophos
Trichlorfon
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
Retention Time
(minutes)
1:09
1:22
1:28
4:40
9:16
9:52
10:52
13:30
13:55
18:51
Mass Spectra
(% Relative Abundance)"
241 (100), 224 (14)
274 (100), 257 (19), 238 (19)
230 (100), 247 (20)
238 (100), 221 (40)
398 (100), 381 (23), 238 (5),
221 (2)
326 (10), 309 (100)
281 (100), 264 (8), 251 (21),
234 (48)
278 (4), 261 (100)
292 (10), 275 (100)
315 (100), 299 (15)
a For molecules containing Cl, Br and S, only the base peak of the isotopic
cluster is listed.
Data from Reference 17.
8321A - 32
Revision 1
January 1995
-------
TABLE 10
PRECISION AND LIMITS OF DETECTION FOR
ORGANOPHOSPHORUS COMPOUND STANDARDS
Compound
Dichlorvos
Dimethoate
Phorate
Disulfoton
Fensulfothion
Naled
Merphos
Methyl
parathion
Ion
238
230
261
275
309
398
299
281
Standard
Quantitation
Concentration
(ng/ML)
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
%RSD
16
13
5.7
4.2
2.2
4.2
13
7.3
0.84
14
7.1
4.0
2.2
14
6.7
3.0
4.1
9.2
9.8
2.5
9.5
9.6
5.2
6.3
5.5
17
3.9
5.3
7.1
4.8
1.5
MDL (ng)
4
2
2
1
0.4
0.2
1
30
Data from Reference 17.
8321A - 33
Revision 1
January 1995
-------
TABLE 11
SINGLE OPERATOR ACCURACY AND PRECISION FOR LOW CONCENTRATION DRINKING
WATER (A), LOW CONCENTRATION SOIL (B), MEDIUM CONCENTRATION DRINKING
WATER (C), MEDIUM CONCENTRATION SEDIMENT (D)
Average
Recovery
Compound (%)
A
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
B
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
C
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
D
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
70
40
0.5
112
50
16
3.5
237
16
ND
ND
45
ND
78
36
118
52
146
4
65
85
10
2
101
74
166
ND
72
84
58
56
78
Standard
Deviation
7.7
12
1.0
3.3
28
35
8
25
4
5
15
7
19
4
29
3
7
24
15
1
13
8.5
25
8.6
9
6
5
4
Spike
Amount
uq/L
5
5
5
5
10
5
5
5
ttg/q
50
50
50
50
100
50
50
50
uq/L
50
50
50
50
100
50
50
50
mq/kq
2
2
2
2
3
2
2
2
Range of
Recovery
(%)
85 -
64 -
2 -
119 -
105 -
86 -
19 -
287 -
24 -
56 -
109 -
49 -
155 -
61 -
204 -
9 -
79 -
133 -
41 -
4 -
126 -
91 -
216 -
90 -
102 -
70 -
66 -
86 -
54
14
0
106
0
0
0
187
7
34
48
22
81
43
89
0
51
37
0
0
75
57
115
55
66
46
47
70
Number
of
Analyses
15
15
15
15
15
15
15
15
15
15
15
15
15
15
15
15
12
12
12
12
12
12
12
12
15
15
15
15
15
15
15
12
Data from Reference 17.
8321A - 34
Revision 1
January 1995
-------
TABLE 12
SINGLE OPERATOR ACCURACY AND PRECISION FOR MUNICIPAL WASTE
WATER (A), DRINKING WATER (B), CHEMICAL SLUDGE WASTE (C)
Compound
Tris-BP (A)
(B)
(C)
Average
Recovery
(%)
25
40
63
Spike
Standard Amount
Deviation (ng/juL)
8.0 2
5.0 2
11 100
Range
of %
Recovery
41 - 9.0
50 - 30
84 - 42
Number of
Analyses
15
12
8
Data from Reference 18.
Concentration
(ng/juL)
50
100
150
200
SINGLE
Average
Area
2675
5091
7674
8379
LOD
(ngM)
33
TABLE 13
OPERATOR EQL TABLE FOR
Standard 3*Std
Deviation Dev.
782 2347
558
2090
2030
Lower
EQL
(ng//*L)
113
TRIS-BP
7*Std
Dev.
5476
Upper
EQL
(ng/W
172
10*Std
Dev.
7823
EQL = Estimated Quantitation Limit
Data from Reference 18.
8321A - 35
Revision 1
January 1995
-------
TABLE 14
LIMITS OF DETECTION IN THE POSITIVE AND NEGATIVE ION MODES
FOR THE CHLORINATED PHENOXYACID HERBICIDES AND FOUR ESTERS
Compound
Dalapon
Dicamba
2,4-D
MCPA
Dichlorprop
MCPP
2,4,5-T
2,4,5-TP (Silvex)
Dinoseb
2,4-DB
2,4-D,Butoxy
ethanol ester
2,4,5-T,Butoxy
ethanol ester
2,4,5-T, Butyl
ester
2,4-D,ethyl-
hexyl ester
Positive Mode
Quantitation
Ion
Not detected
238 (M+NH4) +
238 (M+NH4)+
218 (M+NH4)+
252 (M+NH4)+
232 (M+NH4)+
272 (M+NH4)+
286 (M+NH4)+
228 (M+NH4-NO)+
266 (M+NH4) +
321 (M+Hr
372 (M+NH4) +
328 (M+NH4)+
350 (M+NH4)+
LOD
(ng)
13
2.9
120
2.7
5.0
170
160
24
3.4
1.4
0.6
8.6
1.2
Negative Mode
Quantitation
Ion
141 (M'H)'
184 (M'HCl)-
184 (M'HCl)-
199 (Ml)'
235 (Ml)'
213 (M'l)-
218 (M'HCl)-
269 (M'l)-
240 (M)'
247 (M'l)-
185 (MXeH^O,)-
195 (M-C8H1503)-
195 (M-CeH^O^-
161 (M-C10H1903)-
LOD
(ng)
11
3.0
50
28
25
12
6.5
43
19
110
Data from Reference 19.
8321A - 36
Revision 1
January 1995
-------
TABLE 15
SINGLE LABORATORY OPERATOR ACCURACY AND PRECISION
FOR THE CHLORINATED PHENOXYACID HERBICIDES
Compound
(a)
Average
Recovery(%)
Standard
Deviation
Spike
Amount
Range of
Recovery
Number
of
Analyses
LOW LEVEL DRINKING WATER
Dicamba
2,4-D
MCPA
MCPP
Dichlorprop
2,4,5-T
Silvex
2,4-DB
Dinoseb
Dalapon
2,4-D,ester
Dicamba
2,4-D
MCPA
MCPP
Dichlorprop
2,4,5-T
Silvex
2,4-DB
Dinoseb
Dalapon
2,4-D,ester
Dicamba
2,4-D
MCPA
MCPP
Dichlorprop
2,4,5-T
Silvex
2,4-DB
Dinoseb
Dalapon
2,4-D,ester
63
26
60
78
43
72
62
29
73
ND
73
HIGH LEVEL DRINKING
54
60
67
66
66
61
74
83
91
43
97
LOW LEVEL SAND
117
147
167
142
ND
134
121
199
76
ND
180
22
13
23
21
18
31
14
24
11
ND
17
WATER
30
35
41
33
33
23
35
25
10
9.6
19
26
23
79
39
ND
27
23
86
74
ND
58
5
5
5
5
5
5
5
5
5
5
5
50
50
50
50
50
50
50
50
50
50
50
M9/L
86 - 33
37 - 0
92 - 37
116 - 54
61 - 0
138 - 43
88 - 46
62 - 0
85 - 49
ND
104 - 48
M9/L
103
119
128
122
116
99
132
120
102
56
130
26
35
32
35
27
44
45
52
76
31
76
M9/9
147 - 82
180 -118
280 - 78
192 - 81
ND
171 - 99
154 - 85
245 - 0
210 - 6
ND
239 - 59
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
6
9
10
10
10
10
10
10
10
10
10
10
7
(a)All recoveries are in negative ionization mode, except for 2,4-D,ester.
ND = Not Detected.
8321A - 37
Revision 1
January 1995
-------
TABLE 15 (continued)
SINGLE LABORATORY OPERATOR ACCURACY AND PRECISION
FOR THE CHLORINATED PHENOXYACID HERBICIDES
Compound
(a)
Average
Recovery(%)
Standard
Deviation
Spike
Amount
Range of
Recovery
(%)
Number
of
Analyses
Dicamba
2,4-D
MCPA
MCPP
Dichlorprop
2,4,5-T
Silvex
2,4-DB
Dinoseb
Dalapon
2,4-D,ester
Dicamba
2,4-D
MCPA
MCPP
Dichlorprop
2,4,5-T
Silvex
2,4-DB
Dinoseb
Dalapon
2,4-D,ester
Dicamba
2,4-D
MCPA
MCPP
Dichlorprop
2,4,5-T
Silvex
2,4-DB
Dinoseb
Dalapon
2,4-D,ester
HIGH LEVEL SAND
153
218
143
158
92
160
176
145
114
287
20
LOW LEVEL MUNICIPAL
83
ND
ND
ND
ND
27
68
ND
44
ND
29
HIGH LEVEL MUNICIPAL
66
8.7
3.2
10
ND
2.9
6.0
ND
16
ND
1.9
33
27
30
34
37
29
34
22
28
86
3.6
ASH
22
ND
ND
ND
ND
25
38
ND
13
ND
23
ASH
21
4.8
4.8
4.3
ND
1.2
3.1
ND
6.8
ND
1.7
209 -119
276 -187
205 -111
226 -115
161 - 51
204 -131
225 -141
192 -110
140 - 65
418 -166
25 - 17
M9/9
104 - 48
ND
ND
ND
ND
60 - 0
128 - 22
ND
65 - 26
ND
53 - 0
M9/9
96
21
10
16
41
5
0
4.7
ND
3.6- 0
12 - 2.8
ND
23 - 0
ND
6.7- 0
9
9
9
9
9
9
9
9
9
9
7
9
9
9
9
9
9
9
9
9
9
6
9
9
9
9
9
9
9
9
9
9
6
''All recoveries are in negative ionization mode, except for 2,4-D,ester.
ND = Not Detected.
8321A - 38
Revision 1
January 1995
-------
TABLE 16
MULTI-LABORATORY ACCURACY AND PRECISION DATA
FOR THE CHLORINATED PHENOXYACID HERBICIDES
Compounds
Spiking
Concentration
Mean
% Relative
(% Recovery)" Standard Deviation
2,4,5-T
2,4,5-T,butoxy
2,4-D
2,4-DB
Dalapon
Dicamba
Dichlorprop
Dinoseb
MCPA
MCPP
Silvex
500 mq/L
90
90
86
95
83
77
84
78
89
86
96
23
29
17
22
13
25
20
15
11
12
27
50 mq/L
2,4,5-T 62
2,4,5-T,butoxy 85
2,4-D 64
2,4-DB 104
Dalapon 121
Dicamba 90
Dichlorprop 96
Dinoseb 86
MCPA 96
MCPP 76
Silvex 65
5 mq/L
2,4,5-T 90
2,4,5-T,butoxy 99
2,4-D 103
2,4-DB 96
Dalapon 150
Dicamba 105
Dichlorprop 102
Dinoseb 108
MCPA 94
MCPP 98
Silvex 87
" Mean of duplicate data from 3 laboratories.
b % RSD of duplicate data from 3 laboratories.
Data from Reference 20.
8321A - 39
68
9
80
28
99
23
15
57
20
74
71
28
17
31
21
4
12
22
30
18
15
15
Revision 1
January 1995
-------
TABLE 17
COMPARISON OF LODs: METHOD 8151 vs. METHOD 8321
lonization
Compound
Method 8151
Aqueous Samples
GC/ECD
EDLa (Mg/L)
Method 8321
Aqueous Samples
HPLC/MS/TS
LOD (Mg/L)
Mode
Dalapon
Dicamba
2,4-D
MCPA
Dichlorprop
MCPP
2,4,5-T
2,4,5-TP (Silvex)
2,4-DB
Dinoseb
1.3
0.081
0.2
0.056b
0".26
0.09
0.08
0.075
0.8
0.19
1.1
0.3
0.29
2.8
0.27
0.50
0.65
4.3
0.34
1.9
(-)
(-)
(+)
(-)
(+)
(+)
(-)
(-)
(+)
(-)
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.
40 CFR Part 136, Appendix B (49 FR 43234).
capillary column.
Chromatography using wide-bore
8321A - 40
Revision 1
January 1995
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TABLE 18
SINGLE-LABORATORY METHOD DETECTION LIMIT DETERMINATION
AND PRECISION RESULTS - WATERC
Analyte
Aldicarb sulfoxide"
Aldicarb sulfone
Oxamyl
Methomyl
3-Hydroxycarbofurana
Fenuron
Benomyl/Carbendazim
Aldicarb
Aminocarb
Carbofuran
Propoxur
Monuron
Bromacil
Tebuthiuron
Carbaryl
Fluometuron
Propham
Propachlor
Diuron
Siduron
Methiocarb
Barban
Linuron
Chloropropham
Mexacarbate
Chloroxuron
Neburon
Average %
Recovery
7.5
88.4
60.7
117
37.4
104
67.3
93.7
117
94.2
106
95.6
86.4
106
85.1
89.1
84.2
98.5
95.6
105
92.4
90.5
97.7
89.1
80.0
109
92.5
Standard
Deviation
0.27
0.44
0.10
0.49
0.25
0.20
0.13
0.46
0.53
0.17
0.32
0.24
0.12
0.17
0.29
0.19
0.15
0.16
0.14
0.27
0.16
0.79
0.19
0.68
1.41
0.32
0.14
%RSD
72.4
50.3
16.6
41.5
65.4
19.3
19.7
49.6
44.9
17.7
30.4
25.6
14.1
16.1
34.1
21.7
17.3
16.0
14.7
25.9
17.5
17.4
19.5
15.2
35.1
29.2
14.9
MDLb
M9/L
0.8
1.3
0.3
1.5
0.8
0.6
0.4
1.4
1.6
0.5
1.0
0.7
0.4
0.5
0.9
0.6
0.4
0.5
0.4
0.8
0.5
2.4
0.6
2.0
4.2
1.0
0.4
- Values generated from internal response factor calculations.
- Method detection limit determinations are based on twenty water
extractions. Aldicarb sulfoxide, Barban, Chloropropham, and Mexacarbate
spike levels were at 5 /ug/L. All other analytes were spiked at 1 M9/L.
The method detection limit was determined by multiplying the standard
deviation by 3. Quantitation was done using average linear regression
values, unless otherwise indicated.
- Data from Reference 22.
8321A - 41 Revision 1
January 1995
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TABLE 19
SINGLE-LABORATORY METHOD QUANTITATION LIMIT DETERMINATION
AND PRECISION RESULTS - SOILb
Analyte
Aldicarb sulfoxide
Aldicarb sulfone
Oxamyl
Methomyl
3-Hydroxycarbofuran
Fenuron
Benomyl/Carbendazim
Aldicarb
Aminocarb
Carbofuran
Propoxur
Monuron
Bromacil
Tebuthiuron
Carbaryl
Fluometuron
Propham
Propachlor
Diuron
Siduron
Methiocarb
Barban
Linuron
Chloropropham
Mexacarbate
Chloroxuron
Neburon
Average %
Recovery
66.9
118
89.6
86.8
103
91.2
68.0
72.0
84.4
102
95.2
107
99.6
96.8
99.6
92.8
100
114
101
107
124
108
113
104
62.2
97.6
110
Standard
Deviation
0.0492
0.0076
0.0049
0.0051
0.0116
0.0049
0.0082
0.0056
0.0082
0.0083
0.0091
0.0077
0.0069
0.0071
0.0054
0.0035
0.0039
0.0037
0.0060
0.0063
0.0054
0.0333
0.0037
0.0217
0.0119
0.0031
0.0044
%RSD
58.9
25.7
21.9
23.6
45.0
21.6
47.0
30.1
38.7
32.7
38.2
28.8
27.5
29.5
21.7
15.1
15.7
13.0
23.8
23.7
17.5
24.8
13.0
16.6
15.3
12.6
16.0
MDL"
M9/9
0.15
0.023
0.015
0.015
0.035
0.015
0.025
0.017
0.025
0.025
0.027
0.023
0.021
0.021
0.016
0.011
0.012
0.011
0.018
0.019
0.016
0.10
0.011
0.065
0.036
0.009
0.011
- Method detection limit determinations are based on twenty soil
extractions. Aldicarb sulfoxide, Barban, Chloropropham, and Mexacarbate
spike levels were at 0.125 M9/9- All other analytes were spiked at 0.025
M9/9- The method detection limit was determined by multiplying the
standard deviation by 3. Quantitation was done using average linear
regression values, unless otherwise indicated.
- Data from Reference 22.
8321A - 42 Revision 1
January 1995
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TABLE 20
SINGLE-LABORATORY EVALUATION OF AVERAGE RECOVERY
AND PRECISION DATA - WATER0
Analyte
Aldicarb sulfoxide
Aldicarb sulfone
Oxamyl8
Methomyl
3 -Hydroxycarbof uran"
Fenuron
Benomyl/Carbendazim
Aldicarb
Aminocarb
Carbofuran
Propoxur
Monuron
Bromacil
Tebuthiuron
Carbaryl
Fluometuron
Propham
Propachlor
Diuron
Siduron
Methiocarb
Barban
Linuron
Chloropropham
Mexacarbate
Chloroxuron
Neburon
Average %
Recovery6
7.6
56.0
38.9
52.0
22.2
72.5
47.3
81.0
109
85.5
79.1
91.8
87.6
87.1
82.1
84.4
80.7
84.3
90.8
88.0
93.3
88.1
87.1
94.9
79.8
106
85.3
Standard
Deviation
2.8
27.1
17.9
19.6
9.3
22.0
14.7
13.7
38.3
10.0
13.7
11.3
12.1
9.0
13.5
8.3
13.8
10.0
14.1
9.5
12.8
11.2
16.8
15.3
12.9
24.9
12.6
%RSD
37.0
48.5
45.9
37.7
41.7
30.3
31.0
16.9
35.1
11.7
17.3
12.3
13.8
10,3
16.5
9.8
17.1
11.9
15.6
10.8
13,8
12.7
19.3
16.1
16.2
23.5
14.8
- Values generated from internal response factor calculations.
- Nine spikes were performed at three concentrations. The concentrations
for Aldicarb sulfoxide, Barban, Chloropropham, and Mexacarbate spike
levels were at 25 /zg/L, 50 fj.g/1, and 100 /ig/L. All other analyte
concentrations were 5 M9/U 10 |ug/L, and 20 M9/L. One injection was
disregarded as an outlier. The total number of spikes analyzed was 26.
Quantitation was done using average linear regression values, unless
otherwise indicated.
- Data from Reference 22.
8321A - 43 Revision 1
January 1995
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TABLE 21
SINGLE-LABORATORY EVALUATION OF AVERAGE RECOVERY
AND PRECISION DATA - SOIL6
Analyte
Aldicarb sulfoxide
Aldicarb sulfone
Oxamyl
Methomyl
3-Hydroxycarbofuran
Fenuron
Benomyl/Carbendazim
Aldicarb
Aminocarb
Carbofuran
Propoxur
Monuron
Bromacil
Tebuthiuron
Carbaryl
Fluometuron
Propham
Propachlor
Diuron
Siduron
Methiocarb
Barban
Linuron
Chloropropham
Mexacarbate
Chloroxuron
Neburon
Average %
Recovery8
66.9
162
78.9
84.9
105
91.9
95.6
97.9
133
109
104
101
100
104
102
' 94.5
92.8
94.6
107
100
107
92.3
104
105
77.2
121
92.1
Standard
Deviation
31.3
51.4
46.1
25.8
36.3
16.7
18.2
17.0
44.7
14.4
16.5
12.4
9.0
11.9
15.5
15.7
12.0
10.3
17.4
12.0
14.2
15.6
13.6
9.3
9.8
27.3
16.5
%RSD
46.7
31.7
58.5
30.4
34.5
18.1
19.0
17.4
33.6
13.2
15.9
12.3
9.0
11.5
15.2
16.7
12.9
10.9
16.2
12.0
13.2
16.9
13.1
8.9
12.7
22.5
17.9
- Nine spikes were performed at three concentrations. The concentrations
for Aldicarb sulfoxide, Barban, Chloropropham, and Mexacarbate spike
levels were at 0.625 jug/g, 1.25 jug/g, and 2.5 M9/9- All other analyte
concentrations were 0.125 ng/g, 0.25 jug/9» and 0.50 jug/g. One injection
was disregarded as an outlier. The total number of spikes analyzed was
26. Quantitation was done using average linear regression values.
- Data from Reference 22.
8321A - 44
Revision 1
January 1995
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TABLE 22
MULTI-LABORATORY EVALUATION OF METHOD ACCURACY
(AFTER OUTLIER REMOVAL)"
Percent Recovery
Analyte
High-Concentration
Samples8
Medium-Concentration
Samplesb
Low-Concentration
Samples0
Aldicarb
Bendiocarb
Carbaryl
Carbendazim
Carbofuran
Diuron
Linuron
Methomyl
Oxamyl
98.7
81.4
92.0
125
87.8
79.9
84.8
93.3
83.8
110
95.0
108
138
92.3
98.8
93.0
90.8
88.0
52.0
52.0
62.0
128
72.0
66.0
82.0
90.0
98.0
- Three replicates per laboratory; eight to nine laboratories (per Table 26
of Reference 23). The true concentration is 90 mg/L per compound, except
Carbendazim at 22.5 mg/L.
- Two replicates per laboratory; eight to nine laboratories (per Table 26
of Reference 23). The true concentration is 40 mg/L per compound except
Carbendazim at 10 mg/L.
- Three replicates per laboratory; eight to nine laboratories (per Table 26
of Reference 23). The true concentration is 5 mg/L per compound, except
Carbendazim at 1.25 mg/L.
- Data from Reference 23.
8321A - 45
Revision 1
January 1995
-------
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FIGURE 1
SCHEMATIC OF THE THERMOSPRAY PROBE AND ION SOURCE
Flange
To
Trap
ft —
Mechanical
Pump
I
Source
Mounting
Plate
I
•lock
Ion Sampling
Cone
Ion*
electron Vaporiser
team
I
— LC
Vapor
Tomporaturt |
T4 Hock
Temperature
T.
Coupling
8321A - 47
Revision 1
January 1995
-------
FIGURE 2
THERMOSPRAY SOURCE WITH WIRE-REPELLER
(High sensitivity configuration)
i
CERAMIC INSULATOR
WIRE REPELLER
8321A - 48
Revision 1
January 1995
-------
FIGURE 3
THERMOSPRAY SOURCE WITH WIRE-REPELLER
(CAD configuration)
rxx
CERAMIC INSULATOR
WIRE REPELLER
8321A - 49
Revision 1
January 1995
-------
METHOD 8321
SOLVENT EXTRACTABLE NON-VOLATILE COMPOUNDS BY
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY/THERMQSPRAY/MASS SPECTROHETRY
(HPLC/TS/MS) OR ULTRAVIOLET (UV) DETECTION
l« aample
analyzed for
Trie-BP7
7.1 Prepare aample
for extreetion.
U
aample higK
concentration
wait*?
Uae dilution
Method 3580.
Chlorinated Phenoxyacid
Compound!
7.1.2 U«. modified
Method 8151.
Carbemate
Peeticidea
uee extraction
Method 3540 or
Method 3550.
Solid /aemple aolid
or equeoue?
Ua* •xtraction
Method 3610 or
Mothod 3620.
7.2 Exchange
• xtraetion •olvant
to mathanol or
aeatomtrila dunng
K-D procadurae.
7 3 3«t HPLC
Chroma tographie
condition*.
7.1.1 Prepare aample
for Tria-BP
microextrection.
7.1.1.1.1 Spike
the (elected (ample
wrthTne-BP.
71121 Spike
the aelected aample
with Tria-BP
7.1.1.1.1 Add
equivalent amount
of anhydroue
Na,SO.. .
i
7.1 1.2.2 Add
CH2CI2, •••)
•nd
thr«« t
7.1.1.1
eampla
1
7.1
.3 Pack
m pipat.
r
1.1.4
Extract aampla
firat with CH3OH
followed by
7 1.1.2.3
Allow organic
• nd water
l*y«r« to
7.1.1.1.6
Reduce volum*
fay K-0 or N 2
Slowdown.
7.1.1.2.4
Collect the
• xtrect
7.4 »«t HPLC/TS/MS
operating
condition*.
7.6 Instrument
Cehbretion
procedure.
7 fl Perform
HPLC/TS/MS
enelyeie.
7 7 U»e
Method 8000 to
calculate enalyta(e)
concentration.
8321A - 50
Revision 1
January 1995
-------
METHOD 8325
SOLVENT EXTRACTABLE NON-VOLATILE COMPOUNDS BY
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY/PARTICLE BEAM/MASS SPECTROMETRY
(HPLC/PB/MS)
1.0 SCOPE AND APPLICATION
1.1 This method describes the use of high performance liquid
chromatography (HPLC), coupled with particle beam (PB) mass spectrometry (MS),
for the determination of benzidines and nitrogen-containing pesticides in water
and wastewater. The following compounds can be determined by this method:
Compound CAS No.'
Benzidine 92-87-5
Benzoylprop ethyl 33878-50-1
Carbaryl 63-25-2
o-Chlorophenyl thiourea 5344-82-1
3,3'-Dichlorobenzidine 91-94-1
3,3'-Dimethoxybenzidine 119-90-4
3,3'-Dimethylbenzidine 612-82-8
Diuron 330-54-1
Linuron (Lorox) 330-55-2
Monuron 150-68-5
Rotenone 83-79-4
Siduron 1982-49-6
a Chemical Abstract Services Registry Number
1.2 The method also may be appropriate for the analysis of benzidines and
nitrogen-containing pesticides in non-aqueous matrices. The method may be
applicable to other compounds that can be extracted from a sample with methylene
chloride and are amenable to separation on a reverse phase liquid chromatography
column and transferable to the mass spectrometer with a particle beam interface.
1.3 Preliminary investigation indicates that the following compounds also
may be amenable to this method: Aldicarb sulfone, Carbofuran, Methiocarb,
Methomyl (Lannate), Mexacarbate (Zectran), and N-(l-Naphthyl)thiourea. Ethylene
thiourea and o-Chlorophenyl thiourea have been successfully analyzed by
HPLC/PB/MS, but have not been successfully extracted from a water matrix.
1.4 Tables 4-6 present method detection limits (MDLs) for the target
compounds, ranging from 2 to 25 M9/L. The MDLs are compound- and matrix-
dependent.
1.5 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of HPLC and skilled in the interpretation of
8325 - 1 Revision 0
January 1995
-------
particle beam mass spectrometry. Each analyst must demonstrate the ability to
generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 The target compounds for this method must be extracted from the sample
matrix prior to analysis.
2.1.1 Benzidines and nitrogen-containing pesticides are extracted
from aqueous matrices at a neutral pH with methylene chloride, using a
separatory funnel (Method 3510), a continuous liquid-liquid extractor
(Method 3520), or other suitable technique.
2.1.2 Solid samples are extracted using Methods 3540 (Soxhlet), 3541
(Automated Soxhlet), 3550 (Ultrasonic extraction), or other suitable
technique.
2.2 An aliquot of the sample extract is introduced into the HPLC
instrument and a gradient elution program is used to chromatographically separate
the target analytes, using reverse-phase liquid chromatography.
2.3 Once separated, the analytes are transferred to the mass spectrometer
via a particle beam HPLC/MS interface. Quantitation is performed using an
external standard approach.
2.4 An optional internal standard quantitation procedure is included for
samples which contain coeluting compounds or where matrix interferences preclude
the use of the external standard procedure.
2.5 The use of ultraviolet/visible (UV/VIS) detection is an appropriate
option for the analysis of routine samples, whose general composition has been
previously determined.
3.0 INTERFERENCES
3.1 Refer to Methods 3500 and 8000 for general discussions of
interferences with the sample extraction and chromatographic separation
procedures.
3.2 Although this method relies on mass spectrometric detection, which can
distinguish between chromatographically co-eluting compounds on the basis of
their masses, co-elution of two or more compounds will adversely affect method
performance. When two compounds coelute, the transport efficiency of both
compounds through the particle beam interface generally improves, and the ion
abundances observed in the mass spectrometer increase. The degree of signal
enhancement by coelution is compound-dependent.
3.2.1 This coelution effect invalidates the calibration curve and,
if not recognized, will result in incorrect quantitative measurements.
Procedures are given in this method to check for co-eluting compounds, and
must be followed to preclude inaccurate measurements.
8325 - 2 Revision 0
January 1995
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3.2.2 An optional internal standard calibration procedure has been
included for use in instances of severe co-elution or matrix interferences.
3.3 A major source of potential contamination is HPLC columns which may
contain silicon compounds and other contaminants that could prevent the
determination of method analytes. Generally, contaminants will be leached from
the columns into mobile phase and produce a variable background. Figure 1 shows
unacceptable background contamination from a column with stationary phase bleed.
3.4 Contamination may occur when a sample containing low analyte
concentrations is analyzed immediately after a sample containing relatively high
analyte concentrations. After analysis of a sample containing high analyte
concentrations, one or more method blanks should be analyzed. Normally, with
HPLC, this is not a problem unless the sample concentrations are at the percent
level.
4.0 APPARATUS AND MATERIALS
4.1 High performance liquid chromatograph (HPLC) - An analytical system
with programmable solvent delivery system and all necessary accessories including
5 nl injection loop, analytical columns, purging gases, etc. The solvent
delivery system must be capable, at a minimum, of handling a binary solvent
system, and must be able to accurately deliver flow rates between 0.20 - 0.40
mL/min. Pulse dampening is recommended, but not required. The chromatographic
system must be able to be interfaced with a mass spectrometer (MS). An
autoinjector is recommended and should be capable of accurately delivering 1 -
10 juL injections without affecting the chromatography.
4.1.1 HPLC Columns - An analytical column is needed, and a guard
column is highly recommended.
4.1.1.1 Analytical Column - Reverse phase column, C18
chemically bonded to 4-10 JLUH silica particles, 150 - 200 mm x 2 mm,
(Waters C-18 Novapak or equivalent). Residual acidic sites should be
blocked (endcapped) with methyl or other non-polar groups and the
stationary phase must be bonded to the solid support to minimize
column bleed. Select a column that exhibits minimal bleeding. New
columns must be conditioned overnight before use by pumping a 75 -
100% v/v acetonitrile:water solution through the column at a rate of
about 0.05 mL/min. Other packings and column sizes may be used if
appropriate performance can be achieved.
4.1.1.2 Guard Column - Packing similar to that used in
analytical column.
4.1.2 HPLC/MS interface - The particle beam HPLC/MS interface must
reduce the ion source pressure to a level compatible with the generation
of classical electron ionization (El) mass spectra, i.e., about 1 x 10"4 -
1 x 10"6 Torr, while delivering sufficient quantities of analytes to the
conventional El source to meet sensitivity, accuracy, and precision
requirements. The concentrations of background components with masses
greater than 62 Daltons should be reduced to levels that do not produce
8325 - 3 Revision 0
January 1995
-------
ions greater than a relative abundance of 10% in the mass spectra of the
analytes.
4.2 Mass spectrometer system - The mass spectrometer must be capable of
electron ionization at a nominal electron energy of 70 eV. The spectrometer
should be capable of scanning from 45 to 500 amu in 1.5 seconds or less
(including scan overhead). The spectrometer should produce a mass spectrum that
meets the criteria in Table 1 when 500 ng or less of DFTPPO are introduced into
the HPLC.
4.3 Data system - A computer system must be interfaced to the mass
spectrometer, and must be capable of the continuous acquisition and storage on
machine-readable media of all mass spectra obtained throughout the duration of
the chromatographic program. The computer software must be capable of searching
any HPLC/MS data file for ions of a specified mass and plotting such abundance
data versus time or scan number.
4.4 Volumetric flasks - Class A, in various sizes, for preparation of
standards.
4.5 Vials - 10-mL amber glass vials with Teflon®-lined screw caps or crimp
tops.
4.6 Analytical balance - capable of weighing 0.0001 g.
4.7 Extract filtration apparatus
4.7.1 Syringe - 10-mL, with Luer-Lok fitting.
4.7.2 Syringe filter assembly, disposable - 0.45 /^m pore size PTFE
filter in filter assembly with Luer-Lok fitting (Gelman Acrodisc, or
equivalent).
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 solvents must be HPLC-grade or equivalent.
5.3.1 Acetonitrile, CH3CN
5.3.2 Methanol, CH3OH
5.3.3 Ammonium acetate, NH4OOCCH3, (0.01M in water).
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5.4 Mobile phase - Two mobile phase solutions are needed, and are
designated Solvent A and Solvent B. Degas both solvents in an ultrasonic bath
under reduced pressure and maintain by purging with a low flow of helium.
5.4.1 Solvent A is a water:acetonitrile solution (75/25, v/v)
containing ammonium acetate at a concentration of 0.01M.
5.4.2 Solvent B is 100 % acetonitrile.
5.5 Stock standard solutions - Stock solutions may be prepared from pure
standard materials or purchased as certified solutions. Commercially-prepared
stock standards may be used at any concentration if they are certified by the
manufacturer.
5.5.1 Prepare stock standard solutions by accurately weighing 0.0100
g of pure material in a volumetric flask. Dilute to known volume in a
volumetric flask. If compound purity is certified at 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.
5.5.1.1 Dissolve benzidines and nitrogen-containing
pesticides in methanol, acetonitrile, or organic-free reagent water.
5.5.1.2 Certain analytes, such as 3,3'-dimethoxybenzidine,
may require dilution in 50% (v/v) acetonitrile:water or methanol :water
solution.
5.5.1.3 Benzidines may be used for calibration purposes in
the free base or acid chlorides forms. However, the concentration of
the standard should be calculated as the free base.
5.5.2 Transfer the stock standard solutions into amber bottles with
Teflon®-!ined screw-caps or crimp tops. 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.
5.6 Surrogate spiking solution - The recommended surrogates are
benzidine-D8, caffeine- N2, 3,3'-dichlorobenzidine-D6, andbis-(perfluorophenyl)-
phenylphosphine oxide. Prepare a solution of the surrogates in methanol or
acetonitrile at a concentration of 5 mg/mL of each. Other surrogates may be
included in this solution as needed. (A 10-^L aliquot of this solution added
to 1 L of water gives a concentration of 50 jiig/L of each surrogate). Store the
surrogate spiking solution in an amber vial in a freezer at -10°C or less.
5.7 MS performance check solution - Prepare a 100 ng//LtL solution of DFTPPO
in acetonitrile. Store this solution in an amber vial in a freezer at -10°C or
less.
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5.8 Calibration solutions
This method describes two types of calibration procedures that may be
applied to the target compounds: external standard calibration, and internal
standard calibration. Each procedure requires separate calibration standards.
In addition, the performance characteristics of the HPLC/PB/MS system indicate
that it may be necessary to employ a second order regression for calibration
purposes, unless a very narrow calibration range is chosen. See Method 8000 for
additional information on non-linear calibration techniques.
As described in Method 8000, the analyst has two options for performing a
non-linear calibration. The analyst may prepare five calibration standards and
perform triplicate analyses of each standard, or the analyst may prepare 10
calibration standards and perform a single analysis of each standard.
Whichever approach is used (a narrow linear range, the triplicate 5-point
approach, or the single 10-point approach), the lowest standard should be at a
concentration near, but above, the MDL of the target compound, and the remaining
standards should span the working range of the instrument.
5.8.1 For external standard calibration, prepare calibration
standards for all target compounds and surrogates in acetonitrile. DFTPPO
may be added to one or more calibration solutions to verify MS tune (see
Sec. 7.3). Store these solutions in amber vials at -10°C or less. Check
these solutions at least quarterly for signs of deterioration.
5.8.2 Internal standard calibration requires the use of suitable
internal standards (see Method 8000). Ideally, stable, isotopically-
labeled, analogs of the target compounds should be used. These labeled
compounds are included in the calibration standards and must also be added
to each sample extract immediately prior to analysis. Prepare the
calibration standards in a fashion similar to that for external standard
calibration, but include each internal standard in each of the calibration
standards.
The concentration of the internal standards should be 50 - 100 times
the lowest concentration of the unlabeled target compounds. In addition,
the concentration of the internal standards does not vary with the
concentrations of the target compounds, but is held constant. Store these
solutions in amber vials at -10°C or less. Check these solutions at least
quarterly for signs of deterioration.
5.9 Internal standard spiking solution - This solution is required when
internal standard quantitation is used. Prepare a solution containing each of
the internal standards that will be used for quantitation of target compounds
(see Sec. 5.8.2) in methanol. The concentration of this solution must be such
that a 1-juL volume of the spiking solution added to a 1-mL final extract will
result in a concentration of each internal standard that is equal to the
concentration of the internal standard in the calibration standards in Sec.
5.8.2. Store this solution in an amber vial at -10°C or less. Check this
solution at least quarterly for signs of deterioration. This solution is not
necessary if only external standard calibration will be used.
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5.10 Sodium chloride, NaCl - granular, used during sample extraction.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes, Sec.
4.1.
6.2 Samples should be extracted within 7 days and analyzed within 30 days
of extraction. Extracts should be stored in amber vials at -10°C or less.
7.0 PROCEDURE
7.1 Samples should be extracted by either Method 3510 (separatory funnel)
or Method 3520 (continuous extractor) or other appropriate technique. Prior to
extraction, add a 10-/uL aliquot of the surrogate spiking solution and 100 g of
sodium chloride to the sample, and adjust the pH of the sample to 7.0. Samples
of other matrices should be extracted by an appropriate sample preparation
technique. The concentration of surrogates in the sample should be 20-50 times
the method detection limit. Concentrate the extract to 1 mL, and exchange the
solvent to methanol, following the procedures in the extraction method.
7.2 Establish chromatographic, particle beam interface, and mass
spectrometer conditions, using the following conditions as guidance.
Mobile phase purge:
Mobile phase flow rate:
Gradient elution:
Desolvation chamber temperature:
Ion source temperature:
Electron energy:
Scan range:
Helium at 30 mL/min, continuous
0.25 - 0.3 mL/min through the column
Hold for 1 min at 25% acetonitrile
(Solvent A), then program linearly to
about 70% acetonitrile (60% Solvent
B) in 29 min. Start data acquisition
immediately.
45 - 80°C
250 - 290°C
70 eV
62 to 465 amu, at <1.5 sec/scan
NOTE: Post-column addition is an option that improves system precision and,
thereby, may improve sensitivity. Post-column flow rates depend on the
requirements of the interface and may range from 0.1 to 0.7 mL/min of
acetonitrile. Maintain a minimum of 30% acetonitrile in the interface.
Analyte-specific chromatographic conditions are also shown in Table 2.
(The particle beam interface conditions will depend on the type of nebulizer).
7.2.1 The analyst should follow the manufacturer's recommended
conditions for their interface's optimum performance. The interface is
usually optimized during initial installation by flow injection with
caffeine or benzidine, and should utilize a mobile phase of
acetonitrile/water (50/50, v/v). Major maintenance may require re-
optimization.
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7.2.2 Fine tune the interface by making a series of injections into
the HPLC column of a medium concentration calibration standard and
adjusting the operating conditions (Sec. 7.2) until optimum sensitivity and
precision are obtained for the maximum number of target compounds.
7.3 Initial calibration
7.3.1 Once the operating conditions have been established, calibrate
the MS mass and abundance scales using DFTPPO to meet the recommended
criteria in Table 1.
7.3.2 Inject a medium concentration standard containing DFTPPO, or
separately inject into the HPLC a 5-/iL aliquot of the 100 ng//uL DFTPPO
solution and acquire a mass spectrum. Use HPLC conditions that produce a
narrow (at least ten scans per peak) symmetrical peak. If the spectrum
does not meet the criteria (Table 1), the MS ion source must be retuned and
adjusted to meet all criteria before proceeding with calibration. An
average spectrum across the HPLC peak may be used to evaluate the
performance of the system.
Manual (not automated) ion source tuning procedures specified by the
manufacturer should be employed during tuning. Mass calibration should be
accomplished while an acetonitrile/water (50/50, v/v) mixture is pumped
through the HPLC column and the optimized particle beam interface. For
optimum long-term stability and precision, interface and ion source
parameters should be set near the center of a broad signal plateau rather
than at the peak of a sharp maximum (sharp maxima exhibit short-term
variations with particle beam interfaces and gradient elution conditions).
7.3.3 System performance criteria for the medium concentration
standard - Evaluate the stored HPLC/MS data with the data system software
and verify that the HPLC/PB/MS system meets the following performance
criteria.
7.3.3.1 HPLC performance - 3,3'-dimethylbenzidine and
3,3'-dimethoxybenzidine should be separated by a valley whose height
is less than 25% of the average peak height of these two compounds.
If the valley between them exceeds 25%, modify the gradient. If this
fails, the HPLC column requires maintenance. See Sec. 7.4.6.
7.3.3.2 Peak tailing - Examine a total ion chromatogram and
examine the degree of peak tailing. Severe tailing indicates a major
problem and system maintenance is required to correct the problem.
See Sec. 7.4.6
7.3.3.3 MS sensitivity - The signal-to-noise ratio for any
compound's spectrum should be at least 3:1.
7.3.3.4 Column bleed - Figure 1 shows an unacceptable
chromatogram with column bleed. Figure 2 shows an acceptable ion
chromatogram. Figure 3 is the mass spectrum of dimethyloctadecyl-
silanol, a common stationary phase bleed product. If unacceptable
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column bleed is present, the column must be changed or conditioned to
produce an acceptable background.
7.3.3.5 Coeluting compounds - Compounds which coelute cannot
be measured accurately because of carrier effects in the particle beam
interface. Peaks must be examined carefully for coeluting substances
and if coeluting compounds are present at greater than 10% of the
concentration of the target compound, either conditions must be
adjusted to resolve the components, or internal standard calibration
must be used.
7.3.4 Once optimized, the same instrument operating conditions must
be used for the analysis of all calibration standards, samples, blanks,
etc.
7.3.5 Once all the performance criteria are met, inject a 5-juL
aliquot of each of the other calibration standards using the same HPLC/MS
conditions.
7.3.5.1 The general method of calibration is a second order
regression of integrated ion abundances of the quantitation ions
(Table 3) as a function of amount injected. For second order
regression, a sufficient number of calibration points must be obtained
to accurately determine the equation of the curve. (See Method 8000
for the requirements for the number of standards that must be employed
for a non-linear calibration). Non-linear calibration models can be
applied to either the external standard or the internal standard
calibration approaches described here.
7.3.5.2 For some analytes the instrument response may be
linear over a narrow concentration range. In these instances, an
average calibration factor (external standard) or average response
factor (internal standard) may be employed for sample quantitation
(see Method 8000).
7.3.6 If a linear calibration model is used, calculate the mean
calibration factor or response factor for each analyte, including the
surrogates, as described in Method 8000. Calculate the standard deviation
(SD) and the relative standard deviation (RSD) as well. The RSD of an
analyte or surrogate must be less than or equal to 20%, if the linear model
is to be applied. Otherwise, proceed as described in Method 8000.
7.4 Calibration verification
Prior to sample analysis, verify the MS tune and initial calibration
at the beginning of each 8-hour analysis shift using the following
procedure:
7.4.1 Inject a 5-juL aliquot of the DFTPPO solution or a mid-level
calibration standard containing 500 ng of DFTPPO, and acquire a mass
spectrum that includes data for m/z 62-465. If the spectrum does not meet
the criteria in Table 1, the MS must be retuned to meet the criteria before
proceeding with the continuing calibration check.
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7.4.2 Inject a 5-/xL aliquot of a medium concentration calibration
solution and analyze with the same conditions used during the initial
calibration.
7.4.3 Demonstrate acceptable performance for the criteria shown in
Sec. 7.3.3.
7.4.4 Using the initial calibration (either linear or non-linear,
external standard or internal standard), calculate the concentrations in
the medium concentration calibration solution and compare the results to
the known values in the calibration solution. If calculated concentrations
deviate by more than 20% from known values, adjust the instrument and
inject the standard again. If the calibration cannot be verified with the
second injection, then a new initial calibration must be performed after
taking corrective actions such as those described in Sec. 7.9.
7.5 Sample Analysis
7.5.1 The column should be conditioned overnight before each use by
pumping a acetonitrile:water (70% v/v) solution through it at a rate of
about 0.05 mL/min.
7.5.2 Filter the extract through a 0.45 jum filter. If internal
standard calibration is employed, add 10 juL of the internal standard
spiking solution to the 1-mL final extract immediately before injection.
7.5.3 Analyze a 5-/iL aliquot of the extract, using the operating
conditions established in Sees. 7.2 and 7.3.
7.6 Qualitative identification
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 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 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.
7.6.2 The retention time of the sample component is within ± 10%
of the retention time of the standard.
7.6.3 The relative intensities of the characteristic ions agree
within 20% of the relative intensities of these ions in the reference
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spectrum. (Example: For an ion with an abundance of 50% in the reference
spectrum, the corresponding abundance in a sample spectrum can range
between 30% and 70%.)
7.6.4 Structural isomers that produce very similar mass spectra
should be identified as individual isomers if they have sufficiently
different HPLC 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.
7.6.5 Identification is hampered when sample components are not
resolved chromatographically and produce mass spectra containing ions
contributed by more than one analyte. When HPLC 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.
7.6.6 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.
7.7 Quantitative Analysis
7.7.1 Complete chromatographic resolution is necessary for accurate
and precise measurements of analyte concentrations. Compounds which
coelute cannot be measured accurately because of carrier effects in the
particle beam interface. Peaks must be examined carefully for coeluting
substances and if coeluting compounds are present at greater than 10% of
the concentration of the target compound, either conditions must be
adjusted to resolve the components, or the results for the target compound
must be flagged as potentially positively biased.
7.7.2 Calculate the concentration of each analyte, using either the
external standard or internal standard calibration. See Method 8000 for
the specific equations to be employed for either the non-linear or linear
calibration models.
7.7.3 If the response for any quantitation ion exceeds the initial
calibration range of the HPLC/PB/MS system, the sample extract must be
diluted and reanalyzed. When internal standard calibration is employed,
additional internal standard must be added to the diluted extract to
maintain the same concentration as in the calibration standards.
7.8 HPLC-UV/VIS Detection (optional)
7.8.1 Prepare calibration solutions as outlined in Sec. 5.8.
7.8.2 Inject 5 juL of each calibration solution onto the HPLC, using
the chromatographic conditions outlined in Sees. 7.2.1 and 7.2.2.
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Integrate the area under the full chromatographic peak at the optimum
wavelength (or at 230 nm if that option is not available) for each target
compound at each concentration.
7.8.3 The retention time of the chromatographic peak is an important
criterion for analyte identification. Therefore, the ratio of the
retention time of the sample analyte to the standard analyte should be 1.0
± 0.1.
7.8.4 Calculate calibration factors or response factors as described
in Method 8000, for either external standard or internal standard
calibration, and evaluate the calibration linearity as described in Method
8000.
7.8.5 Verify the calibration at the beginning of each 8-hour
analytical shift, as described above.
7.8.6 Once the calibration has been verified, inject a 5-/A aliquot
of the sample extract, start the HPLC gradient elution, load and inject the
sample aliquot, and begin data acquisition. Refer to Method 8000 for
guidance on calculation of concentration.
7.9 Corrective Actions
When the initial calibration cannot be verified, one or more of the
following corrective actions may be necessary.
7.9.1 Major maintenance such as cleaning an ion source, cleaning the
entrance lens, quadrapole rods, etc., will require a new initial
calibration.
7.9.2 Check and adjust HPLC and/or MS operating conditions; check
the MS resolution, and calibrate the mass scale.
7.9.3 Replace the mobile phases with fresh solvents. Verify that the
flow rate from the HPLC pump is constant.
7.9.4 Flush the HPLC column with acetonitrile.
7.9.5 Replace the HPLC column. This action will cause a change in
retention times.
7.9.6 Prepare fresh calibration solutions, and repeat the initial
calibration step.
7.9.7 Replace any components that leak.
7.9.8 Replace the MS electron multiplier, or any other faulty
components.
7.9.9 Clean the interface to eliminate plugged components and/or
replace skimmers according to the manufacturer's instructions.
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7.9.10 If peak areas are determined by the instrument software,
verify values by manual integration.
7.9.11 Increasing ion source temperature can reduce peak tailing, but
excessive ion source temperature can affect the quality of the spectra for
some compounds.
7.9.12 Air leaks into the interface may effect the quality of the
spectra (e.g. DFTPPO), especially when the ion source is operated at
temperatures in excess of 280°C.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control (QC)
procedures. Quailty control procedures to ensure the proper operation of the
various sample preparation 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 HPLC system
operation are found in Method 8000, Sec. 7.0 and includes evaluation of retention
time windows, calibration verification and chromatographic analysis of samples.
Necessary instrument QC is found in the following sections.
8.2.1 The HPLC/PB/MS system should be tuned to meet the DFTPPO
criteria in Sees. 7.3.1 and 7.4.1.
8.2.2 There should be an initial calibration of the HPLC/PB/MS
system as described in Sec. 7.3.
8.2.3 The HPLC/PB/MS system should meet the system performance
criteria in Sec. 7.3.3, each 8 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, 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.
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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 Single laboratory accuracy and precision data for the benzidines and
nitrogen-containing pesticides are presented in Tables 4-6. Five to seven 1-L
aliquots of organic-free reagent water, containing approximately five times the
MDL of each analyte, were analyzed with this procedure (Reference 1). The final
extract volume was 0.5 mL for these determinations.
9.1.1 Method detection limits (MDLs) are presented in Tables 4-6.
9.1.2 A multi-laboratory (12 laboratories) validation of the
determinative step was done for four of the analytes (benzidine, 3,3'-
dimethoxybenzidine, 3,3'-dimethylbenzidine, 3,3'-dichlorobenzidine).
Table 7 provides the results from this study for single laboratory
precision, overall laboratory precision, and overall laboratory accuracy.
The two concentration levels shown represent the two extremes of the
concentration range studied.
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10.0 REFERENCES
1. Bellar, T.A., Behymer, T.D., Ho, J.S., Budde, W.L., "Method 553:
Determination of Benzidines and Nitrogen-Containing Pesticides in Water by
Liquid-Liquid Extraction or Liquid-Solid Extraction and Reverse Phase High
Performance Liquid Chrotnatography/Particle Beam/Mass Spectrometry", U.S.
Environmental Protection Agency, EMSL-Cincinnati, Revision 1.1, August
1992.
2. Bellar, T.A., Behymer, T.D., Budde, W.L., "Investigation of Enhanced Ion
Abundances from a Carrier Process in High-Performance Liquid Chromatography
Particle Beam Mass Spectrometry", J. Am. Soc. Mass Spectrom., 1990, 1,
92-98.
3. Behymer, T.D., Bellar, T.A., and Budde, W.L., "Liquid
Chromatography/Particle Beam/Mass Spectrometry of Polar Compounds of
Environmental Interest", Anal. Chem., 1990, 62, 1686-1690.
4. Ho, J.S., Behymer, T.D., Budde, W.L., and Bellar, T.A., "Mass Transport and
Calibration in Liquid Chromatography/ Particle Beam/ Mass Spectrometry",
J. Am. Soc. Mass Spectrom., 1992, 3, 662-671.
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TABLE 1
ION ABUNDANCE CRITERIA FOR BIS(PERFLUOROPHENYL)PHENYLPHOSPHINE
(DECAFLUOROTRIPHENYLPHOSPHINE OXIDE, DFTPPO)
m/z
Relative Abundance
Purpose of Specification1
77
168
169
271
365
438
458
459
Present, major ion
Present, major ion
4 - 10% of 168
Present, major ion
5 - 10% of base peak
Present
Present
15 - 24% of mass 458
Low mass sensitivity
Mid-mass sensitivity
Mid-mass resolution and isotope ratio
Base peak
Baseline threshold check
Important high mass fragment
Molecular ion
High mass resolution and isotope
ratio
The primary use of all the ions is to check the mass calibration of the
mass spectrometer. The second use of these ions are the mass resolution
checks, including the natural isotope abundance ratios. The correct
setting of the baseline threshold is indicated by the presence of low
intensity ions, and is the third use of this test. Finally, the ion
abundance ranges may provide some standardization to fragmentation
patterns of the target compounds.
TABLE 2
RECOMMENDED HPLC CHROMATOGRAPHIC CONDITIONS
FOR BENZIDINES AND NITROGEN-CONTAINING PESTICIDES
Initial Mobile Initial
Phase (v/v %) Time (min)
Gradient
Time
Final Mobile
Phase (v/v %)
75/25
(water1/CH3CN)
29
30/70
(waterVCH3CN)
1 Water contains 0.01M ammonium acetate.
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TABLE 3
RETENTION TIME DATA AND QUANTITATION IONS FOR TARGET COMPOUNDS
Compound
Benzidine
Benzoylprop ethyl
Caffeine
Carbaryl
0-Chlorophenyl thiourea
3,3'-Dichlorobenzidine
3,3' -Dimethoxybenzi dine
3, 3' -Dimethyl benzi dine
Diuron
Ethylene thiourea
Linuron
Rotenone
Siduron
Retention
Time
System la
4.3
24.8
1.4
10.1
2.7
16.6
8.1
8.5
11.0
1.2
16.0
21.1
14.8
Retention
Time
System 2b
4.9
31.3
1.6
14.7
3.0
22.7
11.5
12.4
16.1
1.4
21.9
27.4
20.6
Quantitation
Ion
184
105
194
144
151
252
244
212
72
102
161
192
93
Surrogates:'
Benzidine-d8 4.2
Caffeine-15N2 1.3
3,3'-Dichlorobenzidine-d6 16.5
bi s(Perf1uorophenyl)-
phenylphosphine oxide 22.0
4.8
1.6
22.6
28.9
192
196
258
271
These retention times were obtained on a Hewlett-Packard 1090 liquid
chromatograph with a Waters C18 Novapak 15 cm x 2 mm column using gradient
conditions given in Table 1.
These retention times were obtained on a Waters 600 MS liquid
chromatograph with a Waters C18 Novapak 15 cm x 2 mm column using gradient
conditions given in Sec. 7.2.
These compounds cannot be used as surrogates if their unlabeled analogs
are present (see Sec. 3.2).
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TABLE 4
ACCURACY AND PRECISION DATA FROM SIX DETERMINATIONS OF THE TARGET COMPOUNDS
IN ORGANIC-FREE REAGENT WATER USING LIQUID-LIQUID EXTRACTION
Compound
Benzidine
Benzoylprop ethyl
Caffeine
Carbaryl
o-Chlorophenyl thiourea
3,3'-Dichlorobenzidine
3,3'-Dimethoxybenzidine
3,3'-Dimethylbenzidine
Diuron
Ethylene thiourea
Linuron
Monuron
Rotenone
Siduron
True
Cone.
(M9/L)
22.9
32.5
14.4
56.6
32.6
24.8
31.6
31.7
25.0
32.0
95.0
31.2
50.3
27.9
Mean
Observed
Cone.
(M9/L)
20.5
33.0
10.5
52.2
15.3
21.7
29.2
31.8
26.2
0.0
89.5
31.8
44.9
29.6
Std.
Dev.
(M9/L)
0.8
1.1
0.9
2.9
2.2
0.7
2.3
1.0
1.3
0.0
3.9
1.2
9.4
1.4
RSD
3.3
3.3
6.3
5.1
6.8
2.9
7.3
3.1
5.1
0.0
4.1
3.8
18.8
5.2
Mean
Accuracy
(% of
True)
89.6
101.6
72.6
92.3
47.0
89.6
92.3
100.4
104.8
0.0
94.2
101.9
89.3
106.3
MDL
(M9/L)
2.5
3.7
3.1
9.8
7.4*
2.4
7.7
3.3
4.4
*
13.1
4.0
31.6
4.7
* Not recovered
8325 - 18
Revision 0
January 1995
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TABLE 5
ACCURACY AND PRECISION DATA FROM SEVEN DETERMINATIONS OF THE TARGET COMPOUNDS
IN ORGANIC-FREE REAGENT WATER USING SOLID-PHASE EXTRACTION (C18 CARTRIDGE)8
Compound
Benzidine
Benzoylprop ethyl
Caffeine
Carbaryl
o-Chlorophenyl thiourea
3,3'-Dichlorobenzidine
3,3'-Dimethoxybenzidine
3,3' -Di methyl benzi dine
Diuron
Ethylene thiourea
Linuron
Monuron
Rotenone
Siduron
True
Cone.
(M9/L)
22.9
32.5
14.4
56.6
32.6
5.0
31.6
31.7
25.0
32.0
95.0
31.2
50.3
27.9
Mean
Observed
Cone.
(M9/L)
12.2
29.3
6.4
53.9
0.0
4.4
25.5
31.4
24.4
0.0
88.9
30.5
45.0
24.8
Std.
Dev.
(M9/L)
1.7
2.0
1.4
1.8
0.0
0.4
1.8
1.0
1.4
0.0
4.8
2.9
2.4
2.0
RSD
(%)
13.7
6.9
21.4
3.3
0.0
10.0
7.1
3.1
5.6
0.0
5.4
9.6
5.4
7.9
Mean
Accuracy
(% of
True)
53.2
90.2
44.2
95.2
0.0
89.6
80.8
99.0
97.6
0.0
93.6
97.8
89.6
88.9
MDL
(M9/L)
5.3
6.3
4.4
5.7
*
1.4
5.7
3.0
4.4
*
15.1
9.1
7.5
6.3
3 Reagent water contained 0.01 M ammonium acetate.
* Not recovered.
8325 - 19
Revision 0
January 1995
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TABLE 6
ACCURACY AND PRECISION DATA FROM SIX DETERMINATIONS OF THE TARGET COMPOUNDS
IN ORGANIC-FREE REAGENT WATER USING SOLID-PHASE EXTRACTION
(NEUTRAL POLYSTYRENE/DIVINYLBENZENE POLYMER DISK)
Compound
Benzidine
Benzoylprop ethyl
Caffeine
Carbaryl
o-Chlorophenyl thiourea
3,3'-Dichlorobenzidine
3,3'-Dimethoxybenzidine
3,3'-Dimethylbenzidine
Diuron
Ethylene thiourea
Linuron
Monuron
Rotenone
Siduron
True
Cone.
(M9/L)
22.9
32.5
14.4
56.6
32.6
5.0
31.6
31.7
25.0
32.0
95.0
31.2
50.3
27.9
Mean
Observed
Cone.
(M9/L)
24.7
31.1
0.7
59.5
0.0
5.0
32.8
31.5
26.1
0.0
97.9
34.4
40.5
26.8
Std.
Dev.
(M9/L)
2.4
3.0
0.5
4.7
0.0
0.5
2.2
2.1
1.8
0.0
8.7
2.5
6.0
1.0
RSD
(%)
9.8
9.6
72.5
7.9
0.0
9.4
6.7
6.7
7.0
0.0
9.0
7.3
14.8
3.6
Mean
Accuracy
(% of
True)
108.0
95.8
5.2
105.1
0.0
101.7
103.8
99.4
104.5
0.0
103.0
110.4
80.5
96.1
MDL
(M9/L)
8.1
10.1
1.8
15.8
*
1.6
7.4
7.1
6.1
*
29.3
8.4
20.2
3.4
* Not recovered.
8325 - 20
Revision 0
January 1995
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TABLE 7
MEAN RECOVERIES, MULTI-LABORATORY PRECISION AND ESTIMATES OF SINGLE ANALYST
PRECISION FOR THE MEASUREMENTS OF FOUR BENZIDINES BY LC/PB/MS
10 uq/L Test Cone. 100 uq/L Test Cone.
RSD RSD RSD RSD
Recovery Multi- Single Recovery Multi- Single
Compound (%) lab Analyst (%) lab Analyst
Benzidine 96 10 5.6 97 10 9.1
3,3'-Dimethoxybenzidine 104 20 18 95 10 7.0
3,3'-Dimethylbenzidine 98 14 10 97 8.6 4.9
3,3'-Dichlorobenzidine 96 18 9.4 97 9.1 4.6
8325 - 21 Revision 0
January 1995
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FIGURE 1
AN UNACCEPTABLE CHROMATOGRAM WITH COLUMN BLEED
COLUMN FOLLOWING EXPOSURE TO AMMONIUM ACETATE
80000-
48000-
40000-
38000-
30000-
23000-
coooo-
18000-
10000-
sooo-
0-
OIMETHYLOCTADECYL3ILANOL
(MAJOR IONS - M/Z 76, 313)
1C it 20 C4 C« 3C
100
90
-•0
79
-40
40
-40
30
•CO
10
-0
FIGURE 2
AN ACCEPTABLE CHROMATOGRAM FOLLOWING COLUMN FLUSHING
CIS COLUMN MAINTAINED WITH ACETONITRILE FLUSHING
24000-
20000-
16000-
12000-
4000-
4000-
0-
100
»0
•40
70
«0
-BO
-40
30
-CO
10
"k ic
CO 24 29 3C
8325 - 22
Revision 0
January 1995
-------
FIGURE 3
MASS SPECTRUM OF DIMETHYLOCTADECYL-SILANOL,
A COMMON STATIONARY PHASE BLEED PRODUCT
MASS SPECTRUM OF C18 COLUMN BLEED
100X 75
INT
I '
50
CH.
HO-Si-Ci
I
CHo
DIMETHYLOCTADECYLSILANOL
M-Methy
313
281
150
200
250
390
M/Z
8325 - 23
Revision 0
January 1995
-------
FIGURE 4
TOTAL ION CHROMATOGRAM OF ANALYTES AND SURROGATES
(140-950 ng Injected)
1200000
160 300 300 400 500 640
O.3ML/MIN O.O1 M AMMONIUM ACETATE
1100 1200 1300 1400
POST COLUMN 0.1 M L / M I N ACETONITRILE
8325 - 24
Revision 0
January 1995
-------
METHOD 8325
SOLVENT EXTRACTABLE NON-VOLATILE COMPOUNDS BY
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY/PARTICLE BEAM/MASS SPECTROMETRY
(HPLC/PB/MS)
7.1 Choose
appropriate extraction
technique.
7.6 Perform qualitative
identification of
compounds with
appropriate reference
spectrum.
7.2 Establish
chromatographic,
particle beam
interference, and mass
spectrometer
conditions.
7.7 Calculate
concentrations of
each sample.
Yes
7.3 Calibrate
equipment.
7.7.3 Dilute
sample.
7.4 Verify
calibration.
7.5 Perform
sample analysis.
7.7.3
Does
response exceed
the calibration
range?
7.8
HPLC:
UV/UIS
detection be
performed?
(Optional)
7.8.1 - 7.8.4
Calibrate equipment
7.8.5 Verify
calibration.
8325 - 25
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January 1995
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METHOD 8332
NITROGLYCERINE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8332 is used to determine the concentration of nitroglycerine
(CAS No. 55-63-0) in aqueous matrices such as waste water, groundwater, and
surface water. This method may also be applicable to other matrices. Method
8332 also provides a qualitative procedure for determining the presence or
absence of nitroglycerine (See Appendix A).
1.2 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of HPLC instrumentation and skilled in the
interpretation of HPLC chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 Samples are injected onto a reverse phase high performance liquid
chromatograph (HPLC) column and the nitroglycerine concentration is quantitated
using a UV detector.
3.0 INTERFERENCES
3.1 Solvents, reagents, glassware and other sample processing hardware
may yield discrete artifacts and/or elevated baselines, causing misinterpretation
of the chromatograms. All of these materials must be demonstrated to be free
from interferences.
4.0 APPARATUS AND MATERIALS
4.1 HPLC system
4.1.1 HPLC (isocratic) - equipped with a pump, a direct injection
port or 20 nl loop injector and a 214 nm UV detector.
4.1.2 Column - Waters Radial - Pak CN, 10 nm particle size (or
equivalent).
4.1.3 Integrator.
4.2 Injection syringe.
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
8332 - 1 Revision 0
January 1995
-------
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 Acetonitrile, CH3CN.
5.4 Nitroglycerine - "Nitrostat" tablets (nominal concentration =
0.4 mg/tablet), available from Warner Lambert/Parke Davis. Certificate of
Analysis for any current lot is available from Warner Lambert, Lititz, Pa.
(717) 626-2011.
5.5 HPLC mobile phase eluant - 60% Acetonitrile/40% Organic-free reagent
water.
5.6 HPLC calibration standards - Dissolve five nitroglycerine tablets in
the HPLC mobile phase, using a 500 mL volumetric flask. Dilute to volume with
mobile phase. The resulting standard will contain 4 mg/L nitroglycerine. 8 mg/L
and 12 mg/L nitroglycerine standard solutions can be prepared in the same manner.
Other concentrations, as needed, can be prepared by appropriate dilutions. Use
the concentration listed on the "Nitrostat" Certificate of Analysis to calculate
the concentration of the standards to three significant figures.
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 Chromatographic Conditions (Recommended):
Flow rate: 1.0 mL/min.
UV Detector: 214 nm.
Injection: 20 /zL (for 4 to 12 mg/L nitroglycerine).
Chromatographic conditions should be established to give a retention time
for nitroglycerine of approximately 5.1 min.
7.2 Initial Calibration - Prepare a 5-point calibration curve and
establish the calibration range of the method (nominally between 4 mg/L and
12 mg/L). Calculate a correlation coefficient, slope, and zero intercept from
the regression analysis of the data points (peak area vs. concentration).
7.3 Make 20 ^L injections for each sample or sample dilution. If the
response for the sample exceeds that of the initial calibration range, the sample
must be diluted. Calculate sample concentrations using the slope and the
intercept figures from the regression analysis of the standards.
8332 - 2 Revision 0
January 1995
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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 laboratory should also maintain records to document the quality of the data
generated.
8.2 Quality control procedures necessary to evaluate the HPLC 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.
8332 - 3 Revision 0
January 1995
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9.0 METHOD PERFORMANCE
9.1 Method 8332 was tested by one laboratory using two industrial
wastewater samples. The wastewaters were analyzed at two dilutions with three
injections of each dilution. The results of these analyses are shown in Table 1.
10.0 REFERENCES
1. U.S. Department of the Treasury; Bureau of Alcohol, Tobacco and Firearms;
Lab Number 88-N-0648 B.
8332 - 4 Revision 0
January 1995
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TABLE 1
SINGLE LABORATORY PRECISION
Nitroglycerine by Weight
As parts per Million As Percent
1036 ± 38 0.1036 ± 0.0038
952 ± 53 0.0952 ± 0.0053
The numbers shown are the average concentration plus or minus one standard
deviation calculated for six analyses on each sample.
8332 - 5 Revision 0
January 1995
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METHOD 8332
NITROGLYCERINE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY
7.1 Establish
appropriate
chromatographic
conditions.
7.2 Perform
calibration.
7.3
Does
sample response1
exceed
calibration
range?
7.3 Calculate sample
concentrations.
8332 - 6
Revision 0
January 1995
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APPENDIX A
THIN-LAYER CHROMATOGRAPHY PROCEDURE FOR QUALITATIVE DETERMINATION
OF THE PRESENCE OF NITROGLYCERINE
A. 1.0 APPARATUS
A. 1.1 Thin-layer chromatography (TLC) system
A.1.1.1 TLC plate, Analtech silica gel GHL (Catalog # 11511), or
equivalent.
A.1.1.2 Device (syringe, capillary pipet or other) for spotting
TLC plate.
A.1.1.3 TLC developing tank.
A.1.1.4 Sprayer.
A.2.0 REAGENTS
A.2.1 Solvents
A.2.1.1 Acetonitrile, CH3CN.
A.2.1.2 1,2-Dichloroethane, C1CH2CH2C1.
A.2.1.3 Carbon tetrachloride, CC14.
A.2.1.4 Ethanol (absolute), CH3CH2OH.
A.2.1.5 Acetone, CH3COCH3.
A.2.1.6 Tetrahydrofuran, C4H80.
A.2.2 Diphenylamine, (C6H5)2NH.
A.2.3 alpha-Naphthylamine, C10H7NH2.
A.2.4 Sulfanilic acid, 4-(H2N)C6H4S03H.
A.2.5 Sulfuric acid, H2S04.
A.2.6 Potassium Hydroxide, KOH.
A.2.7 Acetic acid (30%), CH3C02H.
A.2.8 TLC developing solvent - 20% dichloroethane/80% carbon
tetrachloride.
A.2.9 TLC overspray - Prepare a solution of 5% diphenylamine in ethanol.
8332-7 Revision 0
Appendix A January 1995
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For very weak spots, a solution of 5% diphenylamine in concentrated sulfuric acid
may be required.
A.2.10 Alternative TLC overspray (Greiss reagent) -
1. Alcoholic KOH (2%)
2. 1% sulfanilic acid in acetic acid (30%)
3. 1% alpha-naphthylamine in acetic acid (30%)
A.2.11 TLC standard - Use double-base smokeless powder (e.g. Hercules Red
Dot, Bullseye, or Winchester-Western ball powder) available at any gun shop.
Dissolve in acetone, acetonitrile, or THF.
A.3.0 PROCEDURE
A.3.1 Add a quantity of the developing solvent mixture to the developing
tank that will be sufficient to continuously wet the edge of the TCL plate.
Cover the tank tightly, and allow the vapor phase to equilibrate (several hours).
A.3.2 Streak or spot the sample near the edge of the plate, but above
the area that will be immersed in the developing solvent. Allow streaked area
to dry completely.
A.3.3 Streak or spot the standard solution next to the sample spot, at
the same distance from the edge of the plate as the sample spot. Allow streaked
area to dry completely.
A.3.4 Stand the plate in the equilibrated developing tank and cover
tightly. Let the plate develop until capillary action has carried the solvent
nearly to the top of the plate.
NOTE: It may be necessary to adjust the developing time to achieve suitable
separation of the nitroglycerine from interfering sample components.
A.3.5 Remove the plate from the tank and air dry.
A.3.6 Spray the developed plates with a 5% solution of diphenylamine in
ethanol. After spraying developed TLC plates, expose the plates to U.V. light
(longwave, shortwave, or both simultaneously) for 15-30 minutes, until spots
appear. For very weak spots, spray again with 5% diphenylamine in concentrated
sulfuric acid.
A.3.7 Spray developed plates with Greiss reagent (Section A.2.10) and
warm the plates in a 100°C oven for 5-10 minutes.
A.3.8 The presence of a spot at the same distance from the origin as the
standard (Rf of approximately 0.4) indicates that nitroglycerine may be present.
The absence of a spot in this location indicates that nitroglycerine is not
present above the detection limit of the test.
8332-8 Revision 0
Appendix A January 1995
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4.3 DETERMINATION OF ORGANIC ANALYTES
4.3.4 INFRARED METHODS
The following methods are included in this section:
Method 8410: Gas Chromatography/Fourier Transform Infrared
(GC/FT-IR) Spectrometry for Semi volatile
Organics: Capillary Column
Method 8430: Analysis of Bis(2-chloroethyl)ether Hydrolysis
Products by Direct Aqueous Injection GC/FT-IR
Method 8440: Total Recoverable Petroleum Hydrocarbons by
Infrared Spectrophotometry
FOUR - 13 Revision 3
January 1995
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METHOD 8430
ANALYSIS OF BIS(2-CHLOROETHYL) ETHER AND HYDROLYSIS PRODUCTS
BY DIRECT AQUEOUS INJECTION GC/FT-IR
1.0 SCOPE AND APPLICATION
1.1 This method provides procedures for the identification and
quantitation of bis(2-chloroethyl) ether and its hydrolysis compounds in
aqueous matrices by direct aqueous injection (DAI) and gas chromatography with
detection by a fourier transform infrared spectrometer (GC/FT-IR). The
following compounds can be determined by this method:
Compound Name Abbreviation CAS Number3
Bis(2-chloroethyl) ether
2-Chloroethanol
2-(2-Chloroethoxy)ethanol
Diethylene glycol
Ethyl ene glycol
BCEE
CE
2CEE
DEG
EG
111-44-4
107-07-3
628-89-7
111-46-6
107-21-1
a Chemical Abstract Services Registry Number.
1.2 Although the initial study upon which this method is based targeted
only the bis(2-chloroethyl) ether and its hydrolysis compounds, its has been
suggested that this method can be used for the identification of compounds
that are generally non-extractable, highly water soluble, thermally stable,
and do not co-elute with water from the GC. Possible analytes include ethers
and alcohols.
1.3 The minimum identifiable quantities (MIQ) for the five compounds in
organic-free reagent water range from a low of 46 ng for BCEE, to a high of
120 ng for EG (See Sec. 9.2 for MIQ definition). The MIQ for a specific
sample may differ depending on the nature of the interferences in the sample
matrix and the amount of sample used for the analysis. The method detection
limit (MDL) has not yet been determined for the target analytes.
1.4 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatography (GC), the interpretation
of FT-IR spectra and the use of continuous data collection systems. Each
analyst must demonstrate the ability to generate acceptable results with this
method.
2.0 SUMMARY OF METHOD
2.1 Water samples are filtered through a 0.45 jum filter, and 1 /iL
aliquots are injected directly into a GC. The GC is equipped with 2
8430 - 1 Revision 0
January 1995
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detectors, a thermal conductivity detector (TCD) and an FT-IR. During
analysis, the analyst disconnects the FT-IR from the system to prevent aqueous
degradation of the KBr window, using an 8 port switching valve. Further
analysis of either the solid precipitate from the filtering step or the
aqueous filtrate for trace amounts of non-water soluble compounds may be done
by extracting the samples using appropriate 3500 series methods.
3.0 INTERFERENCES
3.1 Method interference may be caused by contaminants in solvents,
reagents, glassware and other sample processing hardware that lead to discrete
artifacts and/or elevated baselines in the chromatograms. All of these
materials must be routinely demonstrated to be free from interferences under
the conditions of the analysis by running laboratory method blanks.
3.1.1 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. It should then be
drained dry, and heated in a laboratory oven at 130°C for several hours
before use. Solvent rinsing with methanol may be substituted for the
oven heating. After drying and cooling, glassware should be stored in a
clean environment to prevent any accumulation of dust or other
contaminants.
3.1.2 The use of high purity reagents and solvents helps to
minimize interference problems. Purification of solvents by
distillation in all-glass systems may be required.
3.2 Matrix interferences may be caused by contaminants that are in the
sample. The extent of matrix interferences will vary considerably from source
to source, depending upon the nature and diversity of the matrix being
sampled. If significant interferences occur in subsequent samples, additional
cleanup may be necessary.
3.3 The extent of interferences that may be encountered in this method
has not been fully assessed. Although the GC conditions described allow for a
unique resolution of the specific compounds covered by this method, other
matrix components may interfere.
4.0 APPARATUS AND MATERIALS
4.1 GC/FT-IR System
4.1.1 GC - Temperature programmable oven equipped with a cool
on-column injection port, a purged splitless injection port, or an
equivalent suitable for capillary glass columns.
4.1.2 Column - 30 m DB-wax, 1.0 p,m film, Megabore (J&W
Scientific), or equivalent.
8430 - 2 Revision 0
January 1995
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4.1.3 Detectors
4.1.3.1 Thermal Conductivity Detector (TCD) - Must be
able to handle temperatures up to 300°C.
4.1.3.2 FT-IR Spectrometer - System should be equipped
with a mercury-cadmium-telluride detector, using a light-pipe
interface with KBr windows (available from Digilab Model GC/C32
or equivalent). Resolution of 8 cm"1 and a range of 4,000 to
7,000 cm"1 is required. The light-pipe interface and transfer
lines should be contained in a heated system (up to 250°C) to
prevent sample condensation. Extra transfer lines will be
needed for the switching system.
4.1.4 Detector Switching System - 8 port stainless steel GC
rotary switching valve, with an inert interval surface (such as FEP
Teflon) and capable of withstanding temperatures up to 300°C. Ideally,
the valve should be mounted inside the GC oven with external control.
If internal mounting is not possible then use of an external heated
valve enclosure may be employed. This allows direct injection of
aqueous samples by using the valve system to route water away from the
FT-IR KBr window which water rapidly destroys.
4.1.5 Data Collection - Each detector should have its own
signal recorder.
4.1.5.1 TCD Signal - Either an analog strip chart
recorder or a digital computerized data collection system is
acceptable.
4.1.5.2 FT-IR Signal - The continuous collection of the
spectrometer's signal will require a computerized data system
with the ability to continuously collect spectra at a rate of 4
scans/sec and add the 4 scans to produce a data point for each
second. The ability to compare the required spectra to a
library of spectra may also be useful for confirmation purposes.
4.2 Glassware
4.2.1 Glass Funnels
4.2.2 Volumetric Flasks (square) - various sizes.
4.2.3 Pipettes (A grade) - various sizes.
4.2.4 Sample Vial with teflon lined lids.
4.3 Syringes - 10 ^l, suitable for GC work.
4.4 Analytical Balance, accurate to ±0.0001 g.
4.5 Vacuum Filtration Apparatus - 0.45 jttm filter and clean glass
flasks that are able to hold at least 1 L of liquid.
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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 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 Helium Gas - Suitable for gas chromatography.
5.4 Methanol, CH3OH - Pesticide reagent grade, nanograde, or
equivalent.
5.5 Stock Standard Solutions
5.5.1 Prepare, in organic-free reagent water, a stock standard
solution containing all of the target analytes at a concentration of
1000 mg/L. Record the actual weight of each compound added and
calculate the actual concentration of each component of the solution.
5.5.2 Transfer the stock standard solution into a Teflon-sealed
screw-cap bottle for storage. Store at 4°C and protect from light.
Check the solution periodically for signs of degradation or evaporation.
This solution must be replaced every 3 months, or when any sign of
degradation or evaporation is observed.
5.6 Calibration Standards
5.6.1 Calibration standards at 500, 250, 100, 50, and 25 mg/L,
from the aqueous stock standard solution, by appropriate volumetric
dilutions with water are suggested. Store as in Sec. 5.5.2. The
calibration solutions should not be made by serial dilution of a single
solution.
5.6.2 Since the MDL for the target compounds in water has not
been established, the suggested calibration curve concentrations may be
modified, depending on matrix interferences and sensitivity of
equipment. Generally speaking, the calibration curve should span at
least one order of magnitude and the working range should bracket the
analyte concentration.
5.7 Spiking Solution - The analyst should monitor the performance of
the analytical system and the effectiveness of the method in dealing with each
sample matrix, by spiking a known amount of the target analytes into blanks or
into matrix spike samples. A suggested preliminary concentration of the
spiking stock solution is 100 g/L. When this method becomes better
established, the spiking concentration should be set at 1 to 5 times higher
than the background concentration determined for that matrix.
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5.8 Surrogate Spiking Solution: To monitor the performance of the
method for all samples, a minimum of one surrogate compound should chosen by
the laboratory. This compound should be diluted with water to an appropriate
concentration and added to all samples, method blanks, matrix spikes, and
calibration standards.
6.0 SAMPLE COLLECTION, PRESERVATION AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
6.2 Samples should be stored at 4°C and protected from light.
6.3 Samples should be filtered and analyzed within 14 days of
collection.
7.0 PROCEDURE
7.1 GC/FT-IR Conditions (Recommended):
Flow Rate: 2.4 mL/minute of helium carrier gas.
Run Time: approximately 15 minutes
Injection Volume: 1 jitL
Valve Switch Time: 4 minutes
Temperature Program: 80"C to 220°C at 15°C/minute, hold at 220°C
for 10 minutes.
TCD Temperature: 290°C
GC/FT-IR Interface,
Transfer Lines, and
Light Pipe Temperature: 220°C
Scan Time: 4 scans/second
NOTE: Modifications of these parameters may be necessary to facilitate the
separation of certain compounds depending on matrix interferences
encountered.
7.2 External Calibration
7.2.1 Calibration standards may be prepared using the suggested
concentrations in Sec. 5.6. Matrix interferences may prevent
quantitation at the suggested concentrations. When necessary, the
lowest concentration of the calibration curve should be adjusted to be
at, or near, the method detection limit. Refer to Method 8000, Sec. 7.0
for proper external calibration procedures.
7.2.2 Refer to Method 8000, Sec. 7.0 for the establishment of
retention time windows.
7.2.3 Analyze a solvent blank to ensure that the system is
clean and interference free.
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7.2.4 Analyze the 5 calibration standards, starting with the
lowest concentration and ending with highest concentration.
7.2.5 Tabulate the IR absorbance peak area along with the
calibration factor (CF) for the analyte at each concentration. Refer to
Sec. 7.0 of Method 8000 for linear and non-linear calibration acceptance
criteria. It should be noted that IR transmission is not directly
proportional to concentration.
7.2.5 Recheck the instrument calibration each day, before and
after an analysis is performed, by analyzing one or more calibration
standards. The response obtained should fall within ±15 percent of the
expected value or the instrument must be recalibrated.
7.2.6 After the analysis of 10 or fewer samples, one of the
calibration standards must be reanalyzed to ensure that the retention
times and the CFs of the target analytes remain within the QC
requirements.
7.3 Sample Spiking and Filtering
7.3.1 Allow the sample to come to ambient temperature. Mark
the water meniscus on the side of the 1 L sample bottle for later
determination of exact sample volume.
7.3.2 Add 2 ml of the spiking solution to the spiked blank and
the matrix spike sample. The final concentration of the added analytes
should be about 200 mg/L.
7.3.3 Vacuum filter the sample through a 0.45 /urn filter that
has been rinsed with organic-free reagent water. The filtrate should be
collected in a clean glass bottle. Any particulate collected by the
filtering process may be discarded or extracted for analysis of trace
amounts of non-water soluble compounds using an appropriate 3500 series
method.
7.3.4 Using a pipette, withdraw a 5 ml aliquot of the sample
(aqueous filtrate) and place it into a clean glass sample vial with a
teflon lined lid. This 5 ml aliquot will be the sample used for the
direct aqueous injection of the water sample. Store at 4°C in the dark.
The remainder of the aqueous filtrate may be saved in a glass bottle
with a teflon lined lid until after the analysis is complete or
extracted for analysis of trace amounts of non-water soluble compounds
using an appropriate 3500 series method.
7.4 GC 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.
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7.4.2 Determining the Valve Switching Time - The valve switch
time must be determined before proceeding with the analysis of samples.
7.4.2.1 Place the GC switching valve in the position
which routes the sample to the TCD and away from the FT-IR
spectrometer and inject a 1 ^L aliquot of a reagent blank to
determine the retention time of water in the system. Use this
retention time to determine the optimum valve switch time to
cycle water away from the KBr window loop. At the optimum valve
switch time, the 8 port valve may be moved to the position which
allows use of the FT-IR detector and the TCD in tandem after the
bulk of the aqueous portion of the sample has been diverted away
from the KBr window (See Figure 1).
7.4.2.2 Once the optimum valve switch time is
established, the system may be re-evaluated with one of the
aqueous calibration standards to assure complete separation of
all target analytes.
NOTE: Traces of water in the GC/FT-IR interface are acceptable. However, the
repeated injection of 1.0 jiL aqueous samples may eventually destroy the
KBr window used for FT-IR detection. Care should be taken in deciding
the valve switching time so that the KBr window is exposed to only trace
amounts of water and target analytes that elute with retention times
just after water are not missed.
7.4.2.3 If the TCD is proven to have adequate
sensitivity for a particular analysis, it may be used as the
prime detector once the target analytes are identified using the
FT-IR detector. When the TCD is used as the prime detector, the
switching valve should remain in the position which diverts the
sample away from the KBr window.
7.4.3 Sample Analysis - Analyze the samples, blanks, spiked
blanks, and spiked matrix samples by injecting 1 juL aliquots into the
GC and switching to the FT-IR spectrometer at the previously determined
time. Dilution of the sample may be necessary to adjust the analyte
concentration to within the working range of the calibration curve. If
dilution is necessary, note which samples were diluted in the final
report and make the appropriate calculation adjustments.
7.4.4 GC/FT-IR Identification - Visually compare the analyte
infrared (IR) spectrum versus the search library spectrum of the most
promising on-line library search hits. Report, as identified, those
analytes with IR frequencies for the five (maximum number) most intense
IR bands (S/N > 5) which are within ±5.0 cm"1 of the corresponding bands
in the library spectrum. Choose IR bands which are sharp and well
resolved. The software used to locate spectral peaks should employ the
peak "center of gravity" technique. In addition, the IR frequencies of
the analyte and library spectra should be determined with the same
computer software.
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7.5 Calculations
7.5.1 Calculate the calibration factor for each calibration
standard and determine the percent relative standard deviation (%RSD)
using the external standard calibration procedure in Sec. 7.0 of Mehtod
8000.
7.5.2 Calculation of the concentration of analytes using the
external standard calibration procedure is provided in Sec. 7.0 of
Method 8000.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures. Quality control procedures to validate sample extraction is
covered 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 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. Refer to Appendix A for
FT-IR spectrometer QC requirements.
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 a method blank, matrix spike, a duplicate, 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, the 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
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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.
8.4.3 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. Currently, surrogate compounds have not been
selected for this procedure. 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 No MDL data are available.
9.2 Minimum Identifiable Quantities (MIQ)
9.2.1 The MIQ is defined as the minimum quantity that must be
injected to result in a spectral match that has the correct compound
identification in the top 5 spectral matches. The MIQ will vary
depending on instrument sensitivity and sample matrix effects.
9.2.2 The MIQ range for CE, BCEE, EG, 2CEE, and DEG in organic-
free reagent water by direct aqueous injection goes from a low of 46 ng
for BCEE to a high of 120 ng for EG.1
10.0 REFERENCES
1. Payne, W.D. and Collette, T.W., "Identification of Bis(2-chloroethyl)
ether Hydrolysis Products by Direct Aqueous Injection GC/FT-IR," J. of
High Res. Chrom.. 12, 693-696, 1989.
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METHOD 8430
ANALYSIS OF BIS(2-CHLOROETHYL) ETHER AND HYDROLYSIS PRODUCTS
BY DIRECT AQUEOUS INJECTION
7.1 Set operating con-
ditions of the GC/FT-IR
to facilitate seperation
of compounds
7.2 Perform calibration
(calibration should be
checked each day and a
calibration standard should
be frequently analyzed to
ensure retention times and
RFs remain within QC
requirements.)
7.3.1 Allow sample to come
to ambient temperature and
mark the water meniscus on
the sample bottle for later
volume determination.
7.3.2 Add 2 ml of the
spiking solution to the
spiked bland and the
matrix spike sample.
7 .3.3 Filter sample
through a 0.45 /im
filter.
7.3.4 Withdraw a bmL
aliquot and store at 4°C
in the dark (to be used
for DAI of the water sample.)
7.4.1 Refer to Method 8000
for instructions on analysis
sequence, appropriate
dilutions, and mdentification
criteria.
7.4.2 Establish optimum
valve switch time and re-
evaluate the system to
ensure complete separation
of the target analysis.
7.4.3 Establish optional
retention time window
(refer to Method 8000.)
7.4.4 Inject samples,
blanks, spiked blanks,
and spiked matrix samples
and analyze.
7.4.4
Is analyte
concentration
within the working
range of the
calibration'
7.4.4 Dilute and
reanalyze.
745 Visually compare the
analyte IR spectrum to
the search library spectrum.
7.4.5 Report as identified,
those analytes with IR
frequencies within
+ /- 5.0 cml for the five
(maximum number) most
intense corresponding bands
in the library spectrum.
7.5.1 Calculate each
response factor using
equations provided.
7.5.2 Calculate the
concentrations of the
analytes (see Method
8000.)
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APPENDIX A
Quality Control of FT-IR Spectrometer
A.I Equipment Adjustments and Maintenance
A.1.1 Mirror Alignment - Adjust the interferometer mirrors ti attain
the most intense signal. Data collection should not be initiated until the
interferogram is stable. If necessary, align the mirrors prior to each GC/FT-
IR run.
A.1.2 Interferometer - If the interferometer is air-driven, adjust the
interferometer drive air pressure to manufacturer's specifications.
A.1.3 Lightpipe - The lightpipe and lightpipe windows should be
protected from moisture and other corrosive substances at all times. For this
purpose, maintain the lightpipe temperature above the maximum GC program
temperature but below its thermal degradation limit. When not in use,
maintain the lightpipe temperature slightly above ambient. At all times
maintain a flow of dry, inert, carrier gas through the lightpipe.
A.1.4 Beamsplitter - If the spectrometer is thermostated, maintain the
beamsplitter at a temperature slightly above ambient at all times. If the
spectrometer is not thermostated, minimize exposure of the beamsplitter to
atmospheric water vapor.
A.2 Centerburst Intensity and MCT Detector Check
A.2.1 With an oscilloscope, check the MCT detector centerburst
intensity versus the manufacturer's specifications. Increase the source
voltage, if necessary, to meet these specifications. For reference purposes,
laboratories should prepare a plot of time versus detector voltage over at
least a 5 day period.
A.2.2 If the centerburst intensity is 75 percent or less of the mean
intensity of the plot maximum obtained by the procedure, install a new source
and check the MCT centerburst with an oscilloscope versus the manufacturer's
specifications (if available). Allow at least five hours of new source
operation before data acquisition.
A.2.3 Align Test - With the lightpipe and MCT detector at thermal
equilibrium, check the intensity of the centerburst versus the signal
temperature calibration curve. Signal intensity deviation from the predicted
intensity may mean thermal equilibrium has not yet been achieved, loss of
detector coolant, decrease in source output, or a loss in signal throughput
resulting from lightpipe deterioration.
A.3 GC/FT-IR Sensitivity
A.3.1 Capillary Column Interface Sensitivity Test - Install a 30 m x
0.32 mm fused silica capillary column coated with 1.0 /Ltm of DB-5 (or
equivalent). Set the lightpipe and transfer lines at 280°C, the injector at
225°C and the GC detector at 280°C (if used). Under splitless Grob-type or
on-column injection conditions, inject 25 ng of nitrobenzene, dissolved in 1
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fj.L of methylene chloride. The nitrobenzene should be identified by the on-
line library software search within the first five hits (nitrobenzene should
be contained within the search library).
A.3.2 One Hundred Percent Line Test - Set the GC/FT-IR operating
conditions to those employed for the Sensitivity Test (see Sec. A.3.1).
Collect 16 scans over the entire detector spectral range. Plot the test and
measure the peak-to-peak noise between 1800 and 2000 cm"1. This noise should
be less than or equal to 0.15 percent. Store this plot for future reference.
A.3.3 If the GC/FT-IR was purchased before 1985, there may be a
temperature effect at the interface. To account for this, prepare a plot of
lightpipe temperature versus MCT centerburst intensity (in volts or other
vertical height units). This plot should span the temperature range between
ambient and the lightpipe thermal limit in increments of about 20°C. Use this
plot for daily QA/QC (see Sec. A.2.3). Note that modern GC/FT-IR interfaces
(1985 and later) may have eliminated most of this temperature effect.
A.4 Frequency Calibration - At the present time, no consensus exists within
the spectroscopic community on a suitable frequency reference standard for
vapor-phase FT-IR. One reviewer has suggested the use of indene as an on-the-
fly standard.
A.5 Single Beam Test - With the GC/FT-IR at analysis conditions, collect 16
scans in the single beam mode. Plot the co-added file and compare with a
subsequent file acquired in the same fashion several minutes later. Note if
the spectrometer is at purge equilibrium. Also check the plot for signs of
deterioration of the lightpipe potassium bromide windows. Store this plot for
future reference.
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METHOD 8440
TOTAL RECOVERABLE PETROLEUM HYDROCARBONS BY INFRARED SPECTROPHOTOMETRY
1.0 SCOPE AND APPLICATION
1.1 Method 8440 (formerly Method 9073) is used for the measurement of
total recoverable petroleum hydrocarbons (TRPHs) extracted with supercritical
carbon dioxide from sediment, soil and sludge samples using Method 3560.
1.2 Method 8440 is not applicable to the measurement of gasoline and
other volatile petroleum fractions.
1.3 Method 8440 can detect TRPHs at concentrations of 10 mg/L in
extracts. This translates to 10 mg/Kg in soils when a 3 g sample is extracted
by SFE (assuming 100 percent extraction efficiency), and the final extract
volume is 3 mL.
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 Soil samples are extracted with supercritical carbon dioxide using
Method 3560. Interferences are removed with silica gel, either by shaking the
extract with loose silica gel, or by passing it through a silica gel solid-
phase extraction cartridge. After infrared (IR) analysis of the extract,
TRPHs are quantitated by direct comparison with standards.
3.0 INTERFERENCES
3.1 The analyte class being measured (TRPHs) is defined within the
context of this method. The measurement may be subject to interferences, and
the results should be interpreted accordingly.
3.2 Determination of TRPHs is a measure of mineral oils only, and does
not include the biodegradable animal greases and vegetable oils captured in
oil and grease measurements. These non-mineral-oil contaminants may cause
positive interferences with IR analysis, if they are not completely removed
by the silica gel cleanup.
3.3 Method 8440 is not appropriate for use in the analysis of gasoline
and other volatile petroleum fractions because these fractions evaporate
during sample preparation.
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4.0 APPARATUS AND MATERIALS
4.1 Infrared spectrophotometer - Scanning or fixed wavelength, for
measurement around 2950 cm"1.
4.2 IR cells - 10 mm, 50 mm, and 100 mm pathlength, sodium chloride or
IR-grade glass.
4.3 Magnetic stirrer with Teflon-coated stirring bars.
4.4 Optional - A vacuum manifold consisting of glass vacuum basin,
collection rack and funnel, collection vials, replaceable stainless steel
delivery tips, built-in vacuum bleed valve and gauge is recommended for use
when silica gel cartridges are used. 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.
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 Hydrochloric acid (1:1 v/v), HC1. Mix equal volumes of
concentrated HC1 and water.
5.4 Tetrachloroethylene, C2C14 - spectrophotometric grade, or
equivalent.
5.5 Raw materials for reference oil mixture - spectrophotometric grade,
or equivalent.
5.5.1 n-Hexadecane, CH3(CH2)14CH3
5.5.2 Isooctane, (CH3)3CCH2CH(CH3)2
5.5.3 Chlorobenzene, C8H5C1
5.6 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 (CH2C12). If the sodium sulfate is precleaned with
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methylene chloride, a method blank must be analyzed, demonstrating that there
is no interference from the sodium sulfate.
5.7 Silica gel.
5.7.1 Silica gel solid-phase extraction cartridges (40 /urn
particles, 60 A pores), 0.5 g, Supelco, J.T. Baker, or equivalent.
5.7.2 Silica gel, 60 to 200 mesh, Davidson Grade 950 or
equivalent (deactivated with 1 to 2 percent water).
5.8 Calibration mixtures:
5.8.1 The material of interest, if available, or the same type
of petroleum fraction, if it is known and original sample is
unavailable, shall be used for preparation of calibration standards.
Reference oil is to be used only for unknowns. Whenever possible, a GC
fingerprint should be run on unknowns to determine the petroleum
fraction type.
5.8.2 Reference oil - Pipet 15.0 ml n-hexadecane, 15.0 ml
isooctane, and 10.0 ml chlorobenzene into a 50 ml glass-stoppered
bottle. Maintain the integrity of the mixture by keeping stoppered
except when withdrawing aliquots. Refrigerate at 4°C when not in use.
5.8.3 Stock standard - Pipet 0.5 ml calibration standard (Section
5.8.1 or 5.8.2) into a tared 100 mL volumetric flask and stopper
immediately. Weigh and dilute to volume with tetrachloroethylene.
5.8.4 Working standards - Pipet appropriate volumes of stock
standard (Sec. 5.8.3) into 100 ml volumetric flasks according to the
cell size to be used. Dilute to volume with tetrachloroethylene.
Calculate the concentrations of the standards from the stock standard
concentrations.
5.8 Calibration mixture for silica gel cleanup - Prepare a stock
solution of corn oil by placing about 1 ml (0.5 to 1 g) of corn oil into a
tared 100 ml volumetric flask. Stopper the flask and weigh to the nearest
milligram. Dilute to the mark with tetrachloroethylene, and shake the
contents to effect dissolution. Prepare additional dilutions to cover the
range of interest.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Solid samples should be collected and stored as any other solid
sample containing semivolatile analytes. See the introductory material to
this Chapter, Organic Analytes, Sec. 4.1.
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6.2 Samples should be analyzed with minimum delay, upon receipt in the
laboratory, and must be kept refrigerated prior to analysis.
7.0 PROCEDURE
7.1 Prepare samples according to Method 3560.
7.2 Add 0.3 g of loose silica gel to the extract and shake the mixture
for 5 minutes, or pass the extract through a 0.5 g silica gel solid-phase
extraction cartridge (conditioned with 5 ml of tetrachloroethylene). When
using loose silica gel, filter the extract through a plug of precleaned
silanized glass wool in a disposable glass pipette.
7.3 After the silica gel cleanup, fill a clean IR cell with the
solution and determine the absorbance of the extract. If the absorbance
exceeds the linear range of the IR spectrophotometer, prepare an appropriate
dilution and reanalyze. The possibility that the absorptive capacity of the
silica gel has been exceeded can be tested at this point by repeating the
cleanup and determinative steps.
7.4 Select appropriate working standard concentrations and cell
pathlengths according to the following ranges:
Concentration range
Pathlength (mm) fug/ml of extract) Volume (ml)
10 5 to 500 3
50 1 to 100 15
100 0.5 to 50 30
Calibrate the instrument for the appropriate cells using a series of
working standards. Determine absorbance directly for each solution at the
absorbance maximum at about 2950 cm"1. Prepare a calibration plot of
absorbance versus concentration of petroleum hydrocarbons in the working
standards.
7.5 Determine the concentration of TRPHs in the extract by comparing the
response against the calibration plot.
7.6 Calculate the concentration of TRPHs in the sample using the
formula:
R x D x V
Concentration (mg/Kg) =
W
where:
R = mg/mL of TRPHs as determined from the calibration plot
V = volume of extract, in milliliters
D = extract dilution factor, if used
W = weight of solid sample, in kilograms.
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7.7 Recover the tetrachloroethylene used in this method by distillation
or other appropriate technique.
8.0 QUALITY CONTROL
8.1 Reagent blanks or matrix-spiked samples must be subjected to the
same analytical procedures as those used with actual samples.
8.2 Refer to Chapter One for specific Quality Control procedures and to
Method 3500 for sample preparation procedures.
9.0 METHOD PERFORMANCE
9.1 Table 1 presents a comparison of certified values and the values
obtained using Methods 3560 and 8440. Data are presented for both Freon-113
and tetrachloroethylene, since both solvents were found to be an acceptable
collection solvent. However, only tetrachloroethylene is recommended as a
collection solvent for TRPHs in Method 3560.
9.2 Table 2 presents precision and accuracy data from the single-
laboratory evaluation of Methods 3560 and 8440 for the determination of
petroleum hydrocarbons from spiked soil samples. These data were obtained by
extracting samples at 340 atm/80°C/60 minutes (dynamic).
10.0 REFERENCES
1. Rohrbough, W. G.; et al. Reagent Chemicals, American Chemical Society
Specifications, 7th ed.; American Chemical Society, Washington, DC, 1986.
2. Methods for Chemical Analysis of Water and Wastes; 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, 1983; EPA-600/4-79-
020.
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TABLE 1
CERTIFIED AND SPIKE VALUES COMPARED TO RESULTS
OBTAINED BY METHODS 3560/8440
Reference Material
Environmental Resource Assoc.
TPH-1 (lot 91012)
Environmental Resource Assoc.
TPH-2 (lot 91012)
Clay spiked with keroseneb
Clay spiked with light gas oil0
Clay spiked with heavy gas oild
Environmental Resource Assoc.
TPH-1 (lot 91017)"
Environmental Resource Assoc.
TPH-2 (lot 91017)6
Spike cone, or
certified cone.
(mg/kg)
1,830
2,230
100
100
100
614
2,050
Methods
3560/8440
(mg/kg)
l,920±126a
2,150±380a
86.0; 93.0
84.0; 98.0
103; 108
562; 447
1,780; 1,780
8 Three 60 minute extractions. The extracted material was collected in
Freon-113; the concentrations were determined against the reference oil
standard.
b Duplicate 30 minute extractions. The extracted material was collected in
tetrachloroethylene; the concentrations were determined against standard
made from the spiking material.
0 Six 30 minute extractions. The extracted material was collected in
tetrachloroethylene; the concentrations were determined against a standard
made from the spiking material.
d Four 30 minute extractions. The extracted material was collected in
tetrachloroethylene; the concentrations were determined against a standard
made from the spiking material.
6 Three 30 minute extractions. The extracted material was collected in
tetrachloroethylene; the concentrations were determined against the
reference oil standard.
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TABLE 2
SINGLE-LABORATORY METHOD ACCURACY AND PRECISION FOR
METHODS 3560/8440 FOR SELECTED MATRICES
Spike cone, or Method
certified cone. Spike accuracy
Matrix (mg/kg) Material (% recovery)
Clay soil6
ERA TPH-18
(lot 91016)
ERA TPH-28
(lot 91016)
SRS103-100b
2,500 Motor oil 104
2,350 Vacuum oil 80.3
1,450 Vacuum oil 88.6
32,600 c 94.2
Method
precision
(% RSD)
8.5
19.7
19.6
4.0
Eight determinations were made using two different supercritical fluid
extraction systems. The extracted material was collected in Freon-113.
Ten determinations were made using three different supercritical fluid
extraction systems. The extracted material was collected in
Freon-113.
This is a standard reference soil certified for polynuclear aromatic
hydrocarbons. No spike was added.
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METHOD 8440
TOTAL RECOVERABLE PETROLEUM HYDROCARBONS BY INFRARED SPECTROPHOTOMETRY
Prepare sample(s)
according to
appropriate Method.
7.5 - 7.6 Calculate
the concentration of
TPHs in the sample.
7.7 Recover the
tetrachloroethylene
by appropriate
technique.
7.2 Proceed with
silica gel cleanup
7.3 Determine the
absorbance of
the extract.
7.3
Does
absorbance excee
linear range
of the IR
spectro-
photometer
7.3 Dilute
appropriately
7.4 Calibrate the
instrument.
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4.3 DETERMINATION OF ORGANIC ANALYTES
4.3.5 MISCELLANEOUS SPECTROMETRIC METHODS
The following method is included in this section:
Method 8520: Continuous Measurement of Formaldehyde in Ambient
Air
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METHOD 8520
CONTINUOUS MEASUREMENT OF FORMALDEHYDE IN AMBIENT AIR
1.0 SCOPE AND APPLICATION
1.1 This method is applicable to the continuous measurement of
formaldehyde (CAS No. 50-00-0) in ambient air. This method is for use primarily
for nonoccupational exposure monitoring.
1.2 This method is applicable to concentrations of formaldehyde from 6 to
500 p-g/m3. Detection limits are dependent on sample airflow rate, with a maximum
rate set at 1.0 L/min.
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 A modified portable commercial analyzer which consists of a small
diaphragm pump, an analytical module, a colorimeter with glass flow cells, a
peristaltic pump and tubing, and a 12 volt rechargeable battery for optional DC
power operation is used for this method (See Figure 1). An acidified
pararosanil ine scrubber solution is pumped through the reference cell in the
colorimeter and is then diluted 1:1 with a liquid flow stream containing water.
The diluted solution then passes into an air scrubber coil where formaldehyde is
quantitatively absorbed from the air sample stream. After the liquid and air are
separated, the sample solution is mixed 1:1 with a dilute aqueous Na2S03 reagent.
A series of time delay coils allow 16 minutes for color development of the
chromophore. The colored product is continuously detected as it passes through
the sample cell by a UV-Visible spectrophotometer set at 550 nm and is recorded
on a strip chart recorder or an automated data acquisition system.
3.0 INTERFERENCES
3.1 Method interferences may be caused by contaminants in reagents,
glassware, and sample processing hardware.
3.1.1 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. It should then be drained dry,
and heated in a laboratory oven at 130°C for several hours before use.
Solvent rinsing may be substituted for the oven heating. After drying and
cooling, glassware should be stored in a clean environment, covered with
aluminum foil to prevent any accumulation of dust or other contaminants.
3.1.2 All of these materials and equipment must be routinely
demonstrated to be free from interferences under the conditions of the
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analysis by establishing the analyzer baseline with the introduction of a
source of zero air (zero grade compressed air) at the sample inlet.
3.1.3 Instrument Cleaning
3.1.3.1 Reagent Lines - To ensure optimum performance of the
analyzer, the three reagent lines should be placed in a reservoir
containing 1.0 M nitric acid as a cleaning solution. Operate the
analyzer with the zero air supply and pump the cleaning solution
through the analyzer for 1 hour. To rinse the lines, pump organic-
free reagent water through the system for 2 hours. The cleaning
sequence must be performed at frequent intervals (e.g. each second
or third day of regular use).
3.1.3.2 Colorimeter Flow Cell - Clean the flow cells by
periodically soaking them in commercial bleach or a laboratory
detergent solution. After cleaning the cells, rinse them well with
organic-free reagent water.
3.2 Although there is some disagreement concerning the mechanism
involving the formation of the final product in the reaction between
pararosaniline and formaldehyde, it is generally agreed that the chromophore is
an alkylsulfonic acid. (Refs. 1,3) Studies have shown that the color-forming
reaction of the modified pararosaniline procedure is highly specific to
formaldehyde.
3.2.1 Low molecular weight aldehydes exhibit positive
interferences, but only when present in large excess over formaldehyde.
3.2.2 Sulfur dioxide, at a concentration of 520 M9/m3, produces a
slight negative interference.
3.3 Sensitivity and color development time are functions of temperature.
Good results may be obtained for operating temperatures between 15 and 35°C (with
optimum results at 25°C) by performing daily instrument calibrations.
3.4 Baseline drift is negligible, or less than ±2% of full scale after
8 hours of continuous operation, under normal conditions. Periodic baseline
checks should be performed to ensure accuracy.
3.5 Low-concentration aqueous formaldehyde standard solutions of 0.02 to
2.0 /jg/mL and dilute sodium sulfite reagent solutions exhibit a limited
stability and therefore must be prepared daily.
3.6 The use of high purity reagents and solvents helps to minimize
interference problems.
4.0 APPARATUS AND MATERIALS
4.1 Continuous Air Monitor
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4.1.1 Automated Wet-chemical Colorimetric Analyzer - CEA
Instruments Model TGM 555 air monitor equipped with AMK-4 formaldehyde
analytical module or equivalent. The TGM 555 analyzer is equipped with a
dual beam colorimeter, set at 550 nm, consisting of glass reference and
sample flow cells, a matched pair of photodetectors, a miniature tungsten-
halogen lamp, and a signal processing unit. The colorimeter measures the
difference in light absorption of the pararosaniline chloride reagent
before and after it reacts with formaldehyde.
4.1.2 Spare Parts - A spare parts maintenance kit is available from
CEA or the following equipment may be purchased.
4.1.2.1 Silicon rubber peristaltic pump tubes (T-78) - Pump
tubes should be replaced after 30 to 45 days of continuous use.
4.1.2.2 Rubber diaphragm in the miniature air pump - The
diaphragm should be replaced whenever it becomes worn or defective.
4.1.2.3 Tungsten-halogen colorimeter lamp
4.1.3 Glass Coils (3 mm OD) and Connectors - These are required for
modifications of the analytical module to increase residence time for the
reactants. The standard version of the AMK-4 analytical module (2 x 25
turn coil) contains a residence mixing time of less than 6 minutes, as
measured from the glass double mixing tee to the entrance of the sample
cell. By adding additional mixing coils (an additional 120 turns) the
residence mixing time was increased to 16 minutes (absorbance reached a
maximum when the residence time of the reactants was about 16 minutes or
longer). These parts are used extensively in segmented flow colorimetric
analyzers such as Technicon Instruments Corporations Autoanalyzer II or an
equivalent.
4.1.4 Glass Debubbler - A second glass debubbler was added just
ahead of the entrance to the sample cell to overcome the increased
frictional drag on the system. The top exit of the debubbler was attached
to a T-78 silicon rubber tube on the peristaltic pump so that all of the
air between the sample segments is drawn away to waste and approximately
95% of the sample liquid is pushed into the sample cell for measurement.
Further, by inverting the glass double mixing tee so that the sample
stream enters from the top and the Na2S03 reagent enters from below,
smoother operation (i.e., a more regular bubble pattern) resulted. All
connections from the exit of the glass double mixing tee to the entrance
of the sample cell are 3 mm OD glass tubing.
4.2 Zero Air Supply
4.2.1 A regulated, zero grade compressed air cylinder with
connections to the instrument sample inlet through a tee fitting to permit
atmospheric dumping of the excess air supply.
4.2.2 Alternatively, formaldehyde-free air may be obtained by
scrubbing ambient air through a series of 3 midget impingers, each
containing 15 ml of 0.05 M sulfuric acid, followed by a silica-gel packed
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cartridge for drying the airstream. The impinger solutions should be
replaced daily.
NOTE: Avoid the use of organic vapor filters which may be supplied with a
commercial instrument. These filters have proven to be unsuitable for
use at formaldehyde concentrations less than 500
4.2.3 Teflon Tubing and Valves - FEP Teflon tubing with a
0.25 in. OD and a 3-way Teflon valve for switching (manual or solenoid)
between the zero air supply and the ambient air sample stream.
4.3 Data Acquisition System - Data acquisition may be accomplished with
a manual strip chart recorder or an automated data acquisition system.
4.4 Glassware
4.4.1 Volumetric Flasks, class A - 1 L, 250 ml, and other various
sizes.
4.4.2 Pipets, class A - various sizes
4.4.3 Mohr Pipets - 1.0 ml with 0.01 ml graduations
4.4.4 Soap Bubble Flowmeter - 500 ml
4.5 Thermometer - to check the temperature at the time of sampling.
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 Formaldehyde, CH20 - ACS certified or assayed 36.5% solution (w/w),
store at room temperature and protect from light.
5.4 Hydrochloric acid solution, HC1 in water - 2.0 M
5.5 Nitric Acid solution, HN03 in water - 1.0 N
5.6 Sodium sulfite, Na2S03 - ACS certified, anhydrous crystal
5.7 2.78 mM Sodium sulfite solution - Dissolve 0.175 anhydrous sodium
sulfite crystal in 500 ml of water. This solution should be protected from light
during use and must be prepared daily.
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5.8 Pararosaniline chloride (Basic Fuchsin), C20H20N3C1 - specially
purified, 0.2% (w/v) in 1 M HC1 (Fisher Scientific, or equivalent).
5.9 1.72 mM Pararosaniline chloride in 0.8 M HC1
5.9.1 Using a 100 ml graduated cylinder, measure and combine
68.5 ml of the stock pararosaniline chloride solution with 66 ml of 2.0 M
HC1 solution in a 250 ml volumetric flask.
5.9.2 Dilute the mixture with water to a final volume of 250 ml.
5.9.3 This solution should be protected from light during use. The
solution may be stored up to 3 months if kept at 4"C and protected from
light.
NOTE: The rate of consumption of the 2.78 mM sodium sulfite solution and the
1.72 mM pararosaniline chloride solution is 0.6 L per 24 hours at routine
analyzer operation rate of 0.40 mL/min. After mixing the three solutions
together with glass double mixing tee, the final concentration of the
reactants should be 0.43 mM Pararosaniline, 0.2. M HC1, and 1.39 mM Na2S03.
5.10 Formaldehyde Stock Standard Solution - Add approximately 2.1 g of 37%
formaldehyde solution, weighed to the nearest ±0.1 mg, to a 1 L volumetric flask
and dilute to the mark to make a solution of approximately 777 ng/ml. Determine
the actual concentration of the solution using Method 8315. The stock standard
formaldehyde solution is stable for up to 1 year if it is protected from the
light and stored at 4°C.
5.11 Secondary Formaldehyde Standard Solutions - Prepare 2 more solutions
by two 25:1 serial dilutions of the stock standard solution with water to
approximately 31.1 and 1.24 p.g/m\.. If the concentration of the formaldehyde
stock solution is not 777 /ug/ml_, the actual concentration of the secondary
standards must be calculated. If the solution are kept in the dark and at 4'C,
the 31.1 jig/mL standard may be stored for 3 months and the 1.24 jug/mL standard
may be stored for 7 days.
5.12 Formaldehyde Calibration Standard Solutions - Prepare 3 calibration
standards with concentrations ranging from approximately 0.03 to 1.24 jug/mL.
Formaldehyde concentration in the above range are equivalent to air
concentrations from 11.2 to 492 jug/m3. Working standards must be prepared daily.
6.0 SAMPLE COLLECTION
6.1 Start-up Procedure
NOTE: The signal response for a 100 ppbv liquid standard remained unchanged for
the temperatures in the 20-30'C range. Check the temperature before
analysis to assure environment to be analyzed is within the proper
temperature range.
6.1.1 Activate the AC power switch and the DC power switch which
supplies power to the colorimeter lamp and LED digital display panel.
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Power to the colorimeter lamp should be left on continuously during
periods of regular use to maintain the thermal stability of the unit.
6.1.2 Allow a minimum 2 hour warm-up period before starting the
analyzer. During periods of intermittent regular use, keep the AC and DC
power switches on continuously, in order to minimize start-up time.
6.1.3 Set the Range switch to the "low" position and the Damp
switch to the "high" position.
6.1.4 Install the 7 peristaltic pump tube lines in their proper
sequence on the pump rollers. Place the 3 reagent lines in a water
reservoir, then switch the sample air inlet valve to the zero air supply
position.
6.1.5 Activate the Liquid Flow switch, to start the reagent pump
and, at the same time, turn on the Air Flow switch to start the air pump
(using the zero air supply). Pump water for 20 to 30 minutes to allow the
system flows to stabilize and to verify trouble-free operation.
6.1.6 Deactivate the liquid flow pump and the airflow pump. Place
each of the reagent lines in its proper reagent reservoir and restart both
pumps. Again, using the zero air supply, allow the reagents to pump
through the analyzer for 20 to 30 minutes.
6.1.7 Follow the operational manual procedure for manual adjustment
of the optical zero on the colorimeter (if necessary). Use the zero
potentiometer to make fine adjustments for setting the baseline response
to a zero setting on the display panel. The span potentiometer should be
adjusted to the maximum (10.0) setting for most low-concentration
formaldehyde measurements.
6.1.8 Activate the strip chart recorder (or data acquisition
system) and adjust the baseline response to read 5 to 10 percent of the
full scale response. The selection of the full-scale voltage output range
depends on the sensitivity required for the particular monitoring
application. A 1.0 volt signal response corresponds to approximately
120 M9/m3 formaldehyde in the sample airstream.
6.1.9 Monitor the baseline response of the instrument for 10 to 15
minutes to verify that the signal is stable.
6.2 Routine Operating Procedure
6.2.1 After the instrument baseline has stabilized using the zero
air supply, initiate the multipoint calibration procedure (as in Sec.
7.1.3) using the fresh liquid equivalent formaldehyde standard solutions
from Sec. 5.11.
6.2.2 After sampling the final liquid standard in the calibration
sequence, return the reagent line that is pumping the calibration
solutions to the water reservoir and reestablish the instrument baseline.
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6.2.3 Record the chart speed and the exact starting time. Begin
monitoring the sample atmosphere by switching the sample air inlet valve
from the zero air supply to the air sample position. After a lag time of
18 minutes, the instrument will respond quickly (1 to 2 minutes) to the
concentration of formaldehyde in the airstream.
6.2.4 Conduct periodic baseline and calibration checks at intervals
appropriate to the particular operating characteristics of each analyzer
system (see Sec. 7.1.3) and the specific monitoring application.
6.3 Shutdown Procedure
6.3.1 With the sample inlet valve in the zero air supply position
and with the monitor in operation, place all 3 of the reagent lines in a
reservoir containing water.
6.3.2 Pump water through the instrument for 30 minutes, then, shut
off both the liquid flow pump and the airflow pump. Remove the 7 pump
tubes from the peristaltic pump rollers if the instrument is to remain
shut down for more than 3 hours.
6.3.3 Deactivate the zero air supply (if necessary) and secure the
reagent reservoirs for proper storage. Properly dispose of the contents
of the waste reservoir in accordance with acceptable environmental
procedures.
6.3.4 Keep the colorimeter lamp on unless the instrument is to
remain inactive for 7 days or more.
6.4 Analysis of Data
6.4.1 Determine the height of each peak or plateau in the analysis
data by measuring from the established baseline.
6.4.2 Determine the time at which any peaks or plateaus occurred
during the sampling analysis period.
6.4.3 Using the equation of the calibration curve determined in
Sec. 6.2.1, calculate the concentration of the formaldehyde in the air
sample, (see Sec. 7.3.2)
7.0 PROCEDURE
7.1 Calibration
7.1.1 Calibration of the Air Pump
7.1.1.1 Using the guidelines in the instrument operating
manual, adjust the stroke and electric motor speed to produce a
sample airflow rate of approximately 1.0 L/min. (Ref. 5)
7.1.1.2 Using a 500 ml soap bubble flowmeter or other airflow
rate calibration device and the airflow control potentiometer, fine
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tune the airflow rate so that it is calibrated to the nearest 1.0
mL/min.
7.1.1.3 The air pump should be recalibrated every 30 days
during regular operation, whenever the pump diaphragm is replaced,
or whenever there is a change in operational airflow rate.
7.1.2 Flow Rate of the Formaldehyde Standard Peristaltic Pump
Tubing - With analyzer operating normally, connect a 1.0 ml Mohr pipet,
with 0.01 mL graduations, in line with the formaldehyde solution delivery
tube. Calculate the actual liquid flow rate, to the nearest 0.01 mL/min,
by timing small air bubbles introduced into the flow stream as they pass
through a fixed volume in the pipet. Recalculate the flow rate after
every 7 days of regular operation or after replacement of the pump tubing.
7.1.3 Colorimeter Calibration
7.1.3.1 Prepare 3 calibration standards which bracket the
working range of the instrument using the concentration range given
in Sec. 5.11. The lowest concentration should be adjusted to just
above the method detection limit. Further, the calibration curve
working range must bracket the sample analyte concentration.
7.1.3.2 Sample water for at least 10 minutes to establish a
baseline and to be sure the system is clean and interference free.
7.1.3.3 Sample each of the 3 standards for 10 to 15 minutes.
Between each calibration standard a water sample should be run for
10 minutes. The calibration standards should be analyzed starting
with the lowest concentration and ending with highest concentration.
7.1.3.4 Find the established baseline from the analysis data
and measure the height of each of the 3 calibration standard peaks.
Plot peak height versus concentration (/xg/m3 liquid standard
equivalent) and perform a linear regression analysis of the
calibration data to verify linearity.
7.1.3.5 A full 3 point calibration should be performed at
least once a week during routine operation. Analysis of a single
mid-range calibration standard must be performed as a check at the
beginning and end of any sampling period. A mid-range calibration
standard must also be analyzed once every 4 hours to verify
instrument calibration.
NOTE: It is possible to calibrate the TGM 555 analyzer using gaseous
formaldehyde standards, however, the procedures required to produce
accurate, dynamic, low-concentration standard mixtures in air are non-
routine. The techniques developed for use in evaluating this procedure
employ a 3-stage dynamic gas dilution system coupled with a constant-rate
vapor generation assembly containing a trioxane permeation tube (VICI
Medtronics Dynacal permeation device or equivalent) that is maintained at
55°C (See Appendix A). Trioxane vapor is converted stoichiometrically to
8520 - 8 Revision 0
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formaldehyde vapor using a special
converter assembly. (Ref. 4)
high-temperature (160°C) catalytic
7.2 Determination of Volume to be Sampled - In this method, the volume
of ambient air to be sampled is dependent on the fact that the air must be mixed
at a certain rate with water. This aqueous solution is then mixed at a certain
rate with a reactant to form the chromophore which gives an immediate (after a
short time delay) and continuous determination of the target analyte. The
sampling rate set for this equipment has been established at about 1.0 L/min.
See Sec. 8.4 and Table 3 for information regarding tests to determine the bias
of the monitoring system.
7,3 Calculations
7.3.1 Determination of Equivalent Concentration,
jug/m3, of Liquid Formaldehyde Standard Solution.
in units of
(Cstd)(LFR)
(AFR)(AE)
Where:
"std
LFR
AFR
AE
Concentration of standard solution in
Liquid flow rate calculated in Sec. 7.1.2 in mL/min
Air flow rate calculated in Sec. 7.1.1 in m3/nnn
Absorption efficiency of the air scrubber coil
(AE = 100% or 1.0 for this procedure)
7.3.2 Determination of Formaldehyde Concentration, X, in Air
Samples.
X =
Where:
Y .
b
m
(Y - b)
m
= Height of peak or plateau from analysis data.
= The Y intercept of the calibration line from linear
regression.
= The slope of the calibration line from linear
regression.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
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8.2 Method Blank - The analysis system is zeroed using a regulated source
of zero grade air. This "zero air supply" allows the analyzer to establish a
baseline response, help zero the colorimeter, and calibrate the system.
8.3 Calibration Blank - Analyze a blank between each calibration standard.
8.4 Precision and Bias
8.4.1 The precision of replicate liquid equivalent standard
analyses should be ±5 percent, for the determination made within a 24 hour
period.
8.4.2 For day to day standard determinations, the precision should
be better than ±10 percent of the calculated known value for the
standards. (Ref. 4)
8.4.3 If the analyzer performance does not exceed or meet these
requirements, instrument recalibration procedures are required.
9.0 METHOD PERFORMANCE
9.1 Method Sensitivity
9.1.1 Accurate determination of the sensitivity of this procedure
is complicated because of the difficulty involved in reproducing dynamic
test mixtures at less than 12 /xg/m . (Ref. 4) Very low ambient
concentrations, on the order of 6 jug/m3, produce a measurable response
(about 10% of the full scale) on the analyzer; however, the precision of
such measurements has not been determined.
9.1.2 As the volume of the air sampled is increased, the
sensitivity of the method increases proportionally. The standard
operating airflow rate for the analyzer is between 0.5 L/min and
1.0 L/min.
9.1.3 This method has demonstrated excellent sensitivity in
applications involving the monitoring of domestic ambient air
environments. Data in Table 1 are from a test involving low-concentration
formaldehyde measurements taken before and after changes in the heating,
ventilation, and air conditioning (HVAC) system operation.
9.2 Typical data obtained from a multipoint calibration of the monitor,
using a 1.0 volt full-scale output range on the recorder and an airflow rate of
1.0 L/min on the analyzer, is shown in Table 2. (Ref. 4)
9.3 Determination of Bias
9.3.1 Tests performed to measure the bias of this procedure were
conducted with a laboratory gas dilution system capable of generating
dynamic test mixtures of formaldehyde vapor in air, at flow rates up to
5.0 L/min and concentrations in the range from 12 to 500 jug/m3. (Ref. 4)
The output of the heated permeation tube was determined, at weekly
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intervals, by accurate weighing on a calibrated analytical balance. See
Table 3 for test results.
9.3.2 The results of the tests listed in Table 3 determined that
the accuracy of the permeation tube output was on the order of ±3.5
percent and the accuracy of the several flow calibration devices used to
generate the test mixtures is on the order of ±5 percent. These accuracy
results do not indicate that the modified CEA analyzer procedure displays
any significant bias.
10.0 REFERENCES
1. Miksch, R.R., Anthon, D.W., Fanning, L.Z., Hallowell, C.D., Revzan, K.,
and Glanville, J., "Modified Pararosaniline Method for the Determination
of Formaldehyde in Air," Analytical Chemistry, Vol. 53, 1981, pp. 2118-
2123.
2. Lyles, G.R., Bowling, F.B., and Blanchard, V.J., "Quantitative
Determination of Formaldehyde in the Parts Per Hundred Million
Concentration Level," Journal of the Air Pollution Control Association,
Vol. 15, 1965, pp. 106-108.
3. West, P.M. and Gaeke, G.C., "Fixation of Sulfur Dioxide as
Disulfitomercurate (II) and Subsequent Colorimetric Estimation,"
Analytical Chemistry, Vol. 28, 1956, pp. 1816-1819.
4. Fortune, C.R., Daughtrey, E.H., and McClenny, M.A., "Development of a
Portable Continuous Monitor for trace levels of Formaldehyde in Air," in
Proceedings of the Annual Meeting of the Air and Waste Management
Association, June 25-30, 1989, Anaheim, CA, Paper No. 89.81.2.
5. Instrument Operational Manual, CEA Instruments, Inc., 16 Chestnut St.,
P.O.Box 303, Emerson, N.J. 07630, Model TGM 555-FO.
6. "Standard Test Method for Continuous Measurement of Formaldehyde in Air,"
Annual Book of ASTM Standards, D5221.
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TABLE 1.
EXAMPLE OF AMBIENT HCHO MEASUREMENTS OF A PRIVATE RESIDENCE8
Analyzer Location
Kitchen/dining area
Kitchen/dining area
Kitchen/dining area
Bedroom area
Bedroom area
Time of Day
14:10
14:46
15:02
22:34
23:34
HCHO (fj.g/m3)
42.9
52.0
33.9
56.9
29.8
Conditions
HVAC on/house closed
HVAC off/house closed
HVAC off/house open
HVAC off/house closed
HVAC off/house open
a Measurements performed on 2-15-89.
TABLE 2.
MODIFIED CEA MONITOR MULTIPOINT CALIBRATION DATA"-"
(For formaldehyde liquid standard concentrations versus CEA monitor readings)
HCHO Liquid Std.
Equivalent (^g/m3)
30.9
61.7
92.5
123.4
CEA Monitor Reading
(% of full scale)
26.1
51.7
77.2
101.3
Recorder Response
(mm)
65
129
193
253
" Analysis performed using a 1.0 volt full-scale output range on the
recorder and an air flow rate of 1.0 L/min.
b Calculated regression coefficients are slope = 0.8144, y-intercept = 1.26,
and correlation coefficient (r) = 0.99989.
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TABLE 3.
EXAMPLE OF CEA MONITOR DILUTION SYSTEM MEASUREMENTS8
Dilution
Primary
2
2
3
3
3
3
.000
.775
.800
.800
.800
.800
System Flow Rates (L/min)
Secondary
1
2
3
0
0
0
.000
.000
.000
Gas Removal
0
0
0
1.000
2.000
3.000
Total
2
2
4
4
4
4
.200
.975
.000
.000
.000
.000
HCHO
Calculated
124
92
68
51
34
17
.7
.3
.6
.4
.3
.2
Cone.
(uq/m3)
Measured %
124
87
65
48
30
10
.5
.9
.1
.2
.0
.9
Recovery
99.8
95.2
94.9
93.8
87.5
63.4
8 Measurements were performed on 2-7-89 with the standard delivery system
operating at an output rate of 274 ±10 ng/min formaldehyde.
b Regression analysis of these measurement data taken at 6 air
concentrations resulted in: slope = 1.042 and correlation coefficient (r)
= 0.99962. The data are presented graphically in Figure 2.
8520 - 13
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FIGURE 1.
FLOW DIAGRAM OF FORMALDEHYDE ANALYZER
(A) SAMPLE CELL © AIR SCRUBBER
(£) REFERENCE CELL © LIQUID/AIR SEPARATOR
© COLORIMETER (£) GLASS DOUBLE MIXING TEE
DRAIN TEE (7) SINGLE 2S-TURN MIXING COIL
(tj GLASS DEBUBBLER (7) DOUBLE 11-TURN MIXING COIL
(7) PLASTIC TEE
DOUBLE « TURN TIME DELAY COIL
SINGLE 42-TURN TIME DELAY CON.
-fr-
8520 - 14
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FIGURE 2.
CEA MONITOR DILUTION MANIFOLD MEASUREMENTS
105
90
_: 75
o
3
5
60
45
IS
LINEAR REGRESSION DATA (y • m« » b)
N.6
b. 5 180
mm 1 042
r•0 999S2
I
I
I
IS 30 45 60 75 90
HCHO CONCENTNATION (CALCULATCO). ppbv
105
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METHOD 8520
CONTINUOUS MEASUREMENT OF FORMALDEHYDE IN AMBIENT AIR
7.1.1.1 Adjust air
pump to produce a
sample airflow rate of
approximately
1.0 L/min.
7.1.1.2 - 7.1.1.3
Calibrate airpump
to the nearest
1 .0 mL/min. (The pump
should be re-calibrated
as needed.)
7.1.2 Calculate the
actual liquid flow rate
of the formaldehyde
standard peristaltic
pump tubing.
7.1 .3 Prepare
calibration standards
and perform calibration
of the colonmter.
7.2 Determine volume
at ambient air to
be sampled.
Perform air sampling
as specified in Section
6.2, Routine Operating
Procedure.
7.3.1 Calculate the
equivalent concen-
tration (CEQ ) of liquid
from formaldehyde
standard solution.
7.3.2 Calculate the
formaldehyde con-
centration in the
air samples.
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APPENDIX A
FORMALDEHYDE VAPOR STANDARD GENERATOR
A.I A laboratory apparatus may be constructed to produce a continuous,
steady-state mixture of formaldehyde vapor at part-per-billion-by-volume (ppbv)
concentrations. The apparatus consists of a clean air generator system, a gas
dilution system, and a formaldehyde vapor generator. These three components are
shown in Figures 1A, 2A, and 3A, respectively.
A.2 Primary Dilution Air Supply
A.2.1 An in-house compressed air supply is connected to a dual
silica-gel cartridge assembly for moisture removal (Figure 1A). Use of
the appropriate valves and tube fittings permits easy replenishment of
spent cartridges while maintaining continuous operation of the system.
A.2.2 The flow of dry air to the dilution manifold is regulated by
a mass flow controller (Tylan Model FC 260) calibrated for a 0-5.0 L/min
operating range.
A.2.3 The dry air is purified with a 400 cm3 capacity gas purifier
(Alltech Assoc.). The front half of the purifier is filled with activated
charcoal to remove organic vapors, and the back half is filled with 13X
molecular sieve to remove any remaining moisture and formaldehyde vapor.
A.2.4 The primary dilution air from this system enters the three-
stage gas dilution manifold (Figure 2A) at point (A).
A.3 Three-Stage Dynamic Gas Dilution Manifold
A.3.1 Construction of Manifold
A.3.1.1 The manifold may be constructed by connecting three
sections made from high-purity quartz, each of 10 mm ID and about 50
cm length. All connection ports are 1/4 in. OD to permit the use of
standard compression fittings (PTFE Teflon) to assemble the
components.
A.3.1.2 All tubing used throughout the system is FEP Teflon
(Cole-Parmer). The 1/4 in. union fitting used to connect the first
and second stages of the manifold may be modified by snugly fitting
a short length of 2 mm ID Teflon tubing into the union. This
restriction decreases the size of the air passage by about one-half,
which increases the air velocity significantly at this point to
minimize any potential back-flushing of the secondary dilution air.
The manifold assembly was wrapped over its entire length with a
resistive wire connected to a variable transformer (output setting
= 10%).
A.3.1.3 A K-type thermocouple may be attached to the surface
of the third stage of the manifold, and the entire assembly is
wrapped with a 1/2 in. thick foam insulation material. The
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temperature of the manifold should remain constant at 30°C, varying
by no more than ±1"C with changes in the room temperature.
A.3.1.4 The exit end of the third-stage distribution manifold
is connected to a 4.0 L/min rotameter to permit easy visual
determination of the approximate exit flow velocity. The excess gas
flow is continuously vented to the fume hood through the exhaust
manifold.
A.3.2 Gas Flow Rates - Gas flow rates into and out of the manifold
are regulated by four calibrated mass flow controllers [Tylan Model FC
260) selected in various flow ranges up to 5.0 L-min to suit each
requirement. Flow rates are selected and monitored by using a Tylan Model
RO-28 readout/control unit. Four 120 cm3 capacity gas purifiers (Alltech
Assoc.) and four on/off valves (Swagelock, 316 ss) are positioned on
either side of the flow controllers.
A.3.3 Formaldehyde Dilution Scheme - The following serial dilution
scheme is versatile and responds quickly to adjustments of the flow
systems; that is, equilibration is achieved within 1-3 min after
adjustments are made. More importantly, this design permits the
generation of a tenfold-more-dilute dynamic mixture than a single-stage
method using the same volume of gas.
A.3.3.1 A continuous stream of formaldehyde in N2 (200 cm3/cm)
enters the first stage of the manifold at point (B)(Figure 2A),
where it is mixed with the primary dilution air (up to 5.0 L/min).
A.3.3.2 Near the end of the first stage, up to 90% of the
primary gas mixture may be removed by adjusting the flow controller
to the desired second stage of the dilution manifold, secondary
dilution of the gas mixture with zero air (< 5.0 L/min) may be
performed in an analogous manner.
A.3.3.3 The third stage of the dilution manifold is designed
with multiple sampling ports, where the fully diluted sample gas
stream (generally flowing at 3.0-5.0 L/min) is available for test
purposes. The second mass flow controller/purifier assembly
connected to the house vacuum line is available for use with an
optional extractive sampling method at flow rates less than or equal
to 2.0 L/min.
A.3.4 The dilution manifold is connected to the formaldehyde vapor
generator and gas dilution system shown in Figure 3A.
A.4 Low-Concentration Formaldehyde Vapor Generator/Gas Dilution System
A.4.1 The formaldehyde vapor generator (Figure 3A) may be
fabricated mostly from available spare components and some custom-made
glass, quartz, and machined Teflon parts.
A.4.2 A solid aluminum heater block (10.7 cm diameter x 25.4 cm
height) is drilled to house the permeation tube holder. An adjacent
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thermowell is drilled to house a K-type thermocouple and a ceramic
insulated RDT sensor for the temperature controller. The aluminum block
is wrapped with a heavy-duty (silicone rubber-coated) heat tape, several
coils of 1/8 in. copper tubing (to preheat the N2 carrier gas), and a
blanket of foam insulation. This assembly is mounted inside an insulated
metal box (29 cm length x 30 cm 18 cm width x 30 cm height). A small
opening is cut in the hinged lid to permit removal of the permeation tube
holder without opening the box.
A.4.3 The temperature controller (RFL Industries) is mounted to the
wall inside the insulated compartment to decrease the variability in the
set point temperature fluctuations. The externally adjustable controller
is set to maintain a set point temperature of 55°C (as measured by the
heater block thermocouple).
A.5 Permeation Tube and Holder - The permeation tube holder was
constructed from a borosilicate glass vial (2.5 cm ID x 12.5 cm height) with a
threaded opening. A threaded closure with a Viton 0-ring seal was custom-
machined from a solid piece of PTFE Teflon. The cap has two openings, each
fitted with a 1/8 in. tube fitting. Preheated N2 enters the holder, passes
through a 10.0 cm length of 1/8 in. OD FEP tubing, and sweeps the trioxane vapor
from the permeating tubeholder into the phosphoric acid converter. A Dynacal
permeation tube (5.0 cm active length) containing trioxane (mp 64°C) was
purchased from Vici Metronics (part No. 100-050-3520). The nominal permeation
rate for this device is 680 ng/min cm (±15%) at 90°C. Periodic gravimetric
determinations of weight loss over a period of 36 days with this system resulted
in a measured rate of about 56 ng/min cm at 55°C.
A.6 Converter
A.6.1 The high-temperature converter for depolymerizing the
trioxane vapor may be constructed from an 8.0 cm section of 1.0 cm ID
quartz tubing tapered at each end to 1/4 in OD for tubing connections. A
7 mm diameter indention 2 mm deep was made at the center of the tube for
attaching a temperature sensor and thermocouple wire.
A.6.2 The converter catalyst is prepared by saturating 14-mesh
silicon carbide boiling chips (GFS Chemicals) with 85% phosphoric acid
(Fisher Sci. Co.); saturation was ensured by heating for 1 hour and
soaking overnight. The quartz tube is packed tightly with catalyst
material, and small portions of quartz wool (Refrasil) were used to plug
each end.
A.6.3 The converter tube is wrapped with a nickel-chromium heater
wire, a layer of high-temperature fiberglass tape, and a double layer of
fiberglass insulation wrap. The leads of the heater wire were connected
to a variable transformer (output setting = 16%) that is wired to a
temperature controller (Omega Engineering, Model 6102-J) The ceramic
insulated J-type sensor from the controller and an exposed-junction K-type
thermocouple are attached to the converter tube at the same point by using
a silicon sealant. The temperature controller set point is adjusted to
maintain the catalyst temperature at 160°C.
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A.6.4 The outlet end of the converter assembly is connected by a
short length of 1/8 in. tubing to the first stage of the dilution
manifold; see Figure 2A, point (B).
A.7 Operation - The formaldehyde vapor generator/gas dilution system is
maintained in continuous operation. The primary dilution air is normally set at
3.8 L/min and the N2 carrier gas at 200 cm3/min for a continuous total flow of
4.0 L/min. An Omega Engineering digital thermocouple meter with a 10-position
selector switch (Model DSS-650) is used to monitor the three elevated temperature
zones in the system, the CEA monitor analytical module, and the ambient air
temperature. A matched set of five exposed-junction K-type thermocouple wires
with PFA Teflon insulation (Omega Engineering) are used.
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FIGURE 1A
PRIMARY DILUTION AIR SUPPLY
MASS FLOW
CONTROL PANEL
fTTTTm
o o o o
ACTIVATED
CHARCOAL
13X -v
MOLECULAR VI
SIEVE
PURIFIER
DILUTION
MAMFOLD
PRIMARY
DILUTION
AIR
QUICK
CONNEa
FITTINGS
"\J
SILICAGEL
DRYERS
ON/OFF
VALVE
THREE-WAY
VALVE •—}
/
PRESSURE
REGULATOR
COMPRESSED
AIR SUPPLY
8520 - 21
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TC®-AMBIENT AIR
TC® - DILUTION MANIFOLD
FIGURE 2A
THREE-STAGE DYNAMIC GAS DILUTION MANIFOLD
TO EXHAUST HOOD
TAMETER /
(0-4.0 L/min) ;
u £"~ijTOM~~/~>'
TO OPTIONAL SAMPLING METHOD
FROM
FORMALDEHYDE
GENERATOR
8520 - 22
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FIGURE 3A
LOW-CONCENTRATION FORMALDEHYDE VAPOR GENERATOR/GAS DILUTION SYSTEM
BH TO DILUTION MANIFOLD
(160 'Q
(OPTIONAL SAMPLING METHOD
TEMPERATURE
NTROLLER
(55 X)
»SP
Hill
o o o o
MASS FLOW
CONTROL PANEL
ZERO
(HCHO)}
PERMEATION
TUBE
HOUSE
VACUUM
OP
ZERO
AIR
HEATER
BLOCK
110 VAC
(INSULATED
BOX
TC® - PERM. TUBE HEATER
TC® - H3PO« CONVERTER
8520 - 23
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4.4 IMMUNOASSAY METHODS
The following methods are included in this section:
Method 4000: Immunoassay
Method 4010A: Screening for Pentachlorophenol by Immunoassay
Method 4015: Screening for 2,4-Dichlorophenoxyacetic Acid by
Immunoassay
Method 4020: Screening for Polychlorinated Biphenyls by
Immunoassay
Method 4030: Soil Screening for Petroleum Hydrocarbons by
Immunoassay
Method 4035: Soil Screening for Polynuclear Aromatic
Hydrocarbons (PAHs) by Immunoassay
Method 4040: Soil Screening for Toxaphene by Immunoassay
Method 4041: Soil Screening for Chlordane by Immunoassay
Method 4042: Soil Screening for DDT by Immunoassay
Method 4050: TNT Explosives in Water and Soils by Immunoassay
Method 4051: Hexahydro-l,3,5-trinitro-l,3,5-triazine (RDX) in
Soil and Water by Immunoassay
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METHOD 4000
IMMUNOASSAY
1.0 SCOPE AND APPLICATION
1.1 Immunoassay is an analytical technique useful for the separation,
detection and quantitation of both organic and inorganic analytes in diverse
environmental and waste matrices. Immunoassay methods are used to produce two
types of quantitative results: 1) range-finding or screening results indicative
of compliance with an action level, and 2) assay values.
1.2 Commercially-avail able testing products present immunoassay protocols
that are rapid, simple and portable. These products can be used effectively in
both laboratory and field settings, and require limited training. These test
products substantially increase the number of data points that can be generated
within a given time period, and permit an operator to analyze a number of
samples simultaneously, within a relatively short period of time. Results are
available immediately upon completion of the test, and can assist in the on-site
management of personnel and equipment, as well as the data management activities
of the laboratory.
1.2.1 A list of approved immunoassay testing products is available
from the USEPA Office of Solid Waste Methods Section.
1.3 Section 11.0 provides a glossary of basic immunoassay terms.
1.3.1 The glossary is not intended to be comprehensive, but to
provide basic definitions that will assist in understanding product
inserts and publications relating to immunoassay technology.
1.3.2 The performance of test products will vary from manufacturer
to manufacturer. The performance claims and limitations of each test
product will be provided in the package insert. The package insert of
each test product purchased should be read to determine if the performance
is acceptable for a given application.
2.0 SUMMARY OF METHOD
2.1 The immunoassay test products available will often vary in both format
and chemistry. The characteristics of a specific product are described in the
package insert provided by the manufacturer. This summary is, therefore, general
in scope, and is intended to provide a general description of the more common
elements of these methods.
Immunoassay test products use an antibody molecule to detect and quantitate
a substance in a test sample. These testing products combine the specific
binding characteristics of an antibody molecule with a detection chemistry that
produces a detectable response used for interpretation. In general, antibody
molecules specific for the method's intended target are provided at a predefined
concentration. A reporter (i.e., signal generating) reagent, composed of the
4000 - 1 Revision 0
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target compound conjugated to a signal producing compound or molecule (e.g.,
enzymes, chromophores, fluorophores, luminescent compounds, etc.), is also
provided. The concentration, affinity, and specificity of the products's
antibody influences performance, as does the chemistry of the reporter reagent.
The reporter reagent and antibody molecules of a given product are binding
partners, and complex in solution. The addition of a positive sample containing
the target substance to this solution results in a competitive binding reaction
for the antibody sites. The antibody concentration, and therefore binding
capacity, is limited to prevent the simultaneous binding of both the reporter and
sample molecules. The concentration of reporter reagent that can bind to the
antibody is inversely proportional to the concentration of substance in the test
sample. Immunoassay methods may be heterogeneous (i.e., requiring a wash or
separation step), or homogeneous (i.e., not requiring a separation step). In
commonly available heterogeneous testing products, the antibody is immobilized
to a solid support such as a disposable test tube, and the bound reporter reagent
will be retained after removing the unbound contents of the tube by washing.
Therefore, a negative sample results in the retention of more reporter molecules
than a positive sample. The analysis of a standard containing a known
concentration results in the immobilization of a proportional concentration of
reporter reagent. A positive sample (i.e., containing a higher concentration
than the standard) results in the immobilization of fewer reporter molecules than
the standard, and a negative sample (i.e., containing less that the standard)
will immobilize more.
2.2 A chemistry of the detection of the immobilized reporter is used for
interpretation of results. The reporter molecule may be a conjugate of the
target molecule and a directly detectable chromophore, fluorophore, or other
specie, or conjugated to an enzyme that will act upon a substrate to produce the
detectable response. Immunoassay testing products have a quantitative basis, and
will produce a signal that is dependant on the concentration of analyte present
in the sample. For environmental immunoassay methods, the signal produced is
exponentially related to the concentration of the compounds present. Many
immunoassay methods use enzymes to develop chromogenic response, and are termed
enzyme immunoassays. Assays that generate a chromogenic response are analyzed
photometrically, and use the principles of Beer's Law (Absorbance = Extinction
Coefficient x Concentration x Path Length) to determine the concentration of
analyte in a sample.
Immunoassay methods can provide quantitative data when configured with a series
of reference standards that are analyzed and used to construct a standard curve.
The signal generated from the analysis of a test sample is used to determine
concentration by interpolation from the standard curve. Alternatively, these
testing products can be configured to determine if a sample is positive or
negative relative to a single standard.
Individual immunoassay testing products are reviewed and accepted by the EPA-OSW
for the detection of sample analytes in specified matrices. A variety of testing
products, produced by several different developers, may be available for the same
compound(s) and matrices. Each of theses methods have been formulated using
independently developed reagents that may result in significantly different
performance characteristics and limitations.
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The performance of the immunoassay testing products ultimately relates to the
characteristics of the antibody, reporter molecule, and sample processing
chemistry. The dose-response characteristics of a method, the position of the
standard relative to the claimed action level, and the stated cross-reactivity
characteristics of the selected test product, provide relevant information
regarding the performance and recognition profile of the selected test product.
The precision, and ultimately the sensitivity of an immunoassay method, is a
function of the signal-to-noise characteristics of its dose-response curve, and
its operational consistency. Methods having a high slope and low non-specific
signal generation produce the most sensitive and precise methods. Signal
imprecision applied to a dose-response curve having a shallow slope exhibits
proportionally greater imprecision in the calculated concentration than would a
method having a steeper slope. In an action level testing product, this would
cause the reference standard to be positioned further from the action level,
increasing the incidence of false positive results. Similarly, a method having
less non-specific signal generation (higher signal-to-noise ratio) will be more
sensitive and precise when other characteristics (/.e., dose-response slope) are
held constant.
Immunoassay methods are used to detect contamination at a specific concentration
below the claimed detection level for the test product. For example, an
immunoassay used to detect PCB contamination in soil at 1 ppm will include a
reference preparation containing less than 1 ppm. The reference preparation
concentration is positioned to minimize the incidence of false negative results
at the claimed detection level. For remediation and monitoring applications,
where action levels of interest are defined, immunoassay methods should exhibit
a negligible incidence of false negative results, and minimal false positives.
For a single point action level test, the concentration of analyte relative to
the action level is selected by the developer, and is influenced by the precision
(i.e., intra-assay, inter-person, inter-lot, inter-day, etc.), sample matrix
interferences and other performance characteristics and limitations of the basic
method. The concentration of analyte in the reference materials should be less
than, but close to, the claimed action level. The concentration selected for the
standard defines the concentration that will produce a 50% incidence of false
positive results by the test product. While this issue is one representing
limited liability to the operator, it is a practical issue that often requires
attention. An immunoassay method for the detection of 1 ppm of PCB using a
standard containing 0.8 ppm of PCB will experience a 50% false positive incidence
in samples containing 0.8 ppm of PCB, and some incidence of false positive
results in a sample containing between 0.8 and 1 ppm. A similar immunoassay that
uses a standard containing 0.4 ppm will experience a 50% false positive incidence
in samples containing 0.4 ppm of PCB, and some incidence of false positive
results in a sample containing between 0.4 and 1 ppm. The closer the standard
concentration is to the action level, the better the overall performance.
2.3 Cross reactivity characteristics illustrate the specificity of the
underlying immunochemistry. The antibody molecules used by a test product bind
to a target compound and then participate in the process of generating the signal
used for interpretation. Antibody molecules bind by conformational
complimentarity. These molecules can be exquisitely specific, and can
differentiate subtle differences in the structure of a compound. The binding
4000 - 3 Revision 0
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characteristics of reagents in different test products can vary, and influence
the recognition profile and incidence of false results obtained by the method.
Immunoassay methods should detect the target analytes claimed by the test product
and exhibit limited recognition for compounds and substances not specified.
3.0 INTERFERENCES
3.1 Non-target analytes may bind with the antibody present, producing a
false-positive result. These non-target analytes may be similar to the target
analytes, or they may be chemically dissimilar co-contaminants. During
evaluation of each test product for RCRA testing applications, studies were
conducted to determine these "cross-reactive" constituents. At a minimum, these
studies evaluated the response of the test product to all other similar RCRA
analytes in that analyte class, as well as for selected lists of non-RCRA
analytes. This testing scheme is designed to ensure that all other similar RCRA
analytes and likely co-contaminants are evaluated during cross-reactivity
testing. The results of these studies are presented in each method in tabular
form, providing separate data sets for each test product evaluated.
3.2 Interference in the binding of an antibody to its target compound,
or reporter molecule reagent, may occur when testing sample matrices with
confounding contaminants or circumstances (e.g., oil, pH, temperature, some
solvents). Immunoassay products contain sample processing technology that has
been developed and validated for use with specified matrices. Interferences
incurred from the testing of incompatible matrices may prevent the testing
product from meeting its performance claims, and increase the number of false
positive or false negative results. Individual immunoassay products designate
the intended sample matrices.
3.3 Immunoassay products differ in shelf-life and storage requirements.
Test products that are operated outside of the shelf-life and storage temperature
recommendations may not provide the claimed performance.
3.4 Some test products have designated temperature ranges for operation.
When these products are used, all tests must be performed within the specified
operating temperature limits, or else false negative/positive results may exceed
performance claims.
4.0 APPARATUS AND MATERIALS
4.1 Each test product will specify the apparatus and materials provided,
as well as any additional apparatus and materials necessary for performance of
the test.
5.0 REAGENTS
5.1 The two basic reagents used in immunoassay analysis are the antibody
(e.g., anti-PCP) and reporter conjugate reagent (e.g., PCP molecules bound to an
enzyme).
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5.1.1 The formation of antibodies to haptenic molecules (i.e., most
environmental contaminants) is induced by the derivatization and coupling
of molecules of the target analytes to large carrier molecules such as
albumin, hemocyanin or thyroglobulin. The increased size and complexity
of the immunogen (antigen) conjugate, once injected, is sufficient to
stimulate the immune system to produce an antibody response. The
effectiveness of the immunogen in producing antibodies having the
prerequisite binding characteristics and recognition profile is influenced
by the surface density of the chemical groups on the carrier molecule, the
nature of the bridge chemistry used, the point of attachment, the
immunization protocol, •immunogen concentration, adjuvants (i.e., immune
response stimulants), and the species of the host animal.
5.1.2 An enzyme-reporter conjugate reagent is synthesized by
coupling a target analyte or derivative of a target analyte to an enzyme,
such as horseradish peroxidase. Enzymes enhance the sensitivity of the
method by action on a substrate and the production and catalytic
amplification of the detection signal. A single enzyme molecule used in
immunoassay methods will convert approximately 106 molecules of a target
analyte into a detectable product within one minute at ambient
temperature.
5.2 Each test product will specify the reagents provided, as well as any
additional reagents necessary for performance of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Testing of solid waste by immunoassay requires production of a
reproducible, particulate free leachate. It is critical that this leachate be
produced using a solvent that allows the reproducible extraction and recovery of
the target analytes, and is compatible with the antibody/enzyme conjugate of the
immunoassay system used. Buffers, detergents, and solvents, used together or in
combination, have been used effectively for extraction. Filtration of
particulate matter may be integrated into the immunoassay test, or accomplished
as a separate step within the protocol.
6.2 The immunoassay test products included in SW-846 methods will provide
explicit waste- or medium-specific directions for handling samples and extraction
of target analytes.
6.3 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 The specific procedure for each immunoassay test product is supplied
by the manufacturer in the package insert.
7.2 The recognition characteristics, sensitivity, detection ranges(s),
effective operating temperature, interferences and cross-reactivity of the test
will depend on the product being used.
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7.3 Immunoassay methods include both a sample processing and immunoassay
component. It should be noted that the immunochemical reagents and sample
processing components supplied with each product is specific to each
manufacturer. Methods available from different manufacturers for the same
compound and application may have significantly different performance
characteristics.
8.0 QUALITY CONTROL
8.1 The performance of the tests cited in the immunoassay methods in this
manual has been reviewed, and found to be consistent with the claims that are
made in the manufacturer's literature. In order to meet this performance
expectation, the analyst must:
o Follow the manufacturer's instructions for the test product being
used,
o Use test products before the specified expiration date,
o Use reagents only with the test products for which they are
designated,
o Use the test products within their specified storage temperature and
operating temperature limits.
8.2 It is important to evaluate the performance claims and limitation
provided with each testing product to determine its application to a specific
matrix and testing program.
8.3 Refer to Chapter One for standard quality control procedures.
9.0 METHOD PERFORMANCE
9.1 A false negative is defined as a negative response for a sample
containing the target analytes at or above the stated action level. False
negative rate is measured by analyzing split samples using both the test product
and a separate reference method. False negative data are provided in each method
for each test product evaluated.
9.2 A false positive is defined as a positive response for a sample that
contains analytes below the specified action level. Like false negatives, false
positive rates are measured by analyzing split samples with both the test product
and a separate reference method. False positive data are provided in each method
for each test product evaluated.
9.3 Cross-reactivity and recognition profile data are provided at the end
of each method in tabular form, providing separate data sets for each test
product evaluated. Using these data, the analyst can evaluate if contaminants
are present which are likely to produce a false negative response, and the
magnitude of that response.
9.4 For single-point tests, sensitivity data are provided demonstrating
the concentration of target analyte(s) that can be detected with greater than 95%
confidence.
4000 - 6 Revision 0
January 1995
-------
9.5 Data are provided demonstrating the bias of the testing products
accepted. These data may be from:
o serial dilution of samples (i.e., is the recovery of target analyte
a function of concentration?),
o sample recovery studies, and
o studies correlating the results of the testing product with a
reference method.
9.6 Data are provided demonstrating that the extraction efficiency of the
test being evaluated correlates with that of the referenced method.
10.0 REFERENCES
1. S.B. Friedman, "Doing Immunoassays in the Field", Chemtech, December 1992,
pp 732-737.
2. Roitt, L., Brosstoff, J., Male, M., (eds.), Immunology, J.B. Lippincott
Co., Philadelphia, Pennsylvania, 1989
3. Stites, Daniel P., Terr, Abba I., (eds.), Basic and Clinical Immunology,
Appleton and Lange, Norwalk, Conneticut, 1991
4. Odell, W.D. and Daughaday, W.H., Principles of Competitive Protein-Binding
Assays, J.B. Lippincott Co., Philadelphia, Pennsylvania, 1971
5. Ishikawa, E., Kawai, T., Miyai, K. (eds.), Enzyme Immunoassay, Igaku-Shoin,
Tokyo, Japan, 1981
6. Tijssen, P. (ed.), Practice and Theory of Enzyme Immunoassays, Volume 15,
Elsevier, NY, NY, 1985
7. Butler, John E. (ed.), Immunochemistry of Solid-Phase Immunoassay, CRC
Press, Boca Raton, Florida, 1991
8. Ngo, T.T., Lenhoff, H.M., Enzyme-Mediated Immunoassay, Plenum Press, New
York, 1985
9. 510K of the Federal Food, Drug and Cosmetics Act, Section 21, CFR 807.87
11.0 GLOSSARY OF TERMS
Antigen A molecule that induces the formation of an
antibody.
Antibody A binding protein which is produced in response
to an antigen, and which has the ability to bond
with the antigen that stimulated its production.
4000 - 7 Revision 0
January 1995
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B Lymphocyte
(B Cell)
Carrier
Competitive
Immunoassay
Cross-Reactivity
Dose-Response
Curve
ELISA
Enzyme Conjugate
Enzyme Immunoassay
A type of lymphocyte
differentiates into an
cell.
that, upon stimulation,
antibody-secreting plasma
An immunogenic substance that, when coupled to a
hapten, renders the hapten immunogenic.
An immunoassay method involving an
competitive binding reaction.
in-vitro
The relative concentration of an untargeted
substance that would produce a response
equivalent to a specified concentration of the
targeted compound. In a semi-quantitative
immunoassay, it provides an indication of the
concentration of cross-reactant that would
produce a positive response. Cross-reactivity
for individual compounds is often calculated as
the ratio of target substance concentration to
the cross-reacting substance concentration at 50%
inhibition of the immunoassay's maximum signal X
100%.
Representation of the signal generated by an
immunoassay (y axis) plotted against the
concentration of the target compound (x axis) in
a series of standards of known concentration.
When plotting a competitive immunoassay in a
rectilinear format, the dose-response will have a
hyperbolic character. When the Iog10 of
concentration is used, the plot assumes a
sigmoidal shape, and when the log of signal is
plotted against the logit transformation of
concentration, a straight line plot is produced.
Enzyme Linked Immunosorbent Assay is an enzyme
immunoassay method that uses an immobilized
reagent (e.g.,antibody adsorbed to a plastic
tube), to facilitate the separation of targeted
analytes (antibody-bound
target substances (free
using a washing step, and
generate the signal used
of results.
components) from non-
reaction components)
an enzyme conjugate to
for the interpretation
A molecule produced by the coupling of an enzyme
molecule to an immunoassay component that is
responsible for acting upon a substrate to
produce a detectable signal.
An immunoassay method that uses an enzyme
conjugate reagent to generate the signal used for
interpretation of results. The enzyme mediated
4000 - 8
Revision 0
January 1995
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False Negatives
False Positives
Hapten
Hapten-Carrier
Conjugate
Heterogeneous
Immunoassay
Methods
Homogeneous
Immunoassay
Methods
Immunoassay
Immunogen
Ligand
response may take the form of a chromogenic,
fluorogenic, chemiluminescent or potentiometric
reaction, (see Immunoassay and ELISA)
A negative interpretation of the method
containing the target analytes at or above the
detection level. Ideally, an immunoassay test
product included in an SW-846 method should
produce no false negatives. The maximum
permissible false negative rate is 5%, as
measured by analyzing split samples using both
the test product and a reference method.
A positive interpretation for a sample is defined
as a positive response for a sample that contains
analytes below the action level.
A substance that cannot directly induce an immune
response (e.g., antibody production), but can
bind to the products of an immune response (e.g.,
antibody) when that response is induced by an
alternate mechanism. Chemical contaminants of
the environment are haptens.
The coupling of a non-immunogenic molecule (e.g.,
targeted analyte) to an immunogenic substance
(e.g., bovine serum albumin, keyhole limpet
hemocyanin) for the purpose of stimulating an
immune response.
Immunoassay methods that include steps for the
separation of substances that become bound to the
antibody from those that remain free in solution.
Immunoassay methods that do not require the
separation of bound and free substances, but that
utilize antibody molecules that can bind and
directly modulate the signal produced by the
reporter molecule (e.g., enzyme conjugate).
An analytical technique that uses an antibody
molecule as a binding agent in the detection and
quantitation of substances in a sample, (see
Enzyme Immunoassay and ELISA)
A substance having a minimum size and complexity,
and that is sufficiently foreign to a genetically
competent host to stimulate an immune response.
The molecule, ion or group
with another molecule.
that forms a complex
4000 - 9
Revision 0
January 1995
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Lymphocytes One of the five classes of white blood cells
found in the circulatory system of vertebrates.
A mononuclear cell 7-12 pm in diameter containing
a nucleus with densely packed chromatin and a
small rim of cytoplasm.
Monocl onal Identical copies of antibody molecules that have
Antibodies a common set of binding characteristics.
Polyclonal A group of antibody molecules that differ in
Antibodies amino acid composition and sequence, and that
exhibit binding characteristics. Polyclonal
antibodies are produced from a simulation of
multiple clones of lymphocytes.
4000 - 10 Revision 0
January 1995
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METHOD 4010A
SCREENING FOR PENTACHLOROPHENOL BY IMMUNOASSAY
1.0 SCOPE AND APPLICATION
1.1 Method 4010 is a procedure for screening solids such as soils,
sludges, and aqueous media such as waste water and leachates for
pentachlorophenol (PCP) (CAS Registry 87-86-5).
1.2 Method 4010 is recommended for screening samples to determine whether
PCP is likely to be present at defined concentrations (i.e., kits are available
which give positive results at 0.005 mg/L for aqueous samples, and at 0.5, 10 or
100 mg/kg in solid samples). Method 4010 provides an estimate for the
concentration of PCP by comparison with a standard.
1.3 Using the test kits from which this method was developed, 95% of
aqueous samples containing 2 ppb or less of PCP will produce a negative result
in the 5 ppb configuration. Also, 95% of soil samples containing 125 ppb or less
of PCP will produce a negative result in the 5000 ppb test configuration.
1.4 In cases where the exact concentration of PCP is required, additional
techniques (i.e., gas chromatography (Method 8040) or gas chromatography/mass
spectrometry (Method 8270)) should be used.
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 Test kits are commercially available for this method. The
manufacturer's directions should be followed.
2.2 In general, the method is performed using a water sample or an extract
of a water sample. The sample/extract and an enzyme conjugate reagent are added
to immobilized antibody. The enzyme conjugate "competes" with PCP present in the
sample for binding to immobilized anti-PCP antibody. The test is interpreted by
comparing the response produced by testing a sample to the response produced by
testing standard(s) simultaneously.
3.0 INTERFERENCES
3.1 Compounds that are chemically similar may cause a positive test (false
positive) for PCP. The test kits used in preparation of this method were
evaluated for interferences. Tables 1A and IB provide the concentration of
compounds which will give a false positive test at the indicated concentration.
3.2 Other compounds have been tested for cross reactivity for PCP and have
been demonstrated not to interfere with the specific kits tested. Consult the
information provided by the manufacturer of the kit used for additional
4010A-1 Revision 1
January 1995
-------
information regarding cross reactivity with other compounds.
3.3 Storage and use temperatures may modify the method performance. Follow
the manufacturer's directions for storage and use.
4.0 APPARATUS AND MATERIALS
4.1 Immunoassay test kit: PENTA RISc™ (EnSys, Inc.), EnviroGard™ PCP in
Soil (Millipore, Inc.), or equivalent. Each commercially available test kit will
supply or specify the apparatus and materials necessary for successful completion
of the test.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Soil samples may be contaminated, and should therefore be considered
hazardous and handled accordingly.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance specifications indicated
in Tables 2-10.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for the test kit being used for
quality control procedures specific to the test kit used. Additionally, guidance
provided in Method 4000 and Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.3 Do not use test kits past their expiration date.
8.4 Do not use tubes or reagents designated for use with other test kits.
8.5 Use the test kits within their specified storage temperature and
operating temperature limits.
4010A-2 Revision 1
January 1995
-------
8.6 Method 4010 is intended for field or laboratory use. The appropriate
level of quality assurance should accompany the application of this method to
document data quality.
9.0 METHOD PERFORMANCE
9.1 This method has been applied to a series of groundwater, process
water, and wastewater samples from industries which use PCP, and the results
compared with GC/MS determination of PCP (Method 8270). These results are
provided in Table 2. These results represent determinations by two laboratories
using the PENTA RISc™ test kit.
9.2 This method has been applied to a series of soils from industries
which use PCP and the results compared with GC/MS determination of PCP via method
8270. These results are provided in Table 3. These results represent
determinations by two laboratories using the PENTA RISc™ test kit.
9.3 Sensitivity of the EnviroGard PCP in Soil Test Kit was determined by
establishing the "noise" level expected from matrix effects encountered in
negative soil samples and determining the corresponding TPH concentration by
comparison to the analyte-specific response curve. Eight different soils which
did not contain PCP were assayed. Each of these soils was extracted in
triplicate and each extract was analyzed in three different assays. The mean and
the standard deviation of the resulting %Bo's (%Bo = [(OD8ample/ODnegatlvecontrol)xlOO])
were calculated and the sensitivity was estimated at two standard deviations
below the mean. The sensitivity for Method 4010 was determined to be 80% Bo at
a 95% confidence interval. Based on the average assay response to PCP, this
corresponds to 2 ppm PCP. These data are shown in Table 4.
9.4 The effect of water content of the soil samples on the EnviroGard™ PCP
in Soil test kit was determined by assaying three different soil samples which
had been dried and subsequently had water added to 30% (w/w). Aliquots of these
samples were then fortified with PCP. Each soil sample was assayed three times,
with and without added water, and with and without home heating oil (HHO)
fortification. It was determined that water in soil up to 30% had no detectable
effect on the method. These data are shown in Table 5.
9.5 The effect of the pH of the soil extract on the EnviroGard™ PCP in
Soil test kit was determined by adjusting the soil pH of three soil samples.
Soil samples were adjusted to pH 2 - 4 using 6N HC1 and pH 10 - 12 using 6N NaOH.
Aliquots of the pH adjusted soil samples were fortified with PCP and the
unfortified and fortified samples were extracted. These extracts were assayed
three times. It was determined that soil samples with pH ranging from 3 to 11
had no detectable effect on the performance of the method. These data are shown
in Table 6.
9.6 The bias of the EnviroGard™ PCP in Soil test kit was estimated by
fortifying three different soil samples at two different concentrations (10 and
100 ppm PCP). Each fortified sample was extracted three times and each extract
was assayed three times. Recovery for individual determinations ranged from 60%
to 125%. Average recovery for each individual extract ranged from 72% to 101%.
Overall average recovery for all samples was 86%. These data are summarized in
4010A-3 Revision 1
January 1995
-------
Table 7.
9.7 The effect of co-contamination of soil samples with oil on the
EnviroGard™ PCP in Soil test kit was investigated. Three soil samples were
adulterated with diesel oil and aliquots were fortified with PCP. The samples
were extracted and the extracts each assayed three times. It was determined that
no interference was detected in samples with up to 10% oil contamination. The
data from samples adulterated at 10% are shown in Table 8.
9.8 A field trial was conducted at a contaminated site using the
EnviroGard™ PCP in Soil test kit. Method 4010 was used to identify soil which
had been contaminated with PCP from wood treatment operations. A total of 33
samples were analyzed including 5 field duplicates. For the field duplicates,
the reference method demonstrated an average coefficient of variation of 16%.
For Method 4010 average coefficient of variation was 31%. Since Method 4010 is
not quantitative, quantitative values were estimated. These data are shown in
Table 9. At the 10 ppm cutoff, there were 0/33 (0%) false negatives and 0/33
(0%) false positives. At the 100 ppm cutoff, there was 1/33 (3%) false negatives
and 1/33 (3%) false positives. These data are shown in Table 10.
10.0 REFERENCES
1. J.P. Mapes, K.D. McKenzie, L.R. McClelland, S. Movassaghi, R.A. Reddy, R.L.
Allen, and S.B. Friedman, "Rapid, On-Site Screening Test for
Pentachlorophenol in Soil and Water - PENTA-RISc™", Ensys Inc., Research
Triangle Park, NC 27709
2. J.P. Mapes, K.D. McKenzie, L.R. McClelland, S. Movassaghi, R.A. Reddy, R.L.
Allen, and S.B. Friedman, "PENTA-RISc™ - An On-Site Immunoassay for
Pentachlorophenol in Soil", Ensys Inc., Research Triangle Park, NC 27709
3. PENTA-RISc™ Instructions for Use, Ensys Inc.
4. EnviroGard™ PCP in Soil Test Kit Guide, Millipore, Inc.
4010A-4 Revision 1
January 1995
-------
Table 1A - Cross Reactivity for POP
PENTA RISc™ Test Kit
Compound3
2,6-Dichlorophenol
2,3,4-Trichlorophenol
2,4,6-Trichlorophenol
2,4,5-Trichlorophenol
2,3,5,6-Tetrachlorophenol
Tetrachlorohydroquinone
Concentration (mg/Kg)
in Soil to Cause a
False Positive for
PCP at 0.5 mg/Kg
700
400
16
100
1.2
500
Concentration (/ng/L)
in Water to Cause a
False Positive for
PCP at 5 jiig/L
600
600
100
500
7
>1500
a Compounds assayed at 3.75 /LtM (molar equivalent of PCP at 1000 M9/L)> except
where noted.
Table IB - Cross Reactivity for PCP
EnviroGard™ PCP in Soil Test Kit
Compound
Pentachlorophenol
2,5-Dichlorophenol
2,6-Dichlorophenol
2,3,4-Trichlorophenol
2,3,5-Trichlorophenol
2,3,6-Trichlorophenol
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
Lower Limit of
Detection (mg/kg)
10
1000
1000
1000
500
500
500
500
The following compounds were tested and found to yield
negative results at 1,000 ppm:
2,3,5,6-Tetrachloronitrobenzene PCB (Aroclor 1248)
3,5-Dichlorophenol TNT
2,4-Dichlorophenol DDT
2,3-Dichlorophenol PAHs
4-Chlorophenol Chlordane
4010A-5
Revision 1
January 1995
-------
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Table 3
Comparison of PENTA RISc™ Test Kit with GC/MS
Soil Matrix
Concentration measured
by GC/MS (ppm)
1100
88
0.31
0.72
315
1.5
6.4
9
1.9
46
<1
21
3.3
4
11
18
33
54
65
74
83
1.1
14.3
<1
<1
<1
Screening
Results (ppm)
0.5
>
>
<
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AGREEMENT3
Y, FP, FN
Y
FN
Y
FN
Y
Y
Y
Y
Y
FP
Y
Y
Y
FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
4010A-8
Revision 1
January 1995
-------
Table 3
Comparison of PENTA RISc™ Test Kit with GC/MS
Soil Matrix
Concentration measured
by GC/MS (ppm)
3.9
<1
1.4
48
<1
142
108
117
56
2.5
3.5
143
nd
0.02
5
Screening
Results (ppm)
0.5
>
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Y, FP, FN
Y
Y
Y
FP
Y
Y
FN
Y
Y
Y
FP
Y
Y
Y
Y
4010A-9
Revision 1
January 1995
-------
TABLE 4
EnviroGard™ PCP in Soil Test Kit3 Sensitivity
Part 1 - Average Response with Negative Soils
Soil# Soil Type Average %Bo (n = 9)Standard Deviation ^^
SI
S2
S3
S4
S5
S6
S7
S8
LOAM
CLAY
SAND
LOAM
SAND
CLAY
LOAM/SAND
SAND/LOAM
97.6
100.1
101.4
99.4
100.2
97.4
102.6
97.5
3.0
1.4
2.8
4.9
3.1
2.7
0.3
3.6
AVERAGE 99.5 5.2
Part 2 - Average Response with Pentachlorophenol Calibrators
PCP
Concentration (ppm) Average AbsorbanceAverage %Bo
0
5
20
50
200
1.142
0.828
0.556
0.382
0.162
N/A
72.6
48.7
33.4
14.1
Part 3 - Method Sensitivity
Based on Part 1 and Part 2 Above:
Average %Bo - 2 SO = 89.2 which is equivalent to 1.6 ppm PCP
Average %Bo - 3 SD = 84.0 which is equivalent to 2.3 ppm PCP
(%Bo = IIODsample/ODnegative controllxlOOl)
4010A-10 Revision 1
January 1995
-------
TABLE 5
EFFECT OF WATER CONTENT IN SOIL SAMPLES3
Soil % Water Fortified?
Mean Std. Dev. ± 2 SD Range
SI
SI
SI
SI
S2
S2
S2
S2
S3
S3
S3
S3
0
30
0
30
0
30
0
30
0
30
0
30
No
No
Yes
Yes
No
No
Yes
Yes
No
No
Yes
Yes
104.5*
101.9
38.9
49.2
97.8
105.1
40.2
48.8
98.3
111.5
43.3
46.5
106.5
106.3
47.2
51.1
105.7
109.7
47.5
47.2
107.1
103.1
47.2
49.8
99.7
95.2
40.2
48.2
96.7
93.9
42.7
44.8
99.7
95.1
43.2
48.0
103.6
101.1
42.1
49.5
100.1
102.9
43.5
46.9
101.7
103.2
44.6
48.1
3.5
5.6
4.4
1.5
4.9
8.1
3.7
2.0
4.7
8.2
2.3
1.7
96.6
89.9
33.3
46.5
90.3
86.7
36.1
42.9
92.3
86.8
40.0
44.7
- Ill
- 112
-50.9
- 52.5
- 110
- 119
- 50.9
- 50.9
- Ill
- 120
- 49.2
- 51.5
* All values shown are %Bo [= (ODsample/ODneaatlve Control)xl00]
a EnviroGard™ PCP in Soil (Millipore, Inc.)
4010A-11
Revision 1
January 1995
-------
TABLE 6
EFFECT OF pH OF SOIL SAMPLES3
oil pH Ad.i. Fortified? Rep.
SI
SI
SI
SI
SI
SI
S2
S2
S2
S2
S2
S2
S3
S3
S3
S3
S3
S3
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
No
No
No
Yes
Yes
Yes
No
No
No
Yes
Yes
Yes
No
No
No
Yes
Yes
Yes
103.
88.
85.
52.
57.
44.
105.
104.
93.
47.
51.
43.
92.
96.
87.
55.
55.
44.
1* Rep. 2
1
7
2
7
1
6
6
4
4
8
4
3
3
6
7
2
3
3
98.6
96.9
90.9
44.8
44.6
41.6
93.9
91.3
87.7
45.1
44.4
40.7
101.8
91.9
99.8
49.5
48.3
39.3
Rep. 3
98.6
100.2
98.0
45.8
45.2
45.9
102.5
105.8
105.8
44.3
54.1
44.0
100.4
98.5
96.3
55.9
42.0
48.0
Mean Std. Dev. +2
100.1
95.3
91.3
47.8
48.9
44.0
100.7
100.5
95.6
45.7
50.0
42.7
98.2
95.7
94.6
53.6
48.5
43.9
2.6
5.9
6.4
4.3
7.0
2.2
6.1
8.0
9.3
1.8
5.0
1.8
5.2
3.4
6.2
3.5
6.7
4.4
94
83
78
39
34
39
88
84
77
42
40
39
87
88
82
46
35
35
SD Ranqe
.9 -
.5 -
.5 -
.2 -
.9 -
.6 -
.5 -
.5 -
.0 -
.1 -
.0 -
.1 -
.8 -
.9 -
.2 -
.6 -
.1 -
.1 -
105
107
104
56.4
62.9
48.4
113
117
114
49.3
60.0
46.3
109
103
107
60.6
61.9
52.7
All values shown are %Bo [= (ODsample/ODnegatlve C0ntrol)xl00]
EnviroGard™ PCP in Soil (Millipore, Inc.)
4010A-12 Revision 1
January 1995
-------
TABLE 7
Test Kit3 Bias
Soi1# Fortiflcation(ppm) Extraction^ Recovered(ppm)* % Recovery
SI 10 19 91
SI 10 29 86
SI 10 39 88
SI 100 1 84 84
SI 100 2 78 78
SI 100 3 76 76
S2 10 1 10 100
S2 10 28 76
S2 10 38 76
S2 100 1 101 101
S2 100 2 98 98
S2 100 3 88 88
S3 10 17 72
S3 10 28 76
S3 10 38 81
S3 100 1 95 95
S3 100 2 90 90
S3 100 3 87 87
Average »»»»»»»>»»»»»»»»»»»»»»»»»»» 84
Overall Average %Recovery = 86
EnviroGard™ PCP in Soil (Millipore, Inc.)
4010A-13 Revision 1
January 1995
-------
TABLE 8
Effect of Co-contamination with Diesel Oila
Soi1# Adulterated Fortified Rep.fll
SI
SI
SI
SI
S2
S2
S2
S2
S3
S3
S3
S3
NO
YES
NO
YES
NO
YES
NO
YES
NO
YES
NO
YES
Rep,
Figures are %Bo =[(ODsample/ODnegatlve control)*100]
EnviroGard™ PCP in Soil (Millipore, Inc.)
Mean
NO
NO
YES
YES
NO
NO
YES
YES
NO
NO
YES
YES
103.2*
93.4
52.7
50.9
103.1
85.4
47.8
44.6
98.9
103.8
55.2
50.4
92.5
99.4
44.8
49.7
98.3
95.1
45.1
50.8
95.4
99.7
49.5
50.6
99.8
106.2
45.8
44.6
102.3
99.9
44.3
49.0
108.1
101.4
55.9
56.7
98.5
99.7
47.8
48.4
101.2
93.5
45.7
48.1
100.8
101.6
53.6
52.6
4010A-14
Revision 1
January 1995
-------
TABLE 9
Field Duplicates3
Sample ID
059 073 074 086 087
Method 8270
Determination #1 9600 74.8 836 6.59 34.0
Determination #2 10300 78.2 1520 6.88 51.8
Average 9950 76.5 1178 6.74 42.9
Standard Deviation 495 2.4 484 0.20 12.6
% Coefficient of Variation 5.0 3.1 41 3.0 29
Immunoassay*
Determination #1 4480 79.5 604 2.4 36.0
Determination #2 3370 122 421 5.0 24.0
Average 3920 101 512 3.7 30.0
Standard Deviation 785 30.0 129 1.8 8.5
% Coefficient of Variation 20 30 25 50 28
* For the purpose of this comparison, quantitative values were calculated for
the immunoassay.
a EnviroGard™ PCP in Soil (Millipore, Inc.)
4010A-15 Revision 1
January 1995
-------
TABLE 10
Immunoassay3 Compared to Method 8270
Test Interpretation at 10 ppm PCP ^^
Sample ID Method 8270 Immunoassay Concurrence?
059 9600 >10 YES
059D 10300 >10 YES
060 1010 >10 YES
061 2740 >10 YES
063 1610 >10 YES
064 1980 >10 YES
065 1580 >10 YES
066 57.8 >10 YES
067 110 >10 YES
068 47.7 >10 YES
069 798 >10 YES
070 2890 >10 YES
071 289 >10 YES
072 326 >10 YES
073 74.8 >10 YES
073D 78.2 >10 YES
074 836 >10 YES
074D 1520 >10 YES
075 3690 >10 YES
076 4590 >10 YES
077 2040 >10 YES
078 1720 >10 YES
079 792 >10 YES
080 2550 >10 YES
081 125 >10 YES
082 2400 >10 YES
083 270 >10 YES
084 1140 >10 YES
085 57.7 >10 YES
086 6.59 <10 YES
086D 6.88 <10 YES
087 34.0 >10 YES
087D 51.8 >10 YES
EnviroGard™ PCP in Soil (Millipore, Inc.)
4010A-16 Revision 1
January 1995
-------
TABLE 10 (continued)
Immunoassay3 Compared to Method 8270
Test Interpretation at 100 ppm PCP
Sample ID Method 8270 Immunoassay Concurrence?
059 9600 >100 YES
059D 10300 >100 YES
060 1010 >100 YES
061 2740 >100 YES
063 1610 >100 YES
064 1980 >100 YES
065 1580 >100 YES
066 57.8 <100 YES
067 110 >100 YES
068 47.7 <100 YES
069 798 >100 YES
070 2890 >100 YES
071 289 >100 YES
072 326 >100 YES
073 74.8 <100 YES
073D 78.2 >100 False Positive
074 836 >100 YES
074D 1520 >100 YES
075 3690 >100 YES
076 4590 >100 YES
077 2040 >100 YES
078 1720 >100 YES
079 792 >100 YES
080 2550 >100 YES
081 125 <100 False Negative
082 2400 >100 YES
083 270 >100 YES
084 1140 >100 YES
085 57.7 <100 YES
086 6.59 <100 YES
086D 6.88 <100 YES
087 34.0 <100 YES
087D 51.8 <100 YES
4010A-17 Revision 1
January 1995
-------
METHOD 4015
SCREENING FOR 2,4-DICHLORORPHENOXYACETIC ACID BY IMMUNOASSAY
1.0 SCOPE AND APPLICATION
1.1 Method 4015 is a procedure for screening soils and aqueous matrices
to determine whether 2,4-dichlorophenoxyacetic acid (2,4-D) (CAS Registry 94-75-
7) is likely to be present at concentrations above 0.1, 0.5, 1.0 or 5.0 mg/kg in
soil, and in aqueous matrices above 10 mg/L (the toxicity characteristic
regulatory action level) and 10 /jg/L (ground water monitoring). Method 4015
provides an estimate for the concentration of 2,4-D by comparison against
standards.
1.2 Using the test kit from which this method was developed, >95% of
aqueous samples confirmed to have concentrations of 2,4-D below detection limits
will produce a negative result in the 10 ppm test configuration.
1.3 In cases where the exact concentration of 2,4-D is required,
additional techniques (i.e., gas chromatography Method 8151) should be used.
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 Test kits are commercially available for this method. The
manufacturer's directions should be followed.
2.2 In general, the method is performed using an extract of a soil
sample, or directly on an aqueous sample. Filtered extracts may be stored cold,
in the dark. An aliquot of the aqueous sample or extract and an enzyme-2,4-D
conjugate reagent are added to immobilized 2,4-D antibody. The enzyme-2,4-D
conjugate "competes" with 2,4-D present in the sample for binding to 2,4-D
antibody. The enzyme-2,4-D conjugate bound to the 2,4-D antibody then catalyzes
a colorless substrate to a colored product. The test is interpreted by comparing
the color produced by a sample to the response produced by a reference reaction.
3.0 INTERFERENCES
3.1 Compounds that are chemically similar may cause a positive test
(false positive) for 2,4-D. The data for the lower limit of detection of these
compounds are provided in Tables 1A and 1C. Consult the information provided by
the manufacturer of the kit used for additional information regarding cross
reactivity with other compounds.
4015-1 Revision 0
January 1995
-------
3.1.1 Solutions of Silvex alone, and Silvex/2,4-D mixtures, were
prepared in TCLP buffer to demonstrate the potential effect of a
structurally similar, environmentally significant cross-reactant on the
immunoassay screening results. At one-half of the action level for 2,4-D
(5ppm), 200 ppm of Silvex are required to be present to generate a false
positive response. These results are summarized in Table IB.
3.2 Storage and use temperatures may modify the method performance.
Follow the manufacturer's directions for storage and use.
4.0 APPARATUS AND MATERIALS
4.1 Immunoassay test kit: 2,4-D RaPID,, Assay kit (Ohmicron), EnviroGardIH
2,4-D in Soil (Millipore, Inc.), or equivalent. Each commercially available test
kit will supply or specify the apparatus and materials necessary for successful
completion of the test.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Soil samples may be contaminated, and should therefore be considered
hazardous and handled accordingly.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance specifications indicated
in Tables 2-9.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for the test kit being used
for quality control procedures specific to the test kit used. Additionally,
guidance provided in Method 4000 and Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
4015-2 Revision 0
January 1995
-------
8.3 Do not use test kits past their expiration date.
8.4 Do not use tubes or reagents designated for use with other test kits.
8.5 Use the test kits within their specified storage temperature and
operating temperature limits.
8.6 Method 4015 is intended for field or laboratory use. The appropriate
level of quality assurance should accompany the application of this method to
document data quality.
9.0 METHOD PERFORMANCE
9.1 Sensitivity of the EnviroGardra 2,4-D in Soil Test Kit was determined
by establishing the "noise" level expected from matrix effects encountered in
negative soil samples and determining the corresponding 2,4-D concentration by
comparison to the analyte-specific response curve. Eight different soils which
did not contain 2,4-D were assayed. Each of these soils was extracted in
triplicate and each extract was assayed in three different assays. The mean and
the standard deviation of the resulting %Bo's (%Bo = [ (ODsample/ODnegatlve Control)xl00])
were calculated and the sensitivity was estimated at two standard deviations
below the mean. The sensitivity for Method 4015 was determined to be 80% Bo at
a 95% confidence interval. Based on the average assay response to 2,4-D, this
corresponds to 0.16 ppm 2,4-D. These data are shown in Table 2.
9.2 The effect of water content of the soil samples was determined by
assaying three different soil samples which had been dried and subsequently had
water added to 30% (w/w). Aliquots of these samples were then fortified with
2,4-D. Each soil sample was assayed three times, with and without added water,
and with and without 2,4-D fortification. It was determined that water in soil
up to 30% had no detectable effect on the method. These data are shown in Table
3.
9.3 The effect of the pH of the soil extract was determined by adjusting
the soil pH of three soil samples. Soil samples were adjusted to pH 2 - 4 using
6N HC1 and pH 10 - 12 using 6N NaOH. Aliquots of the pH adjusted soil samples
were fortified with 2,4-D. Each soil sample was assayed unadjusted and with pH
adjusted to 2-4 and 10-12, both unfortified and fortified. It was determined
that soil samples with pH ranging from 3 to 11 had no detectable effect on the
performance of the method. These data are shown in Table 4.
9.4 The method bias was estimated by fortifying three different soil
samples at two different concentrations (0.3 and 2 ppm 2,4-D). Each fortified
sample was extracted three times and each extract was assayed three times.
Recovery for individual determinations ranged from 27% to 151%. Average recovery
for each individual extract ranged from 70% to 120%. Overall average recovery
for all samples was 99.7%. These data are summarized in Table 5.
9.5 The probabilities of generating false negative and false positive
4015-3 Revision 0
January 1995
-------
results at a 10 ppm action level are shown in Table 6.
9.6 The results obtained from spiking 2,4-D into TCLP leachates and other
aqueous samples are reported in Table 7. Each matrix was diluted 1:1000 and
tested by immunoassay 5 times. The results are reported as positive (+) or
negative (-). Municipal water results are based on a 52 ppb cutoff to determine
positive from negative, and were diluted 1:7.
9.7 Comparison of the results from immunoassay and GC (Method 8150)
testing of aqueous samples are presented in Table 8.
9.8 A field trial was undertaken to evaluate the ability of the
EnviroGardw 2,4-D in Soil Test Kit to identify 2,4-D contaminated soil at a
remediation site. A total of 30 soil samples were evaluated by both the
immunoassay and Method 8151. Interpretation of the results at 200 /tg/kg
resulted in 0/32 (0%) false negatives and 1/32 (3%) false positives. This
corresponds to specificity 95% and sensitivity of 100%. These data are shown in
Table 9.
10.0 REFERENCES
1. 2,4-D RaPID, Assay kit Users Guide, Ohmicron.
2. EnviroGardM 2,4-D in Soil Test Kit Guide, Millipore, Inc.
3. Lawruk, T.S., Hottenstein, C.S., Fleeker, J.R., Hall, J.C., Herzog, D.P.,
Rubio, P.M., "Quantitation of 2,4-D and Related Chlorophenoxy Herbicides by
A Magnetic Particle-Based ELISA"' 1993, (manuscript submitted for
publication).
4. Hayes, M.C., Jourdan, S.W., Lawruk, T.S., and Herzog, D.P., "Screening of
TCLP Extracts of Soil and Wastewater for 2,4-D by Immunoassay", USEPA Ninth
Annual Waste Testing and Quality Assurance Symposium, 1993.
4015-4 Revision 0
January 1995
-------
TABLE 1A
Cross-Reactivity3 of Chi orophenoxy Compounds and
Structurally Unrelated Pesticides
Compound
2,4-D
2,4-D propylene glycol ester
2,4-D ethyl ester
2,4-D isopropyl
2,4-D methyl ester
2,4-D sec-butyl ester
2,4-D butyl ester
2,4-D butoxyethyl ester
2,4,5-T methyl ester
2,4-D iso-octyl ester
2,4-D butoxy-propylene ester
2,4-DB
MCPA
2,4,5-T
Silvex methyl ester
4-Chlorophenoxyacetic acid
MCPB
Silvex (2,4,5-TP)
Dichlorophenol
Dichloroprop
Triclopyr
MCPP
Mecoprop
Pentachlorophenol
Picloram
Concentration Giving
a Positive Result
(ppm TCLP Leachate)
10
0.52
0.54
0.96
1.09
1.40
1.60
2.00
12.0
20.0
20.6
95
110
130
665
815
980
1375
2380
5000
>10,000
>10,000
>10,000
>10,000
>10,000
Percent Cross-
Reactivity
100
1900
1850
1040
917
714
625
500
86
50
49
11
9
8
1.5
1.2
1.0
0.7
0.4
0.2
<0.1
<0.1
<0.1
<0.1
<0.1
4015-5
Revision 0
January 1995
-------
TABLE 1A
Cross-Reactivity3 of Chlorophenoxy Compounds and
Structurally Unrelated Pesticides
Compound
Alachlor
Aldicarb
Aldicarb sulfate
Aldicarb sulfoxide
Atrazine
Benomyl
Butyl ate
Captan
Captofol
Carbaryl
Carbofuran
Dicamba
1,3-Dichloropropene
Dinoseb
Metolachlor
Metribuzin
Simazine
Terbufos
Thiabendazol
Concentration Giving
a Positive Result
(ppm TCLP Leachate)
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
>10,000
Percent Cross-
Reactivity
a 2,4-D RaPID^ Assay kit
TABLE IB
Cross-Reactivity3 of 2,4-D with Silvex
Si 1 vex Concentration
(ppm)
0
0.5
1.0
2.0
100
200
0
0.5
1.0
2.0
100
200
2,4-D Concentration
(ppm)
0
0
0
0
0
0
5.0
5.0
5.0
5.0
5.0
5.0
Screening Result
_
-
-
-
-
-
-
-
-
-
-
+
a 2,4-0 RaPIDtl( Assay kit
4015-6
Revision 0
January 1995
-------
TABLE 1C
CROSS REACTIVITY'
Compound
2,4-D Acid
2,4-D butyl ester
2,4-D Dichlorophenol
2,4-D isobutyl ester
2,4-D isopropyl ester
2,4-D methyl ester
2,4-DB
2,4-DB butyl ester
Dichloroprop
Diclofop
MCPA
2,4,5-T acid
Concentration Required for
Positive Interpretation (ppm)
0.2
0.025
1.5
0.2
0.2
0.1
0.2
0.9
6.0
42.5
0.8
7.0
EnviroGardIH 2,4-D in Soil Test Kit (Millipore Corporation)
4015-7
Revision 0
January 1995
-------
TABLE 2
Sensitivity of the EnviroGardw 2,4-D in Soil Test Kit
Part 1 - Average Response with Negative Soils
Soil#
SI
S2
S3
S4
S5
S6
S7
Average
Soil Type
LOAM
LOAM
SAND/ LOAM
CLAY
CLAY
LOAM/SAND
SAND
LOAM
Average %Bo
(n=9)
90.0
89.6
89.3
86.3
90.0
86.9
88.8
86.9
88.5
Standard
Deviation
1.7
2.3
2.1
1.9
2.3
2.6
2.8
2.9
6.5
Part 2 - Average Response with 2,4-D Calibrators
2,4-D Calibrator
Concentration (ppm)
0
0.1
0.5
1.0
5.0
Average
Absorbance
1.442
1.186
0.776
0.600
0.301
Average %Bo
N/A
82.2
53.8
41.7
20.9
Part 3 - Method Sensitivity
Based on Part 1 and Part 2 Above:
Average %Bo - 2 SD = 75.6 which is equivalent to 0.16 ppm 2,4-D
Average %Bo - 3 SD = 69.1 which is equivalent to 0.23 ppm 2,4-D
(%Bo = [(ODsample/ODnegatpvecontrol)xlOO])
4015-8
Revision 0
January 1995
-------
TABLE 3
Effect of Water Content of Soil Sampleson the EnviroGard™ 2,4-D in Soil Test Kit
Soil % Water Fortified? Rep. 1 Rep. 2 Rep. 3 Mean Std. Dev. ± 2 SD Range
SI
SI
SI
SI
S2
S2
S2
S2
S3
S3
S3
S3
0
30
0
30
0
30
0
30
0
30
0
30
No
No
Yes
Yes
No
No
Yes
Yes
No
No
Yes
Yes
98.
96.
61.
63.
98.
96.
47.
37.
98.
97.
41.
43.
7*
0
4
1
5
0
6
6
7
3
0
1
99.9
95.4
62.0
59.9
90.7
95.4
47.0
37.7
94.1
97.2
39.3
40.4
102.9
93.7
73.1
69.4
97.8
96.8
46.0
40.0
105.2
95.9
48.8
47.4
100.5
95.0
65.5
64.1
95.7
96.1
46.9
38.4
99.4
96.8
43.1
43.6
2
1
6
4
4
0
0
1
5
0
5
3
.2
.2
.6
.8
.3
.7
.8
.3
.6
.8
.1
.5
96.1
92.6
52.3
54.5
87.1
94.7
45.3
35.8
88.2
95.2
32.9
36.6
- 105
- 97.4
- 78.7
- 73.7
- 104
- 97.5
- 48.5
- 41.0
- Ill
- 98.4
- 53.3
- 50.6
All values shown are %Bo [= (ODsample/ODnegatlve Control)xl00]
4015-9 Revision 0
January 1995
-------
TABLE 4
Effect of pH of Soil Samples on the EnviroGardTk 2,4-D in Soil Test Kit
Soil pH Adj. Fortified? Rep. 1* Rep. 2 Rep. 3 Mean Std. Dev. ± 2 SD Range
SI
SI
SI
SI
SI
SI
S2
S2
S2
S2
S2
S2
S3
S3
S3
S3
S3
S3
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
No
No
No
Yes
Yes
Yes
No
No
No
Yes
Yes
Yes
No
No
No
Yes
Yes
Yes
95
102
96
46
50
43
94
91
89
50
56
46
82
95
86
52
55
59
.5
.8
.0
.0
.3
.7
.7
.5
.3
.9
.2
.0
.1
.2
.2
.4
92.5
105
98.3
47.5
51.9
52.4
90.6
95.8
94.2
52.6
58.1
54.2
92.0
85.1
84.4
63.6
59.5
54.3
88.7
93.1
79.4
48.6
43.7
39.1
90.8
85.9
81.0
50.2
44.3
46.4
85.4
86.9
103
49.4
66.6
54.9
92.2
100
91.5
47.4
48.5
44.8
91.9
91.1
88.3
51.1
52.9
49.1
86.5
89.0
91.2
55.1
60.4
56.2
3
6
10
1
4
6
2
5
6
1
7
4
5
5
10
7
5
2
.4
.2
.5
.3
.3
.8
.1
.0
.7
.3
.5
.4
.0
.3
.4
.5
.8
.8
85.4
87.6
70.5
44.8
39.9
31.2
87.7
81.1
74.9
48.5
37.9
40.3
76.5
78.4
70.4
40.1
48.8
50.6
- 99.0
- 112
- 113
- 50.0
- 57.1
- 58.4
- 96.1
- 101
- 102
-53.7
- 67.9
- 57.9
- 96.5
- 99.6
- 112
- 70.1
- 72.0
- 61.8
* All values shown are %Bo [= (ODsample/ODnegatlve control)xlOO]
4015-10
Revision 0
January 1995
-------
TABLE 5
Bias of the EnviroGard,, 2,4-D in Soil Test Kit
Soi1# Fortification (ppm) Extraction^ Recovered (ppm)* % Recovery
SI
SI
SI
SI
SI
SI
Average >»
S2
S2
S2
S2
S2
S2
Average >»
S3
S3
S3
S3
S3
S3
Average >»
0.3
0.3
0.3
2
2
2
•»»»»»»»:
0.3
0.3
0.3
2
2
2
»»»»»»»:
0.3
0.3
0.3
2
2
2
»»»»»»»;
1
2
3
1
2
3
>»>»»»»»»:
1
2
3
1
2
3
>>»»»»»»»:
1
2
3
1
2
3
>»»»>»»»»x
0.21
0.24
0.23
1.87
2.12
2.40
»»»»»»»»:
0.29
0.29
0.30
2.05
1.89
2.22
>»»»»»>»»:
0.31
0.31
0.31
2.28
2.30
2.24
>»»>»»»»»:
70.0
80.0
76.6
93.5
106
120
»» 91.0
96.7
96.7
100
102
94.5
111
»» 100
103
103
103
114
115
112
»» 108
Overall Average %Recovery =99.7
4015-11
Revision 0
January 1995
-------
Table 6
Probability of False Negative and False Positive Results for 2,4-D RaPIDw
Assay kit at a 10 ppm Action Level in TCLP Extract from Organic Soil
Spike Concentration
2,4-D (PPM)
5
7.5
10
15
Probability of False
Positive (%)
0
70
N/A
N/A
Probability of False
Negative (%)
N/A
N/A
0
0
Results were based on ten replicate spiked samples. Cutoff
levels were established using 30 replicates of each solution
tested in 3 immunoassay batch runs.
N/A = No false positives possible above/below the
1imit.
action
4015-12
Revision 0
January 1995
-------
T
2,4-D Spiking Results on
ID #
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
Matrix/Spike
TCLP Buffer
TCLP Buffer + 15 ppm
TCLP Buffer + 10 ppm
TCLP Buffer + 5 ppm
Sandy Extract13
Sandy Extract + 15 ppm
Sandy Extract + 10 ppm
Sandy Extract + 5 ppm
Organic Extract0
Organic Extract + 15
Organic Extract + 10
Organic Extract + 5
Effluent #1
Effluent #1 + 15 ppm
Effluent #1 + 10 ppm
Effluent #1+5 ppm
Effluent n
Effluent #2 + 15 ppm
Effluent #2 + 10 ppm
Effluent #2+5 ppm
Runoff
Runoff + 15 ppm
Runoff + 10 ppm
Runoff + 5 ppm
able 7
\queous Environmental Matrices3
1
2,4-D Test Results
Rl
-
+
+
-
-
+
+
-
-
+
+
-
_
+
+
-
_
+
+
-
-
+
+
-
R2
_
+
+
-
-
+
+
-
-
+
+
-
_
+
+
-
-
+
+
-
-
+
+
-
R3
_
+
+
-
-
+
+
-
-
+
+
-
_
+
+
-
-
+
+
-
_
+
+
-
R4
-
+
+
-
-
+
+
-
-
+
+
-
_
+
+
-
-
+
+
-
-
+
+
-
R5
_
+
+
-
-
+
+
-
-
+
+
-
_
+
+
-
_
+
+
-
_
+
+
-
%POS
-
+
+
-
-
+
+
-
-
+
+
-
_
+
+
-
_
+
+
-
-
+
+
-
%NEG
-
+
+
-
-
+
+
-
-
+
+
-
_
+
+
-
_
+
+
-
_
+
+
-
4015-13
Revision 0
January 1995
-------
Table 7
2,4-D Spiking Results on Aqueous Environmental Matrices3
25
26
27
28
Municipal Water
Municipal Water + 140 ppb
Municipal Water + 70 ppb
Municipal Water + 35 ppb
2,4-D Test Results
_
+
+
-
_
+
+
-
_
+
+
-
_
_
_
-
_
-
_
-
_
_
_
-
_
-
_
-
i
8 2,4-D RaPIDw Assay kit
b Sandy Soil TCLP Extract
c Organic Soil TCLP Extract
4015-14
Revision 0
January 1995
-------
Table 8
2,4-D Spiking Results
2,4-D RaPIDM Assay kit vs. Method 8151
ID*
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
Matrix/Spike
TCLP Buffer
TCLP Buffer + 15 ppm
TCLP Buffer + 10 ppm
TCLP Buffer + 5 ppm
Sandy Extract3
Sandy Extract + 15 ppm
Sandy Extract + 10 ppm
Sandy Extract + 5 ppm
Organic Extract13
Organic Extract + 15 ppm
Organic Extract + 10 ppm
Organic Extract + 5 ppm
Effluent #1
Effluent #1 + 15 ppm
Effluent #1 + 10 ppm
Effluent #1+5 ppm
Effluent #2
Effluent #2 + 15 ppm
Effluent #2 + 10 ppm
LEf fluent #2+5 ppm
Runoff
Runoff + 15 ppm
Runoff + 10 ppm
Immunoassay
Results
5/5 Negative
5/5 Positive
5/5 Positive
5/5 Negative
5/5 Negative
5/5 Positive
5/5 Positive
5/5 Negative
5/5 Negative
5/5 Positive
5/5 Positive
5/5 Negative
5/5 Negative
5/5 Positive
5/5 Positive
5/5 Negative
5/5 Negative
5/5 Positive
5/5 Positive
5/5 Negative
5/5 Negative
5/5 Positive
5/5 Positive
Method 8151
2,4-D (ppm)
nd
13.0
11.0
5.6
nd
*
5.9, 5.2
*
nd
*
10.0, 9.5
*
*
*
11.0, 7.8
3.6
*
11.0
8.8, 9.5
*
nd
*
9.7, 8.6
Correlation
IA vs. GC
Yes
Yes
Yes
Yes
Yes
*
No
*
Yes
*
Yes
*
*
*
Yes
Yes
*
Yes
Yes
*
Yes
*
Yes
4015-15
Revision 0
January 1995
-------
Table 8
2,4-0 Spiking Results
2,4-D RaPID,, Assay kit vs. Method 8151
ID#
24
25
26
27
28
Matrix/Spike
Runoff + 5 ppm
Municipal Water
Municipal Water + 140 ppb
Municipal Water + 70 ppb
Municipal Water + 35
-
Immunoassay
Results
5/5 Negative
5/5 Negative
5/5 Positive
5/5 Positive
5/5 Negative
Method 8151
2,4-D (ppm)
5.5
nd
*
58.59 (ppb)
*
Correlation
IA vs. GC
Yes
N/A
N/A
N/A
N/A
Sandy Soil TCLP Extract
b Organic Soil TCLP Extract
nd non-detectable
N/A Not applicable to wastewater regulatory limit
* No analysis with Method 8150
4015-16
Revision 0
January 1995
-------
TABLE 9
Comparison of the EnviroGardm 2,4-D in Soil Test Kit to Method 8151
Interpretation of Results at 200 jug/kg
Sample #
Method 8150 ug/kg
Immunoassay Result
Agrees?
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
<200
<200
220
<200
<200
330
<200
<200
830
<200
310
350
<200
<200
200
<200
<200
440
560
380
NEGATIVE
NEGATIVE
POSITIVE
NEGATIVE
NEGATIVE
POSITIVE
POSITIVE
NEGATIVE
POSITIVE
NEGATIVE
POSITIVE
POSITIVE
NEGATIVE
NEGATIVE
POSITIVE
NEGATIVE
NEGATIVE
POSITIVE
POSITIVE
POSITIVE
YES
YES
YES
YES
YES
YES
FALSE POSITIVE
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
4015-17
Revision 0
January 1995
-------
21 <200 NEGATIVE YES
22 360 POSITIVE YES
23 <200 NEGATIVE YES
24 <200 NEGATIVE YES
25 <200 NEGATIVE YES
26 <200 NEGATIVE YES
27 <200 NEGATIVE YES
28 <200 NEGATIVE YES
29 <200 NEGATIVE YES
30 <200 NEGATIVE YES
4015-18 Revision 0
January 1995
-------
METHOD 4020
SCREENING FOR POLYCHLORINATED BIPHENYLS BY IMMUNOASSAY
1.0 SCOPE AND APPLICATION
1.1 Method 4020 is a procedure for screening soils and non-aqueous waste
liquids to determine when total polychlorinated biphenyls (PCBs) are present at
concentrations above 5, 10 or 50 mg/kg. Method 4020 provides an estimate for the
concentration of PCBs by comparison with a standard.
1.2 Using the test kit from which this method was developed, 95% of soil
samples containing 0.625 ppm or less of PCBs will produce a negative result in
the 5 ppm test configuration. Using another commercially available test kit, 97%
of soil samples containing 0.25 ppm or less of PCBs will produce a negative
result in the assay and greater than 99% of the samples containing 1.0 ppm or
more will produce a positive result. Tables 2-5, 7, 10, and 11 present false
positive and false negative data generated from commercially available test kits.
Using a test kit commercially available for screening non-aqueous waste liquids,
>95% of samples containing 0.2-0.5 ppm or less of PCB will produce a negative
result.
1.3 In cases where the exact concentrations of PCBs are required,
quantitative techniques (i.e., Method 8082) should be used.
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 Test kits are commercially available for this method. The
manufacturer's directions should be followed. In general, the method is
performed using a sample extract. Sample and an enzyme conjugate reagent are
added to immobilized antibody. The enzyme conjugate "competes" with PCB present
in the sample for binding to immobilized anti-PCB antibody. The test is
interpreted by comparing the response produced by testing a sample to the
response produced by testing standard(s) simultaneously.
3.0 INTERFERENCES
3.1 Chemically similar compounds and compounds which might be expected to
be found in conjunction with PCB contamination were tested to determine the
concentration required to produce a positive test result. These data are shown
in Tables 1A, IB, 1C, and ID.
4.0 APPARATUS AND MATERIALS
4.1 Immunoassay test kit: PCB RISc™ (EnSys, Inc.), EnviroGard™ PCB in
Soil (Millipore, Inc.), D TECH™ PCB test (Strategic Diagnostics Inc.), PCB
4020-1 Revision 0
January 1995
-------
RISc™ Liquid Waste Test System (EnSys, Inc.), or equivalent. Each commercially
available test kit will supply or specify the apparatus and materials necessary
for successful completion of the test.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1. Also refer to Reference 9 for the collection and handling of non-
aqueous waste liquids.
6.2 Samples may be contaminated, and should therefore be considered
hazardous and handled accordingly.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance specifications indicated
in Tables 2-11.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for the test kit being used for
quality control procedures specific to the test kit used. Additionally, guidance
provided in Method 4000 and Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.3 Do not use test kits past their expiration date.
8.4 Do not use tubes or reagents designated for use with other test kits.
8.5 Use the test kits within their specified storage temperature and
operating temperature limits.
8.6 Method 4020 is intended for field or laboratory use. The appropriate
level of quality assurance should accompany the application of this method to
document data quality.
9.0 METHOD PERFORMANCE
9.1 A study was conducted with the PCB RISc™ test kit using fourteen
4020-2 Revision 0
January 1995
-------
standard soils and three soil samples whose PCB concentration had been
established by Method 8082. Replicates were performed on seven of the standard
soils and on one of the soil samples for a total of 25 separate analyses. Each
of two different analysts ran the 25 analyses. Results indicated that "<"
assignments are accurate with almost 99% certainty at the 50 ppm level while ">"
assignments can be up to about 96% inaccurate as the sample concentration
approaches that of the testing level. Corresponding certainties at the 5 ppm
level are 92% and 82% respectively. Tables 2 and 3 summarize these results.
9.2 Table 4 presents method precision data generated using the PCB RISc™
test kit, comparing immunoassay test results with results obtained using Method
8082.
9.3 Method precision was determined with the EnviroGard PCB in Soil test
kit by assaying 4 different soils (previously determined to contain 5.04, 9.78,
11.8, and 25.1 mg/kg by Method 8082), at three different sites, using three
different lots of assay kits, three times a day for 9 days. A total of 81
analyses were performed for each soil. Error attributable to site, lot, date,
and operator were determined. Separately, the relative reactivity of Aroclors
1242, 1248, 1254, and 1260 were determined. Based on Aroclor heterogeneity, and
method imprecision, concentrations of Aroclor 1248 were selected that would
result in greater than 99% confidence for negative interpretation. A study was
conducted (Superfund SITE demonstration) on 114 field samples whose PCB
concentration were also determined by Method 8082. 32 of the field samples were
collected in duplicate (as coded field duplicates) and assayed by standard and
immunoassay methods. The results for all 146 samples are summarized in Tables
5 and 6.
9.4 Grab samples were obtained from sites in Pennsylvania, Iowa and
Illinois using a stainless steel trowel. Each sample was homogenized by placing
approximately six cubic inches in a stainless steel bucket and mixing with the
trowel for approximately two minutes. The soils was aliquotted into 2 six ounce
glass bottles. The samples were tested on site using the D TECH PCB test kit,
and sent to an analytical laboratory for analysis by Method 8082. These data are
compared in Table 7.
9.5 Tables 8 and 9 present data on the inter- and intra-assay precision
of the PCB RISc™ Liquid Waste Test System. The data were generated using 11
samples, each spiked at 0, 0.2 and 5 ppm, and assayed 4 times.
9.6 Tables 10 and 11 provide data from application of the PCB RISc™
Liquid Waste Test System to a series of liquid waste samples whose PCB
concentration had been established by Method 8082.
10.0 REFERENCES
1. J.P. Mapes, T.N. Stewart, K.D. McKenzie, L.R. McClelland, R.L. Mudd, W.B.
Manning, W.B. Studabaker, and S.B. Friedman, "PCB-RISc™ - An On-Site
Immunoassay for Detecting PCB in Soil", Bull. Environ. Contain. Toxicol.
(1993) 50:219-225.
2. PCB RISc™ Users Guide, Ensys Inc.
4020-3 Revision 0
January 1995
-------
3. R.W. Counts, R.R. Smith, J.H. Stewart, and R.A. Jenkins, "Evaluation of PCB
Rapid Immunoassay Screen Test System", Oak Ridge National Laboratory, Oak
Ridge, TN 37831, April 1992, unpublished
4. EnviroGard PCB in Soil Package Insert, Millipore Corp. 2/93.
5. Technical Evaluation Report on the Demonstration of PCB Field Screening
Technologies, SITE Program. EPA Contract Number 68-CO-0047. 2/93.
6. D TECH™ PCB Users Guide , SDI/Em Sciences
7. Melby, J.M., B.S. Finlin, A.B. McQuillin, H.G. Rovira, J.W. Stave, "PCB
Analysis by Enzyme Immunoassay", Strategic Diagnostics Incorporated,
Newark, Delaware, 1993
8. Melby, J.M., B.S. Finlin, A.B. McQuillin, H.G. Rovira, "Competitive
Enzyme Immunoassay (EIA) Field Screening System for the Detection of
PCB", 1993 PCB Seminar, EPRI, September 1993
9. T.A. Bellar and J.J Lichtenberg. The Analysis of Polychloringated
Biphenyls in Transformer Fluid and Waste Oils. U.S. EPA Research and
Development, EPA/EMSL-ORD, Cincinnati, Ohio (June 24, 1980). Revised
June 1981, EPA 600/4-81-045.
10. PCB RISc™ Liquid Waste Test System, User's Guide, EnSys Environmental
Products, Inc.
4020-4 Revision 0
January 1995
-------
TABLE 1A
CROSS REACTIVITY OF DIFFERENT COMPOUNDS"
Compound
1-Chloronaphthalene
1,2,4-Trichlorobenzene
2,4-Dichlorophenyl-benzenesulfonate
2,4-Dichloro-l-naphthol
Bifenox
Diesel fuel
Pentachlorobenzene
2,5-Dichloroanil ine
Hexachlorobenzene
Gasol ine
Dichlorofenthion
Tetradifon
Soil Equivalent Concentration (ppm)
Required to Yield a Positive Result
10,000
10,000
1,000
>10,000
500
>10,000
>10,000
>10,000
>10,000
>10,000
10,000
125
(a) PCB RISc'M test kit, Ensys, Inc. publication
4020-5
Revision 0
January 1995
-------
TABLE IB
CROSS REACTIVITY OF DIFFERENT COMPOUNDS'
Compound
Aroclor 1248
Aroclor 1242
Aroclor 1254
Aroclor 1260
1,2-, 1,3-, & 1,4-Dichlorobenzene
1,2,4-Trichlorobenzene
biphenyl
2,4-dichlorophenol
2,5-dichlorophenol
2,4,5-trichlorophenol
2,4,6-trichlorophenol
Pentachlorophenol
% Cross Reactivity
100
50
90
50
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
" EnviroGard PCB Test Kits (Millipore Corporation)
4020-6
Revision 0
January 1995
-------
TABLE 1C
CROSS REACTIVITY OF DIFFERENT COMPOUNDS"
Compound
Aroclor 1016
Aroclor 1221
Aroclor 1232
Aroclor 1242
Aroclor 1248
Aroclor 1254
Aroclor 1260
Aroclor 1262
Aroclor 1268
MDLb
(ppm)
5.7
25.5
9.0
1.5
0.8
0.5
0.75
0.5
3.8
1C 50C
(ppm)
83
300
105
31
24
10
10
10
40
% CROSS REACTIVITYd
12
3
10
32
42
100
100
100
25
METHOD: The compounds listed were assayed at various concentrations and
compared against an inhibition curve generated using Aroclor
1254. The concentration of the compound required to elicit a
positive response at the MDL as well as the concentration
required to yield 50% inhibition compared to the standard curve
were determined.
D TECH'M PCB test kit
The IC50 is defined as the concentration of compound required to produce a
test response equivalent to 50% of the maximum response.
c The Minimum Detection Limit (MDL) is defined as the lowest concentration of
compound that yields a positive test result.
d % Crossreactivity is determined by dividing the equivalent Aroclor 1254
concentration by the actual compound concentration at IC50
4020-7
Revision 0
January 1995
-------
TABLE ID
CROSS REACTIVITY OF DIFFERENT COMPOUNDS'
Compound
1- Chi oron aphtha! ene
1,2,4-Trichlorobenzene
2,4-Dichloro-l-naphthol
Bifenox
Pentachl orobenzene
2,5-Dichloroanil ine
Hexachlorobenzene
Dichlorofenthion
Tetradifon
% Cross-Reactivity
0.05%
0 . 05%
<0.20%
<0.10%
<0.05%
<0.05%
<0.05%
0.05%
<0.10%
Soil Equivalent
Concentration (ppm) Required
to Yield a Positive Result
10,000
10,000
>10,000
500
>10,000
>10,000
>10,000
10,000
125
(a) PCB RISc'M Liquid Waste Test System, Ensys, Inc.
4020-8
Revision 0
January 1995
-------
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Table 4
Comparison of PCB RISc Test Kit with GC
Sample 10
101
284
292
199
264
257
259
265
200
170
198
172
169
171
202
163
165
168
166
164
204
253
203
258
106
161
167
Screening Test
Results
<5 ppm
<5 ppm
<5 ppm
<5 ppm
<5 ppm
<5 ppm
<5 ppm
<5 ppm
<5 ppm
5-50
<5 ppm
5-50
5-50
5-50
<5 ppm, 5-50
5-50
5-50
5-50
5-50
5-50
5-50
5-50
5-50
5-50
5-50
5-50
5-50
GC Results
<0.5 ppm
<0.5 ppm
<0.5 ppm
0.5 ppm
1 ppm
1.8 ppm
4 ppm
4.5 ppm
5 ppm
5.8 ppm
2.2-5.8 ppm
6.2 ppm
7.2 ppm
7.2 ppm
1.3-7.2 ppm
8.7 ppm
9 ppm
9 ppm
9.3 ppm
11.9 ppm
12.8 ppm
13 ppm
13.5 ppm
15 ppm
15-19 ppm
15.3 ppm
16.2 ppm
AGREEMENT8
Y, FP, FN
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
4020-10
Revision 0
January 1995
-------
Table 4
Comparison of PCB RISc Test Kit with GC
Sample 10
247
148
205
162
175
176
197
243
252
178
201
254
238
248
250
242
256
249
245
241
246
261
240
267
239
104
108
Screening Test
Results
5-50
>50
5-50
5-50
5-50
5-50
5-50
5-50
5-50
5-50
5-50
5-50, >50
>50
5-50
>50
5-50
>50
>50
>50
5-50
>50
>50
>50
>50
>50
>50
>50
GC Results
18 ppm
18-34 ppm
20 ppm
20.4 ppm
21.2 ppm
21.6 ppm
32 ppm
32 ppm
32 ppm
43.7 ppm
43 ppm
56 ppm
46-60 ppm
44-60 ppm
68 ppm
30-69 ppm
73 ppm
96 ppm
102 ppm
154 ppm
154 ppm
204 ppm
251 ppm
339 ppm
460 ppm
200-3772 ppm
531-1450 ppm
AGREEMENT"
Y, FP, FN
Y
FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
FN
Y
Y
Y
Y
Y
Y
Y
Y=Yes, FN=False Negative, FP=False Positive
4020-11
Revision 0
January 1995
-------
Table 5
Comparison of EnviroGard PCB Kit with GC
SAMPLE
NUMBER
001
002
003
004
005
006
007
008
009
010
Oil
012
013
014
015
015D
016
017
018
019
020
021
022
022D
023
024
SCREENING GC RESULT0
RESULTc'd [8082]
>10 5.98
>10 1.27
<10 0.11
>10 6.71
>10 1.37
>10 0.68
>10 0.55
>10 2.00
>10 1.30
>10 0.17
>10 1.15
<10 NDf
<10 1.13
<10 0.18
>10 9.13
>10 9.84
>10 2110
>10 2.55
>10 45.4
>10 6.70
<10 0.07
<10 0.06
<10 0.54
<10 0.72
>10 20.8
<10 0.06
AGREEMENT
Y, FN, FP
ppg
FP
Y
ppg
FP
FP
FP
FP
FP
FP
FP
Y
Y
Y
FPa
ppg
Y
FP
Y
ppg
Y
Y
Y
Y
Y
Y
4020-12
Revision 0
January 1995
-------
Table 5 (continued)
SAMPLE
NUMBER
024D
025
026
027
028
028D
029
030
031
032
033
034
035
035D
036
037
037D
038
039
040
041
042
042D
043
043D
044
SCREENING GC RESULT0
RESULT" [8082]
<10 0.05
>10 11.7
<10 1.96
<10 0.06
<10 0.22
<10 0.22
<10 0.23
<10 1.15
<10 0.26
>10 47.6
>10 6.00
>10 34.0
<10 NDf
<10 NDf
>10 816
<10 0.06
<10 0.04
>10 1030
<10 0.68
>10 4.25
<10 ND'
>10 0.52
>10 0.47
>10 1.69
>10 1.74
<10 0.59
AGREEMENT
Y, FN, FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
FP9
Y
Y
Y
Y
Y
Y
Y
Y
FP
Y
FP
FP
FP
FP
Y
4020-13
Revision 0
January 1995
-------
Table 5 (continued)
SAMPLE
NUMBER
045
046
046D
047
047D
048
049
050
050D
051
052
053
054
055
056
057
058
059
060
060D
061
062
063
063D
064
065
SCREENING GC RESULT0
RESULTc'd [8082]
<10 NDf
<10 NDf
<10 NDf
<10 0.09
<10 0.10
<10 NDd
<10 NDd
>10 3.60
>10 4.41
<10 NDf
>10 4.21
<10 0.96
<10 0.52
<10 2.40
<10 0.51
<10 NDf
<10 0.69
>10 7.86
>10 0.62
<10 0.58
>10 580
>10 2.35
<10 0.09
<10 0.15
>10 19.0
>10 3.08
AGREEMENT
Y, FN, FP
Y
Y
Y
Y
Y
Y
Y
FP
FP
Y
FP
Y
Y
Y
Y
Y
Y
FP°
FP
Y
Y
FP
Y
Y
Y
FP
4020-14
Revision 0
January 1995
-------
Table 5 (continued)
SAMPLE
NUMBER
066
067
068
069
069D
070
071
071D
072
073
074
075
076
077
078
079
080
081
081D
082
082D
083
083D
084
084D
085
SCREENING GC RESULT0
RESULT°'d [8082]
<10 1.98
<10 0.08
<10 0.50
<10 NDf
<10 NDf
<10 NDf
<10 0.05
<10 NDf
<10 0.04
>10 15.8
>10 13.3
>10 23.0
>10 46.7
<10 NDf
>10 2.27
>10 42.8
<10 3.77
<10 0.69
<10 0.45
<10 NDf
<10 0.24
<10 0.48
<10 0.41
>10 1.16
>10 1.08
>10 428
AGREEMENT
Y, FN, FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
FP
Y
Y
Y
Y
Y
Y
Y
Y
FP
FP
Y
4020-15
Revision 0
January 1995
-------
Table 5 (continued;
SAMPLE
NUMBER
085D
086
086D
087
087D
088
088D
089
090
090D
091
091D
092
092D
093
094
095
095D
096
097
097D
098
098D
099
100
100D
SCREENING GC RESULT0
RESULTc'd [8082]
>10 465
<10 1.42
<10 1.25
<10 0.08
<10 NDf
>10 2.70
>10 1.77
>10 45.0
<10 1.01
<10 1.40
>10 1630
>10 1704
<10 1.21
<10 NDf
<10 0.30
<10 0.36
>10 17.5
>10 31.2
<10 0.06
<10 1.23
<10 0.29
>10 1.17
>10 0.83
<10 NDf
>10 177
>10 167
AGREEMENT
Y, FN, FP
Y
Y
Y
Y
Y
FP
FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
FP
FP
Y
Y
Y
4020-16
Revision 0
January 1995
-------
Table 5 (continued)
SAMPLE
NUMBER
101
102
102D
103
104
105
106
107
108
109
109D
110
111
112
113
114
SCREENING GC RESULT0
RESULT0'" [8082]
>10 1.21
>10 293
>10 177
>10 40.3
>10 7.66
<10 0.21
<10 2.50
>10 14.1
>10 3.84
<10 NDf
<10 NDf
<10 NDf
<10 NDf
>10 315
>10 14.9
>10 66.3
AGREEMENT6
Y, FN, FP
FP
Y
Y
Y
ppa
Y
Y
Y
FP
Y
Y
Y
Y
Y
Y
Y
c mg/kg (ppm)
d Screening Calibrator is 5 mg/kg Aroclor 1248
e Y=Yes, FN=False Negative, FP=False Positive
f ND = Not Detectable
8 Expected Result Based on Calibrator Concentration
4020-17
Revision 0
January 1995
-------
Table 6
EnviroGard PCB Kit Field Performance Summary
Specificity: [l-(Reported Positives/True Negatives)] = [l-(37/109)] = 66%
Note 1: 8 of the 37 reported positive samples had PCB contamination
levels between 5 and 10 mg/kg. Soils in this range should test
"positive" because the assay calibrator is 5 mg/kg Aroclor 1248.
A positive assay bias is necessary to prevent false negative
results.
Eliminating these samples from the calculations produces a
Specificity of:
[l-(Reported Positives/True Negatives)] = [l-(29/101)] = 71%
Note 2: The distribution of false positives is not random (p < 0.05),
with a clustering at the beginning of the sample set. This
observation was included in Developers Comments which were added
to the final draft of the Technical Evaluation Report2. One
explanation for the higher frequency of false positive results
at the beginning is inexperience of the operator with the
method. If the first 20 samples are eliminated from the
Specificity analysis, the following result is obtained:
[l-(Reported Positives/True Negatives)] = [l-(20/86)] = 77%
In the SITE demonstration, the PCB Immunoassay had a 77%
positive predictive value.
Sensitivity: [l-(Reported Negatives/True Positives)] = [1-(0/31)] = 100%
In the SITE demonstration, the PCB Immunoassay had a 100%
negative predictive value.
4020-18 Revision 0
January 1995
-------
TABLE 7
COMPARISON OF D TECH PCB test kit WITH GC - TRIAL #1
SAMPLE
01
02
J3
J5
J6
07
J8
J9
J10
Jll
J12
J13
J14
J15
016
J17
J18
J19
J20
J21
J22
J23
J24
D TECH
(ppm)
4.0-15
>50
15-50
15-50
>50
4.0-15
4.0-15
>50
>50
>50
15-50
>50
>50
15-50
15-50
>50
>50
>50
>50
>50
1.0 .
1.0
<0.5
GC
(ppm)
5.0
147
54
160
1200
12
28
463
1760
28
17
1300
186
31
36
31
130
1310
2620
11100
0.01
0.60
0.10
AGREEMENT
Y, FN, FP
Y
Y
Y
FN
Y
Y
FN
Y
Y
FP
Y
Y
Y
Y
Y
FP
Y
Y
Y
Y
FP
Y
Y
SAMPLE
J25
J26
J28
J28
J29
J30
J31
J32
J33
J34
035
036
037
038
039
040
041
042
043
044
045
046
047
D TECH
(ppm)
0.5
<0.5
1.0
<0.5
0.5
>50
4.0-15
0.5
0.5
1.0
1.0
>50
<0.5
0.5
0.5
<0.5
<0.5
1.0
1.0
15-50
15-50
<0.5
<0.5
GC
(ppm)
0.12
0.01
1.8
0.18
0.54
21
13
0.72
0.32
0.36
0.26
70
0.12
0.81
0.33
0.19
0.01
0.43
0.31
503.4
5.6
0.02
0.22
AGREEMENT
Y, FN, FP
FP
Y
Y
Y
Y
FP
Y
Y
Y
FP
FP
Y
Y
Y
Y
Y
Y
FP
FP
FN
FP
Y
Y
Y=Yes, FN=False Negative, FP=False Positive
4020-19
Revision 0
Oanuary 1995
-------
TABLE 7(cont)
COMPARISON OF D TECH PCB test kit WITH GC - Trial #2
SAMPLE
Gl
G2
G3
G4
G5
G6
G7
G8
G9
G10
Gil
G12
G13
G14
G15
G16
G17
G18
G19
G20
D TECH
(ppm)
15-50
4.0-15
1.0-4.0
15-50
<0.5
1.0-4.0
1.0-4.0
15-50
4.0-15
15-50
4.0-15
4.0-15
4.0-15
0.5-1.0
<0.5
1.0-4.0
4.0-15
4.0-15
1.0-4.0
>50
GC
(ppm)
18
11
3.4
6.5
0.01
1.4
0.30
7.5
33
8
11
24
4.3
1.3
0.01
3.2
18
4.6
2.3
37
AGREEMENT
Y, FN, FP
Y
Y
Y
FP
Y
Y
FP
FP
FN
FP
Y
FN
Y
Y
Y
Y
Y
Y
Y
FP
4020-20
Revision 0
January 1995
-------
TABLE 7(cont)
COMPARISON OF D TECH PCB test kit WITH GC - Trial #3
SAMPLE
W1A
W2A
W3A
W4A
W5A
W6A
W7A
W8A
W9A
W10A
W11A
W12A
W13A
W14A
W15A
W16A
W17A
W18A
W19A
W20A
W21A
W22A
W23A
D TECH
(ppm)
4.0-15
4.0-15
1.0-4.0
4.0-15
>50
>50
>50
4.0-15
1.0-4.0
0.5-1.0
15-50
15-50
15-50
4.0-15
1.0-4.0
1.0-4.0
4.0-15
1.0-4.0
4.0-15
>50
>50
1.0-4.0
>50
GC
(ppm)
9.1
11
2.8
13
29
1200
57
18
1.3
0.44
120
48
19
2.7
1.3
0.3
1.4
2.2
8.2
9.3
110
0.6
46
AGREEMENT
Y, FN, FP
Y
Y
Y
Y
FP
Y
Y
Y
Y
Y
FN
Y
Y
Y
Y
FP
FP
Y
Y
FP
Y
Y
Y
4020-21
Revision 0
January 1995
-------
Table 8
Intraassay Precision of the PCB RISc™ Liquid Waste Test System
PCB 1248 Spike
Concentration
(ppm)
0
0.2
5
Signal %RSD
(OD450nm) N=44
(11 data sets)
6.4%
5.9%
7.9%
Statistical Percentage of
False Results Compared to
Standards
<0.02%
4.1%
1.4%
Table 9
Interassay Precision of the PCB RISc™ Liquid
Waste Test System
PCB 1248 Spike
Concentration (ppm)
0
0.2
5
Signal %RSD
(OD450nm) N=44
(11 data sets)
6.4%
8.3%
8.5%
4020-22
Revision 0
January 1995
-------
Table 10
Comparison of PCB RISc™ Liquid Waste Test with Method 8082
Sample
ID
302
303
304
306
307
308
310
311
331
380
381
382
383
384
385
387
388
389
390
391
394
395
396
398
399
400
401
402
403
404
Sample Matrix
Condensate
Condensate
Condensate
Condensate
Condensate
Condensate
Condensate
Condensate
Transformer Oil
Transformer Oil
Transformer Oil
Transformer Oil
Transformer Oil
Transformer Oil
Transformer Oil
Coolant
2,4-D Rinse Water
Waste Solvent
Herbicide
Paint/Solvent
Waste Solvent
Waste Solvent
Waste Oil
Chlor. Solvent
Paint
Pump Oil
Waste Solvent
Herbicide
Paint/Solvent
Printing Solvent
GC Results
Aroclor
NDb
ND
1242
1242
1242
1242
1254
1242
1260
PCBC
PCB
PCB
PCB
PCB
PCB
PCB
1254
1242
ND
1254
1242/1260
1242/1260
1260
ND
ND
ND
ND
ND
ND
ND
Cone, ppm
ND
ND
25
5
<10
58
25
200
183
20
38
163
176
336
6400
10
<10
29
<2
9
11/17
2/2
323
<5
<50
<50
<35
<50
<5
<5
IA Results
Test
Results
<5
<5
>5
>5
<5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
<5
>5
<5
>5
>5
<5
>5
<5
<5
<5
<5
<5
<5
<5
Corr.
with GC
Results
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
4020-23
Revision 0
January 1995
-------
Table 10 (continued)
Sample
ID
405
407
408
409
410
411
412
413
414
415
416
417
418
419
420
421
422
423
424
425
Sample Matrix
Waste Solvent
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
Waste Oil
GC Results
Aroclor
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
PCB
ND
ND
ND
ND
ND
ND
ND
ND
ND
Cone, ppm
<50
ND
ND
ND
ND
ND
ND
ND
ND
ND
50
ND
ND
ND
ND
ND
ND
ND
ND
ND
Number of False Positive Results
Rate
Number of False Negative Results
Rate
IA Results
Test
Results
<5
>5
<5
<5
<5
<5
<5
<5
<5
<5
>5
<5
<5
<5
<5
<5
<5
<5
<5
<5
Corr.
with GC
Results
yes
Fpd
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
1/32
3 . 1%
0/18
0 . 0%
a Trial 1 data
b ND = Not Detectable
c PCB = Aroclor was not determined
d
FP = False positive
4020-24
Revision 0
January 1995
-------
Table 11
Correlation of PCB RISc™ Liquid Waste Test and Method 8082 Results
Using Spiked and Unspiked Liquid Waste Field Samples
ID
001
002
003
004
005
006
007
008
009
010
Oil
012
013
014
015
016
017
018
019
020
021
022
023
024
025
026
027
Matrix
Aromatic solvent
Aviation gas
Chiller oil
Compressor oil
Coolant + water
Coolant oil
Coolant oil
Cutting oil
Cutting oil
Degreaser still
bottom
Dope oil
Draw Lube oil
Fleet crankcase
oil
Floor sealer
Fuel oil
Hi-BTU oil
Honing oil
Hydraulic oil
Hydraulic oil
Hydraulic oil
Machine oil
Mineral oil
Mineral spirits
Mineral spirits +
ink
Mixed flammables
Mixed solvents
Naphtha
GC
Results
Unspiked
ppm
<5
<5
<5
<5
<5
NRb
NR
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
NR
<5
<5
<5
<5
<5
<5
Immunoassay Result
Unspiked
ppm
<5
<5
<5
<5
<5
NR
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
>5
<5
<5
<5
Spiked (5
ppm 1248)
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
NR
>5
>5
>5
>5
>5
>5
Interp.
FP
4020-25
Revision 0
January 1995
-------
Table 11 (continued)
ID
028
029
030
031
032
033
034
035
036
037
038
039
040
041
042
043
044
045
046
047
048
049
050
051
052
053
054
055
Matrix
Oil
Oil
Oil
Oil
Oil
Oil
Oil + 1,1,1-
trichloroethane
Oil sludge
Oil + freon
Oil + mineral
spirits
Oil + scum
solution
Oily water
Paint thinner
Paint thinner
Paint thinner
Paint waste
Paint waste +
thinner
Perce + oil
Petroleum
distillates
Petroleum naphtha
Pumping oil
RAC-1 SKOS
Sk oil
Sk oil
Smog Hog
Toluene + hexane
Toluene + stain
1,1,1-
Trichloroethane
GC
Results
Unspiked
ppm
n <5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
NR
<5
<5
<5
<5
<5
Immunoassay Result
Unspiked
ppm
<5
<5
<5
<5
<5
<5
<5
>5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
>5
<5
<5
<5
<5
<5
<5
<5
<5
>5
Spiked (5
ppm 1248)
^ >5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
Interp.
FP
FP
FP
4020-26
Revision 0
January 1995
-------
Table 11 (continued)
ID
056
057
058
059
060
061
062
063
064
065
066
067
068
069
070
071
072
073
074
075
076
077
078
079
080
081
082
Matrix
1,1,1-
Trichloroethane
1,1,1-
Trichloroethane
1,1,1-
Trichloroethane
1,1,1-TCE +
methanol
Trichloroethylene
Trichloroethylene
Trichloroethylene
Turpentine
Used n-
butyl acetate
Used oil + freon
Used oil + freon
Used oils
Used petroleum
Used petroleum
Used synthetic oil
Varnish + stain
Varsol
Waste coolant +
oil
Waste ink +
solvent
Waste naphtha
Waste oil
Waste oil
Waste oil
Waste oil
Waste oil
Waste oil
Waste oil
GC
Results
Unspiked
ppm
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
Immunoassay Result
Unspiked
ppm
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
Spiked (5
ppm 1248)
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
Interp.
4020-27
Revision 0
January 1995
-------
Table 11 (continued)
ID
083
084
085
086
087
088
089
090
091
092
093
094
095
096
097
098
099
100
Matrix
Waste oil
Waste oil
Waste oil +
kerosene
Waste oil + gas
Waste paint
Waste paint
Waste paint
Waste paint
Waste paint
Waste paint
Waste SC-49
solvent
Waste solvent
Waste stoddard
Waste toner
Waste tramp oil
Waste transmission
fluid
Xylene
Not Recorded
No. of False Positive
Results
Rate
No. of False Negative
Results
Rate
GC
Results
Unspiked
ppm
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
Immunoassay Result
Unspiked
ppm
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
>5
<5
6/99
6.1%
Spiked (5
ppm 1248)
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
>5
NR
Interp.
FP
FP
0.98
0 . 0%
a Trial 2 data
b NR = not run
4020-28
Revision 0
January 1995
-------
METHOD 4030
SOIL SCREENING FOR PETROLEUM HYDROCARBONS BY IMMUNOASSAY
1.0 SCOPE AND APPLICATION
1.1 Method 4030 is a procedure for screening soils to determine whether
total petroleum hydrocarbons (TPH) are likely to be present. Depending on the
testing product selected, samples may be used to locate samples with low (<40-100
ppm), medium, and high (>1000 ppm) concentrations of contaminates, or to
determine if TPH is present at concentrations above 5, 25, 100, or 500 mg/kg.
Method 4030 provides an estimate for the concentration of TPH by comparison
against standards, and can be used to produce multiple results within an hour of
sampling.
1.2 Using the test kit from which this method was developed, 95 % of
samples containing 25 ppm or less of TPH will produce a negative result in the
100 ppm test configuration.
1.3 The sensitivity of any immunoassay test depends on the binding of the
target analyte to the antibodies used in the kit. The testing product used to
develop this method is most sensitive to the small aromatic compounds (e.g.,
ethylbenzene, xylenes, and naphthalene) found in fuels. Refer to the package
insert of the testing product selected for specific information about
sensitivity.
1.4 The sensitivity of the test is influenced by the nature of the
hydrocarbon contamination and any degradation processes operating at a site.
Although the action level of the test may vary from site to site, the test should
produce internally consistent results at a particular site.
1.5 In cases where a more exact measurement of TPH concentration is
required, additional techniques (i.e., gas chromatography Method 8015 or infra-
red spectroscopy Method 8440) should be used.
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.
2.0 SUMMARY OF METHOD
2.1 Test kits are commercially available for this method. The
manufacturer's directions should be followed.
2.2 In general, the method is performed using an extract of a soil
sample. Filtered extracts may be stored cold, in the dark. An aliquot of the
extract and an enzyme-TPH conjugate reagent are added to immobilized TPH
antibody. The enzyme-TPH conjugate "competes" with hydrocarbons present in the
4030-1 Revision 0
January 1995
-------
sample for binding to immobilized anti-TPH antibody. The test is interpreted by
comparing the response produced by a sample to the response produced by a
reference reaction.
3.0 INTERFERENCES
3.1 Compounds that are chemically similar to petroleum hydrocarbons may
cause a positive test (false positive) for TPH. The data for the lower limit of
detection of these compounds are provided in Tables 1A and IB. Consult the
information provided by the manufacturer of the kit used for additional
information regarding cross reactivity with other compounds.
3.2 Storage and use temperatures may modify the method performance.
Follow the manufacturer's directions for storage and use.
3.3 Appropriate standards must be used (/.e., diesel standards for diesel
analysis, JP-4 for analysis of JP-4, etc.), or excessive false negative or false
positive rates may result.
4.0 APPARATUS AND MATERIALS
4.1 Immunoassay test kit: PETRO RISc Soil Test (EnSys, Inc.), EnviroGard™
Petroleum Fuels in Soil, (Millipore, Inc.), or equivalent. Each commercially
available test kit will supply or specify the apparatus and materials necessary
for successful completion of the test.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Soil samples may be contaminated, and should therefore be considered
hazardous and handled accordingly.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance specifications indicated
in Tables 2-12.
4030-2 Revision 0
January 1995
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7.2 Appropriate standards must be used to prevent excessive rates of
false negative or false positive results.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for the test kit being used
for quality control procedures specific to the test kit used. Additionally,
guidance provided in Method 4000 and Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.3 Do not use test kits past their expiration date.
8.4 Do not use tubes or reagents designated for use with other test kits.
8.5 Use the test kits within their specified storage temperature and
operating temperature limits.
8.6 Method 4030 is intended for field or laboratory use. The appropriate
level of quality assurance should accompany the application of this method to
document data quality.
9.0 METHOD PERFORMANCE
9.1 A single laboratory study was conducted with the PETRO RISc Soil Test,
EnSys, Inc., using five contaminated soil samples. The samples were contaminated
with oxygenated gasoline, oxygenated gasoline 24 hours after contamination, low
aromatic diesel (purchased in California), normal diesel (purchased in Northern
Virginia), and JP-4 jet fuel. Five replicate determinations were made using the
kits, and the data compared with values obtained using GC/FID (Method 8015) and
IR (Method 8440). Several different analysts ran the immunoassay analyses.
Samples two- to five-fold below the action level generally gave readings less
than the action level. Samples two fold above the action level gave readings
greater than the action level. Samples at or near the action level give mixed
results (e.g., both less than and greater than the action level). Tables 2 - 6
summarize these results.
9.2 Sensitivity of the EnviroGard Petroleum Fuels in Soil Test Kit was
determined by establishing the "noise" level expected from matrix effects
encountered in negative soil samples and determining the corresponding TPH
concentration by comparison to the analyte-specific response curve. 8 different
soils which did not contain TPH were assayed. Each of these soils was extracted
in triplicate and each extract was assayed in three different assays. The mean
and the standard deviation of the resulting %Bo's (%Bo = [(ODsample/ODnegatlve
control)xlOO]) were calculated and the sensitivity was estimated at two standard
deviations below the mean. The sensitivity for Method 4030 was determined to be
4030-3 Revision 0
January 1995
-------
80% Bo at a 95% confidence interval. Based on the average assay response to home
heating oil (HHO), this corresponds to 5.8 ppm. These data are shown in Table
7.
9.3 The effect of water content of the soil samples was determined by
assaying three different soil samples which had been dried and subsequently had
water added to 30% (w/w). Aliquots of these samples were then fortified with
HHO. Each soil sample was assayed three times, with and without added water, and
with and without HHO fortification. It was determined that water in soil up to
30% had no detectable effect on the method. These data are shown in Table 8.
9.4 The effect of the pH of the soil extract was determined by adjusting
the soil pH of three soil samples. Soil samples were adjusted to pH 2 - 4 using
6N HC1 and pH 10 - 12 using 6N NaOH. Aliquots of the pH adjusted soil samples
were fortified with home heating oil. Each soil sample was assayed unadjusted
and with pH adjusted to 2-4 and 10-12, both unfortified and fortified. These
extracts were assayed three times. It was determined that soil samples with pH
ranging from 2 to 12 had no detectable effect on the performance of the method.
These data are shown in Table 9.
9.5 Two field studies were conducted at contaminated sites using the PETRO
RISc Soil Test, EnSys, Inc.. In Field Trial 1, the method was used to locate
soil contamination resulting from a leaking above ground gasoline tank. In Field
Trial 2, the method was used to evaluate diesel fuel contamination in a railroad
contaminated soil, sludge, and wastewater impound. Overall, a high degree of
correlation was observed between the standard method and the immunoassay method.
The application of the immunoassay method to 23 samples (46 analyses) resulted
in eight conclusive false positive results (17%) and three conclusive false
negative results (7%). Tables 10 and 11 summarize these results. There was
agreement for 76% of the samples tested in the two trials for which data are
presented.
9.6 Two field trials were undertaken to investigate the ability of the
EnviroGard Petroleum Fuels in Soil Test Kit to identify soil samples which were
contaminated with TPH. In trial 1 the method was used to identify soil which was
contaminated with gasoline from leaking underground storage tanks. The
immunoassay was compared to Method 8015. Twenty samples were analyzed by both
methods. Interpreting the results at a cutoff of 100 ppm resulted in 1/20 (5%)
false negatives and 0/20 (0%) false positives. In trial 2, the method was used
to identify soil which was contaminated with JP-4 jet fuel from leaking
semi-submerged storage tanks. The immunoassay was compared to Method 8015. Ten
samples were analyzed by both methods. Interpreting the results at 1,000 ppm
resulted in 0/10 (0%) false negatives and 1/10 (10%) false positives. Overall,
for both field trials, there were 1/30 (3.3%) false negatives and 1/30 (3.3%)
false positives. These data are summarized in Table 12.
10.0 REFERENCES
1. PETRO RISc™ Users Guide, Ensys Inc.
4030-4 Revision 0
January 1995
-------
2. Marsden, P.J., S-F Tsang, and N. Chau, "Evaluation of the PETRO RISc™ kit
Immunoassay Screen Test System", Science Applications International
Corporation under contract to EnSys Inc., June 1992, unpublished
3. EnviroGard™ Petroleum Fuels in Soil Test Kit Guide, Millipore, Inc.
4030-5 Revision 0
January 1995
-------
Table 1A
CROSS REACTIVITY3
Compound
Gasol ine
Diesel fuel, regular #2
Jet A fuel
Kerosene
Fuel oil n
Mineral Spirits
Light lubricating oil
Lithium grease
Brake fluid
Chain lubricant
Toluene
o-Xylene
m-Xylene
p-Xylene
Ethyl benzene
Hexachlorobenzene
Trichloroethylene
Acenaphthene
Naphthalene
Creosote
2-Methylpentane
Hexanes, mixed
Heptane
iso-Octane
Undecane
Soil Equivalent Concentration (ppm)
Required to Yield a Positive Result
100
75
75
100
100
<30
7,000
10,000
>10,000
>10,000
200
50
100
300
50
<30
1,000
<30
<30
<30
150
250
300
30
>10,000
PETRO RISc Soil Test, EnSys, Inc.
4030-6
Revision 0
January 1995
-------
TABLE IB
CROSS REACTIVITY3
Compound
1,2,4 - Trimethyl benzene
m - Xylene
Acenaphthylene
Acenapthene
p - Xylene
Naphthalene
1,3,5 - Trimethyl benzene
Fl uorene
Phenanthrene
o - Xylene
Ethyl benzene
Toluene
Propyl benzene
Chlordane
Benzene
Toxaphene
Concentration Required
Positive Interpretation
for
(ppm)
0.1
0.3
0.3
0.4
0.5
0.7
2
2
2
3
5
7
11
45
70
70
The following compounds were tested and found to yield negative results
for concentrations up to 1000 ppm:
PCB (Aroclor 1248) TNT
Pentachlorophenol DDT
EnviroGard™ Petroleum Fuels in Soil, Millipore, Inc.
4030-7
Revision 0
January 1995
-------
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TABLE 7
METHOD SENSITIVITY
Part 1 - Average Response with Negative Soils
Soil# Soil Type Average %Bo (n = 9)Standard Deviation
SAND
S2
S3
S4
S5
S6
S7
S8
91.4
LOAM
CLAY
LOAM
CLAY
LOAM/SAND
SAND/LOAM
LOAM
4.1
83.1
84.4
80.9
89.7
91.2
89.0
90.0
3.2
3.1
1.3
1.7
0.2
0.3
1.4
AVERAGE 87.5 4.0
Part 2 - Average Response with Calibrators
Calibrator
Concentration (ppm) Average Absorbance Average %Bo
0
5
15
50
125
1.339
1.097
0.825
0.427
0.219
N/A
81.9
61.7
31.9
16.3
Part 3 - Method Sensitivity
Based on Part 1 and Part 2 Above:
Average %Bo - 2 SO = 79.6 which is equivalent to 5.8 ppm
Average %Bo - 3 SD = 75.6 which is equivalent to 7.0 ppm
(%Bo = [(ODsample/ODnegatlvecontro,)xlOO])
4030-13 Revision 0
January 1995
-------
TABLE 8
EFFECT OF WATER CONTENT IN SOIL SAMPLES
Soil % Water Fortified? Rep. 1* Rep. 2 Rep. 3 Mean Std. Dev. ± 2 SD Range
SI
SI
SI
SI
S2
S2
S2
S2
S3
S3
S3
S3
0
30
0
30
0
30
0
30
0
30
0
30
No
No
Yes
Yes
No
No
Yes
Yes
No
No
Yes
Yes
101
100
59
60
57
74
40
44
70
81
41
61
.3
.5
.2
.9
.5
.3
.5
.1
.5
.1
.3
99
115
65
74
53
91
40
67
85
109
46
76
.1
.5
.8
.7
.9
.8
.9
.8
.6
.4
.6
.7
111.8
109.1
69.6
83.1
72.3
85.2
45.6
68.4
76.7
103.4
60.7
63.1
104.1
108.4
64.9
72.3
61.4
83.8
42.3
60.2
77.5
98.1
49.5
67.0
6.8
7.5
5.3
11.7
9.7
8.7
2.9
13.6
7.8
14.7
10.1
8.4
90.4
93.4
49.9
49.2
42.0
66.4
36.5
33.0
61.9
68.7
29.3
50.2
- 117.7
- 123.4
- 75.5
- 96.0
- 80.8
- 101.2
-48.1
- 87.4
- 93.1
- 127.5
- 69.7
-83.8
* All values shown are %Bo [= (ODsample/ODn
egative coi
,n«roi)xlOO]
4030-14
Revision 0
January 1995
-------
TABLE 9
EFFECT OF pH ON SOIL SAMPLES
oil
SI
SI
SI
SI
SI
SI
S2
S2
S2
S2
S2
S2
S3
S3
S3
S3
S3
S3
oH Ad.i. Fortified? Rep.
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
No
No
No
Yes
Yes
Yes
No
No
No
Yes
Yes
Yes
No
No
No
Yes
Yes
Yes
88.
108.
101.
64.
52.
69.
76.
101.
89.
59.
68.
47.
83.
89.
80.
60.
58.
53.
1*
9
9
2
3
9
3
2
2
9
4
1
8
4
3
6
2
8
4
Rep. 2
93.2
66.0
90.3
55.7
41.1
61.7
86.4
82.4
72.1
60.3
62.3
51.7
88.4
84.9
84.2
53.6
58.5
41.8
Rep. 3
92.8
88.1
90.6
58.0
49.4
57.5
83.1
99.5
77.7
53.7
59.3
39.4
85.3
91.0
90.3
58.8
62.0
59.9
Mean Std. Dev.
91.6
87.7
94.0
59.3
47.8
62.8
81.9
94.4
79.9
57.8
63.2
46.3
85.7
88.4
85.0
57.5
59.8
51.7
2.4
21.5
6.2
4.5
6.1
6.0
5.2
10.4
9.1
3.6
4.5
6.3
2.5
3.1
4.9
3.5
1.9
9.2
± 2 SD
86
44
81
50
35
50
71
73
61
50
54
33
80
82
75
47
56
33
.8 -
.7-
.6 -
.3 -
.6 -
.8 -
.5 -
.6 -
.7 -
.6 -
.2 -
.7 -
.7 -
.2 -
.2 -
.7 -
.0 -
.3 -
Range
96.4
109.2
106.4
68.3
60.0
74.8
92.3
115.2
98.1
65.0
72.2
58.9
90.7
94.6
94.8
64.5
63.6
70.1
* All values shown are %Bo [= (ODsample/ODn
egative co
n,rol)XlOO]
4030-15
Revision 0
January 1995
-------
Table 10
PETRO RISc Soil Test
Field Trial 1
Sample ID
AST-01
AST-02
AST-03
AST-04
AST-05
AST-06
AST-07
AST-08
AST-09
IR Method (ppm)
<20
520
1700
130
20
40
400
640
1600
100 ppm Test
Result
< 100
> 100
> 100
> 100
> 100
> 100
> 100
> 100
> 100
AGREEMENT3
Y, FP, FN
Y
Y
Y
Y
FP
FP
Y
Y
Y
1000 ppm Test
Result
< 1000
> 1000
> 1000
< 1000
< 1000
< 1000
< 1000
< 1000
> 1000
AGREEMENT3
Y, FP, FN
Y
FP
Y
Y
Y
FN
FN
FN
Y
Y Immunoassay and GC or IR results agree
FP False Positive
FN False Negative
4030-16
Revision 0
January 1995
-------
Table 11
PETRO RISc Soil Test
Field Trial 2
Sample
ID
1-B
2-A
2-B
2-C
3-B
3-C
4-A
4-B
5-A
5-B
5-C
6-B
8
9
GC
Extractables
(ppm)
5720
610
370
2270
4870
760
66
303
20400
26300
267
550
59300
26500
TRPH
(ppm)
20800
14700
6800
1950
18600
1180
4100
2100
29600
28600
330
22700
64400
12900
75 ppm Test
Result
> 75
> 75
> 75
> 75
> 75
> 75
> 75
> 75
> 75
> 75
> 75
> 75
> 75
> 75
AGREEMENT3
Y, FP, FN
TRPH
Y
Y
Y
Y
Y
Y
FP"
Y
Y
Y
Y
Y
Y
Y
GC
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
750 ppm Test
Result
> 750
> 750
> 750
> 750
> 750
< 750
< 750
< 750
> 750
> 750
> 750
> 750
> 750
> 750
AGREEMENT3
Y, FP, FN
TRPH
Y
FP
FP
Y
Y
FN'
Y
Y
Y
Y
FP
FP
Y
Y
GC
Y
Y
Y
Y
Y
FN
FN
FN
Y
Y
FP
Y
Y
Y
Y Immunoassay and GC or IR results agree
FN False Negative
FP False Positive
FN* False Negative, but within 25% of GC or IR results
FP" False Positive, but within 25% of GC or IR results
4030-17
Revision 0
January 1995
-------
TABLE 12
IMMUNOASSAY COMPARED TO METHOD 8015
Field Trial 1: Gasoline (Interpretation at 100 ppm)
Sample ID
MW-18-1
MW-18-2
MW-18-3
MW-18-A1
MW-18-A1 Duplicate
MW-18-A2
DBS
MW-12-3
MW-13-1
MW-13-3
MW-13-4
MW-17-3
MW-17-4
MW-17-5
MW-16-2
MW-16-2 Duplicate
MW-19-2
MW-19-3
MW-14-1
MW-17-A1
Method 8015 (ppm)
270
15
15
20
15
1500
300
250
40
50
20
250
180
180
11,500
11,500
10
70
280
560
Field Trial 2: JP-4 Jet Fuel
Sample ID
TB1 6.5-7.0
TB2 6.5-7.0
TB1 5.0-5.5
TB2 5.0-5.5
TBS 5.0-5.5
TBS 6.5-7.0
TB5 5.0-5.5
TB5 6.5-7.0
TB4 6.5-7.0
TB4 5.5-6.0
Method 8015 (ppm)
15,900
16,800
900
100
ND(<5)
29,500
5,000
2,000
19,000
5,900
Immunoassay
Negative
Negative
Negative
Negative
Negative
Positive
Positive
Positive
Negative
Negative
Negative
Posi ti ve
Positive
Positive
Positive
Positive
Negative
Negative
Positive
Positive
(Interpretation at
Immunoassay
Positive
Positive
Negative
Positive
Negative
Positive
Positive
Positive
Positive
Positive
Concurrence?
False Negative
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
1,000 ppm)
Concurrence?
YES
YES
YES
False Positive
YES
YES
YES
YES
YES
YES
4030-18
Revision 0
January 1995
-------
METHOD 4035
SOIL SCREENING FOR POLYNUCLEAR AROMATIC HYDROCARBONS BY IMMUNOASSAY
1.0 SCOPE AND APPLICATION
1.1 Method 4035 is a procedure for screening soils to determine when total
polynuclear aromatic hydrocarbons (PAHs) are present at concentrations above 1
mg/Kg. Method 4035 provides an estimate for the concentration of PAHs by
comparison with a PAH standard.
1.2 Using the test kit from which this method was developed, >95% of
samples confirmed to have concentrations of PAHs below detection limits will
produce a negative result in the 1 ppm test configuration.
1.3 The sensitivity of the test is influenced by the binding of the target
analyte to the antibodies used in the kit. The commercial PAH kit used for
evaluation of this method is most sensitive to the three (i.e., phenanthrene,
anthracene, fluorene) and four (i.e. benzo(a)anthracene, chrysene, fluoranthene,
pyrene) ring PAH compounds listed in Method 8310, and also recognizes most of the
five and six ring compounds listed.
1.4 The sensitivity of the test is influenced by the nature of the PAH
contamination and any degradation processes operating at a site. Although the
action level of the test may vary from site to site, the test should produce
internally consistent results at any given site.
1.5 In cases where the exact concentration of PAHs are required,
quantitative techniques (i.e., Method 8310, 8270, or 8100) should be used.
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.
2.0 SUMMARY OF METHOD
2.1 An accurately weighed sample is first extracted and the extract
filtered using a commercially available test kit. The sample extract and an
enzyme conjugate reagent are added to immobilized antibody. The enzyme conjugate
"competes" with the PAHs present in the sample for binding to the immobilized
anti-PAH antibody. The test is interpreted by comparing the response produced
by testing a sample to the response produced by testing standard(s)
simultaneously.
2.2 A portion of all samples in each analytical batch should be confirmed
using quantitative techniques.
3.0 INTERFERENCES
3.1 Chemically similar compounds and compounds which might be expected to
be found in conjunction with PAH contamination were tested to determine the
4035 - 1 Revision 0
January 1995
-------
concentration required to produce a positive result. These data are shown in
Tables 1 and 2.
3.2 The kit was optimized to respond to three and four ring PAHs. The
sensitivity of the test to individual PAHs is highly variable. Naphthalene,
dibenzo(a,h)anthracene, and benzo(g,h,i)perylene have 0.5 percent or less than
the reactivity of phenanthrene with the enzyme conjugate.
3.3 The alkyl-substituted PAHs, chlorinated aromatic compounds, and other
aromatic hydrocarbons, such as dibenzofuran, have been demonstrated to be cross-
reactive with the immobilized anti-PAH antibody. The presence of these compounds
in the sample may contribute to false positives.
4.0 APPARATUS AND MATERIALS
4.1 PAH RISc™ Soil Test (EnSys, Inc.), or equivalent. Each commercially
available test kit will supply or specify the apparatus and materials necessary
for successful completion of the test.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Soil samples may be contaminated, and should therefore be considered
hazardous and handled accordingly.
7.0 PROCEDURE
7.1 Method 4035 is intended for field or laboratory use.
7.2 Follow the manufacturer's instructions for the test being used. Those
test kits used must meet or exceed the performance indicated in Tables 3-7.
7.3 The action limit for each application must be within the operating
range of the kit used.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for the test kit being used for
quality control procedures specific to the test kit used. Additionally, guidance
provided in Chapter One should be followed.
4035 - 2 Revision 0
January 1995
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8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.3 Do not use test kits past their expiration date.
8.4 Do not use tubes or reagents designated for use with other kits.
8.5 Use the test kits within the specified storage temperature and
operating temperature limits.
9.0 METHOD PERFORMANCE
9.1 The extraction efficiency of a commercially available test kit was
tested (PAH RISc™ Test, EnSys Inc.) by spiking phenanthrene, benzo(a)anthracene
and benzo(a)pyrene into PAH negative soil matrices (PAH-116 and PAH-141 are field
samples). The soils were spiked using detection limits established for each
compound (see Table 1), extracted and determined by immunoassay. The results for
these 3-, 4- and 5-ring PAHs (Table 4) demonstrated that they were extracted with
good recovery and yielded the correct assay interpretation.
9.2 A single laboratory study was conducted with a commercially available
test kit (PAH RISc™ Test, EnSys Inc.), using 25 contaminated soil samples. Four
replicate determinations were made on each test sample and the data compared with
values obtained using HPLC Method 8310. Several analysts performed the
immunoassay analyses. The immunoassay data agreed in all cases with the external
HPLC data obtained (Table 5).
9.3 An additional single laboratory validation study on 30 randomly
selected, PAH-contaminated field samples from multiple sites was run by the USEPA
Region X Laboratory. Results are reported in Table 6 on an as found basis, and
reported in Table 7 normalized to phenanthrene, based on cross-reactivity data
(from Table 1). The false positive rate at the 1 ppm action level was 13% for
unnormalized results and 19% for normalized results based on 31 analyses. The
false negative rate at 1 ppm was 0 in both cases. At the 10 ppm action level,
the false positive rate was 19% unnormalized and 26% normalized. False negative
rates at 10 ppm were 6% unnormalized and 3% normalized.
9.4 The probabilities of generating false positive and false negative
results at an action level of 1 ppm are listed in Table 3.
10.0 REFERENCES
1. PAH-RISc™ Users Guide, EnSys Inc.
2. P. P. McDonald, R. E. Almond, J. P. Mapes, and S. B. Friedman, "PAH-RISc™
Soil Test - A Rapid, On-Site Screening Test for Polynuclear Aromatic
Hydrocarbons in Soil", J. of AOAC International (accepted for publication
document #92263)
3. R. P. Swift, J. R. Leavell, and C. W. Brandenburg, "Evaluation of the
EnSys PAH-RISc™ Test Kit", Proceedings, USEPA Ninth Annual Waste Testing
and Quality Assurance Symposium, 1993.
4035 - 3 Revision 0
January 1995
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Table 1: Cross-Reactivity of Method 8310 PAHs
Compound
2 Rinqs
Naphthalene
3 Rinqs
Acenaphthene
Acenaphthylene
Phenanthrene
Anthracene
Fluorene
4 Rinqs
Benzo(a)anthracene
Chrysene
Fluoranthene
Pyrene
5 Rinqs
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
D ibenzo( a, h) anthracene
6 Rinqs
Indeno(l,2,3-c,d)pyrene
Benzo(g,h, i)perylene
Concentration Giving
a Positive Result
(ppm Soil Equivalent)
200
8.1
7.5
1.0
0.81
1.5
1.6
1.2
1.4
3.5
4.6
9.4
8.3
>200
11
>200
Percent
Cross-Reactivity
0.5
12
13
100
123
67
64
84
73
29
22
11
12
<0.5
9.4
<0.5
4035
Revision 0
January 1995
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Table 2: Cross Reactivity of Other PAHs and Related Compounds
Compound
Other PAHs
1 -Methyl naphthylene
2-Methylnaphthylene
1-Chloronaphthylene
Halowax 1013
Halowax 1051
Dibenzofuran
Other Compounds
Benzene
Toluene
CCA
Phenol
Creosote
2,4,6-Trichlorobenzene
2,3,5,6-
Tetrachl orobenzene
Pentachl orobenzene
Pentachlorophenol
Bis(2-ethylhexyl)
phthalate
Aroclor 1254
Aroclor 1260
Concentration Giving
a Positive Result
(ppm, Soil Equivalent)
54
58
59
18
>200
14
>200
>200
>200
>200
5.4
>200
>200
>200
>200
>200
>200
>200
Percent
Cross-Reactivity
1.8
1.7
1.7
5.7
<0.5
7.2
<0.5
<0.5
<0.5
<0.5
18.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
Table 3: Probability of False Negative and False Positive Results
for PAHs at a 1 ppm Action Level
Spike Concentration
Phenanthrene (ppm)
0
0.4
0.8
1.0
Probability of False
Positive (Mean ± SD)
0% ± 0%
23% ± 17%
94% ± 13%
N/A
Probability of False
Negative (Mean ± SD)
N/A
N/A
N/A
0% + 0%
Results were obtained from spiking four different validation lots, using
3 operators, 12 matrices for a total of 201 determinations at each
concentration of phenanthrene.
N/A = No false positive possible above action limit.
No false negative possible below action limit.
4035 - 5
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January 1995
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Table 4: Spike Recovery of Phenanthrene, Benzo(a)anthracene and Benzo(a)pyrene
Compound
Blank
Blank
Phenanthrene
Phenanthrene
Phenanthrene
Phenanthrene
Phenanthrene
Phenanthrene
Benzo(a)anthracene
Benzo( a) anthracene
Benzo(a)anthracene
Benzo(a)anthracene
Benzo(a)pyrene
Benzo(a)pyrene
Benzo(a)pyrene
Spike
(ppm)
0
0
1
1
1
10
10
10
1.6
1.6
16
16
8.3
8.3
83
Soil
Wake
PAH-116
Wake
PAH-116
PAH-141
Wake
PAH-116
PAH-141
Wake
PAH-116
Wake
PAH-116
Wake
PAH-116
PAH-116
PAH RISC™
Results
<1
<1
1-10
1-10
1-10
>10
>10
>10
1-10
1-10
>10
>10
1-10
1-10
>10
4035 - 6
Revision 0
January 1995
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Table 5: Powerplant Field Samples (Soil) Evaluated by Immunoassay
Field Sample
Number
PAH-137
PAH-141
PAH- 118
PAH-136
PAH- 139
PAH-126
PAH-127
PAH-122
PAH-138
PAH-131
PAH-128
PAH-132
PAH- 112
PAH-140
PAH- 130
PAH-116
PAH-135
PAH-133
PAH-119
PAH-120
PAH-124
PAH-134
PAH-114
PAH-113
PAH-115
EnSys Method
Immunoassay (ppm)
>10
<1
1-10
>10
>10
1-10, >10
>10
>10
>10
>10
>10
>10
>10
>10
>10
<1
>10
>10
>10
>10
>10
>10
>10
>10
>10
Method 8310
HPLC (ppm)
<21
<21
<26
26
<28
<32
<33
<33
33
<34
<35
<43
<48
50
54
<61
71
<91
<100
<161
<167
182
<247
<294
<343
4035 - 7
Revision 0
January 1995
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Table 6: Total PAH Content of Region X Field Samples Using EnSys
PAH RISc™ Immunoassay Test Kit
Sample ID
PAH-1
PAH- 2
PAH -3
PAH -4
PAH- 5
PAH -6
PAH -7
PAH -8
PAH -9
PAH-10
PAH- 11
PAH- 12
PAH-12Dup
PAH- 13
PAH-14
PAH- 15
PAH- 16
PAH- 17
PAH- 18
PAH-19
PAH- 20
PAH-21
PAH- 2 2
PAH- 2 3
PAH-24
PAH-25
1 ppm Test
<1
*
*
*
*
*
*
*
>1
it
it
*
*
*
*
*
*
*
*
10 ppm Test
<10
*
*
*
*
*
*
*
>10
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
GC/MS
Lab Result
(ppm)'
0.2
12.2
16.0
0.0
0.5
8.7
148
182
4.4
0.2
0.0
85.4
85.4
28.5
0.3
0.6
0.0
1.8
3.4
6.7
0.9
43.2
72.8
1.3
0.3
0.4
False +/-
Eval @
1 ppm
+
+
+
+
Eval @
10 ppm
+
+
+
+
+
+
i
'Sum of all PAHs detected.
4035 - 8
Revision 0
January 1995
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Table 6: Total PAH Content of Region X Field Samples Using EnSys
,TM
PAH RISc'M Immunoassay Test Kit (Contd.)
Sample ID
PAH-26
PAH-27
PAH- 2 8
PAH-29
PAH- 30
1 ppm Test
<1
*
*
>1
*
10 ppm Test
<10
*
*
*
*
*
>10
GC/MS
Lab Result
(ppm)1
27.9
0.0
16.4
0.4
9.6
False +/-
Eval @
1 ppm
Eval @
10 ppm
_
-
Table 7: Total PAH Content of Region X Field Samples Using EnSys
PAH RISc™ Immunoassay Test Kit Normalized to Cross-Reactivity
Sample ID
PAH-1
PAH-2
PAH -3
PAH -4
PAH -5
PAH- 6
PAH -7
PAH -8
PAH -9
PAH-10
PAH- 11
PAH-12
PAH-12Dup
PAH- 13
1 ppm Test
<1
*
*
*
>1
*
*
*
*
10 ppm Test
<10
>10
*
A
*
*
*
*
*
*
*
*
*
GC/MS
Lab Result
(ppm)1
0.1
8. 1
9.0
0.0
0.2
5.2
56.9
73.2
0.1
0.0
0.0
47.3
47.3
11.5
False +/-
Eval @
1 ppm
+
+
+
Eval @
10 ppm
+
+
+
+
+
+
xSum of all PAHs detected.
4035 - 9
Revision 0
January 1995
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Table 7: Total PAH Content of Region X Field Samples Using EnSys
PAH RISc Immunoassay Test Kit Normalized to Cross-Reactivity (Contd.)
Sample ID
PAH- 14
PAH- 15
PAH- 16
PAH- 17
PAH-18
PAH- 19
PAH- 20
PAH-21
PAH-22
PAH-23
PAH- 2 4
PAH-25
PAH-26
PAH- 2 7
PAH-28
PAH-29
PAH- 30
1 ppm Test
<1
it
it
*
*
*
*
>1
*
*
*
*
*
*
*
10 ppm Test
<10
it
it
*
*
*
*
*
*
it
*
*
*
>10
*
A
*
*
6C/MS
Lab Result
(ppm) '
0.2
0.5
0.0
1.2
1.7
3.6
0.6
27.5
49.2
0.8
0.1
0.2
13.5
0.0
6.4
0.2
2.8
False +/-
Eval @
1 ppm
+
+
+
Eval @
10 ppm
+
+
_
'Sum of all PAHs detected.
4035 - 10
Revision 0
January 1995
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METHOD 4040
SOIL SCREENING FOR TOXAPHENE BY IMMUNOASSAY
1.0 SCOPE AND APPLICATION
1.1 Method 4040 is a procedure for screening soils to determine whether
toxaphene (CAS Registry 8001-35-2) is present at concentrations above 0.5 jug/g.
Method 4040 provides an estimate for the concentration of toxaphene by comparison
against standards.
1.2 In cases where the exact concentration of toxaphene is required,
additional techniques (i.e., gas chromatography (Method 8081) or gas
chromatography/mass spectrometry (Method 8270)) should be used.
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 Test kits are commercially available for this method. The
manufacturer's directions should be followed.
2.2 In general, the method is performed using an extract of a soil
sample. Filtered extracts may be stored cold, in the dark. An aliquot of the
extract and an enzyme-toxaphene conjugate reagent are added to immobilized
toxaphene antibody. The enzyme-toxaphene conjugate "competes" with toxaphene
present in the sample for binding to immobilized toxaphene antibody. The
enzyme-toxaphene conjugate bound to the toxaphene antibody then catalyzes a
colorless substrate to a colored product. The test is interpreted by comparing
the color produced by a sample to the response produced by a reference reaction.
3.0 INTERFERENCES
3.1 Compounds that are chemically similar may cause a positive test
(false positive) for toxaphene. The test kit used to develop this method was
evaluated for interferences, and found to be relatively insensitive to other
organochlorine pesticides (e.g., Lindane, DDT and DDE). The data.for the lower
limit of detection of these compounds are provided in Table 1. Consult the
information provided by the manufacturer of the kit used for additional
information regarding cross reactivity with other compounds.
3.2 Storage and use temperatures may modify the method performance.
Follow the manufacturer's directions for storage and use.
4040-1 Revision 0
January 1995
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4.0 APPARATUS AND MATERIALS
4.1 Immunoassay test kit: EnviroGard™ Toxaphene in Soil (Millipore,
Inc.), or equivalent. Each commercially available test kit will supply or
specify the apparatus and materials necessary for successful completion of the
test.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4. 1 .
6.2 Soils samples may be contaminated, and should therefore be considered
hazardous and handled accordingly.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance specifications indicated
in Tables 2-5.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for the test kit being used
for quality control procedures specific to the test kit used. Additionally,
guidance provided in Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.3 Do not use test kits past their expiration date.
8.4 Do not use tubes or reagents designated for use with other test kits.
8.5 Use the test kits within their specified storage temperature and
operating temperature limits.
8.6 Method 4040 is intended for field or laboratory use. The appropriate
level of quality assurance should accompany the application of this method to
document data quality.
4040-2 Revision 0
January 1995
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9.0 METHOD PERFORMANCE
9.1 A single laboratory study was conducted using five spiked soil
samples and 30 real-world soils contaminated with Toxaphene. Results using the
immunoassay kit were compared with results obtained using Method 3540/8081.
Table 2 presents the data from this study. These data show a positive bias in
the immunoassay of almost 18%, compared to a 14% negative bias in Method
3540/8081.
9.2 Table 3 provides the results of single determinations in soil from
New Mexico.
9.3 Tables 8 and 9 provide data on the precision of Method 4040.
10.0 REFERENCES
1. EnviroGard™ Toxaphene Soil Test Kit Guide, Millipore, Inc.
2. Marsden, P.J., S-F Tsang, V. Frank, N. Chau, and M. Roby "Comparison of the
Millipore Immunoassay for Toxaphene with Soxhlet Extraction and Method 8081",
Science Applications International Corporation, under contract to Millipore
Inc., May 1994, unpublished.
4040-3 Revision 0
January 1995
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TABLE 1. POSSIBLE SOIL INTERFERENCES (a)
Compound
Diesel fuel
Endrin
Endosulfan I
Endosulfan II
Dieldrin
Heptachlor
Aldrin
technical Chlordane
gamma-BHC (Lindane)
alpha-BHC
delta-BHC
Soil Equivalent Concentrat
(ppb) Required to Yield a
Result
ion /xg/kg
Positive
45000
6
6
6
6
6
20
14
300
1000
1000
The following compounds were found to yield a negative result for concentrations
up to 200,000 jug/kg:
Gasoline
Pentachlorophenol
DDT
PCB (Aroclor 1248)
Trinitrotoluene
(a) Millipore, Inc. product literature
Table 2
Comparison of Method 4040 and Method 3540/8081
Spike
Concentration
(M9/9)
0.25
0.50
1.0
2.5
5.0
EnviroGard™ Toxaphene in Soil
Results
(/*g/g)
0.27
0.66
1.02
2.8
6.7
Average
% Recovery
108
132
102
112
134
117.6
Method 3540/Method 8081
Results
(/-tg/g)
0.19
0.33
0.83
2.9
5.5
% Recovery
75
66
84
116
110
L 85.6
4040-4
Revision 0
January 1995
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Table 3.
Results for New Mexico Soil Samples
Sample ID |f Toxaphene (jug/g)
Lab 1
Ml
M2
M3
M4
M5*
M6*
M7
LMS12
M8*
M9
M10
Mil*
M12*
M13*
M14
M15*
M16
M16-MS
M16-MSD
M17
Lab 2 II Method 3540/8081
II (Lab 1)
28,89
70,104
54,89
103,50
10,30
45,33
Nazilini
soil #12
0,33
23,104
78,33
64,5
53,75
33,75
17,75
65,33
82,75
82,75
82,75
19,50
0.09 J
0.04 J
0.04 J
0.01 J
40
19.3
<0.50
<0.50
0.26 J
1.0
0.14 J
0.27 J
27.2
0.14 J
0.48 J
0.21 J
6.0
6.0
4.8
Method 4040
(Lab 2)
0.3
0.9
0.8
0.2
58
54.8
0.2
1.7
1.1
2.6
2.1
11
42
0.9
2.8
1.8
NA
NA
L_ 6
AGREEMENT3
Y, FN, FP
FP
FP
FP
FP
Y
FP
Y
FP
FP
FP
FP
FP
Y
FP
FP
FP
-
-
Y
4040-5
Revision 0
January 1995
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Table 3. (continued)
Sample ID
Lab 1
M18
M19
M20
M21
M22
M23*
M24*
M24-MS
M24-MSD
M25*
M26
M27
M28
M29
M30*
M31
Lab 2
97,104
48,104
0,50
102,75
84,50
25,33
0,75
0,75
0,75
12,40 pit
0,89
0,104
98,89
104,33
76,89
40,50
Toxaphene (/j.g/g)
Method 3540/8081
(Lab 1)
0.049 J
0.054 J
1.3
0.15 J
0.058 J
89.6
0.5
3.7
3.6
35.6
0.16 J
0.88
0.41 J
0.30 J
0.10 J
323
Method 4040
(Lab 2)
0.6
1.1
2.3
0.3
0.9
130
1.9
NA
NA
45.5
6.9
2.1
3.4
0.7
5.8
460
AGREEMENT3
Y, FN, FP
FP
FP
Y
Y
FP
Y
FP
-
-
Y
FP
FP
FP
FP
FP
Y
NA = not analyzed
J = an estimate value. This is used to indicate the result is less than the
sample quantitation limit but greater than zero.
* DDE identified using GC/MS analyses
4040-6
Revision 0
January 1995
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Table 4
Optical Measurement Precision for Spiked Samples
Spike Level
(ppm)
0.40
4.0
0.40
4.0
0.40
4.0
Mean O.D.
(450 nm)
0.798
0.450
0.753
0.397
0.713
0.385
Percent RSD
5.6
9.7
9.0
4.7
6.6
7.8
n
8
8
8
8
8
L_ 8
O.D.
(2 ppm spike)
0.540
0.540
0.501
0.501
0.541
0.541
Table 5
Recovery and Precision of Three Types of Spiked Soils
Spike Cone, (ppm)
1.0
10.0
1.0
10.0
1.0
10.0
Mean percent
recovery
91.1
96.9
84.1
89.4
122.4
101.7
Percent RSD
20.0
10.4
14.6
4.2
8.8
2.0
n
3
3
3
3
3
3
overall percent recovery (n = 9), 1 ppm = 99 + 16%
overall percent recovery (n = 9), 10 ppm= 96 + 5%
4040-7
Revision 0
January 1995
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METHOD 4041
SOIL SCREENING FOR CHLORDANE BY IMMUNOASSAY
1.0 SCOPE AND APPLICATION
1.1 Method 4041 is a procedure for screening soils to determine whether
chlordane (CAS Registry 57-74-9) is present at concentrations above 20, 100 or
600 jug/kg. Method 4041 provides an estimate for the concentration of chlordane
by comparison against standards.
1.2 In cases where the exact concentration of chlordane is required,
additional techniques [i.e., gas chromatography (Method 8081) or gas
chromatography/mass spectrometry (Method 8270)] should be used.
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 Test kits are commercially available for this method. The
manufacturer's directions should be followed.
2.2 In general, the method is performed using an extract of a soil
sample. Filtered extracts may be stored cold, in the dark. An aliquot of the
extract and an enzyme-chlordane conjugate reagent are added to immobilized
chlordane antibody. The enzyme-chlordane conjugate "competes" with chlordane
present in the sample for binding to chlordane antibody. The enzyme-chlordane
conjugate bound to the chlordane antibody then catalyzes a colorless substrate
to a colored product. The test is interpreted by comparing the color produced
by a sample to the response produced by a reference reaction.
3.0 INTERFERENCES
3.1 Compounds that are chemically similar may cause a positive test
(false positive) for chlordane. The test kit used to develop this method was
evaluated for interferences. The data for the lower limit of detection of these
compounds are provided in Table 1. Consult the information provided by the
manufacturer of the kit used for additional information regarding cross
reactivity with other compounds.
3.2 Storage and use temperatures may modify the method performance.
Follow the manufacturer's directions for storage and use.
4041-1 Revision 0
January 1995
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4.0 APPARATUS AND MATERIALS
4.1 Immunoassay test kit: EnviroGard™ Chlordane in Soil (Millipore,
Inc.), or equivalent. Each commercially available test kit will supply or
specify the apparatus and materials necessary for successful completion of the
test.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Soil samples may be contaminated, and should therefore be considered
hazardous and handled accordingly.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance specifications indicated
in Tables 2-5.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for the test kit being used
for quality control procedures specific to the test kit used. Additionally,
guidance provided in Method 4000 and Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.3 Do not use test kits past their expiration date.
8.4 Do not use tubes or reagents designated for use with other test kits.
8.5 Use the test kits within their specified storage temperature and
operating temperature limits.
8.6 Method 4041 is intended for field or laboratory use. The appropriate
level of quality assurance should accompany the application of this method to
document data quality.
4041-2 Revision 0
January 1995
-------
9.0 METHOD PERFORMANCE
9.1 Method sensitivity was determined by establishing the "noise" level
expected from matrix effects encountered in negative soil samples and determining
the corresponding Chlordane concentration by comparison to the analyte-specific
response curve. 8 different soils which did not contain Chlordane were assayed.
Each of these soils was extracted in triplicate and each extract was assayed in
three different assays. The mean and the standard deviation of the resulting
%Bo's (%Bo = [(ODsample/ODnegatlvecontrol)xlOO]) was calculated and the sensitivity was
estimated at two standard deviations below the mean. The sensitivity for Method
4041 was determined to be 80% Bo at a 95% confidence interval. Based on the
average assay response to Chlordane, this corresponds to 6.4 ^g/kg Chlordane.
These data are shown in Table 2.
9.2 The effect of water content of the soil samples was determined by
assaying three different soil samples which had been dried and subsequently had
water added to 30% (w/w). Aliquots of these samples were then fortified with
Chlordane (100 ^g/kg). Each soil sample was assayed three times, with and
without added water, and with and without Chlordane fortification. It was
determined that water in soil up to 30% had no detectable effect on the method.
These data are shown in Table 3.
9.3 The effect of the pH of the soil extract was determined by adjusting
the soil pH of three soil samples. Soil samples were adjusted to pH 2 - 4 using
6N HC1 and pH 10 - 12 using 6N NaOH. Aliquots of the pH adjusted soil samples
were fortified with Chlordane (100 ^g/kg). Each soil sample was assayed
unadjusted and with pH adjusted to 2-4 and 10-12, both unfortified and fortified.
It was determined that soil samples with pH ranging from 3 to 11 had no
detectable effect on the performance of the method. These data are shown in
Table 4.
9.4 A field trial was undertaken to evaluate to ability of the
EnviroGard™ Chlordane in Soil Test Kit to identify chlordane contaminated soil
at a remediation site. A total of 32 soil samples were evaluated by both Method
4041 and Method 8080. Interpretation of the results at a 1 mg/kg cutoff resulted
in 2/32 (6.3%) false negatives and 0/32 (0%) false positives. Interpretation of
the results at a cutoff of 10 mg/kg resulted in 0/32 (0%) false negatives and
2/32 (6.3%) false positives. These data are shown in Table 5.
10.0 REFERENCES
1. EnviroGard™ Chlordane in Soil Test Kit Guide, Millipore, Inc.
4041-3 Revision 0
January 1995
-------
TABLE 1. CROSS REACTIVITY (a)
Compound
Chlordane
Endrin
Endosulfan I
Endosulfan II
Dieldrin
Heptachlor
Aldrin
Toxaphene
gamma-BHC (Lindane)
alpha-BHC
delta-BHC
Concentration Required for
Positive Interpretation (/*g/kg)
5
3
3
3
3
3
10
100
300
1000
1000
The following compounds were found to yield a negative result for
concentrations up to 200,000 jug/kg:
Gasoline PCB (Aroclor 1248)
Pentachlorophenol Trinitrotol uene
i
4041-4
Revision 0
January 1995
-------
AVERAGE
TABLE 2
METHOD SENSITIVITY
Soil#
SI
S2
S3
S4
S5
S6
S7
S8
Part 1
Soil Type
LOAM/SAND
LOAM
CLAY
CLAY
CLAY
LOAM/SAND
SAND
SAND
- Average Response with
Average %Bo (n = 8)
92.8
86.2
85.5
95.4
83.9
88.5
81.4
95.8
Negative Soils
Standard Deviation
2.0
1.0
8.8
1.1
2.6
1.8
2.7
0.8
88.7
4.5
Part 2 - Average Response with Chlordane Calibrators
Chlordane
Concentration
Average Absorbance Average %Bo
0
5
25
125
500
1.043
0.882
0.598
0.322
0.159
N/A
84.4
57.2
30.8
15.2
Part 3 - Method Sensitivity
Based on Part 1 and Part 2 Above:
Average %Bo - 2 SD = 79.7 which is equivalent to 6.4 ^g/kg Chlordane
Average %Bo - 3 SD = 75.2 which is equivalent to 8.6 ^g/kg Chlordane
4041-5
Revision 0
January 1995
-------
TABLE 3
EFFECT OF WATER CONTENT IN SOIL SAMPLES
Soil a
si
si
SI
SI
S2
S2
S2
S2
S3
S3
S3
S3
', Water
0
30
0
30
0
30
0
30
0
30
0
30
Fortified? Reo. 1
No
No
Yes
Yes
No
No
Yes
Yes
No
No
Yes
Yes
95.2
96.0
40.5
42.2
85.8
78.7
37.7
39.8
76.6
87.4
40.0
40.8
Reo. 2
101
99.2
38.5
43.0
87.1
84.9
39.5
38.8
76.6
88.7
40.2
37.1
Rep. 3
94.5
96.0
35.9
43.0
85.5
79.8
40.6
37.0
73.0
85.7
35.7
38.7
Mean Std. Dev.
97.0
97.1
38.3
42.8
86.1
81.1
39.3
38.5
75.4
87.3
38.7
38.9
3.7
1.8
2.3
0.5
0.9
3.3
1.5
1.4
2.1
1.5
2.5
1.9
± 2 SD Ranqe
89.6
93.5
33.7
41.8
84.3
74.5
36.3
35.7
71.2
84.3
33.7
35.1
- 104
- 101
- 42.9
- 43.8
- 87.9
- 87.8
- 42.3
- 41.3
- 79.6
- 90.3
-43.7
- 42.7
4041-6
Revision 0
January 1995
-------
TABLE 4
EFFECT OF pH OF SOIL SAMPLES
oil
SI
SI
SI
SI
SI
SI
S2
S2
S2
S2
S2
S2
S3
S3
S3
S3
S3
S3
pH Adj. Fortified? Rep
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
No
No
No
Yes
Yes
Yes
No
No
No
Yes
Yes
Yes
No
No
No
Yes
Yes
Yes
97
97
107
43
43
44
87
94
89
43
44
42
72
85
89
44
40
40
. 1* Rep. 2
.5
.4
.6
.6
.8
.8
.2
.3
.9
.6
.3
.3
.3
.4
.5
.5
.6
87.8
114
114
47.5
51.8
50.8
86.3
108
99.3
48.9
55.9
59.2
74.5
105
83.8
49.5
52.1
37.1
Rep. 3
94.8
94.7
93.8
38.5
34.1
32.0
87.6
80.5
77.9
33.9
41.5
36.5
79.3
75.7
85.9
32.6
34.7
43.9
Mean
93.4
102
105
43.2
43.2
42.5
87.3
94.1
88.8
42.2
47.4
46.0
75.4
88.8
86.4
42.2
42.4
40.5
Std.
5
10
10
4
8
9
0
13
10
7
7
11
3
15
2
8
8
3
Dev.
.0
.7
.1
.5
.8
.6
.8
.5
.7
.7
.6
.8
.6
.1
.8
.7
.9
.4
± 2 SD Range
83.4
80.8
84.7
34.2
25.6
23.3
85.7
67.1
67.4
26.8
32.2
22.4
68.2
58.6
80.8
24.8
24.6
33.7
- 103
- 124
- 125
- 52.2
- 60.8
- 61.7
- 88.9
~ 121
- 110
- 57.6
- 62.6
- 69.6
- 82.6
- 119
- 92.0
- 59.6
- 60.2
- 47.3
All values shown are %Bo [= (ODsample/ODnegatlve C0ntrol)xl00]
4041-7
Revision 0
January 1995
-------
Sample ID
co-ss-2
co-ss-3
co-ss-4
co-ss-5
co-ss-6
co-ss-7
co-ss-8
co-ss-9
co-ss-10
co-ss-13
co-ss-14
co-ss-15
co-ss-17
co-ss-20
co-ss-21
co-ss-22
co-ss-23
co-ss-24
co-ss-25
co-ss-26
co-ss-27
co-ss-28
co-ss-28-170
co-ss-29
co-ss-30
co-ss-31
co-ss-32
co-ss-33
co-ss-34
co-ss-35
co-ss-36
co-ss-41
TABLE 5
Correlation to Method 8081
Test Interpretation at 1 mg/kg
Method 8081 (mg/kg) Immunoassay (mg/kg)
45
4.9
25
1.4
2.7
2.5
<1.0
7.9
6.0
5.2
2.9
2.1
<1.0
2.8
51
1.4
9.6
2.6
14
1.8
2.9
4.2
5.9
POSITIVE
POSITIVE
POSITIVE
NEGATIVE
POSITIVE
POSITIVE
NEGATIVE
POSITIVE
POSITIVE
POSITIVE
POSITIVE
POSITIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
POSITIVE
POSITIVE
NEGATIVE
NEGATIVE
POSITIVE
POSITIVE
POSITIVE
POSITIVE
NEGATIVE
NEGATIVE
POSITIVE
POSITIVE
NEGATIVE
POSITIVE
NEGATIVE
Results Agree?
YES
YES
YES
FALSE NEGATIVE
YES
YES
YES
YES
YES
YES
YES
YES
YES
FALSE NEGATIVE
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
4041-8
Revision 0
January 1995
-------
TABLE 5 (continued)
Correlation to Method 8081
Test Interpretation at 10 mg/kg
Sample ID Method 8081 (mg/kg)
Immunoassay (mg/kg) Results Agree?
co-ss-2
co-ss-3
co-ss-4
co-ss-5
co-ss-6
co-ss-7
co-ss-8
co-ss-9
co-ss-10
co-ss-13
co-ss-14
co-ss-15
co-ss-17
co-ss-20
co-ss-21
co-ss-22
co-ss-23
co-ss-24
co-ss-25
co-ss-26
co-ss-27
co-ss-28
co-ss-28-170
co-ss-29
co-ss-30
co-ss-31
co-ss-32
co-ss-33
co-ss-34
co-ss-35
co-ss-36
co-ss-41
45
4.9
25
1.4
2.7
2.5
<1.0
7.9
6.0
5.2
2.9
2.1
<1.0
2.8
51
1.4
9.6
2.6
14
1.8
2.9
4.2
5.9
POSITIVE
NEGATIVE
POSITIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
POSITIVE
POSITIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
POSITIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
POSITIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
YES
YES
YES
YES
YES
YES
YES
FALSE POSITIVE
FALSE POSITIVE
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
4041-9
Revision 0
January 1995
-------
METHOD 4042
SOIL SCREENING FOR DDT BY IMMUNOASSAY
1.0 SCOPE AND APPLICATION
1.1 Method 4042 is a procedure for screening soils to determine whether
l,l,l-trichloro-2,2-bis (4-chlorophenyl) ethane (DDT) (CAS Registry 50-29-3) and
its breakdown products (ODD, DDE, and DDA) are present at concentrations above
0.2, 1.0 or 10 mg/kg. Method 4042 provides an estimate for the sum of
concentrations of DDT and daughter compounds by comparison against standards.
1.2 In cases where the exact concentration of DDT is required, additional
techniques [i.e., gas chromatography (Method 8081) or gas chromatography/mass
spectrometry (Method 8270)] should be used.
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 Test kits are commercially available for this method. The
manufacturer's directions should be followed.
2.2 In general, the method is performed using an extract of a soil
sample. Filtered extracts may be stored cold, in the dark. An aliquot of the
extract and an enzyme-DDT conjugate reagent are added to immobilized DDT
antibody. The enzyme-DDT conjugate "competes" with DDT present in the sample for
binding to DDT antibody. The enzyme-DDT conjugate bound to the DDT antibody then
catalyzes a colorless substrate to a colored product. The test is interpreted
by comparing the color produced by a sample to the response produced by a
reference reaction.
3.0 INTERFERENCES
3.1 Compounds that are chemically similar may cause a positive test
(false positive) for DDT. The test kit used to develop this method was evaluated
for interferences. The data for the lower limit of detection of these compounds
are provided in Table 1. Consult the information provided by the manufacturer
of the kit used for additional information regarding cross reactivity with other
compounds.
3.2 Storage and use temperatures may modify the method performance.
Follow the manufacturer's directions for storage and use.
4042-1 Revision 0
January 1995
-------
4.0 APPARATUS AND MATERIALS
4.1 Immunoassay test kit: EnviroGard™ DDT in Soil (Millipore, Inc.), or
equivalent. Each commercially available test kit will supply or specify the
apparatus and materials necessary for successful completion of the test.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Soil samples may be contaminated, and should therefore be considered
hazardous and handled accordingly.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance specifications indicated
in Tables 2-5.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for the test kit being used
for quality control procedures specific to the test kit used. Additionally,
guidance provided in Method 4000 and Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.3 Do not use test kits past their expiration date.
8.4 Do not use tubes or reagents designated for use with other test kits.
8.5 Use the test kits within their specified storage temperature and
operating temperature limits.
8.6 Method 4042 is intended for field or laboratory use. The appropriate
level of quality assurance should accompany the application of this method to
document data quality.
4042-2 Revision 0
January 1995
-------
9.0 METHOD PERFORMANCE
9.1 Method sensitivity was determined by establishing the "noise" level
expected from matrix effects encountered in negative soil samples and determining
the corresponding DDT concentration by comparison to the analyte-specific
response curve. 8 different soils which did not contain DDT were assayed. Each
of these soils was extracted in triplicate and each extract was assayed in three
different assays. The mean and the standard deviation of the resulting %Bo's
(%Bo = [(ODsample/ODnegativecontrol)xlOO]) were calculated and the sensitivity was
estimated at two standard deviations below the mean. The sensitivity for Method
4042 was determined to be 81.4% Bo at a 95% confidence inteval. Based on the
average assay response to DDT, this corresponds to 0.044 ppm DDT. These data are
shown in Table 2.
9.2 The effect of water content of the soil samples was determined by
assaying three different soil samples which had been dried and subsequently had
water added to 30% (w/w). Aliquots of these samples were then fortified with DDT
(1.0 mg/kg). Each soil sample was assayed three times, with and without added
water, and with and without DDT fortification. It was determined that water in
soil up to 30% had no detectable effect on the method. These data are shown in
Table 3.
9.3 The effect of the pH of the soil extract was determined by adjusting
the soil pH of three soil samples. Soil samples were adjusted to pH 2 - 4 using
6N HC1 and pH 10 - 12 using 6N NaOH. Aliquots of the pH adjusted soil samples
were fortified with DDT (1.0 mg/kg). Each soil sample was assayed unadjusted and
with pH adjusted to 2-4 and 10-12, both unfortified and fortified. It was
determined that soil samples with pH ranging from 3 to 11 had no detectable
effect on the performance of the method. These data are shown in Table 4.
9.4 A field study was conducted at a contaminated site using a
commercially available test kit (EnviroGard™ DDT in Soil Test Kit, Millipore
Corp.). The immunoassay was used to identify soil which had been contaminated
with DDT. The standard method (Method 8080) was performed at a certified
laboratory and the results were compared to the immunoassay. When interpreting
the results at a 0.2 ppm cutoff, the immunoassay yielded 0/32 (0%) false
negatives and 2/32 (6.3%) false positives. When interpreting the results at a
1.0 ppm cutoff, the immunoassay yielded 1/32 (3.1%) false negatives and 2/32
(6.3%) false positives. These data are shown in Table 5.
10.0 REFERENCES
1. EnviroGard™ DDT in Soil Test Kit Guide, Millipore, Inc.
4042-3 Revision 0
January 1995
-------
TABLE 1. CROSS REACTIVITY (a)
Compound
p,p'-DDT
p,p'-DDD
p,p'-DDE
o,p'-DDT
o,p'-DDD
o,p'-DDE
DDA
Chloropropylate
Chlorobenzilate
Dicofol
Chloroxuron
Monol inuron
Thiobencarb
Tebuconazole
Neburon
Tetradifon
Diclofop
PCB (Aroclor 1248)
Concentration Required for
Positive Interpretation (ppm)
0.04
0.01
0.18
4.0
0.4
3.0
0.002
0.007
0.03
0.14
24
25
5
7
17
1.2
70
90
The following analytes are not detected at or above 100 ppm:
2,4-D 4-chlorophenoxyacetic acid Chlordane
Pentachlorophenol Chlorbromuron Chlortoluron
Dicamba Diflubenzuron Diuron
Lindane Linuron MCPA acid
MCPB Mecoprop Gasoline
Diesel Fuel 2,4,6-Trinitrotoluene Toxaphene
i
4042-4
Revision 0
January 1995
-------
AVERAGE
TABLE 2
Method Sensitivity
Soil#
SI
S2
S3
S4
S5
S6
S7
S8
Part 1
Soil Type
LOAM
CLAY
SAND
LOAM
LOAM/SAND
CLAY
LOAM/SAND
SAND/ LOAM
- Average Response with
Average %Bo (n = 9}
87.0
93.2
97.2
87.7
88.1
100.8
103.6
89.6
Negative Soils
Standard Deviation
7.5
2.3
2.6
1.2
2.3
2.1
0.3
4.5
93.4
6.0
Part 2 - Average Response with DDT Calibrators
DDT
Concentration (ppm)
Average Absorbance
Average %Bo
0
0.1
1.0
10.0
50.0
1.133
0.897
0.569
0.362
0.259
N/A
79.4
50.3
32.0
22.9
Part 3 - Method Sensitivity
Based on Part 1 and Part 2 Above:
Average %Bo - 2 SD = 81.4 which is equivalent to 0.044 ppm DDT
Average %Bo - 3 SD = 75.4 which is equivalent to 0.097 ppm DDT
4042-5
Revision 0
January 1995
-------
TABLE 3
EFFECT OF WATER CONTENT IN SOIL SAMPLES
SI
SI
SI
SI
S2
S2
S2
S2
S3
S3
S3
S3
0
30
0
30
0
30
0
30
0
30
0
30
•tified?
No
No
Yes
Yes
No
No
Yes
Yes
No
No
Yes
Yes
Rep. 1
79.7*
89.1
49.8
55.8
85.2
94.8
54.4
56.3
96.2
95.6
54.8
59.4
Rep
79
84
62
59
96
94
47
53
91
90
52
55
. 2
.3
.0
.1
.9
.2
.3
.0
.8
.3
.5
.9
.0
Rep
83
85
46
58
97
95
56
60
100
96
54
54
. 3
.7
.9
.3
.0
.9
.0
.1
.2
.0
.4
.8
.5
Mean
80.9
86.4
52.8
57.9
93.1
94.7
52.5
56.8
95.8
94.2
54.2
56.3
Std.
2
2
8
2
6
0
4
3
4
3
1
2
Dev.
.4
.6
.3
.1
.9
.3
.8
.2
.3
.2
.1
.7
± 2
76.
81.
36.
53.
79.
94.
42.
50.
87.
87.
52.
50.
SD
1 -
2 -
2 -
7 -
3 -
1 -
9 -
4 -
2 -
8 -
0 -
9 -
Range
85.7
91.6
69.4
62.1
106.9
95.3
62.1
63.2
104.4
100.6
56.4
61.7
* All values shown are %Bo [= (ODsample/ODn
egative conti
4042-6
Revision 0
January 1995
-------
TABLE 4
EFFECT OF pH OF SOIL SAMPLES
oil
SI
SI
SI
SI
SI
SI
S2
S2
S2
S2
S2
S2
S3
S3
S3
S3
S3
S3
pH Adj. Fortified? Rep
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
None
Acidic
Basic
No
No
No
Yes
Yes
Yes
No
No
No
Yes
Yes
Yes
No
No
No
Yes
Yes
Yes
91
79
80
57
54
52
94
87
87
51
52
52
99
86
94
56
54
54
. 1* Rep. 2
.4
.7
.5
.5
.2
.8
.7
.8
.9
.7
.2
.0
.1
.4
.9
.2
.5
.6
91.3
87.0
84.5
60.3
60.6
60.2
90.6
100.1
81.6
56.9
61.0
53.5
94.2
84.3
100.3
54.3
53.5
57.2
Rep. 3
78.3
86.8
78.5
55.1
55.2
53.3
94.5
100.9
98.3
48.3
55.2
48.9
98.2
85.5
92.9
52.8
53.9
62.9
Mean Std.
87.0
84.5
81.2
57.6
56.7
55.5
93.2
96.3
89.3
52.3
56.1
51.5
97.2
85.4
96.1
54.4
54.0
58.2
7
4
3
2
3
4
2
7
8
4
4
2
2
1
3
1
0
4
Dev
.5
.1
.0
.6
.4
.1
.3
.3
.5
.3
.5
.3
.6
.1
.8
.7
.5
.2
. ± 2 SD Range
72.0
76.3
75.2
52.4
49.9
47.3
88.6
81.7
72.3
43.7
47.1
46.9
92.0
83.2
88.5
51.0
53.0
49.8
- 102
- 92.7
- 87.2
- 62.8
-63.5
- 63.7
- 97.8
- Ill
- 106
- 60.9
- 65.1
- 56.1
- 102
- 87.6
- 104
- 57.8
- 55.0
- 66.6
* All values shown are %Bo [= (OD5amp,yODnegatlvecontrol)xlOO]
4042-7 Revision 0
January 1995
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TABLE 5
Comparison to Method 8081
Test Interpretation at 0.2 mg/kg
Sample ID
co-ss-2
co-ss-3
co-ss-4
co-ss-5
co-ss-6
co-ss-7
co-ss-8
co-ss-9
co-ss-10
co-ss-13
co-ss-14
co-ss-15
co-ss-17
co-ss-20
co-ss-21
co-ss-22
co-ss-23
co-ss-24
co-ss-25
co-ss-26
co-ss-27
co-ss-28
co-ss-28-170
co-ss-29
co-ss-30
co-ss-31
co-ss-32
co-ss-33
co-ss-34
co-ss-35
co-ss-36
co-ss-41
Method 8081 (mg/kg)
3.6
0.55
2.3
<0.05
0.15
0.3
0.1
0.8
0.23
0.79
0.58
0.35
<0.05
0.18
0.06
<0.05
<0.05
1.2
0.12
<0.05
<0.05
0.16
0.18
0.69
0.73
0.68
<0.05
0.32
0.23
0.52
1.0
<0.05
Immunoassay (mq/kg)
POSITIVE
POSITIVE
POSITIVE
NEGATIVE
POSITIVE
POSITIVE
NEGATIVE
POSITIVE
POSITIVE
POSITIVE
POSITIVE
POSITIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
POSITIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
POSITIVE
POSITIVE
POSITIVE
POSITIVE
NEGATIVE
POSITIVE
POSITIVE
POSITIVE
POSITIVE
NEGATIVE
Results Agree?
YES
YES
YES
YES
FALSE POSITIVE
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
FALSE POSITIVE
YES
YES
YES
YES
YES
YES
YES
YES
YES
4042-8
Revision 0
January 1995
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TABLE 5 (continued)
Comparison to Method 8081
Test Interpretation at 1.0 mg/kg
Sample ID
co-ss-2
co-ss-3
co-ss-4
co-ss-5
co-ss-6
co-ss-7
co-ss-8
co-ss-9
co-ss-10
co-ss-13
co-ss-14
co-ss-15
co-ss-17
co-ss-20
co-ss-21
co-ss-22
co-ss-23
co-ss-24
co-ss-25
co-ss-26
co-ss-27
co-ss-28
co-ss-28-170
co-ss-29
co-ss-30
co-ss-31
co-ss-32
co-ss-33
co-ss-34
co-ss-35
co-ss-36
co-ss-41
Method 8081 (mg/kg)
3.6
0.55
2.3
<0.05
0.15
0.3
0.1
0.8
0.23
0.79
0.58
0.35
<0.05
0.18
0.06
<0.05
<0.05
1.2
0.12
<0.05
<0.05
0.16
0.18
0.69
0.73
0.68
<0.05
0.32
0.23
0.52
1.0
<0.05
Immunoassay (mg/kg)
POSITIVE
NEGATIVE
POSITIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
POSITIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
POSITIVE
POSITIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
NEGATIVE
Results Agree?
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
FALSE POSITIVE
FALSE POSITIVE
YES
YES
YES
YES
FALSE NEGATIVE
YES
4042-9
Revision 0
January 1995
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METHOD 4050
TNT EXPLOSIVES IN WATER AND SOILS BY IMMIJNOASSAY
1.0 SCOPE AND APPLICATION
1.1 Method 4050 is a procedure for screening waters and soils to
determine when trinitrotoluene (TNT, CAS No. 118-96-7) is present at
concentrations above 0.5 mg/kg in soil and 5 jiig/L in water. Method 4050
provides an estimate for the concentration of TNT by comparison with a reference.
1.2 Using the test kit from which this method was developed, 93% of soil
samples containing 0.25 ppm or less of TNT will produce a negative result, and
99+% of soil samples containing 1.0 ppm or greater of TNT will produce a positive
result. In addition, 93% of water samples containing 2.5 ppb or less of TNT will
produce a negative result, and 99%+ of water samples containing 10 ppb or more
of TNT will produce a positive result.
1.3 In cases where the exact concentrations of TNT are required,
quantitative techniques (i.e., Method 8330) should be used.
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 Test kits are commercially available
manufacturer's directions should be followed.
for this method.
The
2.2 In general, the method is performed using a diluted water sample or
an extract of a soil sample. Samples and an enzyme-TNT conjugate reagent are
added to immobilized TNT antibody. The enzyme-TNT conjugate "competes" with TNT
present in the sample for binding to immobilized TNT antibody. The enzyme-TNT
conjugate bound to the TNT antibody then catalyzes a colorless substrate to a
colored product. The test is interpreted by comparing the color produced by a
sample to the response produced by a reference reaction.
3.0 INTERFERENCES
3.1 Chemically similar compounds and compounds that might be expected to
be found in conjunction with TNT contamination were tested to determine the
concentration required to produce a positive test result.
3.1.1 Table 1 provides the concentrations of compounds tested with
the D TECH test kit that are required to elicit a positive response at the
MDL, as well as the concentration required to yield 50% inhibition
4050-1
Revision 0
January 1995
-------
compared to the standard curve.
4.0 APPARATUS AND MATERIALS
4.1 Immunoassay test kit: D TECH™ TNT (Strategic Diagnostics Inc.), or
equivalent. Each commercially available test kit will supply or specify the
apparatus and materials necessary for successful completion of the test.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary,, for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND TRANSPORTATION
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Soil samples may be contaminated, and should therefore be considered
hazardous and handled accordingly.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance specifications indicated
in Tables 3-6.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for the test kit being used
for quality control procedures specific to the test kit used. Additionally,
guidance provided in Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.3 Do not use test kits past their expiration date.
8.4 Do not use tubes or reagents designated for use with other test kits.
Do not mix reagents from one kit lot with a different kit lot.
8.5 Use the test kits within their specified storage temperature and
operating temperature limits.
4050-2 Revision 0
January 1995
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8.6 Method 4050 is intended for field or laboratory use. The appropriate
level of quality assurance should accompany the application of this method to
document data quality.
9.0 METHOD PERFORMANCE
9.1 Table 1 provides data on the minimum concentrations of possible
interferants and co-contaminants required to elicit a positive response in the
test kits evaluated.
9.2 Twenty five soil samples, known to not be contaminated with TNT, were
extracted and analyzed using the DTECH TNT kit to determine the extent of soil
matrix effects on the performance of the test kit. The results are provided in
Table 2A, and show that false positive results are not attributable to soil
components. Table 2B presents similar data generated from the analysis of thirty
uncontaminated water samples.
9.3 Thirty water samples and thirty soil samples, known to not be
contaminated with TNT, were each spiked with TNT at one-half and two times the
MDL (0.25 and 1.0 ppm respectively). These samples were analyzed with the DTECH
TNT test kit to determine the error rate of the assay. The results are presented
in Tables 3A and 3B.
9.4 Ten different soil types, all known to not be contaminated with TNT,
were spiked with an acetone solution containing approximately 1.0 ppm TNT. This
spiking solution was later quantitated by Method 8330 and found to contain 0.77
ppm TNT. The spiked soil samples were analyzed three (3) times with the DTECH
kit to determine the extraction efficiency of the method. The data are presented
in Table 4.
9.5 Table 5 presents the results of analysis of three soils spiked at
approximately 1 and 3 ppm TNT. Each sample was analyzed once using Method 8330
and ten times using the DTECH kit.
9.6 Tables 6A and 6B present the results of two field trials. In each
trial, soil samples were obtained at a West Coast site from borings, using a
split spoon technique. The samples were homogenized by placing approximately six
cubic inches of soil into a stainless steel vessel and mixing for five minutes
with a stainless steel trowel. The soil was aliquotted into two (2) six ounce
glass bottles, tested on-site using the DTECH method and transported to
commercial laboratories (one laboratory per field trial) for analysis by Method
8330. Table 6C presents the results of a third party field trial, conducted by
the California Department of Environmental Health Services.
10.0 REFERENCES
1. D TECH™ TNT Users Guide , SDI/EM Sciences 1994
4050-3 Revision 0
January 1995
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2. Mutter,L., G. Teaney, and J.W.Stave, "A Novel Field Screening System for
TNT Using EIA", in Field Screening Methods for Hazardous Wastes and
Toxic Chemicals, Vol 1, Proceedings of the 1993 U.S. EPA/A&WMA
International Symposium, p.472, 1993.
3. Teaney, G., J.Melby, L.Mutter and J.Stave, "A Novel Field Analytical
Method for TNT", Proceedings of the American Association of Analytical
Chemists, 1993.
4. Haas, R.J., and B.P. Simmons, "Measurement of Trinitrotoluene (TNT) and
Hexahydro-l,3,5-trinitro-l,3,5-triazine (RDX) in Soil by Enzyme
Immunoassay and High Performance Liquid Chromatography (EPA Method
8330)", California Environmental Protection Agency, Department of Toxic
Substances Control, Hazardous Materials Laboratory, March, 1995.
4050-4 Revision 0
January 1995
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TABLE 1
CROSS REACTANTS
D TECH™ TNT test kit
COMPOUND
TNT (2,4,6-trinitrotoluene)
Tetryld
1, 3, 5-tri nitrobenzene
2-amino-4,6-dinitrotoluene
4-amino-2,6-dinitrotoluene
2,4-dinitrotoluene
2,6-diaminonitrotoluene
2-nitrophenol
4-nitrophenol
2,4-dinitrophenol
RDXd
HMXd
MDLa
(ppm)
0.5
3
4
13
>500
90
>500
>500
>500
>500
>500
>500
1C 50b
(ppm)
17
48
75
150
>500
390
>500
>500
>500
>500
>500
>500
% CROSS
REACTIVITY0
100
35
23
11
<1
4
<1
<1
<1
<1
<1
<1
The following compounds were not detected at or above 100 ppm:
Benzene Xylenes PCB 1254 Triazine
Ethyl benzene Toluene PCP
PAHs - an equal concentration mixture of:
Acenaphthene Acenaphthalene Anthracene
1,2-Benzanthracene Benzo(a)pyrene Benzo(b)fluoranthene
Benzo(ghi )perylene Benzo(k)fl uoranthene Chrysene
Dibenz(ah)anthracene Fluoranthene Fluorene
Indeno(123-cd)pyrene Naphthalene Phenanthrene
Pyrene
The Method Detection Limit (MDL) is defined as the lowest concentration
of compound that yields a positive test result.
The IC50 is defined as the concentration of compound required to produce
a test response equivalent to 50% of the maximum response.
% Cross reactivity is determined by dividing the equivalent TNT
concentration by the actual compound concentration at IC50
Tetryl = methyl-2,4,6-trinitrophenylnitramine
RDX = hexahydro-l,3,5-trinitro-l,3,5-triazine
HMX = octahydro-l,3,5,7-tetranitro-l,3,5,7-tetrazocine
4050-5
Revision 0
January 1995
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TABLE 2A
SOIL MATRIX EFFECTS
Soil
133
101
100
102
106
107
109
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
127
128
130
Soil type
Avonburg Fine Sine Silt
Matapeake Silt Loam
Clay Loam
Sassafras Sand Loam
Evesboro Low Organic Sand
Pokomoke High OM Sand
Davidson Clay Loam
Shontic Casa Grande Sand
Casa Grande Clay Loam
Trix Sand Clay Loam
Trix Casa Grande Clay
Yolo Loam
Capay Silt Clay
Sycamore Silt Loam
Dennis Silt Loam
Grundy Silt Clay Loam
Luray Silt Clay Loam
Wooster Silt Loam
Vienna Loam
Opal Clay
Raub Silt Loam
Rockfield Silt Loam
Ci sne
Muscatine Loam
Sandy Brae
N/A
DE
DE
DE
DE
DE
GA
AZ
AZ
AZ
AZ
CA
CA
CA
KS
KS
OH
OH
SO
SD
IN
IN
IL
IL
DE
D TECH
RANGE (ppm)
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
4050-6
Revision 0
January 1995
-------
TABLE 2B
WATER MATRIX EFFECTS
Water
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
Water type
Ground Water, Burlington
Well Water, Burlington
Surface Water #1, Houston
Unknown Creek, Dartmouth
City Well Water, Ontario
Pacific Ocean, Victoria
Surface Water, Harmony Woods
Adamsville River, Adamsville
Surface Water #2, Houston
Buttermilk Falls, White Haven
Main St Pond, Germantown
Hudson River, Germantown
Atlantic Ocean
Ground Water #1, Dover
Ground Water #2, Dover
Ground Water #3, Dover
Drinking Well Water,
Ground Water, Elsmere
Ground Water, Elsmere
Ground Water, Elsmere
Lab Sample 20643
Lab Sample 20645
Lab Sample 20659
Lab Sample 20826
Lab Sample 20827
Lab Sample 20843
Lab Sample 20850
Lab Sample 20848
Ground Water, Adrian
Ground Water, Adrian
IA
IA
TX
MA
CA
CA
DE
RI
TX
PA
NY
NY
NJ
DE
DE
DE
PA
DE
DE
DE
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
GA
GA
D TECH RANGE (ppm)
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
<5.0
4050-7
Revision 0
January 1995
-------
TABLE 3A
False Negative and False Positive Rates, Soil Matrix5
Spike Concentration
0.25 ppm
1 .0 ppm
False Positive Rate
7%
-
False Negative Rate
_
0%
a Thirty negative soils were spiked with TNT at one-half and two times the MDL
(0.25 and 1.0 ppm respectively). These samples were analyzed with the DTECH TNT
test kit to determine the error rate of the assay.
TABLE 3B
False Negative and False Positive Rates, Water Matrix6
i
Spike Concentration
0.25 ppm
1.0 ppm
False Positive Rate
7%
100%
False Negative Rate
93%
0%
a Thirty negative water samples were spiked with TNT at one-half and two times
the MDL (0.25 and 1.0 ppm respectively). These samples were analyzed with the
DTECH TNT test kit to determine the error rate of the assay.
4050-8
Revision 0
January 1995
-------
TABLE 4
DETERMINATION OF EXTRACTION EFFICIENCY FROM SOIL SAMPLES3
SOIL ID
101
106
108
109
110
116
117
123
126
128
SPIKING
SOLUTION
MEAN TNT CONC.
(ppm)
0.54
0.64
0.87
0.63
0.88
1.02
0.82
0.87
0.95
0.65
0.77
SD
0.04
0.06
0.18
0.08
0.15
0.15
0.15
0.23
0.26
0.11
N/A
%CV
7
9
20
13
17
17
15
26
28
16
N/A
%RECOVERY
70
84
113
82
115
115
132
113
123
84
100
aTen different TNT negative soils were spiked with an acetone solution containing
0.77 TNT. The spiked soil samples were analyzed three times with the DTECH kit
to determine the extraction efficiency of the method.
4050-9
Revision 0
January 1995
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TABLE 5
RECOVERY OF TNT SPIKED INTO REAL SOILS
Three (3) soils were spiked at approximately 1 and 3 ppm TNT.
analyzed once by Method 8330 and ten (10) times by D TECH.
Each sample was
SAMPLE ID
106-1
116-1
128-1
AMOUNT SPIKED
1.0
1.0
1.0
D TECH (ppm)
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
1.5 - 3.0
0.5 - 1.5
0.5 - 1.5
1.5 - 3.0
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
1.5 - 3.0
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
1.5 - 3.0
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
HPLC METHOD
8330
0.69
0.73
0.75
AGREEMENT8
Y, FN, FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
FP
Y
Y
FP
Y
Y
Y
Y
Y
FP
Y
Y
Y
Y
Y
FP
Y
Y
Y
Y
4050-10
Revision 0
January 1995
-------
TABLE 5 (cont)
RECOVERY OF TNT SPIKED INTO REAL SOILS
SAMPLE ID
106-3
116-3
128-3
AMOUNT SPIKED
3.0
3.0
3.0
D TECH (ppm)
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
0.5 - 1.5
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
0.5 - 1.5
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
0.5 - 1.5
1.5 - 3.0
0.5 - 1.5
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
HPLC METHOD
8330
1.53
2.12
2.07
AGREEMENT3
Y, FN, FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
N
Y
FN
Y
Y
Y
Y
Y
Y
Y
Y
Y
4050-11
Revision 0
January 1995
-------
TABLE 6A
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
SAMPLE ID
61-1
61-10
61-11
61-12
61-13
61-14
61-15
61-16
61-17
61-18
61-19
61-2
61-20
61-21
61-22
61-23
61-24
61-25
61-26
61-27
61-28
61-29
61-3
61-30
61-4
61-5
61-6
61-7
61-8
61-9
TET-1
TET-2
TET-3
TL-1
D TECH RANGE
(ppm)
< 0.2
< 0.2
< 0.2
< 0.2
< 0.2
< 0.2
< 0.2
< 0.2
< 0.2
< 0.2
< 0.2
> 1.5
< 0.2
0.5-1.0
< 0.2
< 0.2
1.0-1.5
< 0.2
< 0.2
0.2-0.5
< 0.2
< 0.2
1.0-1.5
< 0.2
> 1.5
0.5 - 1.0
> 1.5
< 0.2
0.5-1.0
0.2-0.5
0.5-1.0
< 0.2
< 0.2
0.2-0.5
METHOD 8330
TNT (ppm)
< 0.09
< 0.09
< 0.09
< 0.09
< 0.09
< 0.09
< 0.09
< 0.09
< 0.09
< 0.09
< 0.09
> 3.0
< 0.09
2.44
< 0.09
< 0.09
1.4
< 0.09
< 0.09
0.27
< 0.09
< 0.09
1.3
< 0.09
1.1
1.0
> 3.0
< 0.09
1.0
0.56
< 0.09
< 0.09
< 0.09
0.99
AGREEMENT3
Y, FN, FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
FN
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
FP
Y
Y
Y
Y
Y
FP
Y
Y
FN
4050-12
Revision 0
January 1995
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SAMPLE ID
TL-2
TL-3
TL-4
TL-5
TL-6
TL-7
TL-8
TL-9
D TECH RANGE
(ppm)
r > 1.5
> 1.5
0.2-0.5
> 1.5
0.2-0.5
0.2-0.5
0.5-1.0
0.2-0.5
METHOD 8330
^JNT (ppm)
1.2
> 3.0
0.66
> 3.0
0.66
0.71
1.46
0.92
AGREEMENT3
Y, FN, FP
FP
Y
FN
Y
FN
FN
FN
FN
4050-13
Revision 0
January 1995
-------
TABLE 6B
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
Sample
Number
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
D TECH
Range (ppm)
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
0.5 - 1.0
0.5 - 1.0
0.5 - 1.0
> 1.5
0.5 - 1.0
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
8330 TNT
(ppm)
5.75
3.32
166
2500
2.72
<2.0
<2.0
140
230
1100
23.5
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
3.23
<2.0
<2.0
4.75
<2.0
<2.0
<2.0
3.64
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
6.39
8330 TNB
(ppm)
< 1.0
< 1.0
< 1.0
18.50
< 1.0
7.02
5.12
12.2
20.2
16.9
11.5
2.95
1.30
1.89
3.94
4.54
4.57
10.5
24.3
81
1.61
2.60
2.97
6.29
< 1.0
5.05
6.62
1.94
8.53
6.77
6.75
17.6
39.2
TNT Equivalent
(ppm)
5.75-6.0
3.32-3.57
166
2504
2.72-2.97
1.76-3.76
1.28-3.28
143
235
1104
26.0
0.74-2.74
0.33-2.33
0.47-2.47
0.99-2.99
1.14-3.14
1.14-3.14
2.63-4.63
9.3
20.3
0.40-2.40
5.40
0.74-2.74
1.57-3.57
<2.25
4.90
1.66-3.66
0.49-2.49
2.13-4.13
1.69-3.69
1.69-3.69
4.40-6.41
16.2
AGREEMENT3
Y, FN, FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
FN
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
4050-14
Revision 0
January 1995
-------
Sample
Number
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
D TECH
Range (ppm)
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
0.5 - 1.0
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
1.0 - 1.5
> 1.5
> 1.5
> 1.5
8330 TNT
(ppm)
4.20
5.14
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
820
1200
27.6
7.43
4.98
3.32
3.42
4.32
7.57
5.12
<2.0
<2.0
33.5
2.19
7.00
2.84
<2.0
2.23
5.38
2.60
4.43
4.79
2.29
8.84
9.01
29.00
<2.0
8330 TNB
(ppm)
1.39
< 1.0
2.68
7.65
27.70
9.01
30.90
35.70
5.69
24.0
11.9
9.01
9.46
10.4
16.5
28.2
44.8
81.2
1.64
2.27
23.4
8.43
11.0
4.69
5.67
12.8
31.4
13.0
31.1
25.9
18.2
148
< 1.0
6.02
1.30
TNT Equivalent
(ppm)
4.55
5.14-5.39
0.67-2.67
1.91-3.91
6.9-8.9
2.25-4.25
7.7-9.7
8.9-10.9
821
1206
31
9.70
7.40
5.90
7.60
11.4
18.8
25.4
0.41-2.41
0.57-2.57
39.4
4.30
9.75
4.01
1.42-3.42
5.43
13.23
5.85
12.2
11.3
6.8
45.8
9.01
30.50
0.33-2.33
AGREEMENT3
Y, FN, FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
FN
Y
Y
Y
4050-15
Revision 0
January 1995
-------
Sample
Number
78
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
95
96
97
98
99
100
101
D TECH
Range (ppm)
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
0.5 - 1.0
0.5 - 1.0
1.0 - 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
1.0 - 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
> 1.5
0.5 - 1.0
> 1.5
> 1.5
> 1.5
> 1.5
< 0.2
> 1.5
> 1.5
8330 TNT
(ppm)
<2.0
<2.0
2.49
<2.0
<2.0
<2.0
<2.0
3.98
5.67
7.05
8.04
1000
2.12
8.83
3.64
3.22
<2.0
<2.0
<2.0
<2.0
<2.0
351
116
4.29
<2.0
2.34
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
4.24
<2.0
8330 TNB
(ppm)
7.50
4.70
30.0
29.1
8.86
30.7
38.1
183
122
< 1.0
< 1.0
7.49
2.99
5.56
3.20
10.6
18.3
17.4
20.4
117
1.96
5.77
39.2
3.92
11.6
9.26
48.7
5.05
12.6
10.7
11.1
3.74
1.88
< 1.0
1.10
TNT Equivalent
(ppm)
1.88-3.88
1.18-3.18
9.99
7.28-9.28
2.22-4.22
7.68-9.68
9.59-11.6
49.7
36.2
7.05-7.3
8.04-8.29
1001
2.87
10.20
4.44
5.87
4.58-6.58
4.43-6.43
5.10-7.10
29.2-31.2
0.49-2.49
352
126
5.27
2.9-4.9
4.66
12.2-14.2
1.26-3.26
3.15-5.15
2.68-4.68
2.78-4.78
0.94-2.94
0.47-2.47
4.24-4.49
0.28-2.28
AGREEMENT3
Y, FN, FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
FN
FN
FN
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
FN
Y
Y
Y
Y
FN
Y
Y
4050-16
Revision 0
January 1995
-------
Sample
Number
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
through
279
280
through
365
366
through
381
382
through
391
D TECH
Range (ppm)
0.5 - 1.0
1.0 - 1.5
> 1.5
> 1.5
> 1.5
0.5 - 1.0
1.0 - 1.5
0.5 - 1.0
0.5 - 1.0
0.5 - 1.0
1.0 - 1.5
> 1.5
> 1.5
> 1.5
0.2 - 0.5
0.5 - 1.0
0.2 - 0.5
0.5 - 1.0
> 1.5
> 1.5
> 1.5
0.2 - 0.5
< 0.2
< 0.2
0.2 - 0.5
< 0.2
0.2 - 0.5
0.5 - 1.0
1.0 - 1.5
8330 TNT
(ppm)
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
6.35
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
<2.0
8330 TNB
(ppm)
1.28
2.70
10.5
14.1
18.4
6.35
6.66
21.8
5.29
4.49
16.3
28.7
17.7
24.1
< 1.0
2.40
4.70
11.6
56.9
45.6
67.7
2.78
1.61
4.07
3.12
<1.0
< 1.0
< 1.0
< 1.0
TNT Equivalent
(ppm)
0.32-2.32
0.68-2.68
2.63-4.63
3.53-5.53
4.6-6.6
1.59-3.59
1.67-3.67
5.45-7.45
1.32-3.32
1.12-3.12
4.08-6.08
7.18-9.18
4.43-6.43
6.03-8.03
6.35-6.6
0.60-2.6
1.18-3.18
2.9-4.9
14.2-16.2
11.4-13.4
16.9-18.9
0.7-2.7
0.4-2.4
1.02-3.02
0.78-2.78
<2.25
<2.25
<2.25
<2.25
AGREEMENT8
Y, FN, FP
Y
Y
Y
Y
Y
FN
FN
FN
FN
FN
FN
Y
Y
Y
FN
Y
FN
FN
Y
Y
Y
FN
FN
FN
FN
Y
Y
Y
Y
4050-17
Revision 0
January 1995
-------
Sample
Number
392
through
399
D TECH
Range (ppm)
> 1.5
8330 TNT
(ppm)
<2.0
8330 TNB
(ppm)
< 1.0
TNT Equivalent
(ppm)
<2.25
AGREEMENT3
Y, FN, FP
Y
4050-18
Revision 0
January 1995
-------
TABLE 6C
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
Third Party Field Trial
Sampl e
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
Oil ution
Factor
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
SDI
Results
<0.5
<0.5
<0.5
0.5-1.5
<0.5
0.5-1.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
0.5-1.5
<0.5
<0.5
0.5-1.5
<0.5
<0.5
<0.5
<0.5
0.5-1.5
<0.5
<0.5
8330
TNT Results
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
8330
TNT+TNB
Results
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
AGREEMENT3
Y, FN, FP
Y
Y
Y
FP
Y
FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
FP
Y
Y
FP
Y
Y
Y
Y
FP
Y
Y
4050-19
Revision 0
January 1995
-------
Sample
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
Dilution
Factor
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
10
10
1
SDI
Results
<0.5
0.5-1.5
<0.5
0.5-1.5
<0.5
0.5-1.5
0.5-1.5
0.5-1.5
<0.5
<0.5
3.0-4.0
<0.5
<0.5
<0.5
0.5-1.5
1.5-3.0
<0.5
0.5-1.5
0.5-1.5
0.5-1.5
0.5-1.5
<0.5
0.5-1.5
<0.5
0.5-1.5
5-15
40-50
0.5-1.5
8330
TNT Results
<0.15
<0.15
<0.15
0.15-0.99
<0.15
<0.15
<0.15
0.15-0.99
<0.15
<0.15
0.15-0.99
<0.15
<0.15
<0.15
<0.15
0.15-0.99
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
<0.15
1.4
35
<0.15
8330
TNT+TNB
Results
<0.25
<0.25
<0.25
0.15-0.99
<0.25
<0.25
<0.25
0.15-0.99
<0.25
<0.25
0.25-2.0
<0.25
<0.25
<0.25
<0.25
0.15-0.99
<0.25
0.15-0.99
<0.25
<0.25
1.3
<0.25
<0.25
<0.25
<0.25
3.2
41.67
<0.15
AGREEMENT3
Y, FN, FP
Y
FP
Y
Y
Y
FP
FP
Y
Y
Y
FP
Y
Y
Y
FP
FP
Y
Y
FP
FP
Y
Y
FP
Y
FP
Y
Y
FP
4050-20
Revision 0
January 1995
-------
Sample
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
Dilution
Factor
1
1
1
1
1
10
1
10
10
100
1000
10000
1000
10
100
10
10
10
10
10
10
10
10
100
10
10
10
10
SDI
Results
0.5-1.5
1.5
0.5-1.5
3.0-4.0
0.5-1.5
15-30
0.5-1.5
4-40
5-15
400-500
4000-5000
15000
15000
5-15
400-500
15-30
5-15
40
5-15
5
4-30
5-15
5-15
300-400
5-15
5-15
5-15
15-30
8330
TNT Results
0.15-0.99
0.15-0.99
<0.15
0.15-0.99
<0.15
22
-
2.1
2
360
6300
4000
530
2.8
460
4.2
1.0
5.1
1.9
1.6
2.2
1.7
2.2
180
3.1
2.8
2.5
3.2
8330
TNT+TNB
Results
0.15-0.99
0.15-0.99
<0.15
0.15-0.99
<0.15
22.48
<0.15
32
3.1
364
6327
4027
547
3.375
477
6.73
1.57
34.5
4
2.7
4.3
2
3.95
192.19
4.61
5.26
5.26
4.5
AGREEMENT3
Y, FN, FP
Y
Y
FP
FP
FP
Y
FP
Y
Y
Y
Y
FP
FP
Y
Y
FP
FP
Y
Y
Y
Y
FP
Y
Y
Y
Y
Y
FP
4050-21
Revision 0
January 1995
-------
Sample
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
Dilution
Factor
10
10
10
10
10
10
100
1
10000
10
10000
1000
1
10000
10000
10
10
100
SDI
Results
40-50
15-30
15-30
15-30
5-15
5-15
150-300
4-5
15000
40-50
15000-30000
500-1500
3.0
40000-50000
4000-5000
15-30
15-30
50-150
8330
TNT Results
1
3.8
36
3.6
2.6
3.2
78
18000
11000
36
11000
88
9.6
15000
2200
3.6
6.4
26
8330
TNT+TNB
Results
23
18.5
52.5
8.66
19.16
3.84
82
18050
11052.9
42.4
11052.9
107
10.17
15050
2220
3.9
6.7
28.76
AGREEMENT3
Y, FN, FP
Y
Y
FN
Y
Y
Y
Y
FN
Y
Y
Y
FP
FN
FP
Y
FP
FP
Y
4050-22
Revision 0
January 1995
-------
METHOD 4051
HEXAHYDRO-1.3.5-TRINITRO-1,3.5-TRIAZINE (RDX) IN SOIL & WATER BY IMMUNOASSAY
1.0 SCOPE AND APPLICATION
1.1 Method 4051 is a procedure for screening waters and soils to
determine when hexahydro-l,3,5-trinitro-l,3,5-triazine (RDX, CAS No. 121-82-4)
is present at concentrations above 5 p,g/L in water and 0.5 mg/kg in soil.
Method 4051 provides an estimate of the concentration of RDX by comparison with
a reference.
1.2 Using the test kit from which this method was developed, 96% of water
samples containing 2.5 ppb or less of RDX will produce a negative result and 99+%
of waters containing 10 ppb or more will produce a positive result. In addition
99+% of soil samples containing 0.25 ppm or less of RDX will produce a negative
result and 99+% of soil samples containing 1.0 ppm will produce a positive
result.
1.3 In cases where the exact concentration of RDX is required,
quantitative techniques (i.e., Method 8330) should be used.
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 Test kits are commercially available for this method. The
manufacturer's directions should be followed. In general, the method is
performed using a diluted water sample or an extract of a soil sample. Samples
and an enzyme conjugate reagent are added to immobilized RDX antibody. The
enzyme-RDX conjugate "competes" with RDX present in the sample for binding to
immobilized RDX antibody. The enzyme-RDX conjugate bound to the antibody then
catalyzes a colorless substrate to a colored product. The test is interpreted
by comparing the color produced by a sample to the response produced by a
reference reaction.
3.0 INTERFERENCES
3.1 Chemically similar compounds and compounds which might be expected
to be found in conjunction with RDX contamination were tested to determine the
concentration required to produce a positive test result.
3.1.1 Table 1 provides the concentrations of compounds tested with
the D TECH test kit that are required to elicit a positive response at the
4051-1
Revision 0
January 1995
-------
MDL, as well as the concentration required to yield 50% inhibition
compared to the standard curve.
4.0 APPARATUS AND MATERIALS
4.1 Immunoassay test kit: D TECH™ RDX (Strategic Diagnostics Inc.), or
equivalent. Each commercially available test kit will supply or specify the
apparatus and materials necessary for successful completion of the test.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HAULING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Water and soil samples may be contaminated, and should therefore be
considered hazardous and handled accordingly.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance indicated in Tables 3-6.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for the test kit being used
for quality control procedures specific to the test kit used. Additionally,
guidance provided in Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.3 Do not use test kits past their expiration date.
8.4 Do not use tubes or reagents designated for use with other test kits.
Do not mix reagents from one kit lot with a different kit lot.
8.5 Use the test kits within their specified storage temperature and
operating temperature limits.
4051-2 Revision 0
January 1995
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8.6 Method 4051 is intended for field or laboratory use. The appropriate
level of quality assurance should accompany the application of this method to
document data quality.
9.0 METHOD PERFORMANCE
9.1 Table 1 provides data on the minimum concentrations of possible
interferants and co-contaminants required to elicit a positive response in the
test kits evaluated.
9.2 Twenty six soil samples, known to not be contaminated with RDX, were
extracted and analyzed using the DTECH RDX kit to determine the extent of soil
matrix effects on the performance of the test kit. The results are provided in
Table 2A, and show that false positive results are not attributable to soil
components. Table 2B presents similar data generated from the analysis of thirty
uncontaminated water samples.
9.3 Thirty water samples and thirty soil samples, known to not be
contaminated with RDX, were each spiked with RDX at one-half and two times the
MDL (0.25 and 1.0 ppm respectively). These samples were analyzed with the DTECH
RDX test kit to determine the error rate of the assay. The results are presented
in Tables 3A and 3B.
9.4 Ten different soil types, all known to not be contaminated with RDX,
were spiked with RDX. The spiked soil samples were each analyzed six times with
the DTECH kit to determine the extraction efficiency of the method. The data are
presented in Table 4.
9.5 Table 5A presents the results of analysis of three soils spiked at
approximately 0.4, 1 and 3 ppm RDX. Each sample was analyzed using Method 8330
and in triplicate using the DTECH kit. Table 5B presents similar data generated
using water samples spiked at 10, 20 and 40 ppb of RDX.
9.6 Tables 6A through 6D present the results of four field trials.
Freshly collected (Table 6A, 6B and 6D) and archived (6C) soil samples, and
samples of water collected from monitoring wells (Table 6B), were analyzed by
commercial laboratories using Method 8330 and the DTech test kit. The Tables
provide results for both analyses, and evaluate the agreement between the two.
10.0 REFERENCES
1. D TECH™ TNT Users Guide , SDI/Em Sciences.
2. Haas, R.J., and B.P. Simmons, "Measurement of Trinitrotoluene (TNT) and
Hexahydro-l,3,5-trinitro-l,3,5-triazine (RDX) in Soil by Enzyme
Immunoassay and High Performance Liquid Chromatography (EPA Method 8330)",
California Environmental Protection Agency, Department of Toxic Substances
Control, Hazardous Materials Laboratory, March, 1995.
4051-3 Revision 0
January 1995
-------
TABLE 1
CROSS REACTANTS - D TECH™ RDX test kit
SAMPLE
RDX d
HMX d
TNT (trinitrotoluene)
Tetryl d
TNB (trinitrobenzene)
2-amino-4,6-dinitrotoluene
4-amino-2,6-dinitrotoluene
2,4-dinitrotoluene
2,6-dinitrotoluene
1,3-dinitrobenzene
nitrobenzene
2-nitrotoluene
3-nitrotoluene
4-nitrotoluene
nitroglycerine
pent aery thritoltetranitrate
MDLa
(Ppb)
5
150
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
ICgOb
(ppb)
25
800
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
> 500
% CROSS
REACTIVITY0
100
3
< 1
< 1
< 1
< 1
< 1
< 1
< 1
< 1
< 1
< 1
< 1
< 1
< 1
< 1
The following compounds were not detected at or above 500 ppm (lOOx the method
MDL for RDX):
Atrazine Benzo(a)pyrene Benzo(b)fl uoranthene Benzene
Aroclor 1254 Acenaphthene Dibenz(ah)anthracene Chrysene
Acetone Acenaphthalene Fluoranthene Fluorene
Toluene 1,2-Benzanthracene Benzo(k)fluoranthene Pyrene
Ethylbenzene Indeno(123-cd)pyrene Benzo(ghi )perylene Xylene
Naphthalene Methanol Phenanthrene
The Method Detection Limit (MDL) is defined as the lowest concentration of
compound that yields a positive test result.
The IC50 is defined as the concentration of compound required to produce a
test response equivalent to 50% of the maximum response.
% Cross Reactivity is determined by dividing the equivalent RDX
concentration by the actual compound concentration at IC50.
RDX = hexahydro-l,3,5-trinitro-l,3,5-triazine
HMX = octahydro-l,3,5,7-tetranitro-l,3,5,7-tetrazocine
Tetryl = methyl-2,4,6-trinitrophenylnitramine
4051-4
Revision 0
January 1995
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TABLE 2A
SOIL MATRIX EFFECTS
Soil ID #
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
Soil Type
Low OM Clay Loam
Sassafras Sandy Loam
Cecil Sandy Clay Loam
Davidson Clay Loam
Shontik-Casa Grande Clay
Trix Sandy Clay Loam
Trix-Casa Grande Clay Loam
Yolo Loam
Capay Silty Clay
Sycamore Silt Loam
Dennis Silt Loam
Luray Silty Clay Loam
Wooster Silt Loam
Vienna Loam
Opal Clay
Raulb Silt Loam
Rockfield Silt Loam
Cisne Silt Loam
Muscatine Silt Loam
Avonburg
Matapeake Silt Loam
Evesboro Low OM Sand
Selbyville High OM Sand
Casa Grande Clay Loam
Grundy Silty Clay Loam
Drummer Silty Clay
Non-Soil Control
State
DE
DE
GA
GA
AZ
AZ
AZ
CA
CA
CA
KA
OH
OH
SD
SD
IN
IN
IL
IL
DE
DE
DE
AZ
KA
IL
-
D TECH Result
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
4051-5
Revision 0
January 1995
-------
TABLE 2B
WATER MATRIX EFFECTS
Water ID
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
Location
Adamsville, RI
Buttermilk Falls, PA
Hudson River, PA
Germantown, PA
Houston, TX (1)
Houston, TX (2)
Ontario, CA
Pacific Ocean, CA
S. Darthmouth, MA
Newark, DE (1)
U.S. Army Waterways
U.S. Army WES
U.S. Army WES
U.S. Army WES
U.S. Army WES
U.S. Army WES
U.S. Army WES
U.S. Army WES
Georgetown, DE
Newark, DE (2)
Burlington, IA
Burlington, IA
Lake St. Germain, Canada
Milliston, WI
Moorhead, MN
McKenzie Co., ND
Wolcott, IN
Newark, DE (3)
Smith Island, MD
Adrian, GA
DI Control
D TECH Result (ppb)
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
< 5.0
4051-6
Revision 0
January 1995
-------
TABLE 3A
False Negative and False Positive Rates, Soil Matrix3
Spike Concentration
0.25 ppm
1.0 ppm
False Positive Rate
0%
-
False Negative Rate
-
0%
3 Thirty negative soils were spiked with RDX at one-half and two times the MDL
(0.25 and 1.0 ppm respectively). These samples were analyzed with the DTECH
RDX test kit to determine the error rate of the assay.
TABLE 3B
False Negative and False Positive Rates, Water Matrix3
Spike Concentration
0.25 ppm
1.0 ppm
False Positive Rate
3.3%
-
False Negative Rate
_
0%
a Thirty negative water samples were spiked with RDX at one-half and two times
the MDL (0.25 and 1.0 ppm respectively). These samples were analyzed in
triplicate with the DTECH RDX test kit to determine the error rate of the
assay.
4051-7
Revision 0
January 1995
-------
TABLE 4
DETERMINATION OF EXTRACTION EFFICIENCY FROM SOIL SAMPLES3
Soil ID : Spike
(ppm)
101:1
106:1
108:1
109:1
110:1
116:1
117:1
123:1
126:1
128:1
Non-Soil
Average
101:6
106:6
108:6
109:6
110:6
116:6
117:6
123:6
126:6
128:6
Non-Soil
Average
Mean RDX
Concentration
(ppm)
0.53
0.88
0.86
0.66
0.70
0.96
0.92
1.00
1.03
1.02
1.05
0.86
4.9?
6.15
5.69
6.11
6.12
6.26
5.71
6.05
6.82
6.02
6.02
5.98
Standard
Deviation
0.19
0.13
0.23
0.22
0.14
0.12
0.42
0.45
0.25
0.18
0.13
0.23
0.54
0.84
1.09
0.93
0.46
1.21
0.72
0.8
0.33
0.62
0.83
0.75
Coefficient
of
Variation
(%)
35
15
26
34
19
13
46
45
24
18
12
27
11
14
19
15
8
19
13
13
5
10
14
13
Recovery
(a/ \
\'°)
53
88
86
66
70
96
92
100
103
102
105
86
8?
103
95
102
102
104
95
101
114
100
100
100
4051-8
Revision 0
January 1995
-------
TABLE 5A
RECOVERY OF RDX SPIKED INTO REAL SOILS.
Soil ID
106
116
128
Spike
Concentration
(ppm)
0.4
1.0
3.0
0.4
1.0
3.0
0.4
1.0
3.0
Method 8330
(ppm)
0.32
0.83
1.79
0.29
0.66
0.61
0.31(0.25)
0.73(0.73)
0.75(2.27)
D TECH
(ppm)
< 0.5
< 0.5
< 0.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
> 2.0
> 2.0
> 9 n
< 0.5
< 0.5
< 0.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
> 2.0
< 2.0
> ? n
< 0.5
< 0.5
< 0.5
< 0.5
0.5 - 1.5
0.5 - 1.5
> 2.0
< 2.0
< ? n
AGREEMENT3
Y, FN, FP
Y
Y
Y
Y
Y
Y
FP
FP
FP
Y
Y
Y
Y
Y
Y
FP
FP
FP
Y
Y
Y
FN
Y
Y
Y
Y
Y
4051-9
Revision 0
January 1995
-------
TABLE 5B
RECOVERY OF RDX SPIKED INTO WATERS
Sample ID
1
7
Spike
Concentration
(ppb)
10
20
40
10
10
20
40
20
Method 8330
(ppb)
11.1
18.0
35.7
9.0
9.?
19.4
36.5
17.1
D TECH
(ppb)
5 - 15
5 - 15
5 - 15
15 - 30
15 - 30
15 - 30
> 45
> 45
> 45
5 - 15
5 - 15
5 - 15
5 - 15
5 - 15
5 - 15
15 - 30
15 - 30
15 - 30
> 45
> 45
> 45
15 - 30
15 - 30
15 - 30
AGREEMENT3
Y, FN, FP
Y
Y
Y
Y
Y
Y
Y
FP
FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
FP
FP
FP
Y
Y
Y
4051-10
Revision 0
January 1995
-------
TABLE 5B
RECOVERY OF RDX SPIKED INTO WATERS
Sample ID
3
Spike
Concentration
(ppb)
10
20
40
40
Method 8330
(ppb)
9.7
18.2
35.8
31.8
D TECH
(Ppb)
5 - 15
5 - 15
5 - 15
15 - 30
15 - 30
15 - 30
> 45
> 45
30 - 45
> 45
30 - 45
30 - 45
AGREEMENT3
Y, FN, FP
Y
Y
Y
Y
Y
Y
FP
FP
Y
FP
Y
Y
4051-11
Revision 0
January 1995
-------
TABLE 6A
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
Sample ID
S4
S12
S14
S15
S19
S20
S21
Tl-2
T2-4
T6-1
T3-5
T12-3
T12-6
T20-3
T21-10
T22-4
T22-5
T22-6
T28-3
T28-4
T28-5
T28-6
T28-7
T28-8
T28-9
Method 8330
(ppm)
< 0.2
< 0.2
1.72
< 0.2
2.12
1.61
0.32
0.21
1.41
2.62
2.00
< 0.2
1.00
< 0.2
1.89
< 0.2
0.83
0.99
3.73
< 0.2
< 0.2
< 0.2
< 0.2
< 0.2
< 0.2
D TECH
(ppm)
< 0.5
< 0.5
1.5 - 2.0
< 0.5
1.5 - 3.0
1.5 - 3.0
< 0.5
< 0.5
1.5 - 2.0
> 3.0
0.5 - 1.5
< 0.5
0.5 - 1.5
< 0.5
1.5 - 2.0
< 0.5
0.5 - 1.5
0.5 - 1.5
> 3.0
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
AGREEMENT3
Y, FN, FP
Y
Y
Y
Y
Y
Y
Y
Y
FP
FP
FN
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
4051-12
Revision 0
January 1995
-------
TABLE 6A
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
Sample ID
T28-10
T28-11
T28-12
T28-13
T31-4
T12-5
Method 8330
(ppm)
0.28
1.51
1.3
0.6
1.22
0.26
D TECH
(ppm)
< 0.5
1.5 - 3.0
1.5 - 3.0
0.5 - 1.5
1.5 - 2.0
< 0.5
AGREEMENT3
Y, FN, FP
Y
Y
FP
Y
FP
Y
4051-13
Revision 0
January 1995
-------
TABLE 6B
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
Sample
ID
1
3
13
15
16
23
24
25
26
31
33
34
35
37
38
43
44
47
48
58
59
64
67
68
75
84
85
87
94
96
97
Method 8330
(ppm)
4.00
19.0
1.30
1.80
3.40
0.48
0.68
0.68
0.75
0.13
0.74
0.48
1.30
5.50
0.55
1.30
40.0
2.30
0.36
0.79
0.80
2.20
10.9
3.40
3.90
17.6
70.3
101
1.60
0.20
5.40
Replicate 1
D TECH
(ppm)
> 3.0
> 6.0
0.5 - 1.5
1.5 - 3.0
> 3.0
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
1.5 - 3.0
> 6.0
0.5 - 1.5
1.5 - 3.0
> 6.0
> 3.0
0.5 - 1.5
0.5 - 1.5
1.5 - 3.0
1.5 - 3.0
> 6.0
1.5 - 3.0
> 3.0
> 6.0
> 6.0
> 6.0
1.5 - 3.0
< 0.5
> 3.0
AGREEMENT3
Y, FN, FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
FP
Y
Y
FP
FP
Y
FP
Y
FP
FP
Y
FP
Y
Y
FN
Y
Y
Y
Y
Y
Y
Y
Replicate 2
D TECH
(ppm)
> 3.0
> 6.0
0.5 - 1.5
1.5 - 3.0
> 3.0
0.5 - 1.5
< 0.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
0.5 - 1.5
1.5 - 3.0
> 3.0
0.5 - 1.5
1.5 - 3.0
> 6.0
> 3.0
< 0.5
0.5 - 1.5
1.5 - 3.0
1.5 - 3.0
> 6.0
1.5 - 3.0
> 3.0
> 6.0
> 6.0
> 6.0
1.5 - 3.0
< 0.5
> 3.0
AGREEMENT3
Y, FN, FP
Y
Y
Y
Y
Y
Y
FN
Y
Y
Y
Y
Y
Y
Y
Y
FP
Y
FP
Y
Y
FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
4051-14
Revision 0
January 1995
-------
TABLE 6B
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
Sample
ID
98
99
105
111
113
115
119
Method 8330
(ppm)
< 0.05
< 0.05
130
< 1.0
< 1.0
3.00
36.0
Replicate 1
D TECH
(ppm)
< 0.5
< 0.5
> 60
> 3.0
< 5.0
< 5.0
> 30
AGREEMENT3
Y, FN, FP
Y
Y
Y
FP
Y
Y
Y
Replicate 2
D TECH
(ppm)
< 0.5
0.5 - 1.5
> 60
< 5.0
< 0.5
< 0,5
15 - 30
AGREEMENT3
Y, FN, FP
Y
FP
Y
Y
FN
FN
FN
4051-15
Revision 0
January 1995
-------
TABLE 6C
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
Sample ID
1
2
3
4
5
6
7
8
9
10
19
20
11
12
13
14
15
16
17
18
21
22
23
24
25
METHOD 8330
(ppm)
17
34
48
160
650
41
360
840
69
85
17
19
4.3
1.9
4.9
27
1.2
1.0
0.82
0.78
0.67
0.94
< 0.4
< 0.4
< 0.4
D TECH
(ppm)
15 - 30
15 - 30
> 30
60 - 120
150 - 300
> 30
50 - 150
> 600
> 60
30 - 60
> 6.0
> 6.0
> 3.0
> 3.0
> 3.0
1.5 - 3.0
1.5 - 3.0
1.5 - 3.0
0.5 - 1.5
0.5 - 1.5
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
AGREEMENT3
Y, FN, FP
Y
FN
Y
FN
FN
Y
FN
Y
Y
FN
Y
Y
Y
FP
Y
Y
FP
FP
Y
Y
FN
FN
Y
Y
Y
4051-16
Revision 0
January 1995
-------
TABLE 6C
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
Sample ID
26
27
28
29
30
METHOD 8330
(ppm)
< 0.4
< 0.4
< 0.4
< 0.4
< 0.4
D TECH
(ppm)
< 0.5
< 0.5
< 0.5
< 0.5
< 0.5
AGREEMENT3
Y, FN, FP
Y
Y
Y
Y
Y
4051-17
Revision 0
January 1995
-------
TABLE 6D
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
Sample
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
SDI
Dilution
Factor
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
SDI
Results
<0.5
<0.5
<0.5
<0.5
0.5-1.5
<0.5
<0.5
0.5-1.5
<0.5
<0.5
0.5-1.5
0.5-1.5
<0.5
<0.5
<0.5'
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
0.5-1.5
<0.5
<0.5
8330
Results
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
AGREEMENT8
Y, FN, FP
Y
Y
Y
Y
FP
Y
Y
FP
Y
Y
FP
FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
FP
Y
Y
4051-18
Revision 0
January 1995
-------
TABLE 6D
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
Sample
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
SDI
Dilution
Factor
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
SDI
Results
0.5-1.5
<0.5
<0.5
1.5-3.5
<0.5
0.5-1.5
<0.5
0.5-1.5
0.5-1.5
<0.5
1.5-3.0
<0.5
0.5-1.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
1.5-3.0
<0.5
0.5-1.5
<0.5
0.5-1.5
<0.5
0.5-1.5
8330
Results
<0.17
<0.17
<0.17
0.17-0.99
<0.17
<0.17
<0.17
0.17-0.99
0.17-0.99
<0.17
1.2
<0.17
<0.17
<0.17
<0.17
3.8
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
0.17-0.99
AGREEMENT8
Y, FN, FP
FP
Y
Y
FP
Y
FP
Y
Y
Y
Y
FP
Y
FP
Y
Y
FN
Y
Y
Y
FP
Y
FP
Y
FP
Y
Y
4051-19
Revision 0
January 1995
-------
TABLE 6D
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
Sample
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
SDI
Dilution
Factor
100
1
1
1
1
1
1
1
1
100
10
1
1
1
100
1
100
1
1
100
10
10
10
1
1
10
SDI
Results
50-150
3.0-4.5
3.0-4.5
<0.5
0.5-1.5
<0.5
<0.5
1.5-3.0
0.5-1.5
150-300
15-30
1.5-3.0
1.5-3.0
3.0-4.5
50-150
0.5-1.5
50-150
0.5-1.5
1.5-3.0
150-300
45-60
>60
30-45
1.5-3.0
4.5-6.0
>60
8330
Results
100
<0.17
<0.17
<0.17
<0.17
<0.17
<0.17
1.1
<0.17
290
46
4.8
0.17-0.99
12
150
2.6
140
7.8
3.2
340
55
67
63
2.4
6.4
73
AGREEMENT8
Y, FN, FP
Y
FP
FP
Y
FP
Y
Y
FP
FP
Y
FN
FN
FP
FN
Y
FN
Y
FN
FN
FN
Y
Y
FN
Y
FP
Y
4051-20
Revision 0
January 1995
-------
TABLE 6D
COMPARISON OF DTECH SOIL RESULTS WITH METHOD 8330
Sample
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
SDI
Dilution
Factor
10
1
1
1
10
1
100
10
100
1
1
1
1
1
1
1
1
1
1
1
1
1
SDI
Results
15-30
0.5-1.5
3.0-4.5
1.5-3.0
>60
>6
50-150
30-45
50-150
0.5-1.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
8330
Results
14
2.1
2.4
2
94
23
150
34
150
1.2
0.17-0.99
<15
<15
<2
<15
<5
<0.17
<15
<5
<0.17
<0.17
<0.17
AGREEMENT8
Y, FN, FP
FP
FN
FP
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
4051-21
Revision 0
January 1995
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4.5 MISCELLANEOUS SCREENING METHODS
The following methods are included in this section:
Method 3810: Headspace
Method 3820: Hexadecane Extraction and Screening of Purgeable
Organics
Method 8275A: Semivolatile Organic Compounds (PAHs and PCBs) in
Soils/Sludges and Solid Wastes Using Thermal
Extraction/Gas Chromatography/Mass Spectrometry
(TE/GC/MS) for Screening
Method 8515: Colorimetric Screening Method for Trinitrotoluene
(TNT) in Soil
Method 9078: Screening Test Method for Polychlorinated
Biphenyls in Soil
Method 9079: Screening Test Method for Polychlorinated
Biphenyls in Transformer Oil
FOUR - 16 Revision 3
January 1995
-------
METHOD 8275A
SEMIVOLATILE ORGANIC COMPOUNDS (PAHs and PCBs)
IN SOILS/SLUDGES AND SOLID WASTES USING
THERMAL EXTRACTION/GAS CHROMAT06RAPHY/MASS SPECTROMETRY (TE/GC/MS)
1.0 SCOPE AND APPLICATION
1.1 Method 8275 is a thermal extraction capillary GC/MS procedure for the
rapid quantitative determination of targeted PCBs and PAHs in soils, sludges and
solid wastes. The following analytes can be determined by this method:
Compound
8275A - 1
CAS No.'
Acenaphthene
Acenaphthylene
Anthracene
Benz[a]anthracene
Benzo[a]pyrene
Benzo[b]fluoranthene
Benzo[g,h,i]perylene
Benzo[k]fluoranthene
4-Bromophenyl phenyl ether
1-Chloronaphthalene
Chrysene
Dibenzofuran
Dibenz[ a, h] anthracene
Dibenzothiophene
Fluoranthene
Fluorene
Hexachlorobenzene
Indeno(l,2,3-cd)pyrene
Naphthalene
Phenanthrene
Pyrene
1 ,2,4-Trichlorobenzene
2-Chlorobiphenyl
3,3'-Dichlorobiphenyl
2,2' ,5-Trichlorobiphenyl
2,3' ,5-Trichlorobiphenyl
2,4' ,5-Trichlorobiphenyl
2,2' ,5,5'-Tetrachlorobiphenyl
2,2',4,5'-Tetrachlorobiphenyl
2,2',3,5'-Tetrachlorobiphenyl
2,3' ,4,4'-Tetrach1orobiphenyl
2,2',4,5,5'-Pentachlorobiphenyl
2,3' ,4,4' ,5-Pentachlorobiphenyl
2,2' ,3,4,4',5'-Hexachlorobiphenyl
2,2',3,4',5,5',6-Heptachlorobiphenyl
83-32-9
208-96-8
120-12-7
56-55-3
50-32-8
205-99-2
191-24-2
207-08-9
101-55-3
90-13-1
218-01-9
132-64-9
53-70-3
132-65-0
206-44-0
86-73-7
118-74-1
193-39-5
91-20-3
85-01-8
129-00-0
120-82-1
2051-60-7
2050-67-1
37680-65-2
3844-81-4
16606-02-3
35693-99-3
41464-40-8
41464-39-5
32598-10-0
37680-73-2
31508-00-6
35065-28-2
52663-68-0
Revision 1
January 1995
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Compound CAS No.'
2,2',3,3/,4,4'-Hexachlorobiphenyl
2,2',3,4,4',5,5'-Heptachlorobiphenyl
2,2',3,3',4,4',5-Heptachlorobiphenyl
2,2',3,3',4,4',5,5'-Octachlorobiphenyl
2,2',3,3',4,4',5,5',6-Nonachlorobiphenyl
2,2',3,3',4,4',5,5',6,6'-Decachlorobiphenyl
a Chemical Abstract Services Registry Number.
38380-07-3
35065-29-3
35065-30-6
35694-08-7
40186-72-9
2051-24-3
1.2 The estimated quantitation limit (EQL) of Method 8275 for individual
PAH compounds is 1.0 mg/kg (dry weight) (0.2 mg/kg for individual PCB congeners)
for soil/sediment samples and 75 mg/kg for wet sludges/other solid wastes
(depending on water and solvent content). However, this can be lowered by
adjusting the range of the calibration curve or introducing larger sample sizes
if sample interferences are not a factor. Detection limits achievable during
method development ranged from 0.01 to 0.5 mg/kg for compounds in the target
analyte list in Section 1.1 (dry samples).
1.3 This method is restricted to use by or under the supervision of
analysts experienced in the operation of a gas chromatograph and mass
spectrometer and skilled in the interpretation of mass spectral data. Each
analyst must demonstrate the ability to maintain control and generate acceptable
results with this method.
2.0 SUMMARY OF METHOD
2.1 A portion of sample (0.003-0.250 g, depending on the expected
concentration) is weighed into a sample crucible.
2.2 The crucible is placed in a thermal extraction chamber and then heated
to 340°C where it is held for 3 minutes.
2.3 Thermally-extracted compounds are swept into a GC equipped with a
split/splitless injection port (split ratio set at -35:1 for a low concentration
sample or -400:1 for a high concentration sample) and then concentrated on the
head of GC column. Thermal desorption lasts 13 minutes.
2.4 The temperature program of the GC oven is adjusted to the specific
temperature conditions required to elute the target analytes. The target
analytes are swept into a mass spectrometer for qualitative and quantitative
determination.
8275A - 2 Revision 1
January 1995
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3.0 INTERFERENCES
3.1 Raw GC/MS data from all blanks, samples, calibration standards and
internal standards must be evaluated for interferences.
3.2 Whenever a heavily concentrated sample is encountered the GC column
can become over loaded and an ON-LINE bakeout (Section 7.2.1) followed by a
method blank is necessary.
3.3 A maintenance bakeout (Section 7.5) is performed whenever the ON-LINE
bakeout and subsequent blank analyses do not eliminate system contamination.
4.0 APPARATUS AND MATERIALS
4.1 Thermal extraction/gas chromatograph/mass spectrometer (TE/GC/MS)
system
4.1.1 Mass spectrometer - Capable of scanning from 35 to 500 amu
every 1 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 decafluorotriphenylphosphine (DFTPP) which
meets the criteria of Method 8270.
4.1.2 Data system - A computer interfaced to the mass spectrometer
should allow the continuous acquisition and storage on machine-readable
media of all mass spectra obtained throughout the duration of the
chromatographic program. The computer must have software that can search
the 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 a
reconstructed ion chromatogram (RIC). Software must also be available that
allows integrating the abundances of the RIC between specified time or
scan-number 1imits.
4.1.3 GC/MS interface - Any GC-to-MS interface that gives acceptable
calibration points in the concentration range of interest may be used.
4.1.4 Gas chromatograph - Must be equipped with a heated
split/splitless capillary injection port, column oven, cryogenic cooling
(optional). The oven temperature should be controllable from ambient to
450°C, and have programmable oven heating controls capable of rates of
rc/min to 70°C/min.
4.1.5 Recommended capillary column - A fused silica coated with (5%
phenyl)-methylpolysiloxane phase; 25-50 meter length x (0.25 to 0.32 mm)
I.D. with 0.1 to 1.0 micron film thickness (OV-5 or equivalent), depending
on analyte volatility and separation requirements.
4.1.6 Thermal extraction unit - The TE unit should be constructed
such that the sample and any compounds extracted are permitted to contact
only heated fused quartz surfaces during the extraction and transfer to the
GC injection port. It is also imperative that all zones in the sample
transfer path be kept at a minimum of 315°C. The unit must also have a
bakeout capability of at least 650°C in the thermal extraction chamber and
8275A - 3 Revision 1
January 1995
-------
450°C in the interface zone. It should also be noted that all components,
crucibles, spatulas and tools that come in contact with the sample be
constructed of fused quartz to permit total oxidation of any residues.
4.2 Fused quartz sample spatula.
4.3 Muffle furnace tray - for holding the crucibles while cleaning.
4.4 Stainless steel forceps for sample crucible handling.
4.5 Petri dishes - for sample crucibles; one for clean storage and one for
dirty storage.
4.6 Sample staging disk.
4.7 Porous fused quartz sample crucibles.
4.8 Porous fused quartz sample crucible lids.
4.9 Muffle furnace - for cleaning sample crucibles, capable of heating to
800°C.
4.10 Cooling rack/pad - high temperature, ceramic or quartz.
4.11 Analytical balance - minimum 2 gram capacity, 0.01 mg sensitivity.
4.12 Mortar and pestle.
4.13 100- and 60-mesh sieves.
4.14 Sample vials - glass, with Teflon®-lined caps.
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 Stock standard solutions (1000 mg/L) - Standard solutions can be
prepared from pure standard materials or purchased as certified solutions.
5.2.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure material. Dissolve the material in pesticide quality
methylene chloride or other suitable solvent (some PAHs may require initial
dissolution in small volumes of toluene or carbon disulfide) 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.
5.2.2 Transfer the stock standard solutions into bottles with
Teflon®-!ined screw-caps. Store at -10°C to -20°C or less and protect from
8275A - 4 Revision 1
January 1995
-------
light. Stock standard solutions should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards from them.
5.2.3 Stock standard solutions must be replaced after 1 year or
sooner if comparison with a quality control reference standard indicates
a problem.
5.3 Intermediate standard solutions - An intermediate standard solution
should be prepared containing all the target analytes for the calibration
standard solutions (separate solutions for PAHs and PCBs) or all the internal
standards for the internal standard solution. The recommended concentration is
100 mg/L.
5.4 GC/MS tuning standard - A methylene chloride solution containing 50
mg/L of decafluorotriphenylphosphine (DFTPP) should be prepared. Store at -10°C
to -20°C or less when not being used.
5.5 Matrix spike standard - Prepare a spiking solution in methanol that
contains five or more of the target compounds at 100 mg/L for solid samples. The
selection of compounds should represent the boiling point range of the target
compounds. The stock and intermediate standards may be prepared as in Section
5.2 and 5.3 or commercially prepared certified standards are also acceptable.
The standards must be prepared independently from the calibration stock
standards.
5.6 Blank soil used for the preparation of the calibration standard soil
and internal standard soil is prepared as outlined below.
5.6.1 Obtain a clean (free of target analytes and interferences)
sedimentary soil. Dry and then grind it in a mortar and pestle. Sieve the
ground material through a 100 mesh sieve. Several 50 mg aliquots should
be extracted by TE/GC/MS (or other techniques) to determine if any
compounds are present that could interfere with the compounds in Tables 1
and 2.
5.6.2 If no interferences are found, 300-500 grams of dried and
sieved blank soil is tumbled for 2 days in a clean glass container with a
Teflon®-!ined cap to ensure homogeneity before the analytes are spiked onto
the soil.
5.7 Internal standard soil - The internal standard is prepared on a blank
soil (Section 5.6). The internal standard soil should contain all compounds
listed in Table 3 at a concentration of 50 mg/kg for each compound.
Commercially-prepared soil standards may be used if they are certified by the
manufacturer or by an independent source.
5.8 Calibration standard soil - The calibration standard is prepared on
a blank soil (Section 5.6). The calibration standard soil must contain all
target analytes to be reported, at a concentration of 35 mg/kg for the PAHs and
10 mg/kg for the PCBs. The PCBs are prepared at a lower concentration because
expected concentrations in soil are expected to be lower. If preferred, both the
PAHs and PCBs may be prepared at the same concentration. See Table 1 (PAH) and
Table 2 (PCB) for the analytes that have been tested by this method.
8275A - 5 Revision 1
January 1995
-------
Commercially-prepared soil standards may be used if they are certified by the
manufacturer or by an independent source.
5.9 Preparation of the internal standard and calibration standards on a
blank soil.
5.9.1 The 50 mg/kg internal standard soil and both the PAH
calibration standard (35 mg/kg) and PCB Calibration Standard (10 mg/kg)
soils are all prepared by the same technique. The intermediate standard
solutions (Section 5.3) or commercially-prepared certified solutions are
used for dosing a weighed amount of blank soil (Section 5.6). Weigh 20.0
g of blank soil (as prepared in Section 5.6) into a 4-oz. glass container.
Water is added (5% by weight) to aid in the mixing and dispersal of
analytes to the more polar sites in the soil, as occurs in nature. For an
intermediate standard containing 100 mg/L of each compound: add 10.0 ml
to the wetted blank soil for the internal standard soil; add 7.0 ml for the
PAH calibration standard soil; and 2 ml for the PCB calibration standard
soil. Add additional methylene chloride so that the total solvent provides
a slight solvent layer above the soil. This helps to distribute the
standard compounds homogeneously throughout the soil.
5.9.2 The solvent and water are allowed to evaporate at room
temperature until the soil appears dry (usually overnight). The soil
containers are tightly capped with Teflon®-!ined caps and placed on a
tumbler that slowly rotates and mixes the contents. All soils are tumbled
for at least five days to ensure homogeneity.
5.9.3 Internal standard soil and calibration standard soil should
be stored in amber glass vials with Teflon® seal caps at -10°C to -20°C or
less and protected from exposure to light and moisture. The soil standards
should be stable for up to 90 days under these storage conditions.
Internal standard and calibration standard soils should be checked
frequently against the calibration solutions for signs of degradation. The
check is performed by adding an equivalent concentration of standard
solution to the frit in the sample crucible lid just prior to transfer of
the crucible and lid to the thermal extraction unit.
5.9.4 Internal standard and calibration standard soils must be
replaced if the above check indicates degradation.
NOTE: The more volatile PAHs and PCBs in the soil calibration standards may
exhibit higher concentrations than the calibration solutions. This
results from the possibility of evaporation losses from the crucible
frit lid of the more volatile analytes.
5.10 Methylene chloride, methanol, carbon disulfide, toluene, and other
appropriate solvents - Pesticide quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION AND HANDLING
See the introductory material to this Chapter, Organic Analytes, Sec. 4.1.
8275A - 6 Revision 1
January 1995
-------
7.0 PROCEDURE
7.1 Sample crucible preparation
WARNING: Do not touch the crucibles with your fingers. This can result in a
serious burn during removal from the muffle furnace. Clean crucibles
can be contaminated with oils from the fingers. Always handle the
sample crucibles and lids with stainless steel tweezers.
7.1.1 Turn on the muffle furnace for cleaning crucibles and let it
heat to 800°C for at least 30 minutes.
7.1.2 To clean the crucibles, load the sample crucibles and lids
into the muffle furnace tray and place in the oven. Leave in the muffle
furnace for 15 minutes then remove tray and place on cooling pad (at least
15-20 minutes) before transferring crucibles to the "clean" petri dish.
7.1.3 All sample crucibles and lids should be pre-cleaned and placed
in a covered petri dish. Prepare a sufficient number of crucibles and lids
to prepare a 5-point calibration curve and/or for the number of sample
analyses planned.
7.2 Initial calibration of the TE/GC/MS system
7.2.1 Set the TE/GC/MS to the following recommended conditions and
bake out the system.
ON-LINE bakeout procedure -This procedure should be performed before each
set of calibration runs. If the autosampler is used, this should be incorporated
into the autosampling sequence.
IMPORTANT: Sample crucible must be removed from the thermal extraction unit
BEFORE bakeout procedure begins. It is not necessary to acquire
MS data during a bakeout although GC/MS data should be taken during
analysis of a method blank (following a bakeout) to monitor system
contamination.
GC initial column temp, and hold time: 35°C for 4 minutes
GC column temperature program:
GC final column temperature hold:
GC cool time:
GC injection port temperature:
MS transfer line temperature:
GC Carrier gas:
TE transfer line temperature:
TE interface oven temperature:
TE helium sweep gas flow rate:
TE sample chamber heating profile:
35 to 325°C at 20°C/min
325°C for 10 minutes
325°C to 35°C in 4 minutes
335°C; splitless mode for
entire run
290 - 300°C
Helium at 30 cm/sec
310°C
335°C
40 mL/min
Hold 60°C for 2 min; 60 -
650°C in 12 min; hold 650°C for
2 min; cool to 60°C.
8275A - 7
Revision 1
January 1995
-------
7.2.2 Set the TE/GC/MS system to the following recommended
conditions for calibration and sample analysis assuming a 30-m capillary
column (see Section 4.1.5).
Mass range:
MS scan time:
GC initial column temp, and hold
GC column temperature program:
GC final column temperature hold:
GC column cool rate:
GC injector type:
GC
GC
injection port temperature:
injection port setting:
MS transfer line temperature:
MS source temperature:
MS solvent delay time:
MS data acquisition:
Calibration Standard Soil weight:
Carrier gas:
TE transfer line temperature:
TE interface Oven temperature:
TE helium sweep gas flow rate:
TE sample heating profile:
45 - 450 amu
1.0 to 1.4 scan/sec
time: 35°C for 12 minutes
35 - 315°C at 8°C/min
315°C for 2 min (or until
benzo(g,h,i)perylene elutes.
315°C to 35°C in 4 minutes
Split/splitless capillary; 35:1
split ratio
325°C
Splitless for 30 sec, then
split mode for remainder of run
290 - 300°C
According to manufacturer's
specifications
15 minutes
Off at 49 minutes
See Section 7.2.5.3 for initial
calibration.
Helium at 30 cm/sec
310°C
335°C
40 mL/min
hold 60°C isothermal for 2
min; 60 - 340°C in 8 min; hold
340°C for 3 min; cool 340 -
60°C for 4 min.
NOTE: All calibration standards and samples must be analyzed under the same
split ratio settings.
7.2.3 Method blank - A blank should follow the ON-LINE bakeout using
the conditions listed in Section 7.4.2. Acquire the MS data and determine
that the system is free of target analytes and interferences at the project
required Method Detection Limit (MDL). Make appropriate corrections if
contamination is observed (i.e., bake out, change GC column, change TE
sample chamber and/or transfer line).
7.2.4 The GC/MS system must be hardware tuned to meet the DFTPP
criteria in Method 8270. Add 350 ng (because of the 35:1 split) of DFTPP
to the frit in the lid of the crucible and analyze following the conditions
in Section 7.2.2.
7.2.5 Initial calibration curve - A minimum of five calibration
standards should be run during the initial calibration of the system and
after any maintenance procedures which may affect system performance. This
calibration procedure should also be performed if there is more than a 20%
drift from the initial calibration curve and the calibration verification
unless system maintenance corrects the problem. Adjust the injection port
split ratio to 35:1 for the following calibration standard soil
8275A - 8
Revision 1
January 1995
-------
concentration. Any future modifications of the split ratio require the
preparation of a new initial calibration curve at the new split ratio.
7.2.5.1 Using forceps, remove a sample crucible from the
clean dish and place on the analytical balance. Tare or establish the
weight to the nearest 0.1 mg and place on a clean surface.
7.2.5.2 Weigh 10 mg (±3%) of internal standard soil (Section
5.7) into the sample crucible using a fused quartz sample spatula.
Place crucible back on the balance and determine weight. Record
current weight and tare balance for the next step.
7.2.5.3 Weigh the calibration standard soil into the
crucible (according to guidance below on PAHs and PCBs) and record
weight. Place a lid on the crucible and load into the Thermal
Extraction Unit or position in the autosampler. All analysis
information and conditions should be recorded in a sample log.
NOTE: If commercially-prepared standards are used, the weights may vary
slightly from what are presented below. This is acceptable as long as
the calibration curve is within the linear range of the GC/MS system.
PAH Standard:
50 mg (±3%) of 35 tug/kg PAH calibration standard soil (Section 5.9).
Repeat the process with 40, 20, 10, and 5 mg (±3%) of 35 mg/kg PAH
calibration standard soil + 10 mg of 50 mg/kg IS soil into separate
crucibles.
This results in 50, 40, 20, 10, and 5 ng respectively on column of
each target analyte in the calibration standard.
PCB Standard:
50 mg (+3%) of 10 mg/kg PCB calibration standard soil (Section 5.9).
Repeat the process with 40, 20, 10, and 5 mg (±3%) of 10 mg/kg PCB
calibration standard soil + 10 mg of 50 mg/kg IS soil into separate
crucibles.
This results in 10, 8, 4, 2, and 1 ng respectively on column of each
target analyte in the calibration standard.
NOTE: The sensitivity of the GC/MS system may require adjustment of the above
standard weights (calibration and internal) either up or down.
7.2.5.4 A split ratio of 300 or 400:1 is recommended for
high concentration samples. A new calibration curve at the higher
split ratio is required using a calibration standard soil containing
an appropriate concentration of target analytes (approximately 10
times more concentrated to achieve a similar concentration on column).
8275A - 9 Revision 1
January 1995
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7.2.6 Analysis - Upon method start, the sample is loaded into the
fused quartz sample chamber. The sample chamber is heated to 340°C and
held isothermal for 3 minutes. Helium carrier/sweep gas passes through the
sample chamber at a flow rate of 40 mL/min. Thermally-extracted compounds
are swept through a deactivated fused silica line into the GC injection
port where they are split (low -35:1) or (high -400:1) before being
concentrated on the head of the GC column which is held isothermal at 35°C.
Once thermal extraction is complete (13 min.), the sample chamber is
cooled, the GC oven is heated to 315°C at a rate of 10°C/min. (or according
to required separation needs). Exact thermal extraction method parameters
may be adjusted according to separation requirements.
7.2.7 Calculate response factors (RFs) for each analyte (using the
internal standard assignments given in Tables 4 and 5) and evaluate the
linearity of the calibration as described in Sec. 7.0 of Method 8000.
7.3 Calibration verification of the TE/GC/MS system
7.3.1 Prior to analysis of samples, the DFTPP tuning standard must
be analyzed. Follow the guidance in Section 7.2.4, The DFTPP criteria
must be demonstrated during each 12-hour shift.
7.3.2 At the beginning of each 12-hour shift, a method blank is
analyzed using the conditions in Section 7.2.2. Also, the mg of
calibration standard soil used for the midpoint of the initial calibration
curve and 10 mg of internal standard soil are analyzed and the RF values
are calculated for each target analyte. Calculate the % difference for
each target analyte as described in Sec. 7.0 of Method 8000. If the RF
values of each target analyte are not within 20% of their mean RF values
determined during the initial calibration, then the initial calibration
sequence must be repeated unless a calibration verification standard
analyzed after system maintenance meets the % difference criteria.
7.3.3 After every 6 hours of operation, a method blank is analyzed
to verify that the system is still clean.
7.4 Sample preparation, weighing and loading
7.4.1 Sample preparation - Decant and discard any water layer on a
sediment sample. Discard any foreign objects such as pieces of wood,
glass, leaves and rocks. Sample preparation requires homogenizing the wet
or dry sample as well as possible and selecting a representative aliquot
for analysis. Extremely wet samples (high H20 and solvents) can cause
excessive pressure in the MS if too much sample is inserted in the system.
See Sections 7.4.3.1 and 7.4.3.2 as guidelines for sample weight and
moisture considerations.
7.4.2 Determination of sample % dry weight - In certain cases
involving soil/sediment samples, sample results are desired based on a
dry-weight basis. When such data are desired, a portion of sample for this
determination should be weighed out at the same time as the portion used
for analytical determination. Also, for any sample that appears to contain
moisture, the % moisture must be calculated to determine whether drying of
8275A - 10 Revision 1
January 1995
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the sample is necessary prior to grinding in a mortar and pestle (see
Sections 7.4.3 and 7.4.4).
WARNING: The drying oven should be contained in a hood or vented. Significant
laboratory contamination may result from a heavily contaminated
hazardous waste sample.,
Weigh 5-10 g of a portion of sample into a tared crucible. Determine
the % dry weight of the sample by drying overnight at 105°C. Allow to cool
in a desiccator before weighing. Discard this portion after weighing as
a separate (unheated) portion will always be used for analysis. Calculate
the % dry weight as follows:
... ... g of dry sample ,nn
% dry weight = —xlOO
g of sample
% moisutre = 100 - (% dry weight)
7.4.3 Wet Samples (greater than 20% moisture)
7.4.3.1 For samples where naphthalenes are target analytes:
Perform the following steps quickly to minimize sample
exposure to air, thereby causing possible loss of naphthalenes as well
as sample weight variability because of loss of moisture. Tare the
crucible, weigh 10 mg of IS soil, then add 10-20 mg of a wet,
representative sample portion. Record the sample weight and insert
the crucible into the TE Inlet system.
7.4.3.2 For wet samples where naphthalenes are not target
analytes:
A representative aliquot (3-5 grams) of sample should be
spread in a thin layer in a clean shallow container and air dried at
room temperature (25°C) in a hood for 30 - 40 minutes.
7.4.3.2.1 Thick layers of clay type sediment may
require longer drying periods.
NOTE: No heat should be used to aid drying.
7.4.3.2.2 When dry, scrape the sample loose from
the container walls and break into uniform particle size or
grind in a mortar and pestle until reasonably uniform and
homogeneous in texture. Sieve through a 60-mesh sieve and
store in a sample vial.
8275A - 11 Revision 1
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7.4.4 Dry samples (less than 20 % water)
To prepare dry samples, homogenize 5-10 grams in a mortar and
pestle and sieve through a 60-mesh screen and store in a sample vial.
7.4.5 Internal standard weighing
7.4.5.1 Using forceps, remove a sample crucible from the
clean dish and place on the analytical balance. Tare or establish the
weight to the nearest 0.1 mg and place on a clean surface.
7.4.5.2 Weigh 10 mg (±3%) of internal standard soil mixture
into the sample crucible using a fused quartz sample spatula. Place
crucible back on the balance and determine weight. Record current
weight and tare balance for the next step.
7.4.6 Sample weighing - An aliquot (3 - 250 mg) of the prepared
sample is removed with a clean fused quartz spatula and placed in the
sample crucible and its weight determined. The weight of the sample to be
loaded into the thermal extraction crucible should be determined as
follows:
7.4.6.1 If low levels (0.02 - 5.0 mg/kg and low total
organic content) are expected, 100 to 250 mg of (dry) sample should
be weighed (assuming a 35:1 split ratio).
NOTE: As per Section 1.2, the estimated quantitation limit of this method is 1
mg/kg. Any concentrations that are determined to be lower than 1 mg/kg
would be considered estimated concentrations.
7.4.6.2 If high levels (500-1500 mg/kg and high total
organic content) are expected, 3 to 5 mg of (dry) sample should be
weighed (assuming a 35:1 split ratio).
7.4.6.3 For intermediate levels, adjust the weights
accordingly.
7.4.6.4 If the expected concentration exceeds 1500 mg/kg, a
greater split ratio is required. A split ratio of 300 to 400 is
recommended. This, of course, requires an initial calibration curve
developed with the selected split ratio.
7.4.6.5 For samples of unknown concentration or total
organic content, weigh less than 20 mg of sample for the initial run.
NOTE: It is highly recommended that samples of unknown concentration be screened
prior to TE/GC/MS analysis. This will prevent the need to reanalyze
samples as well as protect the system from overload which causes downtime
while performing system maintenance. The screening may be performed using
the optional FID device that is available as an add-on to the TE/GC/MS
device or by using a rapid semiquantitative extraction with methylene
chloride and injection on a GC/FID to determine relative concentrations.
8275A - 12 Revision 1
January 1995
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7.4.6.6 Select a sample for matrix spike determination (see
Method 3500 for guidance). Weigh one or two portions into crucibles
containing internal standard (see same Section cited above for
guidance on whether to analyze a matrix spike duplicate or a duplicate
sample). Add 5.0 ^L of the matrix spike standard (Section 5.5)
directly to the sample, immediately cover with lid and transfer to the
thermal extraction unit or the autosampler.
7.4.7 Loading sample - Make sample concentration assessment and
weigh sample into crucible containing the previously weighed internal
standard soil. Record sample weight (to the nearest 0.1 mg), cover the
crucible with lid and place covered crucible into the thermal extraction
unit or autosampler tray. If the sample is wet and/or target compounds
have a higher volatility than n-dodecane, the autosampler tray should be
chilled to 10-15'C.
7.4.8 Analysis - The sample is loaded into the fused quartz sample
chamber of the thermal extraction unit. See Section 7.2.7 for details on
the operation of the TE/GC/MS system.
7.4.8.1
signal to noise
appropriate from
For extremely low concentration samples where the
ratio is less than 3:1, increase sample size as
detector response after repeating Section 7.4.5.
7.4.8.2 If too much sample is extracted and GC column
overloading is evident, bake out system (as in Section 7.2.1) and
analyze a blank to determine if additional system cleaning is
necessary (Section 7.2.3). Use a smaller aliquot of the sample
(decreasing sample size as required) after repeating Section 7.4.5.
7.5 Maintenance bakeout procedure
7.5.1 System bakeout conditions: For OFF-LINE (no autosampling)
conditions following an extremely overloaded system and for routine
cleaning maintenance.
IMPORTANT: Sample crucible must be removed from the thermal extraction unit
BEFORE bakeout procedure begins.
Before this bakeout procedure, the TE interface oven should be cooled
so that the fused silica transfer line capillary can be removed. Following
the bakeout a new transfer line capillary should be installed.
GC initial column temp, and hold:
GC injection port temperature:
MS transfer line temperature:
GC Carrier gas:
TE transfer line temperature:
TE interface oven temperature:
TE sweep gas flow rate:
TE sample chamber heating profile:
8275A - 13
335°C, hold for 20 minutes
335°C; set in split mode
295 - 305°C
Helium at 30 cm/sec
OFF; until new capillary
installed
400°C
MAX; approx 60 mL/min;
Heat to 750'C and hold 700°C
for 3 min; cool to 60°C.
Revision 1
January 1995
-------
7.6 Qualitative analysis
Follow the procedures in Method 8270, Sec. 7.0, to identify target
compounds.
7.7 Quantitative analysis
Identified compounds are quantitated via the internal standard calibration
technique using the integrated abundance from the EICP of the primary
characteristic ion. The internal standard used should be assigned according to
Table 4. Calculate the concentration of each identified analyte as follows:
(A )(C. )(W. )
f- _ XX/VIS'VIS'
where: (RF)(A.J(WX)(D)
Cx = Concentration of compound being measured (mg/kg).
Ax = Area of characteristic ion for compound being measured in sample.
Cis = Concentration of internal standard soil (mg/kg).
WIS = Weight of internal standard soil (kg).
Wx = Weight of sample (kg).
RF = Mean response factor for compound being measured from initial
calibration curve.
Ais = Area of characteristic ion for the internal standard.
D = (100 - % moisture in sample)/100, or 1 for wet-weight basis.
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.
Required instrument QC is found in the following sections of Method 8275:
8.2.1 The GC/MS system must be tuned to meet the DFTPP
specifications in Sections 7.2.4 and 7.3.1.
8.2.2 There must be an initial calibration of the GC/MS system as
specified in Section 7.2.
8.2.3 The GC/MS system must meet the calibration verification
criteria specified in Section 7.3 each 12 hours.
8275A - 14 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. NIST (National Institute of Standards and
Technology) Standard Reference Material (SRM) #1939 may be used to monitor method
performance and document data quality. An SRM with PAHs may be substituted if
PAHs are the primary target analytes.
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
samples. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
8275A - 15 Revision 1
January 1995
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9.0 METHOD PERFORMANCE
9.1 Multilaboratory precision data for PAHs and for a few semivolatile
compounds are presented in Table 5. The results are based on the analysis of
test soils spiked at 10 mg/kg and analyzed by 3 different laboratories. A Ruska
ThermEx inlet interfaced to a GC/MS system was utilized to develop the data. A
total ion chromatogram generated by TE/GC/MS of PAH analysis is shown in Figure
1.
9.2 Multilaboratory performance data for PCB congeners are presented in
Table 6. The results are based on analyses of NIST Standard Reference Material
(SRM) #1939 using Method 8275A (Reference 1). A Ruska ThermEx inlet interfaced
to a GC/MS system was utilized to develop the data. An ion chromatogram
generated by TE/GC/MS of PCB congeners is shown in Figure 2.
10.0 REFERENCES
1. Worden, R., "Method 8275A: Quantitative Addendum For SW-846 Method 8275",
Research report to the U.S. Environmental Protection Agency; Ruska
Laboratories, Inc., Houston, TX, 1993.
2. Snelling, R., King, D., Belair, B., "Analysis of PAHs in Soils and Sludges
Using Thermal Extraction-GC-MS", Application Note 228-228; Hewlett-Packard
Co., Wilmington, DE, 1993.
3. King, D., Belair, B., "Analysis of PCBs in Soils and Sludges Using Thermal
Extraction-GC-MS", Application Note 228-229; Hewlett-Packard Co.,
Wilmington, DE, 1993.
8275A - 16 Revision 1
January 1995
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TABLE 1
PAH/SEMIVOLATILE CALIBRATION STANDARD SOIL AND QUANTITATION IONS
Compound Quantitation Ion
1,2,4-Trichlorobenzene1 180
Naphthalene 128
Acenaphthylene 152
Acenaphthene 153
Dibenzofuran 168
Fluorene 166
4-Bromophenyl phenyl ether1 248
Hexachlorobenzene1 284
Phenanthrene 178
Anthracene 178
Fluoranthene 202
Pyrene 202
Benzo[a]anthracene 228
Chrysene 228
Benzo[b]fluoranthene 252
Benzo[k]fluoranthene 252
Benzo[a]pyrene 252
Indeno(l,2,3-cd)pyrene 276
Dibenz[a,h]anthracene 278
Benzo[g,h,i]perylene 276
1 This analyte may be deleted if the target analytes are PAHs only.
All compounds are present at 35 mg/kg.
8275A - 17 Revision 1
January 1995
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TABLE 2
PCB CALIBRATION STANDARD SOIL
IUPAC # CAS #
1
11
18
26
31
52
49
44
66
101
118
138
187
128
180
170
194
206
209
2051-60-7
2050-67-1
37680-65-2
3844-81-4
16606-02-3
35693-99-3
41464-40-8
41464-39-5
32598-10-0
37680-73-2
31508-00-6
35065-28-2
52663-68-0
38380-07-3
35065-29-3
35065-30-6
35694-08-7
40186-72-9
2051-24-3
2
3
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
Compound Name Quantitation Ion
-Chlorobiphenyl
,3'
,2'
,3'
,4'
,2'
,2'
,2'
,3'
,2'
,3'
,2'
,2'
,2'
,2'
,2'
,2'
,2'
,2'
-Dichlorobiphenyl
, 5 -Tri chlorobiphenyl
, 5-Tri chl orobi phenyl
,5-Trichlorobiphenyl
,5,5' -Tetrachl orobi phenyl
,4, 5 '-Tetrachl orobi phenyl
,3, 5 '-Tetrachl orobi phenyl
,4,4' -Tetrachl orobi phenyl
, 4, 5, 5' -Pent achl orobi phenyl
,4,4' ,5-Pentachlorobiphenyl
,3,4,4' ,5'-Hexachlorobiphenyl
,3,4' ,5,5' ,6-Heptachlorobiphenyl
,3,3' ,4, 4 '-Hexachl orobi phenyl
,3,4,4' , 5, 5 '-Heptachl orobi phenyl
,3,3' ,4,4' ,5-Heptachlorobiphenyl
,3,3',4,4',5,5'-Octachlorobiphenyl
,3,3' ,4,4' ,5, 5 ',6-Nonachl orobi phenyl
,3,3',4,4',5,5',6,6'-Decachlorobiphenyl
188
222
258
258
258
292
292
292
292
326
326
360
394
360
394
394
430
392
426
All compounds are present at 10.0 mg/kg
8275A - 18
Revision 1
January 1995
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TABLE 3
INTERNAL STANDARD SOIL
Compound Quantitation Ion
2-Fluorobiphenyl 172
Phenanthrene-din1 188
Mo
Benzo[g,h,i]perylene ( C12) 288
1 This internal standard is more susceptible to soil microbial degradation. It
is suggested that a 13C-labeled phenanthrene be substituted.
TABLE 4
INTERNAL STANDARDS WITH CORRESPONDING PAH ANALYTES
ASSIGNED FOR QUANTITATION
2-F1uorobipheny1
Naphthalene
Acenaphthylene
Acenaphthene
Fluorene
All PCB Congeners from Table 2
Phenanthrene-dlO
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benzo(g,h,i)perylene( C12)
Benzo[a]anthracene
Chrysene
Benzo[b]fluoranthene
Benzo[k]fluoranthene
Benzo[a]pyrene
Indeno(l,2,3-cd)pyrene
Oibenz[a,h]anthracene
Benzo[g,h,i]perylene
8275A - 19 Revision 1
January 1995
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TABLE 5
MULTI- LABORATORY PRECISION DATA FROM OF ANALYSIS OF SAMPLES
CONTAINING PAHs/SEMIVOLATILES USING TE/GC/MS3
#
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
Compound
l,2,4-Trichlorobenzeneb
Naphthalene
Acenaphthylene
Acenaphthene
Dibenzofuran
Fluorene
4-Bromophenyl phenyl etherb
Hexachlorobenzeneb
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benzo[a]anthracene
Chrysene
Benzo[b]fluoranthene
Benzo[k]fluoranthene
Benzo[a]pyrene
Indeno(l,2,3-cd)pyrene
Dibenz [a, h] anthracene
Benzo[g,h,i]perylene
Quant. Ion
180
128
152
153
168
166
248
284
178
178
202
202
228
228
252
252
252
276
278
276
Results for 10 mg/kg
Mean
11.0
13.0
10.4
11.1
10.8
11.0
11.1
10.8
10.6
10.9
10.7
10.8
10.1
10.4
10.9
10.4
10.9
11.0
10.5
10.8
RSD (%)
35.7
45.3
18.1
22.6
22.9
24.0
17.6
18.5
20.2
34.4
17.0
19.9
24.1
17.8
23.1
30.0
25.7
21.5
25.8
21.5
a These data are the compiled results of studies done by three different
laboratories. These results are from samples spiked at 10 mg/kg.
b Not a PAH.
Data are taken from Reference 1.
8275A - 20
Revision 1
January 1995
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METHOD 8275A
SEMIVOLATILE ORGANIC COMPOUNDS (PAHs and PCBs)
IN SOILS/SLUDGES AND SOLID WASTES USING
THERMAL EXTRACTION/GAS CHROMATOGRAPHY/MASS SPECTROMETRY (TE/GC/MS)
7.1 Prepare crucible.
7.2.1 Sat the TE/GC/MS
system to recommended
conditions and bake out
the system.
7.2.2 Set the TE/GC/MS
system to recommended
conditions and calibrate.
7.2.3 Analyze method blank.
7.2.3 Make
appropriate
corrections.
.2.
Is the
system free
of target analytes
and inter-
ferences?
7.2.5 Prepare initial
calibration curve.
7.3 Verfiy calibration as
appropriate.
7.4.1 Prepare sample.
7.4.2 Determine sample
% dry weight.
7.4.5 Weigh internal
standard.
7.4.6 Weigh sample.
7.4.7 - 7.4.8 Load
sample into the thermal
extraction unit and analyze.
7.6 Calculate the cone.
of the analytes, or perform
a qualitative analysis.
(^ Stop J)
8275A - 24
Revision 1
January 1995
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METHOD 8515
COLORIMETRIC SCREENING METHOD FOR TRINITROTOLUENE (TNT) IN SOIL
1.0 SCOPE AND APPLICATION
1.1 Method 8515 is a procedure for screening soil samples to determine
when 2,4,6-trinitrotoluene (TNT, CAS No. 118-96-7) is present at concentrations
above 1 ppm.
1.2 Results obtained using this method should be used to locate samples
with TNT concentrations between 1 and 30 ppm. Extracts of samples reading >30
ppm should be diluted and re-evaluated in the test.
1.3 Using the test kit from which this method was developed, 95% of
samples containing 0.7 ppm of TNT or less will produce a negative result.
1.4 Method 8515 can be used to screen soil samples for the presence of TNT
and other chemically related nitroaromatic compounds e.g., dinitrotoluenes (DNT)
and 1,3,5-trinitrobenzene (TNB). It should be used as an indicator or screening
test for the presence of TNT. Method 8515 does not measure RDX or HMX.
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 Test kits are commercially available for this method. The
manufacturer's directions should be followed.
2.2 In general, the method is performed using an extract of a soil sample.
The sample is treated with color-change reagents and is read in a portable
spectrophotometer. The colorimetric nature of the test is based on the visual
detection of the reaction product that is formed when polynitroaromatic compounds
react with acetone by ketone substitution in the presence of base. This
substitution product is measured at 540 nm using a spectrophotometer. The
concentration of TNT in an unknown sample is determined by evaluating the
intensity of the color that is developed.
3.0 INTERFERENCES
3.1 Chemically similar compounds and compounds which might be expected to
be found in conjunction with TNT contamination were tested to determine the
concentration required to produce an equivalent TNT result. These data are shown
in Table 1.
8515 - 1 Revision 0
January 1995
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4.0 APPARATUS AND MATERIALS
4.1 TNT Soil Test System (EnSys, Inc.), or equivalent. Each commercially
available test kit will supply or specify the apparatus and materials necessary
for successful completion of the test.
4.2 UV/Vis Spectrophotometer, Hach DR/2000, or equivalent.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 See Section 6.0 of Method 8330.
6.3 Soil samples may be contaminated, and should therefore be considered
hazardous and handled accordingly.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance specifications indicated
in Tables 2-3.
7.2 Dry soil samples in air at room temperature or colder to a constant
weight, being careful not to expose the samples to direct sunlight.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for the quality control
procedures specific to the test kit being used. Additionally, guidance provided
in Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.3 Do not use test kits past their expiration dates.
8.4 Use the test kits within their specified storage temperature and
operating temperature limits.
8.5 Verify operation of the colorimeter/spectrophotometer by use of
appropriate standards.
8515 - 2 Revision 0
January 1995
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8.6 Method 8515 is intended for field or laboratory use. The appropriate
level of quality assurance should accompany the application of this method to
document data quality.
9.0 METHOD PERFORMANCE
9.1 Inter- and intra-assay precision data are provided in Tables 2 and 3,
respectively.
9.2 This method has been applied to a series of soil samples whose TNT
concentration had been determined by HPLC (Method 8330). These results are
provided in Table 4. A high degree of correlation was observed between the HPLC
method and the field method.
10.0 REFERENCES
1. T. F. Jenkins and M. W. Walsh, "Development of Field Screening Methods for
TNT, 2,4-DNT, and RDX in Soil", Talanta, 1992, 39 (4), 419-428.
2. T. F. Jenkins, "Development of a Simplified Field Method for the
Determination of TNT in Soil", Special Report 90-38 (November, 1990) USA
Cold Regions Research and Engineering Laboratory.
3. TNT Soil Test System Instructions for Use, EnSys, Inc.
11.0 SAFETY
11.1 Standard precautionary measures used for handling other organic
compounds should be sufficient for the safe handling of the samples, extracts and
standard solutions specified in this method. The only extra caution that should
be taken is when handling the analytical standard neat material. Follow Section
7.2 for drying the neat material at ambient temperature. If samples are taken
back to the laboratory for analysis by Method 8330, follow the additional safety
procedures specified in that method.
8515 - 3
Revision 0
January 1995
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TABLE 1
TNT Soil Test System
Sensitivity to Explosive Compounds
Compound
2,4,6-Trinitrotoluene
2,4-Dinitrotoluene
2,6-Dinitrotoluene
1, 3, 5-Tri nitrobenzene
2-Nitrotoluene
3-Nitrotoluene
4-Nitrotoluene
4-Amino-2,6-
dinitrotoluene
Nitrobenzene
Minimum Sensitivity
(ppm)
1
1.1
0.6
1
>100
>100
>100
>100
>100
TABLE 2
Inter-assay Precision of the TNT Soil Test
Spike
Concentration (ppm)
0
5
10
20
Average Result
(ppm ± SD)
0.0 ± 0.2
5.1 ± 0.4
10.1 ± 0.5
20.1 ± 0.8
%RSD
.
7.8%
4 . 5%
4.2%
These data were generated from 22 replicates in 10 matrices (9 soil
extracts and 1 acetone control).
8515 - 4
Revision 0
January 1995
-------
TABLE 3
Intra-assay Precision in the TNT Soil Test
Spike
Concentration (ppm)
0
10
Average Result
(ppm ± SD)
0.0 ± 0.2
10.2 ± 0.2
%RSD
-
1 . 9%
These data were generated from the 10 ppm TNT control provided with
the EnSys kit.
TABLE 4
Comparison of TNT Soil Test System
with Method 8330 (HPLC)
Sample
ID
012
028
022
021
023
024
027
025
026
016
013
015
020
019
Screening
Test Result
(ppm)
18.9
26.2
34
34.6
37.7
56.5
192
120
120
49
174
150
295
712
HPLC
Results
(ppm)
21.5
29.0
25.2
23.8
28.1
58.5
191
110
131
49
175
135
287
719
Does screening test
agree with HPLC
determination?
yes
yes
no
no
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
8515 - 5
Revision 0
January 1995
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METHOD 9078
SCREENING TEST METHOD FOR POLYCHLORINATED BIPHENYLS IN SOIL
1.0 SCOPE AND APPLICATION
1.1 The method may be used to determine the amount of PCB
(polychlorinated biphenyl) contamination in soils such as sand, gravel, loam,
sediment, and clay, assuming that PCBs are the sole source of halogens in the
sample.
1.2 This electrochemical method is designed to provide quantitative
field results over a range of 2 to 2000 jug/g PCBs, significantly cutting down
on the number of samples requiring laboratory testing.
1.3 Chlorines are removed from the PCB molecule using an organo-sodium
reagent. The resulting chloride ions are measured using a chloride specific
electrode. Analysts must identify the type of Aroclor contamination in order
to use this as a quantitative method.
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 A sample of the soil to be tested is extracted with a hydrocarbon
based solvent. The resulting extract is filtered to remove moisture and
inorganic salts. The dried extract is reacted with metallic sodium and a
catalyst to strip chloride from any PCB that may be present. The resulting
chloride ions are extracted into an aqueous buffer solution where they are
detected using a chloride ion specific electrode.
CAUTION: Some of the reagents used with this testing procedure contain
flammable solvents, dilute acids, and metallic sodium. Wear
gloves and safety glasses while performing tests. Read all MSDS
and warnings included with the instrument before starting testing
procedure.
3.0 INTERFERENCES
3.1 This procedure is sensitive to any chlorinated compound that is
preferentially soluble in a non-polar solvent. When analyzing for PCBs, the
presence of other chlorinated organics will result in a high bias. Iodine and
bromine containing compounds will affect results if present in significant
quantities. Wet or dry samples may be run, but results for all samples are
calculated on a wet-weight basis. In one evaluation study (Table 1), 10% of
the measurements were false negatives.
9078 - 1 Revision 0
January 1995
-------
3.2 Inorganic chlorides should not interfere using this method if the
sample is extracted with organic solvent.
4.0 APPARATUS AND MATERIALS
4.1 Electrochemical PCB test kit: L2000® PCB/Chloride Analyzer, (Dexsil
Corporation, One Hamden Park Drive, Hamden, CT), or equivalent. Each
commercially available test kit will supply or specify the apparatus and
materials necessary for successful completion of the test.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test. Reagents should be
labeled with appropriate expiration dates.
6.0 SAMPLE COLLECTION AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Soil samples may be contaminated, and should therefore be
considered hazardous and handled accordingly. All samples should be collected
using a sampling plan that addresses the considerations discussed in Chapter
Nine.
6.3 To achieve accurate analyses, soil samples should be well
homogenized prior to testing. PCBs are generally not evenly distributed in a
soil sample and extensive mixing must be done to assure consistency.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance specifications
indicated in Tables 1 and 2.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for quality control
procedures specific to the test kit used. Additionally, guidance provided in
Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.3 Method 9078 is intended for field or laboratory use. The
appropriate level of quality assurance should accompany the application of
this method to document data quality.
9078 - 2 Revision 0
January 1995
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9.0 METHOD PERFORMANCE
9.1 146 soil samples from a PCB contaminated site were analyzed. There
were 114 individual samples and 32 field duplicates. Each sample was analyzed
using both the L2000 and GC/MS. The L2000 analyses were performed on-site in
a mobile lab and the PCBs were analyzed as Aroclor 1242. Laboratory analyses
were performed on splits of the same samples. The results from the analyses
are presented in Table 1.
9.2 After applying accepted statistical methods to account for the
detection limit difference between the two methods the data were evaluated to
determine the acceptability of the L2000 method. A matched-pair students t-
test performed on the L2000 and CLP GC/MS data results in a t value of 0.2141.
This is well below the critical value (1.645 @ 0.05) for rejecting the null
hypothesis indicating that there is no statistical difference between the data
pairs. An analysis of the data for outliers identified only 2 data points
whose residuals were greater than 3 standard deviations (10 and 5
respectively). Both points were determined to be in error using other
evidence and were eliminated from the data set. A linear regression analysis
of the remaining data results in a correlation coefficient of 0.95 and a
positive intercept of 10.98 M9/9- The slope of 0.985 was not statistically
different from 1 and the intercept was not statistically different from 0.
9.3 The relative percent difference (RPD) calculated from all valid
duplicates greater than the L2000 detection limit of 2 /xg/g for each method
resulted in a mean RPD of 19% for the L2000 data and a mean RPD of 43% for the
CLP GC-MS method. A Dunnett's test shows that this is statistically
significant.
9.4 In a second study, soil samples contaminated with Aroclor 1260 were
taken during a site cleanup. The samples were split and sent for lab analysis
by Method 8082 as well as analysis by the L2000 in the field. The results are
reported in Table 2. A linear regression analysis of the data resulted in a
correlation coefficient of 0.995, a slope of 1.048 and an intercept of -1.48
/Ltg/g indicating that the L2000 is accurate compared to the lab method. A
calculation of the relative percent difference for data, where duplicates were
run within a method, results in a lower RPD for the L2000 indicating a tighter
data spread and better repeatability.
10.0 REFERENCE
1. Griffin, Roger D. Application of a New PCB Field Analysis Technique for
Site Assessment. Proceedings of Hazmacon '92 March - April 1992.
9078 - 3 Revision 0
January 1995
-------
TABLE 1
L2000 AND LABORATORY METHODS FROM STUDY #1
Summary of Results
Sample
Number
1
3
4
6
7
8
9
10
11
15
15D
16
17
18
19
23
25
32
33
34
36
38
40
43
43D
50
50D
52
53
54
55
59
60
60D
61
62
L2000
(M9/9)
ND
ND
23.6
ND
ND
3.9
6.9
5.1
2.7
9.4
12.5
484
6.5
382
71.1
48.8
3.5
36
ND
14.4
>2000
778
5.7
4.1
3.6
ND
ND
9.3
25.7
5.1
4.4
ND
2.3
4.4
549
111
GC/MS
(M9/9)
593
114
6.71
67
552
2
1.3
0.172
1.15
9.13
9.84
2110
2.55
45.4
6.7
20.8
11.7
47.6
6
34
816
1030
4.25
1.69
1.74
3.6
4.4
4.21
0.958
0.516
2.4
7.9
0.624
0.577
580
2.35
Results
Agree?
False Neg.
False Neg.
Yes
False Neg.
False Neg.
Yes
Yes
False Pos.
Yes
Yes
Yes
Yes
Yes
Yes
False Pos.
Yes
Yes
Yes
False Neg.
Yes
Yes
Yes
Yes
Yes
Yes
False Neg.
False Neg.
Yes
False Pos.
False Pos.
Yes
False Neg.
Yes
Yes
Yes
False Pos.
9078 - 4
Revision 0
January 1995
-------
Sample
Number
64
65
66
67
68
69
69D
73
74
75
76
78
79
80
84
84D
85
85D
88
88D
89
90
90D
91
91D
92
92D
95
95D
100
100D
101
102
102D
103
104
107
108
109
109D
L2000
Ug/g) _
172
ND
2.1
7.5
8
5.8
4.4
37
22
61
82
21
148
ND
7.6
10.9
593
596
ND
ND
ND
2
ND
1650
1608
3.14
3.4
20.6
20.1
384
363
8.3
6.3
5
75.2
4.1
161
6.1
P
10.3
GC/MS
(M9/9)
19
3.1
1.98
0.081
0.504
ND
ND
15.8
13.3
23
46.7
2.27
42.8
3.8
1.16
1.08
428
465
2.7
1.77
45
1.01
1.4
1630
1704
1.21
ND
17.5
31.2
177
167
1.21
293
1.77
40.3
7.66
14.1
3.84
ND
ND
Results
Agree?
Yes
False Neg.
Yes
False Pos.
False Pos.
False Pos.
False Pos.
Yes
Yes
Yes
Yes
Yes
Yes
False Neg.
Yes
False Pos.
Yes
Yes
False Neg.
Yes
False Neg.
Yes
Yes
Yes
Yes
Yes
False Pos.
yes
Yes
Yes
Yes
Yes
False Neg.
Yes
Yes
Yes
False Pos.
Yes
False Pos.
False Pos.
9078 - 5
Revision 0
January 1995
-------
Sample
Number
111
112
113
114
L2000
(M9/9)
20
240
21.8
107
GC/MS
(M9/9)
ND
315
14.9
66.3
Results
Agree?
False Pos.
Yes
Yes
Yes
79 out of 114 samples are reported in Table 1. Samples that were found
to be ND for both the L2000 kit and the GC/MS determination were not
reported. False negatives and positives were determined based on NDs
found by each technique relative to the detection limit of the L2000 kit
(2 M9/g)- False negatives and positives were also identified when
concentrations differed by more than an order of magnitude for results
from the two techniques.
ND = Not detected
15 False positives:
13 False negatives:
67 Non-detects:
ND - 14.1 ppm by GC/MS
2.7 - 593 ppm by GC/MS
ND - 2.5 ppm by GC/MS
TABLE 2
L2000 AND METHODS 8082 FROM STUDY #2
Summary of Results
Sample
Number
1
2
3
4
5
6
7
8
9
10
11
12
13
14
Method 8082
(M9/9)
83
21
12
300/375
29
106/134
3
9.3
1.5
99
7/9
3.6
4.2/6.2
290
L2000 Results
L_ (M9/9)
79/76
22
14
357/326/327
27
116/117
7.6
7.2
5.2
93
13
12
2.9
L_ 254/265
9078 - 6
Revision 0
January 1995
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METHOD 9079
SCREENING TEST METHOD FOR POLYCHLORINATED BIPHENYLS
IN TRANSFORMER OIL
1.0 SCOPE AND APPLICATION
1.1 Method 9079 may be used to screen hydrocarbon based electrical
insulating fluids for polychlorinated biphenyls (PCBs) at preset levels of 20,
50, 100, or 500 M9/9- The method is designed to provide screening data
outside of a laboratory environment in under 10 minutes, providing a
colorimetric indication that the concentration of PCBs is above or below the
fixed end point. Screening procedures may significantly reduce the number of
samples requiring laboratory testing.
1.2 Chlorines are removed from the PCB molecule using an organo-sodium
reagent. The resulting chloride ions are measured using a chloride specific
electrode. Analysts must identify the type of Aroclor contamination in order
to use this as a quantitative method.
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 A sample of the oil to be tested is reacted with a mixture of
metallic sodium catalyzed with naphthalene and diglyme at ambient temperature.
This process converts all organic halogens to their respective sodium halides.
All halides in the treated mixture, including those present prior to the
reaction, are then extracted into an aqueous buffer, a premeasured amount of
mercuric nitrate is added, followed by a solution of diphenylcarbazone as the
indicator.
2.2 The color of the solution at the end of the test indicates whether
the sample is above or below the preset chlorine level. A yellow end point
indicates a concentration greater than the set point of the test and a blue-
violet end point indicates a concentration less than the set point of the
test.
2.3 The end point at which each of the test kits turns positive is
calibrated using Aroclor 1242 standards. Aroclor 1242 provides a conservative
end point due to its low chlorine content relative to the other Aroclors used
in electrical equipment. A list of Aroclors used in electrical equipment and
the PCB concentration that gives a positive indication using the 50 /zg/g test
kit is given in Table 1.
9079 - 1 Revision 0
January 1995
-------
3.0 INTERFERENCES
3.1 Water present in the sample at more than 2% may cause a low
reading. Water present at this high a "level results in an obvious change in
the sodium reaction and the user should stop the test.
3.2 High sulfur levels (4%) will cause a high bias possibly resulting
in a false positive reading. The sample will also smell strongly of sulfur
after the sodium reaction.
3.3 Any chlorine contained in the sample will be measured as PCB
possibly resulting in a false positive if the total non-PCB chlorine
concentration in the sample is greater than the preset end point for the kit.
4.0 APPARATUS AND MATERIALS
4.1 Colorimetric test kit: Clor-N-Oil® (Dexsil Corporation, One Hamden
Park Drive, Hamden, CT), or equivalent. Each commercially available test kit
will supply or specify the apparatus and materials necessary for successful
completion of the test. Reagents should be labeled with appropriate
expiration dates.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 Soil and oil samples may be contaminated, and should therefore be
considered hazardous and handled accordingly. All samples should be collected
using a sampling plan that addresses the considerations discussed in Chapter
Nine.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance specifications
indicated in Tables 1-4.
CAUTION: Some of the reagents used with this testing procedure contain
flammable solvents, dilute acids, and metallic sodium. Wear
gloves and safety glasses while performing test. Read all MSDS
and warnings included with the kit before starting testing
procedure.
WARNING: Mercury waste must be properly disposed.
9079 - 2 Revision 0
January 1995
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8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for quality control
procedures specific to the test kit used. Additionally, guidance provided in
Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.6 Method 9079 is intended for field or laboratory use. The
appropriate level of quality assurance should accompany the application of
this method to document data quality.
9.0 METHOD PERFORMANCE
9.1 A double blind study was conducted using the Clor-N-Oil 50 on 6
spiked transformer oil samples. The spiking concentrations were chosen to be
identical to those from three sets of the EPA's Water Pollution (WP)
Performance Evaluation Program. They were made in Shell Diala A electrical
insulating fluid at the concentrations listed in Table 1.
9.1.1 A total of 38 operators participated in the test, 10 of
whom had prior experience with the test method, and 27 of whom had no
prior experience using the kits, and 1 of whom had viewed a test
demonstration before running the test themselves. This distribution of
operators was chosen to verify the robust nature of the method in light
of the vast range of experience typical of users in the field.
9.1.2 Each operator was given six random samples containing an
unknown concentration of PCBs. Operators recorded their results as
greater than or less than 50 M9/9- Only after all of the tests were
run were the data collected and compared with the known values.
9.1.3 Out of the 228 tests run, 4 were invalid due to spillage of
reagents or improper kit operation resulting in an incomplete test. The
test data are presented in Table 2. From these data, it is evident that
there is a much higher likelihood of obtaining a false positive reading
than a false negative.
9.1.4 The expected certainties estimated from these data are
presented in Table 3. The likelihood of obtaining a false positive
approaches 90% at 90% of the action level. This reflects the
conservative design of the test. At the action level of 50 M9/9>
nearly 99% of the samples would be identified correctly as containing 50
/ug/g of PCB or greater. These results represent errors due to all
sources and therefore represent real world performance of the method by
field personnel.
9.1.5
ppm.
ie i .
For samples containing 45 /ig/g, 37 of 41 gave results >50
9079 - 3 Revision 0
January 1995
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9.2 The real world performance was investigated in detail by Utah Power
& Light Company. They tested the insulating fluid from approximately 200,000
pieces of electrical equipment with the Clor-N-Oil 50 test kit in order to
classify them as either PCB or non-PCB (i.e., containing more or less than
50 jzg/g)- Each piece of equipment was tested once with the test kit. A
random sample of 937 of the Clor-N-Oil negatives were tested by Gas
Chromatography to confirm the results. The results from the testing are
summarized in Table 4. The false negative rate predicted from the data is
less than 1% (0.65%). This means that a transformer can be classified as
non-PCB with greater than 99% confidence.
10.0 REFERENCES
1. Finch, S.; Lynn, T.B; Lynn T.D.; and Scott, R.P.W., Which Method is the
Most Reliable in the Field, EPRI PCB seminar Proceedings October 1991.
2. Mills, David W.; and Rhoads, Kirt W., Clor-N-Oil Test Kit as a PCB
Screening Tool 1985. EPRI PCB Seminar Proceedings March 1986. EPRI
CS/EA/EL-4480 Product 2028.
3. Rhoads, Kirt W., Clor-N-Oil Test Kit as a Risk Management Tool - An
Update 1987. EPRI PCB Seminar Proceedings December 1987.
f
9079 - 4
Revision 0
January 1995
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TABLE 1
SPIKING CONCENTRATIONS FOR Clor-N-Oil
PCB Concentrations in Shell Diala A
Aroclor
1242
1242
1254
1254
1260
1260
Concentration
21.2 M9/9
45.0 Mg/g
26.3 /xg/g
50.0 //g/g
8.2 M9/9
50.0 M9/9
TABLE 2
ESTIMATED ERROR RATES FOR Clor-N-Oil 50
PCB Concentration
M9/9
8
21
26
45
50
Expected False
Positive Rate %
<3
2.8
35
88
—
Expected False
Negative Rate %
—
—
—
—
1.4
TABLE 3
GC RETEST RESULTS OF 930 NEGATIVES CLASS INTERVAL (ng/g)
Number
Percent
0-1
768
82.6
1-5
93
10
6-15
48
5.2
16-25
5
0.5
26-46
10
1.1
47-99
3
0.3
100-475
3
0.3
476-999
0
0
1000+
0
0
9079 - 5
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TABLE 4
TEST RESULTS FOR Clor-N-Oil 50 STUDY
Sample
ID
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
Concentration
PCB (Mg/g)
50
26.3
50
45
21
8.2
21
45
26.3
50
8.2
50
26.3
45
8.2
45
50
50
8.2
26.3
45
21
45
50
21
50
50
Aroclor
1254
1254
1260
1242
1242
1260
1242
1242
1254
1254
1260
1260
1254
1242
1260
1242
1260
1254
1260
1254
1242
1242
1242
1260
1242
1260
1254
Field Test
Result (jug/g)
>50
<50
NA
>50
<50
<50
<50
<50
<50
>50
<50
>50
<50
>50
<50
>50
>50
>50
<50
<50
>50
<50
<50
>50
<50
>50
>50
9079 - 6
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Sample
ID
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
Concentration
PCB (/xg/g)
21
45
50
8.2
50
50
50
45
26.3
50
26.3
50
21
50
8.2
26.3
8.2
21
26.3
8.2
21
50
50
8.2
45
21
26.3
21
45
8.2
Aroclor
1242
1242
1254
1260
1254
1260
1254
1242
1254
1254
1254
1260
1242
1260
1260
1254
1260
1242
1254
1260
1242
1260
1260
1260
1242
1242
1254
1242
1242
1260
Field Test
Result (jug/g)
<50
>50
>50
<50
>50
>50
>50
>50
<50
>50
<50
>50
<50
>50
<50
<50
<50
<50
<50
<50
<50
>50
>50
<50
<50
<50
<50
<50
>50
<50
9079 - 7
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Sample
ID
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
Concentration
PCB (Mg/g)
50
26.3
21
8.2
45
50
50
50
50
21
50
50
45
26.3
21
8.2
8.2
50
45
45
21
8.2
50
50
26.3
26.3
26.3
21
26.3
45
Aroclor
1254
1254
1242
1260
1242
1254
1254
1254
1260
1242
1260
1260
1242
1254
1242
1260
1260
1260
1242
1242
1242
1260
1254
1254
1254
1254
1254
1242
1254
1242
Field Test
Result (/Ltg/g)
>50
>50
<50
<50
>50
>50
>50
>50
>50
<50
>50
>50
<50
<50
<50
<50
<50
>50
<50
>50
<50
<50
>50
>50
<50
<50
<50
<50
<50
>50
9079 - 8
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Sample
ID
88
89
90
91
92
93
97
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
Concentration
PCB (Mg/g)
50
8.2
50
45
8.2
50
45
21
50
45
45
45
26.3
50
26.3
26.3
21
26.3
50
8.2
21
26.3
50
21
45
26.3
50
21
21
26.3
Aroclor
1254
1260
1260
1242
1260
1260
1242
1242
1254
1242
1242
1242
1254
1260
1254
1254
1242
1254
1260
1260
1242
1254
1254
1242
1242
1254
1260
1242
1242
1254
Field Test
Result (MQ/Q)
>50
<50
>50
>50
<50
>50
>50
<50
>50
>50
>50
>50
>50
>50
>50
<50
<50
>50
>50
<50
<50
<50
>50
<50
>50
<50
>50
<50
<50
<50
9079 - 9
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Sample
ID
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
146
147
Concentration
PCB (Mg/g)
8.2
50
50
50
26.3
45
50
8.2
50
8.2
45
50
21
26.3
50
26.3
45
26.3
50
50
45
21
50
8.2
8.2
50
8.2
45
8.2
26.3
Aroclor
1260
1254
1254
1254
1254
1242
1260
1260
1254
1260
1242
1260
1242
1254
1260
1254
1242
1254
1260
1260
1242
1242
1254
1260
1260
1260
1260
1242
1260
1254
Field Test
Result (jug/g)
<50
>50
>50
>50
>50
>50
>50
<50
>50
<50
>50
>50
<50
<50
>50
<50
>50
<50
>50
>50
>50
<50
>50
<50
<50
>50
<50
>50
<50
<50
9079 - 10
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Sample
ID
148
149
150
151
152
153
154
155
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
176
177
Concentration
PCB (/ig/g)
21
8.2
21
50
8.2
50
50
21
50
26.3
21
26.3
8.2
50
45
50
50
50
21
50
50
45
50
50
21
50
8.2
45
8.2
8.2
Aroclor
1242
1260
1242
1254
1260
1254
1254
1242
1260
1254
1242
1254
1260
1254
1242
1254
1254
1260
1242
1254
1260
1242
1260
1260
1242
1260
1260
1242
1260
1260
Field Test
Result (/Ltg/g)
<50
<50
<50
>50
<50
>50
>50
NA
NA
<50
<50
>50
<50
>50
>50
>50
>50
>50
<50
>50
>50
>50
<50
>50
<50
>50
<50
>50
<50
<50
9079 - 11
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January 1995
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Sample
ID
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
201
202
203
204
205
206
207
Concentration
PCB (Mg/g)
45
26.3
50
45
45
21
26.3
50
21
50
50
26.3
8.2
21
8.2
45
8.2
21
26.3
50
50
26.3
45
21
8.2
21
45
50
50
26.3
Aroclor
1242
1254
1254
1242
1242
1242
1254
1254
1242
126
1254
1254
1260
1242
1260
1242
1260
1242
1254
1260
1260
1254
1242
1242
1260
1242
1242
1254
1254
1254
Field Test
Result (jug/g}
>50
>50
>50
>50
>50
<50
>50
>50
<50
>50
>50
<50
<50
<50
<50
>50
<50
<50
>50
>50
>50
>50
>50
>50
<50
<50
>50
>50
>50
NA
9079 - 12
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Sample
ID
208
209
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
226
227
228
Concentration
PCB (Mg/g)
50
50
50
45
21
50
50
8.2
8.2
50
45
26.3
26.3
21
45
45
45
26.3
8.2
45
50
Aroclor
1254
1254
1260
1242
1242
1260
1260
1260
1260
1260
1242
1254
1254
1242
1242
1242
1242
1254
1260
1242
1254
Field Test
Result Ug/g)
>50
>50
>50
>50
<50
>50
>50
<50
<50
>50
>50
>50
>50
>50
>50
>50
>50
>50
<50
>50
>50
9079 - 13
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CHAPTER FIVE
MISCELLANEOUS TEST METHODS
The following methods are found in Chapter Five:
Method 5050:
Method 9010A:
Method 9012A:
Method 9013:
Method 9020B:
Method 9021:
Method 9022:
Method 9023:
Method 9030A:
Method 9031:
Method 9035:
Method 9036:
Method 9038:
Method 9056:
Method 9057:
Method 9060:
Method 9065:
Method 9066:
Method 9067:
Method 9070:
Method 9071A:
Method 9075:
Method 9076:
Bomb Preparation Method for Solid Waste
Total and Amenable Cyanide (Colorimetric, Manual)
Total and Amenable Cyanide (Colorimetric,
Automated UV)
Cyanide Extraction Procedure for Solids and Oils
Total Organic Hal ides (TOX)
Purgeable Organic Hal ides (POX)
Total Organic Hal ides (TOX) by Neutron Activation
Analysis
Extractable Organic Hal ides (EOX) in Solids
Acid-Soluble and Acid-Insoluble Sulfides
Extractable Sulfides
Sulfate (Colorimetric, Automated, Chloranilate)
Sulfate (Colorimetric, Automated, Methylthymol
Blue, AA II)
Sulfate (Turbidimetric)
Determination of Inorganic Anions
Chromatography
Determination of Chloride from HC1/HCL
Sampling Train (Methods 0050 and 0051)
Chromatography
Total Organic Carbon
Phenolics (Spectrophotometric, Manual 4-AAP
Distillation)
Phenolics (Colorimetric, Automated 4-AAP with
Distillation)
Phenolics (Spectrophotometric,
Distillation)
Total Recoverable Oil & Grease
Separatory Funnel Extraction)
Oil and Grease Extraction Method
Sediment Samples
Chlorine in New and Used
by X-Ray Fluorescence
by Ion
Emission
by An ion
with
MBTH with
(Gravimetric,
for Sludge and
Test Method for Total
Petroleum Products
Spectrometry (XRF)
Test Method for Total
Petroleum Products by
Microcoulometry
Chlorine in New and Used
Oxidative Combustion and
FIVE - 1
Revision 2
January 1995
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Method 9077:
Method A:
Method B:
Method C:
Method 9131:
Method 9132:
Method 9210:
Method 9211:
Method 9212:
Method 9213:
Method 9214:
Method 9215:
Method 9250:
Method 9251:
Method 9253:
Method 9320:
Test Methods for Total Chlorine in New and Used
Petroleum Products (Field Test Kit Methods)
Fixed End Point Test Kit Method
Reverse Titration Quantitative End Point
Test Kit Method
Direct Titration Quantitative End Point Test Kit
Method
Total Coliform: Multiple Tube Fermentation
Technique
Total Coliform: Membrane Filter Technique
Potentiometric Determination of Nitrate in
Aqueous Samples with Ion-Selective Electrode
Potentiometric Determination of Solubilized
Bromide in Aqueous Samples with Ion-Selective
Electrode
Potentiometric Determination of
Aqueous Samples with Ion-Selective
Potentiometric Determination
Cyanide in Aqueous Samples and
Ion-Selective Electrode
Potentiometric Determination of Fluoride in
Aqueous Samples with Ion-Selective Electrode
Potentiometric Determination of Sulfide in
Aqueous Samples and Distillates with Ion-
Selective Electrode
Chloride (Colorimetric, Automated Ferricyanide AAI)
Chloride (Colorimetric, Automated Ferricyanide
AAI I)
Chloride (Titrimetric, Silver Nitrate)
Radium-228
Chloride in
Electrode
of Solubilized
Distillates with
FIVE - 2
Revision 2
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METHOD 9012A
TOTAL AND AMENABLE CYANIDE (COLORIMETRIC, AUTOMATED UV)
1.0 SCOPE AND APPLICATION
1.1 Method 9012 is used to determine the concentration of inorganic
cyanide (CAS Registry Number 57-12-5) in wastes or leachate. The method detects
inorganic cyanides that are present as either soluble salts or complexes. It is
used to determine values for both total cyanide and cyanide amenable to
chlorination. The "reactive" cyanide content of a waste, that is, the cyanide
content that could generate toxic fumes when exposed to mild acidic conditions,
is not distilled by Method 9012 (refer to Chapter Seven). However, Method 9012
may be used to quantify the concentration of cyanide from the reactivity test.
2.0 SUMMARY OF METHOD
2.1 The cyanide, as hydrocyanic acid (HCN), is released from samples
containing cyanide by means of a reflux-distillation operation under acidic
conditions and absorbed in a scrubber containing sodium hydroxide solution. The
cyanide ion in the absorbing solution is then determined by automated UV
colorimetry.
2.2 In the automated colorimetric measurement, the cyanide is converted
to cyanogen chloride (CNC1) by reaction with Chloramine-T at a pH less than 8
without hydrolyzing to the cyanate. After the reaction is complete, color is
formed on the addition of pyridine-barbituric acid reagent. The concentration
of NaOH must be the same in the standards, the scrubber solutions, and any
dilution of the original scrubber solution to obtain colors of comparable
intensity.
3.0 INTERFERENCES
3.1 Interferences are eliminated or reduced by using the distillation
procedure. Chlorine and sulfide are interferences in Method 9012.
3.2 Oxidizing agents such as chlorine decompose most cyanides.
Chlorine interferences can be removed by adding an excess of sodium arsenite to
the waste prior to preservation and storage of the sample to reduce the chlorine
to chloride which does not interfere.
3.3 Sulfide interference can be removed by adding an excess of bismuth
nitrate to the waste (to precipitate the sulfide) before distillation. Samples
that contain hydrogen sulfide, metal sulfides, or other compounds that may
produce hydrogen sulfide during the distillation should be treated by the
addition of bismuth nitrate.
3.4 High results may be obtained for samples that contain nitrate
and/or nitrite. During the distillation, nitrate and nitrite will form nitrous
acid, which will react with some organic compounds to form oximes. These
9012A - 1 Revision 1
January 1995
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compounds once formed will decompose under test conditions to generate HCN. The
possibility of interference of nitrate and nitrite is eliminated by pretreatment
with sulfamic acid just before distillation. Nitrate and nitrite are
interferences when present at levels higher than 10 mg/L and in conjunction with
certain organic compounds.
3.5 Thiocyanate is reported to be an interference when present at very
high levels. Levels of 10 mg/L were not found to interfere in Method 9010.
3.6 Fatty acids, detergents, surfactants, and other compounds may cause
foaming during the distillation when they are present in large concentrations and
will make the endpoint of the titration difficult to detect. They may be
extracted at pH 6-7.
4.0 APPARATUS AND MATERIALS
4.1 Reflux distillation apparatus such as shown in Figure 1 or Figure
2. The boiling flask should be of one liter size with inlet tube and provision
for condenser. The gas scrubber may be a 270-mL Fisher-Milligan scrubber
(Fisher, Part No. 07-513 or equivalent). The reflux apparatus may be a Wheaton
377160 distillation unit or equivalent.
4.2 Automated continuous-flow analytical instrument with:
4.2.1 Sampler.
4.2.2 Manifold.
4.2.3 Proportioning pump.
4.2.4 Heating bath with distillation coil.
4.2.5 Distillation head.
4.2.6 Colorimeter equipped with a 15-mm flowcell and 570 nm
filter.
4.2.7 Recorder.
4.3 Hot plate stirrer/heating mantle.
4.4 pH meter.
4.5 Amber light.
4.6 Vacuum source.
4.7 Refrigerator.
4.8 5 mL microburette.
9012A - 2 Revision 1
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4.9 7 Class A volumetric flasks - 100 and 250 ml.
4.10 Erlenmeyer flask - 500 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 reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Reagents for sample collection, preservation, and handling
5.3.1 Sodium arsenite (0.1N), NaAs02. Dissolve 3.2 g NaAs02 in
250 ml water.
5.3.2 Ascorbic acid, C6H806.
5.3.3 Sodium hydroxide solution (50%), NaOH. Commercially
available.
5.3.4 Acetic acid (1.6M) CH3COOH. Dilute one part of
concentrated acetic acid with 9 parts of water.
5.3.5 2,2,4-Trimethylpentane, C8H18.
5.3.6 Hexane, C6H14.
5.3.7 Chloroform, CHC13.
5.4 Reagents for cyanides amenable to chlorination
5.4.1 Calcium hypochlorite solution (0.35M), Ca(OCl)2. Combine
5 g of calcium hypochlorite and 100 ml of water. Shake before using.
5.4.2 Sodium hydroxide solution (1.25N), NaOH. Dissolve 50 g
of NaOH in 1 liter of water.
5.4.3 Sodium arsenite (0.1N). See Step 5.3.1.
5.4.4 Potassium iodide starch paper.
9012A - 3 Revision 1
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5.5 Reagents for distillation
5.5.1 Sodium hydroxide (1.25N). See Step 5.4.2.
5.5.2 Bismuth nitrate (0.062M), Bi(NO)3 • 5H20. Dissolve 30 g
Bi(NO)3 • 5H20 in 100 ml of water. While stirring, add 250 ml of glacial
acetic acid, CH3COOH. Stir until dissolved and dilute to 1 liter with
water.
5.5.3 Sulfamic acid (0.4N), H2NS03H. Dissolve 40 g H2NS03H in
1 liter of water.
5.5.4 Sulfuric acid (18N), H2S04. Slowly and carefully add 500
mL of concentrated H2S04 to 500 ml of water.
5.5.5 Magnesium chloride solution (2.5M), MgCl2» 6H20. Dissolve
510 g of MgCl2 • 6H20 in 1 liter of water.
5.5.6 Lead acetate paper.
5.6 Reagents for automated colorimetric determination
5.6.1 Pyridine-barbituric acid reagent: Place 15 g of
barbituric acid in a 250-mL volumetric flask, add just enough reagent
water to wash the sides of the flask, and wet the barbituric acid. Add 75
ml of pyridine and mix. Add 15 ml of concentrated HC1, mix, and cool to
room temperature. Dilute to 250 mL with reagent water and mix. This
reagent is stable for approximately six months if stored in a cool, dark
place.
5.6.2 Chloramine-T solution: Dissolve 2.0 g of white, water
soluble chloramine-T in 500 mL of reagent water and refrigerate until
ready to use.
5.6.3 Sodium hydroxide, 1 N: Dissolve 40 g of NaOH in reagent
water, and dilute to 1 liter.
5.6.4 All working standards should contain 2 mL of 1 N NaOH
(Step 5.6.3) per 100 mL.
5.6.5 Dilution water and receptacle wash water (NaOH, 0.25 N):
Dissolve 10.0 g NaOH in 500 mL of reagent water. Dilute to 1 liter.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine.
6.2 Samples should be collected in plastic or glass containers. All
containers must be thoroughly cleaned and rinsed.
9012A - 4 Revision 1
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6.3 Oxidizing agents such as chlorine decompose most cyanides. To
determine whether oxidizing agents are present, test a drop of the sample with
potassium iodide-starch test paper. A blue color indicates the need for
treatment. Add 0.1N sodium arsenite solution a few mL at a time until a drop of
sample produces no color on the indicator paper. Add an additional 5 ml of
sodium arsenite solution for each liter of sample. Ascorbic acid can be used as
an alternative although it is not as effective as arsenite. Add a few crystals
of ascorbic acid at a time until a drop of sample produces no color on the
indicator paper. Then add an additional 0.6 g of ascorbic acid for each liter
of sample volume.
6.4 Aqueous samples must be preserved by adding 50% sodium hydroxide
until the pH is greater than or equal to 12 at the time of collection.
6.5 Samples should be chilled to 4°C.
6.6 When properly preserved, cyanide samples can be stored for up to
14 days prior to sample preparation steps.
6.7 Solid and oily wastes may be extracted prior to analysis by Method
9013 (Cyanide Extraction Procedure for Solids and Oils). It uses a dilute NaOH
solution (pH = 12) as the extractant. This yields extractable cyanide.
6.8 If fatty acids, detergents, and surfactants are a problem, they may
be extracted using the following procedure. Acidify the sample with acetic acid
(1.6M) to pH 6.0 to 7.0.
CAUTION: This procedure can produce lethal HCN gas.
Extract with isooctane, hexane, or chloroform (preference in order named) with
solvent volume equal to 20% of the sample volume. One extraction is usually
adequate to reduce the compounds below the interference level. Avoid multiple
extractions or a long contact time at low pH in order to keep the loss of HCN at
a minimum. When the extraction is completed, immediately raise the pH of the
sample to above 12 with 50% NaOH solution.
7.0 PROCEDURE
7.1 Pretreatment for cyanides amenable to chlorination
7.1.1 This test must be performed under amber light. K3[Fe-
(CN)6] may decompose under UV light and hence will test positive for
cyanide amenable to chlorination if exposed to fluorescent lighting or
sunlight. Two identical sample aliquots are required to determine cyanides
amenable to chlorination.
7.1.2 To one 500 ml sample or to a sample diluted to 500 ml, add
calcium hypochlorite solution dropwise while agitating and maintaining the
pH between 11 and 12 with 1.25N sodium hydroxide until an excess of
9012A - 5 Revision 1
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chlorine is present as indicated by Kl-starch paper turning blue. The
sample will be subjected to alkaline chlorination by this step.
CAUTION: The initial reaction product of alkaline chlorination is
the very toxic gas cyanogen chloride; therefore, it is necessary
that this reaction be performed in a hood.
7.1.3 Test for excess chlorine with Kl-starch paper and maintain
this excess for one hour with continuous agitation. A distinct blue color
on the test paper indicates a sufficient chlorine level. If necessary,
add additional calcium hypochlorite solution.
7.1.4 After one hour, add 1 ml portions of 0.1N sodium arsenite
until Kl-starch paper shows no residual chlorine. Add 5 ml of excess
sodium arsenite to ensure the presence of excess reducing agent.
7.1.5 Test for total cyanide as described below in both the
chlorinated and the unchlorinated samples. The difference of total
cyanide in the chlorinated and unchlorinated samples is the cyanide
amenable to chlorination.
7.2 Distillation Procedure
7.2.1 Place 500 ml of sample, or sample diluted to 500 ml in the
one liter boiling flask. Pipet 50 ml of 1.25N sodium hydroxide into the
gas scrubber. If the apparatus in Figure 1 is used, add water until the
spiral is covered. Connect the boiling flask, condenser, gas scrubber and
vacuum trap.
7.2.2 Start a slow stream of air entering the boiling flask by
adjusting the vacuum source. Adjust the vacuum so that approximately two
bubbles of air per second enter the boiling flask through the air inlet
tube.
7.2.3 If samples are known or suspected to contain sulfide, add
50 mL of 0.062M bismuth nitrate solution through the air inlet tube. Mix
for three minutes. Use lead acetate paper to check the sample for the
presence of sulfide. A positive test is indicated by a black color on the
paper.
7.2.4 If samples are known or suspected to contain nitrate or
nitrite, or if bismuth nitrate was added to the sample, add 50 ml of 0.4N
sulfamic acid solution through the air inlet tube. Mix for three minutes.
Note: Excessive use of sulfamic acid could create method bias.
7.2.5 Slowly add 50 mL of 18N sulfuric acid through the air
inlet tube. Rinse the tube with water and allow the airflow to mix the
flask contents for three minutes. Add 20 ml of 2.5M magnesium chloride
through the air inlet and wash the inlet tube with a stream of water.
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7.2.6 Heat the solution to boiling. Reflux for one hour. Turn
off heat and continue the airflow for at least 15 minutes. After cooling
the boiling flask, and closing the vacuum source, disconnect the gas
scrubber.
7.2.7 Transfer the solution from the scrubber into a 250-mL
volumetric flask. Rinse the scrubber into the volumetric flask. Dilute
to volume with water.
7.3 Automated colorimetric determination:
7.3.1 Set up the manifold in a hood or a well-ventilated area
as shown in Figure 3.
7.3.2 Allow colorimeter and recorder to warm up for 30 min. Run
a baseline with all reagents, feeding reagent water through the sample
line.
7.3.3 Place appropriate standards in the sampler in order of
increasing concentration. Complete loading of the sampler tray with
unknown samples.
7.3.4 When the baseline becomes steady, begin the analysis.
7.4 Standard curve for samples without sulfide
7.4.1 Prepare a series of standards by pipetting suitable
volumes of working standard potassium cyanide solution into 250-mL
volumetric flasks. To each flask, add 50 mL of 1.25N sodium hydroxide and
dilute to 250 ml with water. Prepare using the following table. The
sodium hydroxide concentration will be 0.25N.
ml of Working Standard Solution Concentration
(1 mL = 10 uq CN") (uq CNVL1
0 Blank
1.0 40
2.0 80
5.0 200
10.0 400
15.0 600
20.0 800
7.4.2 After the standard solutions have been prepared according
to the table above, pipet 50 mL of each standard solution into a 100-mL
volumetric flask and proceed to Steps 7.3.2 and 7.3.3 to obtain absorbance
values for the standard curve. The final concentrations for the standard
curve will be one half of the amounts in the above table (final
concentrations ranging from 20 to 400 ug/L).
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7.4.3 It is recommended that at least two standards (a high and
a low) be distilled and compared to similar values on the curve to ensure
that the distillation technique is reliable. If distilled standards do
not agree within + 10% of the undistilled standards, the analyst should
find the cause of the apparent error before proceeding.
7.4.4 Prepare a standard curve ranging from 20 to 400 jug/L by
plotting absorbance of standard versus the cyanide concentration
7.5 Standard curve for samples with sulfide
7.5.1 It is imperative that all standards be distilled in the
same manner as the samples using the method of standard additions.
Standards distilled by this method will give a linear curve, at low
concentrations, but as the concentration increases, the recovery
decreases. It is recommended that at least five standards be distilled.
7.5.2 Prepare a series of standards similar in concentration to
those mentioned in Step 7.4.1 and analyze as in Step 7.3. Prepare a
standard curve by plotting absorbance of standard versus the cyanide
concentration.
7.6 Calculation: Prepare a standard curve by plotting peak heights of
standards against their concentration values. Compute concentrations of samples
by comparing sample peak heights with the standard curve.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Verify the calibration curve with an independent calibration check
standard. If the standards are not within 15% of the expected value, a new
recalibration curve is required. Verify the calibration curve with every sample
batch by analyzing a mid-range standard.
8.3 Run one matrix spike sample for every 10 samples to check the
efficiency of sample distillation. A matrix spike should be prepared by adding
cyanide from the working standard or intermediate standard to 500 mL of sample
to ensure a concentration of approximately 40 /xg/L. Both the matrix duplicate
and matrix spike duplicate are brought through the entire sample preparation and
analytical process.
8.4 The method of standard additions shall be used for the analysis of
all samples that suffer from matrix interferences such as samples which contain
sulfides.
9.0 METHOD PERFORMANCE
9.1 Precision and accuracy data are not available at this time.
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10.0 REFERENCES
1. Annual Book of ASTM Standards, Part 31, "Water," Standard D2036-75, Method
B, p. 505 (1976).
2. Goulden, P.O., B.K. Afghan, and P. Brooksbank, Determination of Nanogram
Quantities of Simple and Complex Cyanides in Water, Anal. Chem., 44(11), pp.
1845-49 (1972).
3. Standard Methods for the Examination of Water and Wastewater, 14th ed., pp.
376 and 370, Method 413F and D (1975).
4. Technicon AutoAnalyzer II Methodology, Industrial Method No. 315-74 WCUV
Digestion and Distillation, Technicon Industrial Systems, Tarrytown, New York,
10591 (1974).
9012A - 9 Revision 1
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Connecting Tubing
Allihn Condenser
Air Inlet Tube
One-Liter
Boiling Flask
Suction
Figure 1. Apparatus for Cyanide Distillation
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Revision 1
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Cooling Water
Inlet Tube *
Screw Clamp
I
Heater •»
To Low Vacuum Source
Gas Scrubber
Condenser
Distilling Flask
O
Figure 2. Cyanide Distillation Apparatus
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a
i
o
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METHOD 9012A
TOTAL AND AMENABLE CYANIDE fCOLQRIMETRIC. AUTOMATED UV)
7.1 Pretreat to
determine
cyanides amenable
to chlormation.
7.2.1 Place sample
in flask; pipet
sodium hydroxide
into absorbing
tube.
7.2.4
Are samples
suspected to
contain NO2
and/or
No3?
7.2.4 Add
sulfamic
acid solution
through air
inlet tube.
7.2.2 Introduce
air stream into
boiling flask.
7.2.5 Add
rinse tube with
Type II water
add magnesium
chloride.
7.2.3
Positive
sulfide
test?
7.2.3 Treat
sample by
adding bismuth
nitrate solution.
7.2.6 Boil
solution;
reflux; cool;
close off
vacuum source.
7.2.7 Drain
solution
from absorber
into flask.
7.3 Perform
baseline
colorimetric
analysis.
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METHOD 9012A
TOTAL AND AMENABLE CYANIDE (COLORIMETRIC. AUTOMATED UV)
(CONTINUED)
7.5.1 Distill
standards in
same manner
as sample.
7.4
Does
sample
contain
sulfide?
7.4.1 Prepare a
series of
CN standards.
7.5.2 Prepare
standard curve
of absorbances.
7.4.2 Distill at
least two
standards to check
distillation
techniques.
7.4.3 Prepare
standard curve
of absorbances.
7.6 Compute
concentrations.
7.4.4 Check
efficiency of
sample
distillation.
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METHOD 9023
EXTRACTABLE ORGANIC HALIDES (EOX) IN SOLIDS
1.0 SCOPE AND APPLICATION
1.1 This method is to be used for the determination of total extractable
organic halides (EOX) as CV in solids. EOX is defined as the sum of those
organic halides which are extracted and detected by pyrolysis/microcoulometry
under the conditions specified in this method. Extractable organic halides
containing chlorine, bromine, or iodine are detected. However, fluorine
containing species are not detected by this method.
1.2 This method has been evaluated for solid wastes, soils, and suspended
solids isolated from industrial wastewater.
1.3 This method is recommended for use in the concentration range from
the MDL up to 1000 x MDL (see Section 9.1).
1.4 This method is restricted to use by, or under the supervision of,
analysts experienced in the operation of a pyrolysis microcoulometer and in the
interpretation of the results.
1.5 Since this method does not identify individual components, it is
advisable that compound specific techniques be employed to determine the
individual components present in samples exhibiting significant EOX levels,
unless the nature of the sample is already known.
2.0 SUMMARY OF METHOD
2.1 A 1-gram aliquot of solid sample is extracted with ethyl acetate by
sonification to isolate organic halides. A 25 juL aliquot of the extract is
either injected or delivered by boat inlet into a pyrolysis furnace using a
stream of C02/02 and the hydrogen halide (HX) pyrolysis product is determined by
microcoulometric titration.
3.0 INTERFERENCES
3.1 Method interferences may be caused by contaminants, reagents,
glassware, and other sample processing hardware. All of these materials must be
routinely demonstrated to be free from interferences under the conditions of the
analysis by running method blanks.
3.1.1 Glassware must be scrupulously cleaned. Clean all glassware
as soon as possible after use by treating with chromate cleaning solution.
This should be followed by detergent washing in hot water. Rinse with tap
water and distilled water, drain dry, and heat in a muffle furnace at
400°C for 15 to 30 minutes. Volumetric ware should not be heated in a
muffle furnace. Glassware should be sealed and stored in a clean
9023 - 1 Revision 0
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environment after drying and cooling to prevent any accumulation of dust
or other contaminants.
3.1.2 The use of high purity reagents and gases helps to
minimize interference problems.
3.1.3 The use of non-TFE (polytetrafluoroethylene) plastic tubing,
non-TFE thread sealants, or flow controllers with rubber components in the
purge gas stream should be avoided.
3.2 Samples can be contaminated by diffusion of volatile organics
(particularly solvents such as methylene chloride) through the septum seal into
the sample during shipment and storage.
3.3 All operations should be carried out in an area where halogenated
solvents, such as methylene chloride, are not being used.
3.4 Certain inorganic halide salts (e.g.. mercuric chloride) will be
extracted, and therefore interfere to some extent.
4.0 APPARATUS AND MATERIALS
4.1 Modified Dohrmann microcoulometric-titration system DX-20, or
equivalent, containing the following components:
4.1.1 Solvent injection system.
4.1.2 Pyrolysis furnace.
4.1.3 Titration cell.
4.2 Boat inlet or Microsyringes - 10, 25 /nL with 26 gauge 4-inch-long
needle.
4.3 Laboratory centrifuge to hold 15 mL conical centrifuge tubes.
4.4 Sonic bath or sonic probe to fit 10 mL vial. A power level of at
least 200 watts is required.
4.5 Centrifuge Tubes - 15 mL, conical, with Teflon*-lined screw caps.
4.6 Vials - 10 mL, with Teflon*-!ined screw caps.
4.7 Metal spatula
4.8 Disposable Pasteur pipettes and bulbs.
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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 reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Carbon dioxide gas (C02). 99.9 percent purity.
5.4 Oxygen (02). 99.9 percent purity.
5.5 Ethyl acetate (C4H802). Pesticide quality or equivalent.
5.6 1,2,4-Trichlorobenzene (C6H3C13). 99 percent.
5.7 Acetic acid (C2H402), 70% in water. Dilute 7 volumes of acetic acid
with 3 volumes of water.
5.8 Trichlorobenzene solution (C6H3C13), stock (1 jtiL = jug Cl). Prepare
a stock solution by accurately delivering 117 /iL (170 mg) of trichlorobenzene
into a 100-mL volumetric flask and dilute to volume with ethyl acetate.
5.9 Trichlorobenzene solution (C6H3C13), calibration (1 /uL = 100 ng Cl).
Dilute 10 ml of the trichlorobenzene stock solution to 100 ml with ethyl acetate.
5.10 Sodium chloride (NaCl) calibration standard, (1 /zg CV/juL).
Accurately weigh 0.1648 g of sodium chloride into a 100-mL volumetric flask.
Dilute to volume with reagent water.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine.
6.2 All samples must be iced or refrigerated from the time of collection
until analysis.
6.3 All samples should be collected in bottles (at least 25 mL) with
Teflon* septa and be protected from light. If this is not possible, use amber
glass 250-mL bottles fitted with Teflon -lined caps. Foil may be substituted for
Teflon* if the sample is not corrosive. Fill the sample bottle as completely as
possible to minimize headspace until time of analysis. Samples must be preserved
by acidification to pH < 2 with sulfuric acid, stored at 4°C, and protected
against loss of volatiles by eliminating headspace in the container. Samples
9023 - 3 Revision 0
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should be analyzed within 28 days. The container must be washed and muffled at
400°C before use, to minimize contamination.
6.4 If the analysis is to be conducted on suspended solids from a
wastewater sample, isolate the solids by centrifugation, weigh the wet solids,
and analyze immediately. Determine the dry weight of a separate portion of the
wet solids by heating overnight at 110'C.
6.5 All glassware must be dried prior to use according to the protocols
discussed in Sec 3.1.1.
7.0 PROCEDURE
7.1 Calibration
7.1.1 Assemble the solvent injection/pyrolysis/microcoulometric
titration apparatus shown in accordance with the manufacturer's
specifications. Adjust the C02 flow to 300 mL/minute and the 02 flow to
100 mL/minute using the auxiliary flow controllers (bypass the flow
controllers). The pyrolysis furnace should be set at 800 ± 10°C. Attach
the titration cell to the pyrolysis tube outlet and fill with electrolyte
(70% acetic acid).
7.1.2 Turn on the instrument and allow the gas flows and
temperatures to stabilize. When the background current of the titration
cell has stabilized, the instrument is ready for use.
7.1.3 Calibrate the microcoulometric titration system for Cl"
detection by injecting various amounts of the sodium chloride calibration
standards directly into the titration cell and integrating the response
using the POX integration mode. The range of sodium chloride amounts
should cover the range of expected sample concentrations and should always
be less than 80 jug Cl". Over the range 1 - 80 jug Cl" the integrated
response should read within 2% or 0.05 jug (whichever is larger) of the
quantity injected. If this calibration requirement is not met then the
instrument sensitivity parameters should be adjusted according to the
manufacturer's specifications to achieve accurate response.
7.1.4 Check the performance of the entire analytical system by
delivering three 25-juL aliquots of the trichlorobenzene calibrate standard
into the furnace at a rate of 1 //L/second. The mean of these three
analyses should be 2.2 - 2.8 /xg Cl and the percent relative standard
deviation should be 5% or less. If these criteria are not met the system
should be checked as described in the instrument maintenance manual in
order to isolate the problem.
7.1.5 Perform a blank ethyl acetate standard (25-juL) each day. If
the integrated response is greater than 0.1 jug Cl", then the system should
be checked for sources of contamination.
9023 - 4 Revision 0
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7.2 Transfer a 1-gram aliquot of the solid sample to a 10 ml vial using
a metal spatula. Add 1 ml of reagent water and 5 ml of ethyl acetate to the
sample and cap tightly.
7.3 Shake the sample vigorously for thirty seconds and then place the
vial in a sonic bath filled with water to a level of -1 inch, or agitate the
suspension directly using a sonic probe, if available. Sonify the sample for 15
minutes if using a sonic bath or 5 minutes if using a sonic probe.
7.4 Allow the suspension to settle for 10 minutes and then transfer the
upper layer (ethyl acetate) to a 15-mL conical centrifuge tube. Cap the tube and
centrifuge at approximately 1000 x g for five minutes.
7.5 Transfer the ethyl acetate layer to a clean 10 ml vial, cap, and
store refrigerated until analyzed.
7.6 For analysis, withdraw a 5 to 25 /iL aliquot of the ethyl acetate
into a microsyringe having a 4-inch long needle or a boat inlet. Place the
pyrolysis/microcoulometer system into the POX integration mode and immediately
pierce the septum and position the tip of the microsyringe into the furnace.
Deliver the sample at a rate of approximately 1 /iL/second and withdraw the
needle when sample delivery is complete.
7.7 After the 10-minute integration cycle is complete record the
integrated response. If the response exceeds the working range of the
instrument, repeat the analysis after dilution of the extract with reagent grade
ethyl acetate.
7.8 Determine the EOX concentration in the sample as follows:
where: EOX Concentration, ug/g as Cl~ = -^—I x 10(
Ws x VT
Qs = Quantity of EOX as ;ug of Cl" in the aliquot injected.
V, = Volume of aliquot injected in /iL.
VE = Total volume of extract in ml.
Ws = Weight of sample extracted in grams.
7.9 Report results in micrograms per gram. When duplicate and spiked
samples are analyzed, report all data obtained with the sample results.
7.10 For samples processed as part of a set where the spiked sample
recovery falls outside of the historically derived control limits, data for the
affected parameters must be labeled as suspect.
7.11 If the aqueous portion of a water sample, from which the suspended
solids are being analyzed, is expected to contain high levels of organic halide,
9023 - 5 Revision 0
January 1995
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a 1-mL aliquot of the centrifuged sample should be analyzed. The solids data
must then be corrected using the following equation:
where: EOX (corrected) = EOXS - EOXW x -^
WD
EOXS = EOX in wet solids, /ug/g as Cl
EOXW = EOX in water sample, pg/g as Cl
Ws = Wet weight of solids, grams
WD = Dry weight of solids, grams
8.0 QUALITY CONTROL
8.1 Each laboratory that uses this method is required to operate a formal
quality control program. The minimum requirements of this program consist of an
initial demonstration of laboratory capability and the analysis of spiked samples
as a continuing check on performance. The laboratory is required to maintain
performance records to define the quality of data that is generated.
8.2 Before performing any analyses, the analyst must demonstrate the
ability to generate acceptable accuracy and precision with this method.
8.2.1 Select a trichlorobenzene spike concentration representative
of the expected levels in the samples. Using stock standards, prepare a
quality control check sample concentrate in ethyl acetate 100 times more
concentrated than the selected concentration.
8.2.2 Place a minimum of six 1-gram aliquots of an uncontaminated
soil sample in 10 mL vials. Spike four of the samples with 10 /zL of the
check sample, cap the vials, shake vigorously, and allow the spike to
equilibrate with the sample by standing overnight. Analyze the aliquots
according to the procedure beginning in Step 7.2.
8.2.3 Calculate the average percent recovery, (R), and the standard
deviation of the percent recovery (S), for the results. Soil background
corrections must be made before R and S calculations are performed.
8.2.4 Acceptance limits for recovery and precision must be derived
from repeated analyses of the standard discussed in section 8.2.1. Base
the accuracy acceptance criteria on +/- 3 standard deviations from the
mean recovery and the precision acceptance criteria on the relative
standard deviation. If the recovery for a particular parameter does not
fall within the control limits for method performance, the results
reported for that parameter in all samples processed as part of the same
set must be qualified as described in Step 7.10.
8.3 The laboratory must spike and analyze a minimum of 10% of all samples
to monitor continuing laboratory performance.
9023 - 6 Revision 0
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8.4 Each day, the analyst must demonstrate, through the analysis of
uncontaminated soil, that interferences from the analytical system are under
control.
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. An MDL of 10 jitg/g was obtained using
ethyl acetate standards. The MDL actually achieved in a given analysis will vary
depending on instrument sensitivity and matrix effects.
9.2 In a single laboratory, using solid spiked at various levels, the
average recoveries presented in Table 1 were obtained.
10.0 REFERENCES
10.1 "Development and Evaluation of Methods for Total Organic Halide and
Purgeable Organic Halide in Wastewater". EPA-600/4-84-008, PB84-134337 (NTIS).
U.S. Environmental Protection Agency, Environmental Monitoring and Support
Laboratory - Cincinnati, Ohio 45268, January 1984.
9023 - 7 Revision 0
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METHOD 9023
EXTRACTABLE ORGANIC HALIDES (EOX) IN SOLIDS
^
'
7.1.1 - 7.1.2 Assemble
and adjust gas flow
and allow temp, and
gas flow to stabilize.
>
r
7.1.3 - 7.1.5 Calibrate,
check the performance
and run standard on
the instrument.
1
f
7.2 Dissolve 1 g of solid
sample in reagent with
ethyl acetate solution.
7.3 Shake and
sonicate mixture.
7.4 Transfer upper level
to and centrifuge.
7.5 Transfer the ethyl
acetate layer and store
for analysis.
7.6 - 7.7 Analyze sample
and run 10 minute
integration cycle.
7.8 Determine EOX
concentration and report.
Revision 0
January 1995
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METHOD 9057
DETERMINATION OF CHLORIDE FROM HC1/C1, EMISSION SAMPLING
TRAIN (METHODS 0050 AND 0051) BY ANION CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 This method describes the analytical protocol for determination of
hydrogen chloride (HC1, CAS Registry Number 7647-01-0) and chloride (C12 CAS
Registry Number 7782-50-5) in stack gas emission samples collected from hazardous
waste and municipal waste incinerators using the midget impinger HC1/C12 sampling
train (Method 0051) or the isokinetic sampling train (Method 0050).
1.2 The lower detection limit is 0.1 /zg of chloride (CV) per mL of
sample solution. Samples with concentrations which exceed the linear range of
the analytical instrumentation may be diluted.
1.3 This method is recommended for use only by analysts experienced in
the use of ion chromatography and in the interpretation of ion chromatograms.
2.0 SUMMARY OF METHOD
2.1 The stoichiometry of HC1 and HC12 collection in the sampling train
(see Methods 0050 and 0051) is as follows: In the acidified water absorbing
solution, The HC1 gas is solubilized and forms chloride ions (C1-) according to
the following formula:
HC1 + H20 = H30+ + CV
The C12 gas present in the emissions has a very low solubility in acidified water
and passes through to the alkaline absorbing solution where it undergoes
hydrolysis to form a proton (H+), Cl" , and hypochlorous acid (HC10) as shown:
H20 + C12 = H+ +CV + HC10
Non-suppressed or suppressed ion chromatography (1C) is used for analysis of the
CT.
3.0 INTERFERENCES
3.1 Volatile materials which produce chloride ions upon dissolution
during sampling are obvious interferences in the measurement of HC1. One likely
interferant is diatomic chlorine (C12) gas which disproportionates to HC1 and
hypochlorous acid (HOC1) upon dissolution in water. C12 gas exhibits a low
solubility in water, however, and the use of acidic rather than neutral or basic
solutions for collection of hydrogen chloride gas greatly reduces the dissolution
of any chlorine present. Sampling a 400 ppm HC1 gas stream containing 50 ppm C12
with this method does not cause a significant bias. Sampling a 220 ppm HC1 gas
stream containing 180 ppm C12 results in a positive bias of 3.4 percent in the
HC1 measurement. Other interferants have not been encountered.
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3.2 Reducing agents such as S02 may cause a positive bias in the C12
measurement by the following reaction:
HC10 + HS03- = H2S04 + CV
4.0 APPARATUS AND MATERIALS
4.1 Volumetric Flasks. Class A, various sizes, as appropriate.
4.2 Volumetric Pipettes. Class A, assortment, to dilute samples to
calibration range of the 1C.
4.3 Ion Chromatograph. Suppressed or non-suppressed, with a conductivity
detector and electronic integrator operating in the peak area mode. Other
detectors, a strip chart recorder, and peak heights may be used provided the 5
percent repeatability criteria for sample analysis and the linearity criteria for
the calibration curve can be met.
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 Reagent water. All references to water in the method refer to
reagent water as specified by definition in Chapter One.
5.3 Sulfuric acid (0.1 N), H2S04. To prepare 100 ml, slowly add 0.28 ml
of concentrated H2S04 to about 90 ml of water while stirring, and adjust the
final volume to 100 ml using additional water. Shake well to mix the solution.
5.4 Sodium hydroxide (0.1 N), NaOH. To prepare 100 ml, dissolve 0.40 g
of solid NaOH in about 90 ml of water and adjust the final volume to 100 ml using
additional water. Shake well to mix the solution.
5.5 Reagent blank solutions. A separate blank solution of each sampling
train regent used and collected in the field (0.1 N H2S04 and 0.1 N NaOH) should
be prepared for analysis with the field samples. For midget impinger train
sample analysis, dilute 30 ml of each reagent with rinse water collected in the
as a blank to the final volume of the samples. For isokinetic train sample
analysis, dilute 200 ml to the same final volume as the field samples also using
the blank sample of rinse water.
5.6 Sodium chloride, NaCl, stock standard solution. Solutions containing
a nominal certified concentration of 1000 mg/L NaCl are commercially available
as convenient stock solutions from which working standards can be made by
appropriate volumetric dilution. Alternately, concentrated stock solutions may
be produced from reagent grade NaCl that has been dried at 110'C for two or more
hours and then cooled to room temperature in a desiccator immediately before
weighing. Accurately weigh 1.6 to 1.7 g of the dried NaCl to within 0.0001 g,
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dissolve in water, and dilute to 1 liter. The exact Cl" concentration can be
calculated using the equation:
M9 Cr/mL = g of NaCl x 1000 x 35.453/58.44
Refrigerate the stock standard solutions and store no longer than one month.
5.7 Chromatographic eluent. Effective eluent for non-suppressed ion
chromatography using a resin- or silica-based weak ion exchange column are a 4
mM potassium hydrogen phthalate solution, adjusted to a pH of 4.0 using a
saturated sodium borate solution, and a mM 4-hydroxy benzoate solution, adjusted
to a pH of 8.6 using 1 N sodium hydroxide. An effective eluent for suppressed
ion chromatography is a solution containing 3 mM sodium bicarbonate and 2.4 mM
sodium carbonate. Other dilute solutions buffered to a similar pH that contain
no ions interfering with the Chromatographic analysis may be used. If, using
suppressed ion chromatography, the "water dip" resulting from sample injection
is interfering with the chloride peak, use a 2 mM sodium hydroxide/2.4 mM sodium
bicarbonate eluent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING.
6.1 Sample collection using the midget impinger HC1/C12 train or the
isokinetic HC1/C12 train is described in Method 0051 or 0050, respectively.
6.2 Samples should be stored in clearly labeled, tightly sealed
containers between sample recovery and analysis. They may be analyzed up to four
weeks after collection.
7.0 PROCEDURE
7.1 Sample preparation for analysis. Check the liquid level in each
sample, and determine if any sample was lost during shipment. If a noticeable
amount of leakage has occurred, the volume can be determined from the difference
between the initial and final solution levels, and this value can be used to
correct the analytical results. For midget impinger train samples,
quantitatively transfer each sample solution to a 100-mL volumetric flask and
dilute to 100 mL with water. For isokinetic sampling train samples,
quantitatively transfer each sample to a volumetric flask or graduated cylinder
and dilute with water to a final volume appropriate for all samples.
7.2 Calibration of Ion Chromatograph.
7.2.1 The ion Chromatographic conditions will depend on the type of
analytical column used and whether suppressed or non-suppressed ion
chromatography is used. Prior to calibration and sample analysis,
establish a stable baseline. Next, inject a sample of water, and
determine if any CV appears in the chromatogram. If Cl" is present, repeat
the load/injection procedure until no Cl" is present.
7.2.2 To prepare the calibration standards, dilute given amounts
(1.0 mL or greater) of the stock standard solution to convenient volumes,
using 0.1 N H2S04 or 0.1 N NaOH as appropriate. Prepare at least four
standards that are within the linear range of the field samples. Inject
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the calibration standards, starting with the lowest concentration standard
first, both before and after injection of the quality control check
sample, reagent blank, and field samples. This allows compensation for
any instrument drift occurring during sample analysis.
7.2.3 Determine the peak areas, or heights, of the standards and
plot individual values versus CV concentrations in jug/mL. Draw a smooth
curve through the points. Use linear regression to calculate a formula
describing the resulting linear curve.
7.3 Sample analysis. Between injections of the series of calibration
standards, inject, in duplicate, the reagent blanks and the field samples,
including a matrix spike sample. Measure the areas or heights (same as done for
the calibration standards) of the CV peaks. Use the average response to
determine the concentrations of the field samples, matrix spike, and reagent
blanks using the linear calibration curve, the results for a reagent blank
should not exceed 10 percent of the corresponding value for a field sample.
7.4 Calculations. Retain at least one extra decimal figure beyond those
contained in the available data in intermediate calculations, and round off only
the final answer appropriately.
7.4.1 Total /j,g HC1 per sample. Calculate as described below:
mHci = (S'B) x vs x 36.46/35.453 (1)
where:
mHd = Mass of HC1 in sample, y^g,
S = Analysis of sample, jug Cl'/mL,
B = Analysis of reagent blank, /ug CV/mL,
Vs = Volume of filtered and diluted sample, ml
36.46 = Molecular weight of HC1, /ig/^g-mole,
and
35.45 = Atomic weight of CV, jug//Ltg-mole.
7.4.2 Total nq C12 per sample. Calculate as described below:
mcl2 = (S-B) x V2 x 70.91/35.45 (2)
where:
mcl2 = Mass of C12 in sample, /zg
70.91 = Molecular weight of C12 /ig/jug-mole,
and
35.45 = Atomic weight of CV, /Ltg//ig-mole.
7.4.3 Concentration of HC1 in the flue gas: Calculate as described
below:
C = K x m/V
m(std)
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where:
C = Concentration of HC1 or C12, dry basis, mg/dscm,
K = 10"3 mg/jug,
m = Mass of HC1 or C12 in sample, ng
and
Vm(stdi = Dry 9as volume measured by the dry gas meter,
corrected to standard conditions, dscm (from Method 0050 or
Method 0051).
8.0 QUALITY CONTROL
8.1 At the present time, a validated audit material does not exist for
the method. However, it is strongly recommended that a quality control check
sample and a matrix spike sample be used.
8.1.1 Quality control check sample. Chloride solutions of reliably
known concentrations are available for purchase from the National
Institute of Science and Technology (SRM 3182). The QC check sample
should be prepared in the appropriate absorbing reagent at a concentration
approximately equal to the mid-range calibration standard, The quality
control check sample should be injected, in duplicate, immediately after
the calibration standards have been injected for the first time. The CV
value obtained for the check sample using the final calibration curve
should be within 10 percent of the known value for the check sample.
8.1.2 Matrix spike sample. A portion of at least one field sample
should be used to prepare a matrix spike sample. Spike the sample aliquot
in the range of the expected concentration. Analyze the matrix spike
sample in duplicate along with the field samples. Based on the matrix
spike results, determine the recovery for the spiked material. This
should be within 10 percent of the known spike value.
8.2 Refer to Chapter One for additional quality control criteria.
9.0 METHOD PERFORMANCE
9.1 The lower detection limit of the analytical method is 0.1 jug of Cl"
per mL of sample solution. Samples with concentrations which exceed the linear
range of the 1C may be diluted.
9.2 The precision and bias of for analysis of HC1 using this analytical
protocol have been measured in combination with the midget impinger HC1/C12 train
(Method 0051) for sample collection. The within-laboratory relative standard
deviation is 6.2 percent and 3.2 percent at HC1 concentrations of 3.9 and 15.3
ppm, respectively. The method does not exhibit any bias for HCl when sampling
at C12 concentrations less than 50 ppm.
10.0 REFERENCES
1. Steinsberger, S.C. and J. H. Margeson, "Laboratory and Field Evaluation of
a Methodology for Determination of Hydrogen Chloride Emissions from
Municipal and Hazardous Waste Incinerators," U.S. Environmental Protection
9057 - 5 Revision 0
January 1995
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Agency, Office of Research and Development, Report No. EPA 600/3-89/064,
NTIS PB89 220586-AS.
2. State of California, Air Resources Board, Method 421, "Determination of
Hydrochloric Acid Emissions from Stationary Sources" March 18, 1987.
3. Entropy Environmentalists, Inc., "Laboratory Evaluation of a Sampling and
Analysis Method for Hydrogen Chloride emissions from Stationary Sources:
Interim Report," EPA Contract No. 68-02-4442, Research Triangle Park,
North Carolina, January 22, 1988.
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METHOD 9057
DETERMINATION OF CHLORIDE FROM HC1/C1, EMISSION SAMPLING
TRAIN (METHODS 0050 AND 0051) BY ANION CHROMATOGRAPHY
7.2.1 Determine
1C conditions.
I
7.2.2 Prepare
calibration
standards.
7.2.3 Dtermine
peak areas of
standards.
7.3 Conduct sample
analysis with QC
and blanks.
I
7.4 Perform
calculations.
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Revision 0
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METHOD 9210
POTENTIOMETRIC DETERMINATION OF NITRATE
IN AQUEOUS SAMPLES WITH ION-SELECTIVE ELECTRODE
1.0 SCOPE AND APPLICATION
1.1 This method can be used for measuring total solubilized nitrate in
drinking waters, natural surface waters, groundwaters, domestic and industrial
wastewaters, and in soil extracts (ASTM methods D4646-87, D5233-92 or D3987-
85).
NOTE: This method is for the analysis of simple nitrate ion rather
than total nitrate, as analysis using the ion-selective electrode
is not preceded by a distillation step.
1.2 The method detection limit is 2.0 mg/L. Nitrate concentrations
from 0.2 to 1,000 mg/L may be measured. However, using a linear calibration,
results less than 2 mg/L may be biased up to approximately 420 percent high;
results greater than 400 mg/L may be biased up to approximately 50 percent
low.
1.3 ISEs must be used carefully, and results must be interpreted
cautiously, since an ISE may be affected by numerous analytical interferences
which may either increase or decrease the apparent analyte concentration, or
which may damage the ISE. Effects of most interferences can be minimized or
eliminated by adding appropriate chemical reagents to the sample. Obtaining
the most accurate results, therefore, requires some knowledge of the sample
composition.
NOTE: ISE manufacturers usually include a list of interferences in
the instruction manual accompanying an ISE, along with recommended
methods for minimizing or eliminating effects of these
interferences.
2.0 SUMMARY OF METHOD
2.1 Total solubilized nitrate is determined potentiometrically using a
nitrate ion-selective electrode (ISE) in conjunction with a double-junction
reference electrode and a pH meter with an expanded millivolt scale or an ISE
meter capable of being calibrated directly in terms of nitrate concentration.
2.2 Standards and samples are mixed 50:1 with an ionic strength
adjustment solution (ISA). Calibration is performed by analyzing a series of
standards and plotting mV vs. nitrate-nitrogen concentration on semilog paper
or by calibrating the ion meter directly in terms of nitrate concentration.
3.0 INTERFERENCES
3.1 The nitrate electrode responds to numerous interfering anions.
Most of the interferants, however, can be rendered harmless by adding suitable
reagents. Cyanide, bisulfide, bicarbonate, carbonate, and phosphate are
removed by adjusting the solution pH to 4 with boric acid. Chloride, bromide,
9210 - 1 Revision 0
January 1995
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and iodide are removed by adding silver sulfate solution. Nitrite is also an
interferant, as shown in Table 1; nitrite is removed by adding sulfamic acid.
The amounts of silver sulfate and sulfamic acid needed will vary based on the
concentrations of interferants. As a general guide, 1 ml of silver sulfate
will eliminate chloride interference in a 50 ml sample containing 35 mg/L Cl";
1 mL of sulfamic acid solution will eliminate nitrite interference in a 50 ml
sample containing 95 mg/L N02".
3.2 Temperature changes affect electrode potentials. Using an ISE
calibrated at 22°C, a 20.0 mg/L nitrate solution was measured as 20.6 mg/L at
22 "C and 12.9 mg/L at 32°C (see Ref. 4). Therefore, standards and samples
must be equilibrated at the same temperature (+ 1°C).
3.3 The user should be aware of the potential of interferences from
colloidal substances and that, if necessary, the samples may be filtered.
4.0 APPARATUS AND MATERIALS
4.1 pH/mV meter capable of reading to 0.1 mV or an ISE meter.
4.2 Nitrate ISE (Orion 9307 or equivalent) and double-junction
reference electrode (Orion 9002 or equivalent).
4.3 Thermally isolated magnetic stirrer, Teflon®-coated stir bar, and
stopwatch.
4.4 Volumetric flask, 100 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 reagent is of sufficiently
high purity to permit its use without lessening the accuracy of the
determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 ISA adjuster solution (2M, (NH4)2S04): Dissolve 26.4 g of
ammonium sulfate in reagent water to make 100 mL of solution.
5.4 Boric acid (1M, H3B03): Dissolve 6.2 g of boric acid in reagent
water to make 100 mL of preservative solution (for numerous anions and
bacteria).
5.5 Silver sulfate (0.05 M, Ag2S04) to remove interferences noted in
Step 3.1. A saturated silver sulfate solution contains approximately 5.5 g/L
of solubilized silver.
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5.6 Sulfamic acid (0.1 M, HOS02NH2) to remove nitrite from sample, as
noted in Step 3.1.
5.7 Nitrate calibration stock solution (1,000 mg/L, N03"): Dissolve
0.1631 g of potassium nitrate (dried two hours at 110'C and stored in a
desiccator) in reagent water, add 1.00 ml of preservative solution, and dilute
to 100 ml in a volumetric flask. Store in a clean bottle.
5.8 Nitrate calibration standards: Prepare a series of calibration
standards by diluting the 1,000 mg/L nitrate standard. A suitable series is
given in the table below.
ml of 1,000 mg/L Concentration when Diluted
N03- Solution to 50.0 mL (mg/L N03'-N)
0.0500 1.00
0.150 3.00
0.500 10.0
1.50 30.0
5.00 100.0
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 In most environmental samples, nitrate is not affected by
complexation, precipitation, inorganic oxidation-reduction reactions, and
protonation. In the presence of a reducing agent (e.g., organic matter),
however, bacteria will utilize nitrate as an oxidant, causing a slow decrease
in the nitrate concentration. This potential interference can be obviated by
using a preservative. Therefore, samples must be preserved by adding 1 mL of
1M boric acid solution per 100 mL of sample.
6.3 Samples should be stored at 4° C and must be analyzed within three
(3) days of collection.
7.0 PROCEDURE
7.1 Calibration
7.1.1 When using a nitrate ISE and a separate double-junction
reference electrode, ensure that reference electrode inner and outer
chambers are filled with solutions recommended by the manufacturer.
Equilibrate the electrodes for at least one hour in a 100 mg/L nitrate
standard before use.
7.1.2 Calibrate the nitrate ISE using standards that narrowly
bracket the expected sample concentration. If the sample concentration
is unknown, calibrate with 3.00 mg/L and 30.0 mg/L nitrate standards.
Add 50.0 mL of standard, 0.50 mL of preservative solution, and 1.00 mL
of ISA to a 100-mL beaker. Add a Teflon®-coated magnetic stir bar,
9210 - 3 Revision 0
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place the beaker on a magnetic stir plate, and stir at slow speed (no
visible vortex). Immerse the electrode tips to just above the rotating
stir bar. If using an ISE meter, calibrate the meter in terms of
nitrate concentration following the manufacturer's instructions. If
using a pH/mV meter, record the meter reading (mV) as soon as the
reading is stable, but in no case should the time exceed five minutes
after immersing the electrode tips. Prepare a calibration curve by
plotting measured potential (mV) as a function of the logarithm of
nitrate concentration. The slope must be 54-60 mV per decade of nitrate
concentration. If the slope is not acceptable, the ISE may not be
working properly. For corrective action, consult the ISE operating
manual.
7.2 Allow samples and standards to equilibrate to room temperature.
7.3 Prior to and between analyses, rinse the electrodes thoroughly
with reagent water and gently shake off excess water. Low-level measurements
are faster if the electrode tips are first immersed five minutes in reagent
water.
7.4 Add 50.0 ml of sample and 1.00 ml of ISA to a 100-mL beaker. Add
a Teflon®-coated magnetic stir bar. Place the beaker on a magnetic stir plate
and stir at a slow speed (no visible vortex). Immerse the electrode tips to
just above the rotating stir bar. Record the meter reading (mV or
concentration) as soon as the reading is stable, but in no case should the
time exceed five minutes after immersing the electrode tips. If reading mV,
determine nitrate-nitrogen concentration from the calibration curve.
7.5 When analyses have been completed, rinse the electrodes thoroughly
and store them in a 100 mg/L nitrate standard solution. If the electrodes
will not be used more than one day, drain the reference electrode internal
filling solutions, rinse with reagent water, and store dry.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Initial Calibration Verification standard (ICV): After performing
the calibration step (7.1), verify calibration by analyzing an ICV. The ICV
contains a known nitrate concentration at the mid-range of the calibration
standards and is from an independent source. ICV recovery must be 90-110
percent. If not, the source of error must be found and corrected. An
acceptable ICV must be analyzed prior to sample analysis. The ICV also serves
as a laboratory control sample.
8.3 Continuing Calibration Verification standard (CCV): After every
10 samples, and after the final sample, a CCV must be analyzed. The CCV
contains a known nitrate concentration at mid-calibration range. CCV recovery
must be 90-110 percent. If not, the error source must be found and corrected.
If ISE calibration has changed, all samples analyzed since the last acceptable
CCV must be re-analyzed.
8.4 Reagent blank: After the ICV and after every CCV, a reagent blank
must be analyzed. A reagent blank is a 1 percent solution of preservative
9210 - 4 Revision 0
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solution in reagent water, mixed 50:1 with ISA. The indicated reagent blank
concentration must be less than 1 mg/L nitrate. If not, the contamination
source must be found and corrected. All samples analyzed since the last
acceptable reagent blank must be re-analyzed.
8.5 Matrix spike: Follow the matrix spike protocols presented in
Chapter One. The spike concentration must be 10 times the detection limit and
the volume added must be negligible (less than or equal to one-thousandth the
sample aliquot volume). Spike recovery must be 75-125 percent. If not,
samples must be analyzed by the method of standard additions.
9.0 METHOD PERFORMANCE
9.1 In a single-laboratory evaluation, a series of standards with
known nitrate concentrations was analyzed with a nitrate ISE. Measurements
were obtained over three consecutive days using an Orion 9307 nitrate ISE and
an Orion 9002 double-junction reference electrode connected to an Orion 940
ISE meter. A two-point calibration (5.00 and 50.0 mg/L nitrate) was performed
prior to analysis. The results are listed in Table 2.
9.2 In the same study, three groundwater samples were spiked with
nitrate at four different concentrations and measured with the nitrate ISE.
(The groundwater samples initially contained <0.1-2.3 mg/L nitrate.) Each
spiked sample was analyzed at each concentration, and the mean recoveries and
RSDs are given in Table 3.
9.3 A 50 g portion of soil, which initially contained 0.7 mg/kg
nitrate, was spiked with 25.0 mg/kg nitrate to obtain an anion concentration
in a single extract volume within the linear range of the ISE. The extract
was then analyzed for nitrate using this ISE method, and 89% of the soil spike
was recovered.
10.0 REFERENCES
1. Franson, Mary Ann H., Ed. Standard Methods for the Examination of
Water and Wastewater, 18th Edition. American Public Health Association,
Washington, DC, 1992.
2. Model 93-07 Nitrate Electrode Instruction Manual. Orion Research,
Inc., Boston, MA, 1986.
3. Miller, E.L., Waltman, D.W., and Hillman, D.C. Single-Laboratory
Evaluation of Fluoride, Chloride, Bromide, Cyanide, and Nitrate Ion-Selective
Electrodes for Use in SW-846 Methods. Lockheed Engineering and Sciences
Company for Environmental Monitoring Systems Laboratory, U.S. EPA. September
1990. EPA/600/X-90/221.
9210 - 5 Revision 0
January 1995
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Table 1. Nitrate ISE Interferences
Analyte
Concentration
(mg/L)
25.0
25.0
25.0
25.0
Interference
None
0.01 M H2S04
100 mg/L N02'
100 mg/L N02' +
500 mg/L HOS02NH2
Measured
Concentration
(mg/L)
26
24.5
46
26
RSD
(%)
6.2
5.9
9.1
6.3
Table 2. Results from a Single-Laboratory Accuracy
Evaluation of a Nitrate ISE
Nitrate
Concentration
(mg/L)
0.100
0.200
0.500
1.00
2.00
5.00
10.0
20.0
50.0
100
200
400
1,000
Nitrate
Detected
(mg/L)
1.01
1.04
1.23
1.71
2.45
5.0
11.0
18.9
50
96
164
310
480
Nitrate
Recovery
(percent)
1,010
520
246
171
123
100
110
95
100
96
82
77
48
Rel. Std.
Deviation
(percent)
53
17
8
2
7
0
8
14
1
13
3
8
17
9210 - 6
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Table 3. Mean Spike Recoveries of Nitrate in Three Groundwater Samples
Analyte Spike Spike Rel. Std.
Concentration Recovery Deviation
(mg/L) (percent) (percent)
2.00 113 10.7
3.00 106 7.6
5.00 98 1.2
10.0 89 2.7
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METHOD 9210
POTENTIOMETRIC DETERMINATION OF NITRATE
IN AQUEOUS SAMPLES WITH ION-SELECTIVE ELECTRODE
i
7.1.1 - 7.1.2
Calibrate Nitrate
ISE.
1
r
7.2 Allow standards
to equilibrate to
room temperature.
^
r
7.3 Rinse
eletrodes.
>
r
7.4 Measure
concentration using
electrode meter
and calculate
concentration.
^
r
7.5 Drain
reference electrode
and clean.
3
r
9210 - 8
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METHOD 9211
POTENTIOMETRIC DETERMINATION OF BROMIDE
IN AQUEOUS SAMPLES WITH ION-SELECTIVE ELECTRODE
1.0 SCOPE AND APPLICATION
1.1 This method can be used for measuring total solubilized bromide in
drinking waters, natural surface waters, groundwaters, domestic and industrial
wastewaters, and in soil extracts (ASTM methods D4646-87, D5233-92 or D3987-85).
NOTE: This method is for the analysis of simple bromide ion rather
than total bromide, as analysis using the ion-selective electrode is
not preceded by a distillation step.
1.2 The method detection limit is 0.2 mg/L. Bromide concentrations from
0.1 to 1,000 mg/L may be measured. However, when using a linear calibration,
results less than 0.2 mg/L may be biased up to approximately 40 percent high.
1.3 ISEs must be used carefully, and results must be interpreted
cautiously, since an ISE may be affected by numerous analytical interferences
which may either increase or decrease the apparent analyte concentration, or
which may damage the ISE. Effects of most interferences can be minimized or
eliminated by adding appropriate chemical reagents to the sample. Obtaining the
most accurate results, therefore, requires some knowledge of the sample
composition.
NOTE: ISE manufacturers usually include a list of interferences in
the instruction manual accompanying an ISE, along with recommended
methods for minimizing or eliminating effects of these
interferences.
2.0 SUMMARY OF METHOD
2.1 Total solubilized bromide is determined potentiometrically using a
bromide ion-selective electrode (ISE) in conjunction with a double-junction
reference electrode and a pH meter with an expanded millivolt scale or an ISE
meter capable of being calibrated directly in terms of bromide concentration.
2.2 Standards and samples are mixed 50:1 with an ionic strength
adjustment solution (ISA). Calibration is performed by analyzing a series of
standards and plotting mV vs. bromide concentration on semilog paper or by
calibrating the ion meter directly in terms of bromide concentration.
3.0 INTERFERENCES
3.1 Some polyvalent cations (e.g., Fe*3 and Al+3) at high concentrations
(> 300 mg/L) interfere by forming complexes with bromide which are not measured
by the bromide ISE. However, in dilute aqueous solutions, aluminum and iron(III)
do not form complexes with bromide ions and dilute sulfuric acid has no effect
on the bromide concentration because silver sulfate is soluble and because
hydrogen bromide is a strong acid in water. Chloride is not an interference at
9211 - 1 Revision 0
January 1995
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10 times the mass concentration of bromide since the solubility product of silver
chloride is about 400 times that of silver bromide. (See Table 1.) Bromide
concentration in the presence of very high concentrations of polyvalent cations
can be measured by treating the sample with an equal volume of EDTA or by the
method of standard additions. Solutions containing much Fe+3 will be colored
brownish-red at pH values of 1 or more; in such cases, phosphoric acid or sodium
phosphate monobasic can be added until the solution is colorless (assuming the
only coloring reagent is Fe+3), at which point the iron interference will have
been removed and bromide can be accurately determined by the ISE.
3.2 Sulfide, cyanide, and ammonia interfere with the determination by
reacting directly with the ISE. These interferences can be removed by acidifying
the sample to a pH of 4 with dilute sulfuric acid.
3.3 Temperature changes affect electrode potentials. Using an ISE
calibrated at 22°C, a 20.0 mg/L chloride solution was measured as 20.2 mg/L at
22°C and 14.2 mg/L at 32° (see Ref. 4). Therefore, standards and samples must
be equilibrated at the same temperature (+ 1°C).
3.4 The user should be aware of the potential of intereferences from
colloidal substances and that, if necessary, the samples may be filtered.
4.0 APPARATUS AND MATERIALS
4.1 pH/mV meter capable of reading to 0.1 mV or an ISE meter.
4.2 Bromide ISE (Orion 9435 or equivalent) and double-junction reference
electrode (Orion 9002 or equivalent).
4.3 Thermally isolated magnetic stirrer, Teflon®-coated stir bar, and
stopwatch.
4.4 Volumetric flask, 100 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 reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 ISA solution (5M NaN03): Dissolve 42.5 g of sodium nitrate in
sufficient reagent water to make 100 mL of solution. Store in a clean glass or
plastic container.
5.4 Ethylenediaminetetraacetate (EDTA), disodium salt (2% C10H12N208Na2).
Use as directed in Step 3.1.
9211 - 2 Revision 0
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5.5 Phosphoric acid (H3P04). Use as directed in Step 3.1.
5.6 Sodium phosphate monobasic (NaH2P04 • H20). Use as directed in Step
3.1.
5.7 Dilute sulfuric acid (0.01 M, H2S04): Use as directed in Step 3.2.
5.8 Bromide calibration stock solution (1,000 mg/L Br"): Dissolve 0.1489
g of potassium bromide (dried two hours at 110'C and stored in a desiccator) in
reagent water and dilute to 100 ml in a volumetric flask. Store in a clean
bottle.
5.9 Bromide standard solution (100 mg/L Br"): Dilute 10.0 ml of
1,000 mg/L bromide calibration stock solution to 100 mL with reagent water in a
volumetric flask.
5.10 Bromide calibration standards: Prepare a series of calibration
standards by diluting the 100 mg/L bromide standard. A suitable series is given
in the table below.
mL of 100 mg/L Concentration when Diluted
Br" Solution to 50.0 mL (mg/L Br")
0.150 0.300
0.500 1.00
1.50 3.00
5.00 10.0
15.0 30.0
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 Samples should be stored at 4° C prior to analysis.
7.0 PROCEDURE
7.1 Calibration
7.1.1 When using a bromide ISE and a separate double-junction
reference electrode, ensure that reference electrode inner and outer
chambers are filled with the solutions recommended by the manufacturer.
Equilibrate the electrodes for at least one hour in a 3.00 mg/L bromide
standard before use.
7.1.2 Calibrate the bromide ISE using standards that narrowly
bracket the expected sample concentration. If the sample concentration is
unknown, calibrate with 1.00 mg/L and 10.0 mg/L bromide standards. Add
50.0 mL of standard and 1.00 mL of ISA to a 100-mL beaker. Add a Teflon®-
coated magnetic stir bar, place the beaker on a magnetic stir plate, and
9211 - 3 Revision 0
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stir at slow speed (no visible vortex). Immerse the electrode tips to
just above the rotating stir bar. If using an ISE meter, calibrate the
meter in terms of bromide concentration following the manufacturer's
instructions. If using a pH/mV meter, record the meter reading (mV) as
soon as the reading is stable, but in no case should the time exceed five
minutes after immersing the electrode tips. Prepare a calibration curve
by plotting measured potential (mV) as a function of the logarithm of
bromide concentration. The slope must be 54-60 mV per decade of bromide
concentration. If the slope is not acceptable, the ISE may not be working
properly. For corrective action, consult the ISE operating manual.
7.2 Allow samples and standards to equilibrate to room temperature.
7.3 Prior to and between analyses, rinse the electrodes thoroughly with
reagent water and gently shake off excess water. Low-level measurements are
faster if the electrode tips are first immersed for five minutes in reagent
water.
7.4 Add 50.0 ml of sample and 1.00 ml of ISA to a 100 mL beaker. Add a
Teflon®-coated magnetic stir bar. Place the beaker on a magnetic stir plate and
stir at a slow speed (no visible vortex). Immerse the electrode tips to just
above the rotating stir bar. Record the meter reading (mV or concentration) as
soon as the reading is stable, but in no case should the time exceed five minutes
after immersing the electrode tips. If reading mV, determine bromide
concentration from the calibration curve.
7.5 When analyses have been completed, rinse the electrodes thoroughly
and store them in a 3.00 mg/L bromide standard solution. If the electrodes will
not be used more than one day, drain the reference electrode internal filling
solutions, rinse with reagent water, and store dry.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Initial Calibration Verification standard (ICV): After performing
the calibration step (7.1), verify calibration by analyzing an ICV. The ICV
contains a known bromide concentration at the mid-range of the calibration
standards and is from an independent source. ICV recovery must be 90-110
percent. If not, the error source must be found and corrected. An acceptable
ICV must be analyzed prior to sample analysis. The ICV also serves as a
laboratory control sample.
8.3 Continuing Calibration Verification standard (CCV): After every 10
samples, and after the final sample, a CCV must be analyzed. The CCV contains
a known bromide concentration at mid-calibration range. CCV recovery must be 90-
110 percent. If not, the error source must be found and corrected. If ISE
calibration has changed, all samples analyzed since the last acceptable CCV must
be re-analyzed.
8.4 Reagent blank: After the ICV and after every CCV, a reagent blank
must be analyzed. A reagent blank is reagent water mixed 50:1 with ISA. The
indicated reagent blank concentration must be less than 0.3 mg/L bromide. If
9211 - 4 Revision 0
January 1995
-------
not, the contamination source must be found and corrected. All samples analyzed
since the last acceptable reagent blank must be re-analyzed.
8.5 Matrix spike: Follow the matrix spike protocols presented in Chapter
One. The spike concentration must be 10 times the detection limit and the volume
added must be negligible (less than or equal to one-thousandth the sample aliquot
volume). Spike recovery must be 75-125 percent. If not, samples must be
analyzed by the method of standard additions.
9.0 METHOD PERFORMANCE
9.1 In a single-laboratory evaluation, a series of standards with known
bromide concentrations was analyzed with a bromide ISE. Measurements were
conducted over three consecutive days using an Orion 9435 bromide ISE and an
Orion 9002 double-junction reference electrode connected to an Orion 940 ISE
meter. A two-point calibration (4.00 and 40.0 mg/L bromide) was performed prior
to analysis. Results are listed in Table 2.
9.2 In the same study, four groundwater samples were spiked with bromide
at four different concentrations and were measured with the bromide ISE. (The
groundwater samples initially contained <0.1-0.2 mg/L bromide.) Each spiked
sample was analyzed at each concentration, and the mean recoveries and RSDs are
given in Table 3.
9.3 A 50 g portion of soil, which initially contained <0.1 mg/kg bromide,
was spiked with 5.00 mg/kg bromide to obtain an anion concentration in a single
extract volume within the linear range of the ISE. The extract was then analyzed
for bromide using this ISE method, and 92% of the soil spike was recovered.
10.0 REFERENCES
1. Franson, Mary Ann H., Ed. Standard Methods for the Examination of
Water and Wastewater, 18th Edition. American Public Health Association,
Washington, DC, 1992.
2. Model 94-35 Bromide Electrode Instruction Manual. Orion Research,
Inc., Boston, MA, 1986.
3. Miller, E.L., Waltman, D.W., and Hillman, D.C. Single-Laboratory
Evaluation of Fluoride, Chloride, Bromide, Cyanide, and Nitrate Ion-Selective
Electrodes for Use in SW-846 Methods. Lockheed Engineering and Sciences Company
for Environmental Monitoring Systems Laboratory, U.S. EPA. September 1990.
EPA/600/X-90/221.
4. Cotton, F. Albert, and Geoffrey Wilkinson; Advanced Inorganic
Chemistry, 2nd Edition; Interscience Publishers, New York, NY; 1966.
5. Weast, Robert C., Ed.; CRC Handbook of Chemistry and Physics, 58th
Edition; CRC Press, Inc., Cleveland, Ohio; 1977.
6. Kolthoff, I.M., E.B. Sandell, E.J. Meehan, and Stanley Bruckenstein;
Quantitative Chemical Analysis, 4th Edition; The MacMillan Company, New York, NY;
1969.
9211 - 5 Revision 0
January 1995
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Table 1. Bromide ISE Interferences
Analyte
Concentration
(mg/L)
20.0
200
20.0
200
20.0
200
200
20.0
Interference
None
None
300 mg/L Fe+3
300 mg/L Fe+3
300 mg/L Al+3
300 mg/L Al+3
0.01 M H2S04
200 mg/L CV
Measured
Concentration
(mg/L)
20.0
200
19.6
196
20.0
196
201
20.1
RSD
(%)
1.3
1.3
2.0
0.8
3.2
2.0
1.5
1.9
Table 2. Results from a Single-Laboratory Accuracy Evaluation
of a Bromide ISE.
Bromide
Concentration
(mg/L)
0.100
0.200
0.400
1.00
2.00
4.00
10.0
20.0
40.0
100.
200.
400.
1,000.
Bromide
Detected
(mg/L)
0.141
0.217
0.40
1.00
2.02
4.0
10.0
20.3
40.
104.
203.
400.
990.
Bromide
Recovery
(percent)
141
109
100
100
101
100
100
102
100
104
102
100
99
Rel. Std.
Deviation
(percent)
17
6
3
1
2
3
3
8
2
4
6
5
6
9211 - 6
Revision 0
January 1995
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Table 3. Mean Spike Recoveries of Bromide in 4 Groundwater Samples
Analyte Spike Spike Rel. Std.
Concentration Recovery Deviation
(mg/L) (percent) (percent)
0.500 96 12.2
1.00 94 1.2
3.00 101 2.3
5.00 96 1.0
9211 - 7 Revision 0
January 1995
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METHOD 9211
POTENTIOMETRIC DETERMINATION OF BROMIDE
IN AQUEOUS SAMPLES WITH ION-SELECTIVE ELECTRODE
^
r
7.1.1 - 7.1.2
Calibrate Bromide
ISE.
^
r
7.2 Allow standards
to equilibrate to
room temperature.
^
r
7.3 Rinse
eletrodes.
>
r
7.4 Measure
concentration using
electrode meter
and calculate
concentration.
•^
r
7.5 Drain
reference electrode
and clean.
i
r
9211 - 8
Revision 0
January 1995
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METHOD 9212
POTENTIOMETRIC DETERMINATION OF CHLORIDE
IN AQUEOUS SAMPLES WITH ION-SELECTIVE ELECTRODE
1.0 SCOPE AND APPLICATION
1.1 This method can be used for measuring total solubilized chloride in
drinking waters, natural surface waters, groundwaters, domestic and industrial
wastewaters, and in soil extracts (ASTM methods D4646-87, D5233-92 or D3987-85).
NOTE: This method is for the analysis of simple chloride ion rather
than total chloride, as analysis using the ion-selective electrode
is not preceded by a distillation step.
1.2 The method detection limit is 2.0 mg/L. Chloride concentrations from
0.4 to 1,000 mg/L may be measured. However, when using a linear calibration,
results less than 2.0 mg/L may be biased up to approximately 60 percent high.
1.3 ISEs must be used carefully and results must be interpreted
cautiously, since an ISE may be affected by numerous analytical interferences
which may either increase or decrease the apparent analyte concentration, or
which may damage the ISE. Effects of most interferences can be minimized or
eliminated by adding appropriate chemical reagents to the sample. Obtaining the
most accurate results, therefore, requires some knowledge of the sample
composition.
NOTE: ISE manufacturers usually include a list of interferences in
the instruction manual accompanying an ISE, along with recommended
methods for minimizing or eliminating effects of these
interferences.
1.4 The chloride ISE should not be used in solutions containing high
concentrations of bromide or iodide, cyanide, or sulfide since these ions form
silver salts less soluble than silver chloride. The chloride ISE will also give
erroneous readings and will be damaged when used in solutions containing free
ammonia, since such solutions dissolve silver chloride.
2.0 SUMMARY OF METHOD
2.1 Total solubilized chloride is determined potentiometrically using a
chloride ion-selective electrode (ISE) in conjunction with a double-junction
reference electrode, or a chloride combination ISE, and a pH meter with an
expanded millivolt scale or an ISE meter capable of being calibrated directly in
terms of chloride concentration.
2.2 Standards and samples are mixed 50:1 with an ionic strength
adjustment solution (ISA). Calibration is performed by analyzing a series of
standards and plotting mV vs. chloride concentration on semi log paper or by
calibrating the ion meter directly in terms of chloride concentration.
9212 - 1 Revision 0
January 1995
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3.0 INTERFERENCES
3.1 Polyvalent cations (e.g., Fe+3 and Al+3) interfere by forming
complexes with chloride which are not measured by the chloride ISE. (See Table
1.) Aluminum and iron(III) ions (at concentrations of 300 mg/L and higher) form
complexes with chloride ions having approximately equal stability. (A sample
with high solubilized Fe+3 and Al concentrations will be distinctly colored
yellow-brown.) Dilute sulfuric acid has no effect on the chloride concentration
because silver sulfate is soluble, and because hydrogen chloride is a strong acid
(completely ionized) in water. Chloride concentration in the presence of high
concentrations of polyvalent cations can be measured by treating the sample with
an equal volume of EDTA or by the method of standard additions.
3.2 Bromide, sulfide, cyanide, and ammonia interfere with the
determination by reacting directly with and damaging the ISE. Sulfide, cyanide,
and ammonia can be removed by acidifying the sample to a pH of 4 with dilute
sulfuric acid. Bromide and iodide can be removed by treating the acidified
sample with potassium bromate, which converts the ions to bromine and iodate.
3.3 Temperature changes affect electrode potentials. Using an ISE
calibrated at 22°C, a 40.0 mg/L chloride solution was measured as 40.0 mg/L at
22°C and 24.8 mg/L at 32° (see Ref. 4). Therefore, standards and samples must
be equilibrated at the same temperature (+ 1°C).
3.4 The user should be aware of the potential of intereferences from
colloidal substances and that, if necessary, the samples may be filtered.
4.0 APPARATUS AND MATERIALS
4.1 pH/mV meter capable of reading to 0.1 mV or an ISE meter.
4.2 Combination chloride ISE (Orion 9617 or equivalent), or separate
chloride ISE (Orion 9417 or equivalent) and double-junction reference electrode
(Orion 9002 or equivalent).
4.3 Thermally isolated magnetic stirrer, Teflon®-coated stir bar, and
stopwatch.
4.4 Volumetric flask, 100 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 reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
9212 - 2 Revision 0
January 1995
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5.3 ISA solution (5M NaN03): Dissolve 42.5 g of sodium nitrate in
sufficient reagent water to make 100 ml of solution. Store in a clean glass or
plastic container.
5.4 Ethylenediaminetetraacetate (EDTA), disodium salt (2% C10H12N208Na2).
Use as directed in Step 3.1.
5.5 Dilute sulfuric acid (0.01 M H2S04): use as directed in Step 3.2.
5.6 Potassium bromate (1%, KBr03): use as directed in Step 3.2.
5.7 Chloride calibration stock solution (1,000 mg/L CV): Dissolve
0.1649 g of sodium chloride (dried two hours at HO'C and stored in a desiccator)
in reagent water and dilute to 100 ml in a volumetric flask. Store in a clean
bottle.
5.8 Chloride calibration standards: Prepare a series of calibration
standards by diluting the 1,000 mg/L chloride standard. A suitable series is
given in the table below.
ml of 1,000 mg/L Concentration when Diluted
CV Solution to 50.0 mL (mg/L CV)
0.050 1.00
0.150 3.00
0.50 10.0
1.50 30.0
5.0 100.0
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 Samples should be stored at 4°C. There are no other special sample
handling or preservation requirements.
7.0 PROCEDURE
7.1 Calibration
7.1.1 If using a chloride combination ISE, ensure that the ISE is
filled with the solution recommended by the manufacturer. Change the
solution if the ISE has not been used for a week. If using a chloride ISE
and a separate double-junction reference electrode, ensure that reference
electrode inner and outer chambers are filled with solutions recommended
by the manufacturer. In either case, equilibrate the electrode(s) for at
least one hour in a 30.0 mg/L chloride standard before use.
7.1.2 Calibrate the chloride ISE using standards that narrowly
bracket the expected sample concentration. If the sample concentration is
9212 - 3 Revision 0
January 1995
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unknown, calibrate with 10.0 mg/L and 100 mg/L chloride standards. Add
50.0 mL of standard and 1.00 mL of ISA to a 100 ml beaker. Add a Teflon®-
coated magnetic stir bar, place the beaker on a magnetic stir plate, and
stir at slow speed (no visible vortex). Immerse the electrode tips to
just above the rotating stir bar. If using an ISE meter, calibrate the
meter in terms of chloride concentration following the manufacturer's
instructions. If using a pH/mV meter, record the meter reading (mV) as
soon as the reading is stable, but in no case should the time exceed five
minutes after immersing the electrode tips. Prepare a calibration curve
by plotting measured potential (mV) as a function of the logarithm of
chloride concentration. The slope must be 54-60 mV per decade of chloride
concentration. If the slope is not acceptable, the ISE may not be working
properly. For corrective action, consult the ISE operating manual.
7.2 Allow samples and standards to equilibrate to room temperature.
7.3 Prior to and between analyses, rinse the electrodes thoroughly with
reagent water and gently shake off excess water. Low-level measurements are
faster if the electrode tips are first immersed for five minutes in reagent
water.
7.4 Add 50.0 mL of sample and 1.00 mL of ISA to a 100 mL beaker. Add a
Teflon®-coated magnetic stir bar. Place the beaker on a magnetic stir plate and
stir at a slow speed (no visible vortex). Immerse the electrode tip(s) to just
above the rotating stir bar. Record the meter reading (mV or concentration) as
soon as the reading is stable, but in no case should the time exceed five minutes
after immersing the electrode tips. If reading mV, determine chloride
concentration from the calibration curve.
7.5 When analyses have been completed, rinse the electrodes thoroughly
and store them in a 30.0 mg/L chloride standard solution. If the electrodes will
not be used more than one day, drain the internal filling solutions, rinse with
reagent water, and store dry.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Initial Calibration Verification standard (ICV): After performing
the calibration step (7.1), verify calibration by analyzing an ICV. The ICV
contains a known chloride concentration at the mid-range of the calibration
standards and is from an independent source. ICV recovery must be 90-110
percent. If not, the error source must be found and corrected. An acceptable
ICV must be analyzed prior to sample analysis. The ICV also serves as a
laboratory control sample.
8.3 Continuing Calibration Verification standard (CCV): After every 10
samples, and after the final sample, a CCV must be analyzed. The CCV contains
a known chloride concentration at mid-calibration range. CCV recovery must be
90-110 percent. If not, the error source must be found and corrected. If ISE
calibration has changed, all samples analyzed since the last acceptable CCV must
be re-analyzed.
9212 - 4 Revision 0
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8.4 Reagent blank: After the ICV and after every CCV, a reagent blank
must be analyzed. A reagent blank is reagent water mixed 50:1 with ISA. The
indicated reagent blank concentration must be less than 1 mg/L chloride. If not,
the contamination source must be found and corrected. All samples analyzed since
the last acceptable reagent blank must be re-analyzed.
8.5 Matrix spike: Follow the matrix spike protocols presented in Chapter
One. The spike concentration must be 10 times the detection limit and the volume
added must be negligible (less than or equal to one-thousandth the sample aliquot
volume). Spike recovery must be 75-125 percent. If not, samples must be
analyzed by the method of standard additions.
9.0 METHOD PERFORMANCE
9.1 In a single-laboratory evaluation, a series of standards with known
chloride concentrations was analyzed with a chloride ISE. Measurements were
conducted over three consecutive days using an Orion 9609 chloride combination
ISE connected to an Orion 940 ISE meter. A two-point calibration (4.CO and 40.0
mg/L chloride) was performed prior to analysis. The results are listed in Table
2.
9.2 In the same study, six groundwater samples were spiked with chloride
at four different concentrations and were measured with the chloride ISE. (The
groundwater samples initially contained 1.3-23 mg/L chloride.) Each spiked
sample was analyzed at each concentration and the mean recoveries and RSDs are
given in Table 3.
9.3 A 50 g portion of soil, which initially contained 17.3 mg/kg
chloride, was spiked with 25.0 mg/kg chloride to obtain an anion concentration
in a single extract volume within the linear range of the ISE. The extract was
then analyzed for chloride using this ISE method, and 109% of the soil spike was
recovered.
10.0 REFERENCES
1. Franson, Mary Ann H., Ed. Standard Methods for the Examination of
Water and Wastewater, 18th Edition. American Public Health Association,
Washington, DC, 1992.
2. Model 96-17 Chloride Combination Electrode Instruction Manual. Orion
Research, Inc., Boston, MA, 1988.
3. Miller, E.L., Waltman, D.W., and Hillman, D.C. Single-Laboratory
Evaluation of Fluoride, Chloride, Bromide, Cyanide, and Nitrate Ion-Selective
Electrodes for Use in SW-846 Methods. Lockheed Engineering and Sciences Company
for Environmental Monitoring Systems Laboratory, U.S. EPA. September 1990.
EPA/600/X-90/221.
4. Cotton, F. Albert, and Geoffrey Wilkinson; Advanced Inorganic
Chemistry, 2nd Edition; Interscience Publishers, New York, NY; 1966.
5. Weast Robert C., Ed.; CRC Handbook of Chemistry and Physics, 58th
Edition; CRC Press, Inc., Cleveland, Ohio; 1977.
9212 - 5 Revision 0
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6. Kolthoff, I.M., E.B. Sandell, E.J. Meehan, and Stanley Bruckenstein;
Quantitative Chemical Analysis, 4th Edition; The MacMillan Company, New York, NY;
1969.
i
9212 - 6 Revision 0
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Table 1. Chloride ISE Interferences
Analyte
Concentration
(mg/L)
20.0
200.0
20.0
200.0
20.0
200.0
200.0
200.0
20.0
Interference
None
None
300 mg/L Fe+3
300 mg/L Fe+3
300 mg/L Al+3
300 mg/L Al+3
0.01 M H2S04
200 mg/L CN' +
0.01 M H2S04
200 mg/L S'2 +
0.01 M H2S04
Measured
Concentration
(mg/L)
19.9
200.0
19.6
183.0
19.3
175.0
201.0
198.0
19.9
RSD
(%)
2.2
1.3
3.0
4.5
3.3
6.2
1.5
1.0
0.5
Table 2. Results from a single-laboratory accuracy evaluation
of a chloride ISE
Chloride
Concentration
(mg/L)
0.400
1.00
2.00
4.00
10.0
20.0
40.0
100
200
400
1,000
Chloride
Detected
(mg/L)
0.64
1.32
2.07
4.0
10.0
19.4
40.0
100
201
390
970
Chloride
Recovery
(percent)
160
132
104
100
100
97
100
100
101
99
97
Rel. Std.
Deviation
(percent)
21
9
4
3
4
7
3
4
1
4
3
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Table 3. Mean Spike Recoveries of Chloride in 6 Groundwater Samples
Analyte Spike Spike Rel. Std.
Concentration Recovery Deviation
(mg/L) (percent) (percent)
2.00 107 8.2
3.00 95 3.6
5.00 93 3.6
10.0 102 4.3
i
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METHOD 9212
POTENTIQMETRIC DETERMINATION OF CHLORIDE
IN AQUEOUS SAMPLES WITH ION-SELECTIVE ELECTRODE
>
r
7.1.1 - 7.1.2
Calibrate Chloride
ISE.
^
r
7.2 Allow standards
to equilibrate to
room temperature.
^
r
7.3 Rinse
eletrodes.
i
r
7.4 Measure
concentration using
electrode meter
and calculate
concentration.
i
r
7.5 Drain
reference electrode
and clean.
^
r
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METHOD 9213
POTENTIOHETRIC DETERMINATION OF CYANIDE
IN AQUEOUS SAMPLES AND DISTILLATES WITH ION-SELECTIVE ELECTRODE
1.0 SCOPE AND APPLICATION
1.1 This method can be used for measuring free (non-complexed) cyanide
and hydrocyanic acid in drinking waters, natural surface waters, domestic and
industrial wastewaters, and in soil extracts (ASTM methods D4646-87, D5233-92 or
D3987-85). The method may also be used as the determinative step for total
cyanide in distillate following the distillation in Method 9010.
1.2 The method detection limit is 0.05 mg/L. Cyanide concentrations from
0.01 to 10 mg/L may be measured. However, using a linear calibration, results
less than 0.05 mg/L may be biased up to approximately 120 percent high.
1.3 ISEs must be used carefully, and results must be interpreted
cautiously, since an ISE may be affected by numerous analytical interferences
which may either increase or decrease the apparent analyte concentration, or
which may damage the ISE. Effects of most interferences can be minimized or
eliminated by adding appropriate chemical reagents to the sample. Obtaining the
most accurate results, therefore, requires some knowledge of the sample
composition.
NOTE: ISE manufacturers usually include a list of interferences in
the instruction manual accompanying an ISE, along with recommended
methods for minimizing or eliminating effects of these
interferences.
1.4 The cyanide ISE should not be used in solutions containing more than
30 mg/L of free cyanide, because such solutions will dissolve the ISE sensor
silver compounds.
2.0 SUMMARY OF METHOD
2.1 Cyanide is determined potentiometrically using a cyanide ion-
selective electrode (ISE) in conjunction with a double-junction reference
electrode and a pH meter with an expanded millivolt scale or an ISE meter capable
of being calibrated directly in terms of cyanide concentration.
2.2 Standards and samples are mixed 100:1 with an ionic strength
adjustment solution (ISA). Calibration is performed by analyzing a series of
standards and plotting mV vs. cyanide concentration on semilog paper or by
calibrating the ion meter directly in terms of cyanide concentration.
3.0 INTERFERENCES
3.1 Transition metal cations interfere by forming very stable complexes
with cyanide which are not measured by the cyanide ISE. For example, copper(II)
ions interfere with the cyanide determination by oxidizing cyanide to cyanogen
gas and precipitating copper(I) cyanide. Because these complexes are very stable
and their rate of dissociation is slow (days) at room temperature (25°C), this
9213 - 1 Revision 0
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method is restricted to the analysis of free cyanide and hydrocyanic acid in
undistilled samples. Total cyanide may be determined in samples distilled as per
Method 9010. (Distillation removes interferants and breaks down metal-analyte
complexes and insoluble salts.)
3.2 Several anions and one acid were tested for cyanide interference.
(See Table 1.) As shown, hydrogen ions (acid) interfere by forming hydrocyanic
acid. Hydrogen ions are removed by adding ISA.
3.3 Sulfide interferes with the determination by reacting directly with
the ISE. This interference can be removed after adding ISA by adding and well
mixing small amounts (about 0.05 g) of powdered lead carbonate until the added
powder remains white.
3.4 Temperature changes affect electrode potentials. Using an ISE
calibrated at 22 C, a 1.00 mg/L cyanide solution was measured as 0.98 mg/L at 22
°C and 0.64 mg/L at 32°C (see Ref. 4). Therefore, standards and samples must be
equilibrated at the same temperature ( + 1°C).
3.5 The user should be aware of the potential of intereferences from
colloidal substances and that, if necessary, the samples may be filtered.
4.0 APPARATUS AND MATERIALS
4.1 pH/mV meter capable of reading to 0.1 mV or an ISE meter.
4.2 Cyanide ISE (Orion 9406 or equivalent) and double-junction reference
electrode (Orion 9002 or equivalent).
4.3 Thermally isolated magnetic stirrer, Teflon®-coated stir bar, and
stopwatch.
4.4 Volumetric flask, 100 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 reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 ISA solution (10M NaOH): Dissolve 40 g of sodium hydroxide in
sufficient reagent water to make 100 mL of solution. Cool, and store in a
polyethylene bottle.
CAUTION: This solution is extremely corrosive.
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5.4 Powdered lead carbonate [(PbC03)2 • Pb(OH)2]: use in approximately
0.05 g increments to remove sulfide interference. (See Step 3.3.)
5.5 Cyanide calibration stock solution (1,000 mg/L CN"): Dissolve
0.2504 g of potassium cyanide (dried two hours at 110°C and stored in a
desiccator) in reagent water, add 1.0 ml of ISA and dilute to 100.0 ml in a
volumetric flask. Store in a clean bottle.
5.6 Cyanide standard solution (100.0 mg/L CN"): Dilute 10.00 ml of
1,000 mg/L cyanide calibration stock solution to 100.0 mL with reagent water in
a volumetric flask.
5.7 Cyanide calibration standards: Prepare a series of calibration
standards by diluting the 100.0 mg/L cyanide standard. A suitable series is
given in the table below.
mL of 100.0 mg/L Concentration when Diluted
CM' Solution to 50.0 mL (mg/L CN')
0.0150 0.0300
0.0500 0.100
0.150 0.300
0.500 1.00
1.50 3.00
5.8 Sodium hydroxide solution (50%), NaOH. Commercially available.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 Samples must be preserved by adding 50% sodium hydroxide until the
pH is greater than or equal to 12 at the time of collection and should be chilled
to 4°C. When properly preserved, cyanide samples can be stored for up to 14 days
prior to analysis.
7.0 PROCEDURE
7.1 Calibration
7.1.1 When using a cyanide ISE and a separate double-junction
reference electrode, ensure that reference electrode inner and outer
chambers are filled with solutions recommended by the manufacturer.
Equilibrate the electrodes for at least one hour in a 0.300 mg/L cyanide
standard before use.
7.1.2 Calibrate the cyanide ISE using standards that narrowly
bracket the expected sample concentration. If the sample concentration is
unknown, calibrate with 0.100 mg/L and 1.00 mg/L cyanide standards. Add
50.0 mL of standard and 0.50 mL of ISA to a 100 mL beaker. Add a Teflon®-
coated magnetic stir bar, place the beaker on a magnetic stir plate, and
9213 - 3 Revision 0
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stir at slow speed (no visible vortex). Immerse the electrode tips to
just above the rotating stir bar. If using an ISE meter, calibrate the
meter in terms of cyanide concentration following the manufacturer's
instructions. If using a pH/mV meter, record the meter reading (mV) as
soon as the reading is stable, but in no case should the time exceed five
minutes after immersing the electrode tips. Prepare a calibration curve
by plotting measured potential (mV) as a function of the logarithm of
cyanide concentration. The slope must be 54-60 mV per decade of cyanide
concentration. If the slope is not acceptable, the ISE may not be working
properly. For corrective action, consult the ISE operating manual.
7.2 Allow samples and standards to equilibrate to room temperature.
7.3 Prior to and between analyses, rinse the electrodes thoroughly with
reagent water and gently shake off excess water. Low-level measurements are
faster if the electrode tips are first immersed five minutes in reagent water.
7.4 Add 50.0 ml of sample and 0.50 ml of ISA to a 100 ml beaker. Add a
Teflon®-coated magnetic stir bar. Place the beaker on a magnetic stir plate and
stir at a slow speed (no visible vortex). Immerse the electrode tips to just
above the rotating stir bar. Record the meter reading (mV or concentration) as
soon as the reading is stable, but in no case should the time exceed five minutes
after immersing the electrode tips. If reading mV, determine cyanide
concentration from the calibration curve.
7.5 When analyses have been completed, rinse the electrodes thoroughly
and store them in a 0.30 mg/L cyanide standard solution. If the electrodes will
not be used more than one day, drain the reference electrode internal filling
solutions, rinse with reagent water, and store dry.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Initial Calibration Verification standard (ICV): After performing
the calibration step (7.1), verify calibration by analyzing an ICV. The ICV
contains a known cyanide concentration at the mid-range of the calibration
standards and is from an independent source. ICV recovery must be 90-110
percent. If not, the error source must be found and corrected. An acceptable
ICV must be analyzed prior to sample analysis. The ICV also serves as a
laboratory control sample.
8.3 Continuing Calibration Verification standard (CCV): After every 10
samples, and after the final sample, a CCV must be analyzed. The CCV contains
a known cyanide concentration at mid-calibration range. CCV recovery must be 90-
110 percent. If not, the error source must be found and corrected. If the ISE
calibration has changed, all samples analyzed since the last acceptable CCV must
be re-analyzed.
8.4 Reagent blank: After the ICV and after every CCV, a reagent blank
must be analyzed. A reagent blank is reagent water mixed 100:1 with ISA. The
indicated reagent blank concentration must be less than 0.03 mg/L cyanide. If
not, the contamination source must be found and corrected. All samples analyzed
since the last acceptable reagent blank must be re-analyzed.
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8.5 Matrix spike: Follow the matrix spike protocols presented in Chapter
One. The spike concentration must be 10 times the detection limit and the volume
added must be negligible (less than or equal to one-thousandth the sample aliquot
volume). Spike recovery must be 75-125 percent. If not, samples must be
analyzed by the method of standard additions.
9.0 METHOD PERFORMANCE
9.1 In a single-laboratory evaluation, a series of standards with known
cyanide concentrations was analyzed with a cyanide ISE. Measurements were
obtained over three consecutive days using an Orion 9406 cyanide ISE and an Orion
9002 double-junction reference electrode connected to an Orion 940 ISE meter.
A two-point calibration (0.20 and 2.0 mg/L cyanide) was performed prior to
analysis. The results are listed in Table 2.
9.2 In the same study, three groundwater samples were spiked with cyanide
at four different concentrations and were measured with the cyanide ISE. (The
groundwater samples initially contained <0.1 mg/L cyanide.) Each spiked sample
was analyzed at each concentration and the mean recoveries and RSDs are given in
Table 3.
9.3 A 50g portion of soil, which initially contained <0.1 mg/kg cyanide,
was spiked with 2.50 mg/kg cyanide to obtain an anion concentration in a single
extract volume within the linear range of the ISE. The extract was then analyzed
for cyanide using this ISE method, and 92% of the soil spike was recovered.
10.0 REFERENCES
1. Franson, Mary Ann H., Ed. Standard Methods for the Examination of
Water and Wastewater, 18th Edition. American Public Health Association,
Washington, DC, 1992.
2. Model 94-06 Cyanide Electrode Instruction Manual. Orion Research,
Inc., Boston, MA, 1986.
3. Miller, E.L., Waltman, D.W., and Hillman, D.C. Single-Laboratory
Evaluation of Fluoride, Chloride, Bromide, Cyanide, and Nitrate Ion-Selective
Electrodes for Use in SW-846 Methods. Lockheed Engineering and Sciences Company
for Environmental Monitoring Systems Laboratory, U.S. EPA. September 1990.
EPA/600/X-90/221.
4. Cotton, F. Albert, and Geoffrey Wilkinson; Advanced Inorganic
Chemistry, 2nd Edition; Interscience Publishers, New York, NY; 1966.
5. Kolthoff, I.M., E.B. Sandell, E.J. Meehan, and Stanley Bruckenstein;
Quantitative Chemical Analysis, 4th Edition; The MacMillan Company, New York, NY;
1969.
9213 - 5 Revision 0
January 1995
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Table 1. Cyanide ISE Interferences
Analyte
Concentration
(mg/L)
0.200
2.00
2.00
0.200
0.200
2.00
Interference
None
None
0.01 M H2S04
200 mg/L CT
200 mg/L Br"
20 mg/L S'2 + 0.05
g PbCo3
Measured
Concentration
(mg/L)
0.201
2.00
<0.05
0.204
0.200
2.02
RSD
(%)
1.0
0.7
*
2.0
2.7
1.6
* Single Measurement
Table 2. Results From a Single-Laboratory Accuracy
Evaluation of a Cyanide ISE
Cyanide
Concentration
(mg/L)
0.0100
0.0200
0.0500
0.100
0.200
0.500
1.00
2.00
5.00
10.0
Cyanide
Detected
(mg/L)
0.0217
0.0340
0.0520
0.103
0.198
0.48
1.03
2.02
5.00
9.9
Cyanide
Recovery
(percent)
217
170
104
103
99
96
103
101
100
99
Rel. Std.
Deviation
(percent)
27
13
8
6
3
6
6
3
6
4
9213 - 6
Revision 0
January 1995
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Table 3. Mean Spike Recoveries of Cyanide in 3 Groundwater Samples
Analyte Spike Spike Rel. Std.
Concentration Recovery Deviation
(mg/L) (percent) (percent)
0.0500 115 22.3
0.150 103 7.6
0.300 98 8.3
1.00 103 1.1
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January 1995
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METHOD 9213
POTENTIOMETRIC DETERMINATION OF CYANIDE
IN AQUEOUS SAMPLES WITH ION-SELECTIVE ELECTRODE
^
f
7.1.1 - 7.1.2
Calibrate Cyanide
ISE.
•^
r
7.2 Allow standards
to equilibrate to
room temperature.
^
r
7.3 Rinse
eletrodes.
^
r
7.4 Measure
concentration using
electrode meter
and calculate
concentration.
i
r
7.5 Drain
reference electrode
and clean.
\
r
9213 - 8
Revision 0
January 1995
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METHOD 9214
POTENTIOMETRIC DETERMINATION OF FLUORIDE
IN AQUEOUS SAMPLES WITH ION-SELECTIVE ELECTRODE
1.0 SCOPE AND APPLICATION
1.1 This method can be used for measuring total solubilized fluoride in
drinking waters, natural surface waters, groundwaters, domestic and industrial
wastewaters, and in soil extracts (ASTM methods D4646-87, D5233-92 or D3987-85).
NOTE: This method is for the analysis of simple fluoride ion rather
than total fluoride, as analysis using the ion-selective electrode
is not preceded by a distillation step.
1.2 The method detection limit is 0.5 mg/L. Fluoride concentrations from
0.025 to 500 mg/L may be measured. However, using a linear calibration, results
less than 0.5 mg/L may be biased up to approximately 160% high.
1.3 ISEs must be used carefully and results must be interpreted
cautiously, since an ISE may be affected by numerous analytical interferences
which may either increase or decrease the apparent analyte concentration, or
which may damage the ISE. Effects of most interferences can be minimized or
eliminated by adding appropriate chemical reagents to the sample. Obtaining the
most accurate results, therefore, requires some knowledge of the sample
composition.
NOTE: ISE manufacturers usually include a list of interferences in
the instruction manual accompanying an ISE, along with recommended
methods for minimizing or eliminating effects of these
interferences.
2.0 SUMMARY OF METHOD
2.1 Total solubilized fluoride is determined potentiometrically using a
fluoride ion-selective electrode (ISE) in conjunction with a standard single-
junction reference electrode, or a fluoride combination ISE, and a pH meter with
an expanded millivolt scale or an ISE meter capable of being calibrated directly
in terms of fluoride concentration.
2.2 Standards and samples are mixed 1:1 with a total ionic strength
adjustment buffer (TISAB). TISAB adjusts ionic strength, buffers pH to 5-5.5,
and contains a chelating agent to break up metal-fluoride complexes. Calibration
is performed by analyzing a series of standards and plotting mV vs. fluoride
concentration on semilog paper or by calibrating the ion meter directly in terms
of fluoride concentration.
3.0 INTERFERENCES
3.1 Polyvalent cations (e.g., Fe+3 and Al"1"3) interfere by forming
complexes with fluoride which are not measured by the fluoride ISE. (See Table
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1.) The results in Table 1 are in agreement with the fact that aluminum-fluoride
complex ions are approximately ten times more stable than corresponding
iron(III)-fluoride complex ions. As the aluminum concentration increases, more
fluoride is consumed to form the metal-fluoride complex. Adding TISAB, which
contains a strong chelating agent, eliminates this interference by complexing
polyvalent cations.
3.2 Sample pH is critical. Hydroxide is a significant interferant at
concentrations ten times the fluoride concentration. This interference is
avoided by adding TISAB which buffers the sample at a pH of 5-5.5. At low pH
values, fluoride forms bifluoride (HF2~) which is not detected by the fluoride
ISE. Again, adding TISAB prevents this interference by buffering the pH.
3.3 Temperature changes affect electrode potentials. Using an ISE
calibrated at 22°C, a 20.0 mg/L fluoride solution was measured as 20.3 mg/L at
22°C and 13.6 mg/L at 32°C (see Ref. 4). Therefore, standards and samples must
be equilibrated at the same temperature (+ 1°C).
3.4 The user should be aware of the potential of intereferences from
colloidal substances and that, if necessary, the samples may be filtered.
4.0 APPARATUS AND MATERIALS
4.1 pH/mV meter capable of reading to 0.1 mV or an ISE meter.
4.2 Combination fluoride ISE (Orion 9609 or equivalent), or separate
fluoride ISE (Orion 9409 or equivalent) and reference electrode (Orion 9001 or
equivalent) prepared for use as described in owner's manual.
4.3 Thermally isolated magnetic stirrer, Teflon®-coated stir bar, and
stopwatch.
4.4 Volumetric flask, 1,000 mL.
4.5 Polyethylene labware.
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 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Sodium hydroxide solution (5M NaOH): Dissolve 200 g of NaOH in
sufficient reagent water to make 1 L of solution. Store in a tightly sealed
polyethylene bottle.
9214 - 2 Revision 0
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CAUTION: This solution is extremely corrosive.
5.4 Glacial acetic acid (CH3C02H).
5.5 Sodium chloride (NaCl).
5.6 CDTAor 1,2-cyclohexanedinitrilo-tetraacetic acid (C6H10[N(CH2C02H)2]2
• H20).
5.7 TISAB solution
5.7.1 To approximately 500 mL of reagent water add 57.0 ml of
glacial acetic acid, 58.0 g of sodium chloride, and 4.00 g of CDTA. Stir
to dissolve and cool to room temperature. Adjust the solution pH to
between 5.0 and 5.5 with 5M NaOH (about 150 ml will be required).
Transfer the solution to a 1,000 ml volumetric flask and dilute to the
mark with reagent water. Transfer the solution to a clean polyethylene
bottle.
5.7.2 Alternatively, TISAB solution is available commercially (Orion
940999 or equivalent).
5.8 Fluoride calibration stock solution (1,000 mg/L F"): Dissolve
0.2210 g of sodium fluoride (NaF, dried two hours at 110°C and stored in a
desiccator) in reagent water and dilute to 100 mL in a polyethylene volumetric
flask. Store in a clean polyethylene bottle.
5.9 Fluoride standard solution (100 mg/L F"): Dilute 10.0 mL of 1,000
mg/L fluoride calibration stock solution to 100 mL with reagent water in a
polyethylene volumetric flask.
5.10 Fluoride calibration standards: Prepare a series of calibration
standards by diluting the 100 mg/L fluoride standard. A suitable series is given
in the table below.
mL of 100 mg/L Concentration when Diluted
F" Solution to 50.0 mL (mg/L F')
0.0500 0.100
0.150 0.300
0.500 1.00
1.50 3.00
5.00 10.0
15.0 30.0
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
9214 - 3 Revision 0
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6.2 Samples must be stored and handled in polyethylene containers.
Samples should be stored at 4°C.
7.0 PROCEDURE
7.1 Calibration
7.1.1 If using a fluoride combination ISE, ensure that the ISE is
filled with the solution recommended by the manufacturer. Change the
solution if the ISE has not been used for a week. If using a fluoride ISE
and a separate reference electrode, ensure that the reference electrode is
filled with the solution recommended by the manufacturer. In either case,
equilibrate the electrodes for 24 hours in a 10.0 mg/L fluoride standard
before use.
7.1.2 Calibrate the fluoride ISE using standards that narrowly
bracket the expected sample concentration. If the sample concentration is
unknown, calibrate with 1.00 mg/L and 10.0 mg/L fluoride standards. Add
20.0 mL of standard and 20.0 mL of TISAB to a 50 mL polyethylene beaker.
Add a Teflon®-coated magnetic stir bar, place the beaker on a magnetic
stir plate, and stir at slow speed (no visible vortex). Immerse the
electrode tips to just above the rotating stir bar. If using an ISE
meter, calibrate the meter in terms of fluoride concentration following
the manufacturer's instructions. If using a pH/mV meter, record the meter
reading (mV) as soon as the reading is stable, but in no case should the
time exceed five minutes after immersing the electrode tips. Prepare a
calibration curve by plotting measured potential (mV) as a function of the
logarithm of fluoride concentration. The slope must be 54-60 mV per
decade of fluoride concentration. If the slope is not acceptable, the ISE
may not be working properly. For corrective action, consult the ISE
operating manual.
NOTE: Use only plasticware (polyethylene) when preparing and
analyzing fluoride samples.
7.2 Allow samples and standards to equilibrate to room temperature.
7.3 Prior to and between analyses, rinse the electrode thoroughly with
reagent water and gently shake off excess water. Low-level measurements are
faster if the electrode tips are first immersed for five minutes in reagent
water.
7.4 Add 20.0 mL of sample and 20.0 mL of TISAB to a 50 mL polyethylene
beaker. Add a Teflon®-coated magnetic stir bar. Place the beaker on a magnetic
stir plate and stir at a slow speed (no visible vortex). Immerse the electrode
tips to just above the rotating stir bar. Record the meter reading (mV or
concentration) as soon as the reading is stable, but in no case should the time
exceed five minutes after immersing the electrode tips. If reading mV, determine
fluoride concentration from the calibration curve.
7.5 When analyses have been completed, rinse the electrodes thoroughly
and store them in a 10.0 mg/L fluoride standard solution. If the electrodes will
9214 - 4 Revision 0
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not be used more than one day, drain the internal filling solution, rinse with
reagent water, and store dry.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Initial Calibration Verification standard (ICV): After performing
the calibration step (7.1), verify calibration by analyzing an ICV. The ICV
contains a known fluoride concentration at mid-range of the calibration standards
and is from an independent source. ICV recovery must be 90-110 percent. If not,
the error source must be found and corrected. An acceptable ICV must be analyzed
prior to sample analysis. The ICV also serves as a laboratory control sample.
8.3 Continuing Calibration Verification standard (CCV): After every 10
samples, and after the final sample, a CCV must be analyzed. The CCV contains
a known fluoride concentration at mid-calibration range. CCV recovery must be
90-110 percent. If not, the error source must be found and corrected. If ISE
calibration has changed, all samples analyzed since the last acceptable CCV must
be re-analyzed.
8.4 Reagent blank: After the ICV and after every CCV, a reagent blank
must be analyzed. A reagent blank is reagent water mixed 1:1 with TISAB. The
indicated reagent blank concentration must be less than 0.1 mg/L fluoride. If
not, the contamination source must be found and corrected. All samples analyzed
since the last acceptable reagent blank must be re-analyzed.
8.5 Matrix spike: Follow the matrix spike protocols presented in Chapter
One. The spike concentration must be 10 times the detection limit and the volume
added must be negligible (less than or equal to one-thousandth the sample aliquot
volume). Spike recovery must be 75-125 percent. If not, samples must be
analyzed by the method of standard additions.
9.0 METHOD PERFORMANCE
9.1 In a single-laboratory evaluation, a series of standards with known
fluoride concentrations was analyzed with a fluoride ISE. Measurements were
conducted over three consecutive days using an Orion 9609 fluoride combination
ISE connected to an Orion 940 ISE meter. A two-point calibration (1.00 and 10.0
mg/L fluoride) was performed prior to analysis. The results are listed in Table
2.
9.2 In the same study, 12 groundwater samples and six extraction
procedure (EP, Method 1310) soil leachate samples were spiked with fluoride at
four different concentrations and were measured with the fluoride ISE. (The
groundwater samples initially contained 0.1-2 mg/L fluoride and the EP leachates
initially contained 0.1-8 mg/L fluoride.) Each spiked sample was analyzed at
each concentration and the mean recoveries and RSDs are listed in Table 3.
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10.0 REFERENCES
1. Franson, Mary Ann H., Ed. Standard Methods for the Examination of
Water and Wastewater, 18th Edition. American Public Health Association,
Washington, DC, 1992.
2. Model 96-09 Fluoride Combination Electrode Instruction Manual. Orion
Research, Inc., Boston, MA, 1988.
3. Methods for Chemical Analysis of Water and Wastes. U.S.
Environmental Protection Agency, Cincinnati, Ohio, 1983.
4. Miller, E.L., Waltman, D.W., and Hillman, D.C. Single-Laboratory
Evaluation of Fluoride, Chloride, Bromide, Cyanide, and Nitrate Ion-Selective
Electrodes for Use in SW-846 Methods. Lockheed Engineering and Sciences Company
for Environmental Monitoring Systems Laboratory, U.S. EPA. September 1990.
EPA/600/X-90/221.
5. Cotton, F. Albert, and Geoffrey Wilkinson; Advanced Inorganic
Chemistry, 2nd Edition; Interscience Publishers, New York, NY; 1966.
6. Weast, Robert C., Ed.; CRC Handbook of Chemistry and Physics, 58th
Edition; CRC Press, Inc., Cleveland, Ohio; 1977.
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Table 1. Fluoride ISE Interferences
Analyte
Concentration
(mg/L)
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
Interference
None
1 mg/L Al+3
5 mg/L Al+3
25 mg/L Al+3
50 mg/L Al+3
1 mg/L Fe+3
5 mg/L Fe+3
50 mg/L Fe+3
Measured
Concentration
(mg/L)
0.95
0.97
0.86
0.51
0.41
1.00
1.02
0.92
RSD
t °/\
(/o)
2.0
3.1
3.9
*
4.9
3.9
3.8
3.2
* Single measurement
Table 2. Results from a Single-Laboratory Accuracy
Evaluation of a Fluoride ISE
Fluoride
Concentration
(mg/L)
0.0250
0.0500
0.125
0.250
0.500
2.50
5.00
25.0
50.0
250.
500.
Fluoride
Detected
(mg/L)
0.066
0.085
0.164
0.31
0.51
2.43
5.0
25.2
53.
260.
530.
Fluoride
Recovery
(%)
264
170
131
125
103
97
101
101
105
105
107
Rel. Std.
Deviation
IV \
(/o)
19
11
10
7
2
2
2
2
3
3
3
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Table 3. Mean Spike Recoveries for 12 Groundwater Samples and
6 EP Leachate Samples
Spike Groundwater EP Leachate
Fluoride Fluoride Rel. Std. Fluoride Rel. Std.
Added Recovered Deviation Recovered Deviation
(mg/L) (percent) (percent) (percent) (percent)
0.5 92 5.3 100 20
1.5 92 3.3 92 9.7
3.5 93 3.0 91 5.5
8.5 96 2.6 91 4.1
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METHOD 9214
POTENTIOMETRIC DETERMINATION OF FLUORIDE
IN AQUEOUS SAMPLES WITH ION-SELECTIVE ELECTRODE
Start
J)
^
r
7.1.1 - 7.1.2
Calibrate Fluoride
ISE.
•^
r
7.2 Allow standards
to equilibrate to
room temperature.
>
r
7.3 Rinse
eletrodes.
>
r
7.4 Measure
concentration using
electrode meter
and calculate
concentration.
•^
i
7.5 Drain
reference electrode
and clean.
>
r
9214 - 9
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METHOD 9215
POTENTIOMETRIC DETERMINATION OF SULFIDE
IN AQUEOUS SAMPLES AND DISTILLATES WITH ION-SELECTIVE ELECTRODE
1.0 SCOPE AND APPLICATION
1.1 This method can be used for measuring total sulfide in a distilled
sample. The method is meant to be used as an alternate determinative step
following the distillation in either SW-846 Methods 9030 or 9031.
1.2 This method must not be used for undistilled samples because of
possible mercury and silver ion interferences. Also, the ISE only responds to
free sulfide dianion and will not detect sulfide in complexes. Therefore, this
method would provide low recoveries for undistilled samples.
1.3 The method detection limit is 1.0 mg/L. Sulfide concentrations from
0.1 to 12,000 mg/L may be measured. However, when a linear calibration is used,
results less than 1 mg/L may be biased up to approximately 90 percent low.
2.0 SUMMARY OF METHOD
2.1 The distillations in Methods 9030 and/or 9031 are performed, except
that the scrubber solution is sulfide anti-oxidant buffer (SAOB), with ascorbic
and salicylic acids added as oxygen scavengers. The distillates are then
analyzed potentiometrically using a sulfide ion-selective electrode (ISE) in
conjunction with a double-junction reference electrode and a pH meter with an
expanded millivolt scale or an ISE meter capable of being calibrated directly in
terms of sulfide concentration.
2.2 This method is an alternative to the iodometric titration procedure,
where a solution of thiosulfate is standardized against the thiosulfate solution,
and the sulfide standard is standardized against the iodine solution/thiosulfate
solution (which requires daily calibration itself). This method allows for
standardizing the sulfide calibration standards by a potentiometric titration
with standardized silver nitrate using the sulfide ISE as the working electrode.
Silver nitrate solutions are stable when stored properly and are easily
standardized against sodium chloride.
2.3 The key to acceptable recoveries is the use of the proper apparatus
and careful assembly of the distillation apparatus prior to distillation. If
these steps are not taken, low recoveries will result.
3.0 INTERFERENCES
3.1 Since this method may only be used for distillates, which will have
a pH greater than 12, there are no expected interferences. As shown in Table 1,
the data indicate that neither silver, mercury, nor humic acid interfere with the
electrode. (An interference would be indicated by a positive response.)
3.2 The effect of sulfur(+4) compounds (which interfere with the
titrimetric method) in the absorbing solution on the sulfide ISE was tested
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(e.g., sulfite or thiosulfate). The electrode did not respond to either
compound. A 110 mg/L sulfide solution was spiked with 100 mg/L sulfide and 100
mg/L thiosulfate. The measured concentrations for the unspiked and spiked
samples were identical (114 mg/L).
3.3 Temperature changes affect electrode potentials. Standards and
samples must be equilibrated at the same temperature (+ 1°C).
3.4 The user should be aware of the potential of intereferences from
colloidal substances and that, if necessary, the samples may be filtered.
4.0 APPARATUS AND MATERIALS
4.1 pH/mV meter capable of reading to 0.1 mV or an ISE meter.
4.2 Sulfide ISE (Orion 9416 or equivalent) and double-junction reference
electrode (Orion 902000 or equivalent).
4.3 Thermally isolated magnetic stirrer, Teflon®-coated stir bar, and
stopwatch.
4.4 Volumetric flask, 125 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 reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Sulfide anti-oxidant buffer (SAOB). Dissolve 80 g NaOH, 320 g sodium
salicylate and 72 g ascorbic acid in 1 L reagent water. Prepare fresh weekly.
5.4 Standard sodium chloride solution (0.100N). Dissolve 5.84 g NaCl
(dried for 2 hours at 140"C) in water and dilute to 1.00 L with reagent water.
5.5 Sodium hydroxide (6N), NaOH. Dissolve 240 g of NaOH in 1 L of
reagent water. Keep tightly closed.
5.6 Potassium chromate indicator solution. Dissolve 50 g K2Cr04 in a
little reagent water. Add AgN03 solution until a definite red precipitate is
formed. Let stand 12 hrs, filter, and dilute to 1 L.
5.7 Standard silver nitrate solution (0.10N). Dissolve 16.99 g of AgN03
(dried for 2 hours at 150"C) in reagent water and dilute to 1.00 L. Store in a
brown bottle. Standardize weekly against standard sodium chloride solution.
5.8 Sodium sulfide nonahydrate, Na2S • 9H20. For the preparation of
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sulfide standard solutions to be used for calibration curves. Standards .must be
prepared between pH 9 and pH 11. Protect standards from exposure to oxygen by
preparing them without headspace. These standards are unstable and should be
prepared daily.
5.9 Sodium sulfide (2% and 10%), Na2S.
5.10 Ammonia, NH3: concentrated.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 All original, undistilled samples should be handled as described in
Section 6 of Methods 9030 and/or 9031. Samples should be stored at 4°C prior to
analysis.
6.3 The distilled samples should be analyzed with the ISE immediately
following distillation, distillates that are not analyzed immediately should be
stored in a sealed flask at 4°C.
7.0 PROCEDURE
7.1 Replace the scrubber solution in Method 9030 or 9031 with 10.0 mL
SAOB solution and 40.0 mL reagent water. Proceed to distill samples as described
in those methods.
7.2 Standardization of silver nitrate
7.2.1 Add 10.00 mL of 0.100N NaCl and 40 mL reagent water to a 125
mL flask. Adjust pH to 7-10 with dilute NaOH solution. Add 1.0 mL
potassium chromate indicator. Titrate with silver nitrate solution to a
pinkish-yellow endpoint. Be consistent with endpoint recognition. Repeat
with a reagent blank (water and indicator). Calculate the normality of
the silver nitrate as follows:
N AgN03 = (A - B) x N(NaC1)
10.00 mL
A = mL titration for NaCl
B = mL titration for reagent blank
7.3 Standardization of sulfide standards
7.3.1 From the sodium sulfide salt, prepare standards with nominal
concentrations of 10, 100, and 1,000 ppm sulfide in a matrix of 20% SAOB.
Standardize each solution immediately prior to calibrating the ISE. The
standards may be calibrated by iodometric titration or by potentiometric
titration as described below.
7.3.2 The titration is monitored with the combination silver/sulfide
electrode (silver-coated platinum ring sensing electrode with a
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syilver/silver chloride reference electrode). Prior to use the electrode
is conditioned by soaking in 2% sodium sulfide for 5 minutes, soaking in
10% sodium sulfide until the brownish layer becomes black, rinsing with
reagent water, and cleaning with a soft cloth. After conditioning, the
electrode is connected to the pH/mV meter. 20 ml of a sulfide standard
(or suitable quantity to get accurate titration) and 1 ml concentrated
ammonia are pipetted into a titration vessel. The electrode is inserted
and the potential recorded. The sample is titrated with the standardized
silver nitrate until a potential of 100 mV is attained. The potential is
recorded after each titrant addition. The equivalence point is determined
from the first derivative of the titration curve. The sulfide
concentration is then calculated as follows:
sulfide (ppm) = A x B x 16,000 mg/eq
C
A = ml silver nitrate
B = normality of silver nitrate
C = ml of sulfide standard
7.4 Calibration of Sulfide ISE and Meter
7.4.1 Calibrate the sulfide ISE using the 10, 100, and 1,000 mg/L
sulfide standards. The standards must be freshly standardized. Add 25.0
ml of standard into a 50 ml beaker. Add a Teflon®-coated magnetic stir
bar, place the beaker on a magnetic stir plate, and stir at slow speed (no
visible vortex). Immerse the electrode tips to just above the rotating
stir bar. If using an ISE meter, calibrate the meter in terms of sulfide
concentration following the manufacturer's instructions. If using a pH/mV
meter, record the meter reading (mV) as soon as the reading is stable, but
in no case should the time exceed five minutes after immersing the
electrode tips. Prepare a calibration curve by plotting measured
potential (mV) as a function of the logarithm of sulfide concentration.
The slope must be 54-60 mV per decade of sulfide concentration. If the
slope is not acceptable, the ISE may not be working properly. For
corrective action, consult the ISE operating manual.
7.5 Allow samples and standards to equilibrate to room temperature prior
to analysis by ISE.
7.6 Prior to and between analyses, rinse the electrodes thoroughly with
reagent water and gently shake off excess water. Low-level measurements are
faster if the electrode tips are first immersed for five minutes in reagent
water.
7.7 Measurement of Sulfide in Distilled Samples
7.7.1 Pour 25.0 ml of sample into a 50 ml beaker. Add a Teflon®-
coated magnetic stir bar. Place the beaker on a magnetic stir plate and
stir at a slow speed (no visible vortex). Immerse the electrode tips to
just above the rotating stir bar. Record the meter reading (mV or
concentration) as soon as the reading is stable, but in no case should the
time exceed five minutes after immersing the electrode tips. If reading
mV, determine sulfide concentration from the calibration curve.
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7.8 When analyses have been completed, rinse the electrodes thoroughly
and store them in a 1.0 mg/L sulfide standard solution. If the electrodes will
not be used more than one day, drain the reference electrode internal filling
solutions, rinse with reagent water, and store dry.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and the distillation methods for specific
quality control procedures.
8.2 Initial Calibration Verification standard (ICV): After performing
the calibration step, verify calibration by analyzing an ICV. The ICV contains
a known sulfide concentration at the mid-range of the calibration standards and
is from an independent source. ICV recovery must be 90-110 percent. If not, the
error source must be found and corrected. An acceptable ICV must be analyzed
prior to sample analysis. The ICV also serves as a laboratory control sample.
8.3 Continuing Calibration Verification standard (CCV): After every 10
samples, and after the final sample, a CCV must be analyzed. The CCV contains
a known sulfide concentration at mid-calibration range. CCV recovery must be 90-
110 percent. If not, the error source must be found and corrected. If the ISE
calibration has changed, all samples analyzed since the last acceptable CCV must
be re-analyzed.
8.4 Reagent blank: After the ICV and after every CCV, a reagent blank
must be analyzed. A reagent blank is reagent water mixed 100:1 with SAOB. The
indicated reagent blank concentration must be less than 0.05 mg/L sulfide. If
not, the contamination source must be found and corrected. All samples analyzed
since the last acceptable reagent blank must be re-analyzed.
8.5 Matrix spike: Follow the matrix spike protocols presented in Chapter
One. The spike concentration must be 10 times the detection limit and the volume
added must be negligible (less than or equal to one-thousandth the sample aliquot
volume). Spike recovery must be 75-125 percent. If not, samples must be
analyzed by the method of standard additions.
8.6 The sulfide calibration standards may degrade by more than 10% from
day-to-day. The standards must be standardized daily before use (by titration)
and checked throughout the day if used as QC samples.
9.0 METHOD PERFORMANCE
9.1 The sulfide ISE was calibrated with 100 and 1,000 mg/L standards, and
a series of sulfide standards was analyzed as unknowns. The results are listed
in Table 2. As shown, recoveries ranged from 76-124% over the range 0.25-12,000
mg/L sulfide. This indicates that there is no practical difference between the
true and observed values for sulfide over this concentration range.
9.2 Three acid-soluble sulfide samples were prepared (low, medium, and
high; 25, 100, and 1,000 mg/L). Triplicate measurements of each were performed
in the following order: medium, low, high, high, low, medium. The data are
provided in Table 3.
9.3 Precision estimates were calculated from the average percent RSD
9215 - 5 Revision 0
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taken within each block of triplicate measurements in Table 3. The CV% was
calculated as 2.0 to 10.
9.4 Accuracy estimates from average percent recovery reported in Table
3 were calculated to be 75-105%.
9.5 To test the efficiency of the SAOB as a scrubber solution, three
standards (1, 10, and 40 mg/L) were distilled into SAOB scrubber and the
resulting sulfide concentration measured. The results are listed in Table 4.
As seen in the table, excellent recoveries are obtained using the SAOB scrubber
solution. The one low recovery for the 40 mg/L standard is most likely due to
incomplete sparging of oxygen from the system prior to distillation.
10.0 REFERENCES
1. Franson, Mary Ann H., Ed. Standard Methods for the Examination of
Water and Wastewater, 18th Edition. American Public Health Association,
Washington, DC, 1992.
2. Model 94-16 Silver/Sulfide Electrode Instruction Manual. Orion
Research, Inc., Boston, MA, 1986.
3. Hillerman, D.C., Nowinski, P. "Modification of Methods 9030 and 9031
for the Analysis of Sulfide by Specific Ion Electrode". U.S. Environmental
Protection Agency, EMSL-LV. EPA/600/4-90/024. September 1990.
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Table 1. Interference Study Results
Interferant
Ag+ (20 mg/L)
Ag+ + NH4+ (20 mg/L
each)
Hg+ (20 mg/L)
Humic acid (100 mg/L)
Humic acid + sulfide
(100 mg/L each)
Sulfide Concentration (mg/L)
DI matrix
<1
<1
<1
2.1
114
Tap water matrix
<1
<1
<1
1.6
87,8
* The humic acid contains 2-3 mg/L sulfide as determined by Method 9030.
Table 2. Results From a Single-Laboratory Accuracy
Evaluation of a Sulfide ISE
Sulfide Sulfide
Concentration Detected
(mg/L) (mg/L)
0.10
0.25
0.50
1.00
1.09
5.00
27.4
55.0
110
591
1,183
6,000
12,000
0.01
0.19
0.47
0.99
1.10
4.94
26.8
52.3
109
607
1,157
6,028
14,850
Sulfide
Recovery
(percent)
10
76
94
99
101
99
98
95
99
103
98
100
124
Rel. Std.
Deviation
(percent)
-
12.7
9.3
.
3.4
-
-
-
-
0.4
2.0
2.2
9215 - 7
Revision 0
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i
Table 3. Precision and Accuracy Data for the Sulfide ISE
True
(mg/L)
23.6
118.1
1,503
Measured
(mg/L)
19.2
114.8
1,455
Std. Dev.
(mg/L)
0.81
2.2
26
RSD (%)
4.2
1.9
1.8
Recovery
(%)
82.5
97.2
96.8
n = 6 for all samples
Table 4. Recovery of Hydrogen Sulfide in SAOB Solution
Sulfide (mg/L)
1
10
40
% Recovery
91.0
89.8
86.7
96.6
100
96.0
69.3
98.9
89.2
9215 - 8
Revision 0
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METHOD 9215
POTENTIOMETRIC DETERMINATION OF SULFIDE
IN DISTILLED SAMPLES WITH ION-SELECTIVE ELECTRODE
r
7.1 Add 10 mL of SAOB
solution and 40 mL of
reagent water; proceed
to distill sample.
>
r
7.2 Standardize
silver nitrate.
>
r
7.3 Standardize sulfide
standards.
1
r
7.4 Calibrate sulfide ISE.
>
f
7.5 Allow samples &
standards to equilibrate
to room temperature.
>
r
7.6 Rinse electrodes.
>
r
7.7 Take measurement &
determine concentration.
>
r
7.8 Drain reference
electrode and rinse.
^
r
9215 - 9
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CHAPTER SIX
PROPERTIES
The following methods are found in Chapter Six:
Method
Method
Method
Method
Method
Method
Method
Method
Method
1030:
1120:
1312:
1320:
1330A:
9041A:
9045C:
9050A:
9080:
Method 9081:
Method 9090A:
Method 9095A:
Method 9096:
Appendix A:
Method 9100:
Method 9310:
Method 9315:
Ignitability of Solids
Dermal Corrosion
Synthetic Precipitation Leaching Procedure
Multiple Extraction Procedure
Extraction Procedure for Oily Wastes
pH Paper Method
Soil and Waste pH
Specific Conductance
Cation-Exchange Capacity of Soils (Ammonium
Acetate)
Cation-Exchange Capacity of Soils (Sodium Acetate)
Compatibility Test for Wastes and Membrane Liners
Paint Filter Liquids Test
Liquid Release Test (LRT) Procedure
Liquid Release Test Pre-Test
Saturated .Hydraulic Conductivity, Saturated
Leachate Conductivity, and Intrinsic Permeability
Gross Alpha and Gross Beta
Alpha-Emitting Radium Isotopes
SIX - 1
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METHOD 1030
IGNITABILITY OF SOLIDS
1.0 SCOPE AND APPLICATION
1.1 This method is suitable for the determination of the ignitability of
solids and is appropriate for pastes, granular materials, solids that can be cut
into strips, and powdery substances. This method may be used to meet certain
regulatory applications but is not required for determining if a waste passes or
fails the characteristic of ignitability per the RCRA definition. If it is
impractical to perform the test because of the physical form of the sample,
generator knowledge should be used to determine the ignitability hazard posed by
the material.
2.0 SUMMARY OF METHOD
2.1 In a preliminary test, the test material is formed into an unbroken
strip or powder train 250 mm in length. An ignition source is applied to one end
of the test material to determine whether combustion will propagate along 200 mm
of the strip within a specified time period. Materials that propagate burning
along a 200 mm strip within the specified time period are then subjected to a
burning rate test. Materials that do not ignite or propagate combustion as
described above do not require further testing. In the burning rate test, the
burning time is measured over a distance of 100 mm and the rate of burning is
determined. The test method described here is based on the test procedure adopted
by the Department of Transportation from the United Nations regulations for the
international transportation of dangerous goods and is contained in Appendix E
to Part 173 of 49 CFR.
3.0 INTERFERENCES
3.1 In laboratory tests the burning rate of duplicate runs is usually
repeatable to within 10%. However, large differences in burning rates may occur
if experimental conditions are not held constant. Variation in airflow rates,
particle size, and moisture content of the test material will affect test
results. Therefore, at least triplicate determinations of the burning rate
should be conducted.
3.2 Particle size of test material can affect not only the burning rate,
but also the ignition of the material. Therefore, the particle size of the test
material should be the same for each test run. The particle size of the test
material should be reported in a simple descriptive format (e.g., fine powder,
sand, coarse granular).
3.3 Temperature of some test material such as sulfur powder affects the
burning rate. For reproducible results, all tests should be performed at
approximately the same initial temperature (ambient room or laboratory
temperature).
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3.4 All tests must be carried out inside a fume hood with the test
apparatus situated perpendicular (90°) to the direction of airflow. Airflow
parallel (0°) to the test apparatus results in non-reproducible burning rates.
3.5 The rate of airflow through the fume hood affects the burning rate.
Too high an airflow distorts the flame and retards its horizontal propagation.
The optimum airflow appears to be in the range of 0.7-1 meter per second.
3.6 Materials that are moisture sensitive (i.e., readily absorb moisture
from air) should be tested as quickly as possible after removal from the sample
container. All materials should be tested as received by the laboratory.
4.0 APPARATUS AND MATERIALS
4.1 Low-heat conducting, non-combustible, impervious ceramic tile or
equivalent material, of approximate dimension of 25 cm x 25 cm x 2.5 cm (the tile
must be at least 25 cm in length to support a 250 mm test sample).
4.2 High temperature marker or equivalent making device for marking
ceramic plates.
4.3 Powder Train Mold (see Figure 1) for molding powdered and granular
materials for the burn rate test. The material of construction can be aluminum,
brass, or stainless steel. The mold is 250 mm in length and has a triangular
cross-section, with a width of 20 mm, and a depth of 10 mm as measured from the
bottom of the triangular opening to where the sides meet. On both sides of the
mold, in the longitudinal direction, two metal sheets are mounted as lateral
limitations which extend 2 mm beyond the upper edge of the triangular cross-
section. This device can be fabricated by most machine shops. The complete burn
rate apparatus is available from: Associated Design and Manufacturing Co.; 814
N. Henry Street; Alexandria, Virginia 22314.
4.4 A Bunsen (propane gas and air) burner with a minimum diameter of 5
mm capable of attaining a temperature of at least 1,000°C.
4.5 Stop watch
4.6 Thermocouple to measure the temperature of the gas flame.
4.7 Thermometer to measure initial temperature of material (i.e., room
temperature).
4.8 Anemometer to measure airflow in the fume hood.
5.0 REAGENTS
5.1 No special reagents are required to conduct this test.
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6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples are tested on as-received basis unless requested
otherwise. No sample preservation is required, but sample containers should be
completely filled and tightly sealed to preserve sample integrity.
6.2 Samples should be tested as soon as possible after removal from the
sample container (i.e., samples should not be allowed to dry or absorb moisture
for excessive periods or to loose volatiles). Samples that are chilled or cooled
upon receipt to the laboratory should be allowed to equilibrate to the ambient
laboratory temperature in the sample container.
7.0 PROCEDURE
SAFETY; Prior to starting the preliminary test, all sample materials
must be tested to determine if that material is explosive or
extremely flammable. Use a very small portion of material (1 gram
or less). If the sample displays explosivity or extreme
flammability, do not conduct this test.
7.1 Preliminary Screening Test
7.1.1 The preliminary ignitability test is conducted on all waste
materials. On a clean, impervious ceramic tile (Section 4.1), clearly
mark a 250 mm long test path. Make another mark at exactly 200 mm from
the start of the sample path.
7.1.2 Prepare the test material in its "as received" form by
forming an unbroken strip or powder train of sample 250 mm long by 20 mm
wide by 10 mm high on the ceramic tile. Use the mold to form the material
as in 7.2.3 if appropriate.
7.1.3 Place the ceramic tile with the loaded sample in a fume hood
about 20 cm (~8 inches) from the front of the hood and in an area of
laminar airflow. Position the sample perpendicular to the airflow. (See
Figure 2) The airflow across the perpendicular axis of the sample should
sufficient to prevent fumes from escaping into the laboratory and should
not be varied during the test. The air velocity should be approximately
0.7 meters/second. Measure the air velocity by an anemometer.
7.1.4 Light the Bunsen burner and adjust the height of the flame
(6.5 to 7.5 cm) by adjusting the propane gas and air flows. Measure the
temperature of the flame (tip of the flame) by a thermocouple. The
temperature of the flame must be at least 1000°C.
7.1.5 Apply the tip of the flame to one end of the sample strip .
The test period will depend on the sample matrix as follows:
7.1.6 If the waste is non-metallic, hold the flame tip on the
sample strip until the sample ignites or for a maximum of 2 minutes. If
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combustion occurs, begin timing with a stop watch and note whether the
combustion propagates up to the 200 mm mark within the 2 minute test
period.
7.1.7 If the waste is a metal or metal-alloy powder, hold the flame
tip on the sample strip until the sample ignites or for a maximum of 5
minutes. If combustion occurs, begin timing with a stop watch and note
whether the combustion propagates up to the 200 mm mark within the 20
minute test period.
7.1.8 If the waste does not ignite and propagate combustion either
by burning with open flame or by smoldering along 200 mm of sample strip
within the 2 minute test period (or 20 minute test period for metal
powders), the waste is not considered flammable and no further testing is
required. If the waste propagates burning of 200 mm of the test strip
within the 2 minute test period (20 minute test period for metals), the
material must be evaluated by the burning rate test (Section 7.2).
7.2 Burning Rate Test
7.2.1 The preparation of the test sample for the burning rate test
will depend on the physical characteristics of the waste. Wastes that
exist in a powdered or granular state are molded in a powder train mold
shown in Figure 1. Pasty materials are formed into a rope 250 mm in length
with a cross-section of 1 cm2. All tests for the burn rate test are
performed on clean, ambient temperature, ceramic plates.
7.2.2 On a clean, impervious ceramic tile (Section 4.1), clearly
mark a 250 mm long test path. Make two additional timing marks at 80 mm
and 180 mm from the start of the sample path. The distance between the
two marks (100 mm) will be used to calculate the rate of burn in Section
7.2.9.
7.2.3 Tighten the side plates on the mold. For powdered or granular
materials: Place the mold on the base plate. Pour the material to fill
the triangular cross section of the mold loosely.
7.2.4 Drop the unit from a height of 2 cm onto a solid surface
three times to settle the powder. Remove the side supports. Lift the
mold off the base plate. Place a clean ceramic test plate with the
appropriate timing marks (Section 7.2.2) face down on top of the mold.
Invert the setup and remove the mold.
7.2.5 Pasty wastes are prepared by spreading the waste on a marked
ceramic tile (Section 7.2.2) in the form of a rope 250 mm in length with
a cross-section of 1 cm2.
7.2.6 Place the ceramic tile with the loaded sample prepared in
Sections 7.2.3 or 7.2.5 in a fume hood about 20 cm (~8 inches) from the
front of the hood and in an area of laminar airflow. Position the sample
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perpendicular to the airflow. (See Figure 2) The airflow across the
perpendicular axis of the sample should sufficient to prevent fumes from
escaping into the laboratory and should not be varied during the test.
The air velocity should be approximately 0.7 meters/second. Measure the
air velocity with an anemometer.
7.2.7 Light the Bunsen burner and adjust the height of the flame
(6.5 to 7.5 cm) by adjusting the propane gas and air flows. Measure the
temperature of the flame (tip of the flame) by a thermocouple. The
temperature of the flame must be at least 1000°C.
7.2.8 Apply the tip of the flame to one end of the sample strip to
ignite the test strip as described in Section 7.1.6 and 7.1.7.
7.2.9 When the test strip or powder train has burned up to the 80
mm time marker, begin timing the rate of combustion with a stop watch.
Stop the timer when the burned strip reaches the 180 mm time marker.
Record the amount of time (in seconds) required to burn the 100 mm test
strip. Calculate the rate of burning by dividing the length of the burn
test strip (100 mm) by the total time (seconds). Results of the burn rate
test should be reported in mm/sec. Wastes that have a rate of burning of
more than 2.2 mm/sec (or burn time of less than 45 seconds for 100 mm) are
considered to have a positive result for ignitability according to DOT
regulations. For metals, this time is 10 minutes or less for 100 mm (or
a burn rate of more than 0.17 mm/sec).
7.2.10 Report and Calculation Section
Test Material Information
Source of Material: e.g., Company, operation or process
Description of material: e.g., powder or paste, metallic or non-metallic
Particle size: e.g., fine powder, granular, sand, etc.
Preliminary Burning Time: seconds.
Test Conditions
Date of Test:
Temperature of test material (°C):
Air velocity through fume hood (m/s):
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Ignitability Test Data
Test
Number
1
2
3
Time (sec)
elapsed between
application of
flame and start
of ignition
Burning
time over
100 mm
(sec)
Burning
Rate
(mm/sec)
Comments
8.0 QUALITY CONTROL
8.1 All tests must be performed on a clean ceramic plate at room
temperature. All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
8.2 All replicate runs must be at the same initial temperature (ambient
laboratory temperature).
8.3 All replicate tests must be run at approximately the same airflow
through the fume hood.
8.4 Only materials of the same particle size distribution should be used
for all replicate tests.
8.5 The burn rate test must be conducted in triplicate if the preliminary
screening test is positive. Any burn rate for non-metallic samples that exceeds
2.2 mm/sec (or a burn time of less than 45 seconds for 100 mm) is considered to
have a positive result. For metals, a burn rate of more than 0.17 mm/sec (or
burn time of less than 10 minutes for 100 mm) is considered to have a positive
result.
9.0 METHOD PERFORMANCE
9.1 An independent laboratory validation was conducted on the robustness
of the burn rate test procedure. The materials selected for this evaluation
included:
1. A 50/50 mixture of metallic silicon and lead dioxide (PB02)
2. Excelsior
3. Dextrin (yellow powder)
4. Sulfur (fine yellow powder)
5. Aluminum metal (coarse)
6. Magnesium metal (coarse)
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7. Polyethylene high density (granular)
8. Polyethylene low density (fluffy white powder)
9. Scott fertilizer (32-3-10:N-P-K)
10. JP-4 contaminated soil (approximately 5000 ppm)
Of these materials, the 50/50 mixture of metallic silicon and lead dioxide
(PB02), elemental sulfur, and excelsior were considered to give a positive
ignitability result under the conditions of the test. The remaining materials
gave negative (nonflammable) results under the conditions of the test. Several
test variables including ignition source, ambient temperature, and apparatus
orientation, were studied using these materials. Partial results of this study
are summarized in Table 1.
Table 1 Test Variables for Ignitability
Material
Tested
50% Metallic
Silicon and 50%
Lead IV Oxide
Excelsior
(wood shavings)
Test
Number
1
2
3
4
1
2
3
4
Variable
combination1
ABC
Abe
aBc
abC
ABC
Abe
aBc
abC
Burn Time
over 100 mm
(sec)
0.84
0.50
0.69
0.65
13.45
9.14
13.37
13.59
Burn Rate
(mm/sec)
119
200
145
154
7.43
10.9
7.47
7.36
1where:
A-flame ignition
a-hot wire ignition source
B-ambient temperature of 20°C
b-ambient temperature of 100'C
C-orientation of test apparatus of 90* to air flow
c-orientation of test apparatus of 0° to air flow
9.2 In another evaluation of the DOT burn rate test, potentially ignitable
finishing wastes from the furniture industry were collected and tested for
burning rates. Each waste was tested in triplicate to establish a mean value
for the burning rate. The results for the flammable wastes are summarized in
Table 2.
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Table 2-Burning Rates for Ignitable Wastes
Sample No.
A2
J2
U
K
H
F
P
0
Q
Description of Waste
Segregated Lacquer Dust
Segregated Lacquer Dust
Segregated Lacquer Dust
Consolidated Lacquer Dust
Catalyzed Lacquer Dust
Water Based Lacquer Dust
Booth Coat-Stain Overspray
Pallet Covered Cardboard
Pallet Covered Cardboard
Burn Time over 100
mm (sec)
4.7
4.6
8.6
6.0
6.7
19.4
12.5
11.1
12.3
Burn
Rate
(mm/sec)
21.3
21.7
11.6
16.7
14.9
5.15
8.0
9.0
8.13
9.3 In order to evaluate the ruggedness of the DOT burn rate test, select
ignitable finishing wastes were split and tested by a state laboratory and an
independent contract laboratory. The results of this comparison are summarized
in Table 3.
Table 3-Comparison of Burn Rates
Sample No.
Al
Jl
12
Description of
Waste
Segregated Lacquer
Dust
Segregated Lacquer
Dust
Booth Coat-Glaze
Overspray
Mean Burn Time Over 100 mm in
Seconds
State
Laboratory
4.7
4.6
O1
Contract
Laboratory
5
4.3
O1
1waste was found to be nonflammable under conditions of the test.
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10.0 REFERENCES
1. "Test Methods for Readily Combustible Solids. Burning Rate
Test." (14.2.2.5). Recommendations on the Transport of
Dangerous Goods. Fifth Revised Edition. United Nations, New
York. 1988.
2. DOT Regulation. Appendix E to Part 173 of 49 CFR, Chapter 1
(12-31-91 Edition), pp. 597-598.
3. Flammability (solids). Method A.10. Official Journal of the
European Communities. 9/19/84. No. L251/63.
4. "Validation of Ignitability Method For Solids" Foster Wheeler
Enviresponse, Inc., Edison NJ., Submitted to the Office of
Solid Waste, US EPA, February 1994.
5. Internal Report, (AMFA Report) North Carolina Department of
Environmental Health and Natural Resources. (Bill Hamner)
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Figure 1
Powder Train Mold
(A) Cross-section of 250 mm long mould
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Figure 2
Test Apparatus Position in Fume Hood
T
8"
1
i M
t
a- '
AIR FLOW
HOOD SASH °-7 m/s
SAMPLE 90* TO AIR FLOW
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METHOD 1030
IGNITABILITY OF SOLIDS
i
Nonmetallio
7.1.6 Apply flame to
test strip, hold
for 2 minutes.
>
1
7.1 1 Perform preliminary
screening test.
^
r
71.1-7.12 Mark
ceramic plate,
prepare test strip
or powder train.
1
r
7.1 3 Position apparatus
in fume hood 90°
to air flow.
^
r
7 1 4 Light flame and
measure temperature.
Metallic
7.1.7 Apply flame to
test strip, hold
for 5 minutes.
Nonflammable Solid
No
Nonflammable Solid
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METHOD 1030
TGNITABILITV OF SOLIDS
7 2.3 - 7.2.4 Pour waste
into the mold, drop
three times, invert
on marked ceramic
plate.
7 2.5 Spread pasty
waste on marked
ceramic plate,
250 mm x 1 cm2.
7.2.6 Position test sample
in a fume hood 90 to
atr flow.
7.2 7 Light bunsen
burner, adj. flame and
measure temperature.
7.2 8 Apply flame to end
of the strip to ignite.
7.2.9 Note time (seconds)
needed for the flame to
travel 100 mm distance.
Calculate burn rate.
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METHOD 1120
DERMAL CORROSION
1.0 SCOPE AND APPLICATION
1.1 The dermal corrosion assay system is an in vitro test method which
determines the corrosive potential of a substance toward human skin. The method
is simple, rapid, accurate, and may be applied to both solids, liquids and
emulsions. The liquids may be aqueous or non-aqueous. Solids can be water-
soluble or non-soluble. The samples may be pure chemicals, dilutions,
formulations, or waste. No prior treatment of the sample is required. This
method may be used to meet certain regulatory applications but is not required
for determining if a waste passes or fails the characteristic of corrosivit.y per
the RCRA definition.
2.0 SUMMARY OF METHOD
2.1 The assay system is an in vitro test method which is composed of two
components, a synthetic macromolecular biobarrier and a Chemical Detection System
(CDS). Test samples are applied on top of the macromolecular biobarrier.
Corrosive samples are able to disrupt the macromolecular structure of the
biobarrier. A color change in the CDS, located beneath the biobarrier, is
detected visually and indicates that the test sample has altered the biobarrier
sufficiently to allow its passage through the full thickness of the biobarrier.
The time it takes a sample to disrupt the biobarrier is inversely proportional
to the degree of corrosivity of the sample - the longer it takes to observe a
color change, the less corrosive the substance is. Noncorrosive samples do not
disrupt the biobarrier, or disrupt the biobarrier after a predetermined time
period (see Section 2.4).
Corrosive samples may be placed into three different classes of
corrosivity, established by the time required for the sample to break through the
biobarrier. These classes are called Packing Groups by the U.S. Department of
Transportation (DOT). Packing Groups are assigned according to the degree of
danger presented by the corrosive material; Packing Group I indicates great
danger; Packing Group II, medium danger; Packing Group III, minor danger. For
consistency, these same definitions are used for this test method and are
referred to as Group I, Group II, and Group III.
2.2 Prior to performing the assay, samples are pre-qualified to establish
their compatibility with the assay system. The sample is placed in a small
amount of CDS fluid. If any detectable change occurs in the CDS, the sample is
qualified and can be analyzed by the test. If a sample is non-qualified, it is
incompatible with the CDS and must be tested by another method.
2.3 Test samples are classified into categories by the screening test
which is supplied with the assay kit. The category that a sample is assigned to
will determine how the Groups will be assigned. Test samples are classified by
pH changes produced in two well-defined buffers - one designed to buffer acids
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and another that buffers bases. These buffers are supplied as part of the
screening test. Four different categories are defined as follows:
2.3.1 Category AT substances produce a large change in pH when they
are added to the acid buffer. This change in pH is indicated by a strong
color change of the acid buffer solution.
2.3.2 Category B, substances produce a large change in pH when they
are added to the base buffer. This change in pH is indicated by a strong
color change of the base buffer solution.
2.3.3 Category A2 substances produce little or no pH changes when
added to the acid buffer, and therefore, little or no color change in the
buffer solution is observed.
2.3.4 Category B2 substances produce little or no pH changes when
added to the base buffer, and therefore, little or no color change in the
buffer solution is observed.
2.4 Groups are assigned in the assay system by taking into account the
category that is assigned to a sample by the screening test, and the time it
takes to detect a color change in the CDS in the assay. Category ^ and B,
samples are assigned to Group I if a color change is observed between zero and
three minutes, to Group II if a color change is observed after three minutes and
up to one hour, and to Group III if a color change is observed after one hour and
up to four hours. If no color change occurs in four hours, the chemical is
classified as Noncorrosive.
Category A2 and B2 samples are assigned to Group I if a color change
is observed between zero and three minutes, to Group II if a color change is
observed after three minutes and up to 30 minutes, and to Group III if a color
change is observed after 30 minutes and up to 45 minutes. If no color change
occurs in 45 minutes, the chemical is classified as Noncorrosive.
3.0 INTERFERENCES
3.1 The test is not subject to interference from color, turbidity,
colloidal matter or high salinity.
3.2 The Pre-qualification Test, the Screening Test and the Assay must be
performed at room temperature. The samples must also be at room temperature (17-
25°C).
4.0 APPARATUS AND MATERIALS
4.1 Corrositex Assay Kit (InVitro International, 16632 Millikan Avenue,
Irvine, CA 92714). The following three items are supplied in the Corrositex
Assay Kit:
4.1.1 Four racks holding seven vials with black caps.
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4.1.2 One tray of 24 membrane discs.
4.1.3 Four data sheets (color charts).
4.2 Combination hot plate/stir plate or equivalent - able to heat to 75°C.
Stirring speed should be adjustable.
4.3 Digital thermometer - able to read to 75°C.
4.4 Timers (6) - able to measure hours, minutes and seconds.
4.5 Repeat pipettor - this pipet is different than the pipet specified in
Section 4.12. Delivers 200 /*L repeatedly, without refilling between individual
deliveries.
4.6 2.5 ml combitip for repeat pipettor.
4.7 Lab Industries or equivalent sample pipettor - a positive displacement
pipettor useful when pipetting viscous samples.
4.8 Pipet tips for Lab Industries, or equivalent, pipettor.
4.9 Test tubes
4.10 Balance - capable of weighing 100 mg accurately.
4.11 Spatula - capable of transferring 0.1 - 0.5 g.
4.12 Pipets - microliter, with disposable tips. Should be able to measure
100 L accurately.
4.13 Tweezers.
4.14 Permanent marker pens.
4.15 Plastic wrap.
5.0 REAGENTS
5.1 All reagents listed below are provided in the Corrositex Assay Kit
except for the positive and negative controls mentioned in Section 5.7. The
Corrositex Assay Kit is available from InVitro International, 16632 Millikan
Avenue, Irvine, CA 92714.
5.2 Chemical Detection System (CDS).
5.3 Screening test buffer solutions.
5.4 Confirmation Test Solution.
5.5 One gram of the biobarrier matrix and a microstir bar.
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5.6 10 ml of biobarrier diluent.
5.7 Positive and negative controls, if desired, for GLP purposes.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Appropriate precautions should be taken for handling potentially
corrosive substances such as wearing gloves and having proper eye protection.
6.2 Samples should be analyzed as soon as possible after collection.
7.0 PROCEDURE
7.1 Follow the established laboratory procedures for working with
hazardous test samples. Wear lab coat, gloves and safety glasses when working
with any potentially corrosive material.
7.2 Pre-Qualification Test
7.2.1 Add 100 mg or 150 pi of sample to 1.0 mL of CDS in duplicate
test tubes.
7.2.1.1 Sample qualifies if there is a color reaction within
5 minutes: proceed with assay.
7.2.1.2 If no reaction is observed, the sample is non-
qualified. Seek other methods to determine corrosivity.
7.3 Screening Test
7.3.1 Liquid samples
7.3.1.1 Add 150 L of sample to Test Tubes 1 and 2. Cap the
test tubes and shake vigorously for 10 seconds. Read the color
change of the mixture within one minute. If the sample is immiscible
in the solution, wait one minute and then read the color change at
the interface.
7.3.1.2 Assign the category. If an intense color change
(similar to the Category B, color chart) is observed in Tube 1,
assign the sample to Category B,. If an intense color change
(similar to the Category A, color chart) is observed in Tube 2,
assign the sample to Category A,. Proceed to the next step if no
intense color change is observed in Test Tubes 1 and 2.
7.3.1.3 Confirmation test. Add two drops of the Confirmation
Test Solution to Test Tube 1. Cap the test tube and shake vigorously
for 5 seconds. If the color of the solution changes to yellow or
gray (similar to the Category A2 color chart at the bottom of the
protocol sheet) the sample is classified as Category A2. If the
color of the solution changes to purple or blue (similar to the
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Category B2 color chart at the bottom of the protocol sheet) the
sample is classified as Category B2.
7.3.2 Solid samples
7.3.2.1 Add 100 mg of sample to Test Tubes 1 and 2. Cap the
test tubes and shake vigorously for one minute. Wait another minute
and read the color change of the mixture. If the sample is insoluble
in the solution, allow the mixture to settle and read the color
change at the interface of the solution and the solid.
7.3.2.2 Assign the category. If an intense color change
(similar to the Category B., color chart) is observed in Tube 1,
assign the sample to Category B,. If an intense color change
(similar to the Category AT color chart) is observed in Tube 2,
assign the sample to Category Av Proceed to the next step if no
intense color change is observed in Test Tubes 1 and 2.
7.3.2.3 Confirmation test. Add two drops of the Confirmation
Test Solution to Test Tube 1. Cap the test tube and shake vigorously
for 5 seconds. If the color of the solution changes to yellow or
gray (similar to the Category A2 color chart at the bottom of the
protocol sheet) the sample is classified as Category A2. If the
color of the solution changes to purple or blue (similar to the
Category B2 color chart at the bottom of the protocol sheet) the
sample is classified as Category B2.
7.4 Assay
7.4.1 Biobarrier preparation
7.4.1.1 Biobarrier matrix preparation - must be completed at
least two hours prior to running assay.
7.4.1.2 Place scintillation vial containing biobarrier matrix
powder on the hot plate pad. Begin spinning the stir bar before
adding the diluent.
7.4.1.3 Add the entire contents of the biobarrier diluent
vial slowly and constantly to the vial of biobarrier matrix powder.
Make sure that the stir bar is turning while adding the diluent. The
stir bar should be turning rapidly, but not so fast that the solution
foams.
7.4.1.4 Turn the heat on low; monitor the temperature of the
solution as it is warming. Gradually increase the heat as necessary
to warm the solution to 68°C (±1°C) to solubilize the matrix. This
may take approximately 20 minutes. DO NOT allow the temperature to
exceed 70°C.
7.4.1.5 While the solution is warming, remove the tray of 24
membrane discs from the refrigerator. Remove the tray lid.
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7.4.1.6 After the biobarrier matrix solution reaches 68°C and
has been completely solubilized, turn off the heat and move the vial
toward the edge of the heat pad to keep it warm while aliquotting
into the membrane discs.
7.4.1.7 Dispense solubilized solution into membrane discs.
Using the repeat pipettor, set to dispense 200 /xL. Slowly fill the
pipette tip with biobarrier solution, avoiding air bubbles. Dispense
one aliquot back into the biobarrier vial to ensure proper subsequent
volume delivery. Wipe the tip before dispensing each aliquot.
Dispense 200 fj.1 into each disc, ensuring that the entire membrane is
covered and no air bubbles have formed. Any air bubbles in the gel
will alter the results of the test, therefore the disc cannot be
used.
7.4.1.8 Label the lid with the date, time, lot number, and
initials of the technician preparing the biobarrier. Wrap the filled
tray evenly with plastic wrap; do not bunch up the plastic wrap
underneath the plate.
7.4.1.9 Store the tray at 2 - 8°C for at least two hours
before beginning any testing. The biobarrier is stable for fourteen
days if wrapped and stored at 2 - 8°C.
7.4.2 Running the assay.
7.4.2.1 On the data sheet, complete the lot number, date,
time, name of technician, name of chemical, whether it is solid or
liquid, and pH of a 10% solution diluted in water or appropriate
solvent.
7.4.2.2 When ready to test, remove one tray of seven black-
capped scintillation vials from the kit box.
7.4.2.3 Using a pipet-aid, dispense 22.0 mL of the Chemical
Detection System (CDS) into each of the six (6) scintillation vials.
Make sure that the CDS is at room temperature (17 - 25°C) before
using. Dispense 12.0 ml of the CDS into the seventh scintillation
vial and cap it. This vial will serve as a color control.
7.4.2.4 Remove the tray of 24 membrane discs from the
refrigerator. Place on a tray of ice.
7.4.2.5 Place disc into first scintillation vial. Do not
allow the discs to be in contact with the CDS for longer than two
minutes before applying the test sample. Within two minutes, add 500
pi (using the Lab Industries pipettor, or equivalent) or 500 mg
(using spatula or tweezers) of test sample to disc. Start timer the
instant the sample is added.
Note: Do not cap the vials while test is in progress due to
potential pressure build-up.
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7.4.2.6 Watch the vial for three minutes, ensuring that the
color reaction is not missed if it is a Group I sample. Changes in
the CDS may include various color changes, flaking or precipitation.
7.4.2.7 Add three more discs and samples to vials, staggering
each start time so that the most accurate reaction times are
recorded.
7.4.2.8 Allow assay to run until color or physical reaction
occurs. Category A., and ft, samples should be checked for reactions
at 3 minutes, 1 hour and 4 hours. Category A2 and B2 samples should
be checked for reactions at 3, 30 and 45 minutes.
7.4.2.9 At the first indication of the presence of a chemical
reaction in the CDS, there will be a color change produced beneath
the bottom-center of each biobarrier disc. As soon as a reaction is
observed, immediately record net time of each vial on the data sheet.
7.4.2.10 Run positive and negative controls in the other two
vials, if desired, for GLP purposes.
7.4.3 Assignment of Groups.
7.4.3.1 Category A, and B., samples are assigned to Group I if
a color change is observed between zero and three minutes, to Group
II if a color change is observed after three minutes and up to one
hour, and to Group III if a color change is observed after one hour
and up to four hours. If no color change occurs in four hours, the
chemical is classified as Noncorrosive.
7.4.3.2 Category A2 and B2 samples are assigned to Group I if
a color change is observed between zero and three minutes, to Group
II if a color change is observed after three minutes and up to 30
minutes, and to Group III if a color change is observed after 30
minutes and up to 45 minutes. If no color change occurs in 45
minutes, the chemical is classified as Noncorrosive.
8.0 QUALITY CONTROL
8.1 Samples should be analyzed in quadruplicate. The test may be analyzed
in duplicate if a simple screening of corrosives and noncorrosives is all that
is required. However, it is recommended that for greater accuracy, samples be
analyzed in quadruplicate. It is suggested that positive and negative controls
be analyzed to conform with GLP.
9.0 METHOD PERFORMANCE
9.1 Interlaboratory and intralaboratory studies were conducted with five
different laboratories. Ten different chemicals were tested with six replicates.
The data are presented in Table 1.
1120 - 7 Revision 0
January 1995
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9.2 More than 200 data points have been collected at InVitro International
for six reference samples. Statistical analysis of this data shows the standard
deviation for ethylenediamine and ferric chloride is about 5% of their respective
assay times, and about 10% for maleic anhydride, sodium hydroxide, and
dicyclohexylamine. The standard deviation for sulfuric acid approaches 18% of
its assay time, but when taken into account that the mean assay time is less than
1 minute, a standard deviation of 0.13 minutes is actually a reflection of the
difficulty of measuring such brief timeperiods.
9.3 The Corrositex assay has been used by more than 300 laboratories to
test approximately 4,000 test materials in its first phase of utilization in
industry. Diverse chemicals and formulations which include liquids, solids,
insolubles and immiscibles have been studied from many major industries including
petrochemical, agrochemical, surfactant, textile, paper and pulp, electroplating
and water treatment. Examples of dermal corrosion values compaired to pH for
selected compounds are shown in Table 2.
9.4 Data results from 1,050 samples that have been tested using the assay
system were complied and compared with in vivo data. Ninety-two percent of the
samples (965 samples) passed the Pre-qualification Test and were then analyzed
in the screening test and the assay. Assay was found to be highly concordant
with corrosive/noncorrosive in vivo results. Of 406 corrosive samples with in
vivo data, 377 (93%) were correctly identified as corrosive by assay. Of 296
noncorrosive samples with in vivo data, 83% were identified as noncorrosives,
demonstrating the ability of this in vitro method to correctly identify
corrosives and noncorrosives. Assay was also shown to accurately predict Packing
Groups. Six hundred out of 702 samples (85.5%) were placed in the same Packing
Group as that indicated by in vivo testing. Only 38 test samples out of 702
samples that had in vivo data were found to underestimate (5.4%). Of these 38
samples, 28 were distinct samples and the remainder were samples that had been
tested in more than one laboratory. When taking this information into account,
the percent of underestimation decreased to about 4%.
10.0 REFERENCES
1. Code of Federal Regulations, Transportation Title 49, Hazardous Materials
Table, Section 172.101 (1991).
2. Code of Federal Regulations, Transportation Title 49, Method of Testing
Corrosion to the Skin, Part 173, Appendix A (1991).
3. Schlesselman, J.J. (1973) Planning a Longitudinal Study: I. Sample Size
Determination. J. Chron. Dis. 26, 553-560.
4. ASTM Standards on Precision and Bias for Various Applications, "Standard
Practice for Conducting an Interlaboratory Study to Determine the Precision of
a Test Method"; ASTM: Philadelphia, PA, 1992; E 691-92.
5. Gordon, V.C., Marvel 1, J., and Maibach, H. (1994). Dermal Corrosion, The
Corrositex System, A DOT Accepted Method to Predict Corrosivity of Test
Materials. In Vitro Toxicology. Ed. Mary Ann Liebert, 1994.
1120 - 8 Revision 0
January 1995
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TABLE 1
LABORATORY DATA
Dichloroacetyl chloride
Laboratory
Mean of 6 Test
Results (minutes)
Standard
Deviation
Relative Standard
Deviation
1
2
3
4
5
3.30
2.99
3.88
2.50
3.30
0.59
0.30
0.47
0.26
0.28
17.88%
10.03%
12.11%
10.40%
8.48%
Formic acid
Laboratory
Mean of 6 Test
Results (minutes)
Standard
Deviation
Relative Standard
Deviation
1
2
3
4
5
6.32
5.08
5.18
4.82
4.02
0.51
0.46
0.77
0.29
0.29
8.07%
9.06%
14.86%
6.02%
7.21%
Dichloroacetic acid
Laboratory
Mean of 6 Test
Results (minutes)
Standard
Deviation
Relative Standard
Deviation
1
2
3
4
5
6.92
5.21
6.32
5.78
5.65
0.32
0.25
0.98
0.26
0.46
4.62%
4.80%
15.51%
4 . 50%
8.14%
Chloroacetic acid
Laboratory
Mean of 6 Test
Results (minutes)
Standard
Deviation
Relative Standard
Deviation
1
2
3
4
5
5.46
7.83
4.95
6.91
4.95
0.36
0.00
0.35
0.94
0.34
6.59%
0 . 00%
7.07%
13.60%
6.87%
1120 - 9
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TABLE 1 (continued)
Dodecyltri chlorosllane
Mean of 6 Test
Laboratory Results (minutes)
Standard
Deviation
Relative Standard
Deviation
1
2
3
4
5
10.78
11.59
11.22
11.96
10.98
0.05
0.36
0.82
0.56
0.29
0.46%
3.11%
7.31%
4.68%
2.64%
Ammonium hydrogen sulfate
Laboratory
Mean of 6 Test
Results (minutes)
Standard
Deviation
Relative Standard
Deviation
1
2
3
4
5
10.47
9.02
13.82
11.17
7.88
0.56
0.33
1.0
0.93
0.26
5.35%
3.66%
7.24%
8.33%
3.30%
Ethylenediamine
Laboratory
Mean of 6 Test
Results (minutes)
Standard
Deviation
Relative Standard
Deviation
1
2
3
4
5
17.24
21.33
26.12
20.76
22.48
0.38
0.53
1.30
0.19
1.40
2.20%
2.48%
4.98%
0.92%
6.23%
Aluminum chloride
Laboratory
Mean of 6 Test
Results (minutes)
Standard
Deviation
Relative Standard
Deviation
1
2
3
4
5
11.91
21.33
26.12
20.76
22.48
0.41
0.53
1.30
0.19
1.40
3.44%
2 . 48%
4 . 98%
0.92%
6.23%
1120 - 10
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TABLE 1 (continued)
Acetic acid
Laboratory
Mean of 6 Test
Results (minutes)
Standard
Deviation
Relative Standard
Deviation
1
2
3
4
5
28.52
27.00
34.98
36.30
29.67
0.87
0.00
0.22
0.41
0.62
3.05%
0.00%
0.63%
1.13%
2.09%
Dicyclohexylamine
Laboratory
Mean of 6 Test
Results (minutes)
Standard
Deviation
Relative Standard
Deviation
1
2
3
4
5
181.73
168.83
210.70
159.04
126.75
0.47
9.11
7.68
7.58
0.62
0.26%
5.40%
3.64%
4.77%
0.49%
1120 - 11
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TABLE 2. EXAMPLES OF DERMAL CORROSION VALUES FOR SELECTED COMPOUNDS
Compound Name
Acetic acid
Aluminum chloride
Ammonium hydroxide
Bromoacetic acid
Butyl ami ne
Citric acid
1,2-Diaminopropane
Dichloroacetic acid
Dicyclohexylamine
Di ethyl ami ne
Diethylene triamine
Ethanolamine
Ferric chloride
Formic acid
Hydrochloric acid
Hexanoic acid
Maleic acid
Mercaptoacetic acid
Nitric acid
Phosphoric acid
Potassium hydroxide
Propionic acid
Sodium hydroxide
Sodium metasil icate
Sulfuric acid
Thiophosphoryl chloride
Tri butyl ami ne
Trichlorotoluene
Triethanolamine
Triphosphoryl chloride
Concentration
(weight %)
99+
pure
10.00
55.60
40.00
20.00
99+
3.10
99.00
98.00
99.00
99+
98.00
33.90
35.00
99.00
99.00
15.10
90.00
85.00
pellets
99+
pellets
20.00
15.00
98.00
99+
99.00
60.00
98.00
PH1
0.00
2.92
12.37
0.93
12.96
1.28
12.06
0.98
9.57
13.86
12.01
11.82
3.00
0.62
0.00
3.00
1.30
1.60
0.00
0.00
14.00
0.35
13.81
13.28
0.00
5.81
10.70
3.32
11.02
5.80
Time
(minutes)
29.31
16.50
5.41
9.17
>240
47.65
21.67
37.63
210.00
5.89
34.00
21.68
21.30
>240
5.80
149.00
15.55
42.09
0.57
15.00
6.82
34.59
14.67
17.17
11.48
10.13
>240
>240
41.03
10.25
1 pH of a 10% solution of the compound in water.
1120 - 12
Revision 0
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METHOD i.^O
DERMAL CORROSION
7.2.1 Add sample to
CDS.
Is
there a
color change in
the reaction
vessel.
Proceed wiTh
assay.
Is it
a liquid
or a
solid?
7.3.2.1 Add sample
to tube and read
color change.
7.3.2.2 Assign
catergories.
7.3.1.1 Add sample
to tube and read
color change.
7.3.2.3 Confirmation
Test.
7.3.1.2 Assign
categories.
7.3.1.3 Confirmation
test.
7.4.1.4 Heat assay
7.4.1.5 - 7.4.1.7
Apply solution
to membrane.
Use other method
to determine corrosivity.
1120 - 13
7.4.1.8 - 7.4.2.6 Add
one membrane and
apply to vial and
look for change.
7A.2.1 - 7.4.2.9 Add
3 more disks and
samples to vial and
check for color reactions
recording exact time
of color change.
7.4.2.10 Run positive
and negative controls.
7.4.3 Assignment of
Groups.
Revision 0
January 1995
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METHOD 9050A
SPECIFIC CONDUCTANCE
1.0 SCOPE AND APPLICATION
1.1 Method 9050 is used to measure the specific conductance of
drinking, ground, surface, and saline waters and domestic and industrial aqueous
wastes. Method 9050 is not applicable to solid samples or to organic samples.
2.0 SUMMARY OF METHOD
2.1 The specific conductance of a sample is measured using a self-
contained conductivity meter (Wheatstone bridge-type or equivalent).
2.2 Whenever possible, samples are analyzed at 25°C. If samples are
analyzed at different temperatures, temperature corrections must be made and
results reported at 25°C.
3.0 INTERFERENCES
3.1 Platinum electrodes can degrade and cause erratic results. When
this happens, as evidenced by erratic results or flaking off of the platinum
black, the electrode should be replatinized.
3.2 The specific conductance cell can become coated with oil and
other materials. It is essential that the cell be thoroughly rinsed and, if
necessary, cleaned between samples.
4.0 APPARATUS AND MATERIALS
4.1 Self-contained conductivity instruments: an instrument
consisting of a source of alternating current, a Wheatstone bridge, null
indicator, and a conductivity cell or other instrument measuring the ratio of
alternating current through the cell to voltage across it. The latter has the
advantage of a linear reading of conductivity. Choose an instrument capable of
measuring conductivity with an error not exceeding 1% or 1 uS/cm, whichever is
greater.
4.2 PI at i num-el ectrode or non-platinum-electrode specific conductance
cell.
4.3 Water bath.
4.4 Thermometer: capable of being read to the nearest 0.1°C and
covering the range 23"C to 27°C. An electrical thermometer having a small
thermistor sensing element is convenient because of its rapid response.
9050A - 1 Revision 1
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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 reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Conductivity water: Pass distilled water through a mixed-bed
deionizer and discard first 1,000 ml. Conductivity should be less than 1 uS/cm.
5.3 Standard potassium chloride (0.0100 M): Dissolve 0.7456 g
anhydrous KC1 in conductivity water and make up to 1,000 ml at 25°C. This
solution will have a specific conductance of 1,413 uS/cm at 25°C.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 All sample containers must be prewashed and thoroughly rinsed.
Both plastic and glass containers are suitable.
6.3 Aqueous samples should be stored at 4°C and analyzed within 28
days.
7.0 PROCEDURE
7.1 Determination of cell constant: Rinse conductivity cell with at
least three portions of 0.01 N KC1 solution. Adjust temperature of a fourth
portion to 25.0 + 0.1°C. Measure resistance of this portion and note
temperature. Compute cell constant, C:
C = (0.001413)(RKC1) 1 + 0.0191 (t - 25)
where:
= measured resistance, ohms; and
t = observed temperature, °C.
7.2 Conductivity measurement: Rinse cell with one or more portions
of sample. Adjust temperature of a final portion to 25.0 + 0.1°C. Measure
sample resistance or conductivity and note temperature.
9050A - 2 Revision 1
January 1995
4
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7.3 Calculation: The temperature coefficient of most waters is only
approximately the same as that of standard KC1 solution; the more the temperature
of measurement deviates from 25.0°C, the greater the uncertainty in applying the
temperature correction. Report all conductivities at 25.0°C.
7.3.1 When sample resistance is measured, conductivity at 25"C
i s:
K =
(1,000,000)(C)
Rm1 + 0.0191 (t - 25)
where:
K = conductivity, uS/cm;
C = cell constant, cm-L;
Rm = measured resistance of sample, ohms; and
t = temperature of measurement.
7.3.2 When sample conductivity is measured, conductivity at
25°C is:
(K )(1,000,000)(C)
K = m
I + 0.0191 CT^ Zb)
where:
Km = measured conductivity, uS at t°C, and other
units are defined as above.
NOTE: If conductivity readout is in uS/cm, delete the factor 1,000,000
in the numerator.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Analyze an independently prepared check standard to verify
calibration.
8.3 Analyze one duplicate sample for every 10 samples.
9050A - 3 Revision 1
January 1995
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9.0 METHOD PERFORMANCE
9.1 Three synthetic samples were tested with the following results:
Conduc-
tivity
uS/cm
147.0
303.0
228.0
No. of
Results
117
120
120
Relative
Standard
Deviation
%
8.6
7.8
8.4
Relative
Error
%
9.4
1.9
3.0
10.0 REFERENCES
1. Standard Methods for the Examination of Water and Wastewater, 16th ed.
(1985), Method 205.
9050A - 4 Revision 1
January 1995
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METHOD 9050A
SPECIFIC CONDUCTANCE
7.1 Determination of
cell constant.
>
r
7.2 Measure
conductivity;
note temperature.
>
r
7.3 Perform
calculations.
3
r
9050A - 5
Revision 1
January 1995
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METHOD 9095A
PAINT FILTER LIQUIDS TEST
1.0 SCOPE AND APPLICATION
1.1 This method is used to determine the presence of free liquids in a
representative sample of waste.
1.2 The method is used to determine compliance with 40 CFR 264.314 and
265.314.
2.0 SUMMARY OF METHOD
2.1 A predetermined amount of material is placed in a paint filter. If
any portion of the material passes through and drops from the filter within the
5-min test period, the material is deemed to contain free liquids.
3.0 INTERFERENCES
3.1 Filter media were observed to separate from the filter cone on
exposure to alkaline materials. This development causes no problem if the sample
is not disturbed.
3.2 Temperature can affect the test results if the test is performed
below the freezing point of any liquid in the sample. Tests must be performed
above the freezing point and can, but is not required to, exceed room temperature
of 25° C.
4.0 APPARATUS AND MATERIALS
4.1 Conical paint filter: Mesh number 60 +/- 5% (fine meshed size).
Available at local paint stores such as Sherwin-Williams and Glidden.
4.2 Glass funnel: If the paint filter, with the waste, cannot sustain
its weight on the ring stand, then a fluted glass funnel or glass funnel with a
mouth large enough to allow at least 1 in. of the filter mesh to protrude should
be used to support the filter. The funnel is to be fluted or have a large open
mouth in order to support the paint filter yet not interfere with the movement,
to the graduated cylinder, of the liquid that passes through the filter mesh.
4.3 Ring stand and ring, or tripod.
4.4 Graduated cylinder or beaker: 100-mL.
5.0 REAGENTS
5.1 None.
9095A - 1 Revision 1
January 1995
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6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected according to the directions in Chapter
Nine of this manual.
6.2 A 100-mL or 100-g representative sample is required for the test.
If it is not possible to obtain a sample of 100 mL or 100 g that is sufficiently
representative of the waste, the analyst may use larger size samples in multiples
of 100 mL or 100 g, i.e., 200, 300, 400 mL or g. However, when larger samples
are used, analysts shall divide the sample into 100-mL or 100-g portions and test
each portion separately. If any portion contains free liquids, the entire sample
is considered to have free liquids. If the sample is measured volumetrically,
then it should lack major air spaces or voids.
7.0 PROCEDURE
7.1 Assemble test apparatus as shown in Figure 1.
7.2 Place sample in the filter. A funnel may be used to provide support
for the paint filter. If the sample is of such light bulk density that it
overflow the filter, then the sides of the filter can be extended upward by
taping filter paper to the inside of the filter and above the mesh. Settling the
sample into the paint filter may be facilitated by lightly tapping the side of
the filter as it is being filled.
7.3 In order to assure uniformity and standardization of the test,
material such as sorbent pads or pillows which do not conform to the shape of the
paint filter, should be cut into small pieces and poured into the filter. Sample
size reduction may be accomplished by cutting the sorbent material with scissors,
shears, knife, or other such device so as to preserve as much of the original
integrity of the sorbent fabric as possible. Sorbents enclosed in a fabric
should be mixed with the resultant fabric pieces. The particles to be tested
should be reduced smaller than 1 cm (i.e., should be capable of passing through
a 9.5 mm (0.375 inch) standard sieve). Grinding sorbent materials should be
avoided as this may destroy the integrity of the sorbent and produce many "fine
particles" which would normally not be present.
7.4 For brittle materials larger than 1 cm that do not conform to the
filter, light crushing to reduce oversize particles is acceptable if it is not
practical to cut the material. Materials such as clay, silica gel, and some
polymers may fall into this category.
7.5 Allow sample to drain for 5 min into the graduated cylinder.
7.6 If any portion of the test material collects in the graduated
cylinder in the 5-min period, then the material is deemed to contain free liquids
for purposes of 40 CFR 264.314 and 265.314.
8.0 QUALITY CONTROL
8.1 Duplicate samples should be analyzed on a routine basis.
9095A - 2 Revision 1
January 1995
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9.0 METHOD PERFORMANCE
9.1 No data provided.
10.0 REFERENCES
10.1 None provided.
9095A - 3 Revision 1
January 1995
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RING STAND —
^-FUNNEL
/PAINT FILTER
i— GRADUATED CYLINDER
Figure 1. Paint filter test apparatus.
9095A - 4
Revision 1
January 1995
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METHOD 9095A
PAINT FILTER LIQUIDS TEST
Start
1
r
7.1 Assemble
test apparatus.
^
r
7.2 Place sample
in filter.
i
r
7.3 Allow sample
to drain into
graduated cylinder.
7.4 Did
any test
material collect
in graduated
cylinder?
7.4 Material is
deemed to contain
free liquids; see 40
CFR 264.314 or
265.314.
i
r
Stop
9095A - 5
Revision 1
January 1995
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CHAPTER EIGHT
METHODS FOR DETERMINING CHARACTERISTICS
Methods for determining the characterisitics of Ignitability for liquids,
Corrosivity for liquids, and Toxicity are included. Guidance for determining
Toxic Gas Generation is found in Chapter Seven, Sections 7.3.3 and 7.3.4.
EIGHT - 1 Revision 2
January 1995
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8.1 Ignitability
The following methods are found in Section 8.1:
Method 1010: Pensky-Martens Closed-Cup Method for Determining
Ignitability
Method 1020A: Setaflash Closed-Cup Method for Determining
Ignitability
EIGHT - 2 Revision 2
January 1995
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8.2 Corrosivity
The following methods are found in Section 8.2:
Method 9040B: pH Electrometric Measurement
Method 1110: Corrosivity Toward Steel
EIGHT - 3 Revision 2
Januarv 1995
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8.3 Reactivity
Refer to guidance given in Chapter Seven, especially Section 7.3.3 and
7.3.4.
EIGHT - 4 Revision 2
January 1995
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8.4 Toxicity
The following methods are found in Section 8.4:
Method 1310A: Extraction Procedure (EP) Toxicity Test Method
and Structural Integrity Test
Method 1311: Toxicity Characteristic Leaching Procedure
EIGHT - 5 Revision 2
January 1995
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CHAPTER TEN
SAMPLING METHODS
The following methods are found in Chapter Ten:
Method 0010:
Appendix A:
Appendix B:
Method 0011:
Method 0020:
Method 0023A:
Method 0030:
Method 0031:
Method 0040:
Method 0050:
Method 0051:
Method 0100:
Modified Method 5 Sampling Train
Preparation of XAD-2 Sorbent Resin
Total Chromatographable Organic Material
Analysis
Sampling for Formaldehyde Emissions from
Stationary Sources
Source Assessment Sampling System (SASS)
Sampling Method for Polychlorinated Dibenzo-p-
Dioxins and Polychlorinated Dibenzofuran
Emissions from Stationary Sources
Volatile Organic Sampling Train
Sampling Method for Volatile Organic Compounds
(SMVOC)
Sampling of Principal Organic Hazardous
Constituents from Combustion Sources Using
Tedlar® Bags
Isokinetic HC1/C12 Emission Sampling Train
Midget Impinger HC1/C12 Emission Sampling Train
Sampling for Formaldehyde and Other Carbonyl
Compounds in Indoor Air
SIX - 1
Revision 2
January 1995
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METHOD 0011
SAMPLING FOR FORMALDEHYDE EMISSIONS FROM STATIONARY SOURCES
1.0 SCOPE AND APPLICATION
1.1 This method is applicable to the determination of Destruction and
Removal Efficiency (ORE) of formaldehyde, CAS Registry number 50-00-0, and
possibly other aldehydes and ketones from stationary sources as specified in the
regulations. The methodology has been applied specifically to formaldehyde.
However, many laboratories have extended the application to other aldehydes and
ketones.
1.2 Compounds derivatized with 2,4-dinitrophenylhydrazine can be detected
as low as 6.4 x 10"8 Ibs/cu ft (1.8 ppbv) in stack gas over a 1 hour sampling
period, sampling approximately 45 cu ft.
1.3 This method is restricted to use by, or under the close supervision
of, analysts experienced in sampling organic compounds in air. Each analyst must
demonstrate the ability to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 Gaseous and particulate pollutants are withdrawn isokinetically from
an emission source and are collected in aqueous acidic 2,4-dinitrophenyl-
hydrazine. Formaldehyde present in the emissions reacts with the 2,4-dinitro-
phenylhydrazine to form the formaldehyde dinitrophenylhydrazone derivative. The
dinitrophenylhydrazone derivative is extracted, solvent-exchanged, concentrated,
and then analyzed by high performance liquid chromatography (HPLC).
3.0 INTERFERENCES
3.1 A decomposition product of 2,4-dinitrophenylhydrazine, 2,4-dinitro-
aniline, can be an analytical interferant if concentrations are high. 2,4-
Dinitroaniline can coelute with the 2,4-dinitrophenylhydrazone of formaldehyde
under high performance liquid chromatography conditions used for the analysis.
High concentrations of highly oxygenated compounds, especially acetone, that have
the same retention time or nearly the same retention time as the
dinitrophenylhydrazone of formaldehyde, and that also absorb at 360 nm, will
interfere with the analysis.
3.2 Formaldehyde, acetone, and 2,4-dinitroaniline contamination of the
aqueous acidic 2,4-dinitrophenylhydrazine (DNPH) reagent is frequently
encountered. The reagent must be prepared within five days of use in the field
and must be stored in an uncontaminated environment both before and after
sampling, in order to minimize blank problems. Some concentration of acetone
contamination is unavoidable, because acetone is ubiquitous in laboratory and
field operations. However, the acetone contamination must be minimized.
0011 - 1 Revision 0
January 1995
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4.0 APPARATUS AND MATERIALS
4.1 This sampling train configuration is adapted from EPA Method 5
procedures. The sampling train consists of the following components: Probe
nozzle, pitot tube, differential pressure gauge, metering system, barometer, and
gas density determination equipment. A schematic of the sampling train is shown
in Figure 1.
4.1.1 Probe Nozzle - The probe nozzle shall be quartz or glass with
sharp, tapered (30° angle) leading edge. The taper shall be on the
outside to preserve a constant inner diameter. The nozzle shall be
buttonhook or elbow design. A range of nozzle sizes suitable for
isokinetic sampling should be available in increments of 0.16 cm (1/16
in.), e.g., 0.32 to 1.27 cm (1/8 to 1/2 in.), or larger if higher volume
sampling trains are used. Each nozzle shall be calibrated according to
the procedures outlined in Sec. 8.1.
4.1.2 Probe Liner - Borosilicate glass or quartz shall be used for
the probe liner. The tester should not allow the temperature in the probe
to exceed 120 ±14'C (248 ±25'F).
4.1.3 Pitot Tube - The pitot tube shall be Type S or any other
appropriate device. The Type S pitot tube shall be made of metal tubing
(e.g., stainless steel). It is recommended that the external tubing
diameter be between 0.48 and 0.95 cm. There shall be an equal distance
from the base of each leg to its face-opening plane; it is recommended
that this distance be between 1.05 and 1.50 times the external tubing
diameter. The face openings of the pitot tube shall, preferably, be
aligned but slight misalignments of the openings are permissible. The
Type S pitot tube assembly shall have a known coefficient, determined as
outlined in Sec. 4 of EPA Method 2. The pitot tube shall be attached to
the probe to allow constant monitoring of the stack gas velocity. The
impact (high pressure) opening plane of the pitot tube shall be even with
or above the nozzle entry plane (see EPA Method 2, Figure 2-6b) during
sampling.
4.1.4 Differential Pressure Gauge - The differential pressure gauge
shall be an inclined manometer or equivalent device as described in Sec.
2.2 of EPA Method 2. One manometer shall be used for velocity-head
readings and the other for orifice differential pressure readings.
4.1.5 Impingers - The sampling train requires a minimum of four
impingers, connected as shown in Figure 1, with ground glass (or
equivalent) vacuum-tight fittings. For the first, third, and fourth
impingers, use the Greenburg-Smith design, modified by replacing the tip
with a 1.3 cm inside diameter (1/2 in.) glass tube extending to 1.3 cm
(1/2 in.) from the bottom of the flask. For the second impinger, use a
Greenburg-Smith impinger with the standard tip. Place a thermometer
capable of measuring temperature to within 1°C (2°F) at the outlet of the
fourth impinger for monitoring purposes.
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4.1.6 Metering System - The necessary components of the metering
system are a vacuum gauge, leak-free pump, thermometers capable of
measuring temperature within 3°C (5.4°F), dry-gas meter capable of
measuring volume to within 1%, and related equipment as shown in Figure 1.
At a minimum, the pump should be capable of 4 cfm free flow, and the dry
gas meter should have a recording capacity of 0-999.9 cu ft with a
resolution of 0.005 cu ft. Other metering systems may be used which are
capable of maintaining sample volumes to within 2%. The metering system
may be used in conjunction with a pitot tube to enable checks of
isokinetic sampling rates.
4.1.7 Barometer - The barometer may be mercury, aneroid, or other
barometer capable of measuring atmospheric pressure to within 2.5 mm Hg
(0.1 in. Hg). In many cases, the barometric reading may be obtained from
a nearby National Weather Service Station, in which case the station value
(which is the absolute barometric pressure) is requested and an adjustment
for elevation differences between the weather station and sampling point
is applied at a rate of minus 2.5 mm Hg (0.1 in. Hg) per 30 m (100 ft)
elevation increase (vice versa for elevation decrease).
4.1.8 Gas Density Determination Equipment - The gas density
determination equipment includes a temperature sensor and pressure gauge
(as described in Sections 2.3 and 2.4 of EPA Method 2), and gas analyzer,
if necessary (an Orsat of Fyrite type combustion gas analyzer, or
equivalent. For analyzer maintenance and operation procedures, follow the
instructions recommended by the manufacturer). The temperature sensor
ideally should be permanently attached to the pitot tube or sampling probe
in a fixed configuration such that the tip of the sensor extends beyond
the leading edge of the probe sheath and does not touch any metal.
Alternatively, the sensor may be attached just prior to use in the field.
Note, however, that if the temperature sensor is attached in the field,
the sensor must be placed in an interference-free arrangement with respect
to the Type S pitot tube openings (see EPA Method 2, Figure 2-7). As a
second alternative, if a difference of no more than 1% in the average
velocity measurement is to be introduced, the temperature gauge need not
be attached to the probe or pitot tube.
4.2 Sample Recovery
4.2.1 Probe Liner - Probe nozzle and brushes; Teflon® bristle
brushes with stainless steel wire handles are required. The probe brush
shall have extensions of stainless steel, Teflon®, or inert material at
least as long as the probe. The brushes shall be properly sized and
shaped to brush out the probe liner, the probe nozzle, and the impingers.
4.2.2 Wash Bottles - Three wash bottles are required. Teflon® or
glass wash bottles are recommended. Polyethylene wash bottles should not
be used because organic contaminants may be extracted by exposure to the
organic solvents used for sample recovery.
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4.2.3 Graduated Cylinder and/or Balance - A graduated cylinder or
balance is required to measure condensed water to the nearest 1 mL or 1 g.
Graduated cylinders shall have divisions not greater than 2 mL.
Laboratory balances capable of weighing to ±0.5 g are required.
4.2.4 Amber Glass Storage Containers - One-liter wide-mouth amber
flint glass bottles with Teflon®-!ined caps are required to store impinger
water samples. The bottles must be sealed with Teflon® tape.
4.2.5 Rubber Policeman and Funnel - A rubber policeman and funnel
are required to aid in the transfer of materials into and out of
containers in the field.
4.3 Reagent Preparation
4.3.1 Bottles/Caps - Amber 1 - 4 L bottles with Teflon®-!ined caps
are required for storing cleaned DNPH solution. Additional 4-L bottles
are required to collect waste organic solvents.
4.3.2 Large Glass Container - At least one large glass (8 to 16 L)
is required for mixing the aqueous acidic DNPH solution.
4.3.3 Stir Plate/Large Stir Bars/Stir Bar Retriever - A magnetic
stir plate and large stir bar are required for the mixing of the aqueous
acidic DNPH solution. A stir bar retriever is needed for removing the
stir bar from the large container holding the DNPH solution.
4.3.4 Buchner Filter/Filter Flask/Filter Paper - A large filter
flask (2-4 L) with a buchner filter, appropriate rubber stopper, filter
paper, and connecting tubing are required for filtering the aqueous acidic
DNPH solution prior to cleaning.
4.3.5 Separatory Funnel - At least one large separatory funnel
(2 L) is required for cleaning the DNPH prior to use.
4.3.6 Beakers - Beakers (150 mL, 250 mL, and 400 mL) are useful for
holding/measuring organic liquids when cleaning the aqueous acidic DNPH
solution and for weighing DNPH crystals.
4.3.7 Funnels - At least one large funnel is needed for pouring the
aqueous acidic DNPH into the separatory funnel.
4.3.8 Graduated Cylinders - At least one large graduated cylinder
(1 to 2 L) is required for measuring organic-free reagent water and acid
when preparing the DNPH solution.
4.3.9 Top-loading Balance - A one-place top loading balance is
needed for weighing out the DNPH crystals used to prepare the aqueous
acidic DNPH solution.
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4.3.10 Spatulas - Spatulas are needed for weighing out DNPH when
preparing the aqueous DNPH solution.
4.4 Crushed Ice - Quantities of crushed ice ranging from 10-50 Ib may be
necessary during a sampling run, depending upon ambient temperature. Samples
which have been taken must be stored and shipped cold; sufficient ice for this
purpose must be allowed.
5.0 REAGENTS
5.1 Reagent Grade Chemicals - 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 Silica Gel - Silica gel shall be indicating type, 6-16 mesh. If the
silica gel has been used previously, dry at 175°C (350°F) for 2 hours before
using. New silica gel may be used as received. Alternatively, other types of
desiccants (equivalent or better) may be used.
5.4 2,4-Dinitrophenylhydrazine (DNPH), [2,4-(02N)2C6H3]NHNH2 - The quantity
of water may vary from 10 to 30%.
5.4.1 The 2,4-dinitrophenylhydrazine reagent must be prepared in
the laboratory within five days of sampling use in the field. Preparation
of DNPH can also be done in the field, with consideration of appropriate
procedures required for safe handling of solvent in the field. When a
container of prepared DNPH reagent is opened in the field, the contents of
the opened container should be used within 48 hours. All laboratory
glassware must be washed with detergent and water and rinsed with water,
methanol, and methylene chloride prior to use.
NOTE: DNPH crystals and DNPH solution are potential carcinogens and should be
handled with plastic gloves at all times, with prompt and extensive use of
running water in case of skin exposure.
5.4.2 Preparation of Aqueous Acidic DNPH Derivatizing Reagent -
Each batch of DNPH reagent should be prepared and purified within five
days of sampling, according to the procedure described below.
NOTE: Reagent bottles for storage of cleaned DNPH derivatizing solution must be
rinsed with acetonitrile and dried before use. Baked glassware is not
essential for preparation of DNPH reagent. The glassware must not be
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rinsed with acetone or an unacceptable concentration of acetone
contamination will be introduced. If field preparation of DNPH is
performed, caution must be exercised in avoiding acetone contamination.
5.4.2.1 Place an 8 L container under a fume hood on a
magnetic stirrer. Add a large stir bar and fill the container half
full of organic-free reagent water. Save the empty bottle from the
organic-free reagent water. Start the stirring bar and adjust the
stir rate to be as fast as possible. Using a graduated cylinder,
measure 1.4 L of concentrated hydrochloric acid. Slowly pour the
acid into the stirring water. Fumes may be generated and the water
may become warm. Weigh the DNPH crystals to ±0.1 g (see Table 1 for
approximate amounts) and add to the stirring acid solution. Fill
the 8 L container to the 8 L mark with organic-free reagent water
and stir overnight. If all of the DNPH crystals have dissolved
overnight, add additional DNPH and stir for two more hours.
Continue the process of adding DNPH with additional stirring until
a saturated solution has been formed. Filter the DNPH solution
using vacuum filtration. Gravity filtration may be used, but a much
longer time is required. Store the filtered solution in an amber
bottle at room temperature.
5.4.2.2 Within five days of proposed use, place about 1.6 L
of the DNPH reagent in a 2 L separatory funnel. Add approximately
200 ml of methylene chloride and stopper the funnel. Wrap the
stopper of the funnel with paper towels to absorb any leakage.
Invert and vent the funnel. Then shake vigorously for 3 minutes.
Initially, the funnel should be vented frequently (every 10 - 15
seconds). After the layers have separated, discard the lower
(organic) layer.
5.4.2.3 Extract the DNPH a second time with methylene
chloride and finally with cyclohexane. When the cyclohexane layer
has separated from the DNPH reagent, the cyclohexane layer will be
the top layer in the separatory funnel. Drain the lower layer (the
cleaned extracted DNPH reagent solution) into an amber bottle that
has been rinsed with acetonitrile and allowed to dry.
5.4.3 DNPH Reagent Check - Take two aliquots of the extracted DNPH
reagent. The size of the aliquots is dependent upon the exact sampling
procedure used, but 100 ml is reasonably representative. Analyze one
aliquot of the reagent according to the procedure of Method 8315 as a
Quality Control check to ensure that the background in the reagent is
acceptable for field use. Save the other aliquot of aqueous acidic DNPH
for use as a method blank when the analysis is performed.
5.4.4 Shipment to the Field - Tightly cap the bottle containing
extracted DNPH reagent using a Teflon®-!ined cap. Seal the bottle with
Teflon® tape. After the bottle is labeled, the bottle may be placed in a
friction-top can (paint can or equivalent) containing a 1-2 inch layer of
granulated charcoal and stored at ambient temperature until use.
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5.4.4.1 If the DNPH reagent has passed the Quality Control
criteria, the reagent may be packaged to meet necessary shipping
requirements and sent to the sampling area. If the Quality Control
criteria are not met, the reagent solution may be re-extracted or
the solution may be re-prepared and the extraction sequence
repeated.
5.4.4.2 If the DNPH reagent is not used in the field within
five days of extraction, an aliquot may be taken and analyzed as
described in Method 8315. If the reagent meets the Quality Control
requirements, the reagent may be used. If the reagent does not meet
the Quality Control requirements, the reagent must be discarded and
new reagent must be prepared and tested.
5.4.5 Calculation of Acceptable Concentrations of Impurities in
DNPH Reagent - The acceptable impurity concentration (AIC, ng/ml) is
calculated from the expected analyte concentration in the sampled gas
(EAC, ppbv), the volume of air that will be sampled at standard conditions
(SVOL, L), the formula weight of the analyte (FW, g/mol), and the volume
of DNPH reagent that will be used in the impingers (RVOL, ml):
AIC = 0.1 x [EAC x SVOL x FW/24.4 x (FW + 180J/FW](RVOL/1,000)
where:
0.1 is the acceptable contaminant concentration,
24.4 is a factor relating ppbv to g/L,
180 is a factor relating underivatized to derivatized analyte
1,000 is a unit conversion factor.
5.4.6 Disposal of Excess DNPH Reagent - Excess DNPH reagent may be
returned to the laboratory and recycled or treated as aqueous waste for
disposal purposes. 2,4-Dinitrophenylhydrazine is a flammable solid when
dry, so water should not be evaporated from the solution of the reagent.
5.5 Field Spike Standard Preparation - To prepare a formaldehyde field
spiking standard at 4010 mg/L, use a 500 juL syringe to transfer 0.5 ml of 37%
by weight of formaldehyde (401 g/L) to a 50 mL volumetric flask containing
approximately 40 mL of methanol. Dilute to 50 mL with methanol.
5.6 Hydrochloric Acid, HC1 - Reagent grade hydrochloric acid
(approximately 12N) is required for acidifying the aqueous DNPH solution.
5.7 Methylene Chloride, CH2C12 - Methylene chloride (suitable for residue
and pesticide analysis, GC/MS, HPLC, GC, Spectrophotometry or equivalent) is
required for cleaning the aqueous acidic DNPH solution, rinsing glassware, and
recovery of sample trains.
5.8 Cyclohexane, C6H12 - Cyclohexane (HPLC grade) is required for cleaning
the aqueous acidic DNPH solution.
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NOTE: Do not use spectroanalyzed grades of cyclohexane if this sampling
methodology is extended to aldehydes and ketones with four or more carbon
atoms.
5.9 Methanol, CH3OH - Methanol (HPLC grade or equivalent) is necessary for
rinsing glassware.
5.10 Acetonitrile, CH3CN - Acetonitrile (HPLC grade or equivalent) is
required for rinsing glassware.
5.11 Formaldehyde, HCHO - Formaldehyde (analytical reagent grade, or
equivalent) is required for preparation of standards. If other aldehydes or
ketones are used, analytical reagent grade, or equivalent, is required.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Because of the complexity of this method, field personnel should be
trained in and experienced with the test procedures in order to obtain reliable
results.
6.2 Laboratory Preparation
6.2.1 All the components shall be maintained and calibrated
according to the procedure described in APTD-0576 (Air Pollution Technical
Document, see references), unless otherwise specified.
6.2.2 Weigh several 200 to 300 g portions of silica gel in airtight
containers to the nearest 0.5 g. Record on each container the total
weight of the silica gel plus containers. As an alternative to
preweighing the silica gel, it may instead be weighed directly in the
impinger or sampling holder just prior to train assembly.
6.3 Preliminary Field Determinations
6.3.1 Select the sampling site and the minimum number of sampling
points according to EPA Method 1 or other relevant criteria. Determine
the stack pressure, temperature, and range of velocity heads using EPA
Method 2. A leak-check of the pitot lines according to EPA Method 2, Sec.
3.1, must be performed. Determine the stack gas moisture content using
EPA Approximation Method 4 or its alternatives to establish estimates of
isokinetic sampling-rate settings. Determine the stack gas dry molecular
weight, as described in EPA Method 2, Sec. 3.6. If integrated EPA Method
3 sampling is used for molecular weight determination, the integrated bag
sample shall be taken simultaneously with, and for the same total length
of time as, the sample run.
6.3.2 Select a nozzle size based on the range of velocity heads so
that it is not necessary to change the nozzle size in order to maintain
isokinetic sampling rates below 28 L/min (1.0 cfm). During the run, do
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not change the nozzle. Ensure that the proper differential pressure gauge
is chosen for the range of velocity heads encountered (see Sec. 2.2 of EPA
Method 2).
6.3.3 Select a suitable probe liner and probe length so that all
traverse points can be sampled. For large stacks, to reduce the length of
the probe, consider sampling from opposite sides of the stack.
6.3.4 A minimum of 45 ft3 of sample volume is required for the
determination of the Destruction and Removal Efficiency (ORE) of
formaldehyde from incineration systems (45 ft3 is equivalent to one hour
of sampling at 0.75 dscf). Additional sample volume shall be collected as
necessitated by the capacity of the DNPH reagent and analytical detection
limit constraints. To determine the minimum sample volume required, refer
to sample calculations in Sec. 10.0.
6.3.5 Determine the total length of sampling time needed to obtain
the identified minimum volume by comparing the anticipated average
sampling rate with the volume requirement. Allocate the same time to all
traverse points defined by EPA Method 1. To avoid timekeeping errors, the
length of time sampled at each traverse point should be an integer or an
integer plus 0.5 min.
6.3.6 In some circumstances (e.g., batch cycles) it may be
necessary to sample for shorter times at the traverse points and to obtain
smaller gas-volume samples. In these cases, careful documentation must be
maintained in order to allow accurate calculation of concentrations.
6.4 Preparation of Collection Train
6.4.1 During preparation and assembly of the sampling train, keep
all openings where contamination can occur covered with Teflon® film or
aluminum foil until just prior to assembly or until sampling is about to
begin.
NOTE: Appendix A at the end of this procedure contains guidance on the addition
of a filter as a check on the survival of particulate material through the
impinger system. This filter can be added to the impinger train either
after the second impinger or after the third impinger.
6.4.2 Place 100 ml of cleaned DNPH solution in each of the first
two impingers, and leave the third impinger empty. If additional capacity
is required for high expected concentrations of formaldehyde in the stack
gas, 200 ml of DNPH per impinger may be used or additional impingers may
be used for sampling. Transfer approximately 200 to 300 g of pre-weighed
silica gel from its container to the fourth impinger. Care should be
taken to ensure that the silica gel is not entrained and carried out from
the impinger during sampling. Place the silica gel container in a clean
place for later use in the sample recovery. Alternatively, the weight of
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the silica gel plus impinger may be determined to the nearest 0.5 g and
recorded.
6.4.3 With a glass or quartz liner, install the selected nozzle
using a Viton-A 0-ring when stack temperatures are less than 260°C (500°F)
and a woven glass-fiber gasket when temperatures are higher. See APTD-
0576 (Rom, 1972) for details. Other connecting systems utilizing either
316 stainless steel or Teflon® ferrules may be used. Mark the probe with
heat-resistant tape or by some other method to denote the proper distance
into the stack or duct for each sampling point.
6.4.4 Assemble the train as shown in Figure 1. During assembly, do
not use any silicone grease on ground-glass joints upstream of the
impingers. Use Teflon® tape, if required. A very light coating of
silicone grease may be used on ground-glass joints downstream of the
impingers, but the silicone grease should be limited to the outer portion
(see APTD-0576) of the ground-glass joints to minimize silicone grease
contamination. If necessary, Teflon® tape may be used to seal leaks.
Connect all temperature sensors to an appropriate potentiometer/ display
unit. Check all temperature sensors at ambient temperature.
6.4.5 Place crushed ice all around the impingers.
6.4.6 Turn on and set the probe heating system at the desired
operating temperature. Allow time for the temperature to stabilize.
6.5 Leak-Check Procedures
6.5.1 Pre-test Leak Check
6.5.1.1 After the sampling train has been assembled, turn on
and set the probe heating system at the desired operating
temperature. Allow time for the temperature to stabilize. If a
Viton-A 0-ring or other leak-free connection is used in assembling
the probe nozzle to the probe liner, leak-check the train at the
sampling site by plugging the nozzle and pulling a 381 mm Hg (15 in.
Hg) vacuum.
NOTE: A lower vacuum may be used, provided that the lower vacuum is not exceeded
during the test.
6.5.1.2 If an asbestos string is used, do not connect the
probe to the train during the leak check. Instead, leak-check the
train by first attaching a carbon-filled leak check impinger to the
inlet and then plugging the inlet and pulling a 381 mm Hg (15 in.
Hg) vacuum. (A lower vacuum may be used if this lower vacuum is not
exceeded during the test.) Then connect the probe to the train and
leak-check at about 25 mm Hg (1 in. Hg) vacuum. Alternatively,
leak-check the probe with the rest of the sampling train in one step
at 381 mm Hg (15 in. Hg) vacuum. Leakage rates no greater than 4%
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of the average sampling rate or less than or equal to 0.00057 m3/roiri
(0.02 cfm), whichever is less, are acceptable.
6.5.1.3 The following leak check instructions for the
sampling train described in APTD-0576 and APTD-0581 may be helpful.
Start the pump with the fine-adjust valve fully open and coarse-
adjust valve completely closed. Partially open the coarse-adjust
valve and slowly close the fine-adjust valve until the desired
vacuum is reached. Do not reverse direction of the fine-adjust
valve, as liquid will back up into the train. If the desired vacuum
is exceeded, either perform the leak check at this higher vacuum or
end the leak check, as shown below, and start over.
6.5.1.4 When the leak check is completed, first slowly remove
the plug from the inlet to the probe. When the vacuum drops to 127
mm (5 in.) Hg or less, immediately close the coarse-adjust valve.
Switch off the pumping system and reopen the fine-adjust valve. Do
not reopen the fine-adjust valve until the coarse-adjust valve has
been closed to prevent the liquid in the impingers from being forced
backward into the sampling line and silica gel from being entrained
backward into the third impinger.
6.5.2 Sampling Run Leak Check
6.5.2.1 If, during the sampling run, a component change
(i.e., impinger) becomes necessary, a leak check shall be conducted
immediately after the interruption of sampling and before the change
is made. The leak check shall be done according to the procedure
described in Sec. 6.5.1, except that it shall be done at a vacuum
greater than or equal to the maximum value recorded up to that point
in the test. If the leakage rate is found to be no greater than
0.00057 m3/min (0.02 cfm) or 4% of the average sampling rate
(whichever is less), the results are acceptable. If a higher
leakage rate is obtained, the tester must void the sampling run.
NOTE: Any correction of the sample volume by calculation reduces the integrity
of the pollutant concentration data generated and must be avoided.
6.5.2.2 Immediately after a component change and before
sampling is reinitiated, a leak check similar to a pre-test leak
check must also be conducted.
6.5.3 Post-test Leak Check - A leak check is mandatory at the
conclusion of each sampling run. The leak check shall be done with the
same procedures as the pre-test leak check, except that the post-test leak
check shall be conducted at a vacuum greater than or equal to the maximum
value reached during the sampling run. If the leakage rate is found to be
no greater than 0.00057 m3/min (0.02 cfm) or 4% of the average sampling
rate (whichever is less), the results are acceptable. If, however, a
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higher leakage rate is obtained, the tester shall record the leakage rate
and void the sampling run.
6.6 Sampling Train Operation
6.6.1 During the sampling run, maintain an isokinetic sampling rate
to within 10% of true isokinetic, below 28 L/min (1.0 cfm). Maintain a
temperature around the probe of 120°C (248° ±25°F).
6.6.2 For each run, record the data on a data sheet such as the one
shown in Figure 2. Be sure to record the initial dry-gas meter reading.
Record the dry-gas meter readings at the beginning and end of each
sampling time increment, when changes in flow rates are made, before and
after each leak check, and when sampling is halted. Take other readings
required by Figure 2 at least once at each sample point during each time
increment and additional readings when significant adjustments (20%
variation in velocity head readings) necessitate additional adjustments in
flow rate. Level and zero the manometer. Because the manometer level and
zero may drift due to vibrations and temperature changes, make periodic
checks during the traverse.
6.6.3 Clean the stack access ports prior to the test run to
eliminate the chance of sampling deposited material. To begin sampling,
remove the nozzle cap, verify that the filter and probe heating systems
are at the specified temperature, and verify that the pitot tube and probe
are properly positioned. Position the nozzle at the first traverse point,
with the tip pointing directly into the gas stream. Immediately start the
pump and adjust the flow to isokinetic conditions. Nomographs, which aid
in the rapid adjustment of the isokinetic sampling rate without excessive
computations, are available. These nomographs are designed for use when
the Type S pitot tube coefficient is 0.84 ±0.02 and the stack gas
equivalent density (dry molecular weight) is equal to 29 ±4. APTD-0576
details the procedure for using the nomographs. If the stack gas
molecular weight and the pitot tube coefficient are outside the above
ranges, do not use the nomographs unless appropriate steps are taken to
compensate for the deviations.
6.6.4 When the stack is under significant negative pressure
(equivalent to the height of the impinger stem), take care to close the
coarse-adjust valve before inserting the probe into the stack in order to
prevent liquid from backing up through the train. If necessary, the pump
may be turned on with the coarse-adjust valve closed.
6.6.5 When the probe is in position, block off the openings around
the probe and stack access port to prevent nonrepresentative dilution of
the gas stream.
6.6.6 Traverse the stack cross section, as required by EPA Method
1, being careful not to bump the probe nozzle into the stack walls when
sampling near the walls or when removing or inserting the probe through
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the access port, in order to minimize the chance of extracting deposited
material.
6.6.7 During the test run, make periodic adjustments to keep the
temperature around the probe at the proper levels. Add more ice and, if
necessary, salt, to maintain a temperature of less than 20°C (68°F) at the
silica gel outlet. Also, periodically check the level and zero of the
manometer.
6.6.8 A single train shall be used for the entire sampling run,
except in cases where simultaneous sampling is required in two or more
separate ducts or at two or more different locations within the same duct,
or in cases where equipment failure necessitates a change of trains. An
additional train or additional trains may also be used for sampling when
the capacity of a single train is exceeded.
6.6.9 When two or more trains are used, separate analyses of
components from each train shall be performed. If multiple trains have
been used because the capacity of a single train would be exceeded, first
impingers from each train may be combined, and second impingers from each
train may be combined.
6.6.10 At the end of the sampling run, turn off the coarse-adjust
valve, remove the probe and nozzle from the stack, turn off the pump,
record the final dry gas meter reading, and conduct a post-test leak
check. Also, leak check the pitot lines as described in EPA Method 2.
The lines must pass this leak check in order to validate the velocity-head
data.
6.6.11 Calculate percent isokineticity (see Method 2) to determine
whether the run was valid or another test should be made.
7.0 SAMPLE RECOVERY AND PREPARATION FOR ANALYSIS
7.1 Preparation
7.1.1 Proper cleanup procedure begins as soon as the probe is
removed from the stack at the end of the sampling period. Allow the probe
to cool. When the probe can be handled safely, wipe off all external
particulate matter near the tip of the probe nozzle and place a cap over
the tip to prevent losing or gaining particulate matter. Do not cap the
probe tip tightly while the sampling train is cooling because a vacuum
will be created, drawing liquid from the impingers back through the
sampling train.
7.1.2 Before moving the sampling train to the cleanup site, remove
the probe from the sampling train and cap the open outlet, being careful
not to lose any condensate that might be present. Remove the umbilical
cord from the last impinger and cap the impinger. If a flexible line is
used, let any condensed water or liquid drain into the impingers. Cap off
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any open impinger inlets and outlets. Ground glass stoppers, Teflon®
caps, or caps of other inert materials may be used to seal all openings.
7.1.3 Transfer the probe and impinger assembly to an area that is
clean and protected from wind so that the chances of contaminating or
losing the sample are minimized.
7.1.4 Inspect the train before and during disassembly, and note any
abnormal conditions.
7.1.5 Save a portion of all washing solution (methylene chloride,
water) used for cleanup as a blank. Transfer 200 ml of each solution
directly from the wash bottle being used and place each in a separate,
pre-labeled sample container.
7.2 Sample Containers
7.2.1 Container 1 - Probe and Impinger Catches. Using a graduated
cylinder, measure to the nearest ml, and record the volume of the solution
in the first three impingers. Alternatively, the solution may be weighed
to the nearest 0.5 g. Include any condensate in the probe in this
determination. Transfer the impinger solution from the graduated cylinder
into the amber flint glass bottle. Taking care that dust on the outside
of the probe or other exterior surfaces does not get into the sample,
clean all surfaces to which the sample is exposed (including the probe
nozzle, probe fitting, probe liner, first impinger, and impinger
connector) with methylene chloride. Use less than 500 ml for the entire
wash (250 ml would be better, if possible). Add the washings to the
sample container.
7.2.1.1 Carefully remove the probe nozzle and rinse the
inside surface with methylene chloride from a wash bottle. Brush
with a Teflon® bristle brush, and rinse until the rinse shows no
visible particles or yellow color, after which make a final rinse of
the inside surface. Brush and rinse the inside parts of the
Swagelok fitting with methylene chloride in a similar way.
7.2.1.2 Rinse the probe 1iner with methylene chloride. While
squirting the methylene chloride into the upper end of the probe,
tilt and rotate the probe so that all inside surfaces will be wetted
with methylene chloride. Let the methylene chloride drain from the
lower end into the sample container. The tester may use a funnel
(glass or polyethylene) to aid in transferring the liquid washes to
the container. Following the rinse with a Teflon® brush. Hold the
probe in an inclined position, and squirt methylene chloride into
the upper end as the probe brush is being pushed with a twisting
action through the probe. Hold the sample container underneath the
lower end of the probe, and catch any methylene chloride, water, and
particulate matter that is brushed from the probe. Run the brush
through the probe three times or more. With stainless steel or
other metal probes, run the brush through in the above prescribed
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manner at least six times since there may be small crevices in which
particulate matter can be entrapped. Rinse the brush with methylene
chloride or water, and quantitatively collect these washings in the
sample container. After the brushings, make a final rinse of the
probe as described above.
NOTE: Between sampling runs, brushes must be kept clean and free from
contamination.
7.2.1.3 Rinse the inside surface of each of the first three
impingers (and connecting tubing) three separate times. Use a small
portion of methylene chloride for each rinse, and brush each surface
to which sample is exposed with a Teflon® bristle brush to ensure
recovery of fine particulate matter. Water will be required for the
recovery of the impingers in addition to the specified quantity of
methylene chloride. There will be at least two phases in the
impingers. This two-phase mixture does not pour well, and a
significant amount of the impinger catch will be left on the walls.
The use of water as a rinse makes the recovery quantitative. Make
a final rinse of each surface and of the brush, using both methylene
chloride and water.
7.2.1.4 After all methylene chloride and water washings and
particulate matter have been collected in the sample container,
tighten the lid so that solvent, water, and DNPH reagent will not
leak out when the container is snipped to the laboratory. Mark the
height of the fluid level to determine whether leakage occurs during
transport. Seal the container with Teflon® tape. Label the
container clearly to identify its contents.
7.2.1.5 If the first two impingers are to be analyzed
separately to check for breakthrough, separate the contents and
rinses of the two impingers into individual containers. Care must
be taken to avoid physical carryover from the first impinger to the
second. The formaldehyde hydrazone is a solid which floats and
froths on top of the impinger solution. Any physical carryover of
collected moisture into the second impinger will invalidate a
breakthrough assessment.
7.2.2 Container 2 - Sample Blank. Prepare a sample blank by using
an amber flint glass container and adding a volume of DNPH reagent and
methylene chloride equal to the total volume in Container 1. Process the
blank in the same manner as Container 1.
7.2.3 Container 3 - Silica Gel. Note the color of the indicating
silica gel to determine whether it has been completely spent and make a
notation of its condition. The impinger containing the silica gel may be
used as a sample transport container with both ends sealed with tightly
fitting caps or plugs. Ground-glass stoppers or Teflon® caps may be used.
The silica gel impinger should then be labeled, covered with aluminum
foil, and packaged on ice for transport to the laboratory. If the silica
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gel is removed from the impinger, the tester may use a funnel to pour the
silica gel and a rubber policeman to remove the silica gel from the
impinger. It is not necessary to remove the small amount of dust
particles that may adhere to the impinger wall and are difficult to
remove. Since the gain in weight is to be used for moisture calculations,
do not use water or other liquids to transfer the silica gel. If a
balance is available in the field, the spent silica gel (or silica gel
plus impinger) may be weighed to the nearest 0.5 g.
7.2.4 Sample containers should be placed in a cooler, cooled by
although not in contact with ice. Sample containers must be placed
vertically and, since they are glass, protected from breakage during
shipment. Samples should be cooled during shipment so they will be
received cold at the laboratory.
8.0 CALIBRATION
8.1 Probe Nozzle - Probe nozzles shall be calibrated before their initial
use in the field. Using a micrometer, measure the inside diameter of the nozzle
to the nearest 0.025 mm (0.001 in.). Make measurements at three separate places
across the diameter and obtain the average of the measurements. The difference
between the high and low numbers shall not exceed 0.1 mm (0.004 in.). When the
nozzles become nicked or corroded, they shall be replaced and calibrated before
use. Each nozzle must be permanently and uniquely identified.
8.2 Pitot Tube - The Type S pitot tube assembly shall be calibrated
according to the procedure outlined in Sec. 4 of EPA Method 2, or assigned a
nominal coefficient of 0.84 if it is not visibly nicked or corroded and if it
meets design and intercomponent spacing specifications.
8.3 Metering System
8.3.1 Before its initial use in the field, the metering system
shall be calibrated according to the procedure outlined in APTD-0576.
Instead of physically adjusting the dry-gas meter dial readings to
correspond to the wet-test meter readings, calibration factors may be used
to correct the gas meter dial readings mathematically to the proper
values. Before calibrating the metering system, it is suggested that a
leak check be conducted. For metering system having diaphragm pumps, the
normal leak check procedure will not detect leakages within the pump. For
these cases, the following leak check procedure will apply: make a ten-
minute calibration run at 0.00057 m3/min (0.02 cfm). At the end of the
run, take the difference of the measured wet-test and dry-gas meter
volumes and divide the difference by 10 to get the leak rate. The leak
rate should not exceed 0.00057 m3/min (0.02 cfm).
8.3.2 After each field use, check the calibration of the metering
system by performing three calibration runs at a single intermediate
orifice setting (based on the previous field test). Set the vacuum at the
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maximum value reached during the test series. To adjust the vacuum,
insert a valve between the wet-test meter and the inlet of the metering
system. Calculate the average value of the calibration factor. If the
calibration has changed by more than 5%, recalibrate the meter over the
full range of orifice settings, as outlined in APTD-0576.
8.3.3 Leak Check of Metering System - The portion of the sampling
train from the pump to the orifice meter (see Figure 1) should be leak-
checked prior to initial use and after each shipment. Leakage after the
pump will result in less volume being recorded than is actually sampled.
Use the following procedure: Close the main valve on the meter box.
Insert a one-hole rubber stopper with rubber tubing attached into the
orifice exhaust pipe. Disconnect and vent the low side of the orifice
manometer. Close off the low side orifice tap. Pressurize the system to
13 - 18 cm (5 - 7 in.) water column by blowing into the rubber tubing.
Pinch off the tubing and observe the manometer for 1 min. A loss of
pressure on the manometer indicates a leak in the meter box. Leaks must
be corrected.
NOTE: If the dry-gas-meter coefficient values obtained before and after a test
series differ by greater than 5%, either the test series must be voided or
calculations for test series must be performed using whichever meter
coefficient value (i.e., before or after) gives the lower value of total
sample volume.
8.4 Probe Heater - The probe heating system must be calibrated before its
initial use in the field according to the procedure outlined in APTD-0576.
Probes constructed according to APTD-0581 need not be calibrated if the
calibration curves in APTD-0576 are used.
8.5 Temperature Gauges - Each thermocouple must be permanently and
uniquely marked on the casting. All mercury-in-glass reference thermometers must
conform to ASTM E-l 63C or 63F (American Society for Testing and Materials)
specifications. Thermocouples should be calibrated in the laboratory with and
without the use of extension leads. If extension leads are used in the field,
the thermocouple readings at the ambient air temperatures, with and without the
extension lead, must be noted and recorded. Correction is necessary if the use
of an extension lead produces a change greater than 1.5%.
8.5.1 Impinger and Dry-gas Meter Thermocouples - For the
thermocouples used to measure the temperature of the gas leaving the
impinger train, a three-point calibration at ice water, room air, and
boiling water temperatures is necessary. Accept the thermocouples only if
the readings at all three temperatures agree to ±2°C (3.6°F) with those of
the absolute value of the reference thermometer.
8.5.2 Probe and Stack Thermocouple - For the thermocouples used to
indicate the probe and stack temperatures, a three-point calibration at
ice water, boiling water, and hot oil bath temperatures must be performed.
Use of a point at room air temperature is recommended. The thermometer
and thermocouple must agree to within 1.5% at each of the calibration
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points. A calibration curve (equation) may be constructed (calculated)
and the data extrapolated to cover the entire temperature range suggested
by the manufacturer.
8.6 Barometer - Adjust the barometer initially and before each test series
to agree to within ±2.5 mm Hg (0.1 in. Hg) of the mercury barometer or the
corrected barometric pressure value reported by a nearby National Weather Service
Station (same altitude above sea level).
8.7 Triple-beam or Electronic Balance - Calibrate the balance before each
test series, using Class S standard weights. The weights must be within ±0.5%
of the standards, or the balance must be adjusted to meet these limits.
9.0 CALCULATIONS
Perform calculations, retaining at least one extra decimal figure beyond
that of the acquired data. Round off figures after final calculations.
9.1 Total Formaldehyde - Determine the total formaldehyde in mg, using the
following equation:
[g/mole aldehyde]
Total mg formaldehyde = Cd x V x DF x x 103 mg/^ug
[g/mole DNPH derivative]
where:
Cd = measured concentration of DNPH-formaldehyde derivative, ng/ml
V = organic extract volume, mL
DF = dilution factor
9.2 Formaldehyde Concentration In Stack Gas - Determine the formaldehyde
concentration in the stack gas using the following equation:
Cf = K [total formaldehyde, mg] / Vm(std)
where:
K =35.31 ft3/")3 if Vm(std) is expressed in English units
= 1.00 m3/m3 if Vm(std, is expressed in metric units
Vm(stdi = volume of gas sample as measured by dry gas meter, corrected to
standard conditions, dscm (dscf)
9.3 Average Dry Gas Meter Temperature and Average Orifice Pressure Drop
are obtained from the data sheet.
9.4 Dry Gas Volume - Calculate Vm(std) and adjust for leakage, if necessary,
using the equation in Sec. 6.3 of EPA Method 5.
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9.5 Volume of Water Vapor and Moisture Content - Calculate the volume of
water vapor and moisture content from Equations 5-2 and 5-3 of EPA Method 5.
10.0 DETERMINATION OF VOLUME TO BE SAMPLED
To determine the minimum sample volume to be collected, use the following
sequence of equations.
10.1 From prior analysis of the waste feed, the concentration of
formaldehyde (FORM) introduced into the combustion system can be calculated. The
degree of destruction and removal efficiency that is required is used to
determine the maximum amount of FORM allowed to be present in the effluent. This
amount may be expressed as:
Max FORMi Mass = [(WF)(FORM, cone) (100 - %DRE)] / 100
where:
WF = mass flow rate of waste feed per h, g/h (Ib/h)
FORM, = concentration of FORM (wt %) introduced into the combustion
process
ORE = percent Destruction and Removal Efficiency required
Max FORM = mass flow rate (g/h [lb/h]) of FORM emitted from the combustion
sources
10.2 The average discharge concentration of the FORM in the effluent gas
is determined by comparing the Max FORM with the volumetric flow rate being
exhausted from the source. Volumetric flow rate data are available as a result
of preliminary EPA Method 1-4 determinations:
Max FORMi cone = [Max FORM, Mass] / DVeff(std)
where:
DVeff(stdi = volumetric flow rate of exhaust gas, dscm (dscf)
FORM; cone = anticipated concentration of the FORM in the exhaust gas
stream, g/dscm (Ib/dscf)
10.3 In making this calculation, it is recommended that a safety margin
of at least ten be included.
[LDLFORM x 10] / [FORMi cone] = Vtbc
where:
LDLFORM = detectable amount of FORM in entire sampling train
Vtbc = minimum dry standard volume to be collected at dry-gas
meter
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10.4 The following analytical detection limits and DNPH Reagent Capacity
(based on a total volume of 200 mL in two impingers) must also be considered in
determining a volume to be sampled.
11.0 QUALITY CONTROL
11.1 Sampling - See EPA Manual 600/4-77-027b for Method 5 quality control.
11.2 Analysis - The quality assurance program required for this method
includes the analysis of field and method blanks, procedure validations, analysis
of field spikes, and analysis of reagent checks. The assessment of combustion
data and positive identification and quantitation of formaldehyde are dependent
on the integrity of the samples received and the precision and accuracy of the
analytical methodology. Quality Assurance procedures for this method are
designed to monitor the performance of the analytical methodology and to provide
the required information to take corrective action if problems are observed in
laboratory operations or in field sampling activities.
11.2.1 Field Blanks - Field blanks must be submitted with the
samples collected at each sampling site. The field blanks include the
sample bottles containing aliquots of sample recovery solvents, methylene
chloride and water, and unused DNPH reagent. At a minimum, one complete
sampling train will be assembled in the field staging area, taken to the
sampling area, and leak-checked at the beginning and end of the testing
(or for the same total number of times as the actual sampling train). The
probe of the blank train must be heated during the sample test. The train
will be recovered as if it were an actual test sample. No gaseous sample
will be passed through the Blank sampling train.
11.2.2 Method Blanks - A method blank must be prepared for each set
of analytical operations, to evaluate contamination and artifacts that can
be derived from glassware, reagents, and sample handling in the
laboratory.
11.2.3 Field Spikes - A field spike is performed by introducing
200 nl of the Field Spike Standard into an impinger containing 200 mL of
DNPH solution. Standard impinger recovery procedures are followed and the
field spike sample is returned to the laboratory for analysis. The field
spike is used as a check on field handling and recovery procedures. An
aliquot of the field spike standard is retained in the laboratory for
derivatization and comparative analysis.
11.2.4 DNPH Reagent Checks - An aliquot of the extracted DNPH
reagent is prepared and analyzed according to the procedure in Sec. 5.4.3
to ensure that the background in the reagent is acceptable for field use.
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12.0 METHOD PERFORMANCE
12.1 Method performance evaluation - The following expected method
performance parameters for precision, accuracy, and detection limits are provided
in Table 3.
13.0 REFERENCES
1. U.S. Environmental Protection Agency, 40 CFR Part 60 Appendix A, Test
Methods.
2. Martin, R.M., "Construction Details of Isokinetic Source-Sampling
Equipment", U.S. Environmental Protection Agency, Research Triangle Park,
NC, Air Pollution Technical Document (APTD) 0581, April 1971.
3. Rom, J.J., "Maintenance, Calibration, and Operation of Isokinetic Source
Sampling Equipment", U.S. Environmental Protection Agency, Research
Triangle Park, NC, Air Pollution Technical Document (APTD) 0576, March
1972.
4. Annual Book of ASTM Standards. Part 26. Gaseous Fuels; Coal and Coke;
Atmospheric Analysis. American Society for Testing and Materials (ASTM),
Philadelphia, PA, 1974, pp. 617-622.
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TABLE 1
APPROXIMATE AMOUNT OF CRYSTALLINE DNPH USED
TO PREPARE A SATURATED SOLUTION
Amount of Moisture in DNPH Weight Required per 8 L of Solution
10 weight percent 31 g
15 weight percent 33 g
30 weight percent 40 g
TABLE 2
INSTRUMENT DETECTION LIMITS AND REAGENT CAPACITY
FOR FORMALDEHYDE ANALYSIS1
Analyte Detection Limit, ppbv2 Reagent Capacity, ppmv
Formaldehyde
Acetaldehyde
Acrolein
Acetone/Propi onal dehyde
Butyraldehyde
Methyl ethyl ketone
Valeral dehyde
Isovaleraldehyde
Hexal dehyde
Benzal dehyde
o-/m-/p-Tolualdehyde
Dimethyl benzal dehyde
1.8
1.7
1.5
1.5
1.5
1.5
1.5
1.4
1.3
1.4
1.3
1.2
66
70
75
75
79
79
84
84
88
84
89
93
1 Oxygenated compounds in addition to formaldehyde are included for comparison
with formaldehyde; extension of the methodology to other compounds is possible.
2 Detection limits are determined in solvent. These values therefore represent
the optimum capability of the methodology.
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TABLE 3
EXPECTED METHOD PERFORMANCE FOR FORMALDEHYDE
3
Parameter Precision1 Accuracy2 Detection Limit
Matrix: Dual ±15% RPD ±20% 1.4 x 10'10 lb/ft3
trains (1.8 ppbv)
1 Relative percent difference limit for dual trains.
2 Limit for field spike recoveries.
3 The lower reporting limit having less than 1% probability of false positive
detection.
0011 - 23 Revision 0
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APPENDIX A
ADDITION OF A FILTER TO THE FORMALDEHYDE SAMPLING TRAIN
As a check on the survival of participate material through the impinger
system, a filter can be added to the impinger train either after the second
impinger or after the third impinger. Since the impingers are in an ice bath,
there is no reason to heat the filter at this point.
Any suitable medium (e.g., paper, organic membrane) may be used for the
filter if the material conforms to the following specifications:
1) The filter has at least 95% collection efficiency (<5% penetration) for 3
/xm dioctyl phthalate smoke particles. The filter efficiency test shall
be conducted in accordance with ASTM standard method D2986-71. Test data
from the supplier's quality control program are sufficient for this
purpose.
2) The filter has a low aldehyde blank value (<0.015 mg formaldehyde/cm2 of
filter area). Before the test series, determine the average formaldehyde
blank value of at least three filters (from the lot to be used for
sampling) using the applicable analytical procedures.
Recover the exposed filter into a separate clean container and return the
container over ice to the laboratory for analysis. If the filter is being
analyzed for formaldehyde, the filter may be recovered into a container or DNPH
reagent for shipment back to the laboratory. If the filter is being examined for
the presence of particulate material, the filter may be recovered into a clean
dry container and returned to the laboratory.
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METHOD 0023A
SAMPLING METHOD FOR POLYCHLORINATED DIBENZO-p-DIOXINS
AND POLYCHLORINATED DIBENZOFURAN EMISSIONS
FROM STATIONARY SOURCES
1.0 SCOPE AND APPLICATION
1.1 This method describes the sampling procedure to be used for
determining stack emissions of polychlorinated dibenzo-£-dioxins (PCDDs) and
polychlorinated dibenzofurans (PCDFs) from stationary sources. The air sample
is collected and analyzed by the determinative portion of Methods 8280 or 8290
(Sees. 7.6 - 7.8). This method describes the procedures for sampling and
calculating results. This method may be modified to allow simultaneous sampling
and analysis for polychlorinated biphenyls (PCBs), polynuclear aromatic
hydrocarbons (PAHs), or semivolatile organic compounds (SVOCs). However,
specific approval is required for this modification, and detailed modification
of the methodology is required.
1.1.1 This method is a revision of EPA Air Method 23 (Ref. 12).
1.1.2 The surrogates and recovery standards include the standards
listed in Methods 8280 and 8290 (Table 1). However, analysts should add
the compounds only one time, during sampling.
1.1.3 The method refers to specific techniques described in EPA Air
Methods 1, 2 and 5. Analysts should obtain copies of those methods prior
to sampling.
1.2 This method is restricted to use by or under the supervision of
analysts experienced in the use of air sampling methods and the analysis of
PCDDs, PCDFs, PCBs, PAHs, and SVOCs from the components of Method 0010 trains.
Each analyst must demonstrate the ability to generate acceptable results with
this method.
1.3 Safety - The laboratory should develop a strict safety program for the
handling of PCDDs and/or PCDFs.
1.3.1 2,3,7,8-TCDD 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 must be aware 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. Personnel
handling these types of samples should wear masks fitted with charcoal
filters to prevent the inhalation of airborne particulates.
1.3.2 The toxicity or carcinogenicity of each reagent used in this
method is not precisely defined. However, each chemical should be treated
as a potential health hazard, and exposure to these chemicals kept to a
minimum. 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
0023A - 1 Revision 1
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should be made available to all personnel involved in the sampling and
chemical analysis of samples suspected to contain PCDDs/PCDFs. Additional
information on laboratory safety is given in References 4, 9, 10 and 11.
2.0 SUMMARY OF METHOD
2.1 Gaseous and particulate PCDDs/PCDFs are isokinetically withdrawn from
an emission source and collected in a multicomponent sampling train. The
collection components consist of the front half glassware surfaces (nozzle,
probe, and front half filter holder), the glass fiber filter, the back half
glassware surfaces (back half filter holder and condenser coil) and the solid
sorbent (XAD-2®) module.
2.2 Following sampling the glass collection components are rinsed. The
PCDD/PCDF are then extracted from the front half rinses and filter and another
separate extraction is performed on the XAD-2® and back half rinses.
2.3 The filter and XAD-2® extracts are then analyzed separately.
Surrogate recoveries are determined for both fractions. The analysis is
performed using high resolution gas chromatography (HRGC) and high resolution
mass spectrometry (HRMS), using the procedures of Method 8290.
3.0 INTERFERENCES
3.1 Polychlorinated biphenyls (PCBs) and polychlorinated diphenyl ethers
(PCDPEs) may interfere with low resolution gas chromatography/low resolution mass
spectroscopy PCDD/PCDF analyses, since these compounds produce many of the same
nominal masses as the PCDDs and PCDFs. However, high resolution mass
spectrometric techniques, in combination with capillary gas chromatography, are
typically used to resolve PCDD/PCDFs from these analytical interferences.
3.2 Very high amounts of other organic compounds in the matrix will
interfere with the analysis. Extensive column-chromatographic cleanup has been
introduced into typical HRGC/HRMS analytical methodology to minimize matrix
effects due to high concentrations of organic compounds.
3.3 Method 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 from interferences under the
conditions of the analysis by preparing and analyzing laboratory method blanks.
3.3.1 Glassware must be cleaned thoroughly before using. The
glassware should be washed with laboratory detergent in hot water followed
by rinsing with tap water and distilled water. The glassware may be
cleaned by baking in a glassware oven at 400°C for at least one hour.
After the glassware has cooled, the glassware should be rinsed three times
with methanol and three times with methylene chloride. Volumetric
glassware should not be heated to 400°C. Rather, after washing and
rinsing, volumetric glassware may be rinsed with methanol followed by
methylene chloride and allowed to dry in air.
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3.3.2 The use of high purity reagents and solvents helps to minimize
interference problems in sample analysis.
4.0 APPARATUS AND MATERIALS
The following section describes all the sampling equipment and the
associated performance specifications necessary to collect a gas sample from a
stationary source according to Method 0023.
4.1 Sampling train - A schematic diagram of the sampling train is shown
in Figure 1. This train configuration has been adapted from EPA Method 5 (40 CFR
Part 60 Appendix A) with the addition of condenser, XAD-2® trap and
filtration-coil connecting glassware. Sealing greases must not be used in
assembling the train. Complete sampling systems are commercially available that
have been developed to meet all the EPA equipment design specifications. The
following equipment is required.
4.1.1 Nozzle - The nozzle should be made of quartz or borosilicate
glass. Stainless steel nozzles should not be used. The taper angle should
be < 30°, with taper on the outside to preserve a constant inside diameter
(ID). The nozzle ID should be determined in order to sample isokinetically
at a rate that allows collection of an adequate sample volume. The minimum
sample volume should be determined to allow appropriate detection limits
to be achieved (see Sec. 9.0).
4.1.2 Probe liner - The sampling probe liner should be constructed
of borosilicate or quartz glass tubing. The typical outside diameter (OD)
used by sampling equipment manufacturers is about 16 mm, encased in a
stainless steel sheath with an OD of 25.4 mm. Either borosilicate or
quartz glass liners may be used for stack temperatures up to about 480°C,
but quartz glass liners should be used at higher stack temperature [480 to
900°C].
4.1.3 Probe sheath and heating system - A stainless steel or
equivalent probe sheath should be used to house the probe liner and heating
system. The probe heating system should be capable of maintaining probe
gas temperatures at the probe exit of 120°C ± 14°C during sampling. This
temperature should be verified by placing a thermocouple temperature sensor
against the outer surface of the probe liner at least 2 feet upstream of
the filter oven. Temperature readings should be recorded during sampling.
4.1.4 Glass cyclone - A glass cyclone may be used between the probe
and filter holder for high particulate concentrations. A cyclone, if used,
should be rinsed and recovered with the front half of the train.
4.1.5 Filter holder - A filter holder of borosilicate glass with a
Teflon® frit filter support should be used. The holder design should
provide a positive seal against leakage from the outside or around the
filter. The holder should be durable, easy to load, leak-free in normal
applications, and is positioned immediately following the probe (or
cyclone, if used) with the filter placed toward the flow.
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4.1.6 Filter heating system - Any heating system may be used which
is capable of maintaining the filter holder at 120°C ± 14°C during
sampling. Other temperatures may be specified by a subpart of the
regulations or approved for a particular application. A gauge capable of
measuring temperatures to within 3°C should be provided to monitor the
temperature around the filter during sampling.
4.1.7 Sample transfer lines - A sample transfer line may be used if
needed to direct sample flow from the probe to the filter or from the
filter to the condenser. The probe-to-filter line should be insulated and
heated so that gas exit temperatures are 120°C ± 14°C. The filter-to-
condenser line should be insulated and oriented with the downstream end
lower than the upstream end so that any condensate will flow away from the
filter and into the condenser. These lines should be constructed of
Teflon® or glass and should be recovered with their respective rinse
fractions (front half or back half).
4.1.8 Condenser - A multi-coil water-cooled glass condenser should
be used to cool the sample gas prior to entry into the sorbent module. The
orientation of the condenser should be vertical.
4.1.9 Sorbent module - The glass water-cooled container configured
to hold the solid sorbent (XAD-2®) should contain a minimum of 20 g of XAD-
2® and may contain as much as 40 g. A schematic diagram is shown in Figure
2. A single piece condenser-trap can be used if desired. The sorbent trap
configuration should be vertical so that condensate drains from the
condenser through the sorbent and so that channeling of the gas flow does
not occur. The connecting fittings should form leak-free, vacuum tight
seals. Sealant greases should not be used in the sampling train. A coarse
glass or Teflon® frit along with glass wool plugs is included to retain the
sorbent. The tester may engrave a unique identification number for
inventory and sample tracking.
4.1.10 Impinger trains - Four impingers should be connected in series
with leak-free ground-glass fittings or any similar noncontaminating
fittings. The first impinger should be a short stem (knock out) version.
The second impinger should be a Greenburg-Smith impinger with the standard
tip and plate. The third and fourth impingers should be the
Greenburg-Smith design modified so that the glass tube has an unconstricted
13 mm ID and extends to within 13 mm of the flask bottom. The fourth
impinger outlet connection should allow insertion of a thermometer capable
of measuring ± 1°C of true value in the range of 0 to 25°C.
4.1.11 Water circulating bath - A bath and pump circulating system
which is capable of providing chilled water flow to the condenser and
sorbent trap water jackets should be used. Typically a submersible pump
is placed in the impinger ice water bath so that the ice water contained
there can be used. The function of this system should be verified by
measuring sorbent trap gas entrance temperature
4.1.12 Pitot tube - The pitot tube, preferably of Type S design,
shall meet the requirements of EPA Method 2. The pitot tube is attached
to the probe as shown in Figure 1. The proper pitot tube-sampling nozzle
configuration for prevention of aerodynamic interference is shown in
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Figures 2.6 and 2.7 of EPA Method 2. The Type S pilot tube assembly shall
have a known coefficient, determined as outlined in Sec. 4 of EPA Method 2.
4.1.13 Differential pressure gauge - The differential pressure gauge
should be an inclined manometer or the equivalent as described in EPA
Method 2. Two gauges are required: one gauge to monitor the stack
velocity pressure (AP), and the other to measure the orifice pressure
differential (AH).
4.1.14 Metering system - The metering system should consist of a dry
gas meter with 2% accuracy, a vacuum pump, a vacuum gauge, orifice meter,
thermometers or thermocouples capable of measuring ± 3°C of true value in
the range of 0 to 90°C; and related equipment as shown in Figure 1.
Thermocouples should be used to monitor the temperature at the following
sampling train locations:
• stack gas
• probe liner
• filter holder
• sorbent trap entrance
• silica gel impinger exit
• dry gas meter inlet and
• dry gas meter outlet.
Other metering systems capable of maintaining isokinetic sampling rates
within 10% and determining sample volumes to within 2% may be used if
approved. Sampling trains with metering systems designed for sampling
rates higher than those described in APTD-0581 and APTD-0576 (Air Pollution
Technical Document, see references) may be used if the above specifications
can be met. When the metering system is used with a pitot tube, the system
should permit verification of an isokinetic sampling rate through the use
of a nomograph or by calculation.
4.1.15 Barometer - A mercury (Hg), aneroid, or other barometer
capable of measuring atmospheric pressure to within ± 2.5 mm Hg is needed.
A preliminary check of a new barometer should be made against a
mercury-in-glass barometer or the equivalent. The absolute barometric
pressure may be obtained from a nearby weather service station and adjusted
for elevation difference between the station and the sampling point.
Either subtract 2.5 mm Hg from the station value for every 30 m elevation
increase or add the same for an elevation decrease. If the barometer
cannot be adjusted to agree within 0.1 in. Hg of the reference barometric
pressure, it should be repaired or discarded.
4.1.16 Gas density determination equipment - The equipment necessary
for conducting EPA Methods 2-4 for determining stack gas flow, molecular
weight and moisture content, respectively, should be used. Required
measurements include stack gas velocity and static pressure; gas
temperature; concentrations of 02, C02, and N2 (by difference), metered gas
volumes and meter temperatures and pressure; and condensate weight gain
collected by the impinger train. All equipment should meet EPA Method 2 -
4 requirements.
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4.2 Sample recovery equipment
4.2.1 Fitting caps - Ground glass or cleaned aluminum foil to cap
the exposed sections of the train.
4.2.2 Wash bottles - Teflon®, 500-mL.
4.2.3 Probe-liner, probe-nozzle, and filter-holder brushes - These
should be constructed with nylon or Teflon® bristles with precleaned
stainless steel or Teflon® handles. The probe brush should have extensions
of stainless steel or Teflon® at least as long as the probe. The brushes
should be properly sized and shaped to brush out the nozzle, probe liner,
and front half filter holder.
4.2.4 Filter storage container - Typically a glass petri dish sealed
with Teflon® tape is used. Petri dishes should be cleaned according to
glassware cleaning procedures listed in this method (Sec. 6.1.4).
4.2.5 Balance - This balance is used for measuring weight gain of
the impingers and sample bottle weights as well. Typically a 0-2000-g
balance is used. The balance should be accurate to within 0.5 g, verified
with Class S weights.
4.2.6 Aluminum foil - Heavy duty cleaned by rinsing three times with
methylene chloride and once with toluene, stored in pre-cleaned glass petri
dish or glass jar.
4.2.7 Graduated cylinder - Glass, 250-mL, with ± 1 ml resolution
(this cylinder can be used for impinger volume determinations in place of
the balance).
4.2.8 Glass sample storage container - Amber glass bottle for sample
glassware washes, 500- or 1000-mL, with leak-free Teflon®-!ined caps. The
bottles should be either purchased as precleaned or cleaned according to
glassware cleaning procedures listed in this method (Sec. 6.1.4)
5.0 REAGENTS
5.1 Filters - Glass fiber filters, without organic binder, exhibiting at
least 99.95% efficiency (< 0.05% penetration) on 0.3 urn dioctyl phthalate smoke
particles. All filters should be cleaned before their initial use according to
the following procedures.
5.1.1 Precleaning - All filters should be cleaned before their
initial use. Place no more than 50 filters in a Soxhlet extraction
apparatus. Charge the Soxhlet with toluene and reflux for 16 hours. After
extraction, allow the Soxhlet to cool. Remove the filters and dry under
a clean nitrogen (N2) stream. Store the filters in cleaned glass petri
dishes or amber glass bottles sealed with Teflon® tape or Teflon®-! ined
caps prior to using them.
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5.1.2 As a quality control check prior to the field test, take one
precleaned filter and perform Soxhlet extraction with toluene for 16 hours.
Remove the toluene extract and analyze according to Method 8290. No
analytes may be observed above the detection limit.
5.1.3 Filter surrogate spike solution - As stated in Sec. 7.3.3,
Method 0023 calls for both the filter and the XAD-2® sorbent to be spiked
with the same set of isotopically labeled PCDD/PCDF standards. Surrogate
spikes are added to the sorbent prior to sampling and to the filter
immediately before the sample extraction. The filter and XAD-2® fractions
(including the associated glassware rinses) are extracted separately and
analyzed separately. The surrogate standards listed in Table 1 should be
used for both the filter spike and sorbent spike.
5.1.4 To ensure proper filter spiking, the isotopically-labeled
standard solution, which is normally at a concentration of 0.1 ng//il_, is
diluted to 0.004 ng//nL with nonane, for a dilution factor of 25. This
spiking solution will be used to spike the surface of the filter as
discussed in Sec. 7.3.1.
5.2 Sorbent resin - Amberlite XAD-2® resin. XAD-2® may be purchased
precleaned or cleaned by the laboratory. If the resin is not precleaned, the
cleaning procedures listed below shall be implemented.
5.2.1 Sorbent resin cleaning procedure
5.2.1.1 Place the sorbent resin in a clean beaker and rinse
with reagent water. Discard the rinse. Fill the beaker a second time
with reagent water and allow the resin to stand overnight. Discard
this second rinse.
5.2.1.2 Place the sorbent resin in an all-glass thimble of
a large Soxhlet extractor. The sorbent resin will float when in
contact with methylene chloride. Therefore, add a glass wool plug on
top of the resin in the thimble, and weight the glass wool plug down
with a stainless steel ring that fits inside the thimble.
5.2.1.3 Place the thimble filled with resin into the Soxhlet
extractor, add organic-free reagent water to the distilling flask,
apply heat, and extract the resin for 8 hours.
5.2.1.4 Allow the Soxhlet extractor to cool, discard the
water, and add methanol to the extractor. Apply heat and extract for
22 hours.
5.2.1.5 Again allowing the extractor to cool, drain off the
methanol, replace it with methylene chloride. Make sure that the
stainless steel ring and glass wool plug are still in place and
extract for 22 hours.
5.2.1.6 Extract the resin a fourth time, using toluene as
the extraction solvent, for 22 hours.
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5.2.1.7 Following the toluene extraction, the sorbent resin
must be dried under a stream of clean dry nitrogen or other inert gas.
This may be accomplished by transferring the resin to a large diameter
glass column and flowing the gas through the column. The gas may be
heated to less than 40"C, using a steam bath or other appropriate heat
source. Continue the inert gas flow through the resin until all the
residual solvent is removed. The flow rate should be sufficient to
agitate the resin particles, but not so excessive as to cause the
particles to fracture.
5.2.1.8 A quality control check should be conducted on the
cleaned sorbent using HRGC/HRMS techniques (Method 8290). Typically,
a method blank conducted previously on the same lot of sorbent can
serve this purpose.
5.2.2 Sorbent resin surrogate spike solution - The-XAD-2® sorbent
is spiked with isotopically labeled PCDD/PCDF standards prior to sampling
(surrogate spikes).
5.3 Glass wool - Cleaned by sequential immersion in three aliquots of
methylene chloride and one aliquot of toluene, dried in a 110°C oven, and stored
in a toluene-washed glass jar with a Teflon®-!ined screw cap.
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 Silica gel - Indicating type, 6 to 16 mesh. If previously used, dry
at 175°C for two hours. New silica gel may be used as received. Alternatively,
other types of desiccants may be used, provided that appropriate performance has
been demonstrated.
5.6 Recovery solvents - Solvents must be pesticide quality or equivalent.
5.6.1 Acetone, CH3COCH3
5.6.2 Methylene chloride, CH2C12
5.6.3 Toluene, C6H5CH3
6.0 SAMPLING COLLECTION, PRESERVATION, AND PREPARATION
This section addresses preparation and collection procedures for sampling.
6.1 Laboratory preparation
6.1.1 Filters. (See Sec. 5.1.)
6.1.2 Sorbent trap. (See Sec. 5.2.)
6.1.3 Glass wool - Precleaning and storage. (See Sec. 5.3)
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6.1.4 Glassware - All glass components of the train should be
cleaned as follows:
Soak all glassware in hot soapy water (Alconox® or equivalent).
Rinse with tap water to remove soap.
Rinse with disti11ed/deionized H20 (three times).
Bake at 400"C for 2 hours.
Rinse with methylene chloride (pesticide grade) (three times).
Rinse with toluene (pesticide grade) (three times).
Cap glassware with clean glass caps or cleaned aluminum foil.
Mark cleaned glassware with color-coded identification stickers.
Rinse glassware immediately before using with acetone and methylene
chloride.
6.1.5 Because probe liners do not usually fit in glassware baths or
ovens, they may be rinsed three times with methylene chloride followed by
three rinses with toluene, and sealed during transport.
6.2 Preliminary field determinations
6.2.1 Sample site - The sampling site and the minimum number of
sampling points should be selected according to EPA Method 1 or as
specified by the Agency. The stack static pressure, temperature, and range
of velocity pressures (APs) should be determined using EPA Method 2. The
stack gas moisture content should be determined using EPA Method 4, its
alternatives, previous data, or an engineering estimate. Stack gas 02 and
C02 concentrations should be estimated and dry molecular weight should be
calculated. These parameters are used to estimate the isokinetic sampling
rate settings.
6.2.2 Nozzle size - The nozzle size should be based on the range of
velocity pressures so that it is not necessary to change the nozzle size
in order to maintain isokinetic sampling rates.
6.2.3 Sampling duration - The total length of sampling time needed
to obtain the identified minimum sample gas volume is determined by
comparing the anticipated average sampling rate with the volume
requirement. (Average sampling rate should be within 0.5 to 0.75 cfm.)
The same time should be allocated to all traverse points defined by EPA
Method 1. To avoid timekeeping errors, the length of time sampled at each
traverse point should be an integer or an integer plus one-half minute.
6.2.3.1 Calculation of length of the sampling duration - The
minimum sampling time required to achieve a minimum sample volume and
the corresponding detection limit (DL) are given below.
... . TO. analytical DL
Minimum sample time =
(Sample Rate) x (desired gas cone. DL)
6.2.3.2 The following calculation is for a single isomer
(i.e., 2,3,7,8-TCDF). Detection limits for other isomers may need to
be calculated as well. For this example, it will be assumed that the
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analytical detection limit is 0.5 ng (actual analytical detection
limit will need to be specified for each test program).
6.2.3.3 At a sampling rate of 0.014 m3/min (0.5 cfm), the
sample volume per hour will be 0.85 m3/h. Assuming a desired stack
gas concentration detection limit to be 0.1 ng/m3, the minimum sample
time required to collect 0.5 ng at concentration in the stack of
0.1 ng/m would be:
Minimum sample time = : = 6.25 h
0.85 m3/h x 0.1 ng/m3
6.2.3.4 The total sampling time should be greater than or
equal to the minimum total sampling time required to achieve the
necessary detection limit. In addition, the sampling time per point
should be greater than 2 min (greater minimum time interval may be
specified by the Agency), and the sample volume corrected to standard
conditions shall exceed the required minimum total gas sample volume.
6.3 Calibration
Calibration of the apparatus is one of the most important functions in
maintaining data quality. The detailed calibration procedures for the sampling
apparatus listed in this section can be found in EPA Method 5 and Method 0010.
Table 4 summarizes the quality assurance functions for the calibrations.
6.3.1 Metering system
6.3.1.1 Full dry gas meter calibration - The dry gas meter
(DGM) in the meter console of the sampling system should be fully
calibrated against a primary standard meter (wet test meter or
spirometer) or alternatively against a second reference meter (dry gas
meter or critical orifice) that has been calibrated against a primary
standard meter. The procedure can be found in Sees. 5.3 and 5.7 of
EPA Method 5.
6.3.1.2 Post-test DGM calibration check - Following the test
program, the full calibration factor or meter Y should be checked by
performing a post-test DGM calibration check. Any secondary reference
meters can be used. Three calibration runs are conducted at the
maximum vacuum reached during the testing. The average post-test
calibration factor should not deviate from the full DGM calibration
factor by more than 5%. Additional details on these procedures can
be found in Sec. 5.3.2 of EPA Method 5.
6.3.2 Temperature gauges - Each thermocouple should be permanently
and uniquely marked on the casting; all mercury-in-glass reference
thermometers should conform to ASTM E-l 63C or 63F specifications.
Thermocouples should be calibrated in the laboratory with and without the
use of extension leads. If extension leads are used in the field, the
thermocouple readings at ambient air temperatures, with and without the
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extension lead, should be noted and recorded. Correction is necessary if
the use of an extension lead produces a change greater than 1.5 percent.
6.3.2.1 Impinger, organic module, and dry gas meter
thermocouples - For the thermocouples used to measure the temperature
of the gas leaving the impinger train and the XAD-2® resin bed,
three-point calibration at ice-water, room-air, and boiling-water
temperatures is necessary. The thermocouples should be accepted only
if the readings at all three temperatures agree to ± 2°C with those
of the absolute value of the reference thermometer.
6.3.2.2 Probe and stack thermocouple - For the thermocouples
used to indicate the probe and stack temperatures, a three-point
calibration at ice-water, boiling-water, and hot-oil-bath temperatures
should be performed; it is recommended that room-air temperature be
added, and that the thermometer and the thermocouple agree to within
1.5% at each of the calibration points. A calibration curve
(equation) may be constructed and the data extrapolated to cover the
entire temperature range suggested by the manufacturer.
6.3.3 Probe heater - The probe heating system should be calibrated
prior to field use according to the procedure outlined in APTD-0576.
Probes constructed according to APTD-0581 need not be calibrated if the
curves of APTD-0576 are used.
6.3.4 Barometer - The field barometer should be adjusted initially
and before each test series to agree within 2.5 mm Hg of the
mercury-in-glass barometer or with the station pressure value reported by
a nearby National Weather Service station, corrected for elevation. The
correction for elevation difference between the station and the sampling
point should be applied at a rate of -2.4 mm Hg/30 m of elevation increase.
The results should be recorded on the pretest sampling check form.
6.3.5 Probe nozzle - Probe nozzles should be calibrated before
initial use in the field. The ID of the nozzle should be measured with a
micrometer to the nearest 0.025 mm. Three measurements should be made
using different diameters each time and the average obtained. The
difference between the high and the low numbers should not exceed 0.1 mm.
When nozzles become damaged they should not be used again. Each nozzle
should be permanently and uniquely identified.
6.3.6 Pitot tube - The Type S pitot tube assembly should be
calibrated using the procedure outlined in EPA Method 2.
6.3.7 Balance - The balance should be calibrated initially by using
Class-S standard weights and should be within 0.5 g of the standard weight.
6.4 Sampling train preparation - Care should be taken to ensure a clean
sampling train preparation area free of excessive dust and organic compounds for
preparing the sampling train.
6.4.1 Preparation of impingers - During preparation and assembly of
the sampling train, all train openings where contamination can enter should
be sealed until just prior to assembly or until sampling is about to begin.
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6.4.1.1 The first impinger should be left empty (used as a
water knock-out impinger due to long run times).
6.4.1.2 Approximately 100 ml of reagent water should be
placed in the second and third impingers. This method does not
require that organic analyses be conducted on the impinger contents.
However, if analyses of semivolatile organic compounds are to be
conducted, then the proper specifications on cleaning the impingers
and water quality (i.e., HPLC-grade water) should be observed.
6.4.1.3 Approximately 200 to 300 g of silica gel should be
placed in the fourth impinger. All impingers should be weighed
separately to the nearest 0.5 g and the weights recorded. Impingers
should be connected with glass U-tube connectors.
6.4.2 Filter loading - A filter should be placed in a
properly-cleaned filter holder using cleaned tweezers or clean disposable
surgical gloves. The filter should be properly centered and the gasket (if
used) properly placed to prevent the sample gas stream from circumventing
the filter. The filter should be checked for tears after the assembly is
completed.
6.4.3 Sorbent loading - The XAD-2® should be loaded and sealed in
the analytical (preparation) laboratory.
6.4.4 Final assembly - The final assembly of the filter holder,
condenser, and sorbent module can be performed at the stack location. All
components should be sealed with either precleaned foil or socket joints.
6.5 Sampling train leak check procedures - Leak checks are necessary to
assure that the sample has not been biased low by dilution air. Both pre-test
and post-test leak checks are necessary.
6.5.1 Pre-test - After the sampling train has been assembled, the
train should be leak checked at the sampling site by plugging the nozzle
and pulling a 380 mm Hg vacuum. Leakage rates greater than 4% of the
average sampling rate or 0.00057 m3/min, whichever is less, are
unacceptable. Leak checks should be conducted according to EPA Method 5
criteria.
6.5.2 During the sampling - If a component (e.g., filter assembly,
sorbent module, or impinger) change is necessary during the sampling run,
a leak check should be conducted before the change. The leak check should
be done according to the procedure outlined above, except that it should
be at a vacuum equal to or greater than the maximum value recorded up to
that point in the test. If the leakage is less than 0.00057 m3/min or 4%
of the average sampling rate (whichever is less), the results are
acceptable. If, however, a higher leakage rate is obtained, the tester
should record the leakage rate and either void the sampling run or perform
sample volume leak corrections (if approved by the Agency). After
replacing the train component, an initial leak check should be completed
before sampling.
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6.5.3 Post-test - The leak check should be completed at a vacuum
equal to or greater than the maximum value reached during the sampling run.
If the leakage rate is less than 0.00057 m3/nrin or 4% of the average
sampling rate (whichever is less), the results are acceptable. If,
however, a higher leakage rate is obtained the tester shall either void the
sample run or perform sample volume leak corrections (if approved by the
Agency).
6.6 Sampling train operation
6.6.1 Final pre-test sampling checks - After conducting the initial
leak check, the following checks should be made:
• Meter box examination
• Manometers leveled and zeroed
• Pump checked for proper operation
• Pitot lines leak checked
• Probe markings verified
• Thermocouples reading correctly
• Size and orientation of the nozzle verified
• EPA Method 3 equipment for C02/02 checked for proper assembly and leak
checked and
• Isokinetic K-factor checked to ensure that it is correct.
Immediately prior to sampling:
• Portholes should be cleaned to minimize the chance of sampling
deposited material
• Probe and filter heating system temperatures should be checked
• Condenser/sorbent cooling system temperatures should be checked and
• Proper nozzle location should be verified.
6.6.2 The sampling procedure below should be followed.
6.6.2.1 Sampling - Initial dry gas meter readings,
barometric pressure, and temperatures should be recorded. The tip of
the probe should be positioned at the first sampling point with the
nozzle tip pointing directly into the gas stream. When the probe is
in position, the open area around the probe and the porthole should
be blocked off to prevent flow disturbances and non-representative
dilution of the gas stream. The pump should be turned on and the
sample flow adjusted immediately to attain isokinetic conditions. The
EPA Method 3 sampling system should be turned on. Velocity pressures
should be recorded and the sampling rate adjusted to isokinetic.
Other readings of velocity pressure (AP), orifice pressure (AH), stack
gas temperature (TJ, probe temperature (Tp), filter temperature (Tf),
sorbent trap temperature (Tt), silica gel impinger temperature (Tsg),
dry gas meter inlet and outlet temperatures (Tm), dry gas meter
volume, and sample vacuum should be made.
6.6.2.2 The stack should be traversed as directed in EPA
Method 5 procedures. At each sample point, the above readings should
be taken and sample flow rates adjusted to isokinetic. Following the
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traverses, the pump is turned off, the probe removed from the stack,
and the final DGM readings recorded. Care should be taken not to bump
the nozzle against stack walls in order to minimize the chance of
breakage or extracting deposited material. Following each port
traverse, a leak check is recommended in order to ensure a leak tight
system. An additional leak check may also be performed after the
train is moved to the next port, prior to sampling. The necessary
post-test leak check should be conducted and the leak rate recorded.
6.6.2.3 Periodically during the test run, the connecting
glassware from the probe, through the filter, and to the condenser
should be checked for water condensation. If any condensation is
evident, verify that the temperature sensors and heater systems are
functioning properly. Ice should be maintained around the impingers
to keep both the sorbent trap entrance and silica gel exit temperature
at 20°C. Filter vacuum should be checked for sudden increases. The
filter should be changed if the vacuum exceeds 15 in. Hg. The
manometer level and zero should also be checked periodically during
each traverse, because vibrations and temperature fluctuations can
cause the manometer zero to shift.
6.6.2.4 Following the post-test leak check, the probe should
be disconnected, and the nozzle and the end of the probe capped with
precleaned aluminum foil, or equivalent caps. The inlet to the filter
holder should be capped according to one of the methods previously
mentioned. It may be necessary to loosen the seal between the sorbent
module outlet and the inlet to the first impinger to prevent water
from being drawn back into the module when the sample train cools.
Alternatively, the filter holder, condenser and sorbent module may be
disassembled and immediately capped at the stack location and removed
to the sample recovery area.
6.7 Collection of blanks - Four different sampling blanks should be
collected: field blanks, reagent blanks, proof blanks, and method blanks
(laboratory only). Only two sampling blanks should be analyzed initially: the
field blank and the laboratory method blank. If the field blank has high levels
of contamination and the laboratory blank does not show high background levels
of PCDD/PCDF, the other blanks should be analyzed to help determine the source
of the contamination. Blanks are further discussed in Sec. 8.0.
7.0 PROCEDURE
7.1 Recovery preparation - Proper recovery procedure begins as soon as the
probe is removed from the stack at the end of the sampling period. The nozzle
end of the sampling probe should be sealed with precleaned aluminum foil and
disconnected from the filter holder. When the probe is cool enough to be handled
safely, all external particulate matter near the tip of the probe should be wiped
off and both ends of the probe closed off with aluminum foil. Both openings to
the filter holder, transfer line (if used), condenser, sorbent trap, and impinger
train should be disconnected and sealed. Care should be taken not to lose any
condensed water upstream of the impingers (if present) during this process.
0023A - 14 Revision 1
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Train components should be transferred to the cleanup area. This area
should be clean and enclosed so that the chances of losing or contaminating the
sample are minimized. Smoking, which could contaminate the sample, is not
allowed in the cleanup area. Cleanup personnel should wash their hands prior to
sample recovery. The train should be inspected prior to and during disassembly
and any abnormal conditions, e.g., broken filter, colored impinger liquid, etc.,
noted.
7.2 Sample recovery procedure - As shown in Figure 3, the sampling train
should be recovered into four containers. The procedures applicable to each
sample container are briefly discussed in the following section.
7.2.1 Filter (Container 1) - The filter should be removed carefully
from the filter holder and placed in its identified container. Cleaned
tweezers should be used to handle the filter. Fold the filter, if
necessary, with the particulate cake inside the fold. Any particulate
matter and filter fibers which adhere to the filter holder gasket should
be transferred to the container by using a dry inert bristle brush and a
sharp-edged blade. The container should be sealed with Teflon® tape.
7.2.2 Front half rinse (Container 2) - The front half glassware
surfaces will be rinsed with acetone and brushed, followed by three
additional rinses with methylene chloride. All rinses should be put into
Container 2. The outside of the probe, the pitot tube, and the nozzle
should be cleaned to prevent particulates from being brushed into the
sample bottle. The probe liner should be tilted and rotated while
squirting acetone into the upper end to assure complete wetting of the
inside surface. Acetone is then squirted into the upper end while pushing
the probe brush through the liner with a twisting motion, with the drainage
caught in the sample bottle (Container 2). The brushing procedure should
be repeated two more times or until no particles are visible in the
drainage and a visual inspection of the liner reveals no particles
remaining inside. The brush should be rinsed into the sample bottle to
collect any particulates that may be retained within the bristles. The
three rinses should be repeated with methylene chloride allowing the
rinsate to collect into the same sample container.
After all the rinsings have been collected, the lid on the sample
container should be tightened securely. As a precaution in case of
leakage, the liquid level should be marked on the sample container and the
cap sealed with Teflon® tape. The sample recovery should be recorded on
the sample recovery form.
7.2.3 Sorbent module (Container 3) - The sorbent module should be
removed from the train, tightly capped at both ends with aluminum foil or
glass caps, labeled and stored on ice for transport to the laboratory.
Care should be taken to ensure that no ice water can leak into the stored
traps or any other train component.
7.2.4 Back half rinse (Container 4) - The back half of the filter
holder, transfer line (if used) and condenser (if separate from trap)
should be rinsed three times with acetone followed by three rinses with
methylene chloride. The sample container (Container 4) is then identified
and sealed as discussed above.
0023A - 15 Revision 1
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7.2.5 Impinger water - Any color or film in the impinger water
should be noted on the sample recovery form. The entrained moisture in the
first three impingers should be measured to within ± 1 mL by using a
graduated cylinder or by weighing to within 0.5 g by using a balance, and
the data recorded appropriately. This information is needed to calculate
the moisture content of the effluent gas.
7.2.6 Silica gel - The color of the indicating silica gel should be
noted on the recovery form to determine if it has been completely spent and
the impinger weighed to determine entrained moisture weight gain. Analysis
is not required.
7.3 Analysis summary - The following section summarizes the analytical
procedures for quantitating PCDD/PCDF collected by the sampling train. Sample
preparation procedures and the basic analytical techniques are listed. The
detailed analytical protocol can be found in Method 8290.
7.3.1 As shown in Figure 4, the analytical procedure requires the
sampling train to be analyzed in two fractions. Containers 1 and 2 (filter
and front half rinse) are combined and analyzed. Containers 3 and 4
(sorbent trap and back half rinse) are also combined and analyzed. In this
way filter surrogate standard recoveries and XAD-2® surrogate standard
recoveries are both determined separately.
7.3.2 Acceptance criteria and corrective actions for surrogate
recoveries are as follows:
7.3.2.1 All PCDD/PCDF surrogate recoveries should be within
70 to 130 percent.
7.3.2.2 If all isomer recoveries are greater than
130 percent, the sampling runs should be repeated,
7.3.2.3 If all isomer recoveries are less than 70 percent,
the sampling runs should either be repeated or the final results
should be divided by the fraction of surrogate recovery.
7.3.2.4 Acceptance criteria for other standard recoveries
(i.e., internal) should conform to Method 8290 requirements.
7.3.3 As discussed in Sees. 5.1.2 and 5.2.1, surrogate spikes are
added to the sorbent trap prior to sampling and to the filter immediately
prior to extraction. The same set of isotopically-labeled compounds is
used for these spikes. The analytical procedure for both fractions is
given in the following sections. All samples should be extracted within
30 days of collection and analyzed within 45 days of extraction.
7.3.4 Sample preparation and internal standard addition - The
following procedure should be performed for the filter/front half analysis
and the sorbent trap/back half analysis. The only difference between the
two procedures is that surrogate standards are added to the filter/front
half fraction immediately prior to sample preparation whereas the surrogate
standards have already been added to the sorbent trap/back half prior to
sampling.
0023A - 16 Revision 1
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7.3.4.1 Filter/front half fraction procedures - Place a
cellulose extraction thimble, 1 g of silica gel or sodium sulfate, and
a plug of glass wool into the Soxhlet apparatus, charge the apparatus
with toluene, and reflux for a minimum of 3 hours. Remove the toluene
and discard it, but retain the silica gel. Remove the extraction
thimble from the extraction system and place it in a glass beaker to
catch the solvent rinses.
7.3.4.2 Add exactly 1.0 ml of the surrogate spiking solution
(Sec. 5.1.2) uniformly onto the surface of the filter while it is
still in the petri dish in which it was returned from the field, using
an adjustable pipet. Transfer the filter directly to the extraction
thimble of the extraction system. Rinse the petri dish with 10 ml of
toluene three times collecting the rinsate into the beaker.
7.3.4.3 Concentrate the sample in Container 2 (acetone/
methylene chloride rinses) to a volume of about 1-2 ml_ using a
Kuderna-Danish concentrator apparatus, followed by nitrogen blow down
at a temperature of less than 37°C. Rinse the sample container three
times with small portions of methylene chloride and add these to the
concentrated solution and concentrate further to near dryness. This
residue contains particulate matter removed in the rinse of the train
probe and nozzle. Add the concentrate to the filter in the Soxhlet
apparatus described above.
7.3.4.4 Add 40 juL of the internal standard solution.
Fortification is accomplished by using the sample fortification
solutions described in Table 1. Cover the contents of the extraction
thimble with the cleaned glass wool plug and proceed to the extraction
procedure.
7.3.4.5 Sorbent trap/back half fraction procedures - Prepare
another extraction thimble/silica gel system as described above.
Suspend the adsorbent module directly over the extraction thimble in
the beaker. The glass frit of the module should be in the up
position. Using a Teflon® squeeze bottle containing toluene, flush
the XAD-2® into the thimble onto the bed of cleaned silica gel.
Thoroughly rinse the glass module catching the rinsings in the beaker
containing the thimble, first with methanol, if needed, then with
toluene into the thimble. If the resin is wet, effective extraction
can be accomplished by loosely packing the resin in the thimble. Add
glass wool plug from the XAD-2® sampling module to the thimble.
7.3.4.6 Concentrate the sample in Container 4
(acetone/methylene chloride rinses) to a volume of about 1 - 2 ml
using a Kuderna-Danish concentrator apparatus, followed by nitrogen
evaporation at a less than 37°C. Rinse the sample container three
times with small portions of methylene chloride and add these to the
concentrated solution and concentrate further to near dryness. Add
the concentrate to the XAD-2® resin in the Soxhlet apparatus described
above.
7.3.4.7 Add 40 /nL of the internal standard solution.
Fortification is accomplished by using the sample fortification
0023A - 17 Revision 1
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solutions described in Table 1. Cover the contents of the extraction
thimble with a cleaned glass wool plug to prevent the XAD-2® resin
from floating into the solvent reservoir of the extractor and proceed
with extraction (Sec. 7.3.2).
7.3.5 Sample extraction - Place the thimble in the extractor and add
the toluene contained in the beaker to the solvent reservoir. Pour
additional toluene to fill the reservoir approximately two-thirds full.
Add Teflon® boiling chips and assemble the apparatus. Adjust the heat
source to cause the extractor to cycle three times per hour. Extract the
sample for 16 hours. After extraction, allow the Soxhlet to cool. Transfer
the toluene extract and three 10-mL between rinses to the rotary
evaporator. Concentrate the extract to approximately 10 ml.
Use a nitrogen evaporative concentrator to reduce the volume of the
extract to about 100 juL. Redissolve the residue in 5 mL of hexane.
7.3.6 Sample clean-up and fractionation - Sample extracts described
above are spiked with 40 pi of the alternate standard fortification
solution, then divided into two equal portions. One half of each sample
extract is archived for future needs. The other portion is solvent-
exchanged to hexane then subjected to three column chromatographic cleanup
steps as described in Method 8290.
7.3.7 Analysis summary - The samples are analyzed with a high
resolution gas chromatographic column coupled to a high resolution mass
spectrometer (HRGC/HRMS) using the instrumental parameters described below.
Prior to analysis, the Recovery Standard solution from Table 1 is added to
each sample. Sample extracts are first analyzed using a capillary column
to determine the concentration of each isomer of PCDDs and PCDFs (tetra-
through octa-). If 2,3,7,8-TCDF is detected in this analysis, another
aliquot of the sample is analyzed separately, using a second, dissimilar
column to confirm and more accurately measure the 2,3,7,8-TCDF isomer.
Other column systems may be used, provided that the user is able to
demonstrate by means of calibration and performance checks that the column
system is able to meet the specifications of Sec. 5.6 of Method 8290.
7.3.8 All other analytical specifications for determining the
amounts of PCDD/PCDF isomers collected in the filter/front half and sorbent
trap/back half fractions can be found in Method 8290.
7.4 Calculations
The following section describes the calculations used to determine gas
concentrations and emissions of PCDD and PCDF isomers. Toxic equivalent
calculations are not included in this method. Each set of calculations should
be repeated or spot-checked, as a QC measure. Calculations should be carried out
to at least one extra decimal place beyond that of the acquired data and should
be rounded off after final calculation to two significant digits for each run or
sample. All rounding of numbers should be performed in accordance with the
ASTM 380-76 procedures.
The nomenclature and sampling equations are presented in Sec. 7.4.1.
0023A - 18 Revision 1
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7.4.1 Sampling nomenclature
An = Cross sectional area of nozzle, m2 (ft2).
As = Cross sectional area of stack, m2 (ft2).
BW8 = Water vapor in the gas stream, proportion by volume.
C; = Concentration of pollutant i, jug/dscm (Ib/dscf).
E| = Emission rate of pollutant i, g/sec (Ib/hr).
DN = Diameter of nozzle, mm (in.)
I = Percent of isokinetic sampling.
Mw = Molecular weight of water, 18.0 g/g-mole (18.0 Ib/lb-mole).
Md = Molecular weight of dry stack gas, g/g-mole (Ib/lb-mole).
M8 = Molecular weight of wet stack gas, g/g-mole (Ib/lb-mole).
ttij = Mass of pollutant i collected by sampling train, /ng (lb).
Pbar = Barometric pressure at the sampling site, mm Hg (in. Hg).
Static = Static gauge pressure of stack gas, mm H20 (in. H20).
P8 = Absolute stack gas pressure, mm Hg (in. Hg).
Pstd = Standard absolute pressure, 760 mm Hg (29.92 in. Hg).
Qsd = Average stack gas volumetric flow, dry, standard conditions,
dscmm (dscfm).
R = Ideal gas constant, 0.06236 [(mm Hg) (m3)] / [(°K) (g-mole)]
(21.85 [(in. Hg) (ft3)] / [(°R) (lb-mole)]}.
Tm = Absolute average DGM temperature°K (°R).
Ts = Absolute average stack gas temperature'K (°R).
Tstd = Standard absolute temperature, 293°K (528°R).
V,c = Total volume liquid collected in impingers and silica gel
(ml).
Vm = Volume of gas sample as measured by dry gas meter, dcm
(dcf).
= Volume of gas sample measured by the dry gas meter,
corrected to standard conditions, dscm (dscf).
0023A - 19 Revision 1
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Vw
-------
7.4.4 Moisture content
V
B =
w(std)
V + V
m(std) w(std)
NOTE: In saturated or water droplet-laden gas streams, two calculations of
the moisture content of the stack gas should be made, one from the
impinger analysis (Sec. 7.4.3), and a second from the assumption of
saturated conditions. The lower of the two values of Bws should be
considered correct. The procedure for determining the moisture
content based upon assumption of saturated conditions is given in
the Note in Sec. 1.2 of EPA Method 4. For the purposes of this
method, the average stack gas temperature may be used to make this
determination, provided that the accuracy of the in-stack
temperature sensor is ± 2°C.
7.4.5 Absolute stack gas pressure
P .
p _ p + static
bar 13.6
7.4.6 Average molecular weight of dry stack gas
Dry: Md = (0.32 x %02) x (0.44 x %C02) +(0.28 x (100 - (%02 + %C02))
Wet: M - M. x (1 - B ) + (B x M )
s d v ws' v ws w'
7.4.7 Stack gas velocity at stack conditions
Vs = K x C x
* T
std
P x M
7.4.8 Average stack gas volumetric flow at dry, standard conditions
d X P* 60sec
Qd = V xA x (1 -B ) x
^sd s s x ws '
T x P . min
s std
7.4.9 Concentration of pollutant
M.
C. = —'-
0023A - 21 Revision 1
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7.4.10 Emission of pollutant
7.4.11 Isokinetic sampling rate
1039.5746 x V x (T + 460)
V, X0x Ps x (1 -BwJ x (DJ2
*English units
8.0 QUALITY CONTROL
The following quality control (QC) guidelines outline pertinent steps to
be followed during the production of emission data to ensure and quantify the
acceptability and reliability of the data generated.
8.1 Sampling QC procedures - Quality control procedures specific to manual
source gas sampling procedures should follow EPA Method 5 and those listed in EPA
Manual 600/4-77-0276 for Method 5. Sampling QC procedures are summarized in
Table 2.
8.2 Blanks
8.2.1 Field blank - A field blank should be collected from a set of
glassware that has not been used to collect any field samples. The field
blank train is loaded, leak checked, and left at a sampling location during
a test run. The train is then recovered. The purpose of the field blank
is to measure the level of contamination that occurs from handling,
loading, recovering, and transporting the sampling train. Collect one
field blank for every nine test runs at each test location.
8.2.2 Glassware blank (proof blank) - A proof blank is recovered
from each set of sampling train glassware that is used to collect the
organic samples. The precleaned glassware, which consists of a probe
liner, filter holder, condenser coil, and impinger set, is loaded as if for
sampling and then quantitatively recovered exactly as the samples will be.
Analysis of the generated fractions will be performed to check the
effectiveness of the glassware cleaning procedure, but only if field blank
analysis indicates a potential contamination problem. If requested by the
test administrator, collect one glassware blank per each set of glassware
used on the complianca test and archive for future analysis in the event
the field blank shows contamination.
8.2.3 Reagent blank - Reagent blanks should contain 500 mL of each
reagent used at the test site. Reagent blanks are saved for potential
analysis. Each reagent blank is part of the same lot used during the
sampling program. If a field blank is unsatisfactory because of
contamination, reagent blanks may be analyzed to determine the specific
0023A - 22 Revision 1
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source of contamination. Collect one reagent blank per compliance test and
archive for future analysis in the event that the field blank shows
contamination.
8.2.4 Laboratory method blank - A method blank is a performance
control sample that is prepared in the laboratory and processed in a manner
identical to a field sample. The XAD-2® resin should be from the same
batch used for preparation of the field traps. One laboratory method blank
should be analyzed for every batch of samples analyzed.
9.0 METHOD PERFORMANCE
9.1 Method performance evaluation - Evaluation of analytical procedures
for a selected series of compounds shall include the sample preparation
procedures and each associated analytical determination. The analytical
procedures should be challenged by the test compounds spiked at appropriate
levels and carried through the procedures.
9.2 Method detection limit - The overall method detection limits (lower
and upper) should be calculated as shown in Sec. 6.2.3.1. Generally, analytical
detection limit for tetra-CDD/CDF congeners are 50 pg. Penta-, hexa-, and hepta-
congener detection limits are 250 pg and octa-congener detection limits are
500 pg.
9.3 Method precision and bias - The overall method precision and bias
should be determined on a compound-by-compound basis at a given concentration
level. The method precision value includes a combined variability due to
sampling, sample preparation, and instrumental analysis. The method bias is
dependent upon the collection, retention, and extraction efficiency of the train
components. Interlaboratory testing of Method 0023 and Method 8290 to establish
method accuracy and precision for sampling a variety of stationary sources has
not been performed.
10.0 REFERENCES
1. American Society of Mechanical Engineers, Sampling for the Determination
of Chlorinated Organic Compounds in Stack Emissions. Prepared for U.S.
Department of Energy and U.S. Environmental Protection Agency. Washington,
DC. December 1984.
2. American Society of Mechanical Engineers. Analytical Procedures to Assay
Stack Effluent Samples and Residual Combustion Products for Polychlorinated
Dibenzo-p-Dioxins (PCDD) and Polychlorinated Dibenzofurans (PCDF).
Prepared for the U.S. Department of Energy and U.S. Environmental
Protection Agency. Washington, DC. December 1984.
3. Thompson, J.R., Analysis of Pesticide Residues in Human and Environmental
Samples, U.S. Environmental Protection Agency, Research Triangle Park, NC,
1974.
4. U.S. Environmental Protection Agency. Method 8290: The Analysis of
Polychlorinated Dibenzo-p-Dioxins and Polychlorinated Dibenzofurans by
0023A - 23 Revision 1
January 1995
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High-Resolution Gas Chromatography/High-Resolution Mass Spectrometry. In:
Test Methods for Evaluating Solid Waste, SW-846. Washington, DC.
5. U.S. Environmental Protection Agency. Method 8280: The Analysis of
Polychlorinated Dibenzo-p-Dioxins and Polychlorinated Dibenzofurans. In:
Test Methods for Evaluating Solid Waste, SW-846. Washington, DC.
6. Tondeur, Y., Albro, P.W., Mass, R.J., Harvan, D.J., Schroeder, J.L.,
"Matrix Effect in Determination of 2,3,7,8-Tetrachlorodibenzodioxin by Mass
Spectrometry", Anal. Chem. 56(8), pp 1344-1347, 1984.
7. Tondeur, Y., Niederhut, W.N., Campana, J.E., Missler, S.R., "A Hybrid
HRGC/MS/MS Method for the Characterization of Tetrachlorinated-p-Dioxins
in Environmental Samples", Biomed. Environ. Mass Spectrom. 14(8), pp 449-
456, 1987.
8. Taylor, J.K., Quality Assurance of Chemical Measurements, Lewis Publishers,
Inc., 1987.
9. Department of Health, Education, and Welfare, Public Health Service, Center
for Disease Control. Carcinogens - Working with Carcinogens. Publication
No. 77-206. National Institute for Occupational Safety and Health. August
1977.
10. OSHA Safety and Health Standards, General Industry. 29 CFR, p 1910.
Occupational Safety and Health Administration. OSHA 2206. Revised January
1976.
11. American Chemical Society, Committee on Chemical Safety. Safety in
Academic Chemistry Laboratories. 3rd Edition, 1979.
12. 40 CFR Part 60, Appendix A.
13. Martin, R.M., Construction Details of Isokinetic Source-Sampling Equipment.
U. S. Environmental Protection Agency, Research Triangle Park, NC. Air
Pollution Technical Document (APTD) 0581, April 1971.
14. Rom, J.J., Maintenance, Calibration, and Operation of Isokinetic Source
Sampling Equipment. U.S. Environmental Protection Agency, Research
Triangle Park, NC. Air Pollution Technical Document (APTD) 0576, March
1972.
0023A - 24 Revision 1
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TABLE 1
COMPOSITION OF THE SAMPLE FORTIFICATION
AND RECOVERY STANDARDS SOLUTIONS
Analyte
Internal Standards
13C12-2,3,7,8-TCDD
13C12-l,2,3,7,8-PeCDD
13C12-l,2,3,6,7,8-HxCDD
13C12-l,2,3,4,6,7,8-HpCDD
13C12-OCDD
13C12-2,3,7,8-TCDF
13C12-l,2,3,7,8-PeCDF
13C12-l,2,3,6,7,8-HxCDF
13C12-l,2,3,4,6,7,8-HpCDF
Surrogate Standards
37Cl4-2,3,7,8-TCDD
13C12-l,2,3,4,7,8-HxCDD
13C12-2,3,4,7,8-PeCDF
13C12-l,2,3,4,7,8-HxCDF
13C12-l,2,3,4,7,8,9-HpCDF
Recovery Standards
13C12-1,2,3,4-TCDD
13C12-l,2,3,7,8,9-HxCDD
Alternate Standard
13C12-l,2,3,7,8,9-HxCDF
0023
Concentration
(P9/ML)
100
100
100
100
100
100
100
100
100
100
100
100
100
100
500
500
100
82901
Concentration
(P9/ML)
10
10
25
25
50
10
10
25
25
50
50
_ _
1 Provided as reference only; also see Tables 2 and 3 of Method 8290.
0023A - 25
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TABLE 2
SAMPLING QC PROCEDURES SUMMARY
QC Procedure
Sample equipment
calibrations
Dry gas meter sample
leak check
02 and C02 sampling
system leak check
AP meter leveling
Pitot tube leak check
Pitot tube orientation
check
Cyclonic flow check
Probe, filter, trap,
and silica gel impinger
are maintained at
specified temperature
ranges
Overall isokinetic
sampling rate
Sampling blanks
Frequency
See Sec. 6.3.1
Before and after
each test run
Once per test
Before and after
each test run
Before and after
each test run
Every test
Made at every
location
Every test
Every test
See Sec. 8.2
Criteria
See Sec. 6.3.1
0.00057 cmm (< 0.02
cfm) or 4% of sample
rate whichever is less
at highest vacuum
See Sees. 4.4 and 5.0
of EPA Method 3, or
equivalent for Method
3A
Level
No visible leak
observed at 75 mm (3
in.) H20 for 15 seconds
Pitot tube is level
with no visible
rotation from
perpendicular to flow
< 20° average offset
from perpendicular to
flow
See Sec. 4.0
± 10% of 100%
See Sec. 8.2
0023A - 26
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TABLE 3
REQUIREMENTS FOR ANALYTICAL PREPARATION,
SURROGATE RECOVERIES AND SAMPLE BLANKS
Item
Precleaning
filters
Precleaning
sorbent
Filter and
sorbent surrogate
spikes
Field blank
Method blank
Reagent blanks
Proof Blank
Description
Soxhlet extraction
Soxhlet extraction
Isotopically-labeled
compounds
Collect one for every
9 sample runs at each
test location
Prepared at analytical
laboratory (laboratory
blank). One per
analytical batch
One per lot of solvent
used. Archive for
possible analysis
One per set of
glassware. Archive
for possible analysis
(collect only if
requested by Agency)
Control Limit
Detection limits listed in
Sec. 9.2
Detection limits listed in
Sec. 9.2
70 to 130% recovery
< 5 times the Detection
limits
Criteria decided by
laboratory QA officer
Analyze only if requested
by Agency to determine
source of field blank
confirmation
Analyze only if requested
by Agency to determine
source of field blank
contamination
0023A - 27
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TABLE 4
SAMPLE EQUIPMENT CALIBRATION SUMMARY
Equipment
Primary WTM8 or
DGMa
Sample DGM
Sample DGM
Thermometers,
Thermocouples
Nozzle
Pitot tube
AP gauge (if not
an inclined
manometer)
Balance
Barometer
Procedure
Primary
calibration
Full calibration
Post calibration
Calibration
check
ID calibration
Wind tunnel
calibration or
construction
specifications
verification
See Sec. 2.2,
EPA Method 2
Calibration
check
Calibration
check
Frequency
Every 12
months
Every 6
months
After each
test program
Initially
Before every
test program
Before every
test program
Once/test
program
Initially
Initially
Control Limits
± 1% average
Y, < 2% from Yavg
Ypost < 5% from Yfull
± 2°C (3.6°F) at 3
point cal ibration
from reference
thermometer
Repeated
measurements
± 0.1 mm
(0.004 in.)
Specifications
listed in Sec. 4
of EPA Method 2
Within 5% of
reference at three
readings
Observed weight
< 0.5 g from Class
S weight
< 0.1 in. Hg from
primary barometer
aWTM = wet test meter; DGM = dry gas meter.
0023A - 28
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sac*
PCDD/PCDF Sampling Train Configuration
Figure 1
0023A - 29
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Flow
Direction
2 Piece
Configuration
28/12 Ball Joint
Water Jacket —
Condenser
28/12 Socket Joint
28/12 Ball Joint
Glass Wool Plug
Sorbent Trap
(20-40 gram Sorbent Capacity)
40 RC Glass
or Teflon® Frit
28/12 Socket Joint
1 Ptoc«
Configuration
Condenser and Sorbent Module Configurations
Figure 2
0023A - 30
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FIGURE 3
LU
O
i
o
(A
I
E
O
a
n
Q
O
i
a
o
a.
0023A - 31
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FIGURE 4
Container 2
(Front Half Rinse)
Container 1
(Filter)
I
Container 4
(Back Half Rinse)
Containers*
(Sorbent Trap)
Concentrate to
1-5mLat<37°C
Add Surrogate
Standards
Add Internal
Standards
Soxhlet Extract
f or18 Hours
Concentrate to
1-2mLat<37°C
Add Internal
Standards
Soxhlet Extract
for 18 Hours
Concentrate and
Bring to 5 mL
with Hexane
Concentrate and
Bring to 5 mL
with Hexane
r
Add Alternate
Standards
Archive Half in
Freezer for Possible
Repeat Analysis
~l
Perform Sample
Clean-up and
FracttonaDon
T
Add Recovery
Standards
Add Alternate
Standards
Archive Half in
Freezer for Possible
Repeat Analysis
Analyze with
06-5 Column
If TCOF is Found,
Confirm with 06-225
Column Analysis
Perform Sample
Clean-up and
Fractiortation
Add Recovery
Standards
T
Analyze with
06-5 Column
T
If TCOF is Found,
Confirm with 08-225
Column Analysis
Surrogate Standards are added to
the sorbent trap prior to sampling
PCDD/PCOF Analytical Summary Scheme
0023A - 32
Revision 1
January 1995
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APPENDIX A
RECOMMENDED AUDITING PROCEDURES
An audit is an independent assessment of data quality. Both performance
audits and system audits may be performed.
Performance Audit - A performance audit is conducted to evaluate
quantitatively the quality of data produced by the sampling, analysis, or
the total measurement system (sample collection, sample recovery, sample
analysis, and data processing).
Audit Sample - A performance audit sample contains tetra- through
octa-isomers of PCDD and PCDF. Audit samples are not normally
required.
Performance Audit of the Field Test - A field test performance audit
may be conducted by checking the dry gas meter for accuracy using
procedures located in the Quality Assurance Handbook for Air Pollution
Measurement Systems (EPA 600/4-77-027b). Performance audits on
thermocouple readings, AP gauges, barometric pressure gauges and
others, may also be conducted.
Performance Audit of Data Processing - The data processing procedures
may be audited by requiring the testing laboratory to provide an
example calculation for one sample run. This example calculation will
include all the calculations used to determine the emissions based on
the raw field and laboratory data.
System Audit - A system audit is an on-site, qualitative inspection and
review of the total measurement system.
The functions of the auditor are:
a) Observe procedures and techniques of the field team during sample
collection and sample recovery; and
b) Examine records of apparatus calibrations and other quality
control procedures used in sampling and analytical activities
When on-site, the auditor observes the source test team's overall
performance, including the following operations:
a) Setting the sampling system and leak checking the sample train and
pi tot tube;
b) Collecting the samples isokinetically;
c) Conducting the final leak checks; and
d) Sample documentation procedures, sample recovery, and preparation
of the samples for shipment.
0023A - 33 Revision 1
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METHOD 0031
SAMPLING METHOD FOR VOLATILE ORGANIC COMPOUNDS (SMVOC)
1.0 SCOPE AND APPLICATION
1.1 Method 0031 is used to determine volatile organic compounds in gaseous
emissions from a wide variety of stationary sources including hazardous waste
incinerators. The following compounds may be determined by this method:
Compound Boiling Point (°C) CAS No.'
Acrylonitrileb
Benzene
Bromodi chl oromethane
Carbon disulfide
Carbon tetrachloride
Chl orodi bromomethane
Chloroform
Chloroprene0
Di bromomethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans- 1 , 2-Di chl oroethene
1,2-Dichloropropane
1 , 3-Di chl oropropene
Methylene chloride
Tetrachl oroethene
Toluene
1,1,1-Trichloroethane
1 , 1 , 2-Tri chl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
8 Chemical Abstract Services
T r\ r> 1.1 <^ ^ SMA r>*\Tiil^^n*i+w ^»*/J vw
77
80
87
46
77
119-120 (? 748 mm Hg
61
59
97
57
83
32
48
96
106 @ 730 mm Hg
39
121
111
75
113
87
24
Registry Number.
p\*\s*+^lf44'V/ f\ £ 'V It 4 <•• /•• sxrvirt f\t I
107-13-1
71-43-2
75-27-4
75-15-0
56-23-5
124-48-1
67-66-3
126-99-8
74-95-3
75-34-3
107-06-2
75-35-4
156-60-5
78-87-5
542-75-6
75-09-2
127-18-4
108-88-3
71-55-6
79-00-5
79-01-6
75-69-4
some stationary sources.
c Reactive compound; may interact with the test matrix.
1.2 Method 0031 may be used to prepare volatile organic compounds that
have a boiling point between -15eC and 121°C. Field application for volatile
organic compounds with boiling points less than 0°C should be supported by data
obtained from laboratory gaseous dynamic spiking and gas chromatographic/mass
spectrometric (GC/MS) analysis according to Methods 5041 and 8260 to demonstrate
the efficiency of the sampling and analysis method.
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1.3 The method is not applicable to participates or aerosols since
isokinetic sampling is not performed. Isokinetic sampling is not required
because the volatile organic compounds are in the gas phase when they are
sampled. Dynamic spiking may require using reduced sample volumes collected at
flow rates between 250 and 500 mL/min, for a total sample volume of 20 L.
1.4 Application of Method 0031 is not restricted to those compounds in the
target analyte list, however, detection limits have been determined for these
compounds and acceptable method performance data have been obtained. Method 0031
may also be applied to the compounds listed in Table 1 if extra care is taken
because of the high volatility of these compounds.
1.5 Method 0031 is generally not applicable to polar water-soluble and
reactive volatile organic compounds. Examples of polar water-soluble and
reactive compounds are shown in Table 2. Other examples where Method 0030 (VOST)
sampling and analytical methodology has been used inappropriately include:
bromoform (boiling point 137°C, above the maximum limit allowed by the
methodology), ethylbenzene (136°C), 1,2,3-trichloropropane (156"C), xylenes
(~1408C), styrene (146eC), 1,1,2,2-tetrachloroethane (146°C at 746 mm Hg), and
the dichlorobenzenes (~ 175eC). Although successful analysis for these compounds
can be demonstrated by spiking sorbent tubes, the compounds will not be collected
quantitatively at the upper temperature limit for the operation of the SMVOC
train.
1.6 This method is applicable to the determination of volatile organic
compounds in the gaseous effluent of stationary sources such as hazardous waste
incinerators with an upper concentration limit per compound in the emissions of
approximately 1.5 parts per million (ppm). Method 0031 is not appropriate for
gaseous volatile organic compound concentrations above this limit, since
saturation of the analytical system or compound breakthrough in the field may
occur. Modifications of analytical methods to reduce the concentration of
compounds entering the gas chromatograph/mass spectrometer (GC/MS), such as
splitters or dilutions, may prevent saturation of the analytical system, but the
analytical data are not accurate if breakthrough has occurred during sampling.
The analysis of screening samples or distributive volume samples is recommended
to prevent analytical system saturation when high analyte concentrations may be
encountered.
1.7 The sensitivity of this method is dependent upon the level of
interferences in the sample matrix and the presence of detectable levels of
volatile organic compounds in the blanks. The target detection limit of this
method is 0.1 ng/m3 (ng/L) of gaseous effluent. The upper end of the range of
applicability of this method is limited by breakthrough of the volatile organic
compounds on the sorbent traps used to collect the sample and the ability of the
analytical system to respond within the linear range of the instrumentation.
Laboratory method development data have demonstrated a range of 0.1 to 100 jiig/m3
(ng/L) for selected volatile organic compounds collected on a set of sorbent
traps using a total sample volume of 20 L or less (see Sec. 2.3).
1.8 The SMVOC is designed to be operated at a sampling rate of 1 L/min
with traps being replaced every 20 min for a total sampling time of 2 hrs.
Analysis of the traps is carried out by thermal desorption purge-and-trap gas
chromatography/mass spectrometry (see Methods 5041 and 8260). Traps may be
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analyzed separately or combined onto one trap to improve detection limits.
Additional flow rates and sampling times are acceptable. For example, when less
than maximum detection ability is needed, it is acceptable to operate the SMVOC
at 0.5 L/min for a total of three 40-minute periods (two-hour total sampling
time). In this example, a two-hour sampling time is maintained, but the number
of sampling tubes which must be changed in the field is minimized, as is the
number of analyses which must be performed.
NOTE: The SMVOC sampling train may be operated no slower than 0.25 L/min, and
no faster than 1 L/min.
1.9 This method is restricted to use by, or under close supervision of,
trained analytical personnel experienced in sampling volatile organic compounds
in air. Each analyst must demonstrate the ability to generate acceptable results
with this method.
2.0 SUMMARY OF METHOD
2.1 This method employs a sampling module and meter box to withdraw a 20-L
sample of effluent gas containing volatile organic compounds from a stationary
source at a flow rate of 1 L/min, using a glass-lined probe heated to 130 + 5°C
and a sampling method for volatile organic compounds (SMVOC) train.
2.2 The gas stream is cooled to 20°C by passage through a water-cooled
condenser and volatile organic compounds are collected on a set of sorbent traps
(Tenax®-GC/Tenax®-GC/Anasorb®-747). Liquid condensate is collected in an
impinger placed between the two Tenax®-GC traps and the Anasorb®-747 trap. The
first and second traps contain 1.6 g of Tenax®-GC each and the third trap (back
trap) contains 5.0 g of Anasorb®-747. A total number of sorbent tube sets to
encompass a total sampling time of 2 hrs is collected: i.e., if a sampling rate
of 1 L/min for 20 minutes is used, a total of six sorbent tube sets will be
collected in 2 hr of sampling.
2.3 Alternative conditions for sample collection may be used, collecting
a sample volume of 20 L or less at a flow rate reduced from 1 L/min. (Operation
of the SMVOC under these conditions is referred to as SLO-SMVOC.) The SLO-SMVOC
may be used to collect 5 L of sample (0.25 mL/min for 20 min) or 20 L of sample
(0.5 L/min for 40 min) on each set of sorbent tubes. These smaller sample
volumes collected at lower flow rates should be considered when the boiling
points of the volatile organic compounds of interest are below O'C (see Table 1)
to prevent breakthrough. Refer to Sec. 2.2 for the total number of tube sets
collected per run.
3.0 INTERFERENCES
3.1 Interferences are encountered in the analytical methodology and arise
primarily from background contamination of sorbent traps prior to or after sample
collection. Other interferences may arise from exposure of the sorbent materials
to solvent vapors prior to assembly and exposure to significant concentrations
of volatile organic compounds in the ambient air at a stationary source site.
To avoid or minimize the low-level contamination of train components with
volatile organic compounds, care should be taken to avoid contact of all interior
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surfaces or train components with synthetic organic materials such as organic
solvents, and lubricating and sealing greases. Train components should be
carefully cleaned and conditioned according to the procedures described in this
protocol. The use of a sealed/enclosed sampling train is suggested but not
required (for example, a purged glove bag may be used). The use of blanks (Sec.
6.6) is essential to assess the extent of any contamination. Refer to Method
5041 for additional information on analytical interferences.
3.2 If the emission source has a high level of organic compounds in the
emissions matrix (for example, hydrocarbons present at levels of hundreds of
ppm), the presence of these volatile organic compounds may interfere with the
performance of the SMVOC analytical methodology. If the probability of
saturation of the analytical instrumentation exists, preliminary SMVOC screening
samples with distributive volumes may be necessary to help ensure that valid and
usable data will be obtained. To perform sampling according to distributive
volumes, samples of different volumes are collected (typically 5 L, 10 L, and 20
L) to verify that analyte concentrations are IX, 2X, and 4X. The concentrations
of artifacts produced by the sorbent will not vary with sample volume.
4.0 APPARATUS AND MATERIALS
4.1 Sampling train - A schematic of the principal components of the SMVOC
is shown in Figure 1. The SMVOC consists of a heated glass-lined probe, followed
by an isolation valve and charcoal trap, a water-cooled glass condenser, two
sorbent tubes containing Tenax®-GC (1.6 + 0.1 g each), an empty knock-out trap
for condensate removal, a second water-cooled glass condenser, a third sorbent
tube containing Anasorb®-747 (5.0 g ± 0.1 g), a silica gel drying tube, a
calibrated rotameter, a sampling pump, and a dry gas meter. The vacuum during
sampling and for leak-checking is monitored by pressure gauges which are in-line
with and downstream from the silica gel drying tube. The components of the
sampling train are described below.
4.1.1 Probe - The probe is made of stainless steel with a
borosilicate or quartz glass liner. The temperature of the probe is
maintained at 130'C ± 5'C or higher, but not so high that the sorbent
temperature exceeds 20"C. A water-cooled probe may be necessary at
elevated source temperatures to protect the probe and meet the required
sorbent temperature maximum. Isokinetic sample collection is not a
requirement for the use of SMVOC since the compounds of interest are in the
vapor phase at the point of sample collection. No nozzle is required, but
a plug of clean quartz wool (approximately 2.5 cm. (1 in.)) is inserted in
the probe to remove particulate matter.
NOTE: No stainless steel components should be in contact with the sample
stream.
4.1.2 Isolation valve - The isolation valve is a greaseless stopcock
(0.25 in. outer diameter stem is recommended) with a glass bore and sliding
Teflon® plug with Teflon® washers (Ace Glass 8193 or equivalent).
4.1.3 Condensers - The condensers (Ace Glass 5979-14 or equivalent)
must be of sufficient capacity to cool the gas stream to 20°C or less prior
to passage through the first sorbent tube. The top connection of the
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condenser must form a leak-free, vacuum-tight seal without using sealing
greases. Solverall® tube fittings and screw caps with Solverall® washers
(i in. OD, or equivalent) are recommended.
4.1.4 Sorbent tubes - See Figure 2 for a diagram of a SMVOC tube.
4.1.4.1 The first and second tubes of a three-tube set of
sorbent tubes should each be packed with 1.6 ± 0.1 g of Tenax®-GC
resin and the third tube of the set should be packed with 5.0 ± 0.1
g of Anasorb®-747. The tubes should be marked with an arrow to
indicate the direction of flow during sampling.
4.1.4.2 The sorbent tubes are glass tubes with approximate
dimensions of 10 cm x 1.6 cm ID. The tube is a single glass tube
which has the ends reduced in size to accommodate a i-in. Swagelok®
fitting. The sorbent is held in place by unsilanized clean glass wool
at each end of the sorbent layer. Threaded end caps are placed on the
sorbent tube after packing with sorbent to protect the sorbent from
contamination during storage and transport. In order to minimize tube
breakage, fittings are finger-tight plus an additional quarter of a
turn. Ceramic-filled Teflon® ferrules (Supeltex M2A or equivalent)
are used for tubes. Graphite ferrules (Supeltex M4 or equivalent) are
used if reconditioning of the tubes is necessary. The Swagelok® end
caps should be finger-tightened with the ferrules in place so that the
entire cap assembly may be turned as a unit. In order to seal the
assembly and avoid glass breakage, the cap assembly should be pushed
to the end of the glass and then backed off slightly before tightening
the cap with a wrench one quarter of a turn. Backing the cap assembly
off from the end of the tube will prevent chipping, cracking, or
breaking of the glass.
4.1.4.3 The sorbent tubes are placed in transport tubes
(capped culture tubes with glass wool and charcoal) for shipment. A
layer of clean charcoal is placed in the bottom of the transport tube
to absorb any volatile organics in the air in the transport tube. A
plug of cleaned glass wool (approximately 2.5 cm. (1 in.)) is placed
above the charcoal. The SMVOC tube, with both ends capped, is placed
in the transport tube, and a plug of cleaned glass wool (approximately
2.5 cm. (1 in.)) is placed on top of the SMVOC tube. The two glass
wool plugs cushion the SMVOC tube during shipping. The transport tube
is then sealed tightly with a Teflon®-!ined screw cap.
4.1.5 Metering system - The metering system for SMVOC consists of
a vacuum gauge, a pump, a calibrated rotameter for monitoring the sampling
flow rate, a dry gas meter (2% accuracy, with a minimum resolution of 0.01
L) at the required sampling rate, needle valves, and a temperature readout
device. Provisions should be made for monitoring the temperature of the
sample gas stream between the first condenser and the first sorbent tube,
since this temperature should not exceed 20°C. The temperature can be
monitored by placing a thermocouple on the exterior glass surface of the
outlet from the first condenser. The temperature at that point should be
less than 20°C. If the cooling is not sufficient, an alternative condenser
providing the necessary cooling capacity must be used.
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4.1.6 Sample transfer lines - All sample transfer lines connecting
the probe to the SMVOC shall be less than 1.52 m. (5 ft.) in length. The
sample transfer lines shall be heat-traced Teflon® or glass maintained at
130 ± 5°C. Connecting fittings must be capable of forming leak-free,
vacuum-tight connections without the use of sealing grease. All other
sample transfer lines used with the SMVOC shall be Teflon® with connecting
fittings that are capable of forming leak-free, vacuum-tight connections
without the use of sealing grease. These sample transfer lines should not
be reused at other emission sources.
4.2 Solverall® washers - All washers or gaskets used in SMVOC shall be
Teflon®-coated (Solverall® washers or equivalent; } in. stainless steel Swagelok®
fittings with Supeltex M2A ferrules may also be used). Prior to use, these
gaskets should be ultrasonically-cleaned with methanol and air-dried in a
contained/isolated organic vapor-free area. Gaskets should be stored in clean,
screw-capped containers prior to use.
4.3 Glass wool - Glass wool shall be Soxhlet-extracted for 8 to 16 hours
using methanol, and oven dried at 110°C before use. Glass wool should not be
silanized to prevent contamination of samples with siloxanes. Quartz wool is
recommended for high temperature applications.
4.4 Cold packs/ice - Ice or any commercially-available reusable liquids
or gels that can be frozen repeatedly are acceptable. These reusable liquids are
typically sold in plastic containers as "Blue Ice" or "Ice-Packs". Enough cold
packs or ice should be used to maintain tubes less than 10°C. If ice is used as
a coolant for the tubes, the tubes should be shielded from direct contact with
the ice so they will not become wet when the ice melts. Use of dry ice (solid
C02) for cooling tubes should be avoided; the sorbent tubes take up carbon
dioxide as the solid coolant vaporizes and the analytical system is vented when
the tubes are desorbed and analyzed. The tubes should not be stored at freezing
temperatures, since the seal between the glass and Teflon® fittings will be
compromised and diffusion of volatile organic compounds into the sorbent may
occur.
4.5 VOA vials - 40-mL glass vials with Teflon®-!ined screw caps are
required for recovery of condensate.
4.6 Teflon® squeeze bottles - Teflon® squeeze bottles should be washed
with a solution of a laboratory detergent, rinsed with hot tap water, then with
distilled water, then rinsed with clean purged water prior to use.
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. It is recommended that
blanks be taken of all reagents used in testing.
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5.2 2,6-Diphenyl-p-phenylene oxide polymer (Tenax®-GC, 35/60 mesh, or
equivalent).
5.2.1 New Tenax®-GC is Soxhlet-extracted for 24 hours with methanol.
The Tenax®-GC is dried for 6 hours in a vacuum oven at 50°C before use.
Thermal conditioning (Sec. 7.1.1) of the Tenax®-GC should be done prior to
blanking.
5.2.2 If reuse of Tenax®-GC is necessary, the polymer may be
extracted sequentially with methanol and pentane, dried in a vacuum oven,
and thermally reconditioned as described above. However, reused tubes must
meet the same criteria for cleanliness as new tubes. Reuse of sorbents is
not recommended. Common practice in laboratories where SMVOC tubes are
prepared commercially or where SMVOC sampling and analysis are done
extensively is not to reuse sorbents.
5.3 Anasorb®-747 - New Anasorb®-747 is used as it is received from the
manufacturer without preparation other than thermal conditioning pending a
Quality Control check (Sec. 7.1.1). Anasorb®-747 must not be reused. The
Anasorb®-747 should not be extracted with organic solvent prior to use as a
sorbent in the SMVOC.
5.4 Silica gel - Indicating type, 6-16 mesh. New silica gel may be used
as received from the vendor. Silica gel should not be reused for SMVOC.
5.5 Methanol, CH3OH - The methanol used for extracting the Tenax®-GC and
glass wool should be pesticide grade or equivalent.
5.6 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 SMVOC glassware cleaning - All glassware should be cleaned using the
following steps.
6.1.1 Sonicate in an ultrasonic bath for 1 hour in a solution of a
laboratory detergent such as Alconox®.
6.1.2 Rinse with copious amounts of hot tap water to remove all
detergent residue.
6.1.3 Rinse three times with HPLC grade water.
6.1.4 Oven dry at 110°C.
6.1.5 Cap for shipment using Teflon® tape or aluminum foil.
6.2 Assembly
The assembly and packing of the sorbent tubes should be carried out
in an area free of volatile organic material, such as a laboratory in which
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no organic solvents are handled or stored and in which the laboratory air
is charcoal filtered. Alternatively, an air-tight sealed glove box is
suggested.
6.3 Tenax®-GC tubes
6.3.1 The Tenax®-GC glass tubes and metal tube parts are cleaned and
stored (see Sec. 6.1). Ferrules are discarded but the metal unions are
cleaned by sonication in methanol. Tenax®-GC (1.6 ± 0.1 g) is weighed and
packed into each of the first two sorbent tubes which have unsilanized
cleaned glass wool in the downstream end. The Tenax®-GC is held in place
by inserting unsilanized cleaned glass wool. Each tube should be marked,
using an engraving tool, permanent marker or diamond-tipped pencil, with
an arrow to indicate the direction of sample flow during sampling, and a
serial number.
6.3.2 Conditioned sorbent tubes are capped and placed on cold packs
or ice for storage and transport. The temperature of the tubes during
storage and transport is maintained at a temperature of less than 10°C.
Conditioned tubes should be held for no more than 14 days before sampling,
to prevent the possibility of contamination.
6.4 Anasorb®-747 tubes - Anasorb®-747 (5.0 ± 0.1 g) is weighed and packed
into the third sorbent tube which also has unsilanized cleaned glass wool in the
downstream end. The Anasorb®-747 is held in place by inserting unsilanized
cleaned glass wool. Special care should be taken to conspicuously mark the
Anasorb®-747 tube with an arrow to indicate the direction of flow during
sampling, and a serial number.
6.5 Sample collection
6.5.1 After leak checking (see Sec. 6.5.3) but before the initiation
of sample collection, the probe shall be purged with stack gas. This purge
can be accomplished by attaching a pump to the isolation valve upstream of
the first condenser and drawing stack gas through the probe via the
isolation valve, so that the probe is purged of ambient air at the
initiation of sample collection.
6.5.2 Sample collection is accomplished by opening the valve at the
inlet to the first condenser (see Figure 1), turning on the pump, and
sampling at a rate of 1 L/min (or slower rate, if desired, according to the
guidelines for SLO-SMVOC) for 20 minutes (or an appropriately longer
period, if slower sampling rates are used). The volume of sample for any
set of traps should not exceed 20 liters. The end caps of the sorbent
tubes should be placed in a clean screw-capped glass container during
sample collection to prevent contamination.
6.5.3 Following completion of sample collection, the SMVOC is leak-
checked a second time at the highest vacuum encountered during the sampling
run to minimize the chance of vacuum desorption of volatile organic
compounds from Tenax®-GC. The sample is considered invalid if the leak
test does not meet specifications. The train is returned to atmospheric
pressure and the set of sorbent tubes is removed. The end caps are
replaced and the tubes are placed in an organic-free environment and
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maintained at a temperature less than 10°C for storage and transport. The
set of tubes and any condensate collected (see Sec. 6.5.4) are placed in
self-sealing plastic storage bags.
In the laboratory, tubes are maintained in a clean, organic vapor-free
environment at a temperature less than 10°C until analysis. The maximum
storage time between sampling and analysis of the tubes should be 14 days.
The rate of loss of sorbed volatile organic compounds from the tubes is
both compound-specific and source-specific. A 14-day period is chosen for
the holding time before analysis to provide a reasonably conservative
guideline for quantitative analysis of the volatile organic compounds which
have been sampled.
NOTE: To prevent breakage and/or loosening of the seals at the end of the
tubes, SMVOC tubes should not be stored in a freezer or over dry
ice. A solvent-free refrigerator (no cooler than 4°C) is
appropriate for storage of the tubes until analysis.
6.5.4 The condensate is recovered by transferring any liquid
contained in the knock-out trap to a 40-mL VOA vial and rinsing the
knock-out trap three times with a minimum volume of organic-free reagent
water (Sec. 5.6) and adding the rinses to the VOA vial. If necessary,
water should be added to eliminate headspace in the vial. If there is
sufficient condensate to fill more than one vial, two vials should be used.
The VOA vials containing the condensate are placed, with the set of tubes,
in a self-sealing plastic storage bag and maintained at a temperature less
than 10°C for storage and transport until analysis. The condensate is
analyzed by Method 8260. Refer to Method 8260 for details on analytical
procedures.
6.5.5 A new set of tubes is placed in the SMVOC, the SMVOC is
leak-checked, and the sample collection process repeated as described
above. Sample collection continues until sufficient samples to encompass
a two-hour sampling period have been collected. If samples are taken at
a sampling rate of 1 L/minute, a two hour sampling period will result in
the collection of six sets of tubes. If SLO-SMVOC procedures are used,
fewer than six sets of tubes will be sampled over a two-hour period.
6.6 Blanks
6.6.1 Field blanks - Blank Tenax®-GC and Anasorb®-747 tubes are
attached to the sampling train while the train is leak-checked. The tubes
are removed and stored with the sample tubes. At least one field blank
should be collected for every two-hour sampling period.
6.6.2 Trip blanks - At least one set of blank tubes (two Tenax®-GC,
one Anasorb®-747) should be included with each shipment of tubes to a
stationary source sampling site. These trip blanks should be treated like
any other tubes except that the end caps will not be removed during storage
at the site. This set of tubes should be analyzed to assess contamination
which may occur during storage and shipment.
6.6.3 Laboratory blanks - One set of blank tubes (two Tenax®-GC, one
Anasorb®-747) should remain in the laboratory using the method of storage
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which is used for field samples. ihese laboratory blanks should be from
the same batch of sorbent as used for the field blanks, trip blanks and
collected samples. If the field and trip blanks contain high
concentrations of contaminants (e.g., greater than 2 ng of a particular
volatile organic compound], the laboratory blank should be analyzed in
order to identify the source of contamination.
7.0 PROCEDURE
7.1 Tube condition!ng
7.1.1 In a desorption oven, the sorbent tubes are connected to a
source of organic-free nitrogen. Nitrogen is passed through each tube at
a flow rate of 80-100 mL/min while the tubes are heated. Anasorb®-747 is
thermally conditioned for 18-24 hours at 300°C, under a nitrogen flow rate
of 80-100 mL/min. Tenax®-GC is thermally conditioned at 220°C for 8-12
hours at a nitrogen flow rate of 80-100 mL/min. The actual length of time
required for the conditioning period may be determined based on the
adequacy of the resulting blank checks of the conditioned tubes. Method
5041 (modified to use a sorbent desorption temperature of 250°C) and Method
8260 may be used to perform a blank check of each set of sampling tubes to
ensure cleanliness.
7.1.2 An acceptable blank level is less than or equal to (<} Method
Detection Limits for Method 5041/8260 (see Method 8260 for Method Detection
Limits). A general guideline of analyte values less than 2 ng for any
volatile organic compound may be used as a criterion of cleanliness.
7.1.3 After conditioning, tubes are sealed and placed on cold packs
or ice (maintained at a temperature less than 10°C) until sampling is
completed. Conditioned tubes should be held for no more than 14 days
before sampling, to prevent the possibility of contamination.
7.2 Pretest preparation
7.2.1 All train components should be cleaned and assembled as
previously described. A dry gas meter should be calibrated within 30 days
prior to use, using a standard orifice, or other approved calibration
device/meter.
7.2.2 The SMVOC is assembled according to the schematic diagram in
Figure 1. Cooling water should be circulated to the condensers and the
temperature of the cooling water must be iow enough to maintain the
temperature of the gas entering the sorbent below 20°C.
7.3 Leak-checking
7.3.1 To leak-check the entire train, it is necessary to leak-check
from the probe to the pump. In order to adequately represent actual
sampling conditions, a leak-check should be performed with the pump on and
the leak rate measured in liters per minute (Lpm) on the dry gas meter.
After the desired vacuum is reached, the pump is isolated from the train
to check for leaks.
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7.3.2 Ensure that all connections are tight and that the train is
assembled correctly with sorbent cartridges properly assembled and in the
right direction for sampling. Seal the end of the probe and turn the
isolation valve to the sample/open position. Turn on the pump and adjust
the vacuum to 25.4 cm above normal operating pressure (38 cm Hg should be
sufficient as 12.7 cm or less is normal). Prior to leak-checking, verify
that the fine adjust valve on the meter box is partially opened and that
the coarse adjust valve is almost closed to prevent backflushing of any
condensate during final leak checks as the valves will need to be adjusted
to increase vacuum rather than decrease vacuum. Allow the rotameter on the
meter box to drop to zero and the pressure on the water column gauge
(represents the pressure inside the dry gas meter) to stabilize. The pump
is isolated by shutting off the needle valve. Record the leak rate
directly from the dry gas meter and time for one minute using a stopwatch.
The leak rate must be less than 0.02 Lpm. This value should be sufficient
as it is less than 1% of the sample rate.
7.3.3 Upon completion of the leak check, turn off the pump and
release the pressure/vacuum in the train by turning the isolation valve to
the purge position and allowing ambient air (filtered with charcoal or
equivalent) to enter the train. The initial leak-check should be above
normal operating pressure. The final leak-check (following collection of
20 L of sample) should be at least at the highest vacuum encountered during
the run.
NOTE: The volume of air pulled through the SMVOC during leak-checking
procedures prior to sampling should be less than 2.5% of the total
volume sampled. If a volume greater than 2.5% of the total
sampling volume is pulled through the SMVOC in obtaining a
successful leak check, the sorbent tubes used during this leak
check must be discarded and a successful leak check with a minimum
volume of gas pulled through the train must be obtained with a new
set of sampling tubes in place.
7.4 Sample collection - Sample collection procedures are described in Sec.
6.5.
7.5 Analytical procedure - Samples are analyzed by Methods 5041 and 8260.
In these methods, adapted for a three-tube SMVOC, the sorbent tubes are spiked
with surrogates, internal standards are spiked into the purge water, and the
tube(s) thermally desorbed at 250°C under a purge of organic-free helium. The
tubes may all be analyzed individually, or the Tenax® tubes may be analyzed as
a pair with the Anasorb® tube analyzed separately. The gaseous effluent from the
tubes is bubbled through purged organic-free reagent water (Sec. 5.6) and trapped
on an analytical sorbent trap in a purge-and-trap unit. After desorption, the
analytical sorbent trap is heated rapidly and the gas flow from the analytical
trap is directed to the head of a wide-bore capillary column (Method 5041) under
subambient conditions. The volatile organic compounds desorbed from the
analytical trap are separated by temperature-programmed gas chromatography and
detected by continuously-scanning low resolution mass spectrometry (Method 8260).
Concentrations of volatile organic compounds are calculated from a multipoint
calibration curve, using the method of response factors. Refer to Method 8260
for details.
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7.6 Calculations
7.6.1 The following nomenclature is used in the calculation of
sample volume:
Pbar = Barometric pressure at the exit orifice of the dry gas meter,
mm (in.) Hg.
Pstd = Standard absolute pressure, 760 mm Hg.
Tm = Dry gas meter average absolute temperature, °K (°R)
Tstd = Standard absolute temperature, 293°K (528°R)
Vm = Dry gas volume measured by dry gas meter, dcm (dcf)
V
mi.idi
= Dry gas volume measured by dry gas meter, corrected to
standard conditions, dscm (dscf)
y = Dry gas meter calibration factor
7.6.2 The volume of gas sampled is calculated as follows:
T P V P
u _ ii std bar _ i/ m bar
m.,rf nn ' T r\ 1 ' T
m std
where:
KT = 0.3858°K/mm Hg for metric units or
K, = 17.64°R/in. Hg for English units.
7.6.3 The concentration of volatile organic compound (CPD) in the
stack sample (Cg) is calculated as follows:
Total weight of CPD in sample (i.e., analytical measurement from VOST tubes)
\j — — -- ~~" ••— - - .,— .
9 Volume of sample at standard conditions, dscm
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8.0 QUALITY CONTROL
8.1 Prior to actual sampling on-site, all of the applicable sampling
equipment should be thoroughly checked to ensure that each component is clean and
operable. Each of the equipment calibration data forms should be reviewed for
completeness and adequacy to ensure the acceptability of the equipment. Each
component of the sampling system should be carefully packed for shipment. Upon
arrival on-site, the equipment should be unloaded, inspected for possible damage,
and then assembled for use.
8.2 The following quality control (QC) checks are applicable to the
sampling procedures:
8.2.1 Each sampling train must be visually inspected for proper
assembly before every use.
8,2.2 All sampling data should be recorded on standard data forms
which may serve as a pretest checklist.
8.2.3 The temperature measurement system should be visually checked
for damage and operability by measuring the ambient temperature.
8.2.4 All sampling data and calculations should be recorded on
Preformatted data sheets.
8.2.5 All glassware for SMVOC should be cleaned according to the
procedure in Sec. 6.1.
8.2.6 Ten percent of the SMVOC tubes should be subjected to GC/MS
QC measurements. No analytes should be detected at concentrations above
method detection limits in unused SMVOC tubes. If these quality control
tests are performed by the manufacturer, documentation should be obtained
from the commercial supplier and retained.
8.2.7 All cleaned glassware, hardware, and prepared sorbent traps
should be kept closed with ground-glass caps or Teflon® tape until assembly
of the sampling train in the field. The sorbent traps should be recapped
immediately after each set of samples is collected.
8.2.8 Prior to sampling, the Tenax®-GC and Anasorb®-747 tubes should
be spiked with the compounds of interest to ensure that they can be
thermally desorbed under laboratory conditions. This spiking is necessary
but not sufficient. The compound must still be sampled from the source.
8.2.9 Assembly and recovery of the sampling trains must be performed
in an environment as free from uncontrolled dust and solvent vapors as
possible.
8.2.10 Blanks (field, trip, laboratory) must be collected.
8.2.11 The entire sampling train should be leak-checked before and
after each run. If the sampling train is moved from one sampling port to
another during a run, the train should be leak-checked before and after the
move.
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8.2.12 Dry gas meter readings, temperature readings, and pump vacuum
readings should be made during sampling and recorded in intervals no
greater than 5 minutes.
8.2.13 Sorbent traps should be used for sampling within two weeks of
preparation.
8.2.14 During sample collection, the gas stream temperature at the
inlet to the first sorbent trap must be maintained at or below 20°C.
8.2.15 All sample traps should be stored under refrigeration or on
ice or cold packs (temperature maintained less than 10°C) until ready for
analysis.
8.3 QC for analytical procedures
8.3.1 Calibration standards should be prepared at five concentration
levels for each analyte of interest. Compounds of interest, surrogate
compounds, and internal standards are spiked into the purge water for
generation of a multipoint calibration curve. When samples are analyzed,
surrogate compounds are spiked onto the sampling tubes using flash
vaporization techniques (Method 5041), but internal standards are spiked
into the purge water. Response factors for each compound are calculated
and these response factors are used for the calculation of analytical
results. Refer to Methods 5041 and 8260 for detailed analytical QC
procedures for analysis of samples.
8.3.2 To establish the precision and accuracy of the analysis,
triplicate paired Tenax®-GC tubes should be spiked with analytical
surrogate volatile organic compounds using flash evaporation and analyzed
immediately following the initial calibration and before sample analysis.
The spiking level should be at the expected level of volatile organic
compounds in the stationary source. The spiking standard must be prepared
from stock standards separate from those used for calibration. Recovery
for each volatile organic compound and surrogate should be within 50% to
150% of spiked value. The relative standard deviation associated with each
analyte should be less than 25 percent.
8.3.3 The average recovery from the initial precision and accuracy
determinations should be used as an acceptance criterion for sample
results. The surrogate recovery in each sample should be within three
standard deviations of the average recovery obtained from the initial
precision and accuracy determinations.
8.3.4 An EPA performance audit should be completed during a trial
burn as a check on the entire SMVOC system. The audit results should agree
within 50% to 150% of the expected value for each specific compound of
interest. This audit consists of collecting a gas sample containing one
or more volatile organic compounds in the SMVOC from an EPA audit gas
cylinder. Collection of the audit sample in the SMVOC may be conducted
either in the laboratory or at the field test site. Analysis of the SMVOC
audit sample must be by the same person, at the same time, and with the
same analytical procedure as used for the regular SMVOC samples from the
field test.
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9.0 METHOD PERFORMANCE
See Method 8260.
10.0 REFERENCES
1. "Field Test of a Generic Method for Halogenated Hydrocarbons" (U.S.
Environmental Protection Agency). EPA 600/R93/101, June 1993.
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TABLE 1
HIGH VOLATILITY ORGANIC COMPOUNDS3
Compound Boiling Point
Bromomethane 4°C
Chloroethane 12°C
Vinyl bromide 16°C, at 750 mm
Vinyl chloride -13.4°C
aUse of SLO-SMVOC may be helpful
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TABLE 2
COMPOUNDS FOR WHICH METHOD 0031 IS NOT APPLICABLE
Compounds
Boiling Point
Allyl chloride
Acetone
Methyl ethyl ketone
Chloromethane
Epichlorohydrin
45°C
56°C
80°C
-24°C
116°C
Chloromethyl methyl ether 56°C
bis(Chloromethyl) ether 106°C
Acetonitrile 82°C
Acetaldehyde 21°C
Acrolein 53°C
Methanol 65°C
Ethanol 78°C
Isopropyl alcohol 82°C
Comment
Reactive compounds; interacts
with test matrix to yield poor
recoveries and poor
reproducibility
Polar, water soluble
Polar, water soluble
Reactive compounds; interacts
with test matrix to yield poor
recoveries and poor
reproducibility
Not amenable to SMVOC
analytical procedure
Not amenable to SMVOC
analytical procedure
Not amenable to SMVOC
analytical procedure
Polar, water soluble
Polar, water soluble, reactive
Polar, water soluble, reactive
Polar, water soluble
Polar, water soluble
Polar, water soluble
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FIGURE 1
SCHEMATIC OF SAMPLING METHOD FOR VOLATILE ORGANIC COMPOUNDS (SMVOC) TRAIN
Glass
Wool
Filter
Stack
Healed Glass
Lined Probe
Three Way 1/8'OD
GlasaATeflon Teflon Line
Valve A
Condensers
Condensale
Trap
Condenser
Ice Balh
Temperature
Indicators
f f
Exhaust
Meter Box
Sampling Module
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FIGURE 2
SMVOC TUBE
Glass
Wool
0.6 cm 00
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METHOD 0040
SAMPLING OF PRINCIPAL ORGANIC HAZARDOUS CONSTITUENTS
FROM COMBUSTION SOURCES USING TEDLAR® BAGS
1.0 SCOPE AND APPLICATION
1.1 This method establishes standardized test conditions and sample
handling procedures for the collection of volatile organic compounds collected
from effluent gas samples from stationary sources, such as hazardous waste
incinerators and other combustion sources, using time-integrated evacuated
Tedlar® bags. The compounds listed below can be collected by this method. This
is a sample collection method and does not directly address the analysis of these
samples. Gas chromatography/mass spectrometry (GC/MS) (Method 8260) is the
recommended analytical technique because of its ability to provide positive
identification of compounds in complex mixtures such as stack gas.
Compound CAS Registry No.
Dichlorodifluoromethane 75-71-8
Vinyl chloride 75-01-4
1,3-Butadiene 106-99-0
l,2-Dichloro-l,l,2,2-tetrafluoroethane 76-14-2
Methyl bromide 74-83-9
Trichlorofluoromethane 353-54-8
1,1-Dichloroethene 75-35-4
Methylene chloride 75-09-2
1,1,2-Trichloro-trifluoroethane 76-13-1
Chloroform 67-66-3
1,1,1-Trichloroethane 71-55-6
Carbon tetrachloride 56-23-5
Benzene 71-43-2
Trichloroethene 79-01-6
1,2-Dichloropropane 78-87-5
Toluene 108-88-3
Tetrachloroethene 127-18-4
1.2 This method is not applicable to the collection of samples in areas
where there is an explosion hazard. Substitution of intrinsically safe equipment
or procedures for the equipment or procedures described in this method will not
be sufficient to adapt this method for use in areas where there is an explosion
hazard. Additional modifications to the sampling and analytical protocols may
be required.
1.3 This method does not employ isokinetic sampling and therefore is not
applicable to the collection of highly water soluble volatile organic compounds
contained in an aerosol of water. This method uses either constant or
proportional rate sampling, depending upon the extent of the variability of the
emission flow rate.
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1.4 This method is restricted to use by, or under the close supervision
of, trained analytical personnel experienced in sampling organic compounds in
air. Each analyst must demonstrate the ability to generate acceptable results
with this method.
1.5 Each compound for which this method can be considered shall meet the
criteria listed in Sees. 1.5.1 - 1.5.3, below. Method 8260 provides boiling
points, condensation points (calculated from vapor pressure) at 20°C, and
estimated instrument detection limits. This method is not limited to the
compounds in the target analyte list, however, stability and recovery shall be
demonstrated when compounds other than those listed in Sec. 1.1 are to be
sampled.
1.5.1 The compound must have a boiling point < 121°C.
1.5.2 The compound must have a concentration in the stack gas below
the condensation point.
1.5.3 During validation studies, the loss of the compound from a
Tedlar® bag must be less than 20% over a 72-hour storage time.
2.0 SUMMARY OF METHOD
2.1 A representative sample is drawn from a source through a heated sample
probe and filter.
2.2 The sample then passes through a heated 3-way valve and into a
condenser where the moisture and condensable components are removed from the gas
stream and collected in a trap.
2.3 The sample is collected in a Tedlar® bag held in a rigid, opaque
container.
2.4 The dry gas sample and the corresponding condensate are then
transported together to a GC/MS. A mass spectrometer is most suited for the
analysis and quantitation of complex mixtures of volatile organic compounds. The
total amount of the analyte in the sample is determined by summing the individual
amounts in the bag and the condensate. A flow chart of the procedure is given
at the end of this method.
3.0 INTERFERENCES
3.1 The materials from which the Tedlar® bag is constructed may contribute
background hydrocarbon contamination. Purging the bag with air or N2 may reduce
the concentration of these hydrocarbons. Exposure of the bag to direct sunlight
may increase the concentration of these hydrocarbons. Therefore, the bag must
be protected from exposure to sunlight by using an opaque container to house the
bag during sampling and shipping.
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3.2 Components of the source emissions other than the target compounds may
interfere. Interferants may be differentiated from the target compounds during
mass spectrometric analysis.
3.3 Common problems that can invalidate Tedlar® bag sampling data and
techniques to remedy these problems are listed in Table 1.
3.4 Available stability data suggest that this method may not perform well
in sampling streams containing polar and reactive compounds like methyl ethyl
ketone, formaldehyde, methanol, 1-butene, and acetone. The use of this method
to sample these compounds needs to be evaluated before sampling.
4.0 APPARATUS AND MATERIALS
4.1 Tedlar® bag sampling train - A detailed schematic of the principal
components of the sampling train is shown in Figure 2.
4.1.1 The sampling train (Figure 2) consists of a glass-lined probe,
a heated glass or Teflon® filter holder and quartz filter attached to one
of two inlets of a glass and Teflon® 3-way isolation valve (Figures 3 and
4). The second valve inlet is used to release system pressure after leak
checks. This valve is connected to a charcoal trap, which filters incoming
air. The outlet of the isolation valve is connected to a glass, water-
cooled, coil-type condenser and a glass condensate trap for removal and
collection of condensable liquids present in the gas stream. A 1/4-in. OD
x 1/8-in. ID Teflon® transfer line connects the condensate trap to a second
3-way isolation valve and the isolation valve to a Tedlar® bag contained
in a rigid, air-tight container for sampling, storage, and shipping. The
bag container is connected to a control console with 1/4-in. OD x 1/8-in.
ID vacuum line by means of 1/4-in. Teflon® connectors at each end. A
charcoal trap is placed in the vacuum line between the bag container and
the control console to protect the console and sampling personnel from
hazardous emissions in case of bag rupture during sampling.
4.1.2 The vacuum required to operate this system is provided by a
leak-free diaphragm pump contained in the control console (Figure 5). When
the pump is turned on, the space between the inner walls of the bag
container and the Tedlar® bag is evacuated, placing the system under
negative pressure, which pulls the sample through the sampling train and
into the Tedlar® bag. The sampling train vacuum is monitored with a vacuum
gauge installed in-line between the vacuum line and the coarse adjustment
valve mounted in the control console.
4.1.3 Sample flow rate is regulated by adjusting the coarse and fine
valves on the control console. The coarse adjustment valve controls the
sample inlet volume and rate and isolates the vacuum line, vacuum gauge,
and sample train from the pump and other console components during leak
checks. Sample volume is measured with a calibrated dry gas meter
contained in the control console. Sampling rate is monitored by a
rotometer, contained in the control console, and is installed on the outlet
side of the dry gas meter.
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4.1.4 The source, probe, filter, and condenser temperatures are
monitored by Type J or K thermocouples using the temperature readout in the
control console. Probe heater temperature is regulated by the temperature
controller provided in the control console (Figure 5).
4.1.5 The velocity pressure and temperature of the source gases are
measured using a standard or S-type pitot tube connected to a manometer
with 1/4-in. OD x 1/8-in. ID tubing, in accordance with EPA Method 2. The
source velocity pressure and temperature must be monitored during sampling
and the sampling rate adjusted proportionally to changes in the flue gas
velocity (Sec. 7.5.1.2).
4.2 Sample train components
4.2.1 Probe assembly - The probe assembly consists of a length of
heated and insulated borosilicate glass tube inside a length of stainless
steel tubing. The probe temperature shall be maintained between 130°C and
140*C in order to prevent damage to Teflon® lines and to facilitate
efficient cooling of the gases in the condenser. The stainless steel
sheath must be cooled with water when the source temperature approaches or
exceeds 140°C.
4.2.2 Particulate filter - Particulate matter from the sample gas
stream exiting the probe is collected on a quartz filter substrate in a
heated 47-mm Teflon® or glass filter holder. Use clean filters in order
to prevent sample contamination. The particulate matter itself is not
analyzed or archived. However, removal of particulate matter provides a
cleaner sample for analysis. All connections between the probe and
particulate filter shall be heated to maintain the temperature between
130'C and 140°C so that compounds remain in the volatile phase. Heat-
wrapped Teflon® unions with stainless steel nuts and Teflon® ferrules are
recommended for all heated connections.
4.2.3 Isolation valves - A typical isolation valve is shown in
Figure 3. The isolation valves shall be constructed of Teflon® or glass
with Teflon® stopcocks to provide gas-tight seals without the use of
sealing greases. The probe and bag isolation valves are of identical
design and materials and are therefore interchangeable. The probe
isolation valve provides for the attachment of a charcoal or similar purge
trap to allow filtered ambient air to enter the train when returning the
train to ambient pressure after leak checks. This valve directly connects
the probe and filter assembly to the condenser inlet and must be heated to
between 130"C and 140°C. The bag isolation valve allows the bag to be
opened for sampling or evacuation and isolated and sealed for leak checks
or system purges.
4.2.4 Condenser - Use a jacketed, water-cooled, coil-type glass
condenser with a volume of at least 125 ml. The condenser shall have
sufficient capacity to maintain the temperature of the sample gas stream
between 20'C and 4*C to ensure proper removal and collection of condensable
moisture in the effluent gas sample. The cooled sample gas stream
temperature should not exceed ambient temperature. All condenser
connections must form a leak-free, vacuum-tight seal without using sealing
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greases. Stainless steel fittings are not permitted, and Teflon® unions
or washers with screw caps are recommended.
4.2.5 Condensate trap - A glass Erlenmeyer distilling flask with
threaded screw cap connections, Teflon® seals, and a minimum volume of
125 ml may be used to trap condensate. All connections on the condenser
and trap shall be sized to accept 1/4-in. OD x 1/8-in. ID Teflon® or glass
fittings. The stem from the condenser must be positioned to within 0.5-in
from the bottom of the condensate trap.
4.2.6 Sample transfer lines and connection fittings - All sample
transfer lines connecting components shall be less than 5 ft long and
constructed of 1/4-in. OD x 1/8-in. ID Teflon® tubing or glass. All sample
lines upstream of the condenser and condensate trap must be heated and the
temperature maintained between 130°C and 140°C. Use Teflon® fittings for
connections between various train components to provide leak-free, vacuum-
tight connections without the use of sealing grease. New tubing should be
used for each separate test series or condition to prevent cross
contamination of sample compounds.
4.2.7 Tedlar® storage bag - Choose a bag size according to the
guidelines provided in Sec. 7.2.4. In order to minimize wall effects, the
sample volume must fill at least 80% of the bag capacity. The recommended
size range for bags is 25 L to 35 L. Small bags (< 25 L) are easier to
store and transport but may have insufficient volume for proportional
sampling. In addition, accurate volumetric measurement is difficult with
smaller bags. Large bags (> 50 L) lack portability, but may be required
under certain conditions, such as during proportional sampling and for
sampling sources requiring high sample rates.
4.2.8 Evacuated container (bag container) - Use any rigid, air-tight
metal or plastic (e.g., PVC®/Polyethylene®/Nalgene®) drums or glass
containers to house the Tedlar® bag during sampling, storage, and
transport. The container must be constructed so that it can easily be
assembled and disassembled (for bag removal). The container must be able
to hold a negative pressure of at least 10 in. H20. The bag container must
be at least 20% smaller than the Tedlar® bag being used but must be large
enough to hold the volume of sample required (e.g., for a sample size of
20 L, a 25-L Tedlar® bag inside a 20-L container provides sufficient volume
without danger of overinflating the bag).
Containers must not have staples, sharp edges, or metal closures which
might damage bags. The container should also be constructed of a material
that shields the sample from exposure to sunlight to protect the bag and
its contents from ultra-violet light. A viewing port or other means of
observing the flexible bag during sampling is desirable. During storage
and transport, the viewing port shall be covered with opaque material.
4.2.9 Vacuum lines - Use Tygon®, Poly®, Nylon®, or similar tubing
capable of maintaining at least 10-in. H20 negative pressure without
collapse as vacuum lines. Tubing should be 1/4-in. OD x 1/8-in. ID size
to minimize volume and ensure compatibility of connection fittings
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throughout the train. Stainless steel fittings and valves may be used for
vacuum line connections but may not be used in the sampling line.
4.2.10 Control console (meter system) - The metering system required
for this method is readily available in the form of the control
console/meter box (e.g., Nutech Model 280.01B) from a Volatile Organic
Sampling Train (VOST, Method 0030), and shall consist of the components
pictured in Figure 5.
4.2.10.1 Vacuum gauge (meter pressure) - Use a direct
reading, mechanical vacuum gauge capable of measuring pressures of at
least 15 in. Hg with 1-in. or smaller increments to monitor system
vacuum during sampling and leak checking the bag, the container, and
the sampling train.
4.2.10.2 Sample flow rate adjustment valves - Coarse and
fine adjustment valves are provided. The coarse adjustment valve
controls volume and rate of sample flow and isolates the control
console from the sampling train and vacuum line during leak checks.
The fine adjustment valve controls sample rate and system vacuum.
Closing the valve (clockwise) increases train vacuum and sample flow
rate. Opening the valve (counterclockwise) decreases train vacuum and
sample flow rate.
4.2.10.3 Pump - Use a leak-free diaphragm pump or equivalent
that is capable of pulling and maintaining a vacuum of at least 15 in.
Hg and a flow rate of at least 1 liter per minute (Lpm).
4.2.10.4 Calibrated dry gas meter - The control console
contains a calibrated dry gas meter (Singer Model 802/American Meter
Model 602 or equivalent) capable of reading 1 L per revolution with
0.1-L increments, and provides accurate measurement of the volume of
the sample collected.
4.2.10.5 Flow meter - Use a rotometer with a glass tube and
a glass, Teflon®, or sapphire float ball of suitable range (0-5 Lpm)
to measure the sample flow rate. The flow meter shall be accurate to
within 5% over the selected range. A range of ± 25% of the desired
sampling rate is suggested to ensure greater accuracy of readings and
a better range for adjustment of the sampling rate (proportional to
the source gas stream velocity). The rotometer is installed at the
outlet of the dry gas meter in the console.
4.2.10.6 Thermocouples and temperature read-out device - Use
a sufficient number and length of type J or K thermocouples. The 10-
channel (1 to 4 remote; 5 dry gas meter, 6 to 10 spares) digital
thermocouple read-out provided in the control console displays the
source, probe, filter, and condenser temperatures.
4.2.10.7 Heat controller - Use a rheostat or digital
temperature controller (e.g., Fuji PYZ4 or equivalent) to regulate
probe heat temperatures.
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4.2.11 Pitot tube probe - A standard or S-type pitot tube must be
used for pretest and post-test velocity traverses and to monitor flow so
that the sampling rate can be regulated proportionally to the source gas
velocity throughout the length of the sampling run.
4.2.12 Pressure gauge (manometer) - Use a water- or oil-filled U-tube
or inclined manometer capable of measuring to at least 10 in. H20 and
accurate to within 0.1 in. H20 for monitoring and measuring the source gas
velocity.
4.2.13 Barometer - Use an aneroid or other barometer capable of
measuring atmospheric pressure to within 0.1 in. Hg of actual barometric
pressure.
4.2.14 Charcoal absorbent traps - Use charcoal traps to absorb
organic compounds in the atmosphere at the site. One charcoal trap is
attached to the probe isolation valve and filters incoming air when
releasing vacuum to prevent contamination of the train during leak checks.
A second charcoal trap is located in the vacuum line and filters any gas
exiting the sample train to protect sampling personnel in case of bag
rupture. Any readily available, ready made charcoal tube similar to a VOST
tube may be used.
4.2.15 Stopwatch - Use any stopwatch capable of measuring 1 second,
to time sample collection.
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 Water - Water used for sample train preparation shall be distilled and
deionized. Water used for rinses during recovery of condensate shall be
prepurged high performance liquid chromatography (HPLC)-grade. Clean, clear tap
water may be used as condenser cooling water.
5.3 Nitric acid, HN03 (10%) - reagent grade.
5.4 Charcoal - SKC petroleum-base charcoal, or equivalent. A mesh size
of 6-14 is recommended. New or reused charcoal may be used for each run series
or test condition. Reused charcoal must be reconditioned using the same criteria
specified in VOST (Method 0030).
5.5 Methanol - Spectrometric-grade, or equivalent.
5.6 Field spiking standards - Appropriate gas cylinders containing the
target components of interest in known concentrations (highest purity available)
for field spiking.
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6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Pretest preparation
6.1.1 Glassware - Before sampling, prepare the glass components of
the train by cleaning with non-ionic detergent (e.g., Alconox) and hot
water in an ultrasonic bath. Rinse each component three times with
distilled, deionized water, then rinse three times with 10% HN03, followed
by an additional three rinses with distilled, deionized water. Dry in an
oven at 130°C for 2 hours.
6.1.2 Sample lines and bag containers - Treat all Teflon® lines,
fittings, and the sample bag containers as outlined in Sees. 6.1.1, but air
dry these components in an area free of organic compounds rather than in
an oven. Use clean Teflon® tubing for each test series or condition. Hand
wash the bag containers.
6.1.3 Bag cleaning procedure - Ensure that all bags are clean before
using them for sampling. First, flush each bag three times with high-
purity nitrogen (N2; 99.998%). Then fill each bag with N2 and analyze the
bag contents at the highest sensitivity setting using the same analytical
technique that will be used for analyzing samples. Before constructing the
calibration curve, analyze one analytical system blank each day by taking
the gas chromatograph through its analytical program with no sample
injection. Analyze an analytical system blank again if carryover between
samples is indicated. Other, less stringent, methods of cleaning and
analysis may be used at the risk of overlooking important contaminants.
An acceptable level of contamination will be a response less than five
times the instrument detection limit or half of the level of concern,
whichever is less. Repeat the nitrogen flush as necessary until the
acceptable level has been reached. No bag shall be used until it has been
satisfactorily cleaned.
6.2 Sample bag storage and transport procedures - To ensure sampling
integrity, perform sample recovery in a manner that prevents contamination of the
bag sample. Protect the bag from sharp objects, direct sunlight and low ambient
temperatures (below 0°C) that could cause condensation of any of the analytes.
Store the bag samples in an area that has restricted access to prevent damage to
or tampering with the sample before analysis. Analyze the bag samples within 72
hours of sample collection unless it can be shown that significant (>20%) sample
degradation does not occur over a longer period of sample storage. Upon
completion of the testing and sample recovery, check all the data forms for
completeness and the sample bags for proper identification. Store the bags in
rigid, opaque containers during all sampling, storage and transport procedures.
Ship the bags using ground transportation. Follow all hazardous materials
shipping procedures.
6.3 Condensate storage and transport procedures - To ensure sampling
integrity, perform sample recovery in a manner that prevents contamination of the
condensate (Sec. 7.6.5). Store the condensate in 40-mL vials with no headspace.
Place the vials in ice or in a refrigerated container at 4°C (± 2°C) immediately
following recovery and during transport for analysis. In addition, store the
vials in an area that has restricted access to prevent damage to or tampering
0040 - 8 Revision 0
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with the sample before analysis. Upon completion of the testing and sample
recovery, check all the data forms for completeness and the condensate samples
for proper identification. Ship the condensate samples using ground
transportation. Follow all hazardous materials shipping procedures.
6.4 The time lapse between sampling and analysis shall not exceed 72 hours
unless it can be justified by specific sample matrix stability data that meets
the criteria of Sec. 1.5.3. Stability in a Tedlar® bag shall be demonstrated by
spiking analytes into inert gas in the laboratory and into stack gas in the
field. The spiking level must be at least at the level found in the samples of
the emissions matrix obtained during the pre-site survey. Compound recovery in
both laboratory and field studies must be > 80% after 72 hours for consideration
of applicability.
7.0 PROCEDURE
The overall sampling procedure involves a pretest survey of the source to
establish sampling parameters, a series of pretest checks of the sampling system
and the source conditions, and the actual sample collection. These steps are
described in Sees. 7.1 - 7.5. Following the actual sample collection step,
sampling data are recorded and a post-test leak check is performed (Sec. 7.6).
As noted in Sec. 1.0, this method does not include sample analysis procedures,
but general guidelines for sample analysis are given in Sec. 7.7. Sec. 7.8
provides an extensive set of calculations associated with the sample collection
and analysis procedures.
7.1 Pretest survey
7.1.1 Perform a pretest survey for each source to be tested. The
purpose of the survey is to obtain source information to select the
appropriate sampling and analysis parameters for that source. Potential
interferences may be detected and resolved during the survey. When
necessary information about the source cannot be obtained, collection and
analysis of actual source samples may be required.
7.1.2 The following information must be collected during a survey
before a test can be conducted. The information can be collected from
literature surveys and source personnel, but an actual on-site inspection
is recommended. A copy of the survey results must be forwarded to the
staff performing the sample analyses.
7.1.2.1 Determine whether the sampling site is in a
potentially explosive atmosphere. If the sample site is located in
an explosive atmosphere, use other, intrinsically safe test methods.
This method is never to be used in a potentially explosive atmosphere
(Sec. 1.2).
7.1.2.2 Measure and record the stack dimensions on a data
sheet similar to the data sheet shown in Figure 6. Select the
sampling site and the gaseous sampling points according to EPA Method
1 (Reference 9) or as specified by the regulatory personnel.
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7.1.2.3 Determine the stack pressure, temperature, and the
range of velocity pressures using EPA Method 2 (Reference 9).
7.1.2.4 Determine the stack gas moisture content
(Sec. 7.2.3) using EPA Approximation Method 4 (Reference 9) or its
alternatives. Perform the determination when process operations are
as they will be during final sampling. If the process uses and emits
ambient air, use a sling psychrometer to measure the moisture content
of the ambient air in the area of process air uptake.
7.1.2.5 Select a condensate collection system with a minimum
volume of 50 ml. Select a sampling rate and volume that will yield
a total condensate catch at or below 50 ml, to allow recovery of the
condensate into volatile organic analysis (VOA) vials with minimum
dead space.
7.1.2.6 In accordance with EPA Method 1, select a suitable
probe liner and probe length as determined by the temperature and
dimensions of the source. Determine the point within the stack that
represents an average flow and temperature of the stack. Mark the
probe at the determined distance to provide a reference point. For
sample collection, insert the probe into the duct to the predetermined
point to ensure proper probe placement and collection of a
representative sample.
7.1.2.7 Determine whether the source has a constant or
variable gas flow rate. The flow rate may be considered constant if
the variation over the sampling period is no more than 20%. If the
process is constant, use a constant sampling rate (Sec. 7.5.1). If
the process is not constant, use proportional sampling (Sec. 7.5.2).
7.1.2.8 Determine approximate levels of target compounds by
collecting a pretest bag sample for analysis. This information is
needed to establish parameters for the analytical system.
7.1.2.9 Check the sampling site to ensure that adequate
electrical service is available.
7.1.2.10 Follow all guidelines in the health and safety plan
for the test. Use appropriate safety equipment as required by
conditions at the sampling site (e.g., respirator, ear and eye
protection, and a safety belt).
7.2 Pretest procedures
7.2.1 Assemble the train according to the diagram in Figure 2.
Adjust the probe, filter, and valve heater controls to maintain a
temperature between 130°C and 140°C. Circulate cooling water from an ice
bath to the condenser until the temperature is stabilized at or below 20°C.
Allow the probe, filter, valve, and condenser temperatures to stabilize
before sampling. Mark the probe, pitot tube, and thermocouple assembly
with the proper sampling points as determined in accordance with EPA
Method 1. Before sampling, insert the pitot tube and thermocouple probe
into the stack, to allow the thermocouple readings to stabilize.
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7.2.2 Preliminary velocity and temperature traverse - While the
probe, filter, valve, and condenser temperatures are stabilizing, perform
a preliminary velocity/temperature traverse in accordance with EPA Methods
1 and 2. Record the velocity UP) and temperature (T,°C) at each point to
determine a point of average flow and velocity and measure the static
pressure at that point. Determine the average velocity head (APavg) and
range of fluctuation.
7.2.3 Determination of moisture content - Determine the moisture
content of the gas stream being sampled before (Sec. 7.1.2.4) or during
actual sampling. For combustion or water controlled processes (wet
electrostatic precipitators and scrubbers), obtain moisture content of the
flue gas during test conditions from plant personnel or by direct
measurement using EPA Method 4.
7.2.4 Criteria for selection of sample volume and flow rate - The
flow rate should fill the bag to at least 80% of its capacity during the
sampling period. The following criteria should be met:
7.2.4.1 Minimum stack sampling time for each run should be
1 hr. Data from less than 1 hr of sample collection would be an
invalid test run. Two hours of stack sampling time is recommended as
optimal.
7.2.4.2 The minimum sample volume shall be at least 15 L.
7.2.4.3 The minimum sample flow rate shall be 250 mL/min.
7.2.4.4 Typically, the average sampling flow rate is about
0.5 L/min, which will collect approximately 30 L of sample per hour.
7.2.4.5 Mass emission rate determination - Determine whether
the final result will be presented on a concentration or mass emission
basis before sampling. If results will be presented on a
concentration basis, only the concentrations of the target analytes
and the stack gas moisture content need to be measured. If the mass
emission rate of any compound is to be presented, the volumetric flow
rate of the stack gas must also be determined. The volumetric flow
rate may be determined by performing a temperature and velocity
traverse in accordance with EPA Methods 1 and 2, with actual sample
collection.
7.3 Leak check procedures
7.3.1 Bag evacuation and bag leak check procedure - Before sampling,
ensure that the Tedlar® bag is fully evacuated and leak free.
7.3.1.1 Assemble the sample train as illustrated in Figure
2 and described in Sec. 4.1.1, ensuring that all connections are
tight.
7.3.1.2 Turn the probe isolation valve to position 1 and
turn the bag isolation valve to position 1 (Figure 4).
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7.3.1.3 Disconnect the vacuum line from the bag container
(the quick connect has a valve to seal the line; Figure 2) and turn
on the pump in the control console (Figure 5).
7.3.1.4 Open the coarse adjustment valve and adjust the fine
adjustment valve on the control console (Figure 5) until the vacuum
gauge reads 5 in. Hg.
7.3.1.5 Turn the bag isolation valve to position 3
(Figure 4) and open the coarse valve completely to obtain maximum flow
rate.
7.3.1.6 Observe the dry gas meter and rotometer as the bag
is evacuated. The bag is completely evacuated when no flow is
indicated on the dry gas meter and the vacuum rises to 5 in. Hg
(Figure 5).
7.3.1.7 Allow the rotometer float ball to drop to zero.
Time and record the leak rate using one of the following procedures.
7.3.1.7.1 Timed leak rate (measured in liters per
minute) - Observe the leak rate indicated on the dry gas
meter and time for 1 min. The leak rate must be less than 4%
of the sample rate (e.g., 0.02 Lpm for a sample rate of 1
Lpm).
7.3.1.7.2 Timed pressure loss rate (measured in
inches Hg drop per minute) - Close both the coarse and fine
adjustment valves and turn off the pump. Observe the vacuum
gauge and time the pressure drop. The leak rate must be less
than or equal to 0.1 in. Hg/min.
7.3.1.8 If all connections are found to be leak tight and
the leak rate cannot meet the set criteria, discard the bag and test
another clean bag.
7.3.1.9 Turn the bag isolation valve to position 1 (Figure
4) to seal the evacuated bag.
7.3.1.10 Turn off the pump and turn the probe isolation valve
to position 3 (Figure 4) allowing the train to return to ambient
pressure.
7.3.1.11 Return the probe isolation valve to position 1, seal
the end of the probe and reconnect the vacuum line to the bag
container (Figures 2 and 4).
7.3.2 Pretest leak check
7.3.2.1 Before sampling and immediately after evacuating and
leak checking the bag, perform a pretest leak check of the sampling
train.
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7.3.2.2 Ensure that the bag isolation valve is in position
1 (Figure 4) and the end of the probe is sealed.
7.3.2.3 Turn the probe isolation valve to position 2 (Figure
4), turn the pump on, and open the coarse adjustment valve (Figure 5).
7.3.2.4 Allow the sample train to evacuate and adjust the
fine adjustment valve to increase the vacuum to 5 in. Hg (Figure 5).
7.3.2.5 When the rotometer drops to zero and the dry gas
meter slows to a stop, time and record the leak rate following the
procedures outlined in Sec. 7.3.1.7.
7.3.2.6 If the leak rate is greater than 0.1 in Hg/min or 4%
of the sampling rate, check all connections, valves, and the probe
seal for tightness. Any leak found must be corrected and the leak
check repeated before sampling collection begins.
7.3.2.7 After completing a satisfactory leak check, return
the sampling train to ambient pressure by turning the probe isolation
valve to position 3 (Figure 4) and turning off the pump (Figure 4).
7.3.2.8 When the vacuum gauge drops to zero, immediately
turn the probe isolation valve to position 1 (Figure 4).
7.3.3 Post-test leak check
7.3.3.1 A post-test leak check must be performed after each
bag sample is collected, before changing the bag and container for the
next sample.
7.3.3.2 Ensure that the bag and probe isolation valves are
in position 1 (Figure 4) and the pump is turned off when sample
collection is completed.
7.3.3.3 Remove the probe from the stack and seal the end of
the probe with a leak-tight seal. Check all connections and train
components for looseness or breakage. Do not tighten any connections.
Record any abnormal conditions.
7.3.3.4 Turn the probe isolation valve to position 2 (Figure
4) and disconnect the quick connectors on the bag isolation valve
return line from the tee on the vacuum line (Figure 2).
7.3.3.5 Turn on the pump and adjust the fine adjustment
valve until the train vacuum reaches at least 1 in. Hg above the
highest vacuum attained during sample collection. Time and record the
leak rate as previously outlined in Sec. 7.3.1.7.
7.3.3.6 If the leak rate is less than 4% of the sample rate
or 0.1 in. Hg/min., the sample is considered valid (Sees. 7.3.1.7.1
and 7.3.1.7.2).
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7.3.3.7 Return the sample train to ambient pressure
(Sees. 7.3.2.7 and 7.3.2.8) and disconnect the sample and vacuum lines
from the bag and container to prepare the train for the next sample.
7.3.3.8 If the post-test leak check proves invalid, discard
the invalid sample. Attach a new Tedlar® bag, evacuate and leak check
the bag, and repeat the sample collection.
7.4 Preparation for sample collection
7.4.1 Perform the pretest leak checks outlined in Sec. 7.3.
7.4.2 Remove the seal from the end of the probe and insert the probe
into the stack to the point of average velocity and temperature and
constant flow.
7.4.3 Purge the sampling train (probe, valve, and filter assembly
ONLY) using the following procedures.
7.4.3.1 Cap the inlet side of the charcoal purge trap
connected to the probe isolation valve tee using a 1/4 in. cap and
plug with Teflon® ferrules for an air-tight seal (Figure 2).
7.4.3.2 Disconnect the vacuum line quick connect from the
rigid bag container (the quick connect has a valve to seal the line).
7.4.3.3 Disconnect the return line connected to the bag
isolation valve from the quick connect at the vacuum line tee
(Figure 2).
7.4.3.4 Connect the purge line from the probe isolation
valve tee to the vacuum line tee using the quick connects (Figure 2).
7.4.3.5 Ensure that the bag isolation valve is in position
1 (Figure 4), turn on the pump, and turn the probe isolation valve to
position 2 (Figure 4).
7.4.3.6 Draw at least eight times the sample volume of flue
gas, or purge for at least 10 minutes, whichever is greater.
NOTE: A three-way valve may be used in place of the purge quick
connects at the vacuum line tee.
7.4.4 Adjust the sample flow rate to the desired setting and check
all temperature and flow readings during the purge to ensure proper
settings.
7.4.5 Purge the sampling train before and between the collection of
each sample during the test run.
7.4.6 Label each bag/container and VGA vial clearly, uniquely, and
consistently with its corresponding data form and run. Follow appropriate
traceability requirements as defined by the regulatory personnel.
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7.5 Sample Collection
Start sample collection after the pretest leak check (Sec. 7.3.2) and the
system purge (Sec. 7.4). Collect the sample using proportional rate sampling if
the pretest survey measurements (Sec. 7.1.2.7) show that the emission flow rate
varies by more than 20% over the sampling period. Otherwise, use constant rate
sampling. Prepare for sample collection for either method by turning the bag
isolation valve to position 2 (Figure 4) while the pump is still running from the
system purge.
If a viewing port has been incorporated in the bag container design,
visually inspect the Tedlar® bag frequently during the sampling run to ensure
that it is filling properly and that a sufficient sample volume is collected.
This frequent inspection will also help prevent overfilling and bursting the bag
during sampling.
7.5.1 Constant rate sampling
7,5.1.1 Place the end of the probe at a point within the
duct determined to have the average velocity and temperature and a
constant flow rate.
7.5.1.2 Record the start volume from the dry gas meter and
begin timing the sample period.
7.5.1.3 Take flue gas velocity and temperature readings
using either EPA Method 2A for smaller ducts (<24 inches) with a
remote pi tot tube and thermocouple or EPA Method 2 for larger ducts
(>24 inches). Utilizing a sample probe with pitot tubes and
thermocouples attached will generally ease sampling and will provide
a direct means to monitor flue gas velocity and temperature at the
sample probe inlet.
7.5.1.4 Record all required data upon starting, and at
intervals of no more than 5 minutes on the field sampling data form
(Figure 7).
7.5.1.5 Adjust the sample flow rate and sampling train
heating systems to the correct levels, after every velocity and
temperature reading. The tester must closely monitor the sample train
and control console to ensure that the sample flow rate does not vary
by more than 20% during any 5-minute period.
7.5.2 Proportional sampling
7.5.2.1 Position the probe in the center of the stack.
7.5.2.2 Record the start volume from the dry gas meter and
begin timing the sample period.
7.5.2.3 Monitor the velocity head during sampling as
described in Sec. 4.1.5 and maintain a constant proportion between the
sample flow rate and the flow rate in the duct. The flow rate to be
used during sampling (Sec. 7.2.2) is calculated using the proportional
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sample rate equation in Sec. 7.8.4. With this equation and the sample
rate assigned to the average flow rate, the rotometer setting can be
determined after each velocity reading and the sample rate set
accordingly.
7.5.2.4 Record all required data upon starting, and at
intervals of no more than 5 minutes on the field sampling data form
(Figure 7).
7.5.3 Single-point sampling
Collect samples from a single point within the duct as described in
Sees. 7.5.1.1 and 7.5.1.2, unless multipoint sampling has been determined
necessary (Sec. 7.5.1.4).
7.5.4 Multipoint sampling
Perform multipoint integrated sampling only in a case where there is
a possibility of effluent stratification. Stratification of gases is less
likely than of particulates. If however, multipoint sampling is required,
determine the necessary number of sample points in accordance with EPA
Methods 1 and 2.
7.6 Post-test procedures
7.6.1 Record the final volume from the dry gas meter at the end of
each sample collection period.
7.6.2 Perform a post-test leak check as described in Sec. 7.3.3.
7.6.3 Inspect the field sampling data form (Figure 7) and sample
identification labels for accuracy and completeness.
7.6.4 Replace the particulate filter after each sample.
7.6.5 Condensate Recovery - The condensate collected during sampling
must be recovered separately for each individual bag sample collected,
using the following procedures.
7.6.5.1 Carefully remove the condensate trap, the condenser
and the sample line (from the trap to the bag) from the sample train.
Pour the contents of the condensate trap into a clean measuring
cylinder.
7.6.5.2 Rinse the condenser, the condensate trap and the
sample three times with 10 ml of HPLC grade water and add the rinsings
to the measuring cylinder containing the condensate. Record the final
volume of the condensate and rinse mixture on the field sampling data
form (Figure 7). High moisture sources (such as those with wet
control devices) may require a 150-mL or 200-mL measuring cylinder
while low moisture sources (such as some rotary kilns and pyrolytic
incinerators) may require only a 100-mL size.
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7.6.5.3 Pour the contents of the measuring cylinder into a
20- or 40-mL amber glass VOA vial with a Teflon® septum screw cap.
Fill the vial until the liquid level rises above the top of the vial
and cap tightly. The vial should contain zero void volume (i.e., no
air bubbles). Discard any excess condensate into a separate container
for storage and transport for proper disposal.
7.6.5.4 Label each vial by using wrap around labels. Labels
can be preprinted or can be filled out on site.
7.7 Analytical Approach
The following description provides general guidelines to the analytical
approach rather than a comprehensive analytical protocol. The primary analytical
tool recommended for the measurement of volatile organic compounds in source
emissions is GC/MS using fused-silica capillary GC columns such as described in
Method 8260. Prescreening of the sample by gas chromatography with either flame
ionization (GC/FID) or, for electronegative compounds, electron capture detection
(GC/ECD) is recommended because it may not only be cost effective, but will also
yield information regarding the complexity and concentration of the sample. If
the smallest feasible injection loop saturates the analytical system, dilutions
of the sample can be made into Tedlar® bags using pure N2 (99.998%) as diluent.
Calculate the concentration of the volatile organic compounds in the gaseous
emissions by using the equations (14-18) in Sec. 7.8.10.
7.7.1 Analysis of gaseous components - Introduce the gases into the
gas chromatograph through the use of a sample loop. Use a cryogenic trap
if sample concentration before analysis if necessary.
For most purposes, electron ionization (El) mass spectra will be
collected because a majority of the volatile organic compounds give
characteristic El spectra. Also, El spectra are compatible with the NIST
Library of Mass Spectra and other mass spectral references, which aid in
the identification process for other components in the incinerator process
streams.
To clarify some identifications, chemical ionization (CI) spectra
using either positive ions or negative ions can be used to elucidate
molecular-weight information and simplify the fragmentation patterns of
some compounds. In no case, however, should CI spectra alone be used for
compound identification. For descriptions of GC conditions, MS conditions,
internal standard usage, and quantitative and quantitative identification,
refer to Method 8260,
7.7.2 Analysis of condensates - Refer to Method 5030 to analyze
condensate samples by using the purge and trap technique or by direct
aqueous injection. Use direct solvent injection if an organic phase is
present distinct from the aqueous phase. Use dilution as necessary to
prevent saturation of the analytical system.
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7.8 Calculations
7.8.1 Carry out all calculations for determining the concentrations
and emission rates of the target compounds. Round off figures to three
significant figures after final calculations.
7.8.2
A
AB
Ac
AVO
AT
Emission
**stock
D»
eff(std|
Nomenclature
Stack/source cross sectional area, m2 (ft2)
Amount of volatile organic compound in bag (ng)
Amount of volatile organic compound in condensate (ng)
Amount of volatile organic compound in analytical sample (ng)
Total amount of volatile organic compound (ng), AB + Ac
Water vapor in the gas stream, proportion by volume
(x!00=%H20)
Type S pitot tube coefficient (nominally 0.84 ± 0.02),
dimensionless.
Concentration of volatile organic compound in emissions
(M9/L)
Concentration of volatile organic compound per volume sampled
(M9/L)
Concentration of spiking standard in the Tedlar® bag
Concentration of spike standard in the stack/audit cylinder.
Volumetric flow rate of exhaust gas, L/min, ft3/ro-
Pitot tube constant,
34.97m/sec
11/2
gmole
) (mmHg)
(K) (mmH,0)
85.49 ft/sec
Ib
1bmole
HinHg)
(°R)(inH20)
1/2
0040 - 18
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La = Maximum acceptable leakage rate for a leak check, either
pretest or following a component change; equal to 0.00057 L
(0.02 ft3/min) or 4% of the average sampling rate, whichever
is less.
LDLVO, = Lower detectable amount of volatile organic compound in
entire sampling train.
L, = Individual leakage rate observed during the leak check
conducted before to the "ith" component change (i = 1, 2,
3...n) L/min.
Lp = Leakage rate observed during the post-test leak check, L/min.
Max Massvol = Maximum allowable mass flow rate (g/hr [lb/hr]) of volatile
organic compound emitted from the combustion source.
Max Concvol = Maximum anticipated concentration of the volatile organic
compound in the exhaust gas stream, g/m3 (Ib ft3).
Md = Stack-gas dry molecular weight, g/g-mole (Ib/lb-mole).
Mfd = Dry mole fraction of the flue gas.
M8 = Wet molecular weight of the flue gas.
Mw = Molecular weight of water, 18.0 g/g-mole (18.0 Ib/lb-mole).
Pbar = Barometric pressure at the sampling site, mm Hg (in. Hg).
Pg = Flue gas static pressure, mm H20 (in. H20).
Pk = Specific gravity of mercury (13.6)
P8 = Absolute stack gas pressure, mm Hg (in. Hg).
Pstd = Standard absolute pressure, 760 mm Hg (29.92 in. Hg).
Qm = Average sampling rate, L/min.
Qs = Calculated sampling rate, L/min.
Qsd = Volumetric air flow rate, (L/min, ft3/min).
R = Ideal gas constant, 0.06236 mm Hg-m3/K-g-mole (21.85 in. Hg-
ft3/cR-lb-mole).
Tm = Absolute average dry gas meter temperature, °K (°R).
T. = Absolute average stack gas temperature, °K (°R).
Tstd = Standard absolute temperature, 293°K (528°R).
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V;
V, cone
V
m(std)
U
* spike
VT
"w(std|
WF
Y
AH
AP
AP
p*
9
avg
Analytical sample volume (ml).
Bag volume (ml).
Concentration of volatile organic compound (wt %) introduced
into the combustion process.
Anticipated concentration of the volatile organic compound
in the exhaust gas stream, g/L (lb/ft3).
Total volume of liquid collected in the condensate knockout
trap.
Volume of gas sample as measured by dry gas meter, L.
Volume of gas sample measured by dry gas meter, corrected to
standard conditions, L.
Volume of gaseous or liquid spiking standard (ml)
Minimum dry standard volume to be collected at dry gas meter.
Train sample volume (ml)
Volume of water vapor in the gas sample, corrected to
standard conditions, L (ft3).
Stack gas velocity, calculated by Method 2, Equation 2-9,
using data obtained from Method 5, m/sec (ft/sec).
Mass flow rate of waste feed per hour, g/hr (Ib/hr).
Dry gas meter calibration factor, dimensionless.
Average pressure differential of orifice meter, inches H20.
Actual velocity pressure, mm (in.) H20.
Average velocity pressure, mm (in.) H20.
Density of water, 0.9982 g/mL (0.002201 Ib/mL).
Total sampling time, min.
Sampling time interval from the beginning of a run until the
first component change, min.
Sampling time interval between two successive component
changes, beginning with the interval between the first and
second changes, min.
0040 - 20
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0p = Sampling time interval from the final (nth) component change
until the end of the sampling run, min.
60 = Second/minute conversion.
100 = Percent conversion.
7.8.3 Conversion factors
To convert cubic feet (ft3) to liter (L), multiply by 0.02832.
7.8.4 Proportional sample rate calculation
The flow rate to be used during sampling when the velocity head varies
from the average is calculated using the following equation:
Q .,
M« Mm
7.8.5 Dry gas volume - Correct the sample measured by the dry gas
meter to standard conditions (20"C, 760 mm Hg) by using the following
equation:
T.. Pk + AH/13.6 PK + AH/13.6
V = V / — bar = K V v b"
m(std) m' j p *1 m' -r
1 m std m
where:
K, = 0.3858 K/mm Hg for metric units, or
K, = 17.64°R/in. Hg for English units.
Equation 2 can be used as written, unless the leakage rate observed
during any of the mandatory leak checks (i.e., the post-test leak check or
leak checks conducted before component changes) exceeds La. If Lp or L|
exceeds La, Equation 2 must be modified as follows (with the approval of
the appropriate regulatory personnel):
7.8.5.1 Case I (no component change during sampling run)
Replace Vm in Equation 2 with the expression:
0040 - 21 Revision 0
January 1995
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7.8.5.2 Case II (one or more component changes during the
sampling run)
Replace Vm in Equation 2 with the expression:
V -
m
1=1
and substitute only for those leakage rates (Lj or Lp) that exceed La
7.8.6 Volume of water vapor
V = V —
w(std) "lc u
K2V1c
w std
where:
K2 = 0.001333 m3/mi for metric units, or
K2 = 0.04707 ft3M for English units.
7.8.7 Moisture content
V
B
w (std)
WS V + V
m (std) w (std)
i
7.8.8
Volumetric flow rate equations
7.8.8.1 Static pressure
P
P = Pn +
s Bar
7.8.8.2 Dry molecular weight
Md = (% C02 x 0.44) + (% 02 x 0.32) + [(% CO + % N2) x 0.28]
7.8.8.3 Dry mole fraction
0040 - 22
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7.8.8.4 Wet molecular weight
M = (M. x M,.) + (18 x B )
8 x d fd' v ws'
7.8.8.5 Flue gas velocity
v = k c
avg
M P
s s
7.8.8.6 Volumetric flow rate
DV.ff(std, - 60 Vs
fd
std
8 (avg)
std
7.8.9 Concentration of a volatile organic compound in the gaseous
emissions of a combustion process
7.8.9.1 Divide the amount of volatile organic compound
determined through analysis by the volume of sample introduced into
the analytical system to obtain concentration of the volatile organic
compound in the bag or the condensate.
vol
vol
7.8.9.2 Multiply the concentration of the volatile organic
compound (ng/l) by the sample volume (bag or condensate) to determine
the amount of the volatile organic compound in the bag or condensate.
or
= C , x V0
vol B
A = C , x V,
C vol Ic
7.8.9.3 Sum the amount of volatile organic compound found in
all samples associated with a single train.
AT = AB
7.8.9.4 Divide the total amount found by the volume of stack
gas sampled to determine the concentration of the volatile organic
compound in the gaseous emissions.
0040 - 23
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T
-I = Cc . .
if Emission
VT
7.8.10 Concentration of the spiking standard in the Tedlar® bag
V X C
r- _ spike stock
spike y
VB
7.8.11 Recovery of the spiking standard from the Tedlar® bag sample
% Recovery = _J£L X 100
spike
8.0 QUALITY CONTROL
8.1 Quality assurance/quality control requirements before sampling
8.1.1 Pitot tube probe - Before sampling, assemble and calibrate the
pitot tube probe (described in Sec. 4.2.11) in accordance with EPA
Method 2. Leak check to ± 10 in. H20. The pitot tube assembly must be
leak free (0.00 in. H20 in 1 minute).
8.1.2 Pressure gauge (manometer) - Calibrate the pressure gauge
(described in Sec. 4.2.12) in accordance with EPA Method 2. Leak check the
pitot tubes, pressure gauge, and pitot tube lines simultaneously, as a
unit, before the velocity traverse.
8.1.3 Thermocouple and temperature read-out device - Calibrate these
devices (described in Sec. 4.2.10.6) within 30 days of sampling and in
accordance with EPA Method 2. The thermocouple and temperature read out
must be accurate to ± 1"C.
8.1.4 Metering system - Calibrate the dry gas meter contained in the
control console in accordance with the procedures outlined in Sec. 5.3 of
EPA Method 5. Calibrate the meter at a flow rate appropriate for the
sampling rate used during the test.
8.1.5 Probe heater - Calibrate the probe heater before sampling
collection following procedures outlined in Sec. 5.5 of EPA Method 5.
8.1.6 Barometer - Adjust the barometer daily and before each test
series to ± 0.1 in. (25 mm) Hg of the corrected barometric pressure
reported by a National Weather Service Station located nearby and at the
same altitude above sea level.
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8.2 Number of sampling runs
The number of sampling runs to be performed shall be determined by the
appropriate regulatory personnel. At least two runs (two hours of stack sampling
time) are recommended for each test series to provide minimal statistical data.
Ensure that all compounds on the analyte list have been validated for this
method prior to sampling. Perform validation as required in accordance with EPA
Method 301 (Reference 9).
8.3 Blanks and field spikes
Field, trip and laboratory blanks, contamination checks and field spiked
samples are required to monitor the performance of the sampling method and to
provide the required information to take corrective action if problems are
observed in the laboratory operations or in field sampling activities.
8.3.1 Field blanks - Take at least one field blank sample daily and
per source. Collect high purity air or N2 (99.998%) from a compressed gas
cylinder in the same manner as source emissions. Draw the air or nitrogen
gas through the sampling system and into the bag. Field blank samples
shall consist of the condensate and a bag sample. Transport and analyze
this blank sample along with the stack gas samples. When the field blank
values are greater than 20% of the stack values, flag the data. Report the
field blank values with the stack gas results.
8.3.2 Trip blanks - Take at least two Tedlar® bags labeled "trip
blanks" and filled with an inert gas to the sampling site. These bags will
be treated like any other samples except that they will not be opened
during storage at the site. These bags will be subsequently analyzed to
monitor potential contamination which may occur during storage and
shipment.
8.3.3 Laboratory blanks - Leave two Tedlar® bags labeled "laboratory
blanks" in the laboratory using the method of storage that is used for
field samples. If the field and trip blanks contain high concentrations
of contaminants (i.e., greater than five times the detection limit of
particular analyte), the laboratory blank shall be analyzed to identify the
source of contamination.
8.3.4 Tedlar® bag contamination checks - The use of new bags for
each test series is recommended. All bags must be cleaned and checked for
contamination before being used for sampling (Sec. 6.1.3).
8.3.5 Field spike samples - Take at least one field spike sample per
10 field samples, or a minimum number of one field spike per test. Spike
the chosen bag sample with a known mixture (gaseous or liquid) of all the
target pollutants using either gaseous or liquid injection into the bag.
Transport and analyze the spiked sample with the stack gas samples. Report
the spike sample recoveries with the source test results. The compound
recoveries in the spiked sample must be 80 - 120%. Use Equation 17 in Sec.
7.8.11 to calculate spiking compound recovery.
0040 - 25 Revision 0
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The spiking concentration should be at least the concentration
anticipated in the emissions matrix. Use Equation 16 in Sec. 7.8.10 to
calculate the spiking concentration. The syringe volume for the gaseous
injection should not exceed 200 ml to minimize leakage through the septum
after injection. For liquid injections, the volume injected must not
exceed 1 ml to ensure complete volatilization. The final volume of the
spiked gas must not exceed 1% of the total sample volume. Use the ideal
gas equation to calculate the volume of gas generated by a liquid injection
into the bag.
8.3.5.1 Obtain spiking stock that is sufficiently
concentrated to spike a Tedlar® sample without exceeding 1% volume
limit. Select appropriate analyzes, analyte homologs, or isotopically
labeled analogs in cylinders or SUMMA® canisters for gaseous
injections or neat liquids or methanol solutions for liquid
injections.
8.3.5.2 Install an injection port that consists of a
Swagelok® tee fitting with a septum, in the sample line just before
the 1/4-in. quick connector on the Tedlar® bag (Figure 2). Locate
this port as close to the bag as possible to minimize wall effects.
Use a new septum for each sampling run that involves spiking.
8.3.5.3 Perform a leak test as described in Sec. 7.3 with
the injection port in line.
8.3.5.4 Start sampling the stack as described in Sees. 7.4
and 7.5.
8.3.5.5 In preparation for injection, clean the syringe by
flushing three times with an inert gas (high purity N2, 99.998%) for
gaseous injections, or with methanol for liquid injections. Then
flush the syringe three times with the gaseous or liquid spiking
standard.
8.3.5.6 After half an hour of sample collection, take up the
desired volume of the spiking standard into the syringe (for gases,
allow the standard to equilibrate to atmospheric pressure) and inject
it through the septum into the bag without interrupting the sampling
procedure. All apparatus upstream of the bag should be under slight
negative pressure.
8.4 Performance audits should be conducted to evaluate quantitatively the
quality of data produced by the total measurement system (sample collection,
sample analysis, and data processing). Accuracy (% recovery) must be 50 - 150%.
Precision (% relative standard deviation) must be less than or equal to
50 percent.
8.5 Evaluation of analytical procedures for a selected series of compounds
shall include the sample preparation procedures and each associated analytical
determination. Challenge the analytical procedures by spiking the test compounds
at appropriate levels carried through the procedures.
0040 - 26 Revision 0
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8.6 Determine the overall method detection limits (lower and upper) on a
compound-by-compound basis according to the 40 CFR Part 136b for the
determination of the detection limit. Different compounds may exhibit different
collection efficiencies as well as instrumental minimum detection limit.
8.7 Determine the overall method precision and bias (in accordance with
the EPA Method 301) on a compound-by-compound basis at a given concentration
level. Include in the method precision value a combined variability due to
sampling and instrumental analysis. The method bias is dependent upon the
collection efficiency of the train components.
9.0 METHOD PERFORMANCE
No evaluation or validation data are currently available for this method.
10.0 REFERENCES
1. Howe, G.B., Pate, B.A., and Jayanty, R.K.M., "Stability of Volatile
Principal Organic Hazardous Constituents (POHCs) in Tedlar® Bags", Research
Triangle Institute Report to the EPA, Contract No. 68-02-4550, 1991.
2. Andino, J.M., and Butler, J.W., "A Study of the Stability of Methanol-
Fueled Vehicle Emissions in Tedlar® Bags", Environ. Sci. Technol. 1991,
25(9), 1644-1646.
3. Posner, J.C., and Woodfin, W.J., "Sampling with Gas Bags I: Loses of
Analyte with Time", Appendix L Industrial Hygiene, 1986, (4), 163-168.
4. Seila, R.L., Lonneman, W.A., and Meeks, S.A., "Evaluation of Polyvinyl
Fluoride as a Container Material for Air Pollution Samples", J. Environ.
Sci. Health., 1976, 2, 121-130.
5. U.S. Environmental Protection Agency, Hazardous Waste Incineration
Measurement Guidance Manual, Volume III of the Hazardous Waste Incineration
Guidance Series, EPA/625/6-89/021, p. 5.
6. U.S. Environmental Protection Agency, Method 301, "Protocol for the Field
Validation of Emission Concentrations from Stationary Sources", EPA 450/4-
90-015, February 1991.
7. U.S. Environmental Protection Agency, 40 CFR Part 136, Appendix B,
"Definition and Procedure for the Determination of the Method Detection
Limit".
8. Kanniganti, R., Moreno, R.L., and Bursey, J.T., Radian Corporation,
Research Triangle Park, North Carolina, "Method 0040: Sampling of Principal
Organic Hazardous Constituents from Combustion Sources Using Tedlar® Bags",
EPA Contract No. 68-D1-0010.
9. U.S. Environmental Protection Agency, 40 CFR Part 60, Appendix A,
Methods 1, 2, 3, 4, 5, 18 and 25.
0040 - 27 Revision 0
January 1995
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TABLE 1
PROBLEMS THAT CAN INVALIDATE TEDLAR® BAG SAMPLING DATA AND SUGGESTED REMEDIES
Problem
Remedy
1. Condensation of the gases or
water vapor in the bag
2. Leaks developing in the bag
during testing, transport,
and/or analysis
3. Hydrocarbon contamination
Sample below the condensation point
of the analytes; lower the
temperature in the condensate trap.
Use double sealed bags; perform
additional sampling runs; protect the
bags from sharp objects by sampling
and shipping in rigid, opaque
containers; ship the bags in the same
containers used during sampling.
Minimize exposure of the bag to heat
and direct light, by sampling and
shipping in rigid, opaque containers;
purge the bags with ultrapure N2 in
the laboratory and establish through
analysis that the hydrocarbon levels
are acceptable; use the bags only
once.
0040 - 28
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January 1995
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<
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00
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w
c o>
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FIGURE 2
ISOLATION VALVE DESIGN
3-Way Glass Valve
With Teflon Stopcock
r
In
Valve Stem Reduced
From 5/16" to 1/4"
Out
*
1"-
(All 3 Stems Of Equal
Length, Size, & Shape)
Heat Wrap Valve
And Heat To 13(3-140° C
Out
0040 - 30
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January 1995
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FIGURE 3
VALVE OPERATION
Isolation Position
(Post Test)
Probe Isolation Valve Positions
@ Sample Position Q) Vacuum Purge Position
System Purge Position (Pre Test) (Release System Pressure
After Leak Checks)
u) Isolation Position
System Purge Position (Pre Test)
Leak Check Position (Post Test)
Bag Isolation Valve Positions
@ Sample Position
) Bag Evacuation Position
Bag Leak Check Position
(Pre Test)
0040 - 31
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January 1995
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FIGURE 4
DIAGRAM OF A CONTROL CONSOLE
Front View
(D
1234
LPM
Schematic Diagram
LOUT
0040 - 32
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January 1995
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FIGURE 4 (Continued)
CONTROL CONSOLE COMPONENTS
1. 1/4 in. S.S. Quick Connect - Vacuum line inlet from sample train (to bag
container).
2. Amphenol Receptacle - provides power through umbilical to probe heat &
water pump.
3. Thermocouple Receptacles - 4 thermocouple inlets for:
a. Stack Temperature
b. Probe Temperature
c. Condenser Temperature
d. Ambient Temperature
4. 110 VAC Receptacle - auxiliary power for isolation valve heat.
5. Vacuum Gauge - 0-30 in. Hg.
6. Heat Controller
7. Digital Thermocouple Read Out - 10 channel (displays temperature
readings during sampling)
(1-4 remote as listed above)
(5 dry gas meter temperature)
(6-10 spares)
8. Timer (optional)
9. Power Switches - control (on/off)
a. Main power - with separate switches for each.
b. Sample pump
c. Water pump
d. Timer
10. Meter pressure Gauge - (inches water column)
11. Fine Adjustment (Bypass) Valve
12. Coarse Adjustment (on/off) Valve
13. Dry Gas Meter
14. Rotometer (Flow Meter)
15. Charcoal Trap (Optional)
0040 - 33 Revision 0
January 1995
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FIGURE 5
PRETEST SURVEY DATA FORM
I. Name of Company
Address
Contacts
Phone Numbers
Date
Process to be sampled
Duct or vent to be sampled
II. Process description
Raw material
Products
Operating cycle
Check: Batch
Timing of batch or cycle
Best time to test
Continuous
Cyclic
III. Sampling site
Description
Site description _
Duct shape and size
Materials
Wall thickness
Upstream distance
Downstream distance
Size of port
inches
inches
inches
diameter
diameter
0040 - 34
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January 1995
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FIGURE 5 (Continued)
Temperature
Velocity _
Static pressure
Moisture content
Particulate content
Gaseous components
N,
0
2
CO
C02
so,
°c
01
h
01
/o
Hydrocarbon components
inches H20
Data Source
Data Source
Data Source
Data Source
Data Source
Hydrocarbons
ppm
ppm
ppm
ppm
ppm
ppm
ppm
ppm
ppm
ppm
ppm
B. Sampling considerations
Location to set up GC
Power available at duct
Plant entry requirements
Security agreements
Potential problems
Site diagrams (Attach additional sheets if required)
0040 - 35
Revision 0
January 1995
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METHOD 0040
SAMPLING OF PRINCIPAL ORGANIC HAZARDOUS CONSTITUENTS
FROM COMBUSTION SOURCES USING TEDLAR® BAGS
Applicable Source
and Analytes
Explosion Hazard Area
Compounds with Boiling
Points > 121 C
Compounds Unstable in Bag
J Use Other
*V Sampling Methods
Presurvey
Measurements
Select
Condenser Trap Size
Analytical Detector
Flow Rate (Proportional vs Constant)
Preparation
and Set-up
Assembly of the Sample Tram
Preliminary Velocity and Temperature Traverse
Determination ot Moisture Content
Selection of Sample Volume and Flow Rate
Bag
Leak Check
Leak Rate > 0 1 in Hg in 1 mm
or > 4% of the Sample Rate
Discard Bag
and Use Another
Acceptable Leak Rate
Pre-test Train
Leak Check
Leak Rate > 0 1 in. Hg in 1 mm.
or > 4% of the Sample Rate
Acceptable Leak Rate
Check All Connections
Repeat Leak Check
Insert Probe
into Gas Stream
0040 - 38
Revision 0
January 1995
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METHOD 0040 (Continued)
SAMPLING OF PRINCIPAL ORGANIC HAZARDOUS CONSTITUENTS
FROM COMBUSTION SOURCES USING TEDLAR® BAGS
Insert Probe
into Gas Stream
I
Purge
Probe Assembly
Collect Sample
Using Proportional
Sampling Rate
Steady
Flow Rate
Source
Collect Sample
Using Constant
Sampling Rate
Post-test Tram
Leak Check
Leak Rate > 0 1 in Hg in 1 mm.
or > 4% of the Sample Rate
Rag Invalid Test Data
Discard Invalid Samples
Repeat Test Run
Acceptable Leak Rate
Sample Recovery
and Transport
I
Post-transport
Leak Check on Bags
In-leakage > 20V.
Out-leakage > 20%
Flag Invalid Test Data >.
Discard Invalid Samples )
Acceptable Leakage
Analysis
(within 72 Hours of
Sample Collection)
0040 - 39
Revision 0
January 1995
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METHOD 0050
ISOKINETIC HC1/C1, EMISSION SAMPLING TRAIN
1.0 SCOPE AND APPLICATION
1.1 This method describes the collection of hydrogen chloride (HC1, CAS
Registry Number 7647-01-0) and chlorine (C12, CAS Registry Number 7782-50-5) in
stack gas emission samples from hazardous waste incinerators and municipal waste
combustors. The collected samples are analyzed using Method 9057. This method
collects the emission sample isokinetically and is therefore particularly suited
for sampling at sources, such as those controlled by wet scrubbers, emitting acid
particulate matter (e.g., HC1 dissolved in water droplets). A midget impinger
train sampling method designed for sampling sources of HC1/C12 emissions not in
particulate form is presented in Method 0051.
1.2 This method is not acceptable for demonstrating compliance with HC1
emission standards less than 20 ppm.
1.3 This method may also be used to collect samples for subsequent
determination of particulate emissions (SW-846 Method 0010) following the
additional sampling procedures described.
2.0 SUMMARY OF METHOD
2.1 Gaseous and particulate pollutants are withdrawn from an emission
source and are collected in an optional cyclone, on a filter, and in absorbing
solutions. The cyclone collects any liquid droplets and is not necessary if the
source emissions do not contain liquid droplets. The Teflon mat or quartz-fiber
filter collects other particulate matter including chloride salts. Acidic and
alkaline absorbing solutions collect gaseous HC1 and C12, respectively.
Following sampling of emissions containing liquid droplets, any HC1/C12 dissolved
in the liquid in the cyclone and/or on the filter is vaporized to gas and
ultimately collected in the impingers by pulling Ascarite IIR conditioned ambient
air through the sampling train. In the acidified water absorbing solution, the
HC1 gas is solubilized and forms chloride (CV) ions. The C12 gas present in the
emissions has a very low solubility in acidified water and passes through to the
alkaline absorbing solution where it undergoes hydrolysis to form a proton (H+),
CV' and hypochlorous acid (HC10). The (CV) ions in the separate solutions are
measured by ion chromatography (Method 9057). If desired, the particulate matter
recovered from the filter and the probe is analyzed following the procedures in
SW-846 Method 0010.
2.2 The stoichiometry of HC1 and C12 collection in the sampling train is
as follows: In the acidified water absorbing solution, the HC1 gas is
solubilized and forms chloride ions (CT) according to the following formula:
HC1 + H20 = H30+ + Cl"
0050 - 1 Revision 0
January 1995
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The C12 gas present in the emissions has a very low solubility in acidified water
and passes through to the alkaline absorbing solution where it undergoes
hydrolysis to form a proton (H+), Cl", and hypochlorous acid (HC10) as shown:
H20 + C12 = H+ + Cl" + HC10
3.0 INTERFERENCES
3.1 Volatile materials which produce chloride ions upon dissolution during
sampling are obvious interferences in the measurement of HC1. One interferant
for HC1 is diatomic chlorine (C12) gas which disproportionates to HC1 and
hypochlorous acid (HC10) upon dissolution in water. C12 gas exhibits a low
solubility in water, however, and the use of acidic rather than neutral or basic
solutions for collection of hydrogen chloride gas greatly reduces the dissolution
of any chlorine present.
4.0 APPARATUS AND MATERIALS
4.1 Sampling Train.
4.1.1 A schematic of the sampling train used in this method is shown
in Figure 1. This sampling train configuration is adapted from EPA Method
5 and SW-846 Method 0010 procedures, and, as such, the majority of the
required equipment is identical to that used in Method 0010
determinations. The new components required are a glass nozzle and probe,
a Teflon union, a quartz-fiber or Teflon mat filter (see Section 5.5), a
Teflon frit, and acidic and alkaline absorbing solutions.
4.1.2 Construction details for the basic train components are
provided in Section 3.4 of EPA's Quality Assurance Handbook, Volume III
(Reference 2); commercial models of this equipment are also available.
Additionally, the following subsections identify allowable train
configuration modifications.
4.1.3 Basic operating and maintenance procedures for the sampling
train are also described in Reference 2. As correct usage is important in
obtaining valid results, all users should refer to Reference 2 and adopt
the operating and maintenance procedures outlined therein unless otherwise
specified. The sampling train consists of the components detailed below.
4.1.3.1 Probe nozzle. Glass with sharp, tapered (30° angle)
leading edge. The taper shall be on the outside to preserve a
constant I.D. The nozzle shall be buttonhook or elbow design. The
nozzle should be coupled to the probe liner using a Teflon union.
It is recommended that a stainless steel nut be used on this union.
In cases where the stack temperature exceeds 210°C (410°F), a one-
piece glass nozzle/liner assembly must be used. A range of nozzle
sizes suitable for isokinetic sampling should be available. Each
nozzle shall be calibrated according to the procedures outlined in
Method 0010 sec. 9.1.
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4.1.3.2 Probe liner. Borosilicate or quartz-glass tubing with
a heated system capable of maintaining a gas temperature of 120 ±
14°C (248 ± 25°F) at the exit end during sampling. Because the
actual temperature at the outlet of the probe is not usually
monitored during sampling, probes constructed and calibrated
according to the procedure in Reference 2 are considered acceptable.
Either borosilicate or quartz-glass probe liners may be used for
stack temperatures up to about 480°C (900°F). Quartz liners shall
be used for temperatures between 480 and 900°C (900 and 1650°F).
(The softening temperature for borosilicate is 820°C (1508°F), and
for quartz 1500°C (2732°F).) Water-cooling of the stainless steel
sheath will be necessary at temperatures approaching and exceeding
500°C.
4.1.3.3 Pitot tube. Type S, as described in Section 2.1 of
EPA Method 2 (Reference 1). The pitot tube shall be attached to the
probe to allow constant monitoring of the stack-gas velocity. The
impact (high-pressure) opening plane of the pitot tube shall be even
with or above the nozzle entry plane (see Section 3.1.1 of Reference
2) during sampling. The Type S pitot tube assembly shall have a
known coefficient, determined as outlined in Section 3.1.1 of
Reference 2.
4.1.3.4 Differential pressure gauge. Inclined manometer or
equivalent device as described in Section 2.2 of EPA Method 2
(Reference 1). One manometer shall be used for velocity-head (delta
P) readings and the other for orifice differential pressure (delta
H) readings.
4.1.3.5 Cyclone (optional), glass.
4.1.3.6 Filter holder. Borosilicate glass, with a Teflon frit
filter support and a sealing gasket. The sealing gasket shall be
constructed of Teflon or equivalent materials. The holder design
shall provide a positive seal against leakage at any point along the
filter circumference. The holder shall be attached immediately to
the outlet of the cyclone.
4.1.3.7 Filter heating system. Any heating system capable of
maintaining a temperature of 120 ± 14°C (248 ± 25"F) around the
filter holder and cyclone during sampling. A temperature gauge
capable of measuring temperature to within 3°C (5.4°F) shall be
installed so that the temperature around the filter holder can be
regulated and monitored during sampling.
4.1.3.8 Impinger train. The following system shall be used
to determine the stack gas moisture content and to collect HC1 and
C12: five or six impingers connected in series with leak-free ground
glass fittings or any similar leak-free non-contaminating fittings.
The first impinger shown in Figure 1 (knockout or condensate
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impinger) is optional and is recommended as a water knockout trap
for use under test conditions which require such a trap. If used,
this impinger should be constructed as described below for the
alkaline impingers, but with a shortened stem, and should contain 50
ml of 0.05 M H2S04. The following two impingers (acid impingers
which each contain 100 ml of 0.05 M H2S04) shall be of the Greenburg-
Smith design with the standard tip (see Method 0010, Section
4.1.3.8). The next two impingers (alkaline impingers which each
contain 100 mL of 0.1 M NaOH) and the last impinger (containing
silica gel) shall be of the Greenburg-Smith design modified by
replacing the tip with a 1.3-cm (1/2-in.) I.D. glass tube extending
about 1.3 cm (1/2 in.) from the bottom of the impinger (see Method
5, Section 4.1.3.8).
The condensate, acid, and alkaline impingers shall contain known
quantities of the appropriate absorbing reagents. The last impinger
shall contain a known weight of silica gel or equivalent desiccant.
4.1.3.9 Metering system. The necessary components are a
vacuum gauge, leak-free pump, thermometers capable of measuring
temperature to within 3°C (5.4°F), dry-gas meter capable of
measuring volume to within 1%, an orifice meter (rate meter), and
related equipment, as shown in Figure 1. At a minimum, the pump
should be capable of 4 cfm free flow, and the dry-gas meter should
have a recording capacity of 0-999.9 cu ft with a resolution of
0.005 cu ft. Other metering systems capable of maintaining sampling
rates within 10% of isokineticity and of determining sample volumes
to within 2% may be used. The metering system must be used in
conjunction with a pitot tube to enable checks of isokinetic
sampling rates.
4.1.3.10 Barometer. Mercury, aneroid, or other barometer
capable of measuring atmospheric pressure to within 2.5 mm Hg (0.1
in. Hg). In many cases, the barometric reading may be obtained from
a nearby National Weather Service station, in which case the station
value (which is the absolute barometric pressure) is requested and
an adjustment for elevation differences between the weather station
and sampling point is applied at a rate of minus 2.5 mm Hg (0.1 in.
Hg) per 30-m (100 ft) elevation increase (vice versa for elevation
decrease).
4.1.3.11 Gas density determination equipment. Temperature
sensor and pressure gauge (as described in Sections 2.3 and 2.4 of
EPA Method 2), and gas analyzer, if necessary (as described in EPA
Method 3, Reference 1). The temperature sensor ideally should be
permanently attached to the pitot tube or sampling probe in a fixed
configuration such that the tip of the sensor extends beyond the
leading edge of the probe sheath and does not touch any metal.
Alternatively, the sensor may be attached just prior to use in the
field. Note, however, that if the temperature sensor is attached in
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the field, the sensor must be placed in an interference-free
arrangement with respect to the Type S pitot tube openings (see EPA
Method 2, Figure 2-7). As a second alternative, if the stack gas is
saturated, the stack temperature may be measured at a single point
near the center of the stack.
4.1.3.12 Ascarite tube for conditioning ambient air. Tube
tightly packed with approximately 150 g of fresh 8 to 20 mesh
Ascarite IIR sodium hydroxide coated silica, or equivalent, to dry
and remove acid gases from the ambient air used to remove moisture
from the filter and optional cyclone. The inlet and outlet ends of
the tube should be packed with at least 1 cm thickness of glass wool
or filter material suitable to prevent escape of Ascarite II fines.
Fit one end with flexible tubing, etc. to allow connection to probe
nozzle.
4.2 Sample Recovery.
4.2.1 Probe liner. Probe and nozzle brushes; nylon bristle brushes
with stainless steel wire handles are required. The probe brush shall
have extensions of stainless steel, Teflon, or inert material at least as
long as the probe. The brushes shall be properly sized and shaped to brush
out the probe liner and the probe nozzle.
4.2.2 Wash bottles. Two. Polyethylene or glass, 500 ml or larger.
4.2.3 Glass sample storage containers. Glass, 500- or 1,000-mL.
Screw-cap liners shall be Teflon and constructed so as to be leak-free.
Narrow-mouth glass bottles have been found to exhibit less tendency toward
leakage.
4.2.4 Petri dishes. Glass or plastic sealed around the circumfer-
ence with Teflon tape, for storage and transport of filter samples.
4.2.5 Graduated cylinder and/or balances. To measure condensed
water to the nearest 1 ml or 1 g. Graduated cylinders shall have
subdivisions not >2 ml. Laboratory triple-beam balances capable of
weighing to ± 0.5 g or better are required.
4.2.6 Plastic storage containers. Screw-cap polypropylene or
polyethylene containers to store silica gel.
4.2.7 Funnel and rubber policeman. To aid in transfer of silica gel
to container (not necessary if silica gel is weighed in field).
4.2.8 Funnels. Glass, to aid in sample recovery.
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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 reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Reagent water. All references to water in the method refer to reagent
grade water as defined in Chapter One unless otherwise specified. It is
advisable to analyze a blank sample of this water prior to sampling, since the
reagent blank values obtained during the field sample analysis must be less than
10 percent of the sample values.
5.3 Sulfuric acid (0.05 M), H2S04. Used as the HC1 absorbing reagent in
the impinger train. To prepare 1 L, slowly add 2.80 ml of concentrated H2S04 to
about 900 ml of water while stirring, and adjust the final volume to 1-L using
additional water. Shake well to mix the solution. It is advisable to analyze
a blank sample of this reagent prior to sampling, since the reagent blank values
obtained during the field sample analysis must be less than 10 percent of the
sample values.
5.4 Sodium hydroxide (0.1 M), NaOH. Used as the C12 absorbing reagent in
the impinger train. To prepare 1 L, dissolve 4.00 g of solid NaOH in about 900
mL of water and adjust the final volume to 1-L using additional water. Shake
well to mix the solution. It is advisable to analyze a blank sample of this
reagent prior to sampling, since the reagent blank values obtained during the
field sample analysis must be less than 10 percent of the sample values.
5.5 Filter. Quartz-fiber or Teflon mat (e.g., PallflexR TX40HI45) filter,
or equivalent.
5.6 Silica gel. Indicating type, 6-16 mesh. If previously used, dry at
175°C (350°F) for 2 hours before using. New silica gel may be used as received.
Alternatively, other types of desiccants may be used if equivalence can be
demonstrated.
5.7 Acetone. When using this train for determination of particulate
emissions, reagent grade acetone, < 0.001 percent residue, in glass bottles is
required. Acetone from metal containers generally has a high residue blank and
should not be used. Sometimes suppliers transfer acetone to glass bottles from
metal containers; thus, acetone blanks shall be run prior to field use and only
acetone with low blank values (< 0.001 percent) shall be used. In no case shall
a blank value greater than 0.001 percent of the weight of acetone used be
subtracted from the sample weight.
5.8 Crushed ice. Quantities ranging from 10-50 Ib may be necessary during
a sampling run, depending on ambient air temperature.
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5.9 Stopcock grease. Acetone-insoluble, heat-stable silicone grease may
be used, if needed. Silicone grease usage is not necessary if screw-on
connectors or Teflon sleeves on ground-glass joints are used.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Sample collection is described in this method. The analytical
procedures for HC1 and C12 are described in Method 9057 and for particulate
matter in Method 0100.
6.2 Samples should be stored in clearly labeled, tightly sealed containers
between sample recovery and analysis. They may be analyzed up to four weeks
after collection.
7.0 PROCEDURE
7.1 Preparation for Field Test.
7.1.1 All sampling equipment shall be maintained and calibrated
according to the procedures described in Section 3.4.2 of EPA's Quality
Assurance Handbook, Volume III (Reference 2).
7.1.2 Weigh several 200-300-g portions of silica gel in airtight
containers to the nearest 0.5-g. Record on each container the total
weight of the silica gel plus containers. As an alternative to
preweighing the silica gel, it may instead be weighed directly in the
impinger just prior to train assembly.
7.1.3 Check filters visually against light for irregularities and
flaws or pinhole leaks. Label the shipping containers (glass or plastic
Petri dishes) and keep the filters in these containers at all times except
during sampling (and weighing for particulate analysis).
7.1.4 If a particulate determination will be conducted, desiccate
the filters at 20 ± 5.6°C (68 ± 10'F) and ambient pressure for at least 24
h, and weigh at intervals of at least 6 h to a constant weight (i.e.,
<0.5-mg change from previous weighing), recording results to the nearest
0.1 mg. During each weighing, the filter must not be exposed for more
than a 2-min period to the laboratory atmosphere and relative humidity
above 50%. Alternatively, the filters may be oven-dried at 1058C (220°F)
for 2-3 h, desiccated for 2 h, and weighed.
7.2 Preliminary Field Determinations.
7.2.1 Select the sampling site and the minimum number of sampling
points according to EPA Method 1. Determine the stack pressure,
temperature, and range of velocity heads using EPA Method 2. It is
recommended that a leak-check of the pitot lines (see EPA Method 2,
Section 3.1) be performed. Determine the stack-gas moisture content using
EPA Method 4 or its alternatives to establish estimates of isokinetic
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sampling rate settings. Determine the stack gas dry molecular weight, as
described in EPA Method 2, Section 3.6. If integrated EPA Method 3
(Reference 1) sampling is used for molecular weight determination, the
integrated bag sample shall be taken simultaneously with, and for the same
total length of time as the sample run.
7.2.2 Select a nozzle size based on the range of velocity heads so
that it is not necessary to change the nozzle size to maintain isokinetic
sampling rates. During the run, do not change the nozzle. Ensure that
the proper differential pressure gauge is chosen for the range of velocity
heads encountered (see Section 2.2 of EPA Method 2).
7.2.3 Select a suitable probe liner and probe length so that all
traverse points can be sampled. For large stacks, to reduce the length of
the probe, consider sampling from opposite sides of the stack.
7.2.4 The total sampling time should be two hours. Allocate the same
time to all traverse points defined by EPA Method 1. To avoid timekeeping
errors, the length of time sampled at each traverse point should be an
integer or an integer plus one-half min. Size the condensate impinger for
the expected moisture catch or be prepared to empty it during the run.
7.3 Preparation of Sampling Train.
7.3.1 Add 50 ml of 0.05 M H2S04 to the condensate impinger, if used.
Place 100 ml of 0.05 M H2S04 in each of the next two impingers. Place 100
ml of 0.1 M NaOH in each of the following two impingers. Finally,
transfer approximately 200-300 g of preweighed silica gel from its
container to the last impinger. More silica gel may be used, but care
should be taken to ensure that it is not entrained and carried out from
the impinger during sampling. Place the silica gel container in a clean
place for later use in the sample recovery. Alternatively, the weight of
the silica gel plus impinger may be determined to the nearest 0.5 g and
recorded.
7.3.2 Using a tweezer or clean disposable surgical gloves, place a
labeled (identified) filter (weighed, if particulate matter is to be
determined) in the filter holder. Be sure that the filter is properly
centered and the gasket properly placed to prevent the sample gas stream
from circumventing the filter. Check the filter for tears after assembly
is completed.
7.3.3 To use glass liners, install the selected nozzle using a
Viton-A 0-ring when stack temperatures are <260°C (500°F) and a woven
glass fiber gasket when temperatures are higher. Other connecting systems
utilizing either 316 stainless steel or Teflon ferrules may be used. Mark
the probe with heat-resistant tape or by some other method to denote the
proper distance into the stack or duct for each sampling point.
7.3.4 Set up the train as in Figure 1. A minimal amount of silicone
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grease may be used on ground glass joints. Connect temperature sensors to
the appropriate potentiometer/display unit. Check all temperature sensors
at ambient temperature.
7.3.5 Place crushed ice around the impingers.
7.3.6 Turn on and set the filter and probe heating systems at the
desired operating temperatures. Allow time for the temperatures to
stabilize.
7.4 Leak-check Procedures.
7.4.1 Pretest leak-check. A pretest leak-check is recommended, but
not required. If the tester opts to conduct the pretest leak-check, the
following procedure shall be used.
7.4.1.1 If a Viton A 0-ring or other leak-free connection is
used in assembling the probe nozzle to the probe liner, leak-check
the train at the sampling site by plugging the nozzle and pulling a
380-mm Hg (15-in. Hg) vacuum.
NOTE: A lower vacuum may be used, provided that it is not exceeded during
the test.
7.4.1.2 If a woven glass fiber gasket is used, do not connect
the probe to the train during the leak-check. Instead, leak-check
the train by first plugging the inlet to the cyclone, if used, or
the filter holder and pulling a 380-mm Hg (15-in. Hg) vacuum (see
NOTE above). Then, connect the probe to the train and leak-check at
about 25-mm Hg (1-in. Hg) vacuum; alternatively, leak-check the
probe with the rest of the sampling train in one step at 380-mm Hg
(15-in. Hg) vacuum. Leakage rates in excess of 4% of the average
sampling rate or 0.00057 m /min (0.02 cfm), whichever is less, are
unacceptable.
7.4.1.3 The following leak-check instructions for the sampling
train may be helpful. Start the pump with bypass valve fully open
and coarse adjust valve completely closed. Partially open the
coarse adjust valve and slowly close the bypass valve until the
desired vacuum is reached. Do not reverse direction of the bypass
valve; this will cause water to back up into the filter holder. If
the desired vacuum is exceeded, either leak-check at this higher
vacuum or end the leak-check, as shown below, and start over.
7.4.1.4 When the leak-check is completed, first slowly remove
the plug from the inlet to the probe, cyclone, or filter holder and
immediately turn off the vacuum pump. This prevents the liquid in
the impingers from being forced backward into the filter holder and
silica gel from being entrained backward into the fifth impinger.
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7.4.2 Leak-checks during sample run. If, during the sampling run,
a component (e.g., filter assembly or impinger) change becomes necessary
or a port change is conducted, a leak-check shall be conducted immediately
after the interruption of sampling and before the change is made. The
leak-check shall be conducted according to the procedure outlined in
Section 7.4.1, except that it shall be conducted at a vacuum greater than
or equal to the maximum value recorded up to that point in the test. If
the leakage rate is found to be no greater than 0.00057 m3/min (0.02 cfm)
or 4% of the average sampling rate (whichever is less), the results are
acceptable. If a higher leakage rate is obtained, the tester shall void
the sampling run. Immediately after a component change or port change,
and before sampling is reinitiated, another leak-check similar to a pre-
test leak-check is recommended.
7.4.3 Post-test leak-check. A leak-check is mandatory at the
conclusion of each sampling run. The leak-check shall be done using the
same procedures as those with the pre-test leak-check, except that it
shall be conducted at a vacuum greater than or equal to the maximum value
reached during the sampling run. If the leakage rate is found to be no
greater than 0.00057 m /min (0.02 cfm) or 4% of the average sampling rate
(whichever is less), the results are acceptable. If a higher leakage rate
is obtained, the tester shall void the sampling run.
7.5 Train Operation.
7.5.1 During the sampling run, maintain an isokinetic sampling rate
to within 10% of true isokinetic. Maintain a temperature around the
filter and (cyclone, if used) of 120 ± 14°C (248 ± 25°F).
7.5.2 For each run, record the data required on a data sheet such
as the one shown in Figure 2. Be sure to record the initial dry gas meter
reading. Record the dry gas meter readings at the beginning and end of
each sampling time increment, when changes in flow rates are made before
and after each leak-check, and when sampling is halted. Take other
readings required by Figure 2 at least once at each sample point during
each time increment and additional readings when significant changes (20%
variation in velocity head readings) necessitate additional adjustments in
flow rate. Level and zero the manometer. Because the manometer level and
zero may drift due to vibrations and temperature changes, make periodic
checks during the traverse.
7.5.3 Clean the stack access ports prior to the test run to
eliminate the chance of sampling deposited material. To begin sampling,
remove the nozzle cap, verify that the filter and probe heating systems
are at the specified temperature, and verify that the pitot tube and probe
are positioned properly. Position the nozzle at the first traverse point,
with the tip pointing directly into the gas stream. Immediately start the
pump and adjust the flow to isokinetic conditions using a calculator or a
nomograph. Nomographs are designed for use when the Type S pitot tube
coefficient is 0.84 ± 0.02 and the stack gas equivalent density (dry
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molecular weight) is equal to 29 ± 4. If the stack gas molecular weight
and the pitot tube coefficient are outside the above ranges, do not use
the nomographs unless appropriate steps are taken to compensate for the
deviations (see Reference 3).
7.5.4 When the stack is under significant negative pressure
(equivalent to the height of the impinger stem), take care to close the
coarse adjust valve before inserting the probe into the stack, to prevent
water from backing into the filter holder. If necessary, the pump may be
turned on with the coarse adjust valve closed.
7.5.5 When the probe is in position, block off the openings around
the probe and stack access port to prevent unrepresentative dilution of
the gas stream.
7.5.6 Traverse the stack cross section, as required by EPA Method
1, being careful not to bump the probe nozzle into the stack walls when
sampling near the walls or when removing or inserting the probe through
the access port, in order to minimize the change of extracting deposited
material.
7.5.7 During the test run, make periodic adjustments to keep the
temperature around the filter holder (and cyclone, if used) at the proper
level. Add more ice, and, if necessary, salt to maintain a temperature of
<20°C (68°F) at the condenser/silica gel outlet.Also, periodically check
the level and zero of the manometer.
7.5.8 If the pressure drop across the filter becomes too high,
making isokinetic sampling difficult to maintain, it may be replaced in
the midst of a sample run. Using another complete filter holder assembly
is recommended, rather than attempting to change the filter itself. After
a new filter assembly is installed, conduct a leak-check. If determined,
the total particulate weight shall include the summation of all filter
assembly catches.
7.5.9 If the condensate impinger becomes too full, it may be
emptied, recharged with 50 ml of 0.05 M H2S04, and replaced during the
sample run. The condensate emptied must be saved and included in the
measurement of the volume of moisture collected and included in the sample
for analysis. The additional 50 ml of absorbing reagent must also be
considered in calculating the moisture. After the impinger is reinstalled
in the train, conduct a leak-check.
7.5.10 A single train shall be used for the entire sample run,
except in cases where simultaneous sampling is required in two or more
separate ducts or at two or more different locations within the same duct,
or in cases where equipment failure necessitates a change of trains.
7.5.11 Note that when two or more trains are used, separate analyses
of the particulate catch (if applicable) and the HC1 and C12 impinger
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catches from each train shall be performed, unless identical nozzle sizes
were used on all trains. In that case, the particulate catch and the HC1
and C12 impinger catches from the individual trains may be combined, and
a single particulate analysis and single HC1 and C12 analyses of the
impinger contents may be performed.
7.5.12 At the end of the sample run, turn off the coarse adjust
valve, remove the probe and nozzle from the stack, turn off the pump,
record the final dry gas meter reading.
7.5.13 If there is any possibility that liquid has collected in the
glass cyclone and/or on the filter, connect the Ascarite tube at the probe
inlet and operate the train with the filter heating system at 120 ± 14°C
(248 ± 25°F) at a low flow rate (e.g., AH = 1) sufficient to vaporize the
liquid and purge any HC1 in the cyclone or on the filter and pull it
through the train into the impingers. After 30 minutes, turn off the
flow, remove the Ascarite tube, and examine the cyclone and filter for any
visible moisture. If moisture is visible, repeat this step for 15
minutes.
7.5.14 Conduct a post-test leak-check. Also, leak-check the pitot
lines as described in EPA Method 2. The lines must pass this leak-check
in order to validate the velocity-head data.
7.5.15 If the moisture value is available, calculate percent
isokineticity (see Section 7.7.10) to determine whether the run was valid
or another test run should be conducted.
7.6 Sample Recovery
7.6.1 Allow the probe to cool. When the probe can be handled
safely, wipe off all the external surfaces of the tip of the probe nozzle
and place a cap over the tip. Do not cap the probe tip tightly while the
sampling train is cooling down because this will create a vacuum in the
filter holder, drawing water from the impingers into the holder.
7.6.2 Before moving the sampling train to the cleanup site, remove
the probe, wipe off any silicone grease, and cap the open outlet, being
careful not to lose any condensate that might be present. Wipe off any
silicone grease and cap the filter or cyclone inlet. Remove the umbilical
cord from the last impinger and cap the impinger. If a flexible line is
used between the first impinger and the filter holder, disconnect it at
the filter holder and let any condensed water drain into the first
impinger. Wipe off any silicone grease and cap the filter holder outlet
and the impinger inlet. Ground glass stoppers, plastic caps, serum caps,
Teflon tape, ParafilmR, or aluminum foil may be used to close these
openings.
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7.6.3 Transfer the probe and filter/impinger assembly to the cleanup
area. This area should be clean and protected from the weather to
minimize sample contamination or loss.
7.6.4 Save portions of all washing solutions used for cleanup
(acetone and reagent grade water) and the absorbing reagents (0.05 M H2S04
and 0.1 M NaOH) as blanks. Transfer 200 ml of each solution directly from
the wash bottle being used (rinse solutions) or the supply container
(absorbing reagents) and place each in a separate, prelabeled glass sample
container.
7.6.5 Inspect the train prior to and during disassembly and note any
abnormal conditions.
7.6.6 Container No. 1 (filter catch for particulate determination).
Carefully remove the filter from the filter holder and place it in its
identified Petri dish container. Use one or more pair of tweezers to
handle the filter. If it is necessary to fold the filter, ensure that the
particulate cake is inside the fold. Carefully transfer to the Petri dish
any particulate matter or filter fibers that adhere to the filter holder
gasket, using a dry nylon bristle brush or sharp-edged blade, or both.
Label the container and seal with Teflon tape around the circumference of
the lid.
7.6.7 Container No. 2 (front-half rinse for particulate
determination). Taking care that dust on the outside of the probe or
other exterior surfaces does not get into the sample, quantitatively
recover particulate matter or any condensate from the probe nozzle, probe
fitting, probe liner, and front half of the filter holder by washing these
components with acetone into a glass container. Retain an acetone blank
and analyze with the samples.
7.6.8 Perform rinses as follows: carefully remove the probe nozzle
and clean the inside surface by rinsing with acetone from a wash bottle
and brushing with a nylon bristle brush. Brush until the rinse shows no
visible particles; then make a final rinse of the inside surface with the
acetone. Brush and rinse the inside parts of the Swagelok fitting with
the acetone in a similar way until no visible particles remain.
7.6.9 Have two people rinse the probe liner with acetone by tilting
and rotating the probe while squirting acetone into its upper end so that
all inside surfaces will be wetted with solvent. Let the acetone drain
from the lower end into the sample container. A glass funnel may be used
to aid in transferring liquid washed to the container.
7.6.10 Follow the acetone rinse with a probe brush. Hold the probe
in an inclined position and squirt acetone into the upper end while
pushing the probe brush through the probe with a twisting action; place a
sample container underneath the lower end of the probe and catch any
acetone and particulate matter that is brushed from the probe. Run the
0050 - 13 Revision 0
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brush through the probe three or more times until no visible particulate
matter is carried out with the acetone or none remains in the probe liner
on visual inspection. Rinse the brush with acetone and quantitatively
collect these washings in the sample container. After the brushing, make
a final acetone rinse of the probe as described above. Between sampling
runs, keep brushes clean and protected from contamination.
7.6.11 Clean the inside of the front half of the filter holder and
cyclone by rubbing the surfaces with a nylon bristle brush and rinsing
with acetone. Rinse each surface three times, or more if needed, to
remove visible particulate. Make a final rinse of the brush and filter
holder. Carefully rinse out the glass cyclone and cyclone flask (if
applicable). Brush and rinse any particulate material adhering to the
inner surfaces of these components into the front-half rinse sample.
After all rinses and particulate matter have been collected in the sample
container, tighten the lid on the sample container so that acetone will
not leak out when it is shipped to the laboratory. Mark the height of the
fluid level to determine whether leakage occurs during transport. Label
the container to identify its contents.
7.6.12 Container No. 3 (knockout and acid impinger catch for
moisture and HC1 determination). Disconnect the impingers. Measure the
liquid in the acid and knockout impingers to within ± 1 mL by using a
graduated cylinder or by weighing it to within ± 0.5 g by using a balance
(if one is available). Record the volume or weight of liquid present.
This information is required to calculate the moisture content of the
effluent gas. Quantitatively transfer this liquid to a leak-free sample
storage container. Rinse these impingers and the connecting glassware
(and tubing, if used) with water, and add these rinses to the storage
container. Seal the container, shake to mix, and label. The fluid level
should be marked so that if any sample is lost during transport, a
correction proportional to the lost volume can be applied.Retain rinse
water and acidic absorbing solution blanks and analyze with the samples.
7.6.13 Container No. 4 (alkaline impinger catch for C12 and moisture
determination). Measure and record the liquid in the alkaline impingers
as described in Section 7.6.12. Quantitatively transfer this liquid to a
leak-free sample storage container. Rinse these two impingers and
connecting glassware with water and add these rinses to the container.
Seal the container, shake to mix, and label; mark the fluid level. Retain
alkaline absorbing solution blank and analyze with the samples.
7.6.14 Container No. 5 (silica gel for moisture determination).
Note the color of the indicating silica gel to determine if it has been
completely spent and make a notation of its condition. Transfer the
silica gel from the last impinger to its original container and seal. A
funnel may make it easier to pour the silica gel without spilling. A
rubber policeman may be used as an aid in removing the silica gel from the
impinger. It is not necessary to remove the small amount of dust
particles that may adhere strongly to the impinger wall. Because the gain
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in weight is to be used for moisture calculations, do not use any water or
other liquids to transfer the silica gel. If a balance is available in
the field, weigh the container and its contents to 0.5 g or better.
7.6.15 Prior to shipment, recheck all sample containers to ensure
that the caps are well secured. Seal the lids of all containers around the
circumference with Teflon tape. Ship all liquid samples upright and all
particulate filters with the particulate catch facing upward.
7.7 Calculations. Retain at least one extra decimal figure beyond those
contained in the available data in intermediate calculations, and round off only
the final answer appropriately.
7.7.1 Nomenclature.
An = Cross-sectional area of nozzle, m2 (ft2).
Bws = Water vapor in the gas stream, proportion by volume.
Ca = Acetone blank residue concentration, mg/mg.
Cd = Type S pitot tube coefficient (nominally 0.84 ± 0.02),
dimensionless.
cs = Concentration of particulate matter in stack gas, dry basis,
corrected to standard conditions, g/dscm (g/dscf).
I = Percent of isokinetic sampling.
ma = Mass of residue of acetone after evaporation, mg.
Mn = Total amount of particulate matter collected, mg.
Md = Stack-gas dry molecular weight, g/g-mole (Ib/lb-mole).
Mw = Molecular weight of water, 18.0 g/g-mole (18.0 Ib/lb-
mole).
Pbar = Barometric pressure at the sampling site, mm Hg (in. Hg).
Ps = Absolute stack-gas pressure, mm Hg (in. Hg).
Pstd = Standard absolute pressure, 760 mm Hg (29.92 in. Hg).
R = Ideal gas constant, 0.06236 mm Hg-m3 (K-g-mole (21.85 in. Hg-
ft3/°R-lb-mole).
Tm = Absolute average dry-gas meter temperature (see Figure
2), °K, (°R).
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Ts = Absolute average stack-gas temperature (see Figure 2), °K
(°R).
Tstd = Standard absolute temperature, 293°K (528°R).
Vlc = Total volume of liquid collected in the impingers and silica
gel, ml.
Vm = Volume of gas sample as measured by dry-gas meter, dscm
(dscf).
= Volume of gas sample measured by the dry-gas meter, corrected
to standard conditions, dscm (dscf).
Vwistd) = Volume of water vapor in the gas sample, corrected to standard
conditions, scm (scf).
Vs = Stack-gas velocity, calculated by Method 2, Equation 2-9,
using data obtained from Method 5, m/sec (ft/sec).
Wa = Weight of residue in acetone wash, mg.
Va = Volume of acetone blank, mL.
Vaw = Volume of acetone used in wash; ml.
Y = Dry-gas-meter calibration factor, dimensionless.
H = Average pressure differential across the orifice meter, mm H20
(in. H20).
a = Density of acetone, mg/jj] (see label on bottle).
w = Density of water, 0.9982 g/mL (0.002201 Ib/mL).
0 = Total sampling time, min.
13.6 = Specific gravity of mercury.
60 = Sec/min.
100 = Conversion to percent.
7.7.2 Average dry gas meter temperature and average orifice pressure
drop. See data sheet (Figure 2).
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7.7.3 Dry gas volume. Correct the sample measured by the dry gas
meter to standard conditions (20°C, 760 mm Hg [68°F, 29.92 in. Hg]) by
using Equation 1:
Fn T y _ uY Tstd Pbar + AH/13.6 _ Pbar+ H/13.6
tq. 1 Vm(stdl - VmY ~ N VmY
Pstd Tn
where: KT = 0.3858 K/mm Hg for metric units, or
K, = 17.64°R/in. Hg for English units.
7.7.4 Volume of water vapor.
n RT
Eq- 2 Vw(8tdl = Vlc __Pw_Jl!:td = K2 Vlc
Mw Pstd
where: K2 = 0.001333 m3/ml for metric units, or
K2 = 0.04707 m3/mi for English units.
7.7.5 Moisture content.
Eq. 3 Bws = V«
V + V
•mlstdl T "wlstdl
NOTE: In saturated or water-droplet-laden gas streams, two calculations of
the moisture content of the stack gas shall be made, one from the impinger
analysis (Equation 3) and a second from the assumption of saturated
conditions. The lower of the two values of Bw shall be considered
correct. The procedure for determining the moisture content based upon
assumption of saturated conditions is given in the Note to Section 1.2 of
Method 4. For the purposes of this method, the average stack gas
temperature from Figure 2 may be used to make this determination, provided
that the accuracy of the in-stack temperature sensor is ± 1°C (2°F).
7.7.6 Acetone blank concentration. For particulate determination.
r n r "1.
Eq. 4 Ca = a
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7.7.7 Acetone wash blank. For participate determination.
Wa = Ca Vaw pa
7.7.8 Total particulate weight. Determine the total participate
catch from the sum of the weights obtained from Container Nos. 1 and 2
less the acetone blank (WJ .
7.7.9 Particulate concentration.
cs = (0.001 g/mg)(myvm(std))
7.7.10 Isokinetic variation.
7.4.10.1 Calculation from raw data.
= 10° TstK3Flc + (VJTJ (Pbar + H/13.6)]
Eq 8
600VsPsAn
where: K3 = 0.003454 mm Hg-m3/mL-K for metric units, or
K3 = 0.002669 in. Hg-ft3/mL °R for English units.
7.4.10.2 Calculation for intermediate values.
T = TsVm(stdlPstd100
TstdVs0AnPs60(l-Bws)
T V
's'mlstd)
PsVsAn 0 (1-BWS)
where: K4 = 4.320 for metric units, or
K4 = 0.09450 for English units.
7.7.11 Acceptable results. If 90% < I < 110%, the results are
acceptable.
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7.7.12 Analytical calculation for total /vg HC1 per sample.
Calculate as described below:
mHCl = (S-B) x Vs x 36.46/35.453 (1)
where: mHCl = Mass of HC1 in sample, ug,
S = Analysis of sample, us Cl'/mL,
B = Analysis of reagent blank, ug Cl'/mL,
Vs = Volume of filtered and diluted sample, mL,
36.46 = Molecular weight of HC1, ug/ug-mole, and
35.45 = Atomic weight of CT, ug/ug-mole.
7.4.13 Analytical calculation for total ug C12 per sample.
Calculate as described below:
MC12 = (S-B) x Vs x 70.91/35.45 (2)
where: Mcl2 = Mass of C12 in sample, ug,
70.91 = Molecular weight of C12, ug/ug-mole, and
35.45 = Atomic weight of Cl", ug/ug-mole.
7.4.14 Concentration of HC1 in the flue gas. Calculate as
described below:
C = K x m/Vm(std) (3)
where: C = Concentration of HC1 or C12, dry basis, mg/dscm,
K = 10"3 mg/ug,
m = Mass of HC1 or C12 in sample, ug, and
Vm(std) = Dry gas volume measured by the dry gas meter,
corrected to standard conditions, dscm.
8.0 QUALITY CONTROL
8.1 Sampling. See EPA Manual 600/4-77-027b for Method 5 quality control.
8.2 Analysis. At the present time, a validated audit material does not
exist for this method. Analytical quality control procedures are detailed in
Method 9057.
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8.3 Quality control check sample. Chloride solutions of reliably known
concentrations are available for purchase from the National Institute of
Standards and Technology (SRM 3182). The QC check sample should be prepared in
the appropriate absorbing reagent at a concentration approximately equal to the
mid range calibration standard. The quality control check sample should be
injected in duplicate immediately after the calibration standards have been
injected the first time. The Cl" value obtained for the check sample using the
final calibration curve should be within 10 percent of the known value for the
check sample.
9.0 METHOD PERFORMANCE
9.1 The in-stack detection limit for the method is approximately 0.02 ug
of HC1 per liter of stack gas. The method has a negative bias below 20 ppm HC1
(Reference 6).
9.2 It is preferable to include the cyclone in the sampling train to
protect the filter from any moisture present. There is research in progress
regarding the necessity of the cyclone at low moisture sources and the use of
Ascarite II in the drying procedure (Section 7.5.12).
9.3 The lower detection limit of the analytical method is 0.1 ug of Cl"
per ml of sample solution. Samples with concentrations which exceed the linear
range of the 1C may be diluted.
9.4 The precision and bias for analysis of HC1 using this analytical
protocol have been measured in combination with the midget impinger HC1/C12 train
(Method 0051) for sample collection. The laboratory relative standard deviation
is within 6.2 percent and 3.2 percent at HC1 concentrations of 3.9 and 15.3 ppm.
respectively. The method does not exhibit any bias for HC1 when sampling at C12
concentrations less than 50 ppm.
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10.0 REFERENCES
1. U. S. Environmental Protection Agency, 40 CFR Part 60, Appendix A, Methods
1-5.
2. U. S. Environmental Protection Agency, "Quality Assurance Handbook for Air
Pollution Measurement Systems, Volume III, Stationary Source Specific
Methods," Publication No. EPA-600/4-77-027b, August 1977.
3. Shigehara, R. T., Adjustments in the EPA Nomography for Different Pitot
Tube Coefficients and Dry Molecular Weights, Stack Sampling News, £:4-ll
(October 1974).
4. Steinsberger, S. C. and J. H. Margeson, "Laboratory and Field Evaluation
of a Methodology for Determination of Hydrogen Chloride Emissions from
Municipal and Hazardous Waste Incinerators," U. S. Environmental
Protection Agency, Office of Research and Development, Report No.
600/3/89/064, April, 1989. Available from NTIS.
5. State of California, Air Resources Board, Method 421, "Determination of
Hydrochloric Acid Emissions from Stationary Sources," March 18, 1987.
6. Entropy Environmentalists, Inc., "Laboratory Evaluation of a Sampling and
Analysis Method for Hydrogen Chloride Emissions from Stationary Sources:
Interim Report," EPA Contract No. 68-02-4442, Research Triangle Park,
North Carolina, January 22, 1988.
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Figure 1. Isokinetic HC1/C1? sampling train.
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METHOD 0050
ISOKINETIC HC1/C1, EMISSION SAMPLING TRAIN
7 1.1 - 7.1,4 Preliminary Procedures:
Maintain and calibrate all sampling
equipment, prepare reagents and
filters (if required}.
7.2.1 Make preliminary measurements
to establish estimates of isokmetic
sampling rates and sites.
7 2 2 - 7 2.3 Select appropriate
nozzel size to maintain isokinetic
sampling rate; select apprpnate
probe liner and probe lengths.
7 3.1 • 7 3.2 Prepare sampling train,
add reagents to impingers. Place
preweighed filter in filter holder if
particulate matter is to be measured.
7.3.3 - 7 3.4 Select appropriate
nozzles and fiber gasket; set-up
sampling train, check temp.
settings at ambient temp.
741-7 4.3 Perform leak-check prior
to sampling run and during run if
any component is changed.
0050 - 24
7.5 1 - 7.5.11 Verify filter and probe
heating temps , maintain an isokmetic
sampling rate within 10% true
isokmatic, position nozzel directly in
gas stream, initiate sampling.
7.5.12 At the end of sampling run,
remove nozzel, turn off pump.
7 5.13 Recover any liquid m
cyclone and/or filter by attaching
an Ascante tube to probe inlet
and operate tram 1 20 + /- 1 4° C for
30 mm.
7.5.14 - 7 5.15 Perform post-teat leak-
check, if moisture valve is known,
calculate % isokmeticity.
i
7.6.1 - 7.6.3 Allow probe to cool,
disassemble umbilical cord, cap,
and transfer to clean-up area
764 Collect wash solutions and
absorbing reagents for field
blank analysts.
7 6.6 Remove filter from filter
holder (if required), place in
petn dish, label as container
No. 1
7.6.7 - 7.6.11 Quantitatively recover
particulate matter from half of probe
assembly and filter holder. Combine
nnses and label as container No. 2.
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METHOD 0050 (Cont.)
ISOKINETIC HC1/C1, EMISSION SAMPLING TRAIN
7.6.12 - 7.6.1 3 Disconnect knockout
fmpinger and 0.1N H SO impingers
(impinger 2 and 3). Record weight or
volume of solutions, rinse assembly,
combine solutions and rinses into
container No. 3. Repeat procedure
for alkaline impinger solutions, label as
container No.4.
7.6.14 Weigh silica gel from last
impinger, transfer to original
container.
Analyze impinger solutions for total
Cl using Method 9056
(Ion Chromatography).
7.7.1 - 7.7.14 Calculate total particulate
concentration (7.7.9) and total HCL
concentration (7.7.14) in flue as.
I
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METHOD 0051
MIDGET IMPINGER HC1/C1, EMISSION SAMPLING TRAIN
1.0 SCOPE AND APPLICATION
1.1 This method describes the collection of hydrogen chloride (HC1, CAS
Registry Number 7647-01-0) and chlorine (C12, CAS Registry Number 7782-50-5) in
stack gas emission samples from hazardous waste incinerators and municipal waste
combustors. The collected samples are analyzed using Method 9057. This method
is designed to collect HC1/C12 in their gaseous forms. Use of this method is
limited to the sampling of relatively dry, particulate-free gas streams.
Sources, such as those controlled by wet scrubbers, that emit acid particulate
matter (e.g., HC1 dissolved in water droplets) must be sampled using an
isokinetic HC1/C12 sampling train (see Method 0050).
2.0 SUMMARY OF METHOD
2.1 An integrated gas sample is extracted from the stack and passes
through a particulate filter, acidified water, and finally through an alkaline
solution. The filter serves to remove particulate matter such as chloride salts
which could potentially react and form the analyte in the absorbing solutions.
In the acidified water absorbing solution, the HC1 gas is solubilized and forms
chloride ions (CV) as follows:
HC1 + H20 = H304 + CV
The C12 gas present in the emissions has a very low solubility in acidified water
and passes through to the alkaline absorbing solution where it undergoes
hydrolysis to form a proton (H+), CT, and hypochlorous acid (HC10) as follows:
H20 + C12 = H+ + CT + HC10
The Cl" ions in the separate solutions are measured by ion chromatography (Method
9057).
3.0 INTERFERENCES
3.1 Volatile materials which produce chloride ions upon dissolution
during sampling are obvious interferences in the measurement of HC1. One
interferant for HC1 is diatomic chlorine (C12) gas which disproportionates to HC1
and hypochlorous acid (HC10) upon dissolution in water. C12 gas exhibits a low
solubility in water, however, and the use of acidic rather than neutral or basic
solutions for collection of hydrogen chloride gas greatly reduces the dissolution
of any chlorine present. Sampling a 400 ppm HC1 gas stream containing 50 ppm C12
with this method does not cause a significant bias. Sampling a 200 ppm HC1 gas
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stream containing 180 pptn C12 results in a positive bias of 3.4 percent in the
HC1 measurement.
4.0 APPARATUS AND MATERIALS
4.1 Sampling Train. The sampling train is shown in Figure 1 and
component parts are discussed below.
4.1.1 Probe. Borosilicate glass, approximately 3/8-in. (9-mm)
inside diameter, with a heating system to prevent condensation. When the
concentration of alkaline particulate matter in the emissions is high, a
3/8-in. (9-mm) inside diameter Teflon elbow should be attached to the
inlet of the probe. A 1-in. (25-mm) length of Teflon tubing with a 3/8-
in. (9-mm) inside diameter should be attached at the open end of the elbow
to permit the opening of the probe to be turned away from the gas stream,
thus reducing the amount of particulate entering the train. When high
concentrations of particulate matter are not present, the Teflon elbow is
unnecessary, and the probe inlet can be perpendicular to the gas stream.
When sampling at locations where gas temperatures are greater than
approximately 400°F, such as wet scrubber inlets, glass or quartz elbows
must be used. In no case should a glass wool plug be used to remove
particulate matter; use of such a filtering device could result in a bias
in the data. Instead, a Teflon filter should be used as specified in
Section 5.5.
4.1.2 Three-way stopcock. A borosilicate, three-way glass stopcock
with a heating system to prevent condensation. The heated stopcock should
connect directly to the outlet of the probe and filter assembly and the
inlet of the first impinger. The heating system should be capable of
preventing condensation up to the inlet of the first impinger. Silicone
grease may be used, if necessary, to prevent leakage.
4.1.3 Impingers. Five 30-ml midget impingers with leak-free glass
connectors. Silicone grease may be used, if necessary, to prevent
leakage. For sampling at high moisture sources or for extended sampling
times greater than one hour, a midget impinger with a shortened stem (such
that the gas sample does not bubble through the collected condensate)
should be used in front of the first impinger.
4.1.4 Mae West impinger or drying tube. Mae West design impinger
(or drying tube, if a moisture determination is not to be conducted)
filled with silica gel, or equivalent, to dry the gas sample and to
protect the dry gas meter and pump.
4.1.5 Sample line. Leak-free, with compatible fittings to connect
the last impinger to the needle valve.
4.1.6 Barometer. Mercury, aneroid, or other barometer capable of
measuring atmospheric pressure within 2.5 mm Hg (0.1 in Hg). In many
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cases, the barometric reading may be obtained from a nearby National
Weather Service station, in which case the station value (which is the
absolute barometric pressure) shall be requested and an adjustment for the
elevation differences between the weather station and sampling point shall
be applied at a rate of minus 2.5 mm Hg (0.1 in. Hg) per 30 m (100 ft)
elevation increase or vice versa for elevation decrease.
4.1.7 Purge pump, purge line, drying tube, needle valve, and rate
meter. Pump capable of purging sample probe at 2 liters/min. with drying
tube, filled with silica gel or equivalent, to protect pump, and a rate
meter, 0 to 5 liters/min.
4.1.8 Metering system. The following items comprise the metering
system which is identical to that used for EPA Method 6 (see Reference 5).
4.1.8.1 Valve. Needle valve, to regulate sample gas flow
rate.
4.1.8.2 Pump. Leak-free diaphragm pump, or equivalent, to
pull gas through train. Install a small surge tank between the pump
and the rate meter to eliminate the pulsation effect of the
diaphragm pump on the rotameter.
4.1.8.3 Rate meter. Rotameter, or equivalent, capable of
measuring flow rate to within 2 percent of selected flow rate of 2
liters/min.
4.1.8.4 Volume meter. Dry gas meter, sufficiently accurate
to measure the sample volume within 2 percent, calibrated at the
selected flow rate and conditions encountered during sampling, and
equipped with a temperature gauge (dial thermometer^or equivalent)
capable of measuring temperature to within 3°C (5.4°F).
4.1.8.5 Vacuum gauge. At least 760 mm Hg (30 in. Hg) gauge
to be used for leak check of the sampling train.
4.1.9 Water Bath: To minimize loss of absorbing solution
4.2 Sample Recovery.
4.2.1 Wash bottles. Polyethylene or glass, 500 ml or larger, two.
4.2.2 Storage bottles. Glass, with Teflon-lined lids, 100 ml, to
store impinger samples (two per sampling run).
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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 reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Reagent Water. All references to water in the method refer to
reagent water as defined in Chapter One unless otherwise specified. It is
advisable to analyze a blank sample of this reagent prior to sampling, since the
reagent blank value obtained during the field sample analysis must be less than
10 percent of the sample values.
5.3 Sulfuric acid (0.05 M), H2SO,. Used as the HC1 absorbing reagent.
To prepare 100 ml, slowly add 0.28 ml of concentrated H,S04 to about 90 ml of
water while stirring, and adjust the final volume to IOQ ml using additional
water. Shake well to mix the solution. It is advisable to analyze a blank
sample of this reagent prior to sampling, since the reagent blank value obtained
during the field sample analysis must be less than 10 percent of the sample
values.
5.4 Sodium hydroxide (0.1 M), NaOH. Used as the C12 absorbing reagent.
To prepare 100 ml, dissolve 0.40 g of solid NaOH in about 90 ml of water and
adjust the final volume to 100 ml using additional water. Shake well to mix the
solution. It is advisable to analyze a blank sample of this reagent prior to
sampling, since the reagent blank value obtained during the field sample analysis
must be less than 10 percent of the sample value.
5.5 Filter. Teflon mat Pallflex® TX40HI75 or equivalent. Locate in a
glass, quartz, or Teflon filter holder with a Teflon filter support in a filter
box heated to 250°F.
5.6 Stopcock grease. Acetone-insoluble, heat-stable silicone grease may
be used, if necessary.
5.7 Silica gel. Indicating type, 6- to 16-mesh. If the silica gel has
been used previously, dry at 175°C (350°F) for 2 hours. New silica gel may be
used as received. Alternatively, other types of desiccants (equivalent or
better) may be used.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Sample collection is described in this method. The analytical
procedures are described in Method 9057.
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6.2 Samples should be stored in clearly labeled, tightly sealed
containers between sample recovery and analysis. They may be analyzed up to four
weeks after collection.
7.0 PROCEDURE
7.1 Calibration. Section 3,5.2 of EPA's Quality Assurance Handbook
Volume III (Reference 4) may be used as a guide for these operations.
7.1.1 Dry Gas Metering System.
7.1.1.1 Initial calibration. Before its initial use in the
field, first leak check the metering system (sample line, drying
tube, if used, vacuum gauge, needle valve, pump, rate meter, and dry
gas meter) as follows: plug the inlet end of the sampling line,
pull a vacuum of 250 mm (10 in.) Hg, plug off the outlet of the dry
gas meter, and turn off the pump. The vacuum should remain stable
for 30 seconds. Carefully release the vacuum from the system by
slowly removing the plug from the sample line inlet. Remove the
sampling line (and drying tube, if applicable), and connect the dry
gas metering system to a appropriately sized wet test meter (e.g.,
1 liter per revolution). Make three independent calibration runs,
using at least five revolutions of the dry gas meter per run.
Calculate the calibration factor, Y (wet test meter calibration
volume divided by the dry gas meter volume, with both volumes
adjusted to the same reference temperature and pressure), for each
run, and average the results. If any Y value deviates by more than
2 percent from the average, the metering system is unacceptable for
use. Otherwise, use the average as the calibration factor for
subsequent test runs.
7.1.1.2 Post-test calibration check. After each field test
series, conduct a calibration check as in Section 7.1.1.1 above,
except for the following variations: (a) the leak check is not to
be conducted, (b) three or more revolutions of the dry gas meter may
be used, (c) only two independent runs need to be made. If the
calibration factor does no deviate by more than 5 percent from the
initial calibration factor (determined in Section 7.1.1.1), the dry
gas meter volumes obtained during the test series are acceptable.
If the calibration factor deviates by more than 5 percent,
recalibrate the metering system as Section 7.1.1.1, and for the
calculations, use the calibration factor (initial or recalibration)
that yields the lower gas volume for each test run.
7.1.2 Thermometer(s). Prior to each field test, calibrate against
mercury-in-glass thermometers at ambient temperature. If the thermometer
being calibrated reads within 2°C (2.6°F) of the mercury-in-glass
thermometer, it is acceptable. If not, adjust the thermometer or use an
appropriate correction factor.
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7.1.3 Rate meter. The rate meter should be cleaned and maintained
according to the manufacturer's instructions.
7.1.4 Barometer. Prior to each field test, calibrate against a
mercury barometer. The field barometer should agree within 0.1 in. Hg
with the mercury barometer. If it does not, the field barometer should be
adjusted.
7.2 Sampling.
7.2.1 Preparation of collection train. Prepare the sampling train
as follows: The first or knockout impinger should have a shortened stem
and be left empty to condense moisture in the gas stream. The next two
midget impingers should each be filled with 15 ml of 0.05 M H2SO,. The
fourth and fifth impingers should each be filled with 15 ml of 0.1 M NaOH.
Place a fresh charge of silica gel, or equivalent, in the Mae West
impinger (or the drying tube). Connect the impingers in series with the
knockout impinger first, followed by the two impingers containing the
acidified reagent and two impingers containing the alkaline reagent, and
the Mae West impinger containing the silica gel. If the moisture will be
determined, weigh the impinger assembly to the nearest + 0.5 g and record
the weight.
7.2.2 Leak check procedures. Leak check the probe and three-way
stopcock prior to inserting the probe into the stack. Connect the
stopcock to the outlet of the probe, and connect the sample line to the
needle valve. Plug the probe inlet, turn on the sample pump, and pull a
vacuum of at least 250 mm Hg (10 in. Hg). Turn off the needle valve, and
note the vacuum gauge reading. The vacuum should remain stable for at
least 30 seconds. Place the probe in the stack at the sampling location,
and adjust the filter heating system at 250°F and the probe and stopcock
heating systems to a temperature sufficient to prevent water condensation.
Connect the first impinger to the stopcock, and connect the sample line to
the last impinger and the needle valve. Upon completion of a sampling
run, remove the probe from the stack and leak check as described above.
If a leak has occurred, the sampling run must be voided. Alternatively,
the portion of the train behind the probe may be leak checked between
multiple runs at the same site as follows: Close the stopcock to the
first impinger, and turn on the sample pump. Pull a vacuum of at least
250 mm Hg (10 in. Hg), turn off the needle valve, and note the vacuum
gauge reading. The vacuum should remain stable for at least 30 seconds.
Release the vacuum on the impinger train by turning the stopcock to the
vent position to permit ambient air to enter. If this procedure is used,
the full train leak check described above must be conducted following the
final run and all preceding sampling runs voided if a leak has occurred.
7.2.3 Purge procedures. Immediately prior to sampling, connect the
purge line to the stopcock and turn the stopcock to permit the purge pump
to purge the probe (see Figure 1A). Turn on the purge pump, and adjust
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the purge rate to 2 liters/min. Purge for at least 5 minutes prior to
sampling.
7.2.4 Sample collection. Turn on sample pump, pull a slight vacuum
of approximately 25 mm Hg (1 in. Hg) on the impinger train, and turn the
stopcock to permit stack gas to be pulled through the impinger train (see
Figure 1C). Adjust the sampling rate to 2 liters/min. as indicated by the
rate meter, and maintain this rate within 10 percent during the entire
sampling run. Take readings of the dry gas meter, the dry gas meter
temperature, rate meter, and vacuum gauge at least once every five minutes
during the run. A sampling time of one hour is recommended. However, if
the expected condensate catch for this sampling run duration will exceed
the capacity of the sampling train, (1) a larger knockout impinger may be
used or (2) two sequential half-hour runs may be conducted. At the
conclusion of the sampling run, remove the train from the stack, cool, and
perform a leak check as described in Section 7.2.2.
7.3 Sample recovery. Following sampling, disconnect the impinger train
from the remaining sampling equipment at the inlet to the knockout impinger and
the outlet to the last impinger. If performing a moisture determination, wipe
off any moisture on the outside of the train and any excess silicone grease at
the inlet and outlet openings; weigh the train to the nearest 0.5 g and record
this weight. Then disconnect the impingers from each other. Quantitatively
transfer the contents of the first three impingers (the knockout impinger and the
two 0.05 M HpS04 impingers) to a leak-free storage bottle. Add the water rinses
of each of tnese impingers and connecting glassware to the storage bottle. The
contents of the impingers and connecting glassware from the second set of
impingers (containing the 0.1 M NaOH) should be recovered in a similar manner if
a Clp analysis is desired. The sample bottle should be sealed, shaken to mix,
and labeled; the fluid level should be marked so that if any sample is lost
during transport, a correction proportional to the lost volume can be applied.
Save portions of the 0.05 M H2S04 and 0.1 M NaOH used as impinger reagents as
reagent blanks. Take 50 ml of each and place in separate leak-free storage
bottles. Label and mark the fluid levels as previously described.
7.4 Calculations. Retain at least one extra decimal figure beyond those
contained in the available data in intermediate calculations, and round off only
the final answer appropriately.
7.4.1 Nomenclature.
Bws = Water vapor in the gas stream, proportion by
volume.
M = Molecular weight of water, 18.0 g/g-mole
(18.0 Ib/lb-mole).
Pbar = Barometric pressure at the exit orifice of the
dry gas meter, mm Hg (in. Hg).
Pstd = Standard absolute pressure, 760 mm Hg
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P , = Standard absolute pressure, 760 mm Hg
(29.92 in. Hg).
R = Ideal gas constant, 0.06236 mm Hg-m3/°K-g-mole
(21.85 in. Hg-ft3/°R-lb-mo1e).
Tm = Average dry gas meter absolute temperature,
°K (°R).
Tstd = Standard absolute temperature, 293°K (528°R).
Vtc = Total volume of liquid collected in impingers
and silica gel, ml (equivalent to the
difference in weight of the impinger train
before and after sampling, 1 mg = 1 ml).
Vm = Dry gas volume as measured by the dry gas
meter, dcm (dcf).
Vm(std) = Dry gas volume measured by the dry gas meter,
corrected to standard conditions, dscm (dscf).
VH(std) = Volume of water vapor in the gas sample,
corrected to standard conditions, scm (scf).
Y = Dry gas meter calibration factor.
pa = Density of water, 0.9982 g/mL (0.002201 Ib/mL).
7.4.2 Sample volume, dry basis, corrected to standard conditions.
Calculate as described below:
T P UP
'std rbar vm rbar
u = V Y = K Y
Vstd) Vm jm p T
where:
= 0.3858°K/mm Hg for metric units.
= 17.64°R/in. Hg for English units.
7.4.3 Volume of water vapor.
v -V --.- = KV (2)
Vstd) Vlc "Vic ^1
Mw Pstd
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where:
K2 = 0.0013333 m3/ml_ for metric units.
= 0.04707 ft3/mL for English units.
7.4.4 Moisture content.
Bus *«!»---- (3)
V + V
Ym(std) T 'w(std)
7.4.4 Analytical calculation of total ug HC1 per sample. Calculate
as described below:
mHcl = (S-B) x Vs x 36.46/35.45 (1)
where: mHcl = Mass of HC1 in sample, ug,
S = Analysis of sample, ug CV/mL,
B = Analysis of reagent blank, ug CV/mL,
Vs = Volume of filtered and diluted sample, ml,
36.46 = Molecular weight of HC1, ug/ug-mole, and
35.45 = Atomic weight of Cl", ug/ug-mole.
7.4.5 Analytical calculation of total ug C12 per sample. Calculate
as described below:
MC12 = (S-B) x Vs x 70.91/35.45 (2)
where: MC12 = Mass of C12 in sample, ug,
70.91 = Molecular weight of C12, ug/ug-mole, and
35.45 = Atomic weight of Cl", ug/ug-mole.
S = Analysis of sample, ug CV/mL,
B = Analysis of reagent blank, ug CV/mL,
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below:
7.4.6 Concentration of HC1 in the flue gas. Calculate as described
c = K x m/Vm(std) (3)
where: C = Concentration of HC1 or C12, dry basis, mg/dscm,
K = 10"3 mg/ug,
m = Mass of HC1 or C12 in sample, ug, and
Vm(std) = Dry gas volume measured by the dry gas meter,
corrected to standard conditions, dscm.
8.0 QUALITY CONTROL
8.1 At the present time, a validated audit material does not exist for
this method. Analytical quality control procedures are detailed in Method 9057.
8.2 Quality control check sample. Chloride solutions of reliably known
concentrations are available for purchase from the National Bureau of Standards
(SRM 3182). The QC check sample should be prepared in the appropriate absorbing
reagent at a concentration approximately equal to the mid range calibration
standard. The quality control check sample should be injected in duplicate
immediately after the calibration standards have been injected the first time.
The Cl" value obtained for the check sample using the final calibration curve
should be within 10 percent of the known value for the check sample.
9.0 METHOD PERFORMANCE
9.1 The in-stack detection limit for the method is approximately 0.08 ug
of HC1 per liter of stack gas for a 1-hour sample.
9.2 The precision and bias for measurement of HC1 using this sampling
protocol combined with the analytical protocol for Method 0050 have been
determined. The laboratory relative standard deviation is within 6.2 percent and
3.2 percent at HC1 concentrations of 3.9 and 15.3 ppm, respectively. The method
does not exhibit
50 ppm.
any bias for HC1 when sampling at C12 concentrations less than
10.0 REFERENCES
Steinsberger, S.C. and J.H. Margeson, "Laboratory and Field Evaluation of
a Methodology for Determination of Hydrogen Chloride Emissions from
Municipal and Hazardous Waste Incinerators," U.S. Environmental Protection
Agency, Office of Research and Development, Report No. 600/3/89/064,
April, 1989. Available from NTIS.
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2. State of California, Air Resources Board, Method 421, "Determination of
Hydrochloric Acid Emissions from Stationary Sources," March 18, 1987.
3. Entropy Environmentalists, Inc., "Laboratory Evaluation of a Sampling and
Analysis Method for Hydrogen Chloride Emissions from Stationary Sources:
Interim Report," EPA Contract No. 68-02-4442, Research Triangle Park,
North Carolina, January 22, 1988.
4. U.S. Environmental Protection Agency, "Quality Assurance Handbook for Air
Pollution Measurement Systems, Volume III, Stationary Source Specific
Methods," Publication No. EPA-600/4-77-027b, August 1977.
5. U.S. Environmental Protection Agency, 40 CFR Part 60, Appendix A, Method
6.
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i
i
WIW
HCtOj nm0hif Ma
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METHOD 0051
MIDGET IMPINGER HC1/C1: EMISSION SAMPLING TRAIN
7.1.1.1 Leak-check dry gas metering
system, calculate calibration factor Y.
7.1.1.2 Poat teat calibration
after each teat aeries.
7.1.2 - 7.1.4 Calibrate termometers,
rate meter and barometer.
7.2.1 Set up sample collection train,
connect impmgers in series, and add
reagents. If moisture la to be
determined, weigh Mae Weat impinger
with silica gel.
7.2.2 Perform leak-check on assembled
sampling train.
7.2 3 Immediately prior to sampling,
purge probe for 5 mm. at 2 liter/mm.
7.2.4 Perform aample collection,
adjust aampling rate to 2 litera/min.
for 1 hour. Take readings for dry ges
meter, tempsratura, rate meter, and
vaccum once every 5 mm.
7.3 Stop pump, disconnect tmpmger
sampling tram, quantitatively transfer
contents of impmgers and water rinses
to labeled containers. Retain 50 ml
ah quota of 0.1N H 2SO 4and
NaOH for blanks.
7.4 Calculate ssmpls volume (7.4.2),
volume of wear vapor (7.4.3), total ug
HCL per sample (7.4.4), ug CL 2 per
sample {7.4.5}, and total HCL
concentration (7.4.6).
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METHOD 0100
SAMPLING FOR FORMALDEHYDE AND OTHER CARBONYL COMPOUNDS
IN INDOOR AIR
1.0 SCOPE AND APPLICATION
1.1 This method provides procedures for the sampling of various carbonyl
compounds in indoor air by derivatization with 2,4-dinitrophenylhydrazine (DNPH)
in a silica gel cartridge. The method may be used in conjunction with Method
8315. The following compounds may be sampled by this method:
Compound Name CAS No."
Acetaldehyde 75-07-0
Acetone 67-64-1
Acrolein 107-02-8
Benzaldehyde 100-52-7
Butyraldehyde 123-72-8
Crotonaldehyde 123-73-9
2,5-Dimethylbenzaldehyde 5779-94-2
Formaldehyde 50-00-0
Hexanal 66-25-1
Isovaleraldehyde 590-86-3
Propionaldehyde 123-38-6
m-Tolualdehyde 620-23-5
o-Tolualdehyde 529-20-4
p-Tolualdehyde 104-87-0
Valeraldehyde 110-62-3
8 Chemical Abstract Services Registry Number
1.2 This method is restricted to use by, or under the close supervision
of, trained analytical personnel experienced in sampling organic compounds in
air. Each analyst must demonstrate the ability to generate acceptable results
with this method.
2.0 SUMMARY OF METHOD
2.1 A known volume of indoor air is drawn through a prepacked silica gel
cartridge coated with acidified 2,4-dinitrophenylhydrazine (DNPH), at a
predetermined sampling rate for an appropriate period of time. After sampling,
the sample cartridges are capped and placed in borosilicate glass tubes with
polypropylene caps and placed in cold storage until analysis. The compounds of
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interest may then be eluted from the cartridge with acetonitrile from a plastic
syringe reservoir into a graduated test tube or a volumetric flask.
The eluate is then topped to known volume and refrigerated until analysis.
Analysis may be done High Performance Liquid Chromatography (HPLC), Method 8315,
with an ultraviolet (UV/Vis) detector at 360 nm.
3.0 INTERFERENCES
3.1 Solvents, reagents, glassware and other sample processing may yield
discrete artifacts and/or elevated baselines causing misinterpretation of the
chromatograms. All of these materials must be demonstrated to be free from
interferences, under the conditions of analysis, by analyzing method blanks.
3.1.1 Glassware and plasticware must be scrupulously cleaned.
Clean all glassware and plasticware 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 rinsing with tap water, organic-free
reagent water, and aldehyde-free acetonitrile. After cleaning, glassware
and plasticware should be stored in a clean environment to prevent any
accumulation of dust or other contaminants.
3.1.2 High purity reagents and solvents should be used to minimize
interference problems. Purification of solvents by distillation in all-
glass systems may be necessary.
3.1.3 Polyethylene gloves should be worn when handling the silica
gel cartridges to reduce the possibility of contamination.
3.2 Contamination of the DNPH reagent is a frequently encountered
problem. Formaldehyde, acetone, and 2,4-dinitroanaline (a decomposition product
of DNPH) may be significant analytical impurities in the DNPH reagent at high
concentrations. The DNPH must be purified by multiple recrystallizations in UV-
grade acetonitrile. Recrystallization is accomplished, at 40-60°C, by slow
evaporation of the solvent to maximize crystal size. The purified DNPH crystals
are stored under UV-grade acetonitrile until use. Impurity levels of carbonyl
compounds in the DNPH are determined prior to the analysis of the samples and
should be less than 0.025 /jg/mL. Refer to Sec. 5.9 for a recrystallization
procedure.
3.3 Ozone Interferences - Ozone at high concentration has been shown to
interfere negatively by reacting with both DNPH ar.d its hydrazone derivatives in
the cartridge (Ref. 6).
3.3.1 The extent of interference depends on the temporal variations
of both the ozone and the carbonyl compounds during sampling. The
presence of ozone in the sample stream is readily inferred from the
appearance of new compounds with retention times shorter than that of the
hydrazone of formaldehyde. Figure 1 shows chromatograms of samples of a
formaldehyde-spiked air stream with and without ozone.
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3.3.2 The most direct solution to the ozone interference is to
remove the ozone before the sample stream reaches the cartridge. This
process entails constructing an ozone denuder or scrubber and placing it
on the front of the cartridge. The denuder is constructed out of 1 m of
0.64 cm OD copper tubing, which is filled with a saturated solution of KI
water, allowed to stand for approximately 5 minutes, and dried with a
stream of clean air or nitrogen for about 1 hour. The capacity of the
ozone denuder as described is about 10,000 ppb/hour of ozone. Test
aldehydes that were dynamically spiked into an ambient sample air stream
passed through the denuder with virtually no losses.
3.4 Samples may be contaminated during shipment or storage by diffusion
of volatile organics through the sample bottle septum seal. Field reagent blanks
must be analyzed to determine when sampling and storage procedures have caused
the contamination.
3.5 Matrix interferences may be caused by contaminants acquired by the
sampling process. The extent of matrix interferences will vary considerably from
source to source, depending upon the nature and diversity of the matrix being
sampled. If significant interferences occur due to organic compounds that have
the same retention time, altering the separation conditions by using alternative
HPLC columns or mobile phase conditions may resolve the problem.
4.0 APPARATUS AND MATERIALS
4.1 Sampling Equipment
4.1.1 Sampling System - capable of accurately and precisely
sampling 0.10 to 1.50 L/min of indoor air. The procedures given here
assume use of a dry meter-equipped sampling system operated at flow rates
of at least 0.5 L/min.
NOTE: A normal pressure drop through the sample cartridge approaches 19 kPa at
a sampling rate of 1.5 L/min.
4.1.2 Thermometer and Barometer - to record indoor conditions at
the time of sampling.
4.1.3 Stopwatch - to time sampling.
4.1.4 Rotameters - to allow observation of the flow rate without
interruption of the sampling process.
4.1.5 Mass Flowmeters and Mass Flow Controllers - for metering and
setting the air flow rate through the sample cartridge (0.50 to 1.20
L/min). These are necessary because cartridges have a high pressure drop
and, at maximum flow rates, the cartridge behaves like a "critical
orifice" and can display a flow rate drop over an extended sampling period
(generally less than 5% over a 24 hour period).
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4.1.6 Fittings and Plugs (Luer-Lok or equivalent) - to connect
cartridges to the sampling system and to cap prepared cartridges.
4.1.7 Heated Probe - necessary when the temperature of sampled air
is below 60"C, to insure effective collection of formaldehyde as a
hydrazone.
4.1.8 Silica Gel Cartridges - chromatographic grade, 2 cm x 1.5 cm
ID, with Luer-Lok type fittings on each end, for manual application of
acidified DNPH coating (Sep-PAK from Waters Associates or equivalent).
Commercially pre-packaged pre-coated cartridges are also available
(Thermosorb/F cartridges from Thermedics Inc. or equivalent).
4.2 Glassware
4.2.1 Volumetric Flasks - various sizes, 5 to 2000 mL.
4.2.2 Pipets - various sizes, 1 to 50 mL
4.2.3 Sample Vials
4.2.4 Borosilicate glass culture tubes (20 x 125 mm) with
polypropylene screw caps - for transporting coated cartridges.
4.3 Liquid Syringes (polypropylene are adequate) - 10 mL, used to prepare
DNPH-coated cartridges.
4.4 Syringe Rack - made of an aluminum plate with adjustable legs on all
four corners. Circular holes of a diameter slightly larger than the diameter of
the 10 mL syringes are drilled through the plate to allow batch processing of
cartridges for cleaning, coating, and sample elution. A 0.16 x 36 x 53 cm plate
with 45 holes in a 5x9 matrix is recommended. See Figure 2.
4.5 Cartridge Drying Manifold - has multiple standard male fittings
(Luer-Lok or equivalent). See Figure 2.
4.6 Repetitive Dispensing Pipets - positive displacement, 0 to 10 mL
range, with 1 L reagent bottles (Lab-Industries or equivalent).
4.7 Polyethylene Gloves - used to handle silica gel cartridges.
4.8 Sample Vial Holder - Friction-top metal can (e.g., 4 L paint can) or
a styrofoam box lined with either polyethylene air bubble padding or granular
charcoal to cushion the samples.
4.9 Soap Bubble Flowmeter or Calibrated Wet Test Meter - for calibrating
the sampling flow rate.
4.10 Melting Point Apparatus (optional)
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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 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 Nitrogen gas, N2 - high purity grade.
5.4 Acetonitrile, CH3CN - UV grade.
5.5 Formaldehyde, CH20 - ACS certified or assayed 36.5% solution (W/W).
5.6 Aldehydes and Ketones - analytical grade, used for preparation of
DNPH derivative standards of target analytes other than formaldehyde. See list
on page 1 for possible target analytes.
5.7 Perchloric Acid, HC104 - analytical grade.
5.8 Hydrochloric Acid, HC1 - analytical grade.
5.9 2,4-Dinitrophenylhydrazine (DNPH), C6H6N404 - recrystallize at least
twice with UV grade acetonitrile using the following procedure:
NOTE: This procedure should be performed under a properly ventilated hood.
Inhalation of acetonitrile can result in nose and throat irritation (brief
exposure at 500 ppm) or more serious effects at higher concentration
and/or longer exposures.
5.9.1 Prepare a saturated solution of DNPH by boiling excess DNPH
in 200 ml of acetonitrile for approximately 1 hour.
5.9.2 After 1 hour, remove and transfer the supernatant to a
covered beaker on a hot plate and allow gradual cooling to 40 to 60°C.
Maintain this temperature range until 95% of the solvent has evaporated
leaving crystals.
5.9.3 Decant the solution to waste and rinse the remaining crystals
twice with three times their apparent volume of acetonitrile.
5.9.4 Transfer the crystals to a clean beaker, add 200 ml of
acetonitrile, heat to boiling, and again let the crystals grow slowly at
40 to 60°C until 95% of the solvent has evaporated. Repeat the rinsing
process as in Sec. 5.9.3.
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5.9.5 Take an aliquot of the second rinse, dilute 10 times with
acetonitrile, acidify with 1 ml of 3.8 M perchloric acid per 100 ml of
DNPH solution, and analyze by HPLC Method 8315). An acceptable impurity
level is less than 0.025 mg/L of formaldehyde in recrystallized DNPH
reagent or below the sensitivity (ppb, v/v) level indicated in Table 1 for
the anticipated sample volume.
5.9.6 If the impurity concentration is not satisfactory, pipet off
the solution to waste, repeat the recrystallization as in Sec. 5.9.4 but
rinse with two 25 ml portions of acetonitrile. Prep and analyze the
second rinse as in Sec. 5.9.5.
5.9.7 When the impurity concentration j_s satisfactory, place the
crystals in an all-glass reagent bottle, add another 25 mL of
acetonitrile, stopper, and shake the bottle. Use clean pipets when
removing the saturated DNPH stock solution to reduce the possibility of
contamination of the solution. Maintain only a minimum volume of the
saturated solution adequate for day to day operation to minimize waste of
the purified reagent.
5.10 Refer to the determinative method (Method 8315) for procedures
regarding the preparation of DNPH derivatives, standards of the derivatives, and
calibration standards for HPLC analysis. All standard solutions should be stored
at about 4°C in a glass vial with a Teflon®-!ined cap, with minimum headspace,
and in the dark. They should be stable for about 6 weeks. All standards should
be checked frequently for signs of degradation or evaporation, especially just
prior to preparing calibration standards from them.
5.11 Preparation of DNPH-Coated Sep-PAK Cartridges (if pre-packaged pre-
coated cartridges, as in Sec. 4.1.8, are not used)
NOTE: This procedure must be performed in an atmosphere with a very low aldehyde
background. The atmosphere above the acidified solution should preferably
be filtered through a DNPH-coated silica gel cartridge to minimize
contamination from laboratory air. All glassware and plasticware must be
scrupulously cleaned and rinsed with deionized water and aldehyde free
acetonitrile. Contact of reagents with laboratory air must be minimized.
Polyethylene gloves must be worn when handling the cartridges.
5.11.1 DNPH Coating Solution
5.11.1.1 Pipet 30 ml of saturated DNPH stock solution into a
1000 mL volumetric flask, add 500 mL acetonitrile, and acidify with
1.0 mL of concentrated HC1.
5.11.1.2 Shake solution and dilute to volume with
acetonitrile. Stopper the flask, invert, and shake several times
until the solution is homogeneous. Transfer the acidified solution
to a reagent bottle equipped with a 0 to 10 mL range repetitive
pipet dispenser. Prime the dispenser and slowly dispense 10 to 20
mL to waste.
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5.11.1.3 Dispense an aliquot solution to a sample vial, and
check the impurity level of the acidified solution by HPLC according
to Sec. 7.2.
5.11.1.4 The impurity concentration should be less than 0.025
jug/mL formaldehyde, similar to that in the DNPH stock solution.
5.11.2 Coating of Sep-PAK Cartridges
5.11.2.1 Open the Sep-PAK package, connect the short end to
a 10 ml syringe and place it in the syringe rack. The syringe rack
used for coating and drying the sample cartridges is illustrated in
Figures 2(a) and 2(b).
5.11.2.2 Using a positive displacement, repetitive pipet, add
10 ml of acetonitrile to each of the syringes.
5.11.2.3 Let the liquid drain to waste by gravity. Remove
any air bubbles that may be trapped between the syringe and the
silica cartridge by displacing them with the acetonitrile in the
syringe.
5.11.2.4 Once the effluent flow at the outlet of the
cartridge has stopped, dispense 7 ml of the acidified DNPH coating
reagent into each of the syringes using the repetitive pipet
dispenser.
5.11.2.5 Let the coating reagent drain by gravity through the
cartridge until flow at the other end of the cartridge stops.
5.11.2.6 Wipe the excess liquid at the outlet of each of the
cartridges with clean tissue paper.
5.11.2.7 Assemble a drying manifold as shown in Figure 2(b).
This contains a previously prepared, DNPH-coated, cartridge at each
of the exit ports (e.g., these scrubber or "guard cartridges" can be
prepared by drying a few of the newly coated cartridges as per the
following sections, and "sacrificing" these few to assure the purity
of the rest). The "guard cartridges" serve to remove traces of
formaldehyde that may be present in the nitrogen gas supply.
5.11.2.8 Insert cartridge connectors (flared at both ends,
0.64 cm OD x 2.5 cm Teflon® FEP tubing with ID slightly smaller than
the OD of the cartridge port) onto the long end of the scrubber
cartridges.
5.11.2.9 Remove the cartridges from the syringes and connect
the short ends of the cartridges to the open end of the cartridge
connectors already attached to the scrubber cartridges.
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5.11.2.10 Pass nitrogen through each of the cartridges at
about 300 to 400 mL/min.
5.11.2.11 Rinse the exterior surfaces and outlet end of the
cartridges with acetonitrile using a Pasteur pipet.
5.11.2.12 After 15 minutes, stop the flow of nitrogen, wipe
the cartridge exterior free of rinse acetonitrile and remove the
dried cartridges.
5.11.2.13 Plug both ends of the coated cartridge with
standard polypropylene Luer-Lok male plugs and place the plugged
cartridge in a borosilicate glass culture tube with polypropylene
screw caps.
5.11.2.14 Put a serial number and a lot number label on each
of the individual cartridge glass storage containers and refrigerate
the prepared lot until use. Cartridges will maintain their
integrity for up to 90 days stored in refrigerated, capped culture
tubes.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Assemble the sampling system, and ensure that the pump is capable of
constant flow rate throughout the sampling period. The coated cartridges can be
used as direct probes and traps for sampling indoor air when the temperature is
above 60°F.
6.1.1 If the temperature in below 60°F, use the heated probe,
mentioned in Sec. 4.1.7, to warm the air entering the sampling equipment.
6.1.2 If necessary, add an ozone denuder (see Sec. 3.3).
6.2 Before sample collection, check the system for leaks. Plug the input
(short end) of the cartridge so no flow is indicated at the output end of the
pump. The mass flowmeter should not indicate any air flow through the sampling
apparatus.
6.3 Install the entire assembly (including a "dummy" sampling cartridge)
and check the flow rate at a value near the desired rate. In general, flow rates
of 500-1200 mL/min should be employed. The total moles of carbonyl in the volume
of air sampled should not exceed that of the DNPH (2 mg or 0.01 mmole/cartridge).
In general, a safe estimate of the sample size should be approximately 75% of the
DNPH loading of the cartridge (approximately 200 /zg as HCHO). Generally,
calibration is accomplished using a soap bubble flowmeter or calibrated wet test
meter connected to the flow exit, assuming the system is sealed.
NOTE: ASTM Method D3686 describes an appropriate calibration scheme that does
not require a sealed flow system downstream of the pump.
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6.4 Ideally, a dry gas meter is included in the system to measure and
record total flow. If a dry gas meter is not available, the operator must
measure and record the sampling flow rate at the beginning and end of the
sampling period to determine sample volume. If the sampling period exceeds two
hours, the flow rate should be measured at intermediate points during the
sampling period. Include a rotameter to allow observation of the flow rate
without interruption of the sampling process.
6.5 Before sampling, remove the glass culture tube from the friction-top
metal can or styrofoam box. Let the cartridge warm to room temperature in the
glass tube before connecting it to the sample train.
6.6 Using polyethylene gloves, remove the coated cartridge from the glass
tube and connect it to the sampling system with a Luer adapter fitting. Seal the
glass tube for later use, and connect the cartridge to the sampling train so that
the short end becomes the sample inlet.
6.7 Turn the sampler on, record the start time, and adjust the flow to
the desired rate. A typical flow rate through one cartridge is 1.0 L/min and 0.8
L/min for two cartridges in tandem.
6.8 Operate the sampler for the desired period, with periodic recording
of the sampling variables such as sample flow rate, pressure, and temperature.
6.9 At the end of the sampling period, stop the flow and record the stop
time. If a dry gas meter or equivalent is not used, the flow rate must be
checked just before stopping the flow. The average sample flow rate may be
calculated using the equation in Sec. 9.1.1. If the flow rate at the beginning
and end of the sampling period differ by more than 15%, the sample should be
marked as suspect.
6.10 Immediately after sampling, remove the cartridge (using polyethylene
gloves) from the sampling system, cap with Luer end plugs, and place it back in
the original labeled glass culture tube. Cap the culture tube, seal it with
Teflon® tape, label the tube, and place it in a friction-top can containing 2-5
cm of granular charcoal or styrofoam box with appropriate padding. Refrigerate
the culture tubes until analysis. The refrigeration period prior to analysis
should not exceed 30 days.
NOTE: If samples are to be shipped to a central laboratory for analysis, the
duration of the non-refrigerated period should be kept to a minimum,
preferably less than two days.
6.11 Use the equations found in Sees. 9.1.2 and 9.1.3 to calculate the
total volume of air sampled and the total volume of air sampled at standard
conditions.
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7.0 SAMPLE RECOVERY
7.1 The samples are returned to the laboratory in a friction-top can
containing 2 to 5 cm of granular charcoal and stored in a refrigerator until
analysis. Alternatively, the samples may also be stored alone in their
individual glass containers. The time between sampling and analysis should not
exceed 30 days.
7.2 Refer to the determinative method (Method 8315) for procedures
regarding desorption of the sample from the cartridge and HPLC analysis
preparation.
8.0 CALIBRATIONS
8.1 Refer to Sec. 6.0 for requirements regarding the calibration of the
sampling system flow rate and equipment for the determination of total flow.
8.2 Refer to the determinative method for procedures regarding
calibration of the HPLC analysis system.
8.3 Barometer - Adjust the barometer initially and before each test
series to agree within ±2.5 mm Hg (±0.1 in Hg) of the mercury barometer or the
corrected barometric pressure value reported by a nearby National Weather Service
Station (same altitude above sea level). Note that adjustment for elevation
differences between the weather station and the sampling point is applied at a
rate of minus 2.5 mm Hg (0.1 in Hg) per 30 m (100 ft) elevation increase.
8.4 Thermometers
8.4.1 If a mercury-in-glass reference thermometer is to be used, it
must conform to ATSM E-l 63C or 63F specifications.
8.4.2 If a thermocouple is to be used, it must be calibrated in the
laboratory according to the manufacturer's specifications. The
calibration should be done both with and without the use of any extension
leads.
9.0 CALCULATIONS
9.1 Calculation of the total volume of air sampled at standard conditions.
9.1.1 If a dry gas meter or equivalent total flow indicator is not
used, the average sample flow rate, FRave in mL/minute, may be calculated
using the following equation:
rr\i •+• ri\2 + ... + rKpj
r K.,..- =
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where:
FR,, FR2, ..., FRN = Flow rates determined at the beginning,
end, and intermediate points during
sampling
N = Number of flow rates averaged
9.1.2 The total volume of air sampled at the measured temperature
and pressure, VTot in liters (L), may be calculated using the following
equation:
(Time2 - Time,) (FRave)
1,000
where:
Time2 = Stop time (min)
Time! = Start time (min)
(Time2 - Time.,) = Total sampling time (min)
FRave = Average flow rate (mL/min)
9.1.3 The total volume of air sampled converted to standard
conditions, VTotStd in liter (L) at 25°C and 101.3 kPa, may be calculated
using the following equation:
Pave 298°C
V = V " "
* TotStd ' Tot
101.3 kPa (273°C + Tave)
where:
VTot = Total sample volume (L) at measured temperature and
pressure
Pa*e = Average indoor pressure (kPa)
Lve = Average indoor temperature (°C)
10.0 DETERMINATION OF VOLUME TO BE SAMPLED
10.1 Refer to Table 1 for information regarding method "sensitivity" at
various sampling volumes.
11.0 QUALITY CONTROL
11.1 Refer to Chapter One for quality control procedures.
11.2 Method Blanks - A method blank must be prepared for each set of
analytical operations, to evaluate contamination and artifacts that can be
derived from glassware, reagents, and sample handling in the laboratory.
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11.3 Field Blanks - Field blanks must be submitted with the samples at
each sampling site or 10% of the field samples, whichever is larger, should be
shipped and analyzed with each group of samples. The field blank is treated
identically to the samples except that no air is drawn through the cartridge.
It is desirable to analyze blank cartridges retained in the laboratory (method
blanks) as well, to distinguish between possible field and laboratory
contamination.
11.4 Blank and Matrix Spikes - A procedure for spiking air sampling
cartridges is not yet established for this sampling technique. Refer to
Attachment A for information regarding possible techniques for accomplishing
sample spiking. Proper QC procedures require that a blank spike and matrix spike
be processed for each batch of 10 samples or less. As the MDL becomes better
established for this method, the representative spike concentration should be set
at 10 times the MDL, for that matrix, to account for interferences.
12.0 METHOD PERFORMANCE
12.1 The method detection limit (MDL) is defined as the minimum
concentration of the test compound that can be measured and reported with 99
percent confidence as being greater than zero. The MDL actually achieved in a
given analysis will vary, as it is dependent on instrument sensitivity and matrix
effects. The MDLs for the target analytes in the method have not yet been
established.
12.2 Table 1 illustrates the sensitivity for the target analytes of
interest found in ambient air that have been identified using two Zorbax CDS
columns in series.
13.0 REFERENCES
1. Winberry, Jr., W.T., Murphy, N.T., and Riggin, R.M., Method TO-11,
Compendium of Methods For the Determination of Toxic Organic Compounds in
Ambient Air, EPA-600/6-89-017, U.S. Environmental Protection Agency,
Research Triangle Park, NC, June 1988.
2. Tejada, S.B., "Standard Operating Procedure for DNPH-coated Silica
Cartridges For Sampling Carbonyl Compounds in Air and Analysis by High-
performance Liquid Chromatography," Unpublished, U.S. Environmental
Protection Agency, Research Triangle Park, NC, March 1986.
3. Tejada, S.B., "Evaluation of Silica Gel Cartridges Coated in situ with
Acidified 2,4-Dinitrophenylhydrazine for Sampling Aldehydes and Ketones in
Air," Intern. J. Environ. Anal. Chem., Vol. 26:167-185, 1986.
4. Quality Assurance Handbook for Air Pollution Measurement Systems, Volume
II - Ambient Air Specific Methods, EPA-600/4-77-027A, U.S. Environmental
Protection Agency, Research Triangle Park, NC, July 1979.
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5. Riggin, R.M., Technical Assistance Document for Sampling and Analysis of
Toxic Organic Compounds in Ambient Air, EPA-600/4-83-027, U.S.
Environmental Protection Agency, Research Triangle Park, NC, June, 1983.
6. Arnts, R.R. and Tejada, S.B., "2,4-Dinitrophenylhydrazine-Coated Silica
Gel Cartridge Method for Determination of Formaldehyde in Air", Env. Sci.
and Tech. 23, 1428-1430 (1989).
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TABLE 1
SENSITIVITY (ppb, v/v) OF SAMPLING AND ANALYSIS FOR ALDEHYDES AND
KETONES IN AMBIENT AIR USING AN ADSORBENT CARTRIDGE
FOLLOWED BY GRADIENT HPLC"
Compound
Acetaldehyde
Acetone
Acrolein
Benzaldehyde
Butyraldehyde
Crotonaldehyde
2,5-Dimethyl-
benzaldehyde
Formaldehyde
Hexanal
Isovaleraldehyde
Propionaldehyde
m-Tolualdehyde
o-Tolualdehyde
p-Tolualdehyde
Valeraldehyde
10
Sample Volume (L)k
20 30
40
50 100 200 300 400 500
1.36
1.28
1.29
1.07
1.21
1.22
0.68
0.64
0.65
0.53
0.61
0.61
0.45
0.43
0.43
0.36
0.40
0.41
0.34
0.32
0.32
0.27
0.30
0.31
0.27
0.26
0.26
0.21
0.24
0.24
0.
0.
0.
0.
0.
0.
14
13
13
11
12
12
0.07
0.06
0.06
0.05
0.06
0.06
0.05
0.04
0.04
0.04
0.04
0.04
0
0
0
0
0
0
.03
.03
.03
.03
.03
.03
0.03
0.03
0.03
0.02
0.02
0.02
0.97
1.45
1.09
1.15
1.28
1.02
1.02
1.02
1.15
0.49
0.73
0.55
0.57
0.64
0.51
0.51
0.51
0.57
0.32
0.48
0.36
0.38
0.43
0.34
0.34
0.34
0.38
0.24
0.36
0.27
0.29
0.32
0.25
0.25
0.25
0.29
0
0
0
0
0
0
0
0
0
.19
.29
.22
.23
.26
.20
.20
.20
.23
0.10
0.15
0.11
0.11
0.13
0.10
0.10
0.10
0.11
0.05
0.07
0.05
0.06
0.06
0.05
0.05
0.05
0.06
0.
0.
0.
0.
0.
0.
0.
0.
0.
03
05
04
04
04
03
03
03
04
0.02
0.04
0.03
0.03
0.03
0.03
0.03
0.03
0.03
0.02
0.03
0.02
0.02
0.03
0.02
0.02
0.02
0.02
The ppb values are measured at 1 atm and 25°C. The sample cartridge is
eluted with 5 mL acetonitrile and 25 p.1 is injected into the HPLC. The
maximum sampling flow through a DNPH-coated Sep-PAK is about 1.5 L/minute.
b A sample volume of 1000 L was also performed. The
sensitivity of 0.01 ppb for all the target analytes.
results show a
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FIGURE 1
CARTRIDGE SAMPLES OF A FORMALDEHYDE AIR STREAM
WITH (A) AND WITHOUT (B) OZONE
8
x 0
I
4 6
Tlmt, mln
B
10
x * unknown
0-DNPH
1 » formaldehyde
2 * acetaldehyde
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FIGURE 2
SYRINGE RACKS FOR COATING AND DRYING SAMPLE CARTRIDGES
10 ml Glass
Syringe
Adsorbent
Tube
(a) RACK FOR COATIN" CARTRIDGES
Test Tube
Rack
Waste
Beaker
Luer-Lok
Fitting "~
«_ «_N Gas Stream
Adsorbent
Tubes
Waste
ViaJ
^^B
^1^
u
(b) RACK FOR DRYING DNPH-COATED CARTRIDGES
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ATTACHMENT A
This method does not contain a procedure for spiking cartridges for blank
spikes and matrix spikes to determine percent recovery. Two suggested techniques
for spiking cartridges are as follows:
1) A spike may be performed by introducing an aliquot of a solution
containing the target analytes by pipet or syringe directly onto a
cartridge in the field or in the laboratory. Standard spike and recovery
procedures are followed and the field spike sample is returned to the
laboratory for analysis. An aliquot of the field spike standard is
retained in the laboratory for derivatization and comparative analysis.
2) Another technique would include spiking the sampling cartridge using a TGM
555 analyzer which produces gaseous formaldehyde standards. However, it
should be noted that the procedures required to produce accurate, dynamic,
low-level standard mixtures of organics in air are non-routine. The
techniques developed for use in evaluating other air sampling procedures
employ a 3-stage dynamic gas dilution system coupled with a constant-rate
vapor generation assembly containing a trioxane permeation tube (VICI
Medtronics Dynacal permeation device or equivalent) that is maintained at
55°C. Trioxane vapor is converted stoichiometrically to formaldehyde
vapor using a special high-temperature (160"C) catalytic converter
assembly. This method of sample introduction has been used when testing
continuous sampling apparatus.
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* U.S. GOVERNMENT PRINTING OFFICE:! 995-386-824/33253 JanU3ry 1995
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