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EPA
815-
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00-
001

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                                             EPA #815/6-00/001

METHOD 515.4    DETERMINATION OF CHLORINATED ACIDS IN DRINKING
                 WATER BY LIQUID-LIQUID MICROEXTRACTION,
                 DERIVATIZATION, AND FAST GAS CHROMATOGRAPHY WITH
                 ELECTRON CAPTURE DETECTION
                                 Revision 1.0

                                  April 2000


S.C. Wendelken, M.V. Bassett, T.A. Dattilio, and B.V. Pepich (IT Corporation)
D.J. Munch (US EPA, Office of Ground Water and Drinking Water) - Method 515.4, Revision 1.0
(2000)

A. M. Pawlecki-Vonderheide (1C1) and D.J. Munch  (US EPA, Office of Ground Water and Drinking
Water) - Method 515.3, Revision 1 0 (1996)

J.W. Hodgeson - Method 515.2. Revision 1.0 (1992)

R.L. Graves - Method 515.1. Rc\ ision 4.0 (1989)

T. Engels (Batelle Columbus Laboratory) and D.J. Munch  (US EPA, Office of Ground Water and
Drinking Water) - National Pesticide Survey Method 3, Revision 3.0 (1987)

J.W. Hodgeson - Method 515. Rcxision 2.0 (1986)
                         TECHNICAL SUPPORT CENTER
               OFFICE OF GROUND WATER AND DRINKING WATER
                  U. S. ENVIRONMENTAL PROTECTION AGENCY
                           CINCINNATI, OHIO 45268
                                   515.4-1

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METHOD 515.4    DETERMINATION OF CHLORINATED ACIDS IN DRINKING
                 WATER BY LIQUID-LIQUID MICROEXTRACTION,
                 DERIVATIZATION, AND FAST GAS CHROMATOGRAPHY WITH
                 ELECTRON CAPTURE DETECTION

1.     SCOPE AND APPLICATION

      1.1    This is a fast gas chromatography (GC) method for the determination of chlorinated
           acids in drinking waters. Accuracy, precision, and Detection Limit data have been
           generated for the following compounds in reagent water and finished ground and
           surface waters.
Analyte
Acifluorfen(a)
Bentazon
Chloramben
2,4-D
Dalapon
2,4-DB
Dacthal acid metabolites"5'
Dicamba
3,5-Dichlorobenzoic acid
Dichlorprop
Dinoseb
Pentachlorophenol
Picloram
2,4,5-T
2,4,5-TP (Silvex)
Quinclorac
Chemical Abstracts Service
Registry Number
50594-66-6
25057-89-0
133-90-4
94-75-7
75-99-0
94-82-6

1918-00-9
51-36-5
120-36-5
88-85-7
87-86-5
1918-02-1
93-76-5
93-72-1
84087-01-4
u)The herbicide Lactofen will be quantitated as Acifluorfen as their
structures represent different esters of the same acid herbicide.
(b) Dacthal mono-acid and di-acid metabolites are included in method
scope; Dacthal di-acid was used for validation studies.
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       1.2    This method is also applicable to the determination of salts and esters of analyte
             acids.  The form of each acid is not distinguished by this method. Results are
             calculated and reported for each listed analyte as the total free acid. This method is
             able to quantify the mono- and di-acid forms of Dacthal but not the parent compound.
       1.3    Detection Limits are compound, instrument, and matrix dependent. The Detection
             Limit is defined as the statistically calculated minimum amount that can be measured
             with 99% confidence that the reported value is greater than zero."1 Experimentally
             determined Detection Limits for the above listed analytes are provided in Section  17,
             Table 4.  The Detection Limit differs from, and is usually lower than (but never
             above), the minimum reporting level (MRL) (Sect. 3.17).  The concentration range
             for target analytes in this method was evaluated between 0.5 ug/L and 20 ug/L for a
             40-mL sample. Precision and accuracy data are presented in Section 17, Tables 5  -
             10.

       1.4    This method is restricted to use by or under the supervision of analysts skilled in
             liquid-liquid extractions, derivatization procedures and the use of GC and
             interpretation of gas chromatograms. The method was developed using fast GC but
             "conventional" GC may be used as long as the laboratory  meets the requirements of
             the IDC (Sect. 9.2).

2.      SUMMARY OF METHOD

       2.1    A 40-mL volume of sample is adjusted to pH s 12 with 4 N sodium hydroxide and
             allowed to sit for one hour at room temperature to hydrolyze derivatives.  Following
             hydrolysis, a wash step using a hexane:MtBE mixture is performed as a sample
             cleanup and to remove Dacthal. The aqueous sample is then acidified with sulfuric
             acid to a pH of less than 1 and extracted with 4-mL of methyl  tert-butyl ether (MtBE).
             The chlorinated acids that have been partitioned into the MtBE are then converted to
             methyl esters by derivatization  with diazomethane. The target esters are separated
             and identified by fast capillary column gas chromatography using an electron capture
             detector (GC/ECD). Analytes are quantified using a procedural standard calibration
             technique with an internal standard.

             NOTE: since many of the analytes contained in this method are applied as a variety of
             esters and salts, it is imperative to hydrolyze them to the parent acid prior to
             extraction.

3.      DEFINITIONS

       3.1    EXTRACTION BATCH - A set of up to 20 field samples (not including QC

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      samples) extracted together by the same person(s) during a work day using the same
      lots of solvents, surrogate solution, and fortifying solutions. Required QC samples
      include Laboratory Reagent Blank, Laboratory Fortified Matrix, and either a Field
      Duplicate or Laboratory Fortified Matrix Duplicate.

3.2    ANALYSIS BATCH - A set of samples that is analyzed on the same instrument
      during a 24-hour period that begins and ends with the analysis of the appropriate
      Continuing Calibration Check standards (CCC). Additional CCCs may be required
      depending on the length of the analysis batch and/or the number of Field Samples.

3.3    INTERNAL STANDARD (IS) - A pure analyte added to an extract or standard
      solution in a known amount and used to measure the relative responses of other
      method analytes and surrogates. The internal standard must be an analyte that is not a
      sample component.

3.4    SURROGATE ANALYTE (SUR) - A pure analyte, which is extremely unlikely to
      be found in any sample, and which is added to a sample aliquot in a known amount
      before extraction or other processing, and is measured with the same procedures used
      to measure other sample components.  The  purpose of the SUR is to monitor method
      performance with each sample.

3.5    LABORATORY  REAGENT BLANK (LRB) - An aliquot of reagent water or other
      blank matrix that  is treated exactly  as a sample including exposure to all glassware,
      equipment, solvents, reagents, sample preservatives, internal standards, and
      surrogates that arc used in ihe extraction batch. The LRB is used to determine if
      method analytes or other interferences are present in the laboratory environment, the
      reagents, or the apparatus.

3.6    LABORATORY  FORTIFIED BLANK (LFB) - An aliquot of reagent water or other
      blank matrix to \vhich known quantities of the method analytes and all the
      preservation compounds are added. The LFB is analyzed exactly like a sample, and
      its purpose is to determine whether the methodology is in control, and whether the
      laboratory is capable of making accurate and precise measurements.

3.7    LABORATORY  FORTIFIED SAMPLE MATRIX (LFM) - An aliquot of an
      environmental sample to which known quantities of the method analytes and all the
      preservation compounds are added in the laboratory. The LFM is analyzed exactly
      like a sample, and its purpose is to determine whether the sample matrix contributes
      bias to the analytical results.  The background concentrations of the analytes in the
      sample matrix  must be determined  in a separate aliquot and the measured values in
      the LFM corrected for background concentrations.

3.8    LABORATORY FORTIFIED SAMPLE MATRIX DUPLICATE (LFMD) - A

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      second aliquot of the Field Sample used to prepare the LFM which is fortified,
      extracted and analyzed identically. The LFMD is used instead of the Field Duplicate
      to access method precision and accuracy when the occurrence of target analytes is
      low.

3.9   LABORATORY DUPLICATES (LD1 and LD2) - Two aliquots of the same sample
      taken in the laboratory and analyzed separately with identical procedures. Analyses
      of LD1 and LD2 indicate precision associated with laboratory procedures, but not
      with sample collection, preservation, and storage procedures.

3.10  FIELD DUPLICATES (FD1 and FD2)  - Two separate samples collected at the same
      time and place under identical circumstances, and treated exactly the same throughout
      field and laboratory procedures. Analyses of FD1 and FD2 give a measure of the
      precision associated with sample collection, preservation, and storage, as well as with
      laboratory procedures.

3.11   STOCK STANDARD SOLUTION (SSS) - A concentrated solution containing one
      or more method analytes prepared in the laboratory using assayed reference materials
      or purchased from a reputable commercial source.

3.12  PRIMARY DILUTION STANDARD SOLUTION (PDS) - A solution containing
      method analytes prepared in the laboratory from stock standard solutions and diluted
      as needed to prepare calibration solutions and other needed analyte solutions.

3.13  CALIBRATION STANDARD (CAL) - A solution prepared from the primary
      dilution standard solution or stock standard solutions and the internal standards and
      surrogate analytes. The CAL solutions are used to calibrate the instrument response
      with respect to analyte concentration.

3.14  CONTINUING CALIBRATION CHECK (CCC) - A calibration standard containing
      one or more method analytes, which  is analyzed periodically to verify the accuracy of
      the existing calibration for those analytes.

3.15  QUALITY CONTROL SAMPLE (QCS) - A solution of method analytes of known
      concentrations that is  obtained from a source external to the laboratory and different
      from the source of calibration standards. It is used to check standard integrity.

3.16  DETECTION LIMIT - The minimum concentration of an analyte that can be
      identified, measured and reported with 99% confidence that the analyte concentration
      is greater than zero (Section 9.2.4).  This is a statistical determination of precision.
      Accurate quantitation is not expected at this level(l).

3.17  MINIMUM REPORTING LEVEL (MRL) - The minimum concentration that can be

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             reported as a quantified value for a target analytc in a sample following analysis. This
             defined concentration can be no lower than the concentration of the lowest continuing
             calibration standard for that analyte, and can only be used if acceptable quality control
             criteria for this standard are met.

       3.18   PROCEDURAL STANDARD CALIBRATION - A calibration method where
             aqueous calibration standards are prepared and processed (e.g., purged, extracted,
             and/or denvatized) in exactly the same manner as a sample. All steps in the process
             from addition of sampling preservatives through instrumental analyses are included in
             the calibration. Using procedural standard calibration compensates for any
             inefficiencies in the processing procedure.

       3.19   MATERIAL SAFETY DATA SHEET (MSDS) - Written information provided by
             vendors concerning a chemical's toxicity, health hazards, physical properties, fire,
             and reactivity data including storage, spill, and handling precautions.

4.     INTERFERENCES

       4.1    All glassware must be meticulously cleaned. Wash glassware with detergent and tap
             water, rinse with tap water,  followed by reagent water. A final rinse with  solvents
             may be needed.  In place of a solvent rinse, non-volumetric glassware can be muffled
             at 400 C for 2 hours. Volumetric glassware should not be heated in an oven above
             120 C. Store inverted or capped with aluminum foil.

       4.2    Method interferences may be caused by contaminants in solvents, reagents (including
             reagent water), sample bottles and caps, and other sample processing hardware that
             lead to discrete artifacts and/or elevated  baselines in the chromatograms.  All items
             such as these must be routinely demonstrated to be free from interferences (less than
             V3 the MRL for each target) under the conditions of the analysis by analyzing
             laboratory  reagent blanks as described in Section 9. Subtracting blank values from
             sample results is not permitted.

             4.2.1  During method development it was found that sodium  sulfate from several
                     sources contained multiple interferences. After screening several  brands, it
                     was found that a grade suitable for pesticide residue analysis provided the best
                     results.  If the suitability of the available sodium sulfate is in question,  extract
                     and analyze a laboratory reagent blank (section 3.5) to test for interferences
                     prior to sample processing.

       4.3    Matrix interferences may be caused by contaminants that are extracted from the
             sample. The extent of matrix interferences will vary considerably from source to
             source, depending upon the water sampled.
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              4.3.1  During method development a contaminant was found in local ground water
                    and surface water that interfered with the analysis of Dalapon on the primary
                    column.  Due to this mterferent, it was necessary to carefully optimize the
                    chromatographic conditions (see Table 1) A slow initial temperature program
                    was needed to ensure separation of the mterferent from Dalapon  Analyte
                    identifications should be confirmed using the confirmation column specified
                    in Table 2 or another column that is dissimilar to the primary column or by
                    GC/MS if the concentrations are sufficient.

       4.4     Interferences by phthalate esters can pose a major problem in pesticide analysis when
              using an electron capture detector (ECD). These  compounds generally appear in the
              chromatogram as large peaks. Common flexible plastics contain varying amounts of
              phthalates that are easily extracted or leached during laboratory operations. Cross
              contamination of clean glassware routinely occurs when plastics are handled during
              extraction steps, especially when solvent-wetted  surfaces are handled. Interferences
              from phthalates can best be minimized by avoiding the use of plastics in the
              laboratory. Exhaustive purification of reagents and glassware may be required to
              eliminate background phthalate contamination.'2 3>
5.     SAFETY
       5.1     The toxicity or carcinogemcity of each reagent used in this method has not been
              precisely defined.  Each chemical compound should be treated as a potential health
              hazard, and exposure to these chemicals should be minimized. 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
              MSDSs should also be made available to all personnel involved  in the chemical
              analysis. Additional references to laboratory safety are available.'4'"

       5.2     Pure standard materials and stock standards of these compounds should be handled
              with suitable protection to skin and eyes, and care should be taken not to breathe the
              vapors or ingest the materials.

       5.3     The toxicity of the extraction solvent, MTBE, has not been well  defined.  Susceptible
              individuals may experience adverse affects upon skin contact or inhalation of vapors.
              Therefore protective clothing and gloves should be used and-MTBE should be used
              only in a chemical fume hood or glove box. The same precaution applies to pure
              standard materials.

       5.4     Diazomethane is a toxic carcinogen which can explode under certain conditions. The
              following precautions must be followed.

              5.4.1   Use the diazomethane generator behind a safety shield in a well ventilated

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                   fume hood. Under no circumstances can the generator be heated above 90C,
                   and all grinding surfaces such as ground glass joints, sleeve bearings and glass
                   stirrers must be avoided. To minimize safety hazards, the diazomethane
                   generator apparatus used in the estenfication procedure (Sect 11.2) produces
                   micro molar amounts of diazomethane in solution.  If the procedure is
                   followed carefully, no possibility for explosion exists.

      5.5    Although hydrogen can be safely used as a carrier gas, the potential for fire or
             explosion does exist if the gas system is mishandled.  If you are unsure of the safety
             guidelines for using hydrogen as a carrier gas, seek advice from your instrument
             manufacturer regarding its use.

6.     APPARATUS AND EQUIPMENT

      6.1    SAMPLE CONTAINERS - Amber glass bottles, approximately 40 mL, fitted with
             PTFE (polytetrafluoroethylene) lined screw caps.

      6.2    EXTRACTION VIALS - 60-mL clear glass vials with PTFE lined screw caps.

      6.3    AUTOSAMPLER VIALS -  2.0-mL vials with screw or crimp cap and a PTFE faced
             seal.

      6.4    STANDARD SOLUTION STORAGE CONTAINERS - 10 to 20-mL amber glass
             vials with PTFE lined screw caps.

      6.5    CLEAR VIALS - 7-mL glass, disposable, with PTFE lined screw caps for extract
             drying and derivatization.

      6.6    PASTEUR PIPETTES - Glass, disposable.

      6.7    PIPETTES - Class A, 2.0-mL and 4.0-mL glass, or adjustable volume dispensers.

      6.8    VOLUMETRIC FLASKS - Class A, suggested sizes include 5 mL. 10 mL, 100 mL.

      6.9    MICRO SYRINGES - Various sizes.

      6.10   BALANCE - Analytical, capable of weighing to the nearest 0.0001 g.

      6.11   DIAZOMETHANE GENERATOR - See Figure 4 for a diagram of an all glass
             system custom made for these validation studies. Micro molar generators are also
             available from commercial sources (Aldrich Cat.#:  Z10.889-8 or equivalent).

      6.12   GAS CHROMATOGRAPH - Capillary GC (Hewlett Packard Model 6890 or

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             equivalent), if the fast GC option is used the modifications should include a high
             pressure (> 50 psi) split/splitless injector, fast temperature ramp oven (50 C/mmute)
             and a low volume (150 nL ) micro ECD detector  Additionally, a data system capable
             of fast sampling (20 points/peak) is required.

      6.13   PRIMARY GC COLUMN-RTX-1701, 180 urn i.d., fused silica capillary with
             chemically bonded (14% cyanopropylphenyl-methylpolysiloxane), or equivalent
             bonded, fused silica column.

      6.14   CONFIRMATION GC COLUMN - DB-5, 180 urn i.d., fused silica capillary with
             chemically bonded (5% phenyl-methylpolysiloxane), or equivalent bonded, fused
             silica column.

7.     REAGENTS AND STANDARDS

      7.1    REAGENTS AND SOLVENTS - Reagent grade or better chemicals should be used
             in all analyses. 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 determined that the reagent is of sufficiently high
             punty to permit its use without lessening the quality of the determination.

             7.1.1   REAGENT WATER - Purified water which does not contain any measurable
                   quantities of any target analytes or interfering compounds greater than  1/3 the
                   MRL for each compound of interest.

             7.1.2   METHYL tert-BUTYL ETHER (MtBE) - High purity, demonstrated to be
                   free from analytes and interferences (HPLC grade or better).

             7.1.3  ACETONE - High purity, demonstrated to be free from analytes and
                   interferences (HPLC grade or better).

             7.1.4  CARBITOL (DIETHYLENE GLYCOL MONOETHYL ETHER) - High
                   purity, demonstrated to be free from analytes and interferences (HPLC grade
                   or better).

             7.1.5  HEXANE:MtBE (90:10, v/v) WASH SOLVENT - High purity, unpreserved,
                   demonstrated to be free from analytes and  interferences (HPLC grade or
                   better).

             7.1.6  HEXANE - High punty, demonstrated to be free from analytes and
                   interferences (HPLC grade or better).
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      7.1.7   SODIUM SULFATE, Na2SO4 - Pesticide grade, granular, anhydrous.
            Interferences have been observed when lower quality grades have been used.
            If interferences are observed, it may be necessary to heat the sodium sulfate in
            a shallow tray at 400 C for up to 4 hr to remove phthalates and other
            interfering organic substances. Alternatively, it can be extracted with
            methylene chloride in a Soxhlet apparatus for 48 hr. Store in a capped glass
            bottle rather than a plastic container.

      7.1.8  ACIDIFIED SODIUM SULFATE - Acidi fy by slurrying 100 g of muffled
            sodium sulfate with enough ethyl ether to just cover the solid. Add 0.5-mL
            concentrated sulfunc acid dropwise while mixing thoroughly. Remove the
            ether under vacuum. Mix 1 g of the resulting solid with 5-mL of reagent
            water and measure the pH of the mixture The pH must be below pH 4. Store
            in a desiccator or  at 100C to keep the reagent dry.

      7.1.9  COPPER II SULFATE PENTAHYDRATE, CUS04-5H,0 - ACS reagent
            grade or better.

      7.1.10 4 N SODIUM HYDROXIDE SOLUTION - Dissolve 16 g sodium hydroxide
            (NaOH) pellets (ACS grade or equivalent) in reagent water and dilute to 100
            mL.

      7.1.11 POTASSIUM HYDROXIDE SOLUTION (37%, w/v) - Dissolve 37 g of
            potassium hydroxide (KOH) pellets (ACS grade or equivalent) in reagent
            water and dilute to 100 mL.

      7.1.12 SODIUM SULFITE, Na,S03 - ACS reagent grade, used as a dechlonnating
            agent in this method.

      7.1.13 DIAZALD SOLUTION - Prepare a solution containing 5 g diazald (ACS
            reagent grade) in 50 mL of a 50:50 (v/v) mixture of MtBE and carbitol. This
            solution is stable for one  month or longer when stored at 4 C in an amber
            bottle with a PTFE lined  screw cap.

      7.1.14 SULFURIC ACID -  Concentrated, ACS reagent grade.

      7.1.15 SILICA GEL - ACS reagent grade, 35-60 mesh.

      7.1.16 HYDROGEN - 99.999% pure or better, GC carrier gas.

      7.1.17 NITROGEN - 99.999% pure or better, ECD make-up gas

7.2    STANDARD SOLUTIONS - When a compound purity is assayed to be 96% or

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greater, the weight can be used without correction to calculate the concentration of the
stock standard. Solution concentrations listed in this section were used to develop
this method and are included as an example.  Standards for sample fortification
generally should be prepared in the smallest volume that can be accurately measured
to minimize the addition of organic solvent to aqueous samples.  Laboratories
should use standard QC procedures to determine when Standard Solutions
described in this section need to be replaced.

7.2.1  INTERNAL STANDARD SOLUTIONS - 4,4'-Dibromooctafluorobiphenyl,
      99+% is used as an internal standard for the method.  This compound has been
      shown to be an effective internal standard for the method  analytes, but other
      compounds may be used if the QC requirements in Section 9 are met.

      7.2.1.1 INTERNAL STANDARD STOCK SOLUTION (2.0 mg/mL) -
             Prepare an internal standard a stock solution by accurately weighing
             approximately 0.0200 g of neat 4,4'- dibromooctafluorobiphenyl
             material. Dissolve the neat material in MtBE and dilute to volume in a
             10-mL volumetric flask. Transfer the solution to an amber glass vial
             with a PTFE-lined screw cap and store at 5 0 C. The resulting
             concentration of the stock internal standard solution will be
             approximately 2.0 mg/mL.

      7.2.1.2 INTERNAL STANDARD PRIMARY DILUTION STANDARD (2.5
             ug/mL) - Prepare a internal standard fortification  solution at
             approximately 2.5 ug/mL by the addition of 12.5 uL of the stock
             standard to 10 mL of MtBE. Transfer the primary  dilution to an amber
             glass vial with a PTFE lined screw cap and store at <  0 C. The
             solution should  be replaced when ongoing QC indicates a problem.

      7.2.1.3 MtBE EXTRACTION SOLVENT WITH INTERNAL STANDARD
             (2.5 ug/mL) - The internal standard 4,4'-dibromooctafluorobiphenyl is
             added to the extraction solvent prior to analyte extraction to
             compensate for  any volumetric differences encountered during sample
             processing. This solution should be made fresh prior to extraction.
             The addition of 1 mL of the primary dilution standard (2.5 ug/mL) to
             99-mL MtBE results in a final internal standard concentration of 25
             ng/mL. This solution is used to extract the samples (Sect. 11.1).

7.2.2  SURROGATE (SUR) ANALYTE STANDARD SOLUTION -
      2,4-Dichlorophenylacetic acid (99+%) is used as a surrogate compound in this
      method to evaluate the  extraction and denvatization procedures. This
      compound has been shown to be an effective surrogate for the method
      analytes, but other compounds may be used if the QC requirements in Section

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      9 are met.

      7.2.2.1 SURROGATE STOCK SOLUTION (1.0 mg/mL) - Prepare a
             surrogate stock standard solution of 2,4-dichlorophenylacetic acid by
             weighing 0.0100 g of neat material. Dissolve the neat material in
             acetone and dilute to volume in a 10-mL volumetric flask Transfer the
             solution to an amber glass vial with a PTFE lined screw cap and store
             at s 0 C. The resulting concentration of the stock surrogate solution
             will be  1.0 mg/mL.

      7.2.2.2 SURROGATE PRIMARY DILUTION STANDARD/SUR SAMPLE
             FORTIFICATION SOLUTION (100 ug/mL) - Prepare a primary
             dilution standard at approximately 100 ug/mL by the addition of 1 mL
             of the stock standard to 10 mL of acetone.  Transfer the primary
             dilution to an amber glass vial with a PTFE lined screw cap and store
             at s 0 C. Addition of 10 ul, of the primary dilution standard to the
             40-mL aqueous sample results in a surrogate concentration of 25
             ng/mL. The solution should be replaced when ongoing QC indicates a
             problem.

7.2.3  ANALYTE STANDARD SOLUTIONS - Obtain the analytes listed in the
      table in Section 1.1 as neat or solid free acid standards or as commercially
      prepared ampulized solutions from a reputable standard manufacturer.  The
      use of pre-methylated standards is not allowed for the preparation of analyte
      standards. Prepare the Analyte Stock and Primary Dilutions Standards as
      described below.

      7.2.3.1 ANALYTE STOCK STANDARD SOLUTION - Prepare separate
             stock standard solutions for each analyte of interest at a concentration
             of 1 -5 mg/mL in acetone. Method analytes may be obtained as neat
             materials or ampulized solutions (> 99% purity) from a number of
             commercial suppliers.  These stock standard solutions should be stored
             at < 0 C.

             7.2.3.1.1   For analytes which are solids in their pure form, prepare
                       stock standard solutions by accurately weighing
                       approximately 0.01 to 0.05 grams of pure material in a 10-
                       mL volumetric flask. Dilute to volume with acetone.

             7.2.3.1.2   For analytes which are liquid in their pure form at room
                       temperature, place about 9.8 mL of acetone into a 10-mL
                       volumetric flask. Allow the flask to stand, unstoppered, for
                       about 10 minutes to allow solvent film to evaporate from

                           515.4-12

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                 the inner walls of the volumetric, and weigh to the nearest
                 0.1 mg.  Use a 10-uL syringe and immediately add 10.0 uL
                 of standard material to the flask by keeping the syringe
                 needle just above the surface of the acetone. Be sure that
                 the standard material falls dropwise directly into the
                 acetone without contacting the inner wall of the volumetric.
                 Calculate the concentration in milligrams per milliliter
                 from the net gain in weight.  Dilute to volume, stopper,
                 then mix by inverting the flask several times.

7.2.3.2 PRIMARY DILUTION STANDARD (PDS) - Prepare the primary
       dilution standard solution by combining and diluting stock standard
       solutions with acetone. This primary dilution standard solution should
       be stored at s 0 C. As a guideline to the analyst, the analyte
       concentrations used in the primary dilution standard solution during
       method development are given below.
Analyte
Acifluorfen
Bentazon
Chloramben
2,4-D
Dalapon
2,4-DB
Dacthal acid metabolites
Dicamba
3,5-Dichlorobenzoic acid
Dichlorprop
Dinoseb
Pentachlorophenol
Picloram
2,4,5-T
2,4,5-TP (Silvex)
Quinclorac
Concentration
(ug/mL)
5.0
10
5.0
10
10
10
5.0
5.0
5.0
10
10
1.0
5.0
2.5
2.5
5.0
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                          This primary dilution standard is used to fortify reagent water for
                          calibration standards. The lowest calibration standard concentration
                          must be at or below the MRL of each analyte. The concentrations of
                          the other standards should define a range containing the expected
                          sample concentrations or the working range of the detector.

             7.2.4   CALIBRATION STANDARDS (CAL) - A five-point calibration curve is to
                    be prepared by fortifying reagent water with the primary dilution standard. A
                    designated amount of each calibration standard in acetone is spiked into
                    separate 40-mL aliquots of reagent water to produce a calibration curve
                    ranging from below or at the MRL to approximately 10-20 times the lowest
                    calibration level. These aqueous calibration standards should be treated like
                    samples and therefore require the addition of all preservation and other
                    reagents. They are extracted by the procedure described in Section 11. The
                    calibration standard solutions in acetone should be stored at < 0 C.

8.      SAMPLE COLLECTION. PRESERVATION. AND STORAGE

       8.1    SAMPLE BOTTLE PREPARATION

             8.1.1   Grab samples must be collected in accordance with conventional sampling
                    practices'7' using amber glass containers with  PTFE lined screw caps and
                    capacities of at least 40 mL.

             8.1.2   Add sodium sulfite crystals (approximately 2 mg/40 mL of sample) to the
                    sample bottle prior to collecting the sample. This reagent eliminates residual
                    chlorine in the sample.

       8.2.   SAMPLE COLLECTION

              8.2.1  Fill sample bottles but take care not to flush out the sodium sulfite.  Because
                    the target analytes of this method are not volatile, it is not necessary to ensure
                    that the sample bottles are completely headspace free.

              8.2.2  When sampling from a water tap, remove the aerator so that no air bubbles
                    will be trapped in the sample. Open the tap and allow the system to flush until
                    the water temperature has stabilized (usually about 3-5 minutes).  Collect
                    samples from the flowing system.

             8.2.3   When sampling from an open body of water, fill a 1 quart wide-mouth bottle
                    or 1 liter beaker with sample from a representative area,  and carefully fill
                    sample vials from the container.

                                         515.4-14

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             8.2.4   After collecting the sample, seal the bottle and agitate by hand for 15 seconds.

       8.3    SAMPLE SHIPMENT AND STORAGE - All samples should be iced during
             shipment and must not exceed 10 C during the first 48 hours after collection.
             Samples should be confirmed to be at or below 10 C when they arc received at the
             laboratory.  Samples stored in the lab must be held at or below 6 "C and protected
             from light until extraction. Samples should not be frozen.

       8.4    SAMPLE AND EXTRACT HOLDING TIMES - Because of the several pH
             adjustments made to the samples in the course of this method, the addition of organic
             or inorganic biocides, including hydrochloric acid, have been omitted. The analyst
             should be aware of the potential for the biological degradation of the analytes.
             Accordingly, samples should be extracted as soon as possible.  Samples must be
             extracted within 14 days. Extracts must be stored at   0C or less and protected from
             light in glass vials with PTFE lined caps.  Holding time studies indicate that the
             analytes are stable for up to 21 days in the extracts.

9.      QUALITY CONTROL

       9.1    Quality control (QC)  requirements include the Initial Demonstration of Capability
             (Sect. 17, Table 13). the determination of the Delection Limit, and subsequent
             analysis in each analysis batch of a Laboratory Reagent Blank (LRB), Continuing
             Calibration Check Standards (CCC), a Laboratory Fortified Sample Matrix (LFM),
             and either a Laboraiory Fortified Sample Matrix Duplicate (LFMD) or a Field
             Duplicate Sample. This section details the specific requirements for each QC
             parameter.  The QC criteria discussed in the following sections are summarized in
             Section 17. Tables 13 and 14. These criteria are considered the minimum acceptable
             QC criteria, and laboratories are encouraged to institute additional QC practices to
             meet their specific needs.

             9.1.1   Process all quality control samples through all steps of Section 11, including
                    hydrolysis and methylation. Sample preservatives as described in Section 8.1
                    must be added prior to extracting and analyzing the quality control samples.

       9.2    INITIAL DEMONSTRATION OF CAPABILITY (IDC) - Requirements for the
             Initial Demonstration of Capability are described  in the following sections and
             summarized in Section 17, Table 13.

             9.2.1   INITIAL DEMONSTRATION OF LOW  SYSTEM BACKGROUND -
                    Before any samples are analyzed, it must be demonstrated that a laboratory
                    reagent blank (LRB) is reasonably free of contamination and that the criteria
                    in Section 9.4 are met. .Process all quality control samples through all all steps
                    of the method (Sect. 9.1.1).

                                         515.4-15

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9.2.2   INITIAL DEMONSTRATION OF PRECISION - Prepare, extract, and
       analyze  4-7 replicate LFBs fortified at 5 ug/L, or near the mid-range of the
       initial calibration curve. Process all quality control samples through all all
       steps of the method (Sect. 9.1.1). The relative standard deviation (RSD) of the
       results of the replicate analyses must be less than 20%.

9.2.3   INITIAL DEMONSTRATION OF ACCURACY - Using the same set of
       replicate data generated for Section 9.2.2. calculate average recovery.  The
       average recovery of the replicate values must be within  20% of the true
       value.

9.2.4   DETECTION LIMIT - Prepare, extract and analyze at least seven replicate
       LFBs at a concentration estimated to be near the Detection Limit, over a
       period of at least three days (both extraction and analysis should be conducted
       over at least three days) using the procedure described in Section 11.  Process
       all quality control samples through all all steps of the  method (Sect. 9.1.1).
       The fortification level may be estimated by selecting a concentration with a
       signal of 2 to 5 times the noise level but must  be at or below the laboratory's
       MRL (Sect. 9.3). The appropriate concentration will be dependent upon the
       sensitivity of the GC/ECD system being used.  Sample preservatives as
       described in Section 8.1 must be added to these samples. Calculate the
       Detection Limit using the equation

             Detection Limit = St(n.,  , .aipha=ow)

             where
             l(n-1 i -alpha = 099)= Students t value for the 99% confidence level with n-
              1 degrees of freedom,
             n = number of replicates, and
             S = standard deviation of replicate analyses.

       NOTE: Calculated Detection Limits need only be less than 1/3 of the
       laboratory's MRL to be considered acceptable. Do not subtract blank values
       when performing Detection Limit calculations.  The Detection Limit is a
       statistical determination of precision only.'0 If the Detection Limit replicates
       are fortified at a low enough concentration, it  is likely that they will  not meet
       precision and accuracy criteria, and may result in a calculated Detection Limit
       that is higher than the fortified concentration

9.2.5   METHOD MODIFICATIONS - The analyst  is permitted to modify GC
       columns, GC conditions, internal standards or surrogate standards, but each
       time such method modifications are made, the analyst must repeat the
       procedures of the IDC (Sect. 9.2).

                            515.4-16

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9.3    Minimum Reporting Level (MRL) - The MRL is the threshold concentration of an
       analyte that a laboratory can expect to accurately quantitate in an unknown sample.
       The MRL should not be established at an analyte concentration that is less than either
       three times the Detection Limit or a concentration which would yield a response less
       than a signal-to-noise (S/N) ratio of five. Depending upon the study's data quality
       objectives it may be set at a higher concentration.  Although the lowest calibration
       standard must be at or below the MRL, the MRL must never be established at a
       concentration lower than the lowest calibration standard.

9.4    LABORATORY REAGENT BLANK (LRB) - An LRB is required with each
       extraction batch (Sect. 3.1) of samples to determine the background system
       contamination. If the LRB produces a peak within the retention time window of any
       analyte that would prevent the determination of that  analyte, determine the source of
       contamination and eliminate the interference before processing samples. Background
       contamination must be reduced to an acceptable level before proceeding.  Background
       from method analytes or contaminants that interfere  with the measurement of method
       analytes must be below 1/3 of the MRL. If the target analytes are detected in the LRB
       at concentrations equal to or greater than this level,  then all data for the problem
       analyte(s) must be considered invalid for all samples in the extraction batch.

9.5    CONTINUING CALIBRATION CHECK (CCC) - A standard prepared in the same
       extraction batch as the samples of interest that contains all compounds of interest and
       is extracted in the same manner as the procedural standards used to prepare the initial
       calibration curve. Calibration checks, prepared with the samples being analyzed, are
       required at the beginning of each day that samples are analyzed, after every ten
       samples, and at the end of any group of sample analyses. See Section 10.3 for
       concentration requirements, frequency requirements, and acceptance criteria.

9.6    LABORATORY FORTIFIED BLANK (LFB) - Since this method utilizes procedural
       calibration standards, which are fortified reagent waters, there is no difference
       between the LFB and the continuing calibration check standard. Consequently, the
       analysis of an LFB is not required ; however the acronym LFB  is used for clarity in
       the IDC.

9.7    INTERNAL STANDARDS (IS) - The analyst must monitor the peak area of each
       internal standard in all injections during each analysis day. The IS  response (as
       indicated by peak area) for any chromatographic run must not deviate by more than 
       50% from the average area measured during the initial calibration for that IS. A poor
       injection could cause the IS area to exceed  these criteria. Inject a second aliquot of
       the suspect extract to determine whether the failure is due to poor injection or
       instrument response drift.
                                  515.4-17

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       9.7.1   If the reinjected aliquot produces an acceptable internal standard response,
              report results for that aliquot.

       9.7.2   If the internal standard area for the reinjected extract deviates greater than
              50% from the initial calibration average, the analyst should check  the
              continuing calibration check standards that ran before and after the sample.  If
              the continuing calibration check fails the criteria of Section 10.3, recalibration
              is in order per Section 10. If the calibration standard is acceptable, extraction
              of the sample should be repeated provided the sample is still within holding
              time.  Otherwise, report results obtained from the reinjected extract, but
              annotate as suspect.

9.8    SURROGATE RECOVERY - The surrogate standard is fortified into the aqueous
       portion of all samples,  LRBs, and LFMs and LFMDs prior to extraction.  It is also
       added to the calibration curve and calibration check standards. The surrogate is a
       means of assessing method performance from extraction to final chromatographic
       measurement

       9.8.1   When surrogate recovery from a sample, blank, or CCC is less than 70% or
              greater than 130%, check (1) calculations to locate possible errors, (2)
              standard solutions for degradation, (3) contamination, and (4) instrument
              performance If those steps do not reveal the cause of the problem, reanalyze
              the extract

       9.8.2   If the extract rcanalysis meets the surrogate recovery criterion, report only
              data for the reanalyzed extract.

       9.8.3   If the extract rcanalysis fails the 70-130% recovery criterion, the analyst
              should check the calibration by injecting the last calibration standard that
              passed I f the calibration standard fails the criteria of Section 98.1,
              recalibration is in order per Section 10.2  If the calibration standard is
              acceptable, extraction of the sample should be repeated provided the sample is
              still within the  holding time. If the sample re-extract also fails the recovery
              criterion, report all data for that sample as suspect/surrogate recovery to
              inform the data user that the results are suspect due to surrogate recovery.

9.9    LABORATORY FORTIFIED SAMPLE MATRIX AND DUPLICATE (LFM AND
       LFMD) - Analyses of LFMs (Sect.  3.7) are required in each extraction batch and are
       used to determine that  the sample matrix  does not adversely affect method accuracy.
       If the occurrence of target analytes in the samples is infrequent, or if historical trends
       are unavailable, a second LFM or LMFD must be prepared, extracted, and analyzed
       from a duplicate field sample used to prepare the LFM to assess method precision.
       Extraction batches that contain LFMDs will not require the analysis of a Field

                                   515.4-18

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Duplicate (Sect. 9.10). If a variety of different sample matrices arc analyzed
regularly, for example, drinking water from groundwater and surface water sources,
method performance should be established for each. Over time, LFM data should be
documented for all routine sample sources for the laboratory.

9.9.1   Within each extraction batch, a minimum of one field sample is fortified as an
       LFM for every 20 samples extracted.  The LFM is prepared by spiking a
       sample with an appropriate amount of Analyte PDS (Sect 7.2.3.2). Select a
       spiking concentration that is at least twice the matrix background
       concentration, if known.  Use historical data and rotate through the designated
       concentrations when selecting a fortifying concentration.

9.9.2   Calculate the percent recovery (R) for each analyte using the equation
             where
             A = measured concentration in the fortified sample,
             B = measured concentration in the unfortified sample, and
             C = fortification concentration.

9.9.3   Analyte recoveries may exhibit matrix bias.  For samples fortified at or above
       their native concentration, recoveries should range between 70 and 130%,
       except for low-level fortification near or at the MRL where 50 to 150%
       recoveries are acceptable.  If the accuracy of any analyte falls outside the
       designated range, and the laboratory performance  for that analyte is shown to
       be in control in the LFB, the recovery is judged to be matrix biased. The
       result for that analyte in the unfortified sample is labeled suspect/matrix to
       inform the data user that the results are suspect due to matrix effects.

9.9.4   If an LFMD is analyzed instead of a Field Duplicate (Sect 9. 1 0), calculate the
       relative percent difference (RPD) for duplicate LFMs (LFM and LFMD) using
       the equation
                  RPD-        - LFMD
                         (LFM+ LFMD)/2
       RPDs for duplicate LFMs should fall in the range of  30% for samples
       fortified at or above their native concentration. Greater variability may be
       observed when LFMs are spiked near the MRL.  At the MRL, RPDs should
       fall in the range of  50% for samples fortified at or above their native

                            515.4-19

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             concentration. If the accuracy of any analyte falls outside the designated
             range, and the laboratory performance for that analyte is shown to be in
             control in the LFB, the recovery is judged to be matrix biased. The result for
             that analyte in the unfortified sample is labeled suspect/matrix to inform the
             data user that the results are suspect due 10 matrix effects

9.10   FIELD DUPLICATES (FD1 AND FD2) - Within each extraction batch, a minimum
       of one Field Duplicate (FD) or LFMD (Sect. 9.9) must be analyzed  FDs check the
       precision associated with sample collection, preservation, storage, and laboratory
       procedures. If target analytes are not routinely observed in field samples, a LFMD
       (Sect. 9.10) should be analyzed to substitute for this requirement. Extraction batches
       that contain LFMDs will not require the analysis of a Field Duplicate.

       9. 1 0. 1 Calculate the relative percent difference (RPD) for duplicate measurements
             (FD1 and FD2) using the equation
                                                     tl00
       9.10.2 RPDs for duplicates should be in the range of  30%.  Greater variability may
             be observed when analyte concentrations are near the MRL.  At the MRL,
             RPDs should fall in the range of  50%  If the accuracy of any analyte falls
             outside the designated range, and the laboratory' performance for that analyte
             is shown to be in control in the LFB, the recovery is judged to be matrix
             biased.  The result for that analyte in the unfortified sample is labeled
             suspect/matrix to inform the data user that the results are suspect due to matrix
             effects.

9.11   QUALITY CONTROL SAMPLES (QCS) - A QCS sample should be analyzed each
       time that new Primary Dilution Standards (Sect 7.2.3.2) are prepared  The source of
       the QCS sample should ideally be a second vendor If a second vendor is not
       available then a different lot of the standard should be used.  Although the use of pre-
       methylated standards is prohibited for prepanng analyte standard solutions, pre-
       methylated standards may be used to prepare the QCS.  The QCS may be injected as
       a calibration standard or fortified into reagent water and analyzed as a LFB. If the
       QCS is analyzed as a continuing calibration, then the acceptance criteria are the same
       as for the CCC.  If the QCS is analyzed as a LFB. then the acceptance criteria are the
       same as for an LFB. If measured analyte concentrations are not of acceptable
       accuracy, check the entire analytical procedure to locate and correct the problem
       source.
                                   515.4-20

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10.     CALIBRATION AND STANDARDIZATION

       10.1   After initial calibration is successful, a Continuing Calibration Check (CCC) is
             required at the beginning and end of each analysis batch, and after every tenth sample
             (Sect. 10.3). Because this is a procedural standard method, the analyst will need to
             make a decision to include an appropriate number of CCCs or an entire initial
             calibration curve with each extraction batch.  Initial calibration should be repeated
             each time a major instrument modification or maintenance is performed.

       10.2   INITIAL CALIBRATION

             1 0.2. 1 Establish GC operating parameters equivalent to the suggested specifications
                    in Section  17, Table 1 . The GC system must be calibrated using the internal
                    standard (IS) technique. Other columns or conditions may be used if
                    equivalent or better performance can be demonstrated.

             10.2.2 Prepare a set of at least 5 calibration standards as described in Section 7.2.4.
                    The lowest concentration of calibration standard must be at or below the
                    MRL, which will depend on system sensitivity.

             10.2.3 CALIBRATION - Use the GC data system software to generate a linear
                    regression or quadratic calibration curve using the internal standard. The
                    analyst may choose whether or not to force zero to obtain a curve that best fits
                    the data. Examples of common GC system calibration curve options are:
                    1) Ax /A. vs Q, /Q1S and 2) RRF vs Ax /A,,

                    where:
                           Ax  =      integrated peak area of the analyte,
                           A15 =       integrated peak area of the internal standard,
                           Qx  =      quantity of analyte injected  in ng or concentration units,
                                     and
                           QIS =       quantity of internal standard injected in ng or concentration
                                     units.

             10.2.4 As an alternative, concentrations may be calculated through the use of average
                    relative response factor (RRF).  Calculate the RRFs using the equation
                               RRF


                    Average RRF calibrations may only be used if the RRF values over the


                                         515.4-21

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             calibration range are relatively constant (<30% RSD). Average RRF is
             determined by calculating the mean RRF of a minimum of five calibration
             concentrations.

       10.2.5 Acceptance criteria for the calibration of each analyte is determined by
             calculating the concentration of each analyte and surrogate in each of the
             analyses used to generate the calibration curve or average RRF. Each
             calibration point, except the lowest point, for each analyte must calculate to be
             70-130% of its true value. The lowest point must calculate to be 50-150% of
             its true value. If this criteria cannot be met, reanalyze the calibration
             standards, restrict the range  of calibration, or select an alternate method of
             calibration.

10.3    CONTINUING CALIBRATION CHECK (CCC) - An appropriate number of CCCs
       must be prepared with each extraction batch. The CCC verifies the initial calibration
       at the beginning and end of each group of analyses, and after every 101'1 sample
       during analyses. In this context, a "sample" is considered to be a field sample.
       LRBs, LFMs, LFMDs and CCCs are not counted as samples.  The beginning CCC
       for each analysis batch must be at or below the MRL in order to verify instrument
       sensitivity prior to any analyses. If standards have been prepared such that all low
       CAL points are not in the same CAL solution, it may be necessary to analyze two
       CAL solutions  to meet this requirement.  Subsequent CCCs should alternate between
       a medium and high concentration.

       10.3.1 Inject an aliquot of the appropriate concentration calibration check standard
             solution prepared with the extraction batch and analyze with the same
             conditions used during the initial calibration

       10.3.2 Calculate the concentration  of each analyte and surrogate in the check
             standard.  The calculated amount  for each analyte for medium and high level
             CCCs must be  30% of the true value.  The calculated amount for the lowest
             calibration level for each analyte must be within  50% of the true value.  If
             these conditions do not exist, then all data for the problem analyte must be
             considered invalid, and remedial action should  be taken which may require
             recalibration. Any field  sample extracts that have been analyzed since the last
             acceptable calibration verification should be reanalyzed after adequate
             calibration has been restored, with the following exception. If the continuing
             calibration fails because the calculated concentration is greater than  130%
             (150% for the low-level CCC) for a particular target compound, and field
             sample  extracts show no detection for that target compound, non-detects may
             be reported without re-analysis.
                                   515.4-22

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11.     PROCEDURE

       11.1   SAMPLE EXTRACTION AND HYDROLYSIS

             11.1.1  Remove the samples from storage (Sect 8 3) and allow them to equilibrate to
                    room temperature.

             11.1.2  Place 40 mL of the water sample into a precleaned 60-mL. glass vial with a
                    PTFE lined screw cap using a graduated cylinder.

             11.1.3  Add 10 uL of surrogate standard (lOOug/mL, 2,4-dichlorophenylacetic acid in
                    acetone per Section 7.2.2) to the aqueous sample.

             11.1.4  Add 1 mL, of the 4 N NaOH solution prepared in Section 7.1  10 to each glass
                    vial. Check the pH of the sample with pH paper or a pH meter. If the sample
                    does not have a pH greater than or equal to 12, adjust the pH by adding more
                    4 N NaOH solution.  Let the sample sit at room temperature for 1 hour,
                    shaking the contents periodically.

                    NOTE: Since many of the herbicides contained in this method are applied as a
                    variety of esters and  salts, it is vital  to hydrolyze them to the parent acid prior
                    to extraction. This step must be included in the analysis of all extracted field
                    samples, LRBs, LFMs and calibration standards.  Failure to perform this step
                    may result in data that are biased low for some targets in field samples.(8)

             11.1.5  Following hydrolysis, add 5 mL of (90:10, v:v) hexane:MlBE (Section 7.1.5)
                    and shake vigorously for three minutes.  Allow the phases to separate for
                    approximately  5 minutes then remove and discard the top hexane/MtBE layer.
                    This wash aids in sample cleanup and removes any Dacthal from the sample
                    which would interfere with  the quantitation of the Dacthal metabolites.

             11.1.6  Adjust the pH to approximately 1 by adding concentrated sulfunc acid. Cap,
                    shake and then check the pH with a pH meter or narrow range pH paper. Add
                    additional sulfuric acid as needed to properly adjust the pH.

             11.1.7  Quickly add  approximately 2 g of copper II sulfate pentahydrate and shake
                    until dissolved. This colors the aqueous phase blue and allows the analyst to
                    better distinguish between the aqueous phase and the organic phase in this
                    micro extraction.

             11.1.8  Quickly add  approximately 16 g of muffled sodium sulfate  (Sect. 7.1.7) and
                    shake until almost all is dissolved.  Sodium sulfate is added to increase the
                    ionic strength of the  aqueous phase and thus further drive the chlorophenoxy

                                         515.4-23

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             acids into the organic phase. The addition of salt also decreases the solubility
             of MtBE in the aqueous phase and allows greater volumetric recovery. The
             addition of this salt and the copper II sulfaie pentahydrate should be done
             quickly so that the heat generated from the addition of the acid (Section
             11.1.6) will help dissolve the salts.

       11.1.9 Add exactly 4.0-mL MtBE and shake vigorously for three minutes.

       11.1.10 Allow the phases to separate for approximately 5 minutes.

11.2    SAMPLE METHYLATION WITH DIAZOMETHANE

       11.2.1 GENERATION OF DIAZOMETHANE

             11.2.1.1   Assemble the diazomethane generator (Figure 4) m a hood. The
                       collection vessel is a 10- or 15-mL, glass vial equipped with a
                       PTFE lined screw cap that is maintained at 0 - 5 C.

             11.2.1.2   Add a sufficient amount of MtBE (approximately 7 mL) to tube 1
                       to cover the first impinger.  Add 10 mL of MtBE to the collection
                       vial.  Set the nitrogen flow at 5-10 mL/min.  Add 4-mL Diazald
                       solution (Sect.  7.1.13) and 3 mL of 37% KOH solution (Sect.
                       7.1.11) to the second impinger.  Connect the tubing as shown and
                       allow the nitrogen flow to purge the diazomethane from the"
                       reaction vessel into the collection vial for 30 minutes. Cap the vial
                       when collection is complete and maintain at 0-5 C  When stored
                       at 0-5 C, this diazomethane solution may be used over a period of
                       72 hours.

             11.2.1.3    Several commercial sources of glassware are available for
                       diazomethane generation.  These include a mini  Diazald apparatus
                       from Aldrich (Cat. #:Z 10,889-8).

       11.2.2 Using a Pasteur pipette, transfer the sample extract (upper MtBE layer) to a 7-
             mL, screw cap vial.  Add 0.6 g acidified sodium sulfate (Sect. 7 1 8) and
             shake.  This step is included to dry the MtBE extract.-

       11.2.3 Using a Pasteur pipette, transfer the extract to a second, 7-mL glass vial.

       11.2.4 Add 250 uL of the diazomethane solution  prepared  in Section 11.2.1 to each
             vial  The contents of the vial should remain slightly yellow in color indicating
             an excess of diazomethane. Additional diazomethane may  be added if
             necessary.  Let the estenfication reaction proceed for 30 minutes.

                                  515.4-24

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       11.2.5 Remove any unreacted diazomethane by adding 0.1 g of si lica gel.
             Effervescence (evolution of nitrogen) is an indication that excess
             diazomethane was present. Allow the extracts to sit for 0.5 hour.

       11.2.6 Transfer the extract to an autosampler vial  A duplicate vial may be filled
             using the excess extract.

       11.2.7 Analyze the sample extracts as soon as possible.  The sample extract may be
             stored up to 21 days if kept at 0 C or less. Keep the extracts away from light
             in amber glass vials with PTFE lined caps.

11.3    GAS CHROMATOGRAPHY

       11.3.1 If the fast GC option is used, several important changes from "conventional
             GC" should be made to aid in the rapid analysis of the analytes  The
             instrument should have a fast temperature ramp (50 C/mmute) oven and a
             high pressure (> 50 psi) split/splitless injector. When using columns with
             diameters less than 180 urn the instrument should be equipped with a low
             volume (150 |jL) micro ECD. Additionally, the column diameter and film
             thickness should be decreased and the carrier gas should be changed to
             hydrogen.

       11.3.2 Use of Hydrogen Safely - Although hydrogen can be used safely as a carrier
             gas, the potential for fire or explosion does exist if the gas system is
             mishandled. If you are unsure of the safety guidelines for using hydrogen as a
             carrier gas,  seek advice from your instrument manufacturer regarding its use.

       11.3.3 Column Selection and  Installation - During method development the RTX-
             1701 columns provided baseline  resolution for the 2,4-DB /Chloramben pair
             (peaks 11 and 12 in Figure  1).  Columns from other manufacturers were not as
             effective in separating these analytes. Strict attention must be paid to
             established  column installation guidelines with regard to the proper cutting
             and placement of the capillary columns within the instrument. Additionally, a
             loss of response over time was noted for Acifluorifen and Dmoseb.  Trimming
             approximately 1 meter from the head of the column restored the response for
             these analytes. If conditions in the laboratory necessitate frequent column
             trimming, a guard column is recommended.

11.4    ANALYSIS OF EXTRACTS

       11.4.1 Establish operating conditions as described in Section 17, Table 1 (Table 2 if
             performing confirmation).  Confirm that retention times, compound separation
             and resolution are similar to those summarized m Table 1 and Figure 1.

                                  515.4-25

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              11.4.2 Establish an appropriate retention time window for each target and surrogate
                    to identify them in the QC and field samples. This should be based on
                    measurements of actual retention time variation for each compound in
                    standard solutions analyzed on the GC over the course of time.  Plus or minus
                    three times the standard deviation of the retention time for each compound
                    while establishing the initial calibration and completing the IDC can be used
                    to calculate a suggested window size; however, the experience of the analyst
                    should weigh heavily on the determination of the appropriate retention
                    window size.

              11.4.3 Check system calibration by analyzing a CCC (Sect. 10.3) and calibrate the
                    system by either the analysis of a calibration curve (Sect. 10. 2) or by
                    confirming the initial calibration is still valid by analyzing a continuing
                    calibration check as described in Section 10.3. Begin analyzing field and QC
                    samples at their appropriate frequency by injecting the same size aliquots
                    under the same conditions used to analyze the initial calibration.

              11.4.4 An analyst must not extrapolate beyond the established calibration range. If
                    an analyte result exceeds the range of the initial calibration curve, the extract
                    may be diluted with MtBE containing the internal standard (Sect. 7.2.1.3), and
                    the diluted extract injected. Acceptable surrogate performance (Sect. 9.8)
                    should be determined from the undiluted sample extract.  Incorporate the
                    dilution factor into final concentration calculations. The dilution will also
                    affect analyte MRLs.
12.    DATA ANALYSIS AND CALCULATION

       12.1    Identify the method analytes in the sample chromatogram by comparing the retention
              time of the suspect peak to retention time of an analyte peak in a calibration standard.
              Surrogate retention times should be confirmed to be within acceptance limits (Sect.
              11.4.2) even if no target compounds are detected.

       12.2    Calculate the analyte concentrations using the initial calibration curve generated as
              described in Section 10.2. Quantitate only those values that fall between the MRL
              and the highest calibration standard.  Samples with target analyte responses that
              exceed the highest standard require dilution and reanalysis (Sect. 11 4.4).

       12.3    Analyte identifications should be confirmed using the confirmation column specified
              in Table 2 or another column that is dissimilar to the primary column or by GC/MS if
              the concentrations are sufficient.

       12.4    Adjust the calculated concentrations of the detected analytes to reflect the initial

                                          515.4-26

-------
             sample volume and any dilutions performed.

       12.5   Analyte concentrations are reported in ug/L as the total Tree acid.  Calculations should
             use all available digits of precision.

13.    METHOD PERFORMANCE

       13.1   PRECISION, ACCURACY, AND Detection Limits - Tables for these data are
             presented in Section 17. Single laboratory precision and accuracy data are presented
             in Tables 5 -10. Detection Limits are presented in Table 4 and  were calculated using
             the formula presented in Section 9.2.4.

14.    POLLUTION PREVENTION

       14.1   This method utilizes liquid:liquid microextraction to extract analytes from water. It
             requires the use of very small volumes of organic solvent and very small quantities of
             pure analytes, thereby minimizing the potential hazards to both  the analyst and the
             environment as compared to the use of large volumes of organic solvents in
             conventional liquid-liquid extractions.

       14.2   For information about pollution prevention that may be applicable to laboratory
             operations, consult "Less is Better: Laboratory Chemical Management for Waste
             Reduction" available from the American Chemical Society's Department of
             Government Relations and Science Policy, 1155 16th Street N.W., Washington,  D.C.,
             20036.

15.    WASTE MANAGEMENT

       15.1   The analytical procedures described in this method generate relatively small amounts
             of waste  since only small amounts of reagents and solvents are used  The matrices of
             concern are finished drinking water or source water. However,  the Agency requires
             that laboratory waste management practices be conducted consistent with all
             applicable rules and regulations, and that laboratories protect the air, water, and land
             by minimizing and controlling all  releases from fume hoods and bench operations.
             Also, compliance is required with any sewage discharge permits and regulations,
             particularly the hazardous waste identification rules and land disposal restrictions.
             For further information on waste management, consult "The Waste Management
             Manual for Laboratory Personnel" also available from the American Chemical
             Society at the address in Section 14.2.
                                         515.4-27

-------
16.    REFERENCES

      1.     Glaser, J.A., Foerst, D.L., McKee, G.D., Quave, S.A., and Budde, W.L., "Trace
             Analyses for Wastewaters." Environ. Sci. Technol.. 15 (1981) 1426-1435.

      2.     Giam, C.S., Chan, H.S., and Nef, G.S., "Sensitive Method for Determination of
             Phthalate Ester Plasticizers in Open-Ocean Biota Samples," Analytical Chemistrv.47.
             2225 (1975).

      3.     Giam, C.S., and Chan, H.S., "Control of Blanks in the Analysis of Phthalates in Air
             and Ocean Biota Samples,"  U.S. National Bureau of Standards, Special Publication
             442, 701-708, (1976).

      4.     "OSHA Safety and Health Standards, General Industry," (29CRF1910)  Occupational
             Safety and Health Administration, OSHA 2206, (Revised, Jan. 1976).

      5.     "Carcinogens-Working with Carcinogens," Publication No. 77-206, Department of
             Health, Education, and Welfare, Public Health Service, Center for Disease Control,
             National Institute of Occupational Safety and Health, Atlanta, Georgia, August 1977.

      6.     "Safety In Academic Chemistry Laboratories," 3rd Edition, American Chemical
             Society Publication, Committee on Chemical Safety, Washington, D.C., (1979).

      7.     ASTM Annual Book of Standards, Part II, Volume 11.01, D3370-82, "Standard
             Practice for Sampling Water," American Society for Testing and Materials,
             Philadelphia, PA, 1986.

      8.     Bassett, M.V.; Pepich, B.V. and Munch, D.M., "The Role Playedby Basic Hydrolysis
             in the Validity of Acid herbicide Data - Recommendations for Future Preparation of
             Herbicide Performance Evaluation Standards." Environ.  Sci. Technol.. Vol.34, 1117,
             (2000).
                                        515.4-28

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17. TABLES. DIAGRAMS. FLOWCHARTS. AND VALIDATION DATA

TABLE 1: CHROMATOGRAPHIC CONDITIONS AND AVERAGE RETENTION TIME
DATA FOR THE PRIMARY COLUMN (RTX-1701,  40m x 180 urn i.d.)
Peak Number
(Figure 1)
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
Compound
Dalapon
3,5 Dichlorobenzoic acid
2,4-Dichlorophenylacetic acid (SUR)
Dicamba
Dichlorprop
4,4'-Dibromooctafluorobiphenyl (IS)
2,4-D
Pentachlorophenol
Silvex
2,4,5-T
2,4-DB
Chlorambcn
Dinoseb
Bentazon
Dacthal
Quinclorac
Picloram
Acifluorfcn
Average Tr
(min)a
7.06
11.32
11.80
11.91
12.21
12.25
12.37
12.49
12.70
12.90
13.10
13.15
13.44
13.60
13.68
13.85
14.02
15.20
% RSD
0.017
0.002
0.002
0.002
0.002
0.002
0.002
0.002
0.002
0.002
0.003
0.003
0.003
0.003
0.002
0.004
0.003
0004
" The average retention time represents the ax crage of 7 runs

Primary Column.
      DB-1701,40 m x 0 ISO mm i d . 0.20 urn film thickness, injector temp. 200 "C, liner 2 mm straight siltek"
      deactivated, injection volume 1 uL of 200 ng/mL (highest level component) standard, splitless injection hold to
      1 mm then purge @ 50 mL/min, detector temp 360 'C, detector make up gas nitrogen at 20 mL/mmute
      Temperature program 45 "C initial, program at 5 "C/mmute to 80 C. Then 50 "C/mmute to 220 C, Then 20
      'C/mmute to 280 'C and Hold for 3 mm. Data collection via HP GC Chemstation at a rate of 20Hz.

Carrier gas- Hydrogen (UHP)

Detector makeup gas Nitrogen (UHP)
                                        515.4-29

-------
TABLE 2: CHROMATOGRAPHIC CONDITIONS AND AVERAGE RETENTION TIME
DATA FOR THE CONFIRMATION COLUMN (DB-5, 40 m x 180 urn i.d.)
Peak Number
(Figure 1)
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
Compound
Dalapon
3,5-Dichlorobenzoic acid
2,4-Dichlorophenylacetic acid (SUR)
Dicamba
Dichlorprop
4,4'-Dibromooctafluorobiphenyl (IS)
2,4-D
Pentachlorophenol
Silvex
2,4,5-T
2,4-DB
Chloramben
Dinoseb
Bentazon
Dacthal
Quinclorac
Picloram
Acifluorfen
Average Tr
(min)'
5.04
10.33
11.31
11.46
12.19
12.71
12.34
13.16
13.29
13.48
13.94
13.34
14.01
14.28
1500
14.88
14.59
16.38
% RSD
0057
0.007
0.004
0.006
0.006
0.004
0.005
0.006
0.004
0005
0.005
0.004
0.004
0.006
0.004
0.004
0.004
0.003
 The average retention time represents the average of 7 runs.

Confirmation Column
       DB-5,40 m x 0 180 mm i.d., 0.20 urn film thickness, injector temp 200 "C, liner 2 mm stiaight siltek"
       deactivated, injection volume 1 uL of 100 ng/mL (highest level component) standard, splitless injection hold to
       1 mm then purge @ 50 mL/min, detector temp 360 'C, detector make up gas nitrogen at 20 mL/mmute
       Temperature program- 45 "C initial, program at 4 'C/mmute to 80 "C then program at 30 C/mmute to 280 and
       Hold for 2 mm, Data collection via HP GC Chemstation at a rate of 20Hz

Carrier gas. Hydrogen (UHP)

Detector makeup  gas Nitrogen (UHP)
                                          515.4-30

-------
TABLE 3: CHROMATOGRAPHIC CONDITIONS AND AVERAGE RETENTION TIME
DATA FOR A CONVENTIONAL PRIMARY COLUMN (DB-5, 30 m x 250 urn i.d.)
Peak Number
(Figure 3)
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
Compound
Dalapon
3,5-Dichlorobenzoic acid
2,4-Dichlorophenylacetic acid (SUR)
Dicamba
Dichlorprop
4,4'-Dibromooctafluorobiphenyl (IS)
2,4-D
Pentachlorophenol
Silvex
2,4,5-T
2,4-DB
Chloramben
Dinoseb
Bentazon
Dacthal
Quinclorac
Picloram
Acifluorfen
Tr
(min)
14.28
20.35
2098
21 10
21.46
21.53
21.64
21.81
22.03
22.27
22.51
22.55
22.94
23.17
23.30
23.55
23.82
25.90
Conventional Primary Column.
       DB-1701, 30 mx 0 250 mm i d., 0.25 \im film thickness, mjectoi icmp 200 "C, liner 2 mm snaight deactivated,
       injection volume 1 uL of 100 ng/mL (highest level component) standard, sphtless injection hold to 5 mm then
       purge @ 30 mL/mm, detector temp 290 "C, detector make up gas P-5 at 60 mL/mmute  Tempeiature program
       45 "C initial, hold 10 mm then program at 5 'C/mmute to 70 'C, Then 30 'C/mmute to 260 'C. and Hold for 5
       mm, Data collection via Turbochrom 4.1

Carrier gas. Helium (UHP)

Detector makeup gas  5% methane/argon (UHP)
                                         515.4-31

-------
TABLE 4: DETECTION LIMITS IN REAGENT WATER
Compound
Dalapon
3,5-Dichlorobenzoic acid
Dicamba
Dichlorprop
2,4-D
Pentachlorophenol
Silvex
2,4,5-T
2,4-DB
Chloramben
Dinoseb
Bentazon
Dacthal
Quinclorac
Picloram
Acifluorfen
Fortification
Level (ug/L)
0.100
0.050
0.050
0.100
0.100
0.010
0.025
0.025
0.100
0.050
0.100
0.100
0.050
0.050
0.050
0.250
Primary
Column
Detection
LimitJ (ug/L)
0.054
0212
0.032
0.433
0.055
0014
0.033
0.024
0.246
0.057
0.166
0.064
0.113
0.080
0.076
0.307
Secondary
Column
Detection
LimitJ (ug/L)
0074
0.049
0042
0.12
0066
0.084
0018
0.033
0.182
0.083
0.081
0.185
0105
0.113
0.055
0.092
      "Detection Limits were extracted and analyzed over 3 days for 9 replicates following
      the procedure outlined in Section 9
                                     515.4-32

-------
TABLE 5: LOW LEVEL PRECISION AND ACCURACY IN REAGENT WATER
Compound
Dalapon
3,5-Dichlorobenzoic acid
Dicamba
Dichlorprop
2,4-D
Pentachlorophenol
Silvex
2,4,5-T
2,4-DB
Chloramben
Dinoseb
Bentazon
Dacthal
Quinclorac
Picloram
Acifluorfen
2,4-Dicblorophenylacetic acid (SUR)
Fortification
Level (ug/L)
1.0
0.50
0.50
1.0
1.0
0.10
0.25
0.25
1.0
0.50
1.0
1.0
0.50
0.10
1.0
0.50
25
Mean %
Recover}'
108
117
97
95
98
76
84
96
97
90
103
100
92
106
92
77
104
% RSD
(n=7)
2.2
16
3.7
57
5.2
74
20
7.3
8.4
9.0
6.7
2.1
6.5
9.3
2.6
5.9
8.0
S/N
Ratio"
578
9
35
19
23
21
9
16
6
113
8
41
100
20
23
80
-
      aSignal-to-noise ratios were calculated for each target compound peak by dividing the
      peak height for each compound by the peak-to-peak noise, which was determined for
      each component from the method blank over a period of time equal to the full peak
      width in the target analyte's retention time window.
                                     515.4-33

-------
TABLE 6: MID LEVEL PRECISION AND ACCURACY IN REAGENT WATER
Compound
Dalapon
3,5-Dichlorobenzoic acid
Dicamba
Dichlorprop
2,4-D
Pentachlorophenol
Silvex
2,4,5-T
2,4-DB
Chloramben
Dinoseb
Bentazon
Dacthal
Quinclorac
Picloram
Acifluorfen
2,4-Dichlorophenylacetic acid (SUR)
Fortification
Level (ug/L)
10
5.0
5.0
10
10
1.0
2.5
2.5
10
5.0
10
10
5.0
1.0
10
5.0
25
Mean %
Recovery
107
96
102
107
106
103
107
105
93
105
119
98
100
101
99
107
112
% RSD
(n=7)
23
2.3
1.5
0.6
1 4
1.4
4.4
1.3
1.6
2.9
4.6
0.7
1.9
2.9
3.1
4.5
1.3
                             515.4-34

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TABLE 7: LOW LEVEL PRECISION AND ACCURACY IN FORTIFIED CHLORINATED
SURFACE WATER
Compound
Dalapon
3,5-Dichlorobenzoic acid
Dicamba
Dichlorprop
2,4-D
Pentachlorophenol
Silvex
2,4,5-T
2,4-DB
Chloramben
Dinoseb
Bentazon
Dacthal
Quinclorac
Picloram
Acifluorfen
2,4-Dichlorophenylacetic acid (SUR)
Fortification
Level (ug/L)
10
0.50
0.50
1.0
1.0
0.10
0.25
0.25
1.0
0.50
1.0
1.0
0.50
0.10
1.0
0.50
25
Mean %
Recovery
104
110
93
93
97
86
103
100
104
93
99
91
92
96
102
98
107
% RSD
(n=7)
2.1
3.0
4.4
11
5.2
3.2
4.3
3.5
5.9
2.0
1.2
2.2
4.6
15
2.4
4.1
1 5
                             515.4-35

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TABLE 8: MID LEVEL PRECISION AND ACCURACY IN FORTIFIED CHLORINATED
SURFACE WATER
Compound
Dalapon
3,5-Dichlorobenzoic acid
Dicamba
Dichlorprop
2,4-D
Pentachlorophenol
Silvex
2,4,5-T
2,4-DB
Chloramben
Dinoseb
Bentazon
Dacthal
Quinclorac
Picloram
Acifluorfen
2,4-Dichlorophenylacetic acid (SUR)
Fortification
Level (ug/L)
10
5.0
5.0
10
10
1.0
2.5
2.5
10
5.0
10
10
50
1.0
10
5.0
25
Mean %
Recovery
100
104
102
101
101
100
101
100
101
102
101
98
99
105
107
92
106
% RSD
(n=7)
2.5
2.6
29
1.2
1.2
1.0
1.0
1.6
2.3
2.6
1.6
1.8
2.0
2.5
3.2
2.4
1.4
                             515.4-36

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TABLE 9: LOW LEVEL PRECISION AND ACCURACY IN FORTIFIED CHLORINATED
GROUND WATER
Compound
Dalapon
3,5-Dichlorobenzoic acid
Dicamba
Dicblorprop
2,4-D
Pentachlorophenol
Silvex
2,4,5-T
2,4-DB
Chloramben
Dinoseb
Bentazon
Dacthal
Quinclorac
Picloram
Acifluorfen
2,4-Dichlorophenylacetic acid (SUR)
Fortification
Level (ug/L)
1.0
0.50
0.50
1.0
1.0
0.10
0.25
0.25
1.0
0.50
1.0
1.0
0.50
0.10
1.0
0.50
25
Mean %
Recovery
105
104
100
109
108
103
96
91
94
88
100
101
62a
107
97
92
95
% RSD
(n=7)
1.5
6.0
2.2
3.5
10
17
3.0
3.4
3.1
2.7
4.0
2.8
9.9
5.7
2.8
4.2
3.1
      "Dacthal was present in the chlorinated surface water at about l/5th the fortification
      level. This was taken into account in the determination of the Mean % Recovery.
                                    515.4-37

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TABLE 10: MID LEVEL PRECISION AND ACCURACY IN FORTIFIED CHLORINATED
GROUND WATER
Compound
Dalapon
3,5-Dichlorobenzoic acid
Dicamba
Dichlorprop
2,4-D
Pentachlorophenol
Silvex
2,4,5-T
2,4-DB
Chloramben
Dinoseb
Bentazon
Dacthal
Quinclorac
Picloram
Acifluorfen
2,4-Dichlorophenylacetic acid (SUR)
Fortification
Level (ug/L)
10
5.0
5.0
10
10
1.0
2.5
2.5
10
5.0
10
10
5.0
1.0
10
5.0
25
Mean %
Recovery
97
95
98
79
99
100
97
97
92
102
105
101
99
94
103
84
106
% RSD
(n=7)
5.5
2.1
1.6
0.7
1.6
4.2
1.6
1.5
1.1
2.6
2.6
1.8
1.2
1.5
2.5
2.5
1.1
                             515.4-38

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TABLE 11.  SAMPLE HOLDING TIME DATA FOR SAMPLES FROM A
           CHLORINATED SURFACE WATER FORTIFIED WITH METHOD
           ANALYTES"
Compound
Dalapon
3,5-Dichlorobenzoic acid
Dicamba
Dichlorprop
2,4-D
Pentachlorophenol
Silvex
2,4,5-T
2,4-DB
Chloramben
Dinoseb
Bentazon
Dacthal
Quinclorac
Picloram
Acifluorfen
DayO
% Rec
91
98
92
89
88
92
91
89
87
86
105
91
92
90
94
109
Day 2
% Rec
90
97
92
88
85
85
88
89
84
86
95
87
91
80
85
96
Day 7
% Rec
91
85
96
91
87
86
86
85
86
88
99
95
86
84
85
103
Day 14
% Rec
108
91
99
95
92
93
94
91
93
100
111
99
94
91
99
123
     a All samples fortified at the same level used to collect the mid level precision and
     accuracy data (see Table 10 for example).
                                  515.4-39

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TABLE 12.   EXTRACT HOLDING TIME DATA FOR SAMPLES FROM A
            CHLORINATED SURFACE WATER FORTIFIED WITH METHOD
            ANALYTES"1"
Compound
Dalapon
3,5-Dichlorobenzoic acid
Dicamba
Dichlorprop
2,4-D
Pentach lorophenol
Silvex
2,4,5-T
2,4-DB
Chloramben
Dinoseb
Bentazon
Dacthal
Quinclorac
Picloram
Acifluorfen
2,4-Dichlorophenylacetic acid
(SUR)
Initial a b
Injection
%Rec
100
104
102
100
101
100
101
99
101
102
99
98
99
105
107
92
106
Day 21
Reinfection
%Rec
89
113
104
101
102
102
103
102
102
105
81
100
103
127
98
104
105
      "Sample storage stability is expressed as a percent recovery (%Rec) value calculated as
      described in Section 9.9.2.

      bAll samples fortified at the same level used to collect the mid level precision and
      accuracy data (see Table 10 for example).
                                    515.4-40

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TABLE 13: INITIAL DEMONSTRATION OF CAPABILITY (IDC) REQUIREMENTS
Method
Reference
Section
9.2.1







Section
9.2.2

Section
9.2.3

Section
9.2.4










Requirement

Initial
Demonstration of
Low Method
Background





Initial
Demonstration of
Precision (IDP)
Initial
Demonstration of
Accuracy
Detection Limit
Determination










Specification and
Frequency
Analyze LRB prior to any
other IDC steps







Analyze 4-7 replicate LFBs
fortified at midrange
concentration.
Calculate average recovery
for replicates used in IDP

Over a period of three days,
prepare a minimum of 7
replicate LFBs fortified at a
concentration estimated to
be near the Detection Limit.
Analyze the replicates
through all steps of the
analysis. Calculate the
Detection Limit using the
equation in Section 9.2.4.


Acceptance Criteria

Demonstrate that all target
analytes are below V3 the
reporting limit or lowest CAL
standard, and that possible
interference from extraction
media do nol prevent the
identification and
quantification of method
analytes.
%RSDmustbe <;20%


Mean recovery must be within
20% of true value.

Must be <, 1/3 of the
established MRL

Note: Data from Detection
Limit replicates are not
required to meet method
precision and accuracy criteria.
If the Detection Limit
replicates are fortified at a low
enough concentration, it is
likely that they will not meet
precision and accuracy criteria.
                              515.4-41

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TABLE 14: QUALITY CONTROL REQUIREMENTS (SUMMARY)
 Method
 Reference
Requirement
Specification and
Frequency
Acceptance Criteria
 Section
 10.2
Initial Calibration
Use internal standard
calibration technique to
generate a calibration curve.
Use at least 5 standard
concentrations.
When each calibration standard
is calculated as an unknown
using the calibration curve, the
result must be 70-130% of the
true value for all except the
lowest standard, which must be
50-150% of the true value.
 Section
 9.4
Laboratory
Reagent Blank
(LRB)
Daily, or with each
extraction batch of up to 20
samples, whichever is more
frequent.
Demonstrate that all target
analytes are below V3 the
method reporting limit or
lowest CAL standard, and that
possible interferences do not
prevent quantification of
method analytes.
If targets exceed '/, the MRL,
results for all subject analytes
in extraction batch are invalid.
 Sections
 9.5 and
  10.3
Continuing
Calibration Check
(CCC)
Verify initial calibration by
analyzing a calibration
standard at the beginning of
each analysis batch prior to
analyzing samples, after
every 10 samples, and after
the last sample.

Low CCC - near MRL
Mid CCC - near midpoint in
initial calibration curve
High CCC - near highest
calibration standard
1) The result for each analyte
must be 70-130% of the true
value for all but the lowest
standard. The lowest standard
must be 50-150% of the true
value.
2) The peak area of internal
standards must be 50-150% of
the average peak area
calculated during the initial
calibration.
Results for  analytes that do not
meet IS criteria or are not
bracketed by acceptable CCCs
are invalid.
                                         515.4-42

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TABLE 14: QUALITY CONTROL REQUIREMENTS (SUMMARY)
Method
Reference
Section 9.7






Section 9.8




Section 9.9










Section
9.10






Section
9.11


Requirement

Internal
Standard
(IS)




Surrogate
Standard
(SUR)


Laboratory
Fortified
Sample
Matrix (LFM)
and
Laboratory
Fortified
Matrix
Duplicate
(LFMD)

Field
Duplicates
(FD)





Quality
Control
Sample
(QCS)
Specification and
Frequency
4,4'-dibromooctafluoro-
biphenyl is added to the
extraction solvent.




2,4-dichlorophenylacetic
acid is added to all
calibration standards and
samples, including QC
samples.
Analyze one LFM per
analysis batch (20
samples or less) fortified
with method analytes at a
concentration close to but
greater than the native
concentration.
LFMD should be used in
place of Field Duplicate if
frequency of detects for
targets is low.
Extract and analyze at
least one FD with each
extraction batch (20
samples or less). A
LFMD may be substituted
for a FD when the
frequency of detects for
target analytes is low.
Analyzed when new
Primary Dilution
Standards (PDS) are
prepared.
Acceptance Criteria

Peak area counts for the IS m LFBs,
LRBs, and sample extracts must be
within 50-150% of the average peak
area calculated during the initial
calibration. If the IS does not meet
criteria, corresponding target results
are invalid.
Surrogate recovery must be 70-130%
of the true value. If surrogate fails
this criterion, report all results for
sample as suspect/surrogate recovery.

Recoveries at mid and high levels not
within 70-130% or low-level
recoveries not within 50-1 50% of the
fortified amount may indicate a matrix
effect.
Target analyte RPDs for LFMD
should be within 30% at mid and
high levels of fortification and within
50% near MRL.


Target analyte RPDs for FD should be
within 30% at mid and high levels of
fortification and within 50% near
MRL.




Results must be 70-130% of the
expected value.


                              515.4-43

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Method
Reference
Requirement
Specification and
Frequency
Acceptance Criteria
Section 8.4
Sample
Holding Time
14 days with appropriate
preservation and storage
Sample results are valid only if
samples are extracted within sample
hold time
Section 8.4
Extract
Holding Time
 21 days with appropriate
storage
Sample results are valid only if
extracts are analyzed within extract
hold time.
                                        515.4-44

-------
                            FIGURE 1
CHROMATOGRAM OF THE CHLORPHENOXY HERBICIDES ON A RTX-1701 COLUMN
         CHROMATOGRAPHIC CONDITIONS ARE GIVEN IN TABLE 1.
Hz
22500-
20000
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17500-

15000
.
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-
7500 
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                                                                  18
                                                                15 min
                            515.4-45

-------
                                                   FIGURE 2
                   CHROMATOGRAM OF THE CHLORPHENOXY HERBICIDES ON A DB-5 COLUMN
                            CHROMATOGRAPHIC CONDITIONS ARE GIVEN IN TABLE 2.
  Hz

10000
 8000
6000
4000
2000
                      Vl
ATh"-ufiv"7' ",', Avivv >-"i'i-,
                                                  10                 12
                                                                                12
                                                                              89
                                                                                 10
                                                                                      13
              17
                                                                                               15
                                                                                             16
                                                                                        14
W V* I- V; h H," n'S I,' ' ifr-'-f
                                                                                     14
                                                                                                           IS
                           I
                          16   min
                                                   515.4-46

-------
                                             FIGURE 3
       CHROMATOGRAM OF THE CHLORPHENOXY HERBICIDES ON A CONVENTIONAL PRIMARY COLUMN
                  (30 m x 250 urn) CHROMATOGRAPHIC CONDITIONS ARE GIVEN IN TABLE 3.
1 2O
1OQ
 8 fl-
-40
                          VA
                                                                       13
                                                                 10
                                                                \J
                                                                   12
                                                                  II
                                                                             17
                                                                         15
                                                                        14
                                                                           16
          'I1111!1111!1
          15      16
I1 '"I
                                                 18
                         17
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 19
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11111111 1111 [ I
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I I I I | I I I I | I
  22
rrp-
 23
I I I I | I I I I | I I I I | I
  2^      25
I11"!11
   26
                                             5 I 5.4-47

-------
                               FIGURE 4
                      DIAZOMETHANE GENERATOR
   Nitrogen
MtBE
        Diazald+KOH
                                                        MtBE in collection
                                                              vial

                                                         Ice bath
                                515.4-48

-------