4>EPA
     United States
     Environmental Protection
     Agency
Method 523: Determination of Triazine Pesticides and their
Degradates in Drinking Water by Gas Chromatography/Mass
Spectrometry (GC/MS)
Office of Water (MLK 140) EPA Document No. 815-R-l 1-002 February 2011 http://www.epa.gov/safewater/

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METHOD 523     DETERMINATION OF TRIAZINE PESTICIDES AND THEIR
                 DEGRADATES IN DRINKING WATER BY GAS
                 CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS)
                                  Version 1.0
                                 February 2011
M. M. Domino (Sullivan International Group, Inc.)
B. V. Pepich (U.S. EPA, Region 10 Laboratory)
D. J. Munch (U.S. EPA, Office of Ground Water and Drinking Water)
                         TECHNICAL SUPPORT CENTER
                 STANDARDS AND RISK MANAGEMENT DIVISION
               OFFICE OF GROUND WATER AND DRINKING WATER
                 U. S. ENVIRONMENTAL PROTECTION AGENCY
                            CINCINNATI, OHIO 45268
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                                      METHOD 523

      DETERMINATION OF TRIAZINE PESTICIDES AND THEIR DEGRADATES IN
  DRINKING WATER BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS)


1.   SCOPE AND APPLICATION

    1.1   This is a gas chromatography/mass spectrometry (GC/MS) method for the determination of
         triazine pesticides and their degradation products in finished drinking waters.  Precision and
         accuracy data have been generated for the method analytes in reagent water, drinking water
         from a groundwater source, and drinking water from a surface water source. The single
         laboratory Lowest Concentration Minimum Reporting Level (LCMRL) has also been
         determined in reagent water.  The following compounds can be determined using this
         method:

                                              Chemical Abstract Services
                         Analvte              Registry Number (CASRN)
                         Atrazine                        1912-24-9
                     Atrazine-desethyl                    6190-65-4
               Atrazine-desethyl-desisopropyl              3397-62-4
                  Atrazine-desisopropyl                  1007-28-9
                        Cyanazine                        21725-46-2
                        Propazine                         139-40-2
                        Simazine                         122-34-9
                  Terbuthylazine-desethyl                 30125-63-4
                      Terbuthylazine                     5915-41-3
                        Prometon                        1610-18-0
                        Prometryn                        7287-19-6
                        Ametryn                         834-12-8
                        Simetryn                        1014-70-6

    1.2   The MS conditions described in this method were developed using a time-of-flight (TOF)
         GC/MS system.  The method was validated at a second laboratory that used a quadrupole-
         based GC/MS system.

    1.3   The single laboratory LCMRL is the lowest spiking concentration such that the probability of
         spike recovery in the 50% to 150% range is at least 99%.  Single laboratory LCMRLs for the
         analytes in this method ranged from 0.40 to 2.1 micrograms per liter (ng/L), and are listed in
         Table 4 (all Tables are found in Section 17). The procedure used to determine the LCMRL is
         described elsewhere.1
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    1.4  Laboratories using this method are not required to determine an LCMRL, but they must
         demonstrate that the Minimum Reporting Level (MRL) for each analyte meets the
         requirements described in Section 9.2.4.

    1.5  Detection limit (DL) is defined as the statistically calculated minimum concentration that can
         be measured with 99% confidence that the reported value is greater than zero.2  The DL is
         dependent on sample matrix, fortification concentration, and instrument performance.
         Determining the DL for analytes in this method is optional (Sect. 9.2.6). DLs for method
         analytes fortified into RW ranged from 0.10 to 0.69 |ig/L. These values are presented in
         Table 4.

    1.6  This method is intended for use by analysts skilled in solid-phase extractions (SPE), the
         operation of GC/MS instrumentation, and the interpretation of the associated data.

    1.7  METHOD FLEXIBILITY - In recognition of technological advances in analytical
         instrumentation and techniques, the laboratory  is permitted to modify the GC and MS
         conditions  Changes may not be made to sample collection and preservation (Sect. 8), to
         the sample extraction procedure (Sect. 11.3), or to the quality control (QC)
         requirements (Sect. 9).  Method modifications must be considered only to improve method
         performance.  Modifications that are introduced in the interest of reducing cost or sample
         processing time, but result in poorer method performance, may not be used. In all cases
         where method modifications are proposed, the  analyst must perform the procedures outlined
         in the Initial Demonstration of Capability [(IDC), Sect. 9.2], verify that all QC acceptance
         criteria in this method (Tables 10 and 11) are met, and that method performance can be
         verified in a real sample matrix (Sect. 9.4).

2.  SUMMARY OF METHOD

    2.1  Samples  are pH adjusted and dechlorinated with ammonium acetate (MLX^HsC^) and pro-
         tected from microbial degradation using 2-chloroacetamide during sample collection.  Sam-
         ples are fortified with isotopically enriched surrogates [Atrazine-desethyl-desisopropyl(13C3),
         Atrazine-desisopropyl-t/5(ethyl-J5), Cyanazme-d5(N-ethy\-d5)., and Simazine-Jio(diethyl-Jio)]
         just prior to extraction. Analytes are extracted from a 250-milliliter (mL) sample aliquot
         using 250-milligram (mg) carbon cartridges. After extraction, the cartridges are dewatered
         with a small volume of methanol (MeOH) and  then eluted with 2 mL of ethyl acetate
         followed by two, 6-mL aliquots of 9:1 (v:v) dichloromethane/methanol (DCM/MeOH). The
         extracts are dried using anhydrous sodium sulfate (Na2SC>4) and concentrated using a stream
         of nitrogen gas.  Isotopically labeled internal standards (IS) [Atrazine-^ethyl-Js) and
         Atrazine-desethyl-Jy^sopropyl-Jy)] are added,  and the extracts brought to a final 1.0-mL
         volume.  Extracts are analyzed using a GC/MS operated in full scan mode. Method analytes
         are identified by comparing retention times and the acquired mass spectra to retention times
         and reference spectra for calibration standards acquired under identical  GC/MS conditions.
         The  concentration of each method analyte is determined using the IS technique.
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3.   DEFINITIONS

    3.1  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 (CCC) Standards. Additional CCCs may be required depending on the
         length of the Analysis Batch and/or the number of field samples.

    3.2  CALIBRATION STANDARD (CAL) - A solution of the method analytes prepared from the
         primary dilution standard(s) (PDS) and stock standard solution(s), which includes the ISs and
         SURs.  The CAL solutions are used to calibrate the instrument response with respect to
         analyte concentration.

    3.3  CONTINUING CALIBRATION CHECK (CCC) - A CAL containing the method analytes,
         surrogates, and ISs, which is analyzed periodically to verify the accuracy of the existing
         calibration.

    3.4  DETECTION LIMIT (DL) - The minimum concentration of an analyte that can be
         identified, measured  and reported with 99% confidence that the analyte concentration is
         greater than zero.  This is a statistical determination (Sect. 9.2.6), and accurate quantitation is
         not expected at this level.

    3.5  EXTRACTION BATCH - A set of up to 20 field samples (not including QC samples)
         extracted together by the same person(s) during a work day using the same lot of SPE
         devices, solvents, surrogate solution, and fortifying solutions.  Required QC samples include
         Laboratory Reagent Blank,  Laboratory Fortified Blank, Laboratory Fortified Matrix, and
         either a Field Duplicate or Laboratory Fortified Sample Matrix Duplicate.

    3.6  FIELD DUPLICATES (FD1 and FD2) - Separate samples collected at the same time, and
         shipped and stored under identical conditions. Method precision, including the contribution
         from sample collection procedures, is estimated from the analysis of FDs.  For the purposes
         of this method, Field Duplicates are necessary to conduct repeat analyses if the original field
         sample is lost, or to conduct repeat analyses in the case of QC failures associated with the
         analysis of the original field sample.  Field Duplicates are used to prepare Laboratory
         Fortified Sample Matrix (Sect. 3.9) and Laboratory Fortified Sample Matrix Duplicate (Sect.
         3.10) QC samples.

    3.7  INTERNAL STANDARD (IS) - A pure compound added to all standard solutions, field
         samples and QC samples in a known amount. Each internal standard is assigned to a specific
         analyte or multiple analytes, and is used to measure relative response.

    3.8  LABORATORY FORTIFIED BLANK (LFB) - An aliquot of reagent water to which known
         quantities of the method analytes are added.  The LFB is extracted and  analyzed as a sample
         including use of the preservation procedures in Section 8. The LFB is used during the IDC to
         verify method performance for precision and accuracy, and as an ongoing QC element.
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3.9   LABORATORY FORTIFIED SAMPLE MATRIX (LFSM) - A Field Duplicate to which
     known quantities of the method analytes are added. The LFSM is extracted and analyzed as a
     sample, and its purpose is to determine whether the sample matrix contributes bias to the
     analytical results.

3.10 LABORATORY FORTIFIED SAMPLE MATRIX DUPLICATE (LFSMD) - A second
     Field Duplicate of the same field sample used to prepare the LFSM which is fortified
     identically to the LFSM. The LFSMD is used instead of the FD to assess method precision
     and accuracy when the occurrence of the method analytes at a concentration greater than the
     MRL is infrequent.

3.11 LABORATORY REAGENT BLANK (LRB) - An aliquot of RW that is treated exactly as a
     sample including exposure to all glassware, equipment, solvents, reagents, sample
     preservatives, ISs, and SURs associated with the Extraction Batch. The LRB is used to
     determine if method analytes or other interferences are introduced via the laboratory
     environment, the reagents, or the apparatus.

3.12 LOWEST CONCENTRATION MINIMUM REPORTING LEVEL (LCMRL) - The single-
     laboratory LCMRL is the lowest spiking concentration such that the probability of spike
     recovery in the 50 and 150% range is at least 99% l

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

3.14 MINIMUM REPORTING LEVEL (MRL) - The minimum concentration that can be
     reported by  a laboratory as a quantified value for the method analyte in a sample following
     analysis. This concentration must meet the criteria defined in Section 9.2.4 and must be no
     lower than the concentration of the lowest CAL for the method analyte. A laboratory may be
     required to demonstrate a specific MRL by a regulatory body if this method is being
     performed for compliance purposes.

3.15 PRIMARY DILUTION STANDARD (PDS) - A solution containing the method analytes (or
     internal standards or surrogate analytes) prepared in the laboratory from Stock Standard
     Solutions and diluted as needed to prepare calibration standards and sample fortification
     solutions.

3.16 QUALITY CONTROL SAMPLE (QCS) - A solution containing the method analytes at a
     known concentration, which is obtained from a source external to the laboratory and different
     from the source of calibration standards. The purpose of the QCS is to verify the accuracy of
     the primary  calibration standards.

3.17 REAGENT WATER (RW) - Purified water that does not contain any measurable quantity of
     the method analytes or interfering compounds at or above  1/3 the MRL.
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    3.18 STOCK STANDARD SOLUTION - A concentrated solution containing one or more of the
         method analytes that is prepared in the laboratory using assayed reference materials or
         purchased from a reputable commercial source, so that the concentration and purity of
         analytes are traceable to certificates of analysis.

    3.19 SURROGATE ANALYTE (SUR) - A pure analyte, extremely unlikely to be found in any
         sample, that is added to a sample aliquot in a known amount before extraction, and which 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.

4.   INTERFERENCES

    4.1  All glassware must be meticulously cleaned. Wash glassware with detergent and tap water,
         rinse with tap water, followed by RW. Volumetric glassware must be solvent rinsed after
         washing and dried in a low temperature oven [<120 degrees Centigrade (°C)] or air-dried.
         Non-volumetric glassware may be heated in a muffle furnace at 400 °C for two hours  as a
         substitute for a solvent rinse.

    4.2  Method interferences may  be caused by contaminants in solvents, reagents (including RW),
         sample bottles and caps, and other sample processing hardware.  These interferences may
         lead to discrete artifacts and/or elevated baselines in the chromatograms. All laboratory
         reagents and equipment must be routinely demonstrated to be free from interferences (less
         than 1/3 the MRL for the method analytes) under the conditions of the analysis. This may be
         accomplished by analyzing LRBs as described in Section 9.3.1.

    4.3  Preservatives (Sect.  8.1) are added to samples to ensure the stability of method analytes
         during shipping and storage prior to analysis.  The potential exists for trace-level organic
         contaminants in these reagents.  Interferences from these sources must be evaluated by
         analysis of LRBs, particularly when new lots of reagents are acquired.

    4.4  SPE cartridges may be a source of interferences. The analysis of LRBs can provide
         important information regarding the presence or absence of such interferences.  Brands and
         lots of SPE devices must be tested to ensure that contamination does not preclude analyte
         identification and quantitation.

    4.5  Silicone compounds may be leached from punctured septa of autosampler vials.  This can
         occur after repeated injections from the same autosampler vial.  These silicone compounds,
         which appear as regularly spaced chromatographic peaks with similar fragmentation patterns,
         could unnecessarily  complicate the total ion chromatograms and may interfere with the
         identification of the method analytes.

    4.6  Matrix interferences are caused by contaminants that are present in the sample.  The extent of
         matrix interferences will vary considerably from source to source depending upon the nature
         of the water. The analysis of Laboratory Fortified Sample Matrix (Sect.  9.3.7) provides
         evidence for the presence (or absence) of matrix effects.
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    4.7  This method uses a number of isotopically labeled ISs and SUR analytes.  These standards
         must be determined to be sufficiently free of the unlabeled parent molecule to permit accurate
         quantitation of field samples. This is verified during the IDC via analysis of LRBs.

5.   SAFETY

    5.1  The toxicity or carcinogenicity of each reagent used in this method has not been precisely
         defined. Each chemical must be treated as a potential health hazard and exposure to these
         chemicals should be minimized.  Each laboratory is responsible for maintaining an awareness
         of OSHA regulations regarding safe handling of chemicals used in this method.3'4 The
         OSHA laboratory standards can be found on line at
         http://www.osha.gov/SLTC/laboratories/standards.html. A reference file of MSDSs should
         be made available to all personnel involved in the chemical analysis.

    5.2  Pure standard materials and stock standard solutions of the method compounds should be
         handled with suitable protection for skin, eyes, etc.5

6.   EQUIPMENT AND SUPPLIES

    References to specific brands or catalog numbers are included as examples only and do not imply
    endorsement of the product. These references do not preclude the use of other vendors or supplies.

    6.1  SAMPLE CONTAINERS - Clean, amber bottles (250 mL or larger) fitted with polytetra-
         fluoroethylene (PTFE)-faced silicone septa and polypropylene  screw caps (I-Chem Cat.
         No. S249-0250 or equivalent).

    6.2  VIALS - Amber, 2-mL glass autosampler vials with PTFE-faced septa (Fisher Cat. No. 03-
         375-19B or equivalent).

    6.3  MICRO SYRINGES - Suggested sizes include 10, 25, 50, 100, 250, and 500 |iL.

    6.4  VOLUMETRIC FLASKS - Class A, suggested sizes include 5, 10, 50, 100, 200, and 500
         mL for preparation of reagents and standards.

    6.5  VOLUMETRIC PIPETTES - Class A, suggested sizes include 2, 3, 4, 6, and  10 mL.

    6.6  ANALYTICAL BALANCE - Capable of weighing to the nearest 0.0001 gram.

    6.7  SPE VACUUM MANIFOLD AND SAMPLE TRANSFER LINES - Supelco Visi-Prep™
         Part No. 57044 or equivalent, and Supelco Visi-Prep™ Part No. 57275 or equivalent.

    6.8  SPE CARTRIDGES - Cartridges packed with 250 mg of graphitized, non-porous carbon
         (Supelco ENVI-Carb™, Cat. No. 57092 or equivalent).

    6.9  AUTOMATED EXTRACTORS - An automated or robotic system designed for use with
         SPE cartridges may be used if all quality control requirements  discussed in Section 9 are met.

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     Automated systems may use either vacuum or positive pressure to process samples and
     solvents through the cartridges. All extraction and elution steps must be the same as in the
     manual procedure. Extraction and/or elution steps may not be changed or omitted to accom-
     modate the use of an automated system.

6.10 DISPOSABLE PASTEUR PIPETTES - Nine-inch borosilicate glass (Fisher Cat. No. 13-
     678-20C or equivalent).
6.11  DRYING COLUMN - The drying column must be able to contain 3 g of anhydrous
     plus several milliliters of eluate. The drying column must not leach interfering compounds or
     irreversibly adsorb method analytes.  Any small glass column may be used, such as a glass
     pipette with glass wool plug (Chase Scientific Glass, Inc., P1005, 4.5-mL Monstr-Pette,
     Fisher Part No. 22-378-893 or equivalent).

6.12  EXTRACT CONCENTRATION SYSTEM - Extracts are concentrated by blowdown with
     nitrogen gas using a water bath set at 35° to 40 °C (Meyer N-Evap, Model 111,
     Organomation Associates, Inc., or equivalent).  Other automated concentration devices may
     be used.

6.13  LABORATORY OR ASPIRATOR VACUUM SYSTEM - Sufficient capacity to maintain a
     vacuum of approximately 15 to 25 inches of mercury.

6.14  GRADUATED CONICAL SCREW-CAP TEST TUBES - Fifteen-mL capacity (Corning
     Part No. 8082-15, Fisher Part No. 05-538-30A, or equivalent), used to collect the eluate from
     the carbon cartridges.  Also, 40-mL capacity (Kimble Part No. 45200-40, Fisher Part No. 05-
     538-33B or equivalent), used to collect eluates from the drying columns.

6.15  GAS CHROMATOGRAPHY MASS SPECTROMETRY SYSTEM (GC/MS)

     6.15.1  FUSED SILICA CAPILLARY GC COLUMN - Fused silica capillary column
            [20-meter x 0. 18-millimeter (mm) inside diameter (i.d.)] coated with a 0.20-|im
            bonded film of poly methylphenyl siloxane (Restek Rtx®-50 or equivalent). Any
            column that provides adequate resolution, peak shape, capacity (Sect.  10.2.2.1 and
            10.2.2.2), and accuracy and precision (Sect. 9) may be used.  A mid-polarity, low-
            bleed column is recommended for use with this method to provide appropriate
            selectivity, and to minimize mass spectrometric background.

     6. 15.2  GC INJECTOR AND OVEN - Equipped for split/splitless injection (Agilent 6890
            GC with Agilent 7683 autosampler or equivalent).  Some of the analytes included in
            this method are very polar and/or subject to thermal breakdown in the injection port.
            This effect increases when the injector is not properly deactivated or operated at
            excessive temperatures. The injection system must not allow analytes to contact hot
            stainless steel or other active surfaces that promote decomposition. The performance
            data in Section 17 were obtained using hot, splitless injection with a 2-mm-i.d.
            quartz liner (Restek Cat. No. 20914).  Other injection techniques such as temperature
            programmed injections, cold on-column injections and large volume injections may

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                be used if the QC criteria in Section 9 are met. Commercially available inlet
                systems, specifically designed for these alternate types of injections, must be used if
                these options are selected.  The GC system must provide consistent sample injection
                volumes and be capable of performing temperature programming at a constant flow
                rate, constant linear velocity, or constant pressure.

                6.15.2.1  GC SYRINGE - During method development, a 5.0-|iL syringe was used
                         to inject 0.5-|iL aliquots of standards and extracts (Agilent Cat. No. 5181-
                         1273 or equivalent).

         6.15.3  GC/MS INTERFACE - Interface must allow the capillary column  or transfer line
                exit to be placed within a few millimeters of the ion source.

         6.15.4  MASS SPECTROMETER (MS) - The MS must be capable of electron ionization
                and collecting spectra in positive ion mode. The instrument must be capable of
                obtaining at least five scans during the chromatographic peaks. Ten to fifteen scans
                across chromatographic peaks are recommended.  The spectrometer must produce a
                mass spectrum that meets all criteria in Table 1 when a solution containing approxi-
                mately five nanograms (ng) of decafluorotriphenyl phosphine (DFTPP) is injected
                into the GC/MS.

         6.15.5  DATA SYSTEM - An interfaced data system is required to acquire, store, and
                output MS data.  The computer software must have the capability of processing
                stored GC/MS data by recognizing a chromatographic peak within  a given retention
                time window.  The software must allow integration of the ion abundance of any
                specific ion between specified time or scan number limits. The software must also
                allow construction of linear or second-order regression calibration  curves, and
                calculation of concentrations using the internal standard technique.

7.   REAGENTS AND STANDARDS

    7.1  REAGENTS AND SOLVENTS - Reagent-grade or better chemicals must  be used. Unless
         otherwise indicated,  it is intended that all reagents will conform to the specifications of the
         Committee  on Analytical Reagents  of the American Chemical Society (ACS), where such
         specifications are available.  Other grades may be used, as long as the reagent is of
         sufficiently high purity to permit its use without negatively affecting data quality.

         7.1.1   HELIUM -  99.999% or better, GC carrier gas.

         7.1.2   REAGENT  WATER - Purified water, which does not contain any  measurable
                quantities of any method analytes or interfering compounds  at or above 1/3 the MRL
                for each  compound of interest.

         7.1.3   METHANOL - (MeOH) (CASRN 67-56-1) - High purity, demonstrated to be free
                of analytes and interferences (Fisher GC Resolv®-grade or equivalent).
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     7.1.4   ETHYL ACETATE (EtOAc) (CASRN 141-78-6) - High purity, demonstrated to be
            free of analytes and interferences (B & J Brand GC2®, Capillary GC/GC-MS-grade
            or equivalent).

     7.1.5   DICHLOROMETHANE (DCM), also known as METHYLENE CHLORIDE
            (MeCl2) (CASRN 75-09-2) - High purity, demonstrated to be free of analytes and
            interferences (B & J Brand GC2®, Capillary GC/GC-MS-grade or equivalent).

     7.1.6   SODIUM SULFATE, ANHYDROUS (CASRN 7757-82-6) - Soxhlet extracted with
            DCM for a minimum of four hours or heated to 400 °C for two hours in a muffle
            furnace. An "ACS grade, suitable for pesticide residue analysis," or equivalent, of
            anhydrous Na2SO4 is recommended (Fisher, Cat. No. S415-1 OS or equivalent).

     7.1.7   DICHLOROMETHANE/METHANOL 9:1 - Elution solvent.  To make 1 L, add
            900 mL of DCM to 100 mL of MeOH and mix thoroughly. Store in an amber glass
            container.

     7.1.8   DICHLOROMETHANE/ETHYL ACETATE 3:1- Drying column rinse solvent.
            To make 1 L, add 750 mL of DCM to 250 mL of EtOAc and mix thoroughly. Store
            in an amber glass container.

     7.1.9   AMMONIUM ACETATE (CASRN 631 -61 -8) - High purity, demonstrated to be
            free of analytes and interferences (Sigma Aldrich, Cat. No. A7262 or equivalent).

     7.1.10  2.5 M AMMONIUM ACETATE CONCENTRATED STOCK SOLUTION
            (192 g/L) - Used to sequester free available chlorine  and to buffer field samples.  To
            prepare 200 mL of solution, add 38.5 g ammonium acetate to a 200-mL volumetric
            flask, then add RW to the mark and mix well.

     7.1.11  2-CHLOROACETAMIDE (C1CH2CONH2) (CASRN 79-07-2) - Used as an anti-
            microbial agent. High purity, demonstrated to be free of analytes and interferences
            (Sigma Cat. No. C0267 or equivalent).

7.2   STANDARD SOLUTIONS - When a compound's purity is assayed to be 96% or 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 only as examples. Stock standard solutions are estimated to be stable for at least
     six months if stored at -10 °C or colder. Any fortified or dilute solutions made from the stock
     standards are stable for at least 60 days provided they are stored at a temperature <-10 °C and
     the stock standard solutions have not exceeded their six month stability period. Although
     estimated stability times for standard solutions are given, laboratories should use
     accepted  QC practices to determine when their standards  need to be replaced.

     7.2.1   INTERNAL STANDARD SOLUTIONS - This method uses isotopically enriched
            ISs, specifically Atrazine-J5(ethyl-J5) (CASRN 163165-75-1; CDN Isotopes Cat.
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No. D4389 or equivalent) and Atrazine-desethyl-J7(isopropyl-J7) (CASRN 6190-65-
4; CDN Isotopes Cat. No. D5639  or equivalent).

7.2.1.1  INTERNAL STANDARD STOCK SOLUTIONS (1000 ng/mL) - Prepare
        the stock standards individually by weighing 10 mg of the solid materials
        (Atrazine-t/s and Atrazine-desethyl-Jy) into tared 10-mL volumetric flasks
        and dilute to volume with EtOAc.  Sonication may be required to achieve
        full dissolution of the solid materials. Alternatively, commercially
        produced standard solutions may be used.

7.2.1.2  INTERNAL STANDARD PRIMARY DILUTION STANDARD (IS PDS)
        (200 ng/mL) - Prepare the IS PDS by adding enough of each internal
        standard stock solution to a volumetric flask partially filled with EtOAc to
        make the final concentrations 200 |j,g/mL when filled to the mark with
        EtOAc. During method development, addition of 10 microliters (|jL) of the
        IS PDS to each 1.0-mL extract produced a final concentration of 2.0
        |ig/mL. Analysts are permitted to use other PDS concentrations and
        volumes provided all field samples and calibration standards contain the
        same amount of IS, the concentration of the IS added provides adequate
        signal to maintain precision, and the volume added has a negligible effect
        on the final concentration. Analysts are NOT permitted to use alternate
        internal standards.

SURROGATE STOCK STANDARD SOLUTION (1500, 500, or 100 ng/mL) -
This method uses isotopically enriched compounds for surrogate standards,
specifically Atrazine-desethyl-desisopropyl(13C3) (Cambridge Isotope Labs, Cat. No.
CLM-7528-0), Atrazine-desisopropyl-J5(ethyl-J5) (CDN Isotopes Cat. No. D6456),
Cyanazine-J5(jV-ethyl-J5) (CASRN 3397-62-4; CDN Isotopes Cat. No.  D6136), and
Simazine-dio(diethyl-dio) (CASRN 220621-39-6; CDN Isotopes Cat. No. D5654).
Concentrations were chosen based on the solubility of these analytes in EtOAc, on
their relative instrument response, and in the case of Atrazine-desethyl-desisopropyl
(13C3), on other important chromatographic considerations (Sect. 10.2.2.2). Prepare
single component SUR stock standards by weighing out approximately 15 mg
Cyanazine-t/s, and 5.0 mg of Atrazine-desisopropyl-t/s and Simazine-t/io solid
materials using an analytical balance into separate, tared 10-mL volumetric flasks.
Dilute to volume with EtOAc.  Atrazine-desethyl-desisopropyl (13C3) surrogate is
much less  soluble in EtOAc and is prepared by weighing out approximately 5.0 mg
of the solid using an analytical balance into a tared 50-mL volumetric flask and
diluting to volume with EtOAc; the Atrazine-desethyl-desisopropyl (13C3) used
during method development was custom synthesized by Cambridge Laboratories.
Analysts are NOT permitted to use alternate surrogates.

7.2.2.1  SURROGATE PRIMARY DILUTION STANDARDS (SUR PDS) -
        Prepare a single SUR PDS that contains Simazine-t/io and Atrazine-
        desisopropyl-t/s at 200 ng/mL, and Cyanazine-t/s at 500 |j,g/mL by making
        appropriate dilutions of the SUR stock standard solutions into ethyl acetate.

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                          During method development, the Atrazine-desethyl-desisopropyl (13C3)
                          SUR stock was used without further dilution to fortify samples prior to
                          extraction (Sect.  11.2.2) and to prepare calibration standards (Sect. 7.2.4).

          7.2.3    ANALYTE STANDARD SOLUTIONS - Obtain the analytes listed in the table in
                  Section 1.1 as neat standards. Prepare the analyte stock and Primary Dilution
                  Standards as described below.  Alternatively, commercially produced standard
                  solutions may be used.

                  7.2.3.1  ANALYTE STOCK STANDARD - Prepare the stock standards
                          individually by weighing  10-20 mg of the solid materials using an
                          analytical balance into tared 5-mL, 10-mL, or larger volumetric flasks and
                          diluting to volume with EtOAc.  Atrazine-desethyl-desisopropyl exhibits
                          limited solubility in EtOAc and must be prepared at a concentration of
                          <100 |j,g/mL. The table below summarizes the concentration of the analyte
                          stock standards used during method development. In some cases,
                          sonication is required to achieve full dissolution of the neat materials.
Analyte
Atrazine
Atrazine-desethyl
Atrazine-desisopropyl
Atrazine-desethyl-desisopropyl
Cyanazine
Propazine
Simazine
Terbuthylazine
Terbuthylazine-desethyl
Prometon
Prometryn
Ametryn
Simetryn
Analyte Stock Standard*
(Hg/mL)
2000
1000
500
100
2000
2000
500
2000
850
1200
900
1200
840
Manufacturer, Part No.**
Chem Service, PS-380
Chem Service, MET-380B
Chem Service, MET-58A
Chem Service, MET-58C
Chem Service, PS-387
Chem Service, PS-385
Chem Service, PS-58
Chem Service, PS-413
Chem Service, MET-413A
Chem Service, PS-386
Chem Service, PS-384
Chem Service, PS-383
Chem Service, PS-3 81
* Analyte stock standard concentrations used during method development. Some analytes are near their solubility
limits at the storage temperature and required sonication to dissolve.
** Other manufacturer's standards are allowed as long as they have acceptable purity.
                  7.2.3.2  ANALYTE PRIMARY DILUTION STANDARD (50 ng/mL) - The ana-
                          lyte PDS is used to prepare the CALs and to fortify the LFBs, LFSMs and
                          LFSMDs with the method analytes.  The analyte PDS is prepared by adding
                          appropriate volumes of the of the analyte stock solutions (except Atrazine-
                          desethyl-desisopropyl) into a single volumetric flask and diluting to volume
                          such that the final concentration is 50 |j,g/mL. Because the concentration of
                          Atrazine-desethyl-desisopropyl in the analyte stock is low due to its limited
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                         solubility in EtOAc, use the analyte stock directly as an analyte PDS
                         solution in the steps that follow.

         7.2.4   CALIBRATION STANDARDS - Prepare a calibration curve of at least five levels
                from dilutions of the analyte PDS in EtOAc.  The preparation scheme, with
                concentrations of CALs that were used to collect data in Section 17, is presented in
                the table below. A constant concentration of each IS and SUR analyte (in the range
                of 1 to 5 |ig/mL) is added to each calibration  solution.  For example, add 20 jiL of
                the IS PDS (Sect. 7.2.1.2) and 20 |iL of the multi-component SUR PDS (Sect.
                7.2.2.1) to each CAL.  For the surrogate Atrazine-desethyl-desisopropyl (13C3), add
                20 |jL of the SUR. stock standard (Sect. 7.2.2). The lowest concentration CAL must
                be at or below the MRL. The CAL standards may also be used as CCCs.  The
                standards must be stored at -10 °C or lower.
CAL
Level
1
2
3
4
5
6
7
8
Analyte PDS*
(ug/mL)
50
50
50
50
50
50
50
50
Analyte PDS Volume**
(uL)
4
8
12
20
30
40
80
200
Final CAL
Volume (mL)
2.00
2.00
2.00
2.00
2.00
2.00
2.00
2.00
CAL
(us/mL)
0.10
0.20
0.30
0.50
0.75
1.00
2.00
5.00
* For atrazine-desethyl-desisopropyl, the analyte stock standard was used at a concentration of 100 (ig/mL.
** For atrazine-desethyl-desisopropyl, 1A of the volume reported in this table was used.

         7.2.5   GC/MS TUNE CHECK SOLUTION (5 jig/mL) - Prepare a DFTPP (CASRN 5074-
                71-5) solution in DCM.  DFTPP is more stable in DCM than in acetone or EtOAc.
                Store this solution in an amber glass screw cap vial at -10 °C or lower.

8.   SAMPLE COLLECTION, PRESERVATION, AND STORAGE

    8.1   SAMPLE COLLECTION

         8.1.1   Prior to shipment to the field, the ammonium acetate solution and 2-chloroacetamide
                must be added to each sample bottle.  250-mL sample bottles are recommended. For
                this sample volume, add 2.0 mL of the ammonium acetate concentrated stock (Sect.
                7.1.10) and 500 mg of 2-chloroacetamide. These reagents may be added in the field.
                However, the preservatives must be added to the container prior to sample collection.
                If other collection volumes are used, adjust the amount of the preservatives so that
                the final concentrations of ammonium acetate and 2-chloroacetamide in the sample
                containers are 1.5 g/L (20 millimolar, mM) and 2.0 g/L, respectively. Cap the bottles
                to avoid loss of the preservation reagents.
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                NOTE: The extraction procedure was developed using a 250-mL sample volume.
                Although the method performed well at this volume in matrices containing high
                levels of total organic carbon (TOC), larger extraction volumes could conceivably
                cause breakthrough for field samples with high TOC levels. For this reason, sample
                size must not exceed the recommended volume by more than 10%.  Smaller sample
                volumes, although not subject to breakthrough, may decrease method sensitivity
                making it more difficult to confirm the MRL (Sect. 9.2.4).

         8.1.2   When sampling from a cold water tap, open the tap and allow the system to flush
                until the water temperature has stabilized (usually 3  to 5 minutes). Collect a repre-
                sentative sample from the flowing system using a beaker of appropriate size.  Use
                this bulk sample to generate individual samples as needed.  Transfer a volume of at
                least 245 mL into each collection container, cap the  container, and invert it several
                times to mix the  sample with  the preservatives. Care must be taken not to overfill
                the bottle and flush out the preservation reagents.  Samples do not need to be head-
                space free.

         8.1.3   When sampling from an open body of water, fill a beaker with water sampled from a
                representative area.  Use this  bulk sample to generate individual samples as needed.

    8.2  FIELD DUPLICATES - Collect enough Field Duplicates  to fulfill QC requirements for
         LFSMs and LFSMDs (at least three identical samples).

    8.3  SAMPLE SHIPMENT AND STORAGE - Samples must be chilled during shipment and
         must not exceed 10 °C  during the first 48 hours after collection. Samples must be confirmed
         to be at or below  10 °C when they are received at the laboratory.  In the laboratory, samples
         must be stored at or below 6  °C until extraction.  Samples  must not be frozen.

    8.4  SAMPLE HOLDING TIMES - Samples should be analyzed as soon as possible. Samples
         that are collected and stored as described in Sections 8.1 and 8.3 may be stored prior to
         analysis for a maximum of 28 days. Extracts may be held for a maximum of 28 days prior to
         analysis, if they are stored at -10 °C or lower.

9.   QUALITY CONTROL

    9.1  QC requirements include the IDC and ongoing QC requirements. This section describes each
         QC parameter, its required frequency, and the performance criteria that must be met in order
         to meet EPA quality objectives. The QC criteria discussed in the following sections are
         summarized in Section 17, Tables 10  and 11.  These QC requirements are considered the
         minimum acceptable QC criteria. Laboratories are encouraged to institute additional QC
         practices to meet their specific needs.

    9.2  INITIAL DEMONSTRATION OF CAPABILITY - The IDC must be successfully
         performed prior to analyzing any field samples. Prior to conducting the IDC, the analyst
         must meet the calibration requirements outlined in Section 10. Requirements for the IDC are
         described in the following sections and are summarized in Table 10.

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9.2.1   DEMONSTRATION OF LOW SYSTEM BACKGROUND - Analyze an LRB.
       Confirm that the blank is free of contamination as defined in Section 9.3.1.

9.2.2   DEMONSTRATION OF PRECISION - Prepare, extract and analyze four to seven
       replicate LFBs. Fortify these samples near the midrange of the initial calibration
       curve. Ammonium acetate and 2-chloroacetamide must be added to the samples as
       described in Section 8.1. The percent relative standard deviation (RSD) of the
       concentrations of the replicate analyses must be <20% for all analytes.

              n, _ „_  Standard Deviation of Measured Concentrations  ,
              %RSD =	x 100
                                 Average Concentration

9.2.3   DEMONSTRATION OF ACCURACY - Calculate the average percent recovery
       using the same set of replicate data generated for Section 9.2.2. The average
       recovery of the replicate analyses must be within +30% of the true value.

                  .. _.          Average Measured Concentration  , ^
                  % Recovery =	x 100
                                   Fortified Concentration

9.2.4   MINIMUM REPORTING LEVEL (MRL) CONFIRMATION - Establish a target
       concentration for the MRL based on the intended use of the method. Analyze an
       initial calibration following the procedures in Section 10.2. The lowest CAL used to
       establish the initial calibration (as well as the low-level CCC) must be at or below
       the concentration of the MRL. Establishing the MRL concentration too low may
       cause repeated failure of ongoing QC requirements.  Confirm the MRL following  the
       procedure outlined below.

       9.2.4.1  Fortify, extract and analyze seven replicate LFBs at or below the target
               MRL concentration.  Ammonium acetate and 2-chloroacetamide must be
               added to the samples as described in Section 8.1.  Calculate the mean
               (Mean) and standard deviation (S) for these replicates. Determine the half
               range for the prediction interval of results (HRPIR) using the equation

                              HRPIR = 3.9638

               where S is the standard deviation, and 3.963 is a constant value for seven
               replicates.1

       9.2.4.2  Confirm that the upper and lower limits for the PIR (PIR = Mean ± HRPIR)
               meet the upper and lower recovery limits as shown below.

               The Upper PIR Limit must be <150% recovery.
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                              Mean + HRprrf
                                               xlOO<150%
                          Fortified Concentration

                The Lower PIR Limit must be >50% recovery.
                               Mean - HRprrf
                                          PIR     xlOO>50%
                           Fortified Concentration

       9.2.4.3   The MRL is validated if both the Upper and Lower PIR Limits meet the
                criteria described above.  If these criteria are not met, the MRL has been set
                too low and must be confirmed again at a higher concentration.

       NOTE: These equations are only valid for seven replicate samples.

9.2.5   QUALITY CONTROL SAMPLE - Analyze a mid-level QCS (Sect. 9.3.9) to
       confirm the accuracy of the primary CALs.

9.2.6   DETECTION LIMIT DETERMINATION (optional) - While DL determination is
       not a specific requirement of this method, it may be required by various regulatory
       bodies associated with compliance monitoring. It is the responsibility of the labora-
       tory to ascertain whether DL determination is required based upon the intended use
       of the data.

       Analyses for this procedure must be done over at least three days (both the sample
       extraction and the GC analyses must be done over a period of at least three days).
       Prepare at least seven replicate LFBs.  Use the solutions described in Section 7.2 to
       fortify at a concentration estimated to be near the DL. This concentration  may be
       estimated by selecting a concentration at two to five times the noise level.
       Ammonium acetate and 2-chloroacetamide must be added to the samples as
       described in Section 8.1. Process the seven replicates through all steps in  Section  11.

       NOTE:  If an MRL confirmation data set meets these requirements, DLs may be
       calculated from the MRL confirmation data, and no additional analyses are
       necessary.

       Calculate the DL using the following equation:

                                 DL= St(n.iji.a = 0.99)

       where:
       t(n-i,i-a = 0.99)=  Student's t value for the 99% confidence level with n-1 degrees of
                      freedom
       n = number of replicates
       S = standard deviation of replicate  analyses.


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            NOTE: Do not subtract blank values when performing DL calculations.

9.3   ONGOING QC REQUIREMENTS - This section describes the ongoing QC procedures that
     must be followed when processing and analyzing field samples. Table 11 summarizes these
     requirements.

     9.3.1   LABORATORY REAGENT BLANK - An LRB is required with each Extraction
            Batch. If within the retention time window of any analyte, the LRB produces a peak
            that would prevent the determination of that analyte, determine the source of
            contamination and eliminate the interference before processing samples.
            Background from analytes or contaminants that interfere with the measurement of
            method analytes must be less than 1/3 the MRL. If the method 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 that yielded a positive
            result.

            NOTE: Subtracting blank values from sample results is not permitted.

            NOTE: Although quantitative  data below the MRL may not be accurate  enough for
            data reporting, such  data are useful in determining the magnitude of any background
            interference. Therefore, blank contamination levels may be estimated by  extrapola-
            tion, when the concentration is below the MRL.

     9.3.2   CONTINUING CALIBRATION CHECK  - Analyze CCC standards at the
            beginning of each Analysis Batch, after every ten field samples, and at the end of the
            Analysis Batch. See Section  10.3 for concentration requirements and acceptance
            criteria.

     9.3.3   LABORATORY FORTIFIED BLANK - An LFB is required with each Extraction
            Batch. The fortified concentration of the LFB must be rotated between low,
            medium, and high concentrations from batch to batch. The low concentration LFB
            must be at or below  the MRL. Similarly, the high concentration LFB must be near
            the high end of the calibration range established during the initial calibration (Sect.
            10.2).  Results of the low-level LFB analyses must be within +50 of the true value.
            Results of the medium and high-level LFB analyses must be within +30% of the true
            value. If the LFB results do not meet these criteria, then all data for the problem
            analyte(s) must be considered invalid for all samples in the Extraction Batch.

     9.3.4   MS TUNE CHECK - The procedure for conducting the MS Tune Check  is  found in
            Section 10.2.1. Acceptance criteria  for the MS Tune Check are summarized in Sec-
            tion 17, Table 1.  The MS Tune Check must be performed each time a major change
            is made to the mass  spectrometer, and prior to establishing and/or re-establishing an
            initial calibration (Sect. 10.2). Daily DFTPP analysis is not required.

     9.3.5   INTERNAL STANDARDS - The analyst  must monitor the peak areas of the ISs in
            all injections of the Analysis Batch.  The IS response (peak area) in any chromato-

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        graphic run must not deviate from the response in the most recent CCC by more than
        +30%, and must not deviate by more than +50% from the average area measured
        during initial analyte calibration. If an IS area in a chromatographic run does not
        meet these criteria, examine the areas of the ISs in the CCCs, and take corrective
        action such as recalibration, verifying the integrity of the IS solution, and servicing
        the GC/MS system. Reanalyze the extract in a subsequent Analysis Batch.

        9.3.5.1   If the reinjected aliquot produces an acceptable IS response, report results
                from that analysis.

        9.3.5.2  If the reinjected extract fails again, extraction of the sample may need to be
                repeated  provided a sample is available and still within the holding time.
                Otherwise, report results obtained from the reinjected extract, but annotate
                as "suspect/IS recovery."  Alternatively, collect a new sample and
                reanalyze.

9.3.6    SURROGATE RECOVERY - The surrogate analytes are fortified into all field sam-
        ples and QC samples prior to extraction. Calculate the percent recovery (%R) for
        each surrogate using the equation

                                        ( A\
                                 %R=   —  xlOO
                                        UJ

        where A = calculated surrogate concentration for the QC or field sample, and B =
        fortified concentration of the  surrogate analyte.

        9.3.6.1   Surrogate recovery must be in the range of 70 to 130%. When a surrogate
                fails to meet this criterion, evaluate the recovery of the surrogates in the
                CCCs, the integrity  of the CAL solutions, and take corrective action such as
                recalibration and servicing the GC/MS system.  Reanalyze the extract in a
                subsequent Analysis Batch.

        9.3.6.2  If the extract reanalysis meets the surrogate recovery criterion, report only
                data for the reanalyzed extract.

        9.3.6.3   If the extract reanalysis fails the 70 to 130% recovery criterion after
                corrective action, extraction of the sample must be repeated provided a
                sample is available and still within the holding time. If the re-extracted
                sample also fails the recovery criterion, report all data for that sample as
                "suspect/surrogate recovery."  Alternatively, collect a new sample and
                reanalyze.

9.3.7    LABORATORY FORTIFIED SAMPLE MATRIX - A minimum of one LFSM is
        required in each Extraction Batch.  The native concentrations of the analytes in the
        sample matrix must be determined  in a  separate aliquot and the measured value in
        the LFSM corrected for the native concentrations. If a variety of different sample

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        matrices are analyzed regularly, for example drinking water from groundwater and
        surface water sources, performance data must be collected for each source.

        9.3.7. 1   Prepare the LFSM by fortifying a Field Duplicate with appropriate amounts
                of the analyte PDS (Sect. 7.2.3.2) and DACT PDS (Sect, 7.2.3.1).  Select a
                spiking concentration that is greater than or equal to the native background
                concentration, if known. Selecting a duplicate aliquot of a sample that has
                already been analyzed aids in the selection of an appropriate spiking level.
                If this is not possible, use historical data and rotate through low, medium
                and high calibration concentrations when selecting a fortifying
                concentration.

        9.3.7.2   Calculate the  %R using the equation
                                     C

                A = measured concentration in the fortified sample
                B = measured concentration in the unfortified sample
                C = fortification concentration.

        9.3.7.3   Recoveries for samples fortified at concentrations near or at the MRL
                (within a factor of two times the MRL concentration) must be within +50%
                of the true value. Recoveries for samples fortified at all other
                concentrations must be within +30% of the true value.  If the accuracy for
                any analyte falls outside the designated range, and the laboratory
                performance for that analyte is shown to be in control in the CCCs and the
                LFB, the recovery is judged matrix biased. The result for that analyte in the
                unfortified sample is labeled "suspect/matrix."

        NOTE: In order to  obtain meaningful recovery results, correct the measured values
        in the LFSM and LFSMD for the native levels in the unfortified samples, even if the
        native values are less than the MRL. This situation and the LRB are the only
        permitted uses of analyte results below the MRL.

9.3.8    FIELD DUPLICATE OR LABORATORY FORTIFIED SAMPLE MATRIX
        DUPLICATE - A minimum of one FD or LFSMD is required in each Extraction
        Batch. If method analytes are not routinely observed in field samples, analyze an
        LFSMD rather than an FD.

        9.3.8.1   Calculate the relative percent difference (RPD) for duplicate measurements
                (FDi and FD2) using the equation

                                 FD, -FD
                               (FD, +FD2)/2

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            9.3.8.2  RPDs for Field Duplicates must be <30%.  Greater variability may be
                    observed when Field Duplicates have analyte concentrations that are near
                    or at the MRL (within a factor of two times the MRL concentration). At
                    these concentrations, Field Duplicates must have RPDs that are <50%.  If
                    the RPD for any analyte falls outside the designated range, and the
                    laboratory performance for the analyte is shown to be in control in the CCC
                    and the LFB, the precision is judged matrix influenced. The result from the
                    unfortified sample is labeled "suspect/matrix."

            9.3.8.3  If an LFSMD is analyzed instead of an FD, calculate the RPD for duplicate
                    LFSMs (LFSM and LFSMD) using the equation
                          RPD =
                                    LFSM-LFSMDl
                                  (LFSM + LFSMD)/2
xlOO
            9.3.8.4  RPDs for duplicate LFSMs must be <30%. Greater variability may be
                    observed when fortified LFSMs have analyte concentrations that are near or
                    at the MRL (within a factor of two times the MRL concentration).  LFSMs
                    at these concentrations must have RPDs that are <50%. If the RPD for any
                    analyte falls outside the designated range, and the laboratory performance
                    for the analyte is shown to be in control in the CCC and the LFB, the
                    precision is judged matrix influenced.  The result from the unfortified
                    sample is labeled "suspect/matrix."

     9.3.9   QUALITY CONTROL SAMPLE - A QCS is required if an alternate commercial
            source is available for the method analytes. A QCS must be evaluated as part of the
            IDC (Sect. 9.2.5) and each time new stock standard solutions are prepared. If
            standards are prepared infrequently, analyze a QCS at least quarterly. The QCS must
            be fortified near the midpoint of the calibration range and analyzed as a CCC. The
            acceptance criteria for the QCS are the same as for the mid- and high-level CCCs
            (Sect. 10.3.2).  If the accuracy  for any analyte fails the recovery criterion, check the
            standard preparation process, stock standard sources, and the purity of neat materials
            used to prepare the stock standards to locate and correct the problem.

9.4  METHOD MODIFICATION QC REQUIREMENTS - The analyst is permitted to modify
     the GC column,  GC conditions and MS conditions.  The analyst is not permitted to modify
     sample collection and preservation, sample extraction conditions, or QC requirements of the
     method.

     9.4.1   Each time method modifications are made, the analyst must repeat the procedures of
            the IDC (Sect.  9.2) and verify that all QC criteria can be met in ongoing QC samples
            (Sect. 9.3).
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         9.4.2    The analyst is also required to evaluate and document method performance for the
                 proposed method modifications in real matrices that span the range of waters that the
                 laboratory analyzes.  This additional step is required because modifications that per-
                 form acceptably in the IDC, which is conducted in RW, could fail ongoing method
                 QC requirements in real matrices due to common method interferences.  If, for
                 example, the laboratory analyzes finished waters from both surface and groundwater
                 municipalities, this requirement can be accomplished by assessing precision and
                 accuracy (Sects. 9.2.2 and 9.2.3) in a surface water with moderate to high TOC (e.g.,
                 2 mg/L or greater) and a hard groundwater (e.g., hardness greater than 250 mg/L).

         9.4.3    The results of Sections 9.4.1 and  9.4.2 must be appropriately documented by the
                 analyst and must be independently verified by the laboratory's quality assurance
                 officer prior to analyzing field samples.  When implementing method modifications,
                 it is the responsibility of the laboratory to closely review the results of ongoing QC,
                 and in particular, results associated with the CCCs (Sect. 10.3) and the IS area counts
                 (Sect.  9.3.5). If repeated failures are noted, the modification must be abandoned.

10.  CALIBRATION AND STANDARDIZATION

    10.1 Demonstration and documentation of acceptable analyte calibration is required before
         performing the IDC (Sect. 9.2) and prior to analyzing  field samples.  The MS Tune Check
         and the initial GC/MS calibration should be repeated each time a major instrument
         modification or maintenance is performed.

    10.2 GC/MS INITIAL CALIBRATION - An initial calibration requires establishing proper
         GC/MS conditions, confirming the instrument meets the DFTPP tune check criteria, and the
         preparation and analysis of at least five CALs to determine the calibration curve. Calibration
         must be performed using peak areas and the IS technique. Calibration using peak heights and
         external standard calibration are not permitted.

         10.2.1   MS TUNE/MS TUNE CHECK- Calibrate the mass and abundance scales of the MS
                 with calibration compounds and procedures prescribed by the manufacturer with any
                 modifications necessary to meet tuning requirements. Inject five ng or less of
                 DFTPP (Sect. 7.2.5) into the GC/MS system.  Acquire a mass spectrum that includes
                 data for mass/charge ratio (m/z) 45 to 450. Use a single spectrum of the DFTPP
                 peak, an average spectrum of the three highest points of the peak, or an average
                 spectrum across the entire peak to evaluate the performance of the system.
                 Appropriate background subtraction is allowed; however, the background scan(s)
                 must be chosen from the baseline prior to or after elution of the DFTPP peak.  If the
                 DFTPP mass spectrum does not meet all criteria in Table 1, the MS must be retuned
                 and adjusted to meet all DFTPP criteria before proceeding with the initial calibration.

         10.2.2   GC/MS INSTRUMENT CONDITIONS - Operating conditions used during method
                 development are described below. GC/MS operating conditions are summarized in
                 Table  2 (Sect. 17). Conditions different from those described may be used if the
                 method modification QC criteria in Section 9.4 are met.  Alternate conditions include

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       appropriate GC columns, temperature programs, MS conditions, and injection
       techniques and volume, such as cold on-column and direct injection port liners
       and/or large volume injection techniques.  Commercially available GC inlets must be
       used for these alternate injection techniques.

       10.2.2.1 A narrow bore (0.18-mm i.d.) GC column and a relatively fast temperature
                program can be used in order to minimize run time (total run time during
                method development was 12.2 minutes) without jeopardizing resolution.
                This chromatographic technique requires an MS capable of scanning and
                storing sufficient numbers of spectra per second in order to record the
                recommended number of scans per peak (e.g., at least 5, but 10 to 15 scans
                are recommended). Instruments that are not capable of achieving these fast
                scan rates must employ a larger-diameter column (e.g., 0.25-mm i.d.) In
                addition, instruments that are being used with relatively fast temperature
                programs must have GC oven heaters that are sufficiently powerful enough
                to perform fast temperature ramping.

       10.2.2.2 The method analyte, Atrazine-desethyl-desisopropyl, is the fully
                dealkylated form of the parent and as a consequence is very polar. It is
                often referred to as diaminochlorotriazine (DACT) in the literature. Its
                polar nature and its low solubility in the EtOAc solvent and many GC
                stationary phases make this analyte very difficult to chromatograph.
                Several GC columns of varying polarity were investigated, as were various
                injection volumes.  The recommended column is a mid-polarity column
                that has low bleed and performed well for this challenging analyte.  Solvent
                overloading and/or high concentrations of the 13C-labeled DACT surrogate
                can lead to overloading of the column phase. When this happens, the
                DACT peak is broad and asymmetric.

       10.2.2.3 Many of the triazine degradates are polar and subject to breakdown and/or
                adsorption onto active sites in the inlet. A straight, 2-mm quartz liner was
                determined to be optimal for this method.  Even with this liner, a number of
                analytes exhibited a loss in sensitivity at low concentrations.

10.2.3  CALIBRATION STANDARDS - Prepare a set of at least five CAL standards as de-
       scribed in Section 7.2.3.  The lowest concentration of the CALs must be at or below
       the MRL. Additionally, field samples must be quantified using a calibration curve
       that spans the same concentration range used to collect the IDC data (Sect. 9.2), e.g.,
       analysts are not permitted to use a restricted calibration range to meet the IDC
       criteria and then use a larger dynamic range during analysis of field samples.

10.2.4  CALIBRATION - Calibrate the GC/MS system using peak areas and the IS tech-
       nique. Fit the calibration points with either a linear regression or quadratic
       regression (response vs. concentration).  Weighting may be used. Forcing the
       calibration curve through the origin is not recommended.  Suggested quantitation
                                  523-22

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                 ions and retention times for analytes obtained during method development are listed
                 in Table 3.

         10.2.5   CALIBRATION ACCEPTANCE CRITERIA - The calibration is validated by
                 calculating the concentration of the analytes from each of the analyses used to
                 generate the calibration curve using the regression equations. Calibration points that
                 are 
-------
     QC elements are included (Sect. 9). This section describes the procedure used to extract and
     analyze field and QC samples.

11.2  SAMPLE PREPARATION

     11.2.1  Allow field samples to reach room temperature prior to extraction.  Before
            extraction, mark the level of the sample on the outside of the sample bottle for later
            sample volume determination. If using weight to determine volume (Sect. 11.3.2.7),
            weigh the full sample bottle before extraction.

     11.2.2  Add an aliquot of the SUR PDS to each sample to be extracted. For method
            development work, a 10-|iL aliquot of the three-component SUR PDS (Sect. 7.2.2.1)
            and 10 jiL of the atrazine-desethyl-desisopropyl(13C3) SUR stock (Sect. 7.2.2) were
            added to each 250-mL field and QC sample.

     11.2.3  Fortify LFBs, LFSMs, or LFSMDs, with an appropriate volume of analyte PDS
            (Sect. 7.2.3.2) and the atrazine-desethyl-desisopropyl stock standard (Sect. 7.2.3.1).
            Cap and invert each sample several times to mix.

     11.2.4  Proceed with sample extraction using SPE carbon cartridges.

11.3  CARTRIDGE SPE PROCEDURE

     11.3.1  CARTRIDGE CLEANING AND CONDITIONING - Proper cleaning and
            conditioning of the solid-phase sorbent can have a marked effect on method
            precision and accuracy.

            11.3.1.1 Set up extraction columns on the SPE vacuum manifold. Using low
                    vacuum (approximately 1 to 2 inches Hg), rinse each cartridge with two
                    6-mL aliquots of DCM, aspirating completely.

            11.3.1.2 Rinse each cartridge with a 6-mL aliquot of MeOH, being careful not to let
                    the bed to go dry. Follow this with a 6-mL aliquot of RW, again being
                    careful not to let the bed go dry.

     11.3.2  SAMPLE EXTRACTION

            11.3.2.1 Add an additional 4 mL of RW to each cartridge and  attach the sample
                    transfer lines. This additional volume prevents the SPE cartridge bed from
                    going dry while the dead volume in the transfer lines  is being filled.
                    Extract samples at a cartridge flow rate of approximately 10 mL/minute.

            NOTE: Faster flow rates have not been tested and could cause breakthrough.

            11.3.2.2 Dry the cartridges under high vacuum for 10 seconds. Release the vacuum,
                    then add a 0.25-mL aliquot of MeOH to each cartridge. Draw the MeOH to

                                      523-24

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                     waste with brief vacuum, then dry the cartridges under high vacuum for
                     10 minutes.

            11.3.2.3  Elute the analytes from the cartridges into 15-mL conical tubes with 2 mL
                     of EtOAc followed by two, 6-mL aliquots of 9:1 DCM/MeOH. Allow the
                     cartridge beds to briefly soak in solvent before drawing the solvent through
                     the cartridges.

            11.3.2.4  Dry the eluate by passing it through approximately 3 grams of anhydrous
                     Na2SC>4 collecting  it in a 40-mL centrifuge tube.  (A Pasteur pipette may be
                     used for a drying column.) Pre-rinse the Na2SO4with a 1-mL aliquot of 3:1
                     DCM/EtOAc. Rinse the 15-mL conical tube with another 1-mL aliquot of
                     3:1 DCM/EtOAc and pass this through the anhydrous Na2SC>4 drying
                     column, collecting it in the same 40-mL centrifuge tube.  At this point in
                     the procedure, the dried extracts may be stored overnight in the 40-mL
                     tubes at -10 °C, if desired.

            11.3.2.5  Thermostat the 40-mL tubes at approximately 35 °C in a water bath, and
                     blow down the eluates under a stream of nitrogen gas to less than 1 mL (but
                     no less than 1A mL).

            11.3.2.6  Transfer the concentrated eluates to 1-mL volumetric tubes. Rinse the
                     conical tube with a small volume of EtOAc,  and transfer the rinseate to the
                     volumetric. Add IS solution, and adjust to volume. During method
                     development, a 10-[iL aliquot of the IS PDS  (Sect. 7.2.1.2) was added to
                     each extract. Transfer the extracts to autosampler vials and store in a
                     freezer (<-10°C).

            11.3.2.7  SAMPLE VOLUME OR WEIGHT DETERMINATION - Use a graduated
                     cylinder to measure the volume of water required to fill the original sample
                     bottle to the mark made prior to extraction (Sect. 11.2.1). Determine the
                     volume of each sample to the nearest 2 mL for use in the final calculations
                     of analyte concentration (Sect.  12.3).  If using weight to determine volume,
                     reweigh the empty sample bottle. From the weight of the original sample
                     bottle measured in Section 11.2.1, subtract the empty bottle weight.

11.4  ANALYSIS OF SAMPLE EXTRACTS

     11.4.1  Establish GC/MS operating  conditions equivalent to those summarized in Table 2 of
            Section 17. Confirm that compound separation and resolution are similar to those
            summarized in Table 3 and illustrated in Figure 1 (Sect. 17).

     11.4.2 Establish a valid initial calibration following the procedures outlined in Section 10.2
           or confirm that the calibration is still valid by analyzing a CCC as described in
           Section 10.3.
                                       523-25

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         11.4.3  Analyze aliquots of field samples and QC samples. Analyze CCCs at the required
                frequency alternating concentrations as described in Section 10.3.  The GC/MS condi-
                tions used to acquire the initial calibration must be used for all sample analyses. The
                Laboratory Reagent Blank should be the first sample analyzed after the opening CCC.

         NOTE: Each Analysis Batch must begin with the analysis of a CCC at or below the MRL for
         each analyte that the laboratory intends to report. This is true whether or not an initial
         calibration is analyzed. After 24 hours or 20 field samples, the low-level CCC must be
         repeated to begin a new Analysis Batch.  Do not count QC samples (LRBs, LFBs, FDs,
         LFSMs, LFSMDs) when calculating the frequency of CCCs that are required during an
         Analysis Batch.

12.  DATA ANALYSIS AND CALCULATIONS

   12.1   COMPOUND IDENTIFICATION - Establish an appropriate retention time window for
          each analyte to identify them in QC and field sample chromatograms. Base this assignment
          on measurements of actual retention time variation for each compound in standard solutions
          analyzed on the GC/MS over the course of time.  The suggested variation is plus or minus
          three times the standard deviation of the retention time for each compound for a series of
          injections.  The injections from the initial calibration and from the IDC (Sect. 9.2) may be
          used to calculate the retention time window. However, the experience of the analyst should
          weigh heavily on the determination of an appropriate range.

         12.1.1  At the conclusion of data acquisition, use the same software settings established
                during the calibration procedure to identify peaks of interest in the predetermined
                retention time windows.  Initially, identify an analyte by comparison of its retention
                time with that of the corresponding analyte peak in a recent initial calibration
                standard or CCC.

         12.1.2   Some GC/MS programs use a spectrum matching criterion when collecting data in
                full scan mode based on the comparison of field sample spectra (after background
                subtraction if necessary) to reference spectra in the user-created database.  This
                database must be created prior to conducting the IDC from spectra obtained for a
                mid-level to high-level calibration standard and updated as necessary. If available,
                this feature may be utilized as a secondary identification routine; however, the
                primary criterion must be based on the analyte retention time.

    12.2 COMPOUND CONFIRMATION - In general, all ions that are present above 30% relative
         abundance in the mass spectrum of the user-generated database should be present in the mass
         spectrum of the sample component and should agree within an absolute 20% of the relative
         abundance in the reference spectrum. For example, if an ion has a relative abundance of 30%
         in the standard spectrum, its abundance in the sample spectrum should be in the range of 10
         to 50%. Some ions, particularly the molecular ion, are of special importance, and should be
         evaluated even if they are below 30% relative abundance.
                                           523-26

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    NOTE: Compound identification is more challenging when sample components are not resolved
    chromatographically and produce mass spectra containing ions contributed by more than one
    compound. When GC peaks obviously represent more than one sample component (i.e.,
    broadened peak with shoulder(s) or valley between two or more maxima), appropriate analyte
    spectra and background spectra can be selected by examining individual spectra profiles during the
    peak to determine the characteristic ions.  When analytes co-elute (i.e., only one GC peak is
    apparent), the identification criteria may be met but each analyte spectrum will contain extraneous
    ions contributed by the co-eluting compound.

    12.3 COMPOUND QUANTITATION - Calculate analyte concentrations using the multipoint
         calibration established in Section 10.2. Report only those values that fall between the MRL
         and the highest CAL.

         12.3.1   EXCEEDING CALIBRATION RANGE - The 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 using EtOAc with the appropriate
                 amount of IS added to match the original level, and the diluted extract reinjected.
                 Acceptable surrogate performance must be determined from the undiluted sample
                 extract.  Incorporate the dilution factor into final  concentration calculations. The
                 resulting data must be documented as a dilution, and the reported MRLs must reflect
                 the dilution factor.

         12.3.2   Calculations must use all available digits of precision, but final reported concentra-
                 tions should be rounded to an appropriate number of significant figures (one digit of
                 uncertainty);  this is typically two, and not more than three, significant figures.

         12.3.3   Prior to reporting data, the chromatograms must be reviewed for any incorrect peak
                 identifications or poor integrations.

         123.4   Prior to reporting data, the laboratory is responsible for assuring that QC require-
                 ments have been met and that any  appropriate qualifier is assigned.

13.  METHOD PERFORMANCE

    13.1 PRECISION, ACCURACY AND LOWEST CONCENTRATION MINIMUM
         REPORTING LIMITS - Table 4 presents the DL and LCMRL values obtained at EPA.
         LCMRLs were determined and calculated using a procedure described elsewhere.1  Single
         laboratory precision and accuracy data are  presented for three water matrices: RW (Table 5),
         chlorinated surface water (Table 6), and chlorinated groundwater (Table 7). Figure 1
         displays a representative chromatogram fortified at a high concentration in chlorinated
         groundwater.

    13.2 SECOND LABORATORY EVALUATION - The performance of this method was demon-
         strated by  a second laboratory, with results similar  to those reported in Section 17. The
         authors wish to acknowledge  the work of Ms. Diane Gregg and Ms. Meredith Clarage of the
                                           523-27

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     U.S. EPA Region 6 Laboratory, Houston, TX, and Mr. Frederick Feyerherm of Agilent, Inc.,
     for conducting the second laboratory evaluation.

13.3  SAMPLE STORAGE STABILITY STUDIES - Chlorinated surface water samples, fortified
     with method analytes at 5.0 |ig/L, were preserved and stored over a 28-day period as
     specified in Section 8.  The analyte recovery and the precision of three replicate analyses of
     the stored samples, conducted after 0, 7,  14, 21, and 28 days of storage are presented in
     Section 17, Table 8.  These data were used to determine the 28-day holding time.

13.4  EXTRACT STORAGE STABILITY STUDIES - Extract storage stability studies were
     conducted for extracts obtained from a chlorinated surface water fortified at 5.0 |ig/L.  The
     precision and average analyte recovery of triplicate injections conducted after 0, 7, 14, 21,
     and 28 days of storage are presented in Table 9.  These data were used to confirm the 28-day
     holding time.

13.5  PROBLEM COMPOUNDS

     13.5.1  DACT:  Atrazine-desethyl-desisopropyl or DACT is subject to hydrolysis at both
            high (>9) and low (<5) pH but has acceptable stability in neutral aqueous solutions.
            DACT is also degraded in the presence of free available chlorine, but stable in the
            presence of chloramines. Ammonium acetate mitigates both of these modes of loss
            during sample storage, and has sufficient buffer capacity at 20 mM to buffer finished
            groundwaters with relatively high pH that also have high buffer capacity. The low
            solubility of DACT in many common low-polarity GC phases (e.g., DB-1, DB-5)
            leads to phase overloading and distortion of the DACT peak shape at even modest
            column loadings. The recommendations for the Rtx®-50 GC column as well as a
            0.5-|iL injection volume are based on these factors. Even with this mid-polarity
            phase, coupled with a sub-sized injection volume, it is advisable to lower the
            fortification concentration of the 13C3-DACT surrogate to 4 |ig/L in field samples (or
            1 |j,g/mL in the extract) to minimize peak distortion at higher calibration levels.

     13.5.2  Cyanazine, which is the  last eluting analyte, is prone to degradation in the injection
            port.  It also has the highest LCMRL and more variable recovery than any other
            analyte in this method.  For these reasons, a relatively high spike level of 20 |ig/L for
            the surrogate, Cyanazine-t/5; is recommended.

13.6  MATRIX ENHANCED SENSITIVITY - Method analytes may exhibit "matrix-induced
     chromatographic response enhancement."7"11 That is, compounds susceptible to GC inlet
     adsorption or thermal degradation suffer more breakdown when injected in a "cleaner"
     matrix. The injection of a "dirty" sample extract coats surfaces with matrix components and
     "protects" the problem compounds  from decomposition or adsorption.  As a result, a
     relatively greater response is observed for analytes in sample extracts than in calibration
     solutions.  Compounds that exhibit this phenomenon often give analytical results that exceed
     100% recovery in fortified extracts, especially at low concentrations. Analyte areas in CCCs
     may increase after the injection of several "real samples" compared to injections during a
     calibration sequence at the beginning of an Analysis Batch.  If these symptoms are observed,

                                       523-28

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         more frequent recalibration is recommended.  The analyst may also choose to condition the
         injection port after maintenance or prior to starting an Analysis Batch by injecting a few
         aliquots of a field sample extract. Matrix effects are also mitigated by using a quartz inlet
         liner and by minimizing the liner volume as much as is practical (Sect. 6.15.2).

14. POLLUTION PREVENTION

    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, or on-line at http://www.ups.edu/x7432.xml.

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 regula-
         tions, and that laboratories protect the air, water, and land by minimizing and controlling all
         releases from fume hoods and bench operations. In addition, 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, see the
         publications of the American Chemical Society's Laboratory Environment, Health & Safety
         Task Force on the Internet at http://membership.acs.Org/c/ccs/publications.htm. Additional
         waste management information can be found in "Laboratory Waste Minimization and
         Pollution Prevention," Copyright © 1996 Battelle  Seattle Research Center, which can be
         located at http://www.p2pavs.org/ref/01/text/00779/ch05.htm.

16. REFERENCES

    1. Winslow, S. D.; Pepich, B. V.; Martin, J. I; Hallberg, G. R.; Munch D. I; Frebis, C. P.;
       Hedrick, E. J.; Krop, R. A. Statistical Procedures for Determination and Verification of
       Minimum Reporting Levels for Drinking Water Methods. Environ. Sci. Technol. 2006; 40,
       281-288.

    2. Glaser, J.A.; Foerst, D.L.; McKee, G.D.; Quave, S.A.; Budde, W.L. Trace Analyses for Waste-
       waters.  Environ. Sci. Technol. 1981; 15, 1426-1435.

    3. Center for Disease Control, National Institute for Occupational Safety and Health.  Guidelines,
       Recommendations, and Regulations for Handling Antineoplastic Agents.
       http://www.cdc.gov/niosh/topics/antineoplastic/pubs.htmltfb.

    4. Occupational Exposures to Hazardous Chemicals in Laboratories, 29 CFR 1910.1450,
       Occupational Safety and Health Administration (1990).
                                            523-29

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 5.  Safety in Academic Chemistry Laboratories; American Chemical Society Publication,
    Committee on Chemical Safety; 7th Edition, Washington D.C., 2003.

 6.  Panshin, S. Y.; Carter, D. S.; Bayless, E. R. Analysis of Atrazine and Four Degradation
    Products in the Pore Water of the Vadose Zone, Central Indiana. Environ. Sci. Technol. 2000;
    34,2131-2137.

 7.  Erney, D. R.; Gillespie, A. M.; Gilvydis, D. M.; Poole, C. F. Explanation of the Matrix-
    Induced Chromatographic Response Enhancement of Organophosphorous Pesticides During
    Open Tubular Column Gas Chromatography with Splitless or Hot On-Column Injection and
    Flame Photometric Detection. J. Chromatogr. 1993; 638, 57-63.

 8.  Mol, H. G. J.; Althuizen, M.; Janssen, H.; Cramers, C. A.  Brinkman, U.A.Th.  Environmental
    Applications of Large Volume Injection in Capillary GC Using PTV Injectors. J. High Resol.
    Chromatogr. 1996; 19, 69-79.

 9.  Erney, D. R.; Pawlowski, T. M.; Poole, C. F. Matrix Induced Peak Enhancement of Pesticides
    in Gas Chromatography. J. High Resol. Chromatogr. 1997; 20, 375-378.

10.  Hajslova, J.; Holadova, K.; Kocourek, V.; Poustka, J.; Godula, M.; Cuhra, P.; Kempny, M.
    Matrix Induced Effects:  A Critical Point in the Gas Chromatographic Analysis of Pesticide
    Residues. J. Chromatogr. 1998; 800, 283-295.

11.  Wylie, P.; Uchiyama, M. Improved Gas Chromatographic Analysis of Organophosphorous
    Pesticides with Pulsed Splitless Injection. J. AOACInternational. 1996; 79, 2, 571-577.

12.  Determination of Selected Pesticides and Flame Retardants in Drinking Water by Solid Phase
    Extraction and Capillary Column Gas Chromatography /Mass Spectrometry; U.S. EPA
    Method 527, EPA  815-R-05-005.
                                       523-30

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17. TABLES, DIAGRAMS, FLOWCHARTS, AND VALIDATION DATA
TABLE 1.  ION ABUNDANCE CRITERIA FOR BIS(PENTAFLUOROPHENYL)PHENYL
           PHOSPHINE, (DECAFLUOROTRIPHENYL PHOSPHINE, DFTPPf
Mass (m/z)
68
69
70
197
198
199
365
441
442
443
Relative Abundance Criteria
<2%ofWz69
Present
<2%ofWz69
<2% of m/z 198
Present
5-9% of m/z 198
>1% of base peak
<150%ofWz443
Present
15-24% of m/z 442
Purpose of Checkpoint13
Low-mass resolution
Low-mass resolution
Low-mass resolution
Mid-mass resolution
Mid-mass resolution and sensitivity
Mid-mass resolution and isotope ratio
Baseline threshold
High-mass resolution
High-mass resolution and sensitivity
High-mass resolution and isotope ratio
  These ion abundance criteria have been developed specifically for target compound analysis as
  described in this method. Adherence to these criteria may not produce spectra suitable for
  identifying unknowns by searching commercial mass spectral libraries. If the analyst intends to use
  data generated with this method to identify unknowns, adherence to stricter DFTPP criteria as
  published in previous methods12 is suggested.
  All ions are used primarily to check the mass accuracy of the mass spectrometer and data system,
  and this is the most important part of the performance test.  The three resolution checks, which
  include natural abundance isotope ratios, constitute the next most important part of the performance
  test, followed by the correct setting of the baseline threshold, as indicated by the presence of m/z
  365.
                                          523-31

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TABLE 2.  GC/TOF-MS CONDITIONS (LECO Model Pegasus III, GC/TOF-MS)

Chromatographic Conditions

Column:  Restek Rtx-50, 20 meters x 0.18 mm i.d., 0.20-|im film thickness
Injection Liner:  2-mm quartz
Injection Volume: 0.50  |jL; Syringe:  5.0 jiL
Injection Port Temperature: 260 °C
Split Delay:  0.33 minute
Carrier Gas:  He, 1.0 mL/minute, constant flow, initial head-pressure 27 pounds per square inch
Temperature Program: initial oven temperature of 120 °C hold for 2 minutes, then ramp at
  25 °C/minutes to 250 °C and hold for 5 minutes
Total Run Time:  12.2 minutes


TOF-MS Conditions

Scan Delay:  375 seconds
Scan End: 520 seconds
Mass Scan Range: m/z 45 to 260
Mass Defect: 0
Detector Voltage:  1600  volts
Acquisition Rate:  20 spectra/second
Source Temperature:  200 °C
                                          523-32

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TABLE 3. RETENTION TIMES, SUGGESTED QUANTITATION IONS, AND
        RECOMMENDED INTERNAL STANDARD REFERENCES
Analyte
Atrazine-desethyl-desisopropyl-13C3 (SUR)
Atrazine-desethyl-desisopropyl
Terbuthylazine-desethyl
Atrazine-desethyl-Jy (IS#1)
Atrazine-desethyl
Prometon
Atrazine-desisopropyl-^s (SUR)
Atrazine-desisopropyl
Propazine
Atrazine-t/5 (IS#2)
Atrazine
Terbuthylazine
Simazine-t/io (SUR)
Simazine
Prometryn
Ametryn
Simetryn
Cyanazine-t/s (SUR)
Cyanazine
Peak#
Fig. 1
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
Ret. Time
(minute)
6.44
6.44
6.61
6.62
6.65
6.69
6.71
6.73
6.76
6.84
6.86
6.88
6.90
6.95
7.53
7.64
7.74
8.46
8.49
Quan. Ion
(m/z)
148
145
186
176
172
210
178
173
214
205
200
214
193
201
241
227
213
230
225

1
1
1
(IS#1)
1
2
1
1
2
(IS#2)
2
2
2
2
2
2
2
2
2
                                 523-33

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TABLE 4.  LOWEST CONCENTRATION MINIMUM REPORTING LEVELS (LCMRL) AND
         DETECTION LIMITS (DL)
Analyte
Atrazine-desethyl-desisopropyl
Terbuthylazine-desethyl
Atrazine-desethyl
Prometon
Atrazine-desisopropyl
Propazine
Atrazine
Terbuthylazine
Simazine
Prometryn
Ametryn
Simetryn
Cyanazine
LCMRL Concentration Levels
(HS/L)
0.30, 0.40, 0.60, 0.80, 1.00, 1.20, 1.60
0.40, 0.60, 0.80, 1.00, 1.20, 1.60, 2.00
0.30, 0.40, 0.60, 0.80, 1.00, 1.20, 1.60
0.60, 0.80, 1.00, 1.20, 1.60, 2.00, 3.00
0.40, 0.60, 0.80, 1.00, 1.20, 1.60, 2.00
0.40, 0.60, 0.80, 1.00, 1.20, 1.60, 2.00
0.30, 0.40, 0.60, 0.80, 1.00, 1.20, 1.60
0.40, 0.60, 0.80, 1.00, 1.20, 1.60, 2.00
0.40, 0.60, 0.80, 1.00, 1.20, 1.60, 2.00
0.60, 0.80, 1.00, 1.20, 1.60, 2.00, 3.00
0.60, 0.80, 1.00, 1.20, 1.60, 2.00, 3.00
0.60, 0.80, 1.00, 1.20, 1.60, 2.00, 3.00
0.80, 1.00, 1.20, 1.60, 2.00, 3.00, 4.00
LCMRL
(HS/L)
0.48
0.72
0.60
0.84
0.65
0.68
0.40
0.45
0.60
0.85
0.83
0.78
2.1
DL
(HS/L)
0.13
0.13
0.10
0.54
0.19
0.16
0.12
0.14
0.22
0.24
0.45
0.12
0.69
                                 523-34

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TABLE 5. PRECISION AND ACCURACY OF METHOD ANALYTES FORTIFIED AT
         3.0 jig/L AND 20 ug/L IN REAGENT WATER (N=7 SAMPLES)
Analyte
Atrazine-desethyl-desisopropyl
Terbuthylazine-desethyl
Atrazine-desethyl
Prometon
Atrazine-desisopropyl
Propazine
Atrazine
Terbuthylazine
Simazine
Prometryn
Ametryn
Simetryn
Cyanazine
Atrazine-desethyl-desisopropyl-13C3
(SUR) - 4 |ig/L
Atrazine-desisopropyl-t/5 (SUR) -
8|ig/L
Simazine-dio (SUR) - 8 |ig/L
Cyanazine-ds (SUR) - 20 |ig/L
Fortified Cone. = 3.0 ug/L
(n=7)
Mean %
Recovery
103
108
105
88.2
103
97.9
97.0
101
95.1
92.1
92.8
88.7
96.8
98.3
102
96.9
92.5
Relative
Standard
Deviation
6.0
5.0
2.7
9.0
3.5
4.7
3.8
5.0
5.5
4.9
4.6
8.5
10
5.5
3.8
3.8
2.8
Fortified Cone. = 20 ug/L
(n=7)
Mean %
Recovery
90.2
89.6
93.9
84.4
93.1
94.3
96.0
96.4
104
89.4
89.6
95.3
94.9
103
100
88.3
81.9
Relative
Standard
Deviation
2.2
3.8
2.0
2.5
2.2
1.4
1.8
1.4
2.2
1.8
1.1
4.7
2.5
3.1
2.9
3.2
2.4
                                   523-35

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TABLE 6. PRECISION AND ACCURACY OF METHOD ANALYTES FORTIFIED AT
          3.0 jig/L AND 20 ug/L IN A CHLORINATED SURFACE WATER3 (N=7 SAMPLES)
Analyte
Atrazine-desethyl-desisopropyl
Terbuthylazine-desethyl
Atrazine-desethyl
Prometon
Atrazine-desisopropyl
Propazine
Atrazine
Terbuthylazine
Simazine
Prometryn
Ametryn
Simetryn
Cyanazine
Atrazine-desethyl-desisopropyl-
13C3 (SUR) - 4 |ig/L
Atrazine-desisopropyl-Js (SUR)
-8|ig/L
Simazine-t/io (SUR) - 8 |ig/L
Cyanazine-ds (SUR) - 20 |ig/L
Fortified Cone. = 3.0 jig/L
(n=7)
Mean %
Recovery
97.8
97.8
96.1
97.7
102
101
104
97.6
102
97.7
102
95.4
107
100
106
103
107
Relative
Standard
Deviation
1.4
2.5
2.1
5.9
2.0
3.8
4.4
5.6
4.7
5.6
11
4.3
10
2.6
3.6
4.8
3.8
Fortified Cone. = 20 ug/L
(n=7)
Mean %
Recovery
102
98.9
103
97.7
101
98.2
101
100
100
95.5
95.5
98.9
106
107
101
102
97.4
Relative
Standard
Deviation
2.2
6.7
1.4
1.8
1.2
1.1
1.4
1.6
4.3
0.75
1.3
4.1
4.1
4.1
2.0
3.4
4.2
  Surface water physical parameters:
  organic carbon = 4 mg/L.
pH = 7.3; hardness = 140 mg/L; free chlorine =1.7 mg/L; total
                                        523-36

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TABLE 7.  PRECISION AND ACCURACY OF METHOD ANALYTES FORTIFIED AT
            1.2 ug/L AND 20 ug/L IN A CHLORINATED GROUNDWATER" (N=7 SAMPLES)
Analyte
Atrazine-desethyl-desisopropyl
Terbuthylazine-desethyl
Atrazine-desethyl
Prometon
Atrazine-desisopropyl
Propazine
Atrazine
Terbuthylazine
Simazine
Prometryn
Ametryn
Simetryn
Cyanazine
Atrazine-desethyl-desisopropyl-
13C3 (SUR) - 4 |ig/L
Atrazine-desisopropyl-t/5 (SUR)
-8|ig/L
Simazine-dio (SUR) - 8 |ig/L
Cyanazine-ds (SUR) - 20 |ig/L
Fortified Cone. = 1.2 ug/L
(n=7)b
Mean %
Recovery
88.9
100
97.6
97.1
100
93.8
100
91.4
110
90.5
92.9
81.9
102
84.6
92.2
103
108
Relative
Standard
Deviation
10
7.1
2.6
7.0
3.8
4.3
9.5
4.1
4.3
12
7.0
6.2
8.2
9.2
5.2
2.3
5.9
Fortified Cone. = 20 ug/L
(n=7)
Mean %
Recovery
92.6
92.6
93.8
90.3
93.8
98.0
100
98.9
98.8
97.1
94.3
95.0
97.9
101
98.7
91.7
85.8
Relative
Standard
Deviation
3.7
3.8
4.2
3.8
3.9
1.4
1.5
1.7
2.5
2.3
2.6
3.1
5.3
3.5
3.4
2.2
4.2
a  Ground water physical parameters:
b  Atrazine-desethyl-desisopropyl was
hardness = 325 mg/L; free chlorine = 0.5 mg/L.
spiked at 2.0 |ig/L for the low level.
                                          523-37

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TABLE 8. SAMPLE HOLDING TIME DATA FOR METHOD ANALYTES FORTIFIED AT
          5.0 jig/L IN A CHLORINATED SURFACE WATER* b (N=3 SAMPLES)
Analyte

Atrazine-desethyl-
desisopropyl
Terbuthylazine-
desethyl
Atrazine-desethyl
Prometon
Atrazine-desisopropyl
Propazine
Atrazine
Terbuthylazine
Simazine
Prometryn
Ametryn
Simetryn
Cyanazine
DayO
%R
93.2
101
98.0
93.9
98.7
95.7
96.7
97.1
92.1
94.9
89.6
86.8
98.1
%RSD
4.3
4.4
1.4
5.0
0.4
4.1
1.2
1.7
3.2
5.9
4.7
6.3
4.4
Day 7
%R
95.6
100
99.3
96.1
98.5
100
99.0
97.1
102
103
89.2
99.0
109
%RSD
0.26
3.1
3.2
4.9
1.2
4.8
5.4
5.9
2.7
17
9.2
3.9
4.4
Day 14
%R
97.5
98.4
100
88.5
96.3
97.3
93.2
87.8
100
95.1
83.8
101
96.4
%RSD
5.1
6.6
1.6
7.0
1.7
8.5
5.9
5.2
7.6
5.4
11
2.3
0.80
Day 21
%R
95.5
101
98.3
91.7
101
91.7
91.5
93.6
96.4
94.9
93.0
91.6
100
%RSD
9.1
1.5
1.8
13
2.2
1.5
2.2
8.0
2.9
8.2
1.0
2.1
10
Day 28
%R
95.1
104
95.9
94.1
92.4
90.6
91.3
98.0
88.6
94.4
92.0
89.5
99.0
%RSD
5.8
3.5
4.8
2.1
2.5
2.9
4.6
2.1
6.1
5.7
2.9
3.5
8.3
   Surface water physical parameters: pH =
   %R = percent recovery; %RSD = percent
7.4; hardness = 140 mg/L; free chlorine = 2.2 mg/L.
relative standard deviation.
                                         523-38

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TABLE 9. EXTRACT HOLDING TIME DATA FOR METHOD ANALYTES FORTIFIED AT
          5.0 jig/L IN A CHLORINATED SURFACE WATER" b (N=3 SAMPLES)
Analyte

Atrazine-desethyl-
desisopropyl
Terbuthylazine-desethyl
Atrazine-desethyl
Prometon
Atrazine-desisopropyl
Propazine
Atrazine
Terbuthylazine
Simazine
Prometryn
Ametryn
Simetryn
Cyanazine
Atrazine-desethyl-
desisopropyl-13C3 (SUR)
Atrazine-desisopropyl-t/s
(SUR)
Simazine-6?io (SUR)
Cyanazine-^s (SUR)
DayO
%R
93.2
101
98.0
93.9
98.7
95.7
96.7
97.1
92.1
94.9
89.6
86.8
98.1
100
103
95.9
101
%RSD
4.3
4.4
1.4
5.0
0.40
4.1
1.2
1.7
3.2
5.9
4.7
6.3
4.4
4.1
6.2
0.10
3.6
Day 7
%R
91.9
95.9
100
98.5
97.4
102
100
101
100
102
97.7
98.4
97.7
99.3
103
97.4
97.9
%RSD
3.7
2.3
2.0
0.46
1.2
7.5
2.6
2.2
3.4
0.88
4.6
0.29
0.31
7.5
6.4
1.3
1.7
Day 14
%R
97.6
94.0
99.4
93.1
94.8
96.9
97.1
100
97.6
93.1
97.2
97.8
102
104
107
99.4
103
%RSD
2.6
2.1
3.3
4.5
1.4
2.0
1.8
5.7
3.1
9.5
4.8
0.84
8.4
3.2
2.3
2.1
6.2
Day 21
%R
95.5
95.9
96.0
94.3
101
94.5
95.7
95.8
99.2
104
98.6
94.5
108
96.2
103
100
103
%RSD
2.5
6.4
2.3
6.5
3.0
1.7
1.7
11
1.6
5.0
6.2
1.0
8.4
5.2
5.3
0.37
4.0
Day 28
%R
94.8
102
95.3
93.7
94.2
90.3
93.4
98.8
97.0
100
93.9
94.5
102
100
98.8
97.6
104
%RSD
1.9
2.7
1.9
2.5
1.9
4.2
1.6
5.3
3.1
3.4
10.
6.6
7.2
6.5
0.87
1.8
4.0
   Surface water physical parameters: pH = 7.4; hardness = 140 mg/L; free chlorine = 2.2 mg/L.
   %R = percent recovery; %RSD = percent relative standard deviation.
                                         523-39

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TABLE 10.  INITIAL DEMONSTRATION OF CAPABILITY (IDC) QUALITY CONTROL
          REQUIREMENTS
Method
Reference
Section
9.2.1
Section
9.2.2
Section
9.2.3
Section
9.2.4
Section
9.2.5
Requirement
Demonstration of low
system background
Demonstration of
precision
Demonstration of
accuracy
MRL confirmation
Quality Control
Sample (QCS)
Specification and
Frequency
Analyze a Laboratory
Reagent Blank (LRB) prior to
any other IDC steps.
Extract and analyze four to
seven replicate Laboratory
Fortified Blanks (LFBs)
fortified near the midrange
concentration.
Calculate average recovery
for replicates used in Section
9.2.2.
Fortify, extract, and analyze
seven replicate LFBs at the
proposed MRL concentration.
Calculate the mean and the
half range (HR). Confirm
that the upper prediction
interval of results (PIR) and
lower PIR (Sect. 9.2.4.2)
meet the recovery criteria.
Analyze mid-level QCS
sample.

Demonstrate that all method analytes
are  50%
Results must be 70-130% of the true
value.
                                  523-40

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TABLE 11.  ONGOING QUALITY CONTROL REQUIREMENTS
   Method
  Reference
    Requirement
 Specification and Frequency
       Acceptance Criteria
 Section 8.4
Sample holding time
28 days when processed and
stored according to Sections 8.1
and 8.3.
Sample results are valid only if
samples are analyzed within the
sample holding time.	
 Section
 9.3.1
Laboratory Reagent
Blank (LRB)
One with each Extraction Batch
Demonstrate that all method
analytes are below 1/3 the
Minimum Reporting Level (MRL),
and that possible interference from
reagents and glassware do not
prevent identification and
quantitation of method analytes.
 Section
 9.3.3
Laboratory Fortified
Blank (LFB)
One with each Extraction Batch
                                    Rotate the fortified
                                    concentrations between low,
                                    medium, and high levels.
Results of LFB analyses at medium
and high fortifications must be 70-
130% of the true value for each
analyte and surrogate. Results of
the low-level LFB must be 50-
150% of the true value.
 Section
 9.3.5
Internal standard (IS)
Atrazine-c/s and Atrazine-
desethyl-Jy are added to all
standards and sample extracts.
Peak area counts for all ISs in field
and QC sample extracts must be
within ±50% of the average peak
area calculated during the initial
calibration and ±30% from the most
recent continuing calibration check
(CCC).  If ISs do not meet these
criteria, corresponding method
results are invalid.
 Section
 9.3.6
Surrogates
Surrogates are added to all field
and QC samples prior to
extraction.
70-130% recovery
 Section
 9.3.7
Laboratory Fortified
Sample Matrix
(LFSM)
One LFSM per Extraction
Batch. Fortify the LFSM with
method analytes at a
concentration close to but
greater than the native
concentrations (if known).
For LFSMs fortified at
concentrations <2 x MRL, the result
must be within 50-150% of the true
value.  All other LFSMs must be
within 70-130% of the true value.
 Section
 9.3.8
Laboratory Fortified
Sample Matrix
Duplicate (LFSMD)
or
Field Duplicate (FD)
One LFSMD or FD with each
Extraction Batch
For LFSMDs or FDs, RPDs must
be <30% at middle and high levels
of fortification and <50% at
concentrations <2 x the MRL.
 Section
 9.3.9
Quality Control
Sample (QCS)
Analyze mid-level QCS sample
when new stock standard
solutions are prepared and at
least quarterly if stock
standards are prepared less
frequently.	
Results must be 70-130% of the
true value.
                                               523-41

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  Method
 Reference
    Requirement
 Specification and Frequency
       Acceptance Criteria
Section
10.2
Initial calibration
Use the internal standard
calibration technique to
generate a linear or quadratic
calibration curve.  Use at least 5
standard concentrations.
Validate the calibration curve
as described in Section 10.2.5.
When each calibration standard is
calculated as an unknown using the
calibration curve, the lowest level
standard must be within 50-150%
of the true value. All other points
must be within 70-130% of the true
value.
Section
10.3
ccc
Verify initial calibration by
analyzing a low-level CCC at
the beginning of each Analysis
Batch. Subsequent CCCs are
required after every 10 field
samples,  and after the last field
sample in a batch.
The lowest level CCC must be
within 50-150% of the true value.
All other points must be within 70-
130% of the true value.

Results for field samples that are
not bracketed by acceptable CCCs
are invalid.
                                               523-42

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                          12  14
              3,4 5
                                                       15   16    17
                                                                                                 19
             6.66667
                                                                           8
Figure 1. Example chromatogram recorded on the GC/TOF-MS for a chlorinated groundwater fortified with Method 523
         analytes at 5 (J,g/L. Numbered peaks are identified in Table 3.
                                                       523-43

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