5551
905R85112
Method 680. Determination of Pesticides and PCBs
in Water and Soil/Sediment
by Gas Chromatography/Mass Spectrometry
November 1985
Ann Alford-Stevens
Thomas A. Bellar
James W. Eichelberger
William L. Budde
Physical and Chemical Methods Branch
Environmental Monitoring and Support Laboratory
Office of Research and Development
U. S. Environmental Protection Agency
Cincinnati, Ohio 45268
US. Environmental Protection Agency
Region V, Library
230 South Dearborn Street
Chicago, HHnois 60604
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U,S. Environmental Protection Agency^
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INDEX
Section
Number Subject
1 Scope and Application
2 Summary of Method
3 Definitions
4 Interferences
5 Safety
6 Apparatus and Equipment
7 Reagents and Consumable Materials
8 Sample Collection, Preservation and Handling
9 Calibration
10 Quality Control
11 Procedures
12 Calculations
13 Automated Identification and Measurement
14 Method Perforamnce •
15 References
Tables
1
2
3
4
5a
5b
6
7a
7b
7c
8
9
10
11
12
13
14
15
Recommended GC Operating Conditions
PCS Congeners Used as Calibration Standards
Scheme for Preparation of PCB Stock Solution
Composition and Approximate Concentrations of Calibration Solutions
for Full-Range Data Acquisition
Composition and Approximate Concentrations of Calibration Solutions
for SIM Data Acquisition for PCB Determinations
Composition and Approximate Concentrations of Calibration Solutions
for SIM Data Acquisition for Pesticide Determinations
Criteria for DFTPP Spectrum
Ions for Selected Ion Monitoring to Determine PCBs by Acquiring
Data for Four Sets of £35 Ions Each
Ions for Selected Ion Monitoring to Determine PCBs by Acquiring
Data for Five Sets of £20 Ions Each
Five Ion Sets of £20 Ions Each for Selected Ion Monitoring of PCBs
Retention Time Data For PCB Isomer Groups and Calibration Congeners
Ions for Selected Ion Monitoring Data Acquisition for Pesticide
Analytes, Internal Standards and Surrogate Compounds
Ion Sets for Selected Ion Monitoring of Pesticide Analytes,
Internal Standards, and Surrogate Compounds
Known Relative Abundances of Ions in PCB Molecular Ion Clusters
Quantitation, Confirmation, and Interference Check Ions for
PCB Analytes, Internal Standards, and Surrogate Compounds
Correction for Interference of PCB Containing Two Additional Chlorines
Correction for Interference of PCB Containing One Additional Chlorine
Accuracy and Precision of Automated Measurements of PCBs and Pesticides
in Fortified Water Extracts
Figures
1
2
Total ion current profile of PCB calibration congeners and
pesticide Analytes
Diagram indicating approximate relative retention times of PCB
isomer groups and retention time congeners.
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1. SCOPE AND APPLICATION
1.1. This method provides procedures for mass spectrometric determination
of polychlorinated biphenyls (PCBs) and the listed pesticides in water,
soil, or sediment. This method is applicable to samples containing PCBs
as single congeners or as complex mixtures, such as commercial Arodors.
PCBs are identified and measured as isomer groups (i.e., by level of
chlorination). The existence of 209 possible PCS congeners makes
impractical the listing of the Chemical Abstracts Service Registry
Number (CASRN) for each potential method analyte. Because PCBs are
identified and measured as isomer groups, the non-specific CASRN for
each level of chlorination is used to describe method analytes.
Analyte(s)
Aldrin
BHCs
alpha isomer
beta isomer
delta isomer
gamma isomer (lindane)
Chlordane ( technical )
alpha-chlordane
gamma-chlordane
trans-nonachlor
4,4'-DDD
4, 4 '-DDE
4, 4 '-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Heptachlor
Heptachlor epoxide
Methoxychlor
PCBs
Monochlorobiphenyls
Dichlorobiphenyls
Trichlorobiphenyls
Tetrachlorobiphenyls
Pentachlorobiphenyls
Hexachlorobiphenyls
Heptachlorobiphenyls
Octachlorobiphenyls
Nonachlorobiphenyls
Decachlorobiphenyl
Formula
C12H8C16
CgHgClg
CgHgClg
CgHgClg
C14H10C14
C14H8C14
C14H9C15
C12H8C160
CgHgClgO3S
C8HgClgO3S
CgH4Clg04S
C12H8ClgO
C-]2H8ClgO
C-|2H8C12
C-|2H7Cl3
C12HgCl4
C^2H5Cl5
Ci2H4Clg
CASRN
309-00-2
319-84-6
319-85-7
319-86-8
58-89-9
57-74-9
5103-71-9
5103-74-2
19765-80-5.
72-54-8
72-55-9
50-29-3
60-57-1
959-98-8
33213-65-9
1031-07-8
72-20-8
7421-93-4
53494-70-5
76-44-8
1024-57-3
72-43-5
C-|2H2C18
Ci2HClg
27323-
25512-
25323-
26914-
25429-
26601-
28655-
31472-
53742-
2051-
•18-8
•42-9
68-6
33-0
•29-2
•64-9
•71-2
83-0
•07-7
•24-3
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-2-
1.2 Detection Limits vary among method analytes and with sample matrix, sample
preparation procedures, condition of the GC/MS system, type of data
acquisition, and individual samples. The calculated method detection
limit (MDL) for each pesticide in fortified reagent water extracts analyzed
with full-range data acquisition is presented in Sect. 14. Analysis of
calibration solutions indicated that the calculated MDLs do not accurately
reflect instrumental detection limits. The following guidance is based on
numerous analyses of calibration solutions with one instrument over a period
of approximately six months. Pesticide analytes other than endosulfans
I and II can be identified and accurately measured when the injected
aliquot contains 2 ng of each analyte; the endosulfans require about 4 ng
each. With selected-ion-monitoring (SIM) data acquisition, pesticide
analyte detection limits are lowered by at least a factor of five. Detection
limits for individual PCS congeners increase with increasing number of
chlorine atoms, with the detection limit for decachlorobiphenyl being
about 5-10 times higher than that of a monochlorobiphenyl. A monochloro-
biphenyl can be identified and accurately measured when the injected
extract aliquot contains 1 ng and full-range data are acquired. The
detection limit for total PCBs will depend on the number of individual
PCS congeners present. SIM data acquisition procedures reduce the detection
limit for PCBs by at least a factor of three.
2. SUMMARY OP METHOD
A 1-L water sample is placed in a separatory funnel and extracted with methylene
chloride. Appropriate extraction procedures for soil/sediment samples will be
added when results are obtained from ongoing experiments. The extract is dried
and exchanged to hexane during concentration to a final volume of 1 mL or less.
Sample extract components are separated with capillary column gas chromatography
(GC) and identified and measured with low resolution, electron ionization mass
spectrometry (MS). An interfaced data system (DS) to control data acquisition
and to store, retrieve, and manipulate mass spectral data is essential. Either
full-range or selected-ion-monitoring (SIM) data are acquired, depending on the
concentration range of concern. If full-range data are acquired, all method
analytes can be identified and measured with one GC/MS analysis. If all pesti-
cides and PCBs must be determined and if SIM data are necessary to meet required
detection limits, two GC/MS analyses are necessary, one to detect and measure
pesticides and one to detect and measure PCBs.
Two surrogate compounds are added to each sample before sample preparation;
these compounds are C12~4»4'~DDT and 3Cg-gamma-BHC. Two internal standards,
chrysene-d-) 2 **& phenanthrene-d^ g / are added to each sample extract before GC/MS
analysis and are used to calibrate MS response. Each concentration measurement
is based on an integrated ion abundance of one characteristic ion. All pesticides
are identified as individual compounds, and a concentration is calculated by
relating the MS response of each compound to the MS response of the internal
standard with GC retention time nearer that of the pesticide analyte. The
extent of sample contamination with technical chlordane is indicated by identi-
fication and measurement of the two most persistent components, gamma-chlordane
and nonachlor. (Alpha-chlordane and heptachlor, other major components of
technical chlordane, may also be present and will be detected and measured
with this method.)
PCBs are identified and measured as isomer groups (i.e., by level of chlorination).
A concentration is measured for each PCB isomer group; total PCB concentration
in each sample extract is obtained by summing isomer group concentrations.
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-3-
Nine selected PCS congeners are used as calibration standards/ and one internal
standard, chrysene-d-^/ is used to calibrate MS response to PCBs, unless sample
conditions require the use of the second internal standard, phenanthrene-dig.
3. DEFINITIONS
3.1 CONCENTRATION CALIBRATION SOLUTION (CAL) — A solution of method analytes
used to calibrate the mass spectrometer response.
3.2 CONGENER NUMBER -- Throughout this method, individual PCBs are described
with the number assigned by Ballschmiter and Zell (2). (This number is
also used to describe PCS congeners in catalogs produced by Ultra Scientific,
Rope, RI.)
3.3 INTERNAL STANDARD -- A pure compound added to a sample extract in known
amounts and used to calibrate concentration measurements of other compounds
that are sample components. The internal standard must be a compound
that is not a sample component.
3.4 LABORATORY DUPLICATES (LD1 and LD2) —Two sample aliquots taken in the
analytical laboratory are analyzed with identical procedures. Analysis
of laboratory duplicates indicates precision associated with laboratory
procedures but not with sample collection, preservation or storage procedures.
3.5 LABORATORY PERFORMANCE CHECK SOLUTION (LPC) — A solution of method analytes,
surrogate compounds, and internal standards used to evaluate the performance
of the GC/MS/DS with respect to a defined set of method criteria.
3.6 LABORATORY REAGENT BLANK (LRB) — An aliquot of reagent water or neutral
solid reference material that is treated as a sample. It is exposed to
all glassware and apparatus, and all method solvents, reagents, internal
standards, and surrogate compounds are used. The extract is concentrated
to the final volume used for samples and is analyzed the same as a sample
extract.
3.7 LABORATORY SPIKE DUPLICATE SAMPLE — One aliquot (LSD) of a sample is
analyzed before fortification with any method analytes. In the laboratory,
a known quantity of method analytes (LSA) is added to two independent
aliquots of the same sample, and final analyte concentrations (LF1 and
LF2) are measured with the same analytical procedures used to measure LSD.
3.8 LABORATORY SURROGATE SPIKE
3.S.1 Measured Value (LS1) — Surrogate compound concentration measured
with the same procedures used to measure sample components.
3.8.2 Theoretical Value (LS2) ~ The concentration of surrogate compound
added to a sample aliquot before extraction. This value is determined
from standard gravimetric and volumetric techniques used during
sample fortification.
3.9 METHOD DETECTION LIMIT (MDL) — A statistically determined value (1)
indicating the minimum concentration of an analyte that can be identified
and measured in a sample matrix with 99% confidence that the analyte
concentration is greater than zero. This value varies with the precision
of the replicate measurements used for the calculation.
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3.10 PERFORMANCE EVALUATION SAMPLE — A sample containing known concentrations
of method analytes that has been analyzed by multiple laboratories to
determine statistically the accuracy and precision that can be expected
when a method is performed by a competent analyst. Analyte concentrations
are unknown to the analyst.
•'
3.11 QUALITY CONTROL (QC) CHECK SAMPLE — A sample containing known concentra-
tions of analytes that is analyzed by a laboratory,to demonstrate that it
can obtain acceptable identifications and measurements with procedures to
be used to analyze environmental samples containing the same or similar
analytes• Analyte concentrations are known by the analyst. Preparation
of the QC check sample by a laboratory other than the laboratory performing
the analysis is highly desirable.
3.12 SURROGATE COMPOUND — A compound not expected to be found in the sample
is added to a sample aliquot before extraction and is measured with the
same procedures used to measure sample components. Associated with the
surrogate compound are two values, laboratory surrogate spike- measured
value (LS1) and laboratory surrogate spike - theoretical value (LS2).
The purpose of a surrogate compound is to monitor method performance
with each sample.
4. INTERFERENCES
4.1 Interferences may be caused by contaminants in solvents/ reagents, glassware,
and other sample processing equipment. Laboratory reagent blanks (LRBs)
are analyzed routinely to demonstrate that these materials are free of
interferences under the analytical conditions used for samples.
4.2 To minimize interferences, glassware (including sample bottles) should
be meticulously cleaned. As soon as possible after use, rinse glassware
with the last solvent used. Then wash with detergent in hot water and
rinse with tap water followed by distilled water. Drain dry and heat in a
muffle furnace at 450°C for a few hours. After cooling, store glassware
inverted or covered with aluminum foil. Before using, rinse each piece
with an appropriate solvent. Volumetric glassware should not be heated
in a muffle furnace.
4.3 For both pesticides and PCBs, interference can be caused by the presence
of much greater quantities of other sample components that overload the
capillary column; additional sample extract preparation procedures must
then be used to eliminate interferences. Capillary column GC retention
times and the compound-specific characteristics of mass spectra eliminate
many interferences that formerly were of concern with pesticide/PCB
determinations with electron capture detection. The approach and identi-
fication criteria used in this method for PCBs eliminate interference by
most chlorinated compounds other than other PCBs. With the isomer group
approach, coeluting PCBs that contain the same number of chlorines are
identified and measured together. Therefore, coeluting PCBs are a problem
only if they contain a different number of chlorine atoms. This interference
problem is obviated by rigorous application of the identification criteria
described in this method.
4.4 For SIM identification and measurement of pesticides, other chlorinated
sample components that produce the same quantitation and confirmation
ions may interfere, but only if retention times are nearly equivalent.
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-5-
5. SAFETY
5.1 The toxlcity or carcinogenicity of each chemical used in this method
has not been precisely defined. Therefore, each should be treated as a
potential health hazard, and exposure should be reduced to the lowest
feasible level. Each laboratory is responsible for safely disposing
materials and for maintaining awareness of OSHA regulations regarding
safe handling of the chemicals used in this method. A reference file of
material data handling sheets should be made available to all personnel
involved in analyses. Additional information on laboratory safety is
available (3-5).
5.2 The following method analytes have been classified as known or suspected
human or mammalian carcinogens: BHCs, 4,4'-ODD, 4,4'-DDT, and PCBs.
Primary standards of these compounds should be prepared in a hood. A
toxic gas respirator should be worn when the analyst handles solutions
containing high concentrations of these compounds.
t
6. APPARATUS AND EQUIPMENT
6.1 SAMPLING EQUIPMENT
6.1.1 Water Sample Bottles — Meticulously cleaned (Sect. 4.2) 1-L or
1-qt amber glass fitted with a. Teflon-lined screw cap. (Bottles in
which high purity solvents were received can be used as sample
bottles without additional cleaning if they have been handled
carefully to avoid contamination during and after use of original
contents.)
6.1.2 Soil/Sediment Sample Bottles — Appropriate containers will be
specified when appropriate extraction procedures are determined.
6.2 GLASSWARE
6.2.1 Separatory Funnel ~ 2-L with Teflon stopcock.
6.2.2 Drying Column — glass column approximately 400 mm long X 19 mm ID
with coarse frit filter disc.
6.2.3 Chromatography Column — glass column approximately 400 mm long
X 19 mm ID with coarse frit filter disc and Teflon stopcock.
6.2.4 Concentrator Tube —- 10-mL graduated Kuderna-Danish design
with ground-glass stopper.
6.2.5 Evaporative Flask — 500-mL Kuderna-Danish design that is
attached to concentrator tube with springs.
6.2.6 Snyder Column — three-ball macro Kuderna-Danish design.
6.2.7 Vials — 10- to 15-mL amber glass with Teflon-lined screw caps.
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-6-
6.3 COMPUTERIZED GC/MS SYSTEM
6*3.1 The GC must be capable of temperature programming and be equipped
with all required accessories, such as syringes, gases, and a capillary
column. The GC injection port must be designed for capillary columns.
Manual splitless injections were used to acquire data used as the basis
for quality control requirements. An automatic injector, however, is
desirable, because it should provide more precise retention times and
areas. On-column injection with an uncoated precolumn is encouraged,
because high mass descrimination and analyte degradation problems
are minimized with this technique. With some GCs, however, the
irreproducibility of the low initial column temperature required for
on-column injections will cause irreproducible retention times (RTs)
and relative retention times (RRTs). That can result in an inability
to distinguish between two closely-eluting pesticide isomers and may
cause ion sets to be changed at inappropriate times during SIM data
acquistion. Splitting injections are not recommended.
6.3.2 Either full range or SIM mass spectral data are obtained with electron
ionization at a nominal electron energy of 70 eV. To ensure sufficient
precision of mass spectral data, the required MS scan rate must
allow acquisition of at least five full-range mass spectra or five
data points for each monitored ion while a sample component elutes
from the GC. The MS must produce a mass spectrum meeting all criteria
for £20 ng of decafluorotriphenylphosphine (DFTPP) introduced through
the GC inlet.
6.3.3 An interfaced data system (OS) is required to acquire, store, reduce,
and output mass spectral data. The OS must be capable of searching
a data file for specific ions and plotting ion abundances versus time
or spectrum number to produce selected ion current profiles (SICPs)
and extracted ion current profiles (EICPs). Also required is the
capability to obtain chromatographic peak areas between specified
times or spectrum numbers in SICPs or EICPs. Total data acquisition
time per cycle should be £0.5 s and must not exceed 1.5 s.
6.3.4 SIM Option — For SIM data acquisition, the DS must be equipped with
software capable of acquiring data for multiple groups of ions,
and the DS must allow automated and rapid changes of the set of ions
being monitored. To acquire all PCS data needed for implementation
of two currently-available automated interpretation procedures, the
SIM program must be capable of acquiring data for four groups (or
mass ranges) each consisting of £35 ions or for five groups of £20
ions each. The times spent monitoring ions during sample analyses
must be the same as the times used when calibration solutions were
analyzed.
6.4 GC COLUMN — A 30 m X 0.32 mm ID fused silica capillary column coated with
a 0.25 urn or thicker film crosslinked phenyl methyl silicone (such as
Durabond-5 (DB-5), J and W Scientific, Rancho Cordova, CA) or polydiphenyl
vinyl dimethyl siloxane (such as SE-54, Alltech Associates, Deerfield, IL)
is required. Operating conditions known to produce acceptable results with
these columns are shown in Table 1; separation of pesticide analytes and PCS
calibration congeners with a DB-5 column and those operating conditions is
shown in Figure 1. Retention times have been reported (6) for all 209 PCS
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congeners with am SE-54 column/ which provides the same retention order for
PCBs and essentially the same separation capabilities as a DB-5 column.
6.5 MISCELLANEOUS EQUIPMENT
6.5.1 Volumetric flasks - 2-inL, 5-mL, 10-mL, 25-mL, and 50-mL with
ground glass stoppers.
6.5.2 Microsyringes - various standard sizes.
6.5.3 Boiling Chips — approximately 10/40 mesh. Heat at 400°C for
30 min or extract with methylene chloride in a Soxhlet apparatus.
6.5.4 Water Bath — heated, with concentric ring cover, capable of tempera-
ture control within ^ 2°C.
6.5.5 Analytical Balance — capable of accurately weighing to 0.0001 g.
7. REAGENTS AND CONSUMABLE MATERIALS
7.1 SOLVENTS — High purity, distilled-in-glass hexane and methylene chloride.
For precise injections with aplitless injectors and capillary columns, all
samples and standards should be contained in the same solvent. Effects of
. minor variations in solvent composition (i.e., small percentage of another
solvent remaining in hexane extracts) are minimized with the use of internal
standards. (External standard calibration is not acceptable.)
7.2 SODIUM SULFATE — ACS, granular, anhydrous. Purify by heating at 400°C
for 4 h in a shallow tray.
7.3 SODIUM THIOSULFATE — ACS, granular.
7.4 TETRABUTYLAMMONIUM SULFITE REAGENT — Dissove 3.39 g of tetrabutyl-
ammonium hydrogen sulfate in 100 mL distilled water. To remove impurities
extract solution three times with 20-mL portions of hexane. Discard the
hexane extracts, and add 25 g sodium sulfite to the water solution. Store
the resulting solution in an amber bottle with a Teflon-lined screw cap.
The solution can be stored at room temperature for at least one month.
7.5. MS PERFORMANCE CHECK SOLUTION — Prepare a 10 ng/uL solution of decafluoro-
triphenylphosphine (DFTPP) in an appropriate solvent.
7.6 INTERNAL STANDARDS — Chrysene-d-)2 and phenanthrene-d^o are used as internal
standards. They are added to each sample extract just before analysis and are
'contained in all calibration/performance check solutions.
7.7 SURROGATE COMPOUNDS — 13C12~4,4'-DDT and 13Cg-gamma-BHC are added to every
sample before extraction and are included in every calibration/performance
check solution.
7.8 PCS CONCENTRATION CALIBRATION CONGENERS — The nine individual PCS congeners
listed in Table 2 are used as concentration calibration compounds for PCB
determinations. One isomer at each level of chlorination is used as the
concentration calibration standard for all other isomers at that level of
chlorination, except that decachlorobiphenyl (Clig) is used for both Clg
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-8-
and C1-|Q isomer groups. The basis for selection of these calibration
congeners has been reported (7).
7.9 PCB RETENTION TIME CONGENERS FOR SIM DATA ACQUISITION OPTION — Knowledge
of the retention times of certain congeners is necessary to determine
when to acquire data with each ion set. Two concentration calibration
congeners also serve as retention time congeners; the first eluting
Cli-PCB indicates the time when data acquisition must have been initiated
for ion set #1, and the Cl^Q-PCB indicates when all PCBs have eluted.
Two or three additional PCB congeners are used to establish times to
initiate data acquisition with other ion sets (Sect. 9.4).
7.10 PESTICIDE SOLUTIONS
7.10.1 Pesticide Stock Solutions — Prepare from pure standard materials.
Weigh approximately 25.0 mg (with accuracy of 0.1 mg) of each
surrogate compound and each pure pesticide analyte, except
Endosulfan I and Endosulfan II. For those two pesticides, prepare
a stock solution twice as concentrated as that prepared for other
pesticide analytes. Dissolve each compound in hexane and dilute to
volume in a 10-mL (5-mL for the two Endosulfans) volumetric flask.
(Concentration of each component = 2.5 mg/mL, except Endosulfans,
which should be 5 mg/mL.) Smaller or larger volumes of stock solution
may be used if desired. If compound purity is certified at 2.96%,
the weight can be used without correction to calculate the concen-
tration of the stock standard solution. Commercially prepared
stock standards in hexane can be used at any concentration if they
are traceable to USEPA-supplied standards.
7.10.2 Pesticide Primary Dilution Solutions —- A convenient approach to
solution preparation is to prepare two pesticide primary dilution
solutions that are twice the concentration of the highest concentration
calibration solution required. These solutions can then be diluted
as necessary to prepare all needed calibration solutions. One solution
contains endrin aldehyde and one does not, because the medium level
calibration solution does not contain endrin aldehyde. Place 1 mL
of each pesticide analyte/surrogate compound stock solution in a
25-mL volumetric flask. (Total volume for all 22 pesticide analytes
and 2 surrogate compounds a 24 mL.) Make to volume with hexane and
mix well. (Concentration of endosulfan sufate, endosulfan I and
endosulfan II » 200 ng/uL; concentration of each other component »
100 ng/uL.)
7.11 PCB SOLUTIONS
7.11.1 Stock Solutions of PCB Calibration Congeners ~ Prepare a stock
solution of each of the nine PCB concentration calibration congeners
at a concentration of 1 ug/uL in hexane. (If SIM data are to be
acquired, prepare a 1 ug/uL stock solution of each of the three
retention time congeners also.) Place each solution in a clean
glass vial with a Teflon-lined screw cap and store at 4°C if solutions
are not to be used right away. Solutions are stable indefinitely
if solvent evaporation is prevented.
CADTION: Each time a vial containing small volumes of solutions is
warmed to room temperature and opened, a small volume of solvent in
the vial headspace evaporates, significantly affecting concentration.
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Solutions should be stored with the smallest possible volume of
headspace, and opening vials should be minimized.
7.11.2 PCS Primary Dilution Standard -- Take aliquots of the stock
solutions of the nine PCS concentration calibration congeners and
mix together in the proportions of one part of each solution of the
Clf (#1), Cl2 (#5), and Cl3 (#29) congeners/ two parts of each solution
of the C14 (#50), els (#87), and Clg (#154) congeners, three parts
of each solution-of the 017 (#188) and Clg (#200) congeners, and five
parts of the C110 (#209) congener solution. (Note: The retention
time congeners described in Sect. 7.9 are not included in the PCB
primary dilution standard because th'ey are not needed for full-range
data acquisition.) This will provide a primary dilution standard
solution of the composition shown in Table 3. Calculate the concen-
* tration in ug/uL? use three significant figures. Place each solution
in a clean glass vial with a Teflon-lined screw cap and store at
4°C. Mark the meniscus on the vial wall to monitor solution volume
during storage; solutions are stable indefinitely if solvent evapo-
ration is prevented.
7.12 INTERNAL STANDARD (IS) SOLOTIONS — Two solutions are needed to prepare
concentration calibration solutions (CALs).
7.12.1 IS.solution #1 (for full-range CALS) — Weigh 7.5 mg + 0.1 mg
each of phenanthrene-d-jg and chrysene-d^? dissolve-in hexane and
dilute to 10 mL in a volumetric flask. (Concentration of each
IS = 750 ng/uL)
7.12.2 IS solution #2 (for SIM CALS) — Take 1 mL of IS solution #1 and
dilute to 10 mL in a volumetric flask. (Concentration of each
IS - 75 ng/uL)
7.13 CALS FOR. FULL-RANGE DATA ACQUISITION — Five hexane solutions are required.
The solutions contain constant concentrations of the ISs (chrysene-d^
and phenanthrene-d^) and varying concentrations of individual pesticide
analytes, the nine PCB calibration compounds, and the two surrogate compounds
( C-2~4»4'-DDT and Ce-gamma-BHC). (Composition and approximate concen-
trations are given in Table 4.) Four solutions (high and low concentrations)
contain both ISs, both surrogate compounds, the nine PCB concentration
calibration congeners, and each of the single-compound pesticide analytes.
The fifth solution, the medium level concentration solution, contains all
the above compounds except endrin aldehyde, which is not present for reasons
described in Sect. 8. The lowest concentration solution contains each
individual pesticide analyte and each PCB calibration congener at a concen-
tration near but greater than its anticipated detection limit. (Because
MS response to PCBs decreases with increasing level of chlorination, PCB
congener concentrations in CALs increase with level of chlorination.)
Components of the highest concentration solution (High CAL) are present at
a concentration that allow injections of 2-uL aliquots without MS saturation
or GC column overloading.
7.13.1 The Full-Range High CAL can be prepared by mixing equal portions
of the PCB primary dilution solution and the pesticide primary
dilution solution that contains endrin aldehyde and then adding an
appropriate volume of IS solution #1. For example, 1 mL of each
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-10-
primary dilution solution and 20 uL of IS solution #1 provide the
appropriate concentration for High CAL.
7.13.2 Other full-range CALS are prepared by diluting the primary dilution
standard solutions and adding the appropriate amount of IS solution
#1. CADTION: The pesticide primary dilution standard that does
not contain endrin aldehyde must be used for the medium level
full-range CAL.
7.14 CALS FOR SIM DATA ACQUISITION OPTION — Two sets of solutions are needed,
one set of five solutions for determinations of pesticide analytes, and
one set of five solutions for PCB determinations. Appropriate concen-
trations of SIM CALs are given in Tables 5a and 5b. Solutions are
prepared by diluting appropriate primary dilution standards and adding
an appropriate volume of IS solution #2.
CAOTION: The Pesticide SIM Medium CAL does not contain endrin aldehyde;
the PCS SIM CALS must include the three PCB retention time congeners.
that are used to establish conditions for SIM data acquisition.
7.15 Prepare a solution of surrogate compounds in a water miscible solvent
to provide a concentration in the sample/blank extract that is near
the concentration anticipated for analytes when an aliquot of >20 uL is
added to the sample before extraction.
7.15 Calculate the concentration (two significant figures if <100 and three
significant figures if £100 ng/uL) of each component in each solution.
Note: Concentrations presented in tables are only approximate.
7.16 LABORATORY PERFORMANCE CHECK SOLUTION - For both full-range data acquisition,
and the SIM data acquisition option, the Medium CAL is used as the laboratory
performance check solution (LPC) to verify response factors and to demonstrate
adequate GC resolution and MS performance.
8- SAMPLE COLLECTION, PRESERVATION AND STORAGE
8.1 WATER SAMPLES
8.1.1 Samples must be collected in clean (Sect. 4.2) glass containers.
Note: When samples are anticipated to contain low concentrations
of method analytes, a sample larger than 1-L may be needed. An
effective sample collection procedure to minimize losses of hydro-
phobic analytes is to add a portion of extracting solvent to each
sample container when the sample is collected. When a 1-gal sample
is collected, an appropriate solvent volume is approximately 100 mL.
(The entire sample must be used as one sample aliquot, and blank
sample/solvent volumes must be adjusted also.)
8.1.2 Samples must be iced or refrigerated at 4°C from time of collection
until extraction. If samples will not be extracted within 72 h after
collection, use either sodium hydroxide or sulfuric acid to adjust
sample pH to within a range of 5 to 9. Record the volume of acid
or base used. If aldrin is to be determined, add sodium thiosulfate
when residual chlorine is present. Field test kits are available
for measurement of residual chlorine.
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8.1.3 Samples should be extracted within 7 days after collection and analyzed
within 40 days after extraction.
8.2 SOIL/SEDIMENT SAMPLES — Appropriate procedures will "be specified when
results are obtained from ongoing experiments.
9. CALIBRATION
Demonstration and documentation of acceptable initial calibration is required
before any samples are analyzed and is required intermittently throughout
sample analyses as dictated by results of continuing calibration checks.
After initial calibration is successfully performed, a continuing calibration
check is required at the beginning and end of each 12-h period during which
analyses are performed. The Medium CALs for pesticide determinations do not
include endrin aldehyde. This allows the Medium CAL to be used for continuing
calibration checks, including a check to ensure that endrin decomposition is
£10%. During initial calibration a separate Medium CAL containing endrin
aldehyde and the internal standard is analyzed to determine the response factor
for endrin aldehyde. Thereafter, if endrin aldehyde is a component of any
sample and endrin decomposition is not a problem, the response factor for
endrin aldehyde is verified by analyzing a calibration solution containing it.
9.1 DATA ACQUISITION OPTIONS — Either full-range or SIM data acquisition may
be used.
9.1.1 Full-range data acquisition is recommended if sample extract
components are anticipated to be at sufficiently high concentrations.
9.1.2 SIM data acquisition will provide an increase in sensitivity by
at least a factor of five for pesticide determinations and by at
least a factor of three for PCB determinations.
9.2. INITIAL CALIBRATION
9.2.1 Calibrate and tune the MS with standards and procedures prescribed
by the manufacturer with any necessary modifications to meet USEPA
r equi rements•
9.2.2 Inject a 1- uL or 2-uL aliquot of the 10 ng/uL DFTPP solution and
acquire a mass spectrum that includes data for m/z 45-450. If the
spectrum does not meet all criteria (Table 6), the MS must be
hardware tuned to meet all criteria before proceeding with calibration.
9.2.3 Full-Range Calibration — Inject a 1- or 2-uL aliquot of the Medium
CAL and acquire data from m/z 45 to 510. Acquire >5 spectra during
elution of each GC peak. Total cycle time should be >0.5 s and <,1*5 s.
Notes Either a 1- or 2-uL aliquot should be used consistently for
CALs and sample/blank extracts.
9.2.4 SIM Calibration -- Acquire at least five data points for each ion
during elution of each GC peak. Total cycle time should be >0.5 s
and £1«5 s.
CADTION: When acquiring SIM data, GC operating conditions must be
carefully reproduced for each analysis to provide reproducible
retention times; if not, ions will not be monitored at the appropriate
times•
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9.2.4.1 SIM Calibration for PCB determinations
9.2.4.1.1 Two options for SIM data acquisition are provided.
Data can be acquired with four sets of 60% of the
peak height of methoxychlor, which may partially coelute
with the Clg-PCB congener.
9.2.5.1.2 MS sensitivity — Signal/noise ratio of >5 for
m/z 499 of PCB congener #209, C110-PCB.
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9.2.5.1.3 MS calibration — Abundance of >40% and <60% of
m/z 502 relative to m/z 498 for PCB congener #209.
9.2.5.1.4 Lack of degradation of endrin. Examine an extracted
ion current profile (EICP) for m/z 67 in the retention
time window between 4,4'-DDE and endosulfan sulfate;
confirm that the abundance of m/z 67 at the retention
time of endrin aldehyde is <10% of the abundance of
m/z 67 produced by endrin.
9.2.5.1.5 Lack of degradation of C12-4,4'-DDT. Examine EICPs
for m/z 258 and m/z 247 in the retention time window
that includes 4,4'-DDD, 4,4'-DDE and 4,4'-DDT; m/z
258 would be produced by C12-4,4'DDE, and m/z 247 by
C.j2-4,4'-DDD. Confirm that the total abundance of
both ions is <5% of m/z 247 produced by 13C12-4,4'-DDT.
9.2.5.2 SIM PCS Data
9.2.5.2.1 GC separation -- Baseline separation of PCB congener
#87 from congeners #154 and #77, which may coelute.
9.2.5.2.2 MS sensitivity — Signal/noise ratio of £5 for m/z
499 of PCB congener #209, C110-PCB, and for m/z 241
of chrysene-d-|2-
9.2.5*2.3 MS calibration — Abundance of >70% and <95% of m/z
500 relative to m/z 498 for congener #209, C110-PCB-.
9.2.5.3 SIM Pesticide Data
9.2.5.3.1 GC separation — Baseline separation of endrin
ketone and chrysene-d12; baseline separation of
beta-BHC and gamma-BBC; baseline separation of endrin
ketone and chrysene-d12; height of chrysene-d12 peak
j>60% of methoxychlor peak height.
9.2.5.3.2 MS sensitivity — Signal/noise ratio of >5 for m/z
241 of chrysene-d-|2.
9.2.5.3.3 MS calibration — Abundance of m/z 241 relative
to that of m/z 240 produced by chrysene-di2 is >15%
and <25%.
9.2.5.3.4 Lack of degradation of endrin. Examine an SICP for
m/z 67 in the retention time window between 4,4'-DDE
and endosulfan sulfate; confirm that the abundance
of m/z 67 at the retention time of endrin aldehyde
is <10% that of m/z 67 produced by endrin.
9.2.5.3.5 Lack of degradation of 13C12-4,4'-DDT. Examine SICPs
for m/z 258 and m/z 247 in the retention time window
that includes 4,4'-ODD, 4,4'-DDE, and 4,4'-DDT; m/z
258 would be produced by C12-4,4'-DDE, and m/z 247
by 13C12-4,4'-DDD. Confirm that the total abundance
of both ions is <5% of m/z 247 produced by 13C12-4,4'-DDT.
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9.2.6 Replicate Analyses of CALs -- If all performance criteria are met,
analyze one 1- or 2-uL aliquot of each of the other four CALs.
9.2.7 Response Factor Calculation
9.2.7.1 Calculate five response factors (RFs) for each pesticide
analyte, PCB calibration congener/ and surrogate compound
relative to both ISs (See Sect. 12.3.2), phenanthrene-dig and
Ais
where AJJ * integrated ion abundance of quantitation
ion for a pesticide, a PCB calibration
congener or a surrogate compound,
integrated ion abundance of m/z 240, the '
quantitation ion when chrysene-d-j2 is used
as the internal standard or m/z 188, the
quantitation ion when phenanthrene-d-jg
is used as the internal standard,
injected quantity of chrysene-d^ or
phenanthrene-djQ,
Qx » injected quantity of pesticide analyte, PCB
calibration congener or surrogate compound.
RF is a unitless number, units used to express quantities
must be equivalent. Note: The Cl2~PCB calibration congener
may not be resolved from alpha-BHC. If not, alpha-BHC will
contribute to the ion abundance measured for C12-PCB. To
correct for this contribution, subtract 6.7% of the ion
abundance of m/z 219 measured for alpha-BHC from the ion
abundance measured for m/z 222 for
9.2.8 Response Factor Reproducibility -- For each pesticide analyte, PCB
calibration congener and surrogate compound, calculate the mean RF
from analyses of each of the five CALS. When the RSD exceeds 20%,
analyze additional aliquots of appropriate CALS to obtain an acceptable
RSD of RFs over the entire concentration range, or take action to
improve GC/MS performance.
9.2.9 SIM Analyte Retention Time Reproducibility
9.2.9.1 PCB determinations - Absolute retention times of PCB congeners
f77 and #104 should not vary by more than ;MO s from one
analysis to the next. (Retention time reproducibility is
not as critical for congeners #1 and #209 as for #77 and
#104, which are used to determine when ion sets are changed.)
9.2.9.2 Pesticide determinations — Absolute retention times of
gamma-chlordane, endosulfan I, and endosulfan II should not
vary by more than +10 s from one analysis to the next.
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9.2.10 Record a spectrum of each CAL component.
9.3. CONTINUING CALIBRATION CHECK
9.3.1 With the following procedures, verify initial calibration at the
beginning and end of each 12-h period during which analyses are to
be performed.
*
9.3.2 Calibrate and tune the MS with standards and procedures prescribed
by the manufacturer.
9.3.3 Analyze a 1-uL or 2-uL aliquot of the DFTPP solution and ensure
acceptable MS calibration and performance (Table 6).
9.3.4 Inject a 1-uL or 2-uL aliquot of the Medium CAL and analyze with the
same conditions used during Initial Calibration.
9.3.5 Demonstrate acceptable performance for criteria described in Sect.
9.2.5.
9.3.6 Determine that neither the area measured for m/z 240 for chrysene-d-) 2
nor that for m/z 188 for phenanthrene-d-) g has decreased by more than 30%
from the area measured in the most recent previous analysis of a
calibration solution or by more than 50% from the mean area measured
during initial calibration.
9.3.7 Response Factor Reproducibility — For an acceptable Continuing Cali-
bration Check, the measured RF for each analyte/surrogate compound
must be within +20% of the mean value calculated (Sect. 9.2.7)
during Initial Calibration. If not, remedial action must be taken;
recalibration may be necessary.
9.3.8 SIM Analyte Retention Time Reproducibility — Demonstrate and
document acceptable (Sect. 9.2.9) reproducibility of absolute retention
times of appropriate pesticide analytes and PCS retention time congeners.
9.3.9 Remedial actions must be taken if criteria are not met; possible
remedies are:
9.3.9.1 Check and adjust GC and/or MS operating conditions.
9.3.9.2 Clean or replace injector liner.
9.3.9.3 Flush column with solvent according to manufacturers
instructions.
9.3.9.4 Break off a short portion (approximately 0.33 m) of the
column; check column performance by analysis of performance
check solution.
9.3.9.5 Replace GC column; performance of all initial calibration
procedures then required.
9.3.9.6 Adjust MS for greater or lesser resolution.
9.3.9.7 Calibrate MS mass scale.
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9.3.9.8 Prepare and analyze new concentration calibration/
performance check solution.
9.3.9.9 Prepare new concentration calibration "curve(s).
10. QUALITY CONTROL
10.1 LABORATORY REAGENT BLANK (LRB) — Perform all steps in the analytical
procedure (Section 11)'using all reagents, standards/ surrogate compounds,
equipment, apparatus, glassware, and solvents that would be used for a
sample analysis, but omit an aliquot of sample (water or soil/sediment).
For water samples, substitute 1 L of reagent water. If available,
substitute EPA-provided reagent blank solid material for an aliquot of
soil/sediment.
10.1.1 An LRB must contain the same amount of surrogate compounds and
internal standards that is added to each sample. This amount
will vary with sample type and with the type of data acquisition
(full-range or SIM).
10.1.2 Analyze an LRB before any samples are extracted and analyzed.
10.1.3 Before a new batch of solvents or reagents is used for sample
extraction or for'column chroraatographic procedures, analyze
'an LRB. In addition, analyze a laboratory solvent blank (LSB),
which is the same as -an LRB except that no surrogate compounds or
internal standards are added; this demonstrates that reagents
contain no impurities producing an ion current above the level of
background noise for quantitation ions for those compounds.
10.1.4 Analyze an LRB along with each batch of £20 samples.
10.1.5 An acceptable LRB contains no method analyte at a concentration
greater than one half of its MDL and contains no additional compounds
with elution characteristics and mass spectral features that would
interfere with identification and measurement of a method analyte
at its MDL. If the LRB that was extracted along with a batch of
samples is contaminated, the entire batch of samples must be
reextracted and reanalyzed.
10.1.6 Corrective action for unacceptable LRB ~ Check solvents, reagents,
apparatus and glassware to locate and eliminate the source of
contamination before any samples are extracted and analyzed.
Purify or discard contaminated reagents and solvents.
10.2 CALIBRATION — Included among initial and continuing calibration procedures
are numerous quality control checks to ensure that valid data are acquired
(See Sect. 9). Continuing calibration checks are accomplished with results
from analysis of one solution, the medium level calibration solution for
the appropriate type of data acquisition, either full-range or SIM.
10.2.1 If some criteria are not met for a Continuing Calibration Check
after a 12-h period during which samples were analyzed, those
samples must be reanalyzed. Those criteria are: GC performance
(Sect. 9.2.5), MS calibration as indicated by DFTPP spectrum, and
MS sensitivity as indicated by area of internal standards.
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10.2.2 When other criteria in Sect. 9.2 are not met, results for affected
analytes must be labeled as suspect to alert the data user of the
observed problem. Included among those criteria are: response
factor check for each analyte or PCS calibration congener, degra-
dation of DDT and endrin, and retention time reproducibility for
SIM data acquisition.
10.3 INITIAL DEMONSTRATION OF LABORATORY CAPABILITY FOR WATER ANALYSES
(Insufficient information is currently available for demonstration for
soil/ sediment analyses.)
10.3.1 Until appropriate Quality Control Check Samples are available,
each laboratory should prepare one or more solutions containing
each method analyte at a concentration corresponding to that antici-
pated in samples. Until accuracy and precision limits have been
established for PCS isomer groups in appropriate samples, a solution
containing an Aroclor mixture may be used; compare total measured
PCB concentration to the total Aroclor concentration. Report
Aroclor concentration and measured concentrations of PCS isomer
groups and total measured PCS concentration.
10.3.2 Add an appropriate volume of a solution of method analytes
to each of four 1-L aliquots of reagent water. Extract and
analyze according to procedures in Sect. 11.
10.3.2 For each analyte, calculate measured concentrations, relative
standard deviation of the four measurements, and method bias
(Sect. 12.6). .
tO.4 LABORATORY PERFORMANCE CHECK SOLUTION — In this method, the medium level
concentration calibration solution also serves the purpose of a laboratory
performance check solution.
10.5 LABORATORY SURROGATE SPIKE
10.5.1 Measure the concentration of both surrogate compounds in
every sample and blank.
10.5.2 Until performance based acceptance limits have been established for
surrogate compounds, the following guidelines are provided:
measured bias with LRB =• -30% to +10%; measured bias with
water or soil/sediment extract » -50% to +25%.
10.6 QUALITY CONTROL CHECK SAMPLE — Not yet available; anticipate need for
analysis of one for each batch of £20 samples. If full-range data are
acquired, both pesticide and PCS analytes can be determined with one
analysis. If SIM data are acquired, one extraction and two GC/MS analyses
will be needed to determine both PCBs and pesticides.
10.7 LABORATORY SPIKED DUPLICATE SAMPLE — Select one sample from each batch of
£20 samples of similar type and fortify (spike) two aliquots of that sample
with a solution containing appropriate concentrations of pesticide analytes
and at least one Aroclor mixture. After addition of surrogate compounds,
extract and analyze (Sect. 11) these two fortified aliquots along with
an additional unfortified sample aliquot. Relative difference (RD) of
duplicate results for surrogate compound concentrations should be £40%.
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(RD - [C-! - C2 / 0.5 (CT + C2)] 100 ) Calculate bias (Sect. 12.6) for
each analyte and surrogate compound. Insufficient data are currently
available to provide guidance for acceptable bias and RD of -measured
analyte concentrations.
10.8 PERFORMANCE EVALUATION SAMPLE — Not yet available; to be analyzed
periodically when available.
11. PROCEDURES
11.1 SAMPLE EXTRACTION
11.1.1 Water Samples
11.1.1.1 Mark the water meniscus on the side of the sample bottle
for later determination of sample volume. Pour entire
sample into a 2-L separatory funnel. (If a sample larger
than 1-L or 1-
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and the water temperature as required to complete the
concentration in 15-20 min. At the proper rate of
distillation the balls of the column will actively chatter
but the chambers will not flood with condensed solvent.
When the apparent volume of liquid reaches 1 mL, remove
the K-D apparatus from the water bath and allow it to
drain and cool for at least 10 min.
11.1.1.6 Momentarily remove the Snyder column, add 50 mL of hexane
and a new boiling chip, and reattach the Snyder column.
Increase the temperature of the hot water bath to about
80 °C. Concentrate the extract to approximately 10 mL
as in Sect. 11.1.1.5, except use hexane to prewet the
column. Elapsed time of concentration should be 5-10
min.
11.1.1.7 Remove the Snyder column and rinse the flask and its
lower joint into the concentrator tube with 1-2 mL of
hexane. 'A 5-mL syringe is recommended for this operation.
Stopper the concentrator tube and store refrigerated if
further processing will not be performed within a few
hours. If the extract will be stored longer than two
days, transfer it to a Teflon-sealed screw-cap vial.
11.1.1.8 Determine the original sample volume by refilling the
sample bottle to the mark and transferring the liquid
to a 1000-mL graduated cylinder. Record the sample
volume to the nearest 5 mL.
11.1.2 Soil/Sediment Samples — Appropriate extraction procedures to be
specified when results of ongoing experiments are obtained.
11.2 Sulfur Removal — Elemental sulfur can be removed by the procedure described
below. (Sulfur is not expected to be a problem in water sample extracts but
sulfur removal is recommended for soil/sediment sample extracts.)
11.2.1 Transfer the extract to a 50-mL clear glass bottle or vial with a
Teflon-lined screw cap. Rinse the extract container wtih 1.0 mL of
hexane, adding the rinse to the 50-mL bottle.
11.2.2 Add 1 mL of Tetrabutylammonium-sulfite reagent and 1 mL 2-propanol,
cap the bottle, and shake for at least 1 min. If the sample is
colorless or if the initial color is unchanged, and if clear crystals
(precipitated sodium sulfite) are observed, sufficient sodium
sulfite is present. If the precipitated sodium sulfite disappears,
add more crystalline sodium sulfite in approximately 100-mg portions
until a solid residue remains after repeated shaking.
11.2.3 Add 5 mL of distilled water and shake for at least 1 min. Allow
the sample to stand for 5-10 min and remove the hexane layer (top)
for analysis. Dry the extract by passing it through a 10-cm
column containing hexane-washed sodium sulfate. Rinse the sodium
sulfate with about 30 mL of hexane and add this hexane to the
extract. Concentrate the extract to approximately 10 mL with a
K-D apparatus. Store in a refrigerator if GC/MS analysis is not to
be performed within a few hours.
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11.3 GC/MS ANALYSIS
11.3.1 Remove the sample extract or blank from storage and allow it to warm
to ambient laboratory temperature if necessary. With a stream of
dry, filtered nitrogen, reduce the extract/blank volume to the
appropriate volume, depending on anticipated analyte concentrations.
Add an appropriate volume of the appropriate internal standard stock
solution.
11.3.1.1 Internal standard concentration for full-range
data acquisition =« 7.5 ng/uL of extract.
11.3*1.2 Internal standard concentration for SIM data
acquisition - 0.75 ng/uL of extract.
11.3.2 Inject a 1-uL or 2-uL aliquot of the blank/sample extract into the GC
operated under conditions used to produce acceptable results during
calibration.'
11.3.3 Acquire mass spectral data with either full-range data acquisition
conditions or SIM conditions, as appropriate. Use the same data
acquisition time and MS operating conditions previously used to
determine response factors.
11.3.4 Examine data for saturated ions in mass spectra of target compounds,
if saturation occurred, dilute and reanalyze the extract after the
quantity of the internal standards is adjusted appropriately.
11.3.5 For each internal standard, determine that the area measured in the
sample extract has not decreased by >30% from the area measured
during the most recent previous analysis of a calibration solution
or by >50% from the mean area measured during initial calibration.
If either criterion is not met, remedial action must be taken to
improve sensitivity, and the sample extract must 'be reanalyzed.
11.4 IDENTIFICATION PROCEDURES
11.4.1 Using the ions shown in Tables 7a-7c for PCBs or Table 9 for
pesticides, examine ion current profiles (ICPs) to locate internal
standards, surrogate compounds, pesticide analytes, and PCBs for each
isomer group. Use the RRT data in Table 9 as guidelines for location
of pesticide analytes and the RRT window data in Table 3 as guidelines
for location of PCS isomers. (A reverse search software routine
can be used to locate compounds of concern.)
11.4.2 Full-Range Data
11.4.2.1 Examine.each pesticide and PCS candidate spectrum after
background correction routines have been applied. Compare
the candidate spectrum with the appropriate standard spectrum
measured during calibration. Verify the absence of any ions
with mass greater than the highest mass possible for the
compound of concern. (Ions in PCS M+ ion clusters are shown
in Table 12.)
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11.4.2.2 Obtain integrated abundance areas for quantitation and
confirmation ions.
11.4.3 SIM Data — Obtain appropriate selected ion current profiles (SICPs)
for IS quantitation and confirmation ions, for each ion monitored
to detect pesticides and the surrogate compounds (Table 9), and for
the quantitation and confirmation ions for each PCB isomer group.
11.4.4 PCB Analytes
11.4.4.1 For all PCB candidates, confirm the presence of an (M-?!))*
ion cluster by examining ICPs or spectra for at least one of
the most intense ions in the appropriate ion cluster.
11.4.4.2 For Cl^-Cl7 isomer groups, examine ICPs or spectra for intense
(M+70)* ions that would indicate a coeluting PCB containing two
additional chlorines. (GC retention time data show that
this is not a potential problem for other PCB isomer groups;
see Figure 2.) If this interference occurs, a correction can
be made. Obtain and record the area for the appropriate ion
(Table 12) for the candidate PCB isomer group. Use the
information in Table 13 to correct the measured abundance of
M . For example, if a Cl^-PCB and a Clg-PCB candidate coelute,
the Cly-PCB will contribute to the ion measured for m/z 326 and
m/z 324, the quantitation and confirmation ions, respectively,
for a C15-PCB. Obtain and record the area for m/z 322 (the
lowest mass ion in the (M+-70) ion cluster of a Cl^-PCB
fragment produced by a Cl7-PCB)t. To determine the m/z 326 and
m/z 324 areas produced by the Cls PCB, calculate the C17-PCB
contribution to each and subtract it from the measured area.
In this example, 164% of the area measured for m/z 322 should
be subtracted from the area measured for m/z 324, and 108% of
the m/z 322 area should be subtracted from the area measured
for m/z 326 (Table 13).
11.4.4.3 For Cl2-Clg-PCB candidates, examine ICPs or spectra for
intense (M+35)+ ions that would indicate a coeluting PCB
containing one additional chlorine. This coelution causes
interferences because of the natural abundance of 13C.
(This interference will be small and can be neglected except
when measuring the area of a small amount of a PCB coeluting
with a large amount of another PCB containing one more
chlorine.) To correct for this interference, obtain and
record the area for the appropriate ion (Table 14) from
the (M-1)+ ion cluster, and subtract 13.5% of the area
measured for the CM-1)"*" ion from the measured area of the
quantitation ion. For example, for Cls-PCB candidates,
obtain and record the area for m/z 325; subtract 13.5% of
that area from the measured area of m/z 326.
11.4.5 All Analytes — Use ICP data to calculate the ratio of the measured
peak areas of the quantitation ion and confirmation ion(s), and
compare to the acceptable ratio (Table 9 for pesticides and Table 12
for PCBs). If acceptable ratios are not obtained, a coeluting or
partially coeluting compound may be interfering. Examination of data
from several scans may provide information that will allow application
of additional background corrections to improve the ion ratio.
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11.5. IDENTIFICATION CRITERIA
11.5.1 Internal Standards
11.5.1.1 Chrysene-d12 — the abundance of m/z 241 relative to m/z
240 must be XI5% and £25%, and these ions must maximize
simultaneously. The area measured for m/z 240 must be
within 30% of the area measured during the most recent
calibration.
11.5.1.2 Phenanthrene-d-jQ ~ the abundance of m/z 189 relative to m/z
188 must be _>10% and £22%, and these ions must maximize
simultaneously. The area measured for m/z 188 must be
within 30% of the area measured during the most recent
acceptable calibration.
11.5.1.3 Retention time must be within £10 s of that observed
during the most recent acceptable calibration.
11.5.2 Full-Range Data for Pesticide Analytes and Surrogate Compounds
11.5.2.1 Retention time of the sample component must be within t s
of the time observed for that same compound when a calibration
solution was analyzed. Calculate the value of t with the
equation, t = (RT)V^» where RT = observed retention time
(in seconds) of the compound during the last previous acceptable
calibration. ' .
11.5.2.2 All ions with relative abundance >10% in the mass spectrum
must be present in the mass spectrum of the candidate sample
component; a molecular ion with relative abundance >2% in
the standard spectrum must be present in the candidate
spectrum.
11.5.2.3 The ion that was the most abundant (base peak) in the standard
spectrum must also be the base peak in the candidate spectrum.
11.5.2.4 For all ions with relative abundance >20% in the standard
spectrum, the relative abundance in the candidate spectrum
must not vary by more than £15% in percentage units (i.e.,
if 50% in standard, must be £35% and £65%).
11.5.2.5 Ions with relative abundance >10% in the candidate spectrum
but not present in the standard spectrum must be considered
and accounted for by the analyst. When data processing
software is used to obtain candidate spectra, both processed
and unprocessed spectra must be evaluated.
11.5.3 SIM Data for Pesticide Analytes and Surrogate Compounds
11.5.3.1 Absolute retention time of each surrogate compound and
pesticide candidate must be within 10 s of that measured
during the last previous acceptable calibration.
11.5.3.2 All ions monitored for each compound (Table 9) must be
present and must maximize simultaneously.
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11.5.3.3 In a spectrum averaged across a GC peak and with background
correction, if necessary/ the most abundant ion must correlate
with Table 9 data.
11.5.3.4 Observed relative abundances of the monitored ions must
meet the following criteria:
Aldrin — m/z 263 =- >20% and m/z 265 = >13%
BBC (each isomer) — m/z 183 » 70-95% of m/z 181
13C6-gamma-BHC — m/z 189 = 75-90% of m/z 187
Chlordane (alpha and gamma) — m/z 375 » 75-99%
4,4'-DDE — m/z 248 = 45-85%
4,4'-ODD and 4,4'-DDT — m/z 237 - 45-85%
13C12-4,4'-DDT — m/z 249 - 45-85%
Dieldrin — m/z 263 = >3% and m/z 108 » >8%
Endosulfan I and II ~ m/z 339 = >30% and m/z 341 =• >20%
Endosulfan sulfate — m/z 274 = 60-95%
' Endrin — m/z 263 = >50%
Endrin aldehyde —• m/z 345 = _>m
Endrin ketone — m/z 317 = 2.30%
Heptachlor — m/z 272 » >30% and m/z 274 = >20%
Heptachlor epoxide ~ m/z 353 = >60%
Methoxychlor — m/z 228 « 3-30%
Nonachlor — m/z 407 = 65-95%
11.5.4 Full-Range and SIM Data for PCBs
11.5.4.1 Absolute retention times of surrogate compounds must be
within vt 0 s of that measured during the last previous
continuing calibration check.
11.5.4.2 Quantitation and confirmation ions for each PCB isomer group
must maximize within ^1 scan of each other.
11.5.4.3 The integrated ion current for each quantitation and confir-
mation ion must be at least three times background noise and
must not have saturated the detector.
11.5.4.4 For each PCB isomer group candidate, the ratio of the quanti-
tation ion area to the confirmation ion area must be within
limits shown in Table 12; at least one ion in the (M-70)+
ion cluster must be present.
12. CaLCOIATIONS
12.1 From appropriate ICPs of quantitation ions, obtain and record the spectrum
number of the chromatographic peak apex and the area of the entire
chromatographic peak.
12.2 For PCBs, sum the areas for all isomers identified at each level of
chlorination (e.g., sum all quantitation ion areas for Cl^PCBs).
12.3 Calculate the concentration of each surrogate compound, pesticide
candidate, and PCB isomer group using the formula:
-------
-24-
Cx - /(Ais '«*•»)
where Cx = concentration (micrograms per kilogram or micrograms
per liter) of surrogate compound, individual pesticide
or a PCB isomer group,
AX = the area of the quantitation ion for each pesticide
analyte/surrogate compound or the sum of quantitation
ion areas for all PCB isomers at a particular level
of chlorination,
the area of the internal standard quantitation ion,
m/z 240 for chrysene-d^ °r m/z 188 for phenanthrene-d^ g,
quantity (micrograms) of internal standard added to
the extract before GC/MS analysis,
RF = calculated response factor for the surrogate compound,
the pesticide standard, or the PCB calibration compound
for the isomer group (level of chlorination), and
W =• weight (kilograms) of sample extracted. If a liquid
sample was extracted, W becomes V, the volume (liters)
of water extracted, and concentration units become
micrograms per liter.
12.3.1 Use the grand mean RF calculated during Initial Calibration.
CAOTION: For PCB analyses with automated data interpretation
a linear fit algorithm will produce erroneous concentration
data.
12.3.2 For pesticides eluting before heptachlor epoxide, use the RF
relative to phenanthrene-d-jg; for heptachlor expoxide and later
eluting pesticides, use the RF relative to chrysene-d-)2' For
PCBs, use the RF relative to chrysene-d^ unless an interference
makes the use of the RF relative to phenanthrene-d-jg necessary.
12.4 Estimation of the Concentration of Technical Chlordane. Technical chlordane
is a mixture that contains alpha-chlordane (about 13% by weight), gamma-
chlordane (about 18% by weight), heptachlor (about 8%), chlordene (three
isomers; about 19%) and a variety of side reaction products (including
nonachlor isomers) from chlorination of chlordene. Alpha-chlordane is
readily converted to gamma-chlordane, which is persistent in environmental
samples. Another persistent component is trans-nonachlor. The presence
of gamma-chlordane and trans-nonachlor, with or without alpha-chlordane
and heptachlor) indicates that technical chlordane was once present in
the sample. Therefore the sum of measured concentrations of alpha-chlordane
and gamma-chlordane can be used to estimate the original concentration of
technical chlordane.
ctc =
-------
-25-
12.5 Report calculated values to two significant figures.
12.6 When samples of known composition or fortified samples are analyzed,
calculate the percent method bias using the equation:"
B - 100 (Cs - Ct)/ Ct
where Cg ™ measured concentration (in micrograms per kilogram
or micrograms per liter),
Ct » theoretical concentration (i.e., the
quantity added to the sample aliquot/weight or volume
of sample aliquot)•
Note: The bias value retains a positive or negative sign.
*
13. AUTOMATED IDENTIFICATION AND MEASUREMENT
Special software can be used for automated identification and measurement of
PCBs (8) and pesticides. Unprocessed GC/MS data are handled without human
interaction with the software operating on the dedicated computer. A concen-
tration for each pesticide and each PCB isomer group is calculated automatically.
Contact EMSL-Cincinnati for further information.
14. METHOD PERFORMANCE
To obtain single laboratory accuracy and precision data for method analytes,
replicate 1-L aliquots of reagent water and river water fortified with known
amounts of analytes were extracted and analyzed. Automated procedures were used
to identify and measure method analytes in 2-uL aliquots of 1-mL extracts.
Because a sufficient quantity of individual PCB congeners was not available,
Aroclor mixtures were used to fortify water samples. This is not desirable,
because individual PCBs in Aroclors vary in concentration. As Aroclor concen-
trations decrease in a sample extract, an increasing number of components
will fall below the detection limit and will not be identified and measured.
In addition, insufficient data are available about Aroclor composition to assess
accuracy of isomer group measurements or to assess MDLs for PCBs when Aroclors
are used to fortify samples.
14.1 Medium Level Reagent Water Extracts —• Five aliquots of reagent water
fortified with each individual pesticide at a concentration of 10 ug/L and
Aroclors 1221, 1242, 1254, and 1268 at concentrations of 5 ug/L, 50 ug/L,
50"ug/L and 25 ug/L, respectively, were extracted and analyzed. Method
bias for individual pesticides ranged from -10% to +18% with a mean method
bias of +2% for all 21 pesticides (Table 15). For individual pesticides,
RSDs of measured concentration ranged from 0.61% for endrin ketone to
9.8% for endrin aldehyde. No true values are known for concentrations of
PCB isomer groups in Aroclors, but the mean measured total PCB concentration
was 110 ug/L (RSD 2.9%), which indicated a method bias of -15%. For
individual isomer groups, RSDs of mean measured concentrations ranged
from 3.9% to 16%.
14.2 Low Level Reagent Water Extract -- Reagent water was fortified with each
pesticide at a concentration of 3 ug/L and a total PCB concentration of
27 ug/L (Aroclors 1221, 1 ug/L; 1242, 10 ug/L; 1254, 10 ug/L; and 1268,
-------
-26-
6 ug/L). When seven replicate extracts were analyzed, method bias for
individual pesticides ranged from -17% to +20% with a mean method bias of
-2% (Table 15). An MDL was calculated for each pesticide using the equation
relating the standard deviation of the seven replicate measurement and
Student's t value for a one-tailed test at the 99% confidence level with n-1
degrees of freedom (1). With this calculation, MDL is defined as the
minimum concentration that can be measured and reported with 99% confidence
that the value is above zero. The excellent precision achieved with these
measurements resulted in unrealistically low MOLs ranging from 0.2 to 0.8
ug/L for pesticide analytes (Table 15). A PCS MDL is an individual congener
characteristic and cannot be determined with samples fortif-ied with Aroclor
mixtures. Estimates of MDLs for individual components of PCS isomer groups
were obtained by proportioning the total quantity measured for each isomer
group among individual measured isomers. The estimated MDL values for
individual PCBs also were unrealistically low (0.01-0.1 ug/L) because of
the excellent precision of measurements. A more realistic statement of
detection limits for pesticides and PCBs can be found in Sect. 1.2.
14.3 River Water Extracts ~ Five aliquots of river water fortified with
each pesticide at a concentration of 5 ug/L and total PCB concentration
of 70 ug/L (Aroclors 1221, 2 ug/L; 1242, 30 ug/L; 1254, 30 ug/L; and
1268, 8 ug/L) were extracted and analyzed. Method bias for individual
pesticides ranged from -30% to +8% with a mean of -8% (Table 15). The
excellent precision of measured pesticide PCS isomer group concentrations
was indicated by RSDs ranging from 1.6% to 7.5%. The mean measured total
PCS concentration of 51 ug/L (RSD 2.5%) indicated a method bias of -27%.
15. REFERENCES
1. Glaser, J. A., D. L. Foerst, G. D. McKee, S. A. Quave, and W. L. Budde,
"Trace Analyses for Wastewaters", Environ. Sci. Technol. 15, 1426, 1981.
2. Ballschmiter, K. and M. Zell, Fresenius Z. Anal. Ghent., 302, 20, 1980.
3. "Carcinogens — Working with Carcinogens", Department of Health Service,
Center for Disease Control, National Institute for Occupational Safety
and Health, Publication No. 77-206, August 1977.
4. "OSHA Safety and Health Standards, General Industry", 29 CFR 1910,
Occupational Safety and Health Administration, OSHA 2206, Revised
January 1976.
5. "Safety in Academic Chemistry Laboratories", American Chemical Society
Publication, Committee on Chemical Safety, 3rd Edition, 1979.
6. Mullin, M. D., C. Pochini, S. McCrindle, M. Romkes, S. H. Safe, and
L. M. Safe, "High Resolution PCS Analysis: Synthesis and Chromatographic
Properties of All 209 PCS Congeners", Environ. Sci. Technol. 18, 466, 1984.
7. Gebhart, J. E., Hayes, T. L., Alford-Stevens, A. L., and W. L. Budde,
"Mass Spectrometric Determination of Polychlorinated Biphenyls as
Isomer Groups", Anal. Ghent. 57, 2458, 1985.
-------
-27-
8. Slivon, L. E., J. E. Gebhart, T. L. Hayes, A. L. Alford-Stevens/
W. L.-Budde, "Automated Procedures for Mass Spectrometric Determi-
nation of Polychlorinated Biphenyls as Isomer Groups", Anal. Chem.
£7, 2464, 1985.
9. . Rote, .&• W. and W. J. Morris, "Use of Isotopic Abundance Ratios in
Identification of Polychlorinated Biphenyls by Mass Spectrometry",
J. Assoc. Offic. Anal." Chem. 56(1), 188, 1973.
Table 1. Recommended GC Operating Conditions
Column Type:
Film Thickness:
Column Dimensions:
Helium Linear Velocity:
Temperature Program for Splitless Injection:
o Full-range data acquisition for PCBs
and pesticides
(Analysis time = approx. 50 min)
o SIM data acquisition for PCBs
(Analysis time = approx. 25 min)
o SIM data acquisition for pesticides
(Analysis time - approx. 30 min)
SE-54 or DB-5
0.25 urn
30 m X 0.32 mm
28-29 cm/sec
at 250°C
Inject at 80°C and hold 1 min;
increase at 30°/min to 160°C and
hold 1 min? increase at 3°/min to
310«C.
or
Inject at 80°C and hold 1 min; heat
rapidly to 160°C and hold 1 min;
increase at 3°/min to 310°C.
Inject at 45°C and hold 1 min; increase
at 20°/min to 150«C and hold 1 min;
increase at 10°/min to 310°C.
Inject at 80 °C and hold 1 min; increase
at 30°/min to 160°C and hold 1 min;
increase at 3°/min to 250°C; hold
past elution time of methoxychlor.
-------
-28-
Table 2. PCS Congeners Used as Calibration Standards
Congener Chlorine
PCS Isomer Group Number3 Substitution
Concentration Calibration Standard
Monochlorobiphenyl 1 2
Dichlorobiphenyl 5 2,3
Trichlorobiphenyl 29 2,4,5
Tetrachlorobiphenyl 50 2,2',4,6
. Pentachlorobiphenyl 87 2,2',3,4,5'
Hexachlorobiphenyl 154 2,2',4,4',5,6'
Hepta-chlorobiphenyl 188 2,2',3,4',5,6,6'
Octachlorobiphenyl 200 2,2'',3,3',4,5',6,6'
Nonachlorobiphenyl*5 - -—
Decachlorobiphenyl 209 2,2',3,3',4,4',5,5',6,6'
Retention Time Calibration Standards
Tetrachlorobiphenyl 77 3,3',4,4'
Pentachlorobiphenyl 104 2,2',4,6,6'
Nonachlorobiphenyl 208 2,2',3,3',4,5,5',6,7'
a Numbered according to the system of Ballschmiter and Zell (2).
b Decachlorobiphenyl is used as the calibration congener for both nona-
and decachlorobiphenyl isomer groups.
-------
-29-
Table 3. Scheme for Preparation of PCB Primary Dilution Standard
PCB
Cong.
#1
#5
#29
#50
#87
#154
#188
#200
#209
Isomer
Group
Cl,
C12
C13
C14
C15
<*6
C17
Clg
d10
Stock Sol.
Cone.
mg/mL
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
Proportion
for Primary
Oil. Sol.
1 part
1 part
1 part
2 parts
2 parts
2 parts
3 parts
3 parts
5 parts
Primary Oil.
Std. Cone.
ng/uL
50
50
50
100
100
100
150
150
250
Total 20 parts
-------
-30-
Table 4. Composition and Approximate Concentrations of Calibration Solutions
for Pull-Range Data Acquisition
Analyte/Int. Std./
Surrogate Compound
CAL 1
Concentration (ng/uL)
CAL 2 CAL 3 CAL 4 CAL 5
PCS Cal. Congeners
Cl! (#1)
C12 (#5)
C13 (#29)
C14 (#50)
C15 (#87)
C16 (#154)
C17 (#188)
C18 (#200)
C110 (#209)
Pesticides
Aldrin
BHC, each isomer
Chlordane, each isomer
4, 4 '-ODD
4, 4 '-DDE
4, 4 '-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Heptachlor
Heptachlor epoxide
Methoxychlor
Nonachlor, each isomer
Internal Standards
Phenanthrene-d-j Q
Surrogate Compounds
13C6-gamma BHC
0.5
0.5
0.5
1
1
1
1.5
1.5
2.5
2.5
2.5
2.5
5
5
5
7.5
7.5
12.5
5
5
5
10
10
10
15
15
25
10
10
10
20
20
20
30
30
50
25
25
25
50
50
50
75
75
125
1
1
1
1
1
1
1
2
2
2
1
1
1
1
1
1
1
5
5
5.
5
5-.
5
5
10
10
10
5
5
5
5
5
5
5
10
10
10
10
10
10
10
20
20
20
10
—
10
10
10
10
10
20
20
20
20
20
20
20
40
40
40
20,
20
20
20
20
20
20
50
50
50
50
50
50
50
100
100
100
50
50
50
50
50
50
50
13
C12-4,4'-DDT
7.5
7.5
7.5
7.5
5
5
7.5
7.5
10
10
7.5
7.5
20
20
7.5
7.5
50
50
-------
-31-
Table 5a. Composition and Approximate Concentrations of Calibration Solutions
for SIM Data Acquisition for PCS Determinations
Concentration (ng/uL)
Compound
Cal. Congeners
C1-, (#1)
C12 (#5)
C13 (#29)
C14 (#50)
C15 (#87)
C16 (#154)
C17 (#188)
Clg (#200)
C110 (#209)
RT Congeners
C14 (#77)
C15 (#104)
C19 (#208)
Internal Standards
Chrysene-di 2
Phenanthr ene-df g
CAL 1 CAL 2
0.1 0.5
0.1 0.5
0.1 0.5
0.2 1.0
0.2 1
0.2 1
0.3 1.5
0.3 1.5
0.5 2.5
0.2 1
0.2 1
0.4 2
0.75 0.75
0.75 0.75
CAL 3
1
1
1
2
2
2
3
3
5
2
2
4
0.75
0.75
CAL 4
2
2
2
4
4
4
6
6
10
4
4
8
0.75
0.75
CAL 5
5
5
' 5
10
10
10
15
15
25
10
10
20
0.75
0.75
Surrogate Compounds
^Cg-gamma-BHC
13
C12-4,4'-DDT
0.2
0.2
2
2
4
4
10
10
-------
-32-
Table 5b. Composition and Approximate Concentrations of Calibration Solutions
for SIM Data Acquisition for Pesticide Determinations
Concentration (ng/uL)
Analyte/Internal Std/
Surrogate Compound
Pesticide Analytes
Aldrin
BHC, each isomer
Chlordane, each isomer
4, 4 '-ODD
4, 4 '-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Heptachlor
Heptachlor epoxide
Methoxychlor
Nonachlor, each isomer
Internal Standards
Chrysene-d-|2
Phenanthrene-di g
Surrogate Compounds
Cg-gamma-BHC
13C-,-4,4'-DOT
CAL 1
0.2
0.2
0.2
0.2
0.2
0.2
0.2
0.4
0.4
0.2
0.2
0.2
0.2
0.2
0.2
0.2
0.2
0.75
0.75
0.2
0.2
CAL 2
1
1
1
1
1
1
1
2
2 '
1
1
1
1
1
1
1
1
0.75
0.75
1
1
CAL 3
2
2
2
2
2
2
2
4
4
2
2
-
2
2
2
2
2
0.75
0.75
2
2
CAL 4
5
5
5
5
5
5
5
'10
10
5
5
5
5
5
5
5
5
0.75
0.75
5
5
CAL 5
10
10
10
10
10
10
10
20
20
10
10
10
10
10
10
10
10
0.75
0.75
10
10
-------
-33-
Table 6. Criteria for DFTPP Spectrum
m/z " Relative Abundance
127 40-60%
197 <1%
198 100% (Base Peak)
199 5-9%
275 10-30%
365 >1%
441 ' Present and 40%
443 . 17-23% of m/z 442
-------
-34-
Table 7a. Ions for Selected Ion Monitoring to Determine PCBs by Acquiring
Data for Four Sets of <35 Ions Each
PCS Isomer Group/
Int.Std./Surr.Cmpd.
Monochlorobiphenyls
Dichlorobiphenyls
Trichlorobipheny Is
Tetrachlorobiphenyls
Pentachlorobiphenyls
Hexachlorobiphenyls
Hept a chlor obipheny Is
*
Octachlorobiphenyls
No nach 1 or obi pheny Is
Decachlorobiphenyl
Chrysene-d-j 2
Phenanthr ene-d^ g
1 3Cg-gamma-BHC
13C1--4/4'-DOT
Nominal
Mol. Wt.
188
222
256
290
324
358
392
426
460
494
240
188
294
364
Mass or Range
to be Monitored
152; 186-190
220-224
254-260
288-294
322-328
356-362
390-396
424-430
460-466
496-500
240-241
188-189
187,189
247; 249
No. of
Ions
6
5
7
7
7
7
7
7
7
5
2
2
2
2
Ion Sets
#1 #2 #3 #4
6
5
7 7 1a
7 7 1b
7 7
6C 7 7
6d 7
7
7
5
2
2e
2f
2
Total # ions 25 27 24 35
Monitor m/z 254 to confirm presence of (M-70) for Clg-PCBs.
Monitor m/z 288 to confirm presence of (M-70)+ for Clg-PCBs..
GBegin range at m/z 357 in Ion Set #2.
^Begin range at m/z 391 in Ion Set #3.
eM/z 188 and 189 included among ions used to detect and measure monochlorobiphenyIs<
187 and 189 included among ions used to detect and measure monochlorobiphenyls.
-------
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CM CM P) P)
r- CM P)
r-l r* i-4
CJ CJ
>O O r» r>
CM vo cn f
P^ P> P) CM
§
a
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vo cn PI ^*
PI PI ^f CM
CM
o •*• oo
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ro T»" in vo
^ rH r-l r-l
CJ CJ CJ CJ
1
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rH r-( H TJ>
CJ CJ CJ 1
CM
u~
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4)
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rH rH rH QQ
CJ U O >i
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CJ
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co cn «-
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CM
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D i-t
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-------
-36-
Table 7c. Ions for Selected Ion Monitoring to Determine PCBs by Acquiring
Data for Five Ion Sets of <20 Ions
Ion Set Ion Set Ion Set Ion Set Ion Set
No. 1a No. 2b No. 3C No. 4d No. 5e
"152 186 - 247 240 356
153 188 249 241 358
186 220 254 288 360
187 222 256 290 390
188 254 288 322 392
189 255 290 324 394
190 256 322 326 424
220 258 323 356 425
221 288 324 357 426
222 289 * 326 358 428
224 290 328 360 430
255 292 357 362 432
256 294 358 391 462
258 323 360 392 464
290 324 362 394 466
292 326 392 396 496
294 . 328 394 398 498
358 396 428 499
360 398 430 500
362 . 432 502
17 ions 20 ions 19 ions 20 ions 20 ions
a Ions to identify and measure Cl..-Cly-PCBs, phenanthrene-d,.-, and
Cg—gamna—BHC.
Ions to identify and measure Clg-Clg-PCBs.
c Ions to identify and measure Clg-Cl^-PCBs and C^-^'^
Ions to identify and measure Clg-Clg-PCBs and chrysene-d^*
e Ions to identify and measure Clg-Cl^Q-PCBs.
-------
-37-
Table 8. Retention Time Data for PCS Isomer Groups and Calibration Congeners
Isomer Group
Monochlorobiphenyls
Dichlorobiphenyls
Trichlorobiphenyls
TetrachlorohLphenyls
Pentachlorobiphenyls
Hexachlorobiphenyls
Heptachlorobiphenyls
Octachlorobiphenyls
Nonachlorobiphenyls
Decachlorobiphenyl •
Approximate
RRT Range*
0.30-0.35
0.38-0.50
0.46-0.64
0.55-0.82
0.64-0.92
0.75-1.1
0.88-1.2
0.99-1.21
1.16-1.28
1.3
Cal . Cong .
Number
1
5
29
50
87
154
188
200
-
209
Cal • Cong
RRTa
0.30
0.43
0.54
0.56
0.80
0.82
0.88
1.03
-
1.3
a Retention time relative to chrysene-d.., with a 30 mX 0.31 mm ID SE-54 fused
silica capillary column and the following GC conditions: splitless injection
at 80°C; hold for 1 min; heat rapidly to 160°C and hold 1 min; increase at
3*C/min to 310«C.
-------
-38-
Table 9. Ions for Selected Ion Monitoring Data Accpiisition for Pesticide Analytes,
Internal Standards and Surrogate Compounds (Ordered by Retention Time)
Ion Analyte/Internal Std/ Approx. Quant.
Set Surrogate Compound (MW) RRT Ion
Alpha-BHC
Beta-BHC
.mma-BHi
Cg-gaanna-BHC (294)
Phenanthr ene-d-j Q (188)
Delta-BHC
Heptachlor
Aldrin
Gannna—BHC
13
Heptachlor epoxide (386)
Gamma-chlordane (406)
Endosulfan I
Alpha-chlorda ne (406)
Trans-nonachlor (440)
Dieldrin
4,4'-DDE
Endrin
Endosulfan II
4,4'-DDD
Endrin aldehyde (378)
Endosulfan sulfate(420)
4,4'-DDT
13C12-4,4'-DDT
Endrin ketone
Chrysene-d-|2
Methoxychlor
(288)
(288)
(288)
(294)
(188)
(288)
(370)
(362)
(386)
(406)
(404)
(406)
(440)
(378)
(316)
(378)
(404)
(318)
(378)
(420)
(352)
(364)
(378)
(240)
(344)
0.43
0.47
0.48
0.48
0.49
0.51
0.5$
0.64
0.70
0.74
0.76
0.76
0.77
0.80
0.81
0.83
0.35
0.87
0.88
0.92
0.93
0.93
0.99
1.00
1.03
219
219
219
225
188
219
272
263
353
373
195
373
409
79
246
81
195
235
67
272
235
247
67
240
227
Ions (Approximate
Relative Abundance)
181 (100), 183 (90), 219 (70)
181 (100), 183 (90), 219 (70)
181 (100), 183 (90), 219 (75)
187 (100), 189 (90) 225 (80), 227 (40)
188 (100), 189 (15)
181 (100), 183 (90), 219 (70)
100 (100), 272 (60), 274 (40)
66 (100), 263 (40), 265 (25)
81 (100), 353 (80), 355 (65)
373 (100), 375 (.95)
195 (100), 339 (50), 341 (35)
373 (100), 375 (95)
409 (100), 407 (85)
79 (100), 263 (10), 108 (15)
246 (100), 248 (65)
81 (100), 263 (75)
195 (100), 339 (50), 341 (35)
235 (100), 237 (65), 165 (65)
67 (100), 345 (30)
272 (100), 274 (80), 387 (50)
235 (100), 237 (65), 165 (65)
247 (100), 249 (65)
67 (100), 317 (50)
240 (100), 241 (20)
227 (100), 228 (15)
-------
-39-
Table 10. Ion Sets for Selected Ion Monitoring of Pesticide Analytes, Internal
Standards and Surrogate Compounds (Ordered by Retention Time)
Ion Set
No, 1
66
100
181
183
187
188
189
219
225
227
263
265
272
274
Monitored
Compounds
Alpha-BHC
Beta-BHC
Delta-BHC
Gamma— BHC
Cg-gamma-BHC
Phenanthrene-d-j Q
Heptachlor
Aldrin
Ion Set
No. 2
79
81
108
195
246
248
263
339
341
353
355
373
375
407
409
Monitored
Compounds
Heptachlor
epoxide
Alpha-chlordane
Gamma-chlordane
Endosulfan I
Trans-nonachlor
Dieldrin
4, 4 '-DDE
Endrin
Endosulfan II
Ion Set Monitored
No. 3 Compounds
67 4 ,4 '-ODD
165 Endrin aldehyde
227 Endosulfan sulfate
228 4,4'-DDT
235 13C12-4,4'-DDT
237 Endrin fcetone
240 Chrysene-di2
241 Methoxychlor
247
249
272
274
317
345
387
14 ions, 8 compounds
15 ions, 9 compounds
15 ions 3 compounds
-------
-40-
Table 11. Known Relative Abundances of Ions in PCS Molecular Ion Clusters*
m/z
Relative
Intensity
Monochlorobiphenyls
188 100
'189 13.5
190 33.4
192 4.41
Dichlorobiphenyls
222 100
223 13.5
224 66.0
225 8.82
226 11.2
227 1.44
Trichlorobiphenyls
256 100
257 13.5
258 98.6
259 13.2
260 32.7
261 . 4.31
262 3.73
263 0.47
Tetrachlorobiphenyls
290 76.2
291 10.3
292 100
293 13.4
294 49.4
295 6.57
296 11.0
297 1.43
298 0.95
Penta chlorobiphenyIs
324 61.0
325 8.26
326 100
327 13.5
328 65.7
329 8.78
330 21.7
331 2.86
332 3.62
333 0.47
334 0.25
m/z
Relative
Intensity
Hexachlorobiphenyls
358 50.9
359 6.89
360 • 100
361 13.5
362 82.0
363 11.0
364 . 36.0
365 4.77
366 8.92
367 1.17
368 1.20
369 0.15
HeptachlorobiphenyIs
392 43.7
393 5.91
394 100
395 13.5
396 98.3
397 ' 13.2
398 53.8
399 7.16
400 17.7
401 2.34
402 3.52
403 0.46
404' 0.40
Octachlorobiphenyls
426 33.4
427 4.51
428 87.3
429 11.8
430 100
431 13.4
432 65.6
433 8.76
434 26.9
435 3.57
436 7.10
437 0.93
438 1.18
439 0.15
440 0.11
m/z
Relative
Intensity
Nonachlorobiphenyls
460 26.0
461 3.51
462 76.4
463 10.3
464 100
465 13.4
466 76.4
467 10.2
468 37.6
469 5.00
470 12.4
471 1.63
472 2.72
473 0.35
474 0.39
Decachlorobipheny1
494 20.8
495 2.81
496 68.0
497 9.17
498 100
499 13.4
500 87.3
501 11.7
502 50.0
503 6.67
504 19.7
505 2.61
506 5.40
507 0.71
508 1.02
509 0.13
Source: Rote and Morris (9)
-------
-41-
Table 12. Quantitation, Confirmation, and Interference Check Ions for PCBs,
Internal Standards, and Surrogate Compounds
Analyte/
Norn. Quant. Confirm. Expected
M-70 Interference
Accept. Confirm. Check Ions
Internal Std.
PCB I some r Group
C11
C12
C13
C14
C15
Cl6
C17
Clg
C19
Clin
. MW
188
222
256
290
324
358
392
426
460
494
Ion
188
222
256
292
326
360
394
430
464
498
Ion
190
224
258
290
324
362
396
428
466
500
Ratioa
3.0
1.5
1.0
1.3
1.6
1.2
1.0
*
1.1
1.3
1.1
Ratio*
2.5-3.5
1.3-1.7
0.8-1.2
1.1-1.5
1.4-1.8
1.0-1.4
0.8-1.2
0.9-1.3
1.1-1.5
0.9-1.3
Ion
152b
152
186
220
254
288
322
356
390
424
M+70
256
292
326
360
394
430
464
498
-
.
M+35
222
256
290
326
360
394
430
464
498
_
Internal standards
Chrysene-d|2 240
Phenanthrene-dfQ 188
Surrogate compounds
13
Cg-gamma-BHC 294
'C-j^^'-DOT 364
240
188
187
247
241
189
»
189
249
5.1
6.6
1.0
1.5
4.3-5.9
6.0-7.2
0.8-1.2
1.3-1.7
a Ratio of quantitation ion-to confirmation ion
^ Monodichlorobiphenyls lose HC1 to produce an ion at m/z 152.
-------
-42-
Table 13. Correction for Interference of PCS Containing Two Additional Chlorines
Candidate
Ion Measured
Quant. Confirm, to Determine
% of Meas. Ion Area to
be Subtracted from
Isomer Group
Trichlorobiphenyls
Tetrachlorobiphenyls "
Pentachlorobiphenyls
Hexachlorobiphenyls
Heptachlorobiphenyls
Ion
256
292
326
360
394
Ion
258
290
324
362
396
Interference
254
288
322
356
390
Quant
Ion Area
99%
65%
108%
161%
225%
Confirm.
Ion Area
33%
131%
164%
71%
123%
Table 14. Correction for Interference of PCS Containing One Additional Chlorine
Candidate
Isomer Group
Dichlorobiphenyls
Trichlorobiphenyls
Te tr achlorobipheny Is
Pentachlorobipheny Is
Hexachlorobiphenyls
Heptachlorobipheny Is
Oct achlorobipheny Is
Quant.
Ion
222
256
292
326
360
394
430
to Determine
Interference
221
255
289
323
357
391
425
to be Subtracted
from Quant. Ion Area
13.5%
13.5%
17.4%
22.0%
26.5%
30.9%
40.0%
-------
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