:WeJtfto:dJ5133  Determination of Chlorinated Herbicides
                   1n Drinking Hater
                    September 1986

     Supplement to "Methods for the Determination
       of  Organic  Compounds 1n Finished Drinking
              Hater and Raw Source Water"
          Physical  and Chemical  Methods  Branch
    Environmental Monitoring and Support Laboratory
         U. S. Environmental Protection Agency
                 Cincinnati,  Ohio 45268

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           METHOD 515:  THE DETERMINATION OF CHLORINATED HERBICIDES
                               IN  DRINKING  WATER
1.  SCOPE AND APPLICATION"

    1.1  This method is applicable to the determination of certain
         chlorinated acid herbicides in drinking water.  This method is
         applicable, but not limited to, the analyses of the following
         parent acids and salts and esters of these acids.  The chemical
         form of each acid is not distinguished.  Results are calculated and
         reported as total  free acid.
     »

                                               Chemical Abstract Services
              Analyte                          	Registry Number (CAS)

         2,4-D                                           94-75-7
         2,4-08                                          94-82-6
         Dicamba                                       1918-00-9
         2,4,5-T                                         93-76-5
         2,4,5-TP (Silvex)                                93-72-1
         Dalapon                          •               75-99-0
         Pentachlorophenol  (PCP)                         87-86-5
         Dinoseb                                         88-85-7
         Picloram                                      1918-02-1

    1.2  The estimated detection limit (EDL) for each analyte above is given
         in Table 1 for capillary column chromatography.  The EDLs were
         determined by the esterification and analyses of replicate 1 mL
         acid calibration standards as described in Sect. 7.3.   Thus the
         variability represented by the EDL  is that of the esterification
         and analysis portions and not of the total procedure.   The method
         detection limit (MDL) for each analyte is given in Table 2 for
         packed column chromatography.

    1.3  This method is restricted to use by or under the supervision of
         analysts experienced in the use of  GC and in the interpretation of
         gas chromatograms.  Each analyst must demonstrate the  ability to
         generate acceptable results with this method using the procedure
         described in Section 8.3.

    1.4  When this method is used to analyze unfamiliar samples for any or
         all of the analytes above, identifications should be supported by
         at least one additional qualitative technique.

2.  SUMMARY OF METHOD

    2.1  A measured volume of sample, ca. 1  liter, is acidified to convert
         any salts present to the parent acid.  The acids and esters are
         extracted with ethyl ether.  The esters are hydrolyzed and

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         converted to acid salts with potassium hydroxide solution. The
         aqueous phase containing the add salts 1s then solvent washed to
         remove extraneou-s organic material.  After acidification the acids
         are extracted into organic phase and the sample volume reduced to 5
         ml in methyl-t-butyl ether (MTBE) with a K-D concentrator.  The
         acids are converted to their methyl esters by the use of a
         micro-diazomethane generator.  The samples are then analyzed by
         either packed or capillary column GC using an electron capture
         detector (ECD).l

    2.2  This method is a modified version of EPA Method 615, "The
         Determination of Chlorinated Herbicides in Industrial and Municipal
         Wastewater."

3.  INTERFERENCES

    3.1  Method interferences may be caused by contaminants in solvents,
         reagents, glassware and other sample processing apparatus that lead
         to discrete artifacts or elevated baselines in gas chromatograms.
         All reagents and apparatus must be routinely demonstrated to be
         free from interferences under the conditions of the analysis by
         running laboratory reagent blanks as described in Section 8.6.

         3.1.1  Glassware must be scrupulously cleaned.2  Clean all glass-
                ware as soon as possible after use by thoroughly rinsing
                with the last solvent used in it.  Follow by washing with
                hot water and detergent and thoroughly rinsing with dilute
                acid, tap and reagent water.  Drain dry, and heat in an oven
                or muffle furnace at 400*C for 15 to 30 min.  Do not heat
                volumetric ware.  Thermally stable material, such as PCBs,
                might not be eliminated by this treatment.  Thorough rinsing
                with acetone and pesticide quality hexane may be substituted
                for the heating.  After drying and cooling, seal and store
                glassware in a clean environment to prevent any accumulation
                of dust or other contaminants.  Store inverted or capped
                with aluminum foil.

         3.1.2  The use of high purity reagents and solvents helps to
                minimize interference problems.  Purification of solvents by
                distillation in all-glass systems may be required.

    3.2  The acid forms of the herbicides are strong organic acids, which
         react readily with alkaline substances and can be lost during
         analysis.  Glassware and glass wool must be acid-rinsed with (1+9)
         hydrochloric acid and the sodium sulfate must be acidified with
         sulfuric acid prior to use to avoid this possibility (See Section
         6.5).'

    3.3  Organic acids and phenols, especially chlorinated compounds, cause
         the most direct interference with the determination.  Alkaline
         hydrolysis and subsequent extraction of the basic solution remove
         many chlorinated hydrocarbons and phthalate esters that might
         otherwise interfere with the electron capture analysis.

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    3.4  Matrix Interferences may be caused by contaminants that are
         coextracted from the sample.  The extent of matrix Interferences
         may vary from source to source, depending upon the municipality
         sampled.  Specific cleanup procedures have not yet been identified
         for drinking water samples.

4.  SAFETY

    4.1  The toxicity or carcinogenitity of each reagent used in this method
         has not been precisely defined; however, each chemical compound
         must be treated as a potential health hazard.  From this viewpoint,
         exposure to these chemicals must be reduced to the lowest possible
         level by whatever means available.  The laboratory is responsible
         for maintaining a current awareness file of OSHA regulations
         regarding the safe handling of the chemicals specified in this
         method.  A reference file of material data handling sheets should
         also be made available to all personnel involved in the chemical
         analysis.  Additional references to laboratory safety are available
         and have been identified3-5 for the information of the analyst.

    4.2  Diazomethane is a toxic carcinogen and can explode under certain
         conditions.  The following precautions must be followed:

         4.2.1  Use only in a well ventilated hood - do not breath vapors.

         4.2.2  Use a safety screen.

         4,2.3  Use mechanical pipetting aides.

         4.2.4  Do not heat above 90*C - EXPLOSION may result.

         4.2.5  Avoid grinding surfaces, ground glass joints, sleeve
                bearings, glass stirrers - EXPLOSION may result.

         4.2.6  Store away from alkali metals - EXPLOSION may result.

         4.2.7  Solutions of dlazomethane decompose rapidly in the presence
                of solid materials, such as copper powder, calcium chloride,
                and boiling chips.

5.  APPARATUS AND MATERIALS

    5.1  Sample bottle - Amber borosilicate or flint glass, 1-liter or
         1-quart volume, fitted with screw caps lined with TFE-fluorocarbon
         or aluminum foil.  If amber bottles are not available, protect
         samples from light.  The container ..and cap liner must be washed,
         rinsed.with acetone or methylene chloride, and dried before use to
         minimize contamination.

    5.2  Glassware (All specifications are suggested.  Catalog numbers are
         included for illustration only).

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     5.2.1  Separatory funnels - 60-mL and 2000-
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         5.9.1  Column 1 - Capillary, DB-1, 30 m x 0.32 mm ID, 0.25 urn  film
                thickness or equivalent.

         5.9.2  Column 2 - Capillary, DB-5, 30m x 0.32 mm ID, 0.25 urn film
                thickness'or equivalent.

         5.9.3  Column 3 - 180 cm long x 4 mm ID glass, packed with 1.5X
                SP-2250/1.95* SP-2401 on Supelcoport (100/120 mesh) or
                equivalent.

         5.9.4  Column 4 - 180 cm long x 4 mm ID glass, packed with 5%
                OV-210  on Gas Chrom Q (100/120 mesh) or equivalent.

         5.9.5  Detector - Electron capture.  This detector has proven
                effective in the analysis of drinking water and was used to
                develop the method performance statements in Section 15.
                Alternative detectors, including a mass spectrometer, may be
                used in accordance with the provisions described in Section
                12.1.

6.  REAGENTS

    6.1  Reagent water - Reagent water is defined as a water in which an
         interferent is not observed at the method detection limit of each
         parameter of interest.

    6.2  Methanol - Pesticide quality or equivalent.

    6.3  Ethyl ether - (Burdick and Jackson Product No. 107 or equivalent).
         Nanograde, redistilled in glass if necessary.  Ethers must be free
         of peroxides as Indicated by EM Quant test strips •  Procedures
         recommended for removal of peroxides are provided with the test
         strips.  Ethers must be periodically tested (monthly) for peroxide
         formation during use.

    6.4  Methyl-t-butyl ether - (Burdick and Jackson Product No. 242 or
         equivalent).  Nanograde, redistilled in glass if necessary   Same
         peroxide precautions apply as for ethyl ether.

    6.5  Sodium sulfate - (ACS) Granular, acidified, anhydrous.  Heat treat
         in a shallow tray at 400*C for a minimum of 4 h to remove
         phthalates and other interfering organic substances.  Alterna-
         tively, Soxhlet extract with methylene chloride for 48 h.  Acidify
         by slurrying 100 g sodium sulfate with enough ethyl  ether to just
         cover the solid.  Add 0.1 ml concentrated sulfuric acid and mix
         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 mix-ture.  It must be below pH 4.  Store at 130*C.

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6.6  Hydrochloric acid (1+9) -  (ACS)  Add  one volume  of cone. HC1  to  9
     volumes reagent water.

6.8  Potassium hydroxide solution - 37% aqueous  solution  (W:V).
     Dissolve 37 g ACS grade KOH pellets  1n reagent  water and dilute  to
     100 ml.

6.9  Sulfuric acid solution (1+1) - Slowly add 50 ml H2$04 (sp. gr.
     1.84) to 50 ml of reagent water.

6.10 Sulfuric acid solution (1+3) - Slowly add 25 ml ^504 (sp. gr.
     1.84) to 75 ml of reagent water.  Store and maintain at 4*C.

6.11 Carbitol - (Diethylene glycol monoethyl ether), ACS.  Available
     from Aldrich Chemical Co.

6.12 Diazald - (N-methyl-N-nitroso-p-toluenesulfonamide), ACS.
     Available from Aldrich Chemical  Co.

6.13 Diazald Solution - Prepare a solution containing 10 grams Diazald
     in 100 ml of a 50:50 by volume mixture of ethyl ether and
     carbitol.  This solution is stable for one month or longer when
     stored at 4*C in an amber colored bottle with a teflon-lined screw
     cap.

6.14 Silicic acid - Chromatographic grade, nominal 100 mesh.   Store at
     130'C.

6.15 Bo.iling chips - Approximately 10/40 mesh.   Heat at 400*C for 1 h or
     Soxhlet extract with methylene chloride.

6.16 Stock standard solutions (1.00 ug/uU - Stock standard solutions
     can be prepared from pure standard materials or purchased as
     certified solutions.

     6.16.1 Prepare stock standard solutions by accurately weighing
            about 0.0100 grams of pure acids.   Dissolve the material   in
            pesticide quality MTBE and dilute to volume in a 10-mL
            volumetric flask.  Larger volumes  can be used at the con-
            venience of the analyst.  If compound purity is certified at
            96X or greater, the weight can be  used without correction to
            calculate the concentration of the  stock standard.
            Commercially prepared stock standards can be used at any
            concentration if they are certified by the manufacturer or
            by an independent source.

     6.16.2 Transfer the stock standard solutions into Telfon-sealed
            screw-cap vials.  Store at 4?C and  protect from light.
            Stock standard solutions should be  checked frequently for
            signs of degradation or evaporation,  especially just prior
            to preparing calibration standards  from them.

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         6.16.3 Secondary dilution  standard - Use stock standard solutions
                to prepare  secondary dilution standard solutions that  .
                contain the analytes 1n methyl-t-butyl ether.  The secondary
                dilution standard should be prepared at a concentration such
                that 50 to  200  uL of the solution can be added to 25, 50, or
                100 ml of WTBE  to prepare calibration solutions that bracket
                the working concentration range.

7.  CALIBRATION

    7.1  Establish GC operating parameters equivalent to those indicated in
         Tables 2 and 3.

    7.2  For each analyte of interest, prepare acid calibration standards at
         a minimum of three concentration levels by adding accurately
         measured volumes of one or more secondary dilution standards to a
         volumetric flask and diluting to volume with MTBE.  One of the
         external standards  should be representative of a concentration
         near, but greater  than the detection limits.  The other
         concentrations should corres- pond to the range of concentrations
         expected in the sample concentrates or should define the working
         range of the detector.

    7.3  Prepare methyl ester calibration standards by esterification of
         1.00 ml volumes of  the working standards as described in Section
         11.  Inject 1 to 2  wL of each calibration standard and tabulate
         peak height or area  responses against the mass of free acid
         represented by the  injection.  The results can be used to prepare a
         calibration curve  for each parameter.  Alternatively, the ratio of
         the response to the mass injected, defined as the calibration
         factor (CF), can be  calculated for each parameter at each standard
         concentra- tion.   If the relative standard deviation of the
         calibration factor  is less than 10% over the working range, the
         average calibration  factor can be used in place of a calibration
         curve.

    7.4  The working calibration curve or calibration factor must be
         verified on each working shift by the preparation of one or more
         calibration standards.  If the response for any analyte varies from
         the predicted response by more than *10%, the test must be repeated
         using a fresh calibration standard.  Alternatively, a new
         calibration curve or calibration factor must be prepared for that
         parameter.

8.  QUALITY CONTROL

    8.1  Each laboratory using this method is required to operate a formal
         quality control (QC)program.  The minimum requirements of this
         program consist of  an initial  demonstration of laboratory capa-
         bility and the analysis of spiked samples as a continuing check on
         performance.  The  laboratory is required to maintain performance
         records to define  the quality of data thus generated.  Specific
         minimum QC requirements consist of:

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     8.1.1  Demonstration of the ability to generate acceptable accuracy
            and precision with this method (Section 8.3).

     8.1.2  Definitien of method performance criteria for each spike
            concentration and analyte being measured (Section 8.4).

     8.1.3  Demonstration of continuing laboratory performance by
            monitoring analyte recoveries from spiked samples
            -(Section 8.5).

     8.1.4  Analysis of reagent blanks to detect introduction of reagent
            and glassware interferences (Section 8.6).

     8.1.5  Confirmation of detected analytes (Section 8.7).

     8.1.6  Additional quality assurance (QA) procedures as required
            (Section 8.7).

8.2  In recognition of the rapid advances occurring in chromatography,
     the analyst is permitted certain options to improve the separations
     or lower the cost of measurements.  Each time such modifications to
     the method are made, the analyst is required to repeat the
     procedure in Section 8.3.

8.3  Accuracy and Precision - To establish the ability to generate
     acceptable accuracy and precision, the analyst must perform the
     following operations.

     8.3.1  For each compound to be measured, using stock standards,
            prepare a QC check sample concentrate 1n methanol at a
            concentration of 5 ug/mL.

     8.3.2  Using a pipet, add 1.00 ml of the check sample concentrate
            to each of a minimum of four 1000-mL aliquots of reagent
            water.  Analyze the aliquots according to the method
            beginning in Section 10.

     8.3.3  Calculate the average percent recovery (R), and the standard
            deviation of the percent recovery (s), for the results.

     8.3.4  Using the appropriate data from Table 2, determine the
            recovery and single operator precision expected for the
            method, and compare these results to the values calculated
            in Section 8.3.3.  If the data are not comparable, review
            potential problem areas and repeat the test.

8.4  Method Performance Criteria - The analyst must calculate method
     performance criteria and define the performance of the laboratory
     for each spike concentration and parameter being measured.

     8.4.1  Calculate upper and lower control limits for method
            performance as follows:

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                   Upper Control  Limit  (UCL) - R + 2 s
                   Lower Control  Limit  (LCL) - R - 2 s

                where R and  s  are calculated as 1n Section 8.3.3.  The UCL
                and LCL can  be used to  construct control charts^ that are
                useful in observing trends 1n performance.

         8.4.2  The laboratory must develop and maintain separate accuracy
                statements of  laboratory performance for drinking water
                samples.  An accuracy statement for the method  is defined as
                R * s.  The  accuracy statement should be developed by the
                analysis of  four  aliquots of drinking water as  described in
                Section 8.3.2.  followed by the calculation of R and s.
                Alternatively,  the analyst may use four drinking water data
                points gathered through the requirement for continuing
                quality control in Section 8.5.  The accuracy statements
                should be updated regularly.6

    8.5  Analyte Recoveries -  The laboratory is required to collect in
         duplicate a portion of their samples to monitor spike  recoveries.
         The frequency of spiked  sample analysis must be at least 10% of all
         samples or one spiked  sample per month, whichever is greater.  One
         aliquot of the sample must be  spiked and analyzed as described in
         Section 8.3.  If the  recovery  for a particular parameter does not
         fall within the control  limits for method performance, the results
         reported for that parameter in all samples processed as part of the
         same set must be qualified as described in Section 13.3.  The
         laboratory should monitor the frequency of data so qualified to
         ensure that it remains at or below 5%.

    8.6  Reagent Blanks - Before processing any samples, the analyst must
         demonstrate through the analysis of a 1-liter aliquot of reagent
         water that all glassware and reagents interferences are under
         control.  Each time a set of samples is extracted or there is a
         change in reagents, a  laboratory reagent blank must be processed as
         a safeguard against laboratory contamination.

    8.7  Additional QC - It  is recommended that the laboratory adopt
         additional quality  assurance practices for use with this method.
         The specific practices that are more productive depend upon the
         needs of the laboratory and the nature of the samples.  Field
         duplicates may be analyzed to monitor the precision of the sampling
         technique.  When doubt exists over the identification of a peak on
         the chromatogram, confirmatory techniques such as GC with a dis-
         similar column, specific element detector,  or mass spectrometer
         must be used.  Whenever possible, the laboratory should perform
         analysis of QC materials and participate in relevant performance
         evaluation studies.

9.  SAMPLE COLLECTION, PRESERVATION, AND HANDLING

    9.1  Grab samples must be collected in glass containers.   Conventional
         sampling practices? should be followed; however, the bottle must
         not be prerinsed with sample before collection.

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    9.2  The samples must be Iced or refrigerated at 4*C from the time of
         collection until extraction.

    9.3  All samples must be extracted within seven days, and completely
         analyzed within 40 days of extraction.

10. SAMPLE EXTRACTION, HYDROLYSIS AND CONCENTRATION

   10.1  Mark the water meniscus on the side of the sample bottle for later
         determination of sample volume.  Pour the entire sample into a
         2-liter separatory funnel.  Check the pH with wide-range pH paper
         and adjust to pH less than 2 with sulfuric acid (1+1).   Determine
         the original sample volume by refilling the sample bottle to the
         mark and transferring the water to a 1000-mL graduated  cylinder.
         Record the sample volume to the nearest 5 ml.  Alternatively, the
         sample volume may be determined by weighing before and  after the
         sample bottle is emptied.

   10.2  Add 150 at ethyl ether to the sample bottle, cap the bottle and
         shake 30 s to rinse the walls.  Transfer the solvent to the
         separatory funnel and extract the sample by shaking the funnel  for
         2 min with periodic venting to release excess pressure.  Allow the
         organic layer to separate from the water phase for a minimum of 10
         min.  If the emulsion interface between layers is more  than one
         third the volume of the solvent layer, the analyst must employ
         mechanical techniques to complete the phase separation.  The
         optimum technique depends upon the sample, but may include
         stirring, filtration of the emulsion through glass wool, centri-
         fugation or other physical means.  Drain the aqueous phase into a
         1000-mL Erlenmeyer flask and collect the extract in a 250-mL
         ground-glass Erlenmeyer flask containing 2 ml of 37% potassium
         hydroxide solution.  Approximately 80 ml of the ethyl ether will
         remain dissolved in the aqueous phase.

   10.3  Add a 50-mL volume of ethyl ether to a sample bottle and repeat the
         extraction a second time, combining the extracts in the Erlenmeyer
         flask.  Perform a third extraction in the same manner.

   10.4  Add 15-mL reagent water and 1 or 2 clean boiling chips  to the
         250-mL flask and attach a three-ball Snyder column.  Prewet the
         Snyder column by adding 1 ml ethyl ether to the top. Place the
         apparatus on a hot water bath (60 to 65*C), such that the bottom of
         the flask is bathed in the water vapor.  Although the ethyl ether
         will evaporate in about 15 min, continue heating for a  total of 60
         min, beginning from the time the flask is placed on the water
         bath.  Remove the apparatus from the bath and let stand at room
         temperature for at least 10 min.

   10.5  Transfer the solution to a 60-mL separatory funnel  using 5 to 10 ml
         of reagent water.  Wash the basic solution twice by shaking for one
         min with 20-mL portions of ethyl ether.  Discard the organic
         phase.  The acid salts remain in the aqueous phase.

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   10.6  Acidify the contents of the separatory  funnel  to pH 2 by  adding  2
         fit of cold (4*C) sulfuric add  (1+3).   Test  the pH with Indicator
         paper.  Add 20 ml ethyl ether and  shake vigorously for 2  min.
         Drain the aqueous layer Into the 250-mL Erlenmeyer, then  pour the
         organic layer into a 125-mL Erlenmeyer  flask containing about 5  g
         of acidified, anhydrous sodium  sulfate.  Repeat the extraction
         twice more with 10-mL aliquots  of  ethyl  ether, combining  all
         solvent in the 125-mL flask.  Allow  the extract to remain in
         contact with the sodium sulfate for  approximately 2 h.

   10.7  Assemble a Kuderna-Oanish (K-0) concentrator by attaching a 10-mL
         concentrator tube to a 500-mL evaporative flask.

   10.8  Pour the combined extract through  a  funnel plugged with acid
         washed glass wool, and collect  the extract in  the K-0 concentra-
         tor.  Use a glass rod to crush  any caked sodium sulfate during the
         transfer.  Rinse the Erlenmeyer flask and column with 20  to 30 ml
         of ethyl ether to complete the  quantitative transfer.

   10.9  Add 1 to 2 clean boiling chips  to  the evaporative flask and attach
         a three-ball Snyder column.  Prewet  the  Snyder column by  adding
         about 1 ml ethyl ether to the top.   Place the  K-0 apparatus on a
         hot water bath, 60 to 65*C, so  that  the  concentrator tube is
         partially immersed in the hot water, and the entire lower rounded
         surface of the flask is bathed  with  hot  vapor.  Adjust the vertical
         position of the apparatus and the water  temperature as required  to
       .  complete the concentration in 15 to  20 min.  At the proper rate  of
         distillation the balls of the column will actively chatter but the
         chambers will not flood.  When  the apparent volume of liquid
         reaches 1 ml, remove the K-0 apparatus  and allow it to drain and
         cool for at least 10 min.

  10.10  Remove the Snyder column and rinse the  flask and its lower joint
         into the concentrator tube with 1 to 2 ml of ethyl  ether using a
         disposable pipet or syringe.  Add 1  ml MTBE and a fresh boiling
         chip.  Attach a micro-Snyder column  to  the concentrator tube and
         prewet the column by adding about 0.5 mL of ethyl  ether to the
         top.  Place the micro K-0 apparatus  on  the water bath so that the
         concentrator tube is partially  immersed  in the hot water.   Adjust
         the vertical position of the apparatus  and the water temperature as
         required to complete concentration in 5  to 10 min.   When the
         apparent volume of liquid reaches 0.5 ml, remove the micro K-0 from
         the bath and allow it to drain  and cool.  Remove the micro Snyder
         column and rinse the walls of the concentrator tube while adjusting
         the volume to 3-4 ml with MTBE.

11. ESTER IFICATION OF ACIDS

    11.1 Assemble the diazomethane generator  shown in Figure 1 in a hood.
         The collection vessel is a 10-15 ml  vial, equipped  with a
         teflon-lined screw cap and maintained at 0-5*C.

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    11.2 Add a sufficient amount of ethyl ether to tube 1 to cover the first
         Impinger.  Add 5 ml of MBTE to the collection vial.  Set the
         nitrogen flow at 5-10 cnn/mln.  Add 2 ml Dlazald solution (Sect.
         6.13) and 1.5 mL of 37% KOH solution to the second implnger.
         Connect the tubing as shown and allow the N£ flow to purge the
         dlazomethane from the reaction vessel Into the collection vial for
         30 m1n.  Cap the vial when^collection is complete and maintain at
         0-5*C.  When stored at 0-5*C this dlazomethane solution may be used
         over a period of 48 h.

    11.3 To each concentrator tube containing sample or standard, add 250 uL
         methanol and 0.5 ml of dlazomethane solution (Sect. 11.2).  Dilute
         to 5.0 ml with WTBE.

    11.4 Seal the concentrator tubes with teflon or glass stoppers and store
         in the hood at room temperature for 5 min.

    11.5 Open the concentrator tube and destroy any unreacted diazomethane
         by adding approximately 0.2 g activated silica to the samples.  The
         samples are now ready for analysis by GC.  Analyze as soon as
         possible.  The samples may be stored in the stoppered concentrator
         tubes at 0-4*C if the analysis cannot be performed immediately.
         The analysis should be performed within 24 hours.

12. GAS CHROMATOGRAPHY

    12.1 Tables 2 and 3 summarize the recommended operating conditions for
         packed and capillary chromatography.  Tables 1 and 2 contain
         estimated retention times and detection limits that can be achieved
       •  by this method.  Examples of the separations achieved for the
         methyl esters are shown in Figures 2 and 3.  Other columns,
         chromatographic conditions, or detectors may be used if the
         requirements of Section 8.3 are met.

    12.2 Calibrate the system daily as described in Section 7.

    12.3 Inject 1 to 5 wL of the sample extract for packed columns or 1-2 uL
         for capillary columns.  Record the resulting peak size in area or
         peak height units.  An automated system that consistently injects a
         constant volume of extract may also be used.

    12.4 The width of the retention time window used to make identifica-
         tions should be based upon measurements of actual retention time
         variations of standards over the course of a day.  Three times the
         standard deviation of a retention time can be used to calculate a
         suggested window size for a compound.  However, the experience of
         the analyst should weigh heavily in'the interpretation of
         chromatograms.

    12.5 If the response for the peak exceeds the working range of the
         system, dilute the extract and reanalyze.

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13. CALCULATIONS

    13.1  Determine the concentration of Individual compounds 1n the
          sample.  Calculate the amount of the free acid Injected from the •
          peak response using the calibration curve or calibration factor in
          Section 7.2.2.  The concentration in the sample can be calculated
          as follows:

                        Concentration, wg/L »    (A) (V )
                                                       t
                                                 (V ) (V J
                                                   1    s
         where:
            A a Amount of material injected, in nanograms.

            V^ 3 Volume of extract injected in vl.

            V^ » Volume of total extract in uL.

            V  - Volume of water extracted in liters.
    13.2 Report results in micrograms per liter as acid equivalent without
         further correction for recovery data.   When duplicate and spiked
         samples are analyzed, report all data  obtained with the sample
         results.

    13.3 For samples processed as part of a set where the laboratory spiked
         sample recovery falls outside of the control limits in Section 8.4,
         data for the affected parameters should be considered unacceptable.

14. METHOD PERFORMANCE

    14.1 The MDL is defined as the minimum concentration of a substance that
         can be measured and reported with 99%  confidence that the value is
         above zero.l®  The detection limits listed in Tables 1 and 2 were
         obtained from reagent water with an electron capture detector&

    14.2 In single laboratory evaluations, the  average recoveries presented
         in Tables 4 and 5 were obtained for Column 1 of Table 1 and column
         3 of Table 2.H  The standard deviations of the percent
         recoveries of these measurements are also included in Tables 4 and
         5.

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REFERENCES

1.  Goerlitz, D.G. and Lamar, W.L., "Oetennlnation of Phenoxy Acid
    Herbicides in Water by Electron-Capture and Microcoulometric Gas
    Chromatography," U. S. Geol. Survey Water Supply Paper 1817-C (1967).

2.  ASTM Annual Book of Standards, Part 31, 03694, "Standard Practice for
    Preparation of Sample Containers and for Preservation," American Society
    for Testing and Materials, Philadelphia, PA, p. 679, 1980.

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

4.  "OSHA Safety and Health Standards, General Industry," (29CFR1910),
    Occupational Safety and Health Administration, OSHA 2206, (Revised,
    January 1976).

5.  "Safety in Academic Chemistry Laboratories," American Chemical Society
    Publication, Committee on Chemical Safety, 3rd Edition, 1979.

6.  "Handbook for Analytical Quality Control in Water .and Wastewater
    Laboratories," EPA-600/4-79-019, U.S. Environmental Protection Agency,
    Environmental Monitoring and Support Laboratory - Cincinnati, Ohio
    45268, March 1979.

7.  ASTM Annual Book of Standards, Part 31, D3370, "Standard Practice for
    Sampling Water," American'Society for Testing and Materials,
    Philadelphia, PA, p. 76, 1980.

8.  McNair, H.M. and Bonelli, E.J., "Basic Chromatography," Consolidated
    Printing, Berkeley, California, p. 52, 1969.

9.  Eichelberger, O.W., Harris, L.E., Budde, W.L.  "Reference Compound to
    Calibrate Ion Abundance Measurement in Gas Chromatography-Mass
    Spectrometry," Analytical Chemistry. £7, 995 (1975).

10. Glaser, J.A., D.L. Foerst, G.D. McKee, S.A. Quave, and W.L. Budde,
    "Trace Analyses for Wastewaters," Environ. Sci. Techno!.. 15, 1426, 1981.

11. Pressley, T.A. and Longbottom, J.E., "The Determination of Chlorinated
    Herbicides in Industrial and Municipal Wastewater - Method 615."
    Environmental Monitoring and Support Laboratory, U. S. Environmental
    Protection Agency, Cincinnati, OH 45268, January 1982.

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                                    TABLE  1

                CHROMATOGfcAPHIC  CONDITIONS AND  DETECTION  LIMITS
                              CAPILLARY COLUMNS
  Analyte
(as methyl ester)

Dalapon
Dlcamba
2,4-D
PCP
2,4,5-TP
2,4.5-T
2,4-DB
Dinoseb
Picloram
  Retention Time, min.
Column 1*       Column 2*
 3.78
12.32
14.63
18.65
18.98
19.76
22.78
23.47
26.46
 2.99
10.89
13.78
16.98
18.65
20.25

24.61
32.33
Estimated Detection*
Limit (EDL) uq/L

        .001
        .01
        .01
        .0005
        .01
        .01
        .07
        .01
        .07
* The EDL is defined here as the student t factor times the standard
  deviation of at least 7 replicate analyses, of a 1 nt calibration standard
  at a concentration near but above the EDL.  These EDL values were obtained
  on Column 1.

* Column 1:  DB-1

  Column 2:  DB-5

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                                    TABLE 2

            CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
                                PACKED COLUMNS
Analyte
(as methyl ester)
Dicamba
2,4-0
2,4,5-TP
2,4,5-T
2,4-08
Dinoseb
Column 3
1.2
2.0
2.7
3.4
4.1
11.2
Retention Time
(min.)
Column 4
1.0
1.6
2.0
2.4
—
^ •
Method
Detection
Limit uq/L
0.27
1.2
0.17
0.20
0.91
0.07
Column 3 conditions:  Supelcoport (100/120 mesh) coated with 1.5% SP-2250/
1.95% SP-2401 packed in a 1.8 m long x 4 mm ID glass column with 95%
argon/5% methane carrier gas at^a flow rate of 70 mL/min.  Column
temperature:  isothermal at 185*C.  An electron capture detector was used to
measure MDL.

Column 4 conditions:  Gas Chrom Q (100/120 mesh) coated with 5% OV-210
packed in a 1.8 m long x 4 mm ID glass column with 95% argon/5% methane
carrier gas at a flow rate of 70 mL/min.  Column temperature:  isothermal at
185*C.

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                                    TABLE 3

                 Recommended Capillary GC Operating  Conditions

Column Type:                                  08-1 or OB-5
Film Thickness:                               0.25 »m
Column Dimension:                             30 m x 0.32 mm
Helium Linear Velocity:                       28-29 cm/sec
Injection Port Temp:                          200*C
Detector Temp:                                Column 3 - 290*C
                                              Column 4 - 200*C
Temperature Program:                          Column 1 - DB-1.  Inject at
                                              100*C and program immediately
                                              at 12"C/min to 200*C and hold
                                              until picloram elutes.
                                              Column 2 - DB-5.  Inject at
                                              80*C and hold 2 min.  Program
                                              at 16*C/min to 160*C and hold
                                              until picloram elutes.

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                                   TABLE 4

           SINGLE  OPERATOR ACCURACY AND PRECISION AS  DETERMINED BY
                            CAPILLARY COLUMN GCa
Analyte
Dalapon
Dicainba
2,4-0
PCP
Si 1 vex
2,4,5-T
2,4-DB
Dinoseb
Picloram
Spike Levelb
uq/L
4.05
4.13
4.01
1.01
4.05
4.23
4.02
4.12
4.01
AVGC
Recovery %
91
87
87
63
87
85
63
56
62
* RSO
9
11
11
11
13
13
32
19
14
aoata obtained on column 1

&The matrix is reagent water

CAT! results based on seven replicate analyses

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                                   TABLE 5.

                    SINGLE OPERATOR ACCURACY AND PRECISION*
                             FOR PACKED COLUMN GC
Parameter
2,4,0


2,4-DB


Dicamba


2,4,5-T


2,4,5-TP


Oalapon
-
Di noseb

Sample
Type
RW
DW
OW
RW
DW
DW
RW
DW
DW
RW
DW
DW
RW
DW
DW
RW
DW
DW
DW
Spike
- (uq/L)
10.9
10.1
200
10.3
10.4
208
1.2
1.1
22.2
1.1
1.3
25.5
1.0
1.3
25.0
23.4
23.4
0.5
102
Mean
Recovery
(*)
75
77
65
93
93
77
79
86
82
85
83
78
88
88
72
66
96
86
81
Standard
Deviation^)
4
4
5
3
3
6
7
9
6
6
4
5
5
4
5
8
13
4
3
*A11 results based upon seven replicate analyses.

RW » Reagent water
DW » Drinking water

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    nllrog«n
rubb«r  itopp*r
                     tub* I

 Figure 1.   Dlazomethane generator.
tub* 2
                                                                  glott tubing
                                     Collection
                                                                   Thermos or
                                                                   cryogenic cooler

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                 012345
                   Retention Time (M1n)

Figure 2.  Gas chroroatogram of methyl esters of chlorinated
           herbicides on Column 1.  For conditions, see Table 1

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                                         XI
VI
_*«
" •
61
                         o


                         M
       to
                                        "I
                                         °\
                                        «l
                              CM
                                                           2
                                                           o
                                                 CD
                                                 O
                                                 CM
§
                                                    •t>
                                                    •*
                                                     •
                                                    n
Figure 3.  Gas  Chromatogram of MethyT Esters  of Chlorinated Herbicides
           on CoTumrTB.  For Conditions, "See  Table 4.

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