EPA815-R-05-009
METHOD 314.1    DETERMINATION OF PERCHLORATE IN DRINKING WATER USING
                 INLINE COLUMN CONCENTRATION/MATRIX ELIMINATION ION
                 CHROMATOGRAPHY WITH SUPPRESSED CONDUCTIVITY
                 DETECTION
                                    Revision 1.0
                                     May 2005
Herbert P. Wagner (Lakeshore Engineering Services, Inc.)
Barry V. Pepich (Shaw Environmental, Inc.)
Chris Pohl, Douglas Later, Robert Joyce, Kannan Srinivasan, Brian DeBorba, Dave Thomas, and
Andy Woodruff (Dionex, Inc., Sunnyvale, CA)
David J. Munch (U.S. EPA, Office of Ground Water and Drinking Water)
                          TECHNICAL SUPPORT CENTER
               OFFICE OF GROUND WATER AND DRINKING WATER
                  U. S. ENVIRONMENTAL PROTECTION AGENCY
                             CINCINNATI, OHIO 45268
                                      314.1-1

-------
                                                                           EPA815-R-05-009
                                      METHOD 314.1

   DETERMINATION OF PERCHLORATE IN DRINKING WATER BY INLINE COLUMN
      CONCENTRATION/MATRIX ELIMINATION ION CHROMATOGRAPHY WITH
                       SUPPRESSED CONDUCTIVITY DETECTION

1.  SCOPE AND APPLICATION

   1.1   This is a sample pre-concentration, matrix elimination ion chromatographic (1C) method using
         suppressed conductivity detection for the determination of perchlorate in raw and finished
         drinking waters. This method requires the use of a confirmation column to validate all
         perchlorate concentrations reported at or above the MRL on the primary column. Precision
         and accuracy data have been generated for perchlorate, with both the primary and
         confirmation columns, in reagent water, finished groundwater, surface water and a Laboratory
         Fortified Synthetic Sample Matrix (LFSSM).  The single laboratory Lowest Concentration
         Minimum Reporting Level (LCMRL) has also been determined in reagent water.l


                                                      Chemical Abstract Services
            Analvte                                   Registry Number (CASRN)
            Perchlorate                                        14797-73-0
   1.2   The Minimum Reporting Level (MRL) is the lowest analyte concentration that meets Data
         Quality Objectives (DQOs) that are developed based on the intended use of this method. The
         single laboratory LCMRL is the lowest true concentration for which the future recovery is
         predicted to fall between 50 and 150 percent recovery with 99% confidence. The single
         laboratory LCRML for perchlorate was 0.140 and 0.130 ug/L for the AS16 and AS20
         columns, respectively.  The procedure used to determine the LCMRL is described elsewhere.l

   1.3   Laboratories using this method will not be required to determine the LCMRL, but will need to
         demonstrate that their laboratory MRL for this method meets the requirements described in
         Section 9.2.4.

   1.4   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 for
         perchlorate is dependent on sample matrix, fortification concentration, and instrument
         performance. Determining the DL for perchlorate in this method is optional (Sect. 9.2.7).
         The reagent water DL for the perchlorate was calculated to be 0.03 ug/L using 7 replicates of
         a 0.10 |ig/L fortification level with the AS 16 columns and 0.03 ug/L for the AS20 columns.
         These values are also provided in.-

   1.5   This method is intended for use by analysts skilled in the operation of 1C instrumentation, and
         the interpretation of the associated data.
                                           314.1-2

-------
                                                                            EPA815-R-05-009
2.  SUMMARY OF METHOD

   2.1   Water samples are collected in the field using a sterile filtration technique. The sample,
         without cleanup, is concentrated onto the concentrator/trap column, which is placed in the
         sample loop position and binds perchlorate more strongly than other matrix anions. The
         sample matrix anions are rinsed from the concentrator column with 1 mL of 10 mM NaOH.
         This weak rinse solution allows the concentrator to retain the perchlorate while eluting the
         majority of the matrix anions, which are directed to waste. The concentrator column is
         switched in-line and the perchlorate is eluted from the concentrator column with a 0.50 mM
         NaOH solution.  Following elution from the concentrator, the perchlorate is refocused onto
         the front of the guard column.  The eluent strength is then increased to 65 mM NaOH which
         elutes the perchlorate from the guard column and onto the analytical column where
         perchlorate is separated from other anions and remaining background interferences. The
         sample loading and matrix elimination steps must use the same eluent flow direction as the
         elution and analytical separation steps. Perchlorate is subsequently detected using suppressed
         conductivity and is quantified using an external standard technique. Confirmation of any
         perchlorate concentration reported at or above the MRL on the primary column is
         accomplished with a second analytical column that has a dissimilar separation mechanism.

3.  DEFINITIONS

   3.1    ANALYSIS BATCH - A sequence of field samples, which are analyzed within  a 30-hour
          period and include no more than  20 field samples. An Analysis Batch must also include all
          required QC samples, which do not contribute to the maximum field sample total of 20.  For
          this method, the required QC samples include:

                Laboratory Synthetic Sample Matrix Blank (LSSMB)
                Continuing Calibration Check  (CCC)
                Laboratory Fortified Synthetic Sample Matrix (LFSSM) CCC Standards
                Laboratory Fortified Sample Matrix (LFSM)
                Laboratory Duplicate (LD) or a Laboratory Fortified Sample Matrix Duplicate
                (LFSMD).

   3.2    ANALYTE FORTIFICATION SOLUTIONS (AFS) - The Analyte Fortification Solutions
          are prepared by dilution of the Analyte Secondary Dilution Solutions (SDS) and are used to
          fortify the LFSMs and the LFSMDs with perchlorate.  It is recommended that multiple
          concentrations be prepared so that the fortification levels can be rotated or adjusted to the
          concentration of target analyte in the  native samples.

   3.3    CALIBRATION BLANK (CB) - An aliquot of reagent water or other blank matrix that is
          treated exactly as a CCC.  The CB is  not sterile filtered and is used to determine if the
          method analyte or other interferences are present in the laboratory environment,  the reagents,
          or the apparatus during the IDC calibration.

   3.4    CALIBRATION STANDARD (CAL) - A solution of the target analyte prepared from the
          Perchlorate Primary Dilution Solution or Perchlorate Stock Standard Solution. The CAL
          solutions are used to calibrate the instrument response with respect to analyte concentration.

                                            314.1-3

-------
                                                                       EPA815-R-05-009
3.5     CONTINUING CALIBRATION CHECK STANDARD (CCC) - A calibration check
       standard containing the method analyte which is analyzed periodically throughout an
       Analysis Batch, to verify the accuracy of the existing calibration for that analyte.

3.6     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. 9.2.7), and accurate quantitation is not
       expected at this level.2

3.7     LABORATORY DUPLICATES (LDs) - Two sample aliquots (LDi and LD2), from a single
       field sample bottle, and analyzed separately with identical procedures. Analyses of LDi and
       LD2 indicate precision associated specifically with laboratory procedures by removing
       variation contributed from sample collection, preservation, and storage procedures.

3.8     LABORATORY FORTIFIED BLANK (LFB) - An aliquot of reagent water or other blank
       matrix to which a known quantity of the method analyte is added. The LFB is analyzed
       exactly like a sample, including the preservation procedures in Section 8.1. Its purpose is to
       determine whether the methodology is in control, and whether the laboratory is capable of
       making accurate and precise measurements.

3.9     LABORATORY FORTIFIED SAMPLE MATRIX (LFSM) - An aliquot of a field sample to
       which a known quantity of the method analyte is added. The LFSM is processed and
       analyzed exactly like a field sample, and its purpose is to determine whether the field sample
       matrix contributes bias to the analytical results. The background concentration of the analyte
       in the field sample matrix must be determined in a separate aliquot  and the measured value in
       the LFSM corrected for native concentrations.

3.10   LABORATORY FORTIFIED SAMPLE MATRIX DUPLICATE (LFSMD) - A second
       aliquot of the field sample used to prepare the LFSM, which is fortified and analyzed
       identically to the LFSM.  The LFSMD is used instead of the Laboratory Duplicate to assess
       method precision and accuracy when the occurrence of the target analyte is infrequent.

3.11   LABORATORY FORTIFIED SYNTHETIC SAMPLE MATRIX (LF S SM) - Aliquots of the
       LSSM which are fortified with perchlorate (Sect. 7.2.2). These QC samples are used, during
       an Analysis Batch, to confirm the integrity of the trapping efficiency of the concentrator
       column and that the analyst has adequate resolution between the common anions and
       perchlorate in high ionic matrices. The LFSSM samples are treated like the  CCCs and are
       not sterile filtered.

3.12   LABORATORY REAGENT BLANK (LRB) - An aliquot of reagent water or other blank
       matrix that is treated exactly as a sample including exposure to all filtration equipment,
       storage containers and internal standards. The LRB is used to determine if the method
       analyte or other interferences are present in the laboratory environment, the reagents, or the
       apparatus.

3.13   LABORATORY SYNTHETIC SAMPLE MATRIX (LSSM) - An  aliquot of reagent water
       that is fortified with 1000 mg/L of chloride, bicarbonate and sulfate. This solution is
       representative of a drinking water containing 3000 mg/L of common anions.
                                        314.1-4

-------
                                                                      EPA815-R-05-009
3.14  LABORATORY SYNTHETIC SAMPLE MATRIX BLANK (LS 8MB) - An aliquot of the
      LSSM that is processed like a field sample and is used to determine if the method analyte or
      other interferences are present in the LSSMSS solution. It is also used to determine whether
      the methodology is in control in terms of low system background.

      NOTE: The LSSMB is processed through all sample collection steps outlined in Section
      8.1. The LSSMB must be sterile filtered.

3.15  LABORATORY SYNTHETIC SAMPLE MATRIX FORTIFICATION SOLUTION
      (LSSMFS) - A dilution of the LSSMSS is prepared to facilitate the addition of sodium to all
      field samples in an accurate manner without necessitating volume correction (Sect. 7.2.3).

3.16  LABORATORY SYNTHETIC SAMPLE MATRIX STOCK SOLUTION (LSSMSS) - The
      LSSMSS contains the common anions chloride, sulfate and bicarbonate at 25.0 g/L. This
      solution is used in the preparation of all CAL and QC samples (Sect. 7.2.2).

3.17  LOWEST CONCENTRATION MINIMUM REPORTING LEVEL (LCMRL) - The single-
      laboratory LCMRL is the lowest true concentration for which the future recovery is predicted
      to fall between 50 and 150 percent recovery with 99% confidence. l

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

3.19  MINIMUM REPORTING LEVEL (MRL) - The minimum concentration that can be
      reported by a laboratory as a quantified value for the target analyte in a sample following
      analysis.  This defined concentration must meet the criteria defined in Section 9.2 and must
      be no lower than the concentration of the lowest calibration standard for the target analyte.

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

3.21  QUALITY CONTROL SAMPLE (QCS) - A solution containing the method analyte at a
      known concentration that is obtained from a source external to the laboratory and different
      from the source of calibration standards. The QCS is used to verify the calibration
      standards/curve integrity.

3.22  REAGENT WATER (RW) - Purified water which does not contain any measurable quantity
      of the target  analyte or interfering compounds at or above 1/3 the MRL.

3.23  SECONDARY DILUTION STANDARD SOLUTION (SDS) - A solution containing the
      method analyte prepared in the laboratory from the PDS and diluted as needed to prepare
      calibration solutions and other analyte solutions.

3.24  STOCK STANDARD SOLUTION (SSS) - A concentrated solution containing the method
      analyte prepared in the  laboratory using assayed reference materials or purchased from a
      reputable commercial source.
                                       314.1-5

-------
                                                                             EPA815-R-05-009
4.  INTERFERENCES

   4.1   Interferences can be divided into three different categories: (i) direct chromatographic co-
         elution, where an interfering analyte response is observed at very nearly the same retention
         time (RT) as the target analyte; (ii) concentration dependant co-elution, which is observed
         when the response of higher than typical concentrations of the neighboring peak overlaps into
         the retention window of the target analyte; and (iii) ionic character displacement, where
         retention times may significantly shift due to the influence of high ionic strength matrices
         (high mineral content or Total Dissolve Solids) overloading the exchange sites on the column
         and significantly shortening the target analyte's retention time.

       4.1.1  A direct chromatographic co-elution may be solved by changing columns, eluent
             strength, modifying the eluent with organic solvents (if compatible with 1C columns),
             changing the detection systems, or selective removal of the interference with
             pretreatment.  Sample dilution will have little  to no effect. The analyst must verify that
             these changes do not induce any negative affects on method performance by repeating
             and passing all the QC criteria as described in Section 9.2.

       4.1.2  Sample dilution may resolve some of the difficulties if the interference is the result of
             either concentration dependant co-elution or ionic character displacement, but it must be
             clarified that sample dilution will alter your MRL by a proportion equivalent to that of
             the dilution. Therefore, careful consideration of DQOs should be given prior to
             performing  such a dilution.

   4.2   Method interferences may be caused by contaminants in solvents, reagents (including reagent
         water), sample bottles and caps, and other sample processing hardware that lead to discrete
         artifacts and /or elevated baselines in the chromatograms.  All items such as these must be
         routinely demonstrated to be free from interferences (less than l/j the perchlorate MRL) under
         the conditions of the analysis by analyzing LRBs and LSSMBs as described in Section 9.2.1.
         Subtracting blank values from  sample results is not permitted.

   4.3   Matrix interferences  may be 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. Water samples high in organic carbon or TDS may have elevated
         baselines or interfering peaks.

   4.4   Equipment used for sample collection and storage has the potential to introduce interferences.
         The potential for interferences from these devices must be investigated during the Initial
         Demonstration of Capability (Sect. 9.2) by preparing and analyzing a LRB and LSSMB. This
         procedure should be repeated each time that a new brand or lot of devices are used to ensure
         that contamination does not hinder analyte identification and quantitation.

   4.5   This method utilizes  a confirmation column that has a separation mechanism that is
         sufficiently different for the primary column  so that perchlorate may be confirmed. The
         suggested primary column, the lonPac AS 16, has a column chemistry that is based on a low
         cross-link vinyl aromatic quaternary monomer.  It was  designed to provide good
         chromatographic performance for polarizable inorganic anions such as perchlorate with
         moderate concentration hydroxide eluents. Although less polarizable than inorganic species
                                             314.1-6

-------
                                                                           EPA815-R-05-009
         such as perchlorate, such aromatic species show enhanced retention due to interaction with the
         pi electrons of the aromatic backbone.  The suggested confirmation column, the lonPac AS20,
         has a column chemistry that is based on a cross-linked quaternary condensation polymer
         completely free of any pi electron containing substituents. As such, it exhibits selectivity for
         polarizable anions which is complementary to the AS 16, but because of the absence of any pi
         electron character, retention of aromatic anionic species is greatly diminished relative to that
         of the AS 16.

       4.5.1  One component that has been shown by 1C and IC-MS to potentially co-elute with
             perchlorate on the lonPac AS16 column when using EPA Method 314.0 protocols is 4-
             chlorobenzenesulfonic acid (4-C1 BSA).3  As shown in Figure 1, with EPA Method
             314.1 protocols, there is some resolution of the two components on the AS16 column and
             the lonPac AS20 column provides excellent separation of perchlorate and 4-C1 BSA.

5.  SAFETY

   5.1   The toxicity  or carcinogenicity of each reagent used in this method has not been precisely
         defined. Each chemical should 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. A reference
         file of MSDSs should be made available to all personnel involved in the chemical analysis.
         Additional references to laboratory safety are available.4"6

6.  EQUIPMENT AND SUPPLIES (References to specific brands or catalog numbers are included
         for illustration only and do not imply endorsement of the product.)

   6.1   NON-STERILE SAMPLE CONTAINERS - 125-mL brown Nalgene bottles (Fisher Cat. No.
         03-313-3C or equivalent).

   6.2   STERILE SAMPLE CONTAINERS - 125-mL sterile high-density polyethylene (HOPE)
         bottles (I-Chem 125-mL sterile HOPE bottle, Fisher Cat. No. N411-0125 or equivalent).

   6.3   SAMPLE FILTERS - Sterile sample filters (Corning 26-mm surfactant free cellulose acetate
         0.2-um filter, Fisher Cat. No. 09-754-13 or equivalent).  If alternate filters are used they
         should be  certified as having passed a bacterial challenge test. 7 In addition, if alternate filters
         or different lots of the recommended filters are used, they must be tested using a LSSMB and
         a LFSSM  fortified at the MRL as outlined in Section 9.2 to insure that they do not introduce
         interferences or retain perchlorate.

   6.4   SYRINGES  - 20-mL sterile, disposable syringes (Henke Sass Wolf 20 mL Luer lock, Fisher
         Cat. No. 14-817-33  or equivalent).

   6.5   VOLUMETRIC FLASKS - Class A, suggested sizes include 10, 50, 100, 250, 500 and 1000
         mL for preparation of standards and eluents.

   6.6   GRADUATED CYLINDERS - Suggested sizes  include 25 and 1000 mL.

   6.7   AUTO PIPETTES - Capable of delivering variable volumes from 1.0 uL to 2500 uL.
                                           314.1-7

-------
                                                                       EPA815-R-05-009
6.8   ANALYTICAL BALANCE - Capable of weighing to the nearest 0.0001 g.

6.9   ION CHROMATOGRAPHY SYSTEM WITH SUPPRESSED CONDUCTIVITY
      DETECTION (1C) - A Dionex model DX500 1C was used to collect the data presented in this
      method.  Alternative 1C systems can be used provided all the QC criteria listed in Section 9
      are met.  The 1C system must have a thermostatically controlled column heater and be capable
      of operating above room temperature (35 °C) and include an ion chromatographic pump and
      all required accessories including, analytical, concentrator and guard columns,
      chromatography module, eluent generator, compressed gasses, autosampler,  suppressor,
      conductivity detector, and a computer-based data acquisition and control system.
      Additionally, the system must be capable of performing inline sample pre-concentration and
      matrix elimination steps.

   6.9.1   CONCENTRATOR COLUMN - 1C column, 4.0 x 35-mm  (Dionex Cryptand Cl or
          equivalent). Any concentrator column that provides effective retention/trapping and
          eventual release of perchlorate while providing the resolution, peak shape, capacity,
          accuracy, and precision (Sect. 9.2) may be used. However, prior to use, the capacity of
          the concentrator column must be evaluated as per Section 11.4.

   6.9.2   PRIMARY GUARD COLUMN - 1C column, 2.0 x 50-mm (Dionex IonPac®AG16 or
          equivalent). Any column that provides adequate resolution, peak shape, capacity,
          accuracy, and precision (Sect. 9.2) may be used.

   6.9.3   CONFIRMATION GUARD COLUMN - 1C column, 2.0 x 50-mm (Dionex
          IonPac®AG20 or equivalent). Any column that provides adequate resolution, peak shape,
          capacity, accuracy, and precision (Sect. 9.2) may be used. The separation mechanism for
          the confirmation guard column must differ from the primary column.

   6.9.4   PRIMARY ANALYTICAL COLUMN - 1C column, 2.0 x 250-mm (Dionex
          IonPac®AS16 or equivalent). Any column that provides adequate resolution, peak shape,
          capacity, accuracy, and precision (Sect. 9.2) may be used.

   6.9.5   CONFIRMATION ANALYTICAL COLUMN - 1C column, 2.0 x 250-mm (Dionex
          IonPac®AS20 or equivalent). Any column that provides adequate resolution, peak shape,
          capacity, accuracy, and precision (Sect. 9.2) may be used. The separation mechanism for
          the confirmation analytical column must differ from the primary column.

   6.9.6   AUTOSAMPLER - A Dionex AS40 autosampler (or equivalent) is required to perform
          the sample pre-concentration/matrix elimination  steps. The method program must
          include a timing sequence to allow the autosampler to load two sample vials before the
          concentrator column is switched in-line to separate and detect perchlorate.  The first
          sample vial contains the sample (2.0 mL) and the second vial contains the rinse solution
          (1.0 mL of 10 mM NaOH), with the filter  cap raised to signify a rinse vial. The method
          programs for the AS16 and AS20 columns are listed in Table 1 A.  The method timing
          sequence for the methods is listed in Table IB.

   6.9.7   ELUENT GENERATOR - An eluent generator (Dionex EG50 or equivalent) with a
          sodium cartridge (EluGen ® PN 058908 or equivalent) is used to prepare the sodium
                                        314.1-8

-------
                                                                            EPA815-R-05-009
             hydroxide eluent for this method. An equivalent eluent generator may be used and/or
             manually prepared eluents may also be used provided that adequate resolution, peak
             shape, capacity, accuracy, and precision (Sect. 9.2) are obtained.  Care must be exercised
             with manually prepared sodium hydroxide eluents to prevent formation of carbonate in
             the eluent from exposure to the atmosphere, which can dramatically alter the
             chromatography.

             NOTE: The Cryptand concentrator columns use manipulation of column capacity as part
             of the mechanism for separation.  The counter ion in the eluent has a strong influence on
             both the concentrator column capacity and the capacity modification kinetics. In this
             work, sodium is used to establish the optimal capacity of the Cryptand concentrator
             column and consequently a sodium cartridge (Dionex PN 058908 or equivalent) rather
             than a potassium cartridge MUST be used with the EG50.

       6.9.8  ANION SUPPRESSOR DEVICE - The data presented in this method were generated
             using a Dionex Ultra II Anion Self-Regenerating  Suppressor (2-mm ASRS, PN 061562)
             for electrolytic suppression of the eluent.  Equivalent suppressor devices may be utilized
             providing a comparable conductivity MRL and DL are achieved and adequate baseline
             stability is attained as measured by a baseline noise of no more than 5 nS per minute over
             the background conductivity.

             NOTE: The conductivity suppressor was set to perform electrolytic suppression at a
             current setting of 100 mA using the external water mode.  Since unacceptable baseline
             stability was observed on the conductivity detector using the Ultra II ASRS in recycle
             mode, the external water mode must be used

       6.9.9  CONDUCTIVITY DETECTOR - Conductivity cell (Dionex CD20 or equivalent)
             capable of providing data as required in Section 9.2.

       6.9.10 CHROMATOGRAPHY MODULE - A chromatography module (Dionex LC30 or
             equivalent) capable of maintaining the columns, suppressor and conductivity cell at 35 °C
             is required.

       6.9.11 DATA SYSTEM - An interfaced data system such as Dionex, Chromeleon Version 6.0
             (or equivalent) is required to acquire, store, and output conductivity data. The computer
             software should have the capability of processing stored conductivity data by recognizing
             a peak within a given retention time window.  The software must allow integration of the
             peak area of any  specific peak between specified time limits. The software must be able
             to construct a linear regression or quadratic calibration curve, and calculate analyte
             concentrations.

7.   REAGENTS AND STANDARDS

    7.1   REAGENTS - Reagent grade or better chemicals should be used in all tests.  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, provided it is first determined that

                                            314.1-9

-------
                                                                       EPA815-R-05-009
      the reagent is of sufficiently high purity to permit its use without lessening the quality of the
      determination.

   7.1.1   REAGENT WATER (RW) - Purified water which does not contain any measurable
          quantity of the target analyte or interfering compounds at or above 1/3 the perchlorate
          MRL. The purity of the water required for this method cannot be overly emphasized.
          The reagent water used during method development was generated from tap water using
          a Millipore ELIX-3 followed by a Millipore Gradient A10 system. The water should
          contain no particles larger than 0.20 microns.

   7.1.2   ELUENT SOLUTION - Sodium hydroxide eluent concentrations of 0.50, 65 and 100
          mM are automatically prepared using the EG50 eluent generator and/or manually
          prepared (Sect. 6.9.7).

   7.1.3   SODIUM BICARBONATE - (NaHCO3, CASRN 497-19-8) - Fluka Cat. No. 71627 or
          equivalent.

   7.1.4   SODIUM CHLORIDE - (NaCl, CASRN 7647-14-5) - Fisher Cat. No. S-271 or
          equivalent.

   7.1.5   SODIUM SULFATE - (Na2SO4, CASRN 7757-82-6) -Fluka  Cat. No. 71959 or
          equivalent.

7.2   STANDARD SOLUTIONS - When a compound purity is assayed to be 96 percent 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 as an example. Even though stability times for standard  solutions are suggested
      in the following sections, laboratories should use standard QC practices to determine
      when their  standards need to be replaced.

   7.2.1   PERCHLORATE STANDARD SOLUTIONS - Obtain the analyte as a solid standard of
          NaClO4 or as a commercially prepared standard from a reputable standard manufacturer.
          Prepare the Perchlorate Stock and Dilution Solutions as described below.

      7.2.1.1 PERCHLORATE STOCK STANDARD SOLUTION (SSS) (1000 mg/L C1O4") - To
             prepare this solution from a solid NaClO4 standard, weigh out 123.1 mg of NaClO4
             into a 100-mL volumetric flask and dilute to volume with reagent water. When
             stored in opaque, plastic storage bottles, the resulting stock solution may be stable for
             up to one year.

      7.2.1.2 PERCHLORATE PRIMARY DILUTION SOLUTION (PDS) (10.0 mg/L C1O4") -
             Prepare the Perchlorate PDS by adding 1.00 mL of the Perchlorate SSS  to a 100-mL
             volumetric flask and dilute to volume with reagent water. This solution is used to
             prepare the Secondary Dilution Solution, the Perchlorate Fortification Solutions and
             the Calibration Solutions below. When stored in opaque, plastic storage bottles, the
             resulting solution is stable for at least one month.
                                       314.1-10

-------
                                                                 EPA815-R-05-009
7.2.1.3 PERCHLORATE SECONDARY DILUTION SOLUTION (SDS) (1.00 mg/L C1O4')
       - Prepare the 1.00 mg/L Perchlorate SDS by adding 10.0 mL of the Perchlorate PDS
       to a 100-mL volumetric flask and dilute to volume with reagent water.  This solution
       is used to prepare the Perchlorate Fortification Solutions, CAL and CCC Standards
       listed below. When stored in opaque, plastic storage bottles, the resulting solution is
       stable for at least one month.

7.2.1.4 PERCHLORATE FORTIFICATION SOLUTIONS (PFS) (50, 200 and 500 |ig/L) -
       The Perchlorate Fortification Solutions are prepared by dilution of the Perchlorate
       SDS and are used to fortify the Laboratory Fortified Blank (LFB), the Laboratory
       Fortified Synthetic Sample Matrix (LFSSM), the Laboratory Fortified Sample Matrix
       (LFSM) and the Laboratory Fortified Sample Matrix Duplicate (LFSMD) with
       perchlorate. It is recommended that multiple concentrations be prepared so that the
       fortification levels can be rotated or adjusted to the concentration of target analyte in
       the native samples. When stored in opaque, plastic storage bottles, the resulting
       solutions are stable for at least one month. A 20-uL aliquot of each PFS added to a
       2.0-mL sample volume yield a perchlorate concentration of 0.50, 2.0 and 5.0 ug/L,
       respectively.

.2   LABORATORY SYNTHETIC SAMPLE MATRIX STOCK SOLUTION (LSSMSS) -
    Prepare a LSSMSS that contains the common anions chloride, sulfate and bicarbonate at
    25.0 g/L as follows. This solution is used in the preparation of all QC samples. A
    dilution of the LSSMSS is used to fortify all samples (Sect. 7.2.3).

7.2.2.1 Weigh out 3.44 g of NaHCO3, 3.72 g of Na2SO4, and 4.00 g of NaCl (Fluka 1627,
       Fluka 71959, Fisher S-271, respectively or equivalent).  Quantitatively transfer these
       to a 100-mL volumetric flask and dilute to volume using reagent water. This solution
       is used to add  100 mg/L of the LSSM to all blanks, CALs and CCCs and all field
       samples.

       NOTE: EPA Method 314.0 incorporated a synthetic sample matrix containing 1000
       mg/L of chloride, carbonate and sulfate that yielded a pH of approximately 10.
       Method 314.1 uses bicarbonate which yields a pH of approximately 8.6, which more
       closely resembles a finished drinking water. It should be noted that pH 10 carbonate
       is a stronger eluent that could cause break-through of perchlorate on the Cryptand
       concentrator column under conditions listed for this method and should therefore not
       be used to prepare this solution.

.3   LABORATORY SYNTHETIC SAMPLE MATRIX FORTIFICATION  SOLUTION
    (LSSMFS) - As noted in Sect. 11.4, the capacity of the Cryptand concentrator column is
    set with sodium. A dilution of the LSSM  Stock Solution is prepared to facilitate the
    addition of sodium to all field samples in an accurate manner yet without necessitating
    volume correction. Prepare an LSSMFS that contains the common anions chloride,
    sulfate and bicarbonate at 12.5 g/L as follows.

       Add  50.0 mL of the LSSMSS to a  100-mL volumetric flask and dilute to volume
       using reagent water.  The LSSMFS solution is used to add 100 mg/L of the common
       anions to all field samples (17 |jL/2.0 mL of field sample).
                                 314.1-11

-------
                                                                      EPA815-R-05-009

   7.2.4   CALIBRATION BLANK (CB) - Prepare a CB that contains 100 mg/L of the common
          anions to ensure effective trapping of the perchlorate by adding 400 |jL of the LSSMSS
          to 100 mL of RW as indicated in the Table below.  The CB is used only during the
          initial calibration to ensure that no perchlorate or interferences are present in the
          CAL standards (containing 100 [tgfL of the common anions) prior to calibration.
          The CB is not sterile filtered prior to analysis.

   7.2.5   LABORATORY FORTIFIED BLANK (LFB) - Prepare an LFB that contains 100 mg/L
          of the common anions  by adding 400 [iL of the LSSMSS to 100 mL of RW and
          fortifying the LFB with the appropriate volume of perchl orate PDS  or SDS as indicated
          in the Table below. The LFB must be sterile filtered prior to analysis.

   7.2.6   LABORATORY SYNTHETIC SAMPLE MATRIX BLANK (LSSMB) - Prepare the
          LSSMB by adding 4000 [iL of LSSMSS to 100 mL of RW as indicated in the Table
          below.  The LSSMB must be sterile filtered prior to analysis.

7.3    CALIBRATION STANDARDS (CAL) - Prepare a calibration curve from dilutions of the
      Perchlorate PDS, the Perchlorate SDS, and the LSSMSS using a minimum of five
      Calibration Standards, which span the concentration range of interest. The lowest CAL
      standard must be at or below the MRL. An example of the dilutions used to prepare the CAL
      standards used to collect the data in Section 17, are shown in the Table below.

      NOTE: CAL standards are not processed with the sample collection devices or protocols.
      This step must be omitted for the CALs in order to identify any potential losses associated
      with the  sample filtration or collection protocols.

7.4    CONTINUING CALIBRATION CHECK STANDARDS (CCC) - Prepare the CCC
      standards from dilutions of the Perchlorate PDS, the Perchlorate SDS, and the LSSMSS. An
      example of the dilutions used to prepare the CCCs that were used to collect the data in Section
      17 are shown in the Table below.

      NOTE: CCC standards are not processed with the sample collection devices or protocols.
      This step must be omitted for the CCCs in order to identify any potential losses associated
      with the  sample filtration or collection protocols.

7.5    LABORATORY FORTIFIED SYNTHETIC SAMPLE MATRIX CCC STANDARDS -- In
      order to continually monitor the integrity of the trapping efficiency of the concentrator
      column throughout an Analysis Batch, the CCCs are also prepared in a 1000 mg/L common
      anion synthetic matrix. These  solutions are termed Laboratory Fortified Synthetic Sample
      Matrix (LFSSM) CCCs and are analyzed following the normal CCCs during the Analysis
      Batch. An example of the dilutions used to prepare the LFSSM CCCs that were used to
      collect the data in Section 17, are shown in the Table below. LFSSM CCCs are processed
      through all sample collection devices and protocols.
                                       314.1-12

-------
                                                                            EPA815-R-05-009
PREPARATION OF CAL, CCC AND LFSSM CCC STANDARDS
CALand CCC
Levels
CB
LSSMB
CAL 1
CAL 2
CAL 3
CAL 4
CAL 5
CAL 6
Low-CCC
Mid-CCC
High-CCC
Low-LFSSM CCC
Mid-LFSSM CCC
High-LFSSM CCC
Vol. of
C1O4
PDS
(uL)





30
50
100

50
100

50
100
Vol. of
C1O4
SDS
(uL)


30
50
100



50


50


Vol. of
LSSMSS
(uL)
400
4000
400
400
400
400
400
400
400
400
400
4000
4000
4000
Final
Vol. of
Std.
(mL)
100
100
100
100
100
100
100
100
100
100
100
100
100
100
Final
Cone, of
Common
Anions
(mg/L)
100
1000
100
100
100
100
100
100
100
100
100
1000
1000
1000
Final
Cone, of
C1O4
(Hg/L)
0.0
0.0
0.30
0.50
1.00
3.00
5.00
10.0
0.50
5.0
10
0.50
5.0
10
8.  SAMPLE COLLECTION, PRESERVATION, AND STORAGE

   8.1   SAMPLE COLLECTION

       8.1.1  Grab samples must be collected in accordance with conventional sampling practices.8

       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 approximately 3 to 5 minutes).  Collect a
             representative sample from the flowing system using a beaker of appropriate size. Use
             this bulk sample to generate individual samples as needed. A volume of at least 20-mL is
             required for each individual sample.

       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. A
             volume of at least 20-mL of filtered sample is required for each individual sample.

       8.1.4  Once representative samples are obtained (at the time of collection), they must be sterile
             filtered (Sect. 8.1.4.1) to remove any native microorganisms.  Perchlorate is known to be
             susceptible to microbiological degradation by anaerobic bacteria.9  Samples are sterile
             filtered to remove microbes and stored with headspace to reduce the potential for
             degradation by any remaining anaerobic organisms.

          8.1.4.1 Remove a sterile syringe (Sect. 6.4) from its package and draw up approximately 25
                mL of the bulk sample (fill the syringe). Remove a sterile syringe filter (Sect 6.3)
                from its package without touching the exit Luer connection.  Connect the filter to the
                syringe making sure that no water from the syringe drops on the exterior of the filter.
                                            314.1-13

-------
                                                                           EPA815-R-05-009
                Depress the syringe plunger gently and discard the first 3-5 mL.  Open a sterile
                sample container (Sect. 6.2) without touching the interior.  Using gentle pressure,
                pass the sample through the filter into the sample container. During this process do
                not let the syringe or filter make contact with the sample container. Following
                filtration, seal the sample container tightly, label and prepare the container for
                shipment. Syringes and filters are single use items and must be discarded after each
                sample.

   8.2   SAMPLE SHIPMENT AND STORAGE - Field samples must be chilled during shipment and
         must not exceed 10 °C during the first 48 hours after collection. Field samples should be
         confirmed to be at or below 10 °C when they are received at the laboratory.  Field samples
         stored in the lab must be held at or below 6 °C until analysis, but should not be frozen.

   8.3   SAMPLE HOLDING TIMES - Field samples that are collected and stored as described in
         Sections 8.1 and 8.2 may be held for 28 days.

9.  QUALITY CONTROL

   9.1.   Quality Control requirements include the Initial Demonstration of Capability (IDC) and
         ongoing QC requirements that must be met when preparing and analyzing field samples. This
         section describes each QC parameter, their required frequency, and the performance criteria
         that must be met in order to meet EPA data quality objectives. The QC criteria discussed in
         the following sections are summarized in Section 17, Tables 5 and 6. These QC requirements
         are considered the minimum acceptable QC criteria. Laboratories are encouraged to institute
         additional QC practices to meet their specific needs.

      9.1.1  METHOD MODIFICATIONS - The analyst is permitted to modify the 1C system,
             columns and separation conditions (Sect. 6.9). However, each time such method
             modifications are made, the analyst must repeat the procedures of the IDC (Sect. 9.2). In
             addition, if an alternate concentrator column is used, the procedure outlined in Section
             11.4 MUST be completed before the IDC is initiated.

   9.2   INITIAL DEMONSTRATION  OF CAPABILITY (IDC) - The IDC must be successfully
         performed prior to analyzing any field samples. Prior to  conducting the IDC, the analyst must
         first generate an acceptable Initial Calibration following the procedure outlined in Section
         10.2. Requirements for the IDC are described in the following sections and are summarized
         in Table 5.

      9.2.1  DEMONSTRATION OF LOW SYSTEM BACKGROUND - Analyze a Laboratory
             Synthetic Sample Matrix Blank (LSSMB) processed through all sample collection steps
             outlined in Section 8.1. The LSSMB must be sterile filtered. Confirm  that the LSSMB
             is reasonably free of contamination and that the criteria in Section 9.3.1 and 9.3.2 are
             met.

             NOTE: It is Good Laboratory Practice to include a blank in the calibration of any
             instrument. As well, the method should be checked for carry-over by analyzing a LSSMB
             blank immediately following the highest CAL standard.  If this LSSMB  sample does not

                                           314.1-14

-------
                                                                     EPA815-R-05-009
       meet the criteria outlined in Section 9.3.1 then carry-over is present and should be
       identified and eliminated.
9.2.2   DEMONSTRATION OF PRECISION - Prepare and analyze 7 replicate LFBs and
       LFSSMs fortified near the midrange of the initial calibration curve. All samples must be
       fortified and processed using the sample collection protocols described in Section 8.1.
       The percent relative standard deviation (%RSD) of the results of the replicate analyses
       must be < 20 percent.

           n. _, nT^   Standard Deviation of Measured Concentrations  ,
           % RSD =	x 100
                                Average Concentration

9.2.3   DEMONSTRATION OF ACCURACY - Using the same set of replicate data generated
       for Section 9.2.2, calculate average recovery. The average recovery of the replicate
       values must be within ± 25 percent of the true value.
                n. _.          Average Measured Concentration  ,nn
                % 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. Establish an initial
       calibration following the procedure outlined in Section 10.2.  The lowest calibration
       standard 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 or validate  the MRL
       following the procedure outlined below.

   9.2.4.1 Fortify and analyze seven replicate Laboratory Fortified Blanks at the proposed MRL
          concentration.  All samples must be fortified and processed using the sample
          collection protocols 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 below.

                                      HRPIR = 3.963S

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


   9.2.4.2 Confirm that the upper and lower limits for the Prediction Interval of Result (PIR =
          Mean j_ HRpiR) meet the upper and lower recovery limits as shown below.

             The Upper PIR Limit must be < 150 percent recovery.

                                           HRP1R    ,
                               FortifiedConcentration

             The Lower PIR Limit must be > 50 percent recovery.
                                     314.1-15

-------
                                                                         EPA815-R-05-009

                                      Mean - HRPIK
                                                PIR     xlOO>50%
                                  FortifiedConcentration

       9.2.4.3 The MRL is validated if both the Upper and Lower PIR Limits meet the criteria
             described above (Sect. 9.2.4.2).  If these criteria are not met, the MRL has been set
             too low and must be determined again at a higher concentration.

   9.2.5  MRL CONFIRMATION IN THE 1000 mg/L LFSSM - Fortify and analyze seven
          replicate LFSSMs fortified at the proposed MRL concentration. All samples must be
          fortified and processed using the sample collection protocols described in Section 8.1.
          Follow the steps outlined in Sections 9.2.4.1 to validate the MRL in the LFSSM. If these
          criteria are not met, the MRL has been set too low and must be determined  again at a
          higher concentration.

   9.2.6  CALIBRATION CONFIRMATION - Analyze a Quality Control Sample as described in
          Section 9.4.1 to confirm the accuracy of the calibration standards/calibration curve.

   9.2.7  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 laboratory to
          determine ifDL determination is required based upon the DQOs.

          Analyses for this procedure should be done over at least 3 days. Prepare at least 7
          replicate fortified LFBs using the sample collection protocols described in Section 8.1.
          Use the solutions described in Section 7.2.1.4 to fortify at a concentration estimated to be
          near the DL. This fortification concentration may be estimated by selecting a
          concentration at 2-5 times the noise level. The DLs in Table 2 were calculated from
          LFBs fortified at 0.10 //g/L. Analyze the seven replicates through all steps of Section 11.

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

          Calculate the DL using the following equation:

                        DL = St(n- 1, 1 - alpha = 0.99)
                 where:
                        t(n-1,1 - alpha = 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.

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

9.3     ONGOING QC REQUIREMENTS - This section  describes the ongoing QC criteria that
       must be followed when processing and analyzing field samples. Table 6 summarizes these
       requirements.

                                        314.1-16

-------
                                                                    EPA815-R-05-009

9.3.1  LABORATORY REAGENT BLANK (LRB) - A LRB (Sect. 3.12) is analyzed during
      the IDC to confirm that potential background contaminants are not interfering with the
      identification or quantitation of perchlorate. If the LRB produces a peak within the
      retention time window of perchl orate that would prevent the determination of perchl orate,
      identify the source of contamination and eliminate the interference before processing
      field samples. Background from the method analyte or other contaminants that interfere
      with the measurement of perchl orate must be below l/j of the MRL.

9.3.2  LABORATORY SYNTHETIC SAMPLE MATRIX BLANK (LSSMB) - A LSSMB
      (Sect. 3.14) is required with each Analysis Batch and is used to confirm that potential
      background contaminants are not in the LFSSM fortification solution and are not
      interfering with the identification or quantitation of perchl orate. If the LSSMB produces
      a peak within the retention time window for perchlorate that would prevent the
      determination of perchlorate, determine the source of contamination and eliminate the
      interference before processing field samples. The LSSMB must contain the LSSM at the
      1000 mg/L concentration and must be sterile filtered. Background contamination must
      be reduced to an acceptable level before proceeding.  Background from the method
      analyte or other contaminants that interfere with the measurement of perchlorate must be
      below l/j of the MRL. Blank contamination may be estimated by extrapolation if the
      concentration is below the lowest calibration standard. This procedure is not allowed for
      field sample results as it may not meet the DQOs. If perchlorate is detected in the
      LSSMB at concentrations equal to  or greater than this level, then all data for perchlorate
      must be considered invalid for all field samples in the Analysis Batch.

9.3.3  CONTINUING CALIBRATION CHECK STANDARDS (CCC) - CCC standards are
      analyzed at the beginning of each Analysis Batch, after every ten field samples, and at the
      end of the Analysis Batch.  See Section 10.3 and Table 6 for concentration requirements
      and acceptance criteria.

9.3.4  LABORATORY FORTIFIED SYNTHETIC SAMPLE MATRIX CCC STANDARDS  -
      CCC standards are prepared in the LFSSM at the same concentration as the CCC
      Standards and analyzed at the same frequency as the CCCs. The LFSSM CCCs are used
      to ensure the integrity of the sample pre-concentration/matrix elimination step and the
      chromatographic separation of perchlorate from other interfering anionic species in very
      high ionic matrices. See Section 10.3 and Table 6 for concentration requirements and
      acceptance criteria.

9.3.5  LABORATORY FORTIFIED BLANK - The LFB is only required during the IDC (Sect.
      9.2) and is not required to be included in the Analysis Batch due to the requirement for a
      LSSMB to be analyzed at the start of each Analysis Batch (Sect. 9.3.2).

9.3.6  LABORATORY FORTIFIED SAMPLE MATRIX (LFSM) - An aliquot of a field
      sample to which a known quantity of the method analyte is added.  The LFSM is
      processed and analyzed exactly like a sample,  and its purpose is to determine whether the
      sample matrix contributes bias to the analytical results.  The background concentration of
      the analyte in the sample matrix must be determined in a separate aliquot and the
      measured value in the LFSM corrected for background concentrations.
                                    314.1-17

-------
                                                                      EPA815-R-05-009

   9.3.6.1 Within each Analysis Batch, a minimum of one field sample is fortified as an LFSM
          for every 20 field samples analyzed.  The LFSM is prepared by spiking a field sample
          with an appropriate amount of the Perchlorate Fortification Solution (Sect. 7.2.1.4).
          The fortification should be delivered in the smallest volume possible to minimize
          dilution of the sample. Select a spiking concentration that is equal to or greater than
          the native concentration, if known. Use historical data and rotate through the
          designated concentrations when selecting a fortifying concentration.

   9.3.6.2 Calculate the  percent recovery (%R) for the analyte using the equation.


                             %R=    "    x 100
            A = measured concentration in the fortified field sample
            B = measured concentration in the unfortified field sample
            C = fortification concentration.

            NOTE: If the fortified concentration is below the native concentration, the fortified
            value is not considered valid. The reported value should be flagged to show that the
            fortification level was lower than native concentration. However, the fortification
            frequency requirement for the method will have been met and the analysis batch
            data considered acceptable.

   9.3.6.3 For field samples fortified at or above their native concentration, recoveries should
          range between 75 and 125 percent, except for low-level fortifications less than or
          equal to the MRL where 50 to 150 percent recoveries are acceptable.  If the accuracy
          of perchlorate falls outside the designated range, and the laboratory performance for
          the analyte is shown to be in control in the CCCs and LFSSM CCCs, the recovery is
          judged to be matrix biased. The result for the analyte in the unfortified field sample
          is labeled suspect/matrix to inform the data user that the results are suspect due to
          matrix effects.

       9.3.6.3.1  Field samples that have an observed positive native perchlorate concentration
                less than the MRL and are fortified at concentrations at or near the MRL
                should be corrected for the native levels in order to obtain meaningful percent
                recovery values.  This is the only permitted use of analyte results below the
                MRL.

9.3.7   LABORATORY DUPLICATE OR LABORATORY FORTIFIED SAMPLE MATRIX
       DUPLICATE (LD or LFSMD) - Within each Analysis Batch, a minimum of one
       Laboratory Duplicate (LD) or Laboratory Fortified Sample Matrix Duplicate (LFSMD)
       must be analyzed. Laboratory Duplicates check the precision associated with laboratory
       procedures.  If target analytes are not routinely observed in field samples, a LFSMD
       should be analyzed rather than a LD.  LFSMDs check the precision associated with
       laboratory procedures.
                                     314.1-18

-------
                                                                       EPA815-R-05-009
       9.3.7.1 Calculate the relative percent difference (RPD) for duplicate measurements (LDi and
             LD2) using the equation.
                                              LD.-LD,
                                                        \   xlOO
                                           (LD1+LD2)/2
       9.3.7.2 RPDs for Laboratory Duplicates should be < 25 percent. Greater variability may be
             observed when Laboratory Duplicates have analyte concentrations that are within a
             factor of 2 of the MRL.  At these concentrations Laboratory Duplicates should have
             RPDs that are < 50 percent. If the RPD of any analyte falls outside the designated
             range, and the laboratory performance for that analyte is shown to be in control in the
             CCCs and LFSSM CCCs, the recovery is judged to be matrix influenced. The result
             for that analyte in the unfortified field sample is labeled suspect/matrix to inform the
             data user that the results are suspect due to matrix effects.

       9.3.7.3 If a LFSMD is analyzed instead of a Laboratory Duplicate, calculate the relative
             percent difference (RPD) for duplicate LFSMs (LFSM and LFSMD) using the
             equation.


                                     LFSM  - LFSMD
                                                          xlOO
                                   (LFSM + LFSMD)/2
       93.7.4 RPDs for duplicate LFSMs should be < 25 percent. Greater variability may be
             observed when LFSMs are fortified at analyte concentrations that are within a factor
             of 2 of the MRL. LFSMs fortified at these concentrations should have RPDs that are
             < 50 percent.  If the RPD of any analyte falls outside the designated range, and the
             laboratory performance for that analyte is shown to be in control in the CCCs and
             LFSSM CCCs, the recovery is judged to be matrix influenced.  The result for that
             analyte in the unfortified field sample is labeled suspect/matrix to inform the data
             user that the results are suspect due to matrix effects.

9.4    QUARTERLY  QC REQUIREMENTS

   9.4.1   QUALITY CONTROL SAMPLES (QCS) - As part of the IDC (Sect. 9.2), each time a
          new Analyte PDS (Sect. 7.2.1.2) is prepared, every time the instrument is calibrated and
          at least quarterly, analyze a QCS sample fortified near the midpoint of the calibration
          range. The  QCS sample should be from a source different than the source of the
          calibration standards.  If a second vendor is not available, then a different lot of the
          standard should be used. The QCS should be prepared and analyzed just like a CCC.
          Acceptance criteria for the QCS is identical to the mid- and high-level CCCs; the
          calculated amount for the analyte must be + 25  percent of the true value.  If measured
          analyte concentrations are not of acceptable accuracy, check the entire analytical
          procedure to locate and correct the problem.
                                        314.1-19

-------
                                                                            EPA815-R-05-009

10.  CALIBRATION AND STANDARDIZATION

    10.1   Demonstration and documentation of acceptable initial calibration for perchlorate is required
          before any field samples are analyzed. If alternative instrumentation and/or concentrator
          columns to those listed in this method are used, the procedure outlined in Section 11.4
          MUST be completed before the calibration can be initiated. If the initial calibration is
          successful, continuing calibration check standards are required at the beginning and end of
          each Analysis Batch, as well as after every tenth field sample.

          NOTE: CAL solutions and CCC standards are not processed with the sample
          collection protocols. This step must be omitted for the CALs and CCCs in order to identify
          any potential losses associated with the sample filtration or collection devices.

    10.2   INITIAL CALIBRATION - Initial calibration is established during the IDC and may be
          reestablished prior to analyzing field samples. However, it is permissible to verify the
          calibration with daily CCCs and LFSSM CCCs. Calibration must be performed using peak
          areas and the external standard technique. Calibration using peak heights is not permitted.

          NOTE: In this method, the CB, LFB,  CAL, QCS and CCC standards are prepared in RW
          fortified with  100 mg/L of the LSSM to ensure optimal trapping of perchl orate. The CB,
          LFB, LRB, CAL, QCS and CCC standards are not sterile filtered. The CB is used only in the
          IDC. On the other hand, the LSSMB represents a  drinking water matrix containing 3000
          mg/L of common anions  and is used in all Analysis Batches and must be sterile filtered.

       10.2.1 INSTRUMENT CONDITIONS - Establish proper operating conditions. Operating
             conditions used during method development are described in Section 17 Table 1A.
             Conditions different from those described may be used if the IDC QC criteria in Section
             9.2 are met.

    10.3   CALIBRATION STANDARDS - Prepare a set of at least  five CAL standards as described
          in Section 7.3. The lowest concentration CAL standard must be at or below the MRL. The
          MRL must be confirmed using the procedure outlined in Section 9.2.4, after establishing the
          initial calibration.

       10.3.1 CALIBRATION - The conductivity detector is calibrated using the external standard
             technique.  Calibration curves may be generated using the 1C data system through the use
             of a first (linear) or second (quadratic) order calibration curves.

       10.3.2 CALIBRATION ACCEPTANCE CRITERIA - The validation of the calibration is
             determined by calculating the concentration of the analyte from each of the analyses used
             to generate the calibration curve.  Each calibration point, except the lowest (< MRL), for
             the analyte should calculate to be  75 to 125 percent of its true value. The lowest point
             should calculate to be 50 to 150 percent of its true value. If these criteria cannot be met,
             the analyst will have difficulty meeting ongoing QC criteria. Corrective action must be
             taken to reanalyze the calibration  standards, restrict the range of calibration, or select an
             alternate method of calibration.

                                            314.1-20

-------
                                                                           EPA815-R-05-009
   10.4   CONTINUING CALIBRATION CHECK (CCC) STANDARDS - The CCCs verify the
          calibration at the beginning and end of each group of analyses, as well as after every 10th
          field sample during analyses. The LRBs, LFBs, LFSSMs, LFSMs, LFSMDs, CCCs and
          LFSSM CCCs are not counted as field samples. The beginning CCCs for each Analysis
          Batch must be at or below the MRL in order to verify instrument sensitivity and the accuracy
          of the calibration curve prior to the analysis of any field samples. Subsequent CCCs should
          alternate between a medium and high concentration.

          NOTE: The analyst may chose to also run a mid-level CCC at the start of an Analysis Batch.

       10.4.1 Inject an aliquot of the CCC standards and analyze with the same conditions used during
             the initial calibration.

       10.4.2 Calculate the concentration of the analyte in the CCC standards. The calculated amount
             for the analyte for medium and high level CCCs must be ± 25 percent of the true value.
             The calculated amount for the lowest CCC level for the analyte must be within ± 50
             percent of the true value.  If these conditions do not exist, then all data for the analyte
             must be considered invalid, and remedial action (Sect. 10.4.4) should be taken which may
             require recalibration.  Any field samples that have been analyzed since the last acceptable
             calibration verification and are still within holding time should be reanalyzed  after
             adequate calibration has been restored.

       10.4.3 LABORATORY FORTIFIED SYNTHETIC SAMPLE MATRIX CCC  STANDARDS -
             As noted in Section 9.3.4, LFSSM CCCs Standards are prepared and analyzed to verify
             the integrity of the concentrator column during the Analysis Batch to ensure that high
             ionic strength drinking water matrices will not exceed the capacity of the concentrator
             column. These QC samples are fortified at the same level and run at the same frequency
             as the CCC Standards and are required to meet the same recovery criteria (Sect. 10.4.2).

       10.4.4 REMEDIAL ACTION - Failure to meet CCC or LFSSM CCC QC performance criteria
             may require remedial action. Maintenance such as confirming the integrity of the
             trapping efficiency of the concentrator column and matrix elimination step or
             regenerating or replacing the 1C columns will require re-calibration (Sect. 10.2).

11.  PROCEDURE

   11.1   Important aspects of this analytical procedure include proper field sample collection and
          storage (Sect. 8.1), ensuring that the instrument is properly calibrated (Sect. 10.2) and that all
          required QC are met (Sect. 9) during the Analysis Batch.  This section describes the
          procedures for field sample preparation and analysis. If alternative instrumentation and/or
          concentrator columns to those listed in this method are used, the procedure outlined in
          Section 11.4 MUST be followed prior to analyzing field samples.

   11.2   SAMPLE PREPARATION

       11.2.1 Collect and store field samples as described in Section 8.1.
                                           314.1-21

-------
                                                                         EPA815-R-05-009
   11.2.2 Process all LSSMBs, LFSMs and LFSMDs using the sample collection protocols is
          Section 8.1.

   11.2.3 Transfer a 2.0-mL aliquot of each field or QC Sample to an autosampler vial.  Add 17 uL
          of the LSSMFS (Sect. 7.2.3) to all field sample autosampler vials.  Place the autosampler
          vial in the appropriate position.

   11.2.4 For each QC standard and field sample to be analyzed, prepare a second autosampler vial
          containing 1.0 mL of the lOmM NaOH rinse solution with the filter cap raised to signify
          a rinse vial. Place the rinse vials in the  autosampler rack, after every QC standard and
          field sample.

11.3    SAMPLE ANALYSIS

   11.3.1 Establish the instrument operating conditions as described in Table 1A of Section 17.
          Confirm that the analyte retention times for the calibration standards are stable.

          NOTE: The ionic strength of the common anion solution used to prepare the LFSSM
          CCCs will cause these solutions to have shorter retention times (see Sect. 11.3.4.1).

   11.3.2 Establish a valid initial calibration following the procedures outlined in Section 10.2 or
          confirm that the calibration is still valid by running a low-level CCC as described in
          Section 10.4.  If establishing an initial calibration for the first time, complete the IDC as
          described in Section 9.2.

   11.3.3 Analyze field and QC samples at their required frequencies using the same conditions
          used to collect the initial calibration. Table 7 shows an acceptable analytical sequence
          that contains all method-required QC samples.

   11.3.4 COMPOUND IDENTIFICATION - Establish an appropriate retention time window for
          perchlorate to identify it in QC and field sample chromatograms.

       11.3.4.1   High ionic strength matrices have the potential to cause an increase in background
                 conductivity and severe tailing as other anions elute from the column and cause
                 the perchlorate retention time to decrease.

                 NOTE:  As a result of the difference in ionic strength of the 100 and 1000 mg/L
                 common anion matrices, the retention time for perchlorate in the 1000 mg/L
                 matrix is approximately 0.2 minutes shorter than in the 100 mg/L matrix (the
                 higher ionic strength matrix may act as a stronger eluent) using the conditions
                 outlined in Table 1 A. Since the ionic strength of drinking water matrices may
                 vary considerably, the RT window for perchlorate must be set wide enough to
                 account for the variability in the ionic strength of the drinking water
                 matrices and yet exclude any potential interfering peaks. A window of
                 approximately 0.4 minutes has been found to be acceptable; however setting the
                 window too wide may require additional analyses on the confirmation column.
                                         314.1-22

-------
                                                                        EPA815-R-05-009
       11.3.4.2  COMPOUND CONFIRMATION - Field samples that have a perchlorate result
                on the primary column at or above the MRL require confirmation with a second
                analytical column that has a dissimilar separation mechanism. EPA Methods
                331.0 and 332.0 can be used for confirmation of perchlorate results obtained using
                EPA Method 314.1.

   11.3.5  EXCEEDING CALIBRATION RANGE - The analyst must not extrapolate above the
          established calibration range.  If an analyte result exceeds the range of the initial
          calibration curve, the field sample may be diluted with reagent water and the diluted field
          sample re-injected (the LSSMFS must be added to the diluted field sample prior to
          analysis). Incorporate the dilution factor into final concentration calculations.  The
          dilution will also affect the perchlorate MRL.

11.4    CONCENTRATOR COLUMN EVALUATION - This method was developed with a
       Dionex Cryptand Cl concentrator column. Alternate columns are allowed, but prior to their
       use, they must be evaluated to optimize sample injection volume, to confirm that the matrix
       elimination step does not remove perchlorate, and to confirm that the perchlorate is
       quantitatively transferred to and refocused on the guard column prior to separation on the
       analytical column.  The entire success of this method is totally dependent upon development
       of column combinations that  accomplish the aforementioned protocols. The procedure is
       challenging and requires very experienced 1C chemists to evaluate alternative concentrator
       column/guard and analytical column combinations using the procedures described below.

   11.4.1  CONCENTRATOR COLUMN CAPACITY DETERMINATION - Any concentrator
          column that provides effective retention/trapping and eventual release of perchlorate
          while providing the resolution, peak shape, capacity, accuracy, and precision (Sect. 9.2)
          may be used. However, prior to use, the capacity of the concentrator column must be
          evaluated. The analyst must demonstrate the ability to load (or  concentrate) at least 2.0
          mL of a 5.0 |J,g/L perchlorate standard in the 1000 mg/L LFSSM (the loading volume
          required to obtain the data presented in this method) without exceeding more than 80% of
          the capacity of the concentrator column.  This requirement ensures that the addition of
          the 1000 mg/L of common anions to the field samples will not exceed the capacity of the
          concentrator column.

          11.4.1.1   Prepare 100-mL of the 5.0 |ig/L LFSSM CCC according to directions in Table
                    1. Load increasing volumes of the LFSSM CCC (1.0, 2.0, 3.0, 4.0 and 5.0-
                   mL, smaller increments may be used if desired) using the procedure outlined
                   for sample preparation and analysis sections (Sect. 11.2, 11.3).  Observe when
                   perchlorate break-through occurs (i.e., no further increase in observed
                   perchlorate peak area or concentration).  Plotting the peak area or
                   concentration versus load volume (as a histogram) will establish the volume at
                   which break-though of the perchlorate becomes evident. At this point, 100%
                   of the capacity of the concentrator column has been exceeded.  Ensure that the
                   load volume to be used does not exceed the 80% restriction. It is
                   recommended that this procedure be reproduced at least twice to confirm the
                   break-through point.
                                        314.1-23

-------
                                                                     EPA815-R-05-009
11.4.2 EVALUATION OF MATRIX ELIMINATION CONDITIONS -  Prior to use of a
      concentrator column other than the one listed in this method, the matrix elimination
      protocols must be evaluated in order to ensure that the perchlorate is retained on the
      concentrator column while the interfering matrix anions are removed (to an acceptable
      level) and sent to waste.

   11.4.2.1   Once the load volume has been established, this can be accomplished by rinsing
             the concentrated perchlorate (on the concentrator column) with different
             concentrations and volumes of rinse solution. Prepare several weak NaOH rinse
             solutions (0.50, 1.0 and 1.5 mM). Prepare several autosampler vials containing
             the optimized volume of the 5.0 |ig/L LFSSM CCC. Prepare several autosampler
             rinse vials containing different volumes of the NaOH rinse solutions (0.50, 1.0
             and 1.5-mL) and analyze using the procedure outlined for sample preparation and
             analysis sections (Sect. 11.2,  11.3).  Choose a concentration and volume that will
             meet the above criteria. The background conductivity must be less than 1.5 |j,S
             when perchlorate elutes in order to obtain data similar to that reported in Tables 2,
             3, and 4.

      11.4.2.1.1  The rinse solution used to collect this data was 1.0 mL of 10 mM NaOH,
                 prepared from 50% NaOH by diluting 0.8 g of 50% NaOH to 1L with RW.  In
                 order to prevent accumulation of carbonate in the rinse solution, the rinse
                 solution is  stored, under helium, in a pressurized vessel fitted with a two-way
                 valve on the out line in order to withdraw the rinse solution as required. This
                 rinse solution is prepared fresh weekly.

11.4.3 EVALUATION OF WASH STEP CONDITIONS - Prior to use of a concentrator
      column other than the one listed in this method, the wash step, which elutes the
      perchlorate off the concentrator column and refocuses it at  the head of the guard column,
      must be evaluated. The wash step ensures quantitative transfer of the concentrated
      perchlorate to the guard column head and minimizes band-broadening by ensuring that
      the perchlorate is efficiently refocused on the guard column before the eluent strength is
      increased to effect separation and detection of the perchlorate on the analytical  column.
      These steps are critical to method performance and were carefully optimized for the
      Cryptand concentrator column during method development.

   11.4.3.1   Once the load volume and rinse solution concentration and volume have been
             established, evaluation of the wash step conditions is accomplished by removing
             the guard and analytical columns from the system and connecting the concentrator
             column directly to the conductivity detector.  Use the EG50 to prepare  the wash
             solutions or use the manually prepared NaOH wash solutions (0.50, 1.0 and 1.5
             mM). Using the optimized load volume and rinse solution determined above, use
             the 5.0 |ig/L LFSSM  CCC and rinse solution and modify the method to allow
             different rinse  times (10, 12 and 15 minutes) and concentrations of wash solution
             to be evaluated using the procedure outlined for sample preparation and analysis
             sections (Sect. 11.2, 11.3). Observe the time at which all the perchlorate has
             eluted from the concentrator column (baseline returned minimum conductance).
             The addition of a couple of minutes will ensure complete removal of perchlorate
             from the concentrator column in all matrices. The 12 minute wash step for this
                                     314.1-24

-------
                                                                            EPA815-R-05-009
                    method provides a non-Gaussian peak that shows when the conductivity baseline
                    has returned to the minimum.

          11.4.3.2   After establishing the optimal load volume, rinse solution concentration and
                    volume and wash time, concentration and volume, analyze a low-level CCC and
                    LFSSM CCC to ensure that the optimal conditions chosen provide acceptable
                    chromatography and peak shape and area for perchlorate.

             11.4.3.2.1 The EG50 was used to prepare the wash solution (0.50 mM NaOH for 12
                       minutes) used to collect the data reported in Tables 2, 3, and 4. If the
                       manually prepared wash solution is used, the same precautions to prevent
                       accumulation of carbonate in the wash solution are required. This wash
                       solution is prepared fresh weekly.

12. DATA ANALYSIS AND CALCULATIONS

   12.1  Identify the analyte present in the field and QC samples as described in Section 11.3.4.

         NOTE: Since the ionic strength of the drinking water matrices can vary dramatically, and
         perchlorate elutes on the trailing edge of the residual anions present in the sample, the
         background conductivity is not the same for all injections. Consequently,  the slope of the
         baseline when perchlorate elutes may vary from sample to sample. As a result, it is quite
         possible that the pre-set, auto- integration parameters may not start and stop peak integration
         the same for every  sample. Therefore, the analyst must thoroughly review all chromatograms
         and some of the chromatograms may require manual integration of the perchlorate peak.

   12.2  Calculate the perchlorate concentrations using the multi-point calibration established in
         Section 10.2.  Quantify only those values that fall between the MRL and the highest
         calibration standard. Field samples with target analyte responses that exceed the highest
         calibration standard require dilution and reanalysis (Sect. 11.3.5).

       12.2.1 As noted in Section 9.3.2, it may be necessary to extrapolate below the MRL to estimate
             contaminants in LRBs and LSSMBs and to correct for native levels of perchlorate below
             the MRL when field samples are fortified at or near the MRL. These are the only
             permitted use of analyte results below the MRL.

   12.3  Calculations must utilize all available digits of precision, but final reported concentrations
         should be rounded to an appropriate number of significant figures (one digit of uncertainty),
         typically two, and not more than three significant figures.

   12.4  Prior to reporting data, the laboratory is responsible for assuring that QC requirements have
         been met or that any appropriate qualifier is documented.

13. METHOD PERFORMANCE

   13.1  PRECISION, ACCURACY AND DETECTION LIMITS - Tables for these data are
         presented in Section 17. Instrumental conditions are presented in Table 1A.  The LCMRL for
         perchlorate with both the AS 16 and AS20 columns is presented in Table 2 and was calculated
                                            314.1-25

-------
                                                                            EPA815-R-05-009
         using a procedure described elsewhere.1 Single laboratory precision and accuracy data are
         presented in Tables 3 and 4.

   13.2  Figure 1 is a representative chromatogram showing the separation of perchlorate from 4-C1
         BSA and Figure 2 shows a chromatogram of a surface and a ground water fortified with 1.0
         Hg/L perchlorate and Figure 3 shows a chromatogram of 3.0 //g/L C1O4" in the 50, 500 and
         lOOOmg/LLSSM.

14. POLLUTION PREVENTION

   14.1  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. up s. edu/community/storeroom/Chemi cal_Wastes/wastearti cl e s. htm.

15. WASTE MANAGEMENT

   15.1  The analytical procedures described in this method generate relatively small amounts of waste
         since only small amounts of reagents are used. The matrices of concern are finished drinking
         water or source water. However, the Agency requires that laboratory waste management
         practices be conducted consistent with all applicable rules and regulations, and that
         laboratories protect the air, water, and land by minimizing and controlling all releases from
         fume hoods and bench operations.  Also, compliance is required with any sewage discharge
         permits and regulations, particularly the hazardous waste identification rules and land disposal
         restrictions. For further information on waste management, consult "The Waste Management
         Manual for Laboratory Personnel" available from the American Chemical Society at the
         address in Section 14.1, or on-line at: http://www.p2pays.org/ref/0l/text/00779/ch 15.htm.

16. REFERENCES

   1.  Revisions to the  Unregulated Contaminant Monitoring Regulation for Public Water Systems,
      Proposed Rule, 2004.

   2.  Glaser, J.A., D.L. Foerst, G.D. McKee, S.A. Quave, and W.L. Budde, "Trace Analyses for
      Wastewaters", Environ. Sci. Technol.. 15 (1981) 1426_1435.

   3.  Personal Communication.

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

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

   6.  "Safety in Academic Chemistry Laboratories," American Chemical  Society Publication,
      Committee on Chemical Safety, 3rd Edition, 1979.
                                           314.1-26

-------
                                                                          EPA815-R-05-009

7.  Blosse, P.T., Boulter, E.M., Sundaram, S., "Diminutive Bacteria Implications for Sterile
   Filtration", Pall Corporation, East Hills, NY.

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

9.  Xu, J., Y. Song, B. Min, L. Steinberg,  and B.E. Logan. 2003. Microbial degradation of
   perchlorate: principles and applications. Environ. Engin. Sci, 20(5): 405-422.
                                         314.1-27

-------
                                                                        EPA815-R-05-009
 17.0 TABLES, DIAGRAMS, FLOWCHARTS, AND VALIDATION DATA

 TABLE LA. INSTRUMENTAL CONDITIONS
Standard Conditions and Equipment for Primary Analyses
Ion Chromatograph:
Pump
Conductivity Suppressor:
Chromatography Module
Detector:

Eluent Generator EG50:
Autosampler:
Columns :
Sample loop:
Load Volume:
Rinse Solution:
Eluent Flow:
Typical System Back-pressure:
Total analysis time:
                            Dionex DX500
                            GP40, 2-mm microbore
                            Dionex 2-mm Ultra II ASRS external water mode, 100 mA
                            Dionex LC30, temperature controlled at 35 °C
                            Dionex CD20 suppressed conductivity detector, background
                            conductivity: 1.0 [j,S
                            0.50, 65 and 100 mMNaOH (see Table IB)
                            Dionex AS40
                            Concentrator column Dionex Cryptand Cl, 4 x 35-mm
                            Guard column Dionex AG16, 2 x 50-mm
                            Analytical column Dionex AS 16, 2 x 250-mm
                            Cryptand Cl concentrator column (b)
                            2.0 mL of sample
                            l.OmLof IQmMNaOH
                            0.25 mL/min
                            2350 psi
                            43 minutes
     (a) Mention of trade names or commercial products does not constitute endorsement or
        recommendation for use.
     (b) See Section 11.4

Standard Conditions and Equipment for Confirmation Analyses ^:
Ion Chromatograph:
Pump
Conductivity Suppressor:
Chromatography Module
Detector:

Eluent Generator EG50:
Autosampler:
Columns :
Sample loop:
Load Volume:
Rinse Solution:
Eluent Flow:
Typical System Back-pressure:
Total analysis time:
                            Dionex DX500
                            GP40, 2-mm microbore
                            Dionex 2-mm Ultra II ASRS external water mode, 100 mA
                            Dionex LC30, temperature controlled at 35 °C
                            Dionex CD20 suppressed conductivity detector, background
                            conductivity: 1.0 [j,S
                            0.50, 65 and 100 mMNaOH (see Table IB)
                            Dionex AS40
                            Concentrator column Dionex Cryptand Cl, 4 x 35-mm
                            Guard column Dionex AG20, 2 x 50-mm
                            Analytical column Dionex AS20, 2 x 250-mm
                            Cryptand Cl concentrator column (b)
                            2.0 mL of sample
                            l.OmLof IQmMNaOH
                            0.25 mL/min
                            2350 psi
                            48 minutes
     (a) Mention of trade names or commercial products does not constitute endorsement or
        recommendation for use.
     (b) See Section 11.4
                                         314.1-28

-------
                                                                      EPA815-R-05-009
  TABLE I.B. TIMING SEQUENCE FOR EPA METHOD 314.1 WITH AS16 and AS20
               COLUMNS
Time
-13.60
-13.599
-5.010
-5.000
0.000


11.999
12.000
24.999
24.500
29.000
30.000
30.000
Eluent
100 mM
100 mM
100 mM
0.50 mM
0.50 mM
0.50 mM
0.50 mM
0.50 mM
65 mM
65 mM
100 mM
100 mM
100 mM
100 mM
Duration
0.01 sec
138 sec




1740 sec







Command
Pump Relay l.Open
Pump Relay 1. Closed
Start change to 0.50
mM
ChangetoO.SOmM
ECD.Autozero
ECD_l.Acqon
Pump Inject Valve. Inj
ectPosition
Start change to 65 mM
Change to 65 mM
Start change to 100
mM
Change to lOOmM
Pump Inject Valve. Lo
adPosition (1740 sec)
BCD l.Acqoff
Wait/end
Function
Initiates the program
Starts the AS40
ChangetoO.SOmM
Establish 0.50 mM in columns
Auto-zero the detector
Start data collection
Switch concentrator column to
inject position, elute perchl orate
off trap & refocus on AG16
Change to 65 mM
Separate & detect perchlorate
Change to lOOmM
Clean columns & establish
capacity of trap column
Switch concentrator column to
load position
Stop data collection
Eluent Generator ready to start
next run
Minor changes for AS20 columns
      Increase the run time to 35 minutes to allow for the fact that perchlorate elutes about 4 minutes
      later on the AS20 column.
                                        314.1-29

-------
                                                                      EPA815-R-05-009
 TABLE 2.  LOWEST CONCENTRATION MRL AND DLs FOR PERCHLORATE
Analytical
Column
AS16
AS20
Analyte
C1O4"
cicv
LCMRLa (ug/L)
0.14
0.13
*DL (ug/L)
0.03b
0.03 b
       aLCMRLs were calculated according to the procedure in reference 1
       *The DL was calculated from data acquired on a single day
       bReplicate fortifications at 0.10 jig/L

TABLE 3. 1C PRECISION AND RECOVERY DATA FOR PERCHLORATE IN VARIOUS
          MATRICES WITH AS16 COLUMNS (n=7)
Matrix
*Reagent Water

Chlorinated Surface Water

Chloraminated Surface Water

Chlorinated Ground Water

***LFSSM

Unfortified
Concentration
(ug/L)
<0.14**
<0.14**
0.63
0.63
<0.14**
<0.14**
<0.14**
<0.14**
<0.14**
<0.14**
Fortified
Concentration
(ug/L)
0.50
5.0
1.0
5.0
1.0
5.0
1.0
5.0
0.50
5.0
Mean %
Recovery
102
90.0
82.6
85.8
83.1
89.3
75.9
92.4
102
80.9
% RSD
2.6
3.2
2.7
2.0
3.6
1.8
5.4
3.3
2.8
1.3
 * Reagent water containing 100 mg/L LSSM.  **The LCMRL = 0.14 ug/L for the AS16 column.
 ***LFSSM Reagent water containing 1000 mg/L LSSM. Described in Section 3.11 and 3.13.

TABLE 4. 1C PRECISION AND RECOVERY DATA FOR PERCHLORATE IN VARIOUS
          MATRICES WITH AS20 COLUMNS (n=7)
Matrix
*Reagent Water

Chloraminated Surface Water

Chlorinated Ground Water

**LFSSM

Unfortified
Concentration
(Ug/L)
<0.13**
<0.13**
<0.13**
<0.13**
0.22
0.22
<0.13**
<0.13**
Fortified
Concentration
(Ug/L)
0.50
5.0
0.50
5.0
0.50
5.0
0.50
5.0
Mean %
Recovery
104
94.2
108
97.8
96.2
98.0
97.4
86.3
% RSD
5.3
1.5
2.2
2.0
9.4
0.70
4.4
1.3
 * Reagent water containing 100 mg/L LSSM.  **The LCMRL = 0.13 ug/L for the AS20 column.
 **LFSSM Reagent water containing 1000 mg/L LSSM. Described in Section 3.11 and3.13.
                                        314.1-30

-------
                                                                          EPA815-R-05-009
TABLE 5.  INITIAL DEMONSTRATION OF CAPABILITY QUALITY CONTROL
           REQUIREMENTS
   Method
  Reference
   Requirement
    Specification and
       Frequency
    Acceptance Criteria
  Section
  9.2.1
Demonstration of
Low System
Background
Analyze a LRB and
LFSSMB prior to any other
IDC steps.
Demonstrate that perchlorate is
below 1/3 of the MRL and that
possible interferences from
sampling protocols do not
prevent the identification and
quantification of perchlorate.
  Section
  9.2.4
Minimum
Reporting Limit
(MRL)
Confirmation
Fortify and analyze 7
replicate LFBs at the
proposed MRL
concentration. Calculate
the mean and the Half
Range (FIR). Confirm that
the Upper PIR and Lower
PIR (Sect.  9.2.4) meet the
recovery criteria.	
Section 9.2.4.2

Upper PIR < 150%.

Lower PIR > 50%.
  Section
  9.2.2
Demonstration of
Precision
Analyze 7 replicate LFBs
fortified near the mid-point
of the calibration curve
%RSDmustbe < 20%.
  Section
  9.2.3
Demonstration of
Accuracy)
Calculate average recovery
for replicates used in
Section 9.2.3.
Mean recovery + 25% of true
value.
  Section
  9.2.5
Validation of MRL
in 1000 mg/L
LFSSM
Analyze 7 replicate
LFSSMs fortified at the
MRL.
Section 9.2.4.2
                                                          Upper PIR < 150%.
                                                          Lower PIR > 50%.
  Section
  9.4.1
Quality Control
Sample
During IDC, each time a
new analyte PDS is made,
every time the instrument
is calibrated and at least
quarterly.	
The result for perchlorate must
be 75-125% of the true value.
                                          314.1-31

-------
                                                                           EPA815-R-05-009
TABLE 6.   ONGOING QUALITY CONTROL REQUIREMENTS (SUMMARY)
   Method
  Reference
   Requirement
     Specification and
        Frequency
     Acceptance Criteria
  Section
  10.2
Initial Calibration
Use external standard
calibration technique to
generate a first or second
order calibration curve. Use
at least 5 standard
concentrations.
Check the calibration curve
as described in Section 10.2.
                                 Analyze a QCS near the mid-
                                 point of the calibration
                                 curve.
When each calibration standard
is calculated as an unknown
using the calibration curve, the
result should be:
                                                                Level
                                                                  Result
                                                < MRL   + 50% True value

                                                > MRL     + 25% True value
                                                to high CAL

                                                The result for perchlorate must
                                                be 75-125% of the true value.

                                                Recalibration is recommended
                                                if these criteria are not met.
  Section
  9.3.2
Laboratory
Synthetic Sample
Matrix Blank
(LSSMB)
Daily, or with each Analysis
Batch of up to 20 field
samples, whichever is more
frequent.
Demonstrate that the
perchlorate is below V3 the
MRL, and confirm that possible
interferences do not prevent
quantification of perchlorate. If
the target exceeds V3 the MRL,
the results for perchlorate in the
Analysis Batch are invalid.
  Section
  9.3.3
Continuing
Calibration Check
(CCC) Standards
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.

Low CCC - at or below the
MRL concentration
Mid CCC - near midpoint in
calibration curve
High CCC - near the highest
calibration standard.
For each CCC the result must
be
                                                             CCC Level
                                                                  Result
                                                             < MRL   + 50% True value

                                                              > MRL     + 25% True value
                                                             to high CAL
                                           314.1-32

-------
                                                                          EPA815-R-05-009
TABLE 6. (Continued)
   Method
  Reference
   Requirement
     Specification and
        Frequency
     Acceptance Criteria
  Section
  9.3.4
Laboratory
Fortified Synthetic
Sample Matrix
CCCs
(LFSSM CCC)
In order to monitor trapping
efficiency during an Analysis
Batch, the CCC standards,
prepared in the LFSSM
(Sect. 9.3.3) are also required
at the same frequency and
concentrations.
For each LFSSM CCC the
result must be

CCC Level        Result
< MRL      + 50% True value

 > MRL     + 25% True value
to high CAL
  Section
  9.3.6
Laboratory
Fortified Sample
Matrix (LFSM)
Analyze one LFSM per
Analysis Batch (20 field
samples or less). Fortify the
LFSM with perchlorate at a
concentration close to but
greater than the native
concentration (if known).
Calculate LFSM recoveries.
Recoveries for the LFSM must
be calculated (Sect.  9.3.6.3 ).
The result must be

LFSM Level       Result
< MRL       + 50% True value

 > MRL      + 25% True value
to high CAL
  Section
  9.3.7
Laboratory
Duplicate (LD) or
Laboratory
Fortified Sample
Matrix Duplicate
(LFSMD)
Analyze at least one LD or
LFSMD daily, or with each
Analysis Batch (20 samples
or less), whichever is more
frequent.
Precision must be calculated
(Sect. 9.3.7.2). The result must
be
 Level              Result
<2xMRL     <  50%RPD
                                                            2xMRL
                                                            to high CAL
                                                               < 25%RPD
  Section
  9.4.1
Quality Control
Sample (QCS)
During IDC, each time a new
analyte PDS is made, every
time the instrument is
calibrated and at least
quarterly.	
Results must be +  25% of the
expected value.
  Section
Sample Holding
Time
28 days when processed and
stored according to sections
8.1 and 8.2 with appropriate
preservation and storage.
Sample results are valid only if
samples are extracted within
sample holding time.
                                           314.1-33

-------
                                                              EPA815-R-05-009
TABLE 7. SAMPLE ANALYSIS BATCH WITH QC REQUIREMENTS
Injection
#
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
Sample
Description
Laboratory Synthetic Sample Matrix Blank (LSSMB)
Low-CCC at the MRL (0.5 ng/L)
Low-Laboratory Fortified Synthetic Sample Matrix CCC
(LFSSM CCC @ 0.5 ng/L)
Sample 1
Sample 2
Sample 2 - Laboratory Fortified Sample Matrix (LFSM)
Sample 2 - Laboratory Fortified Sample Matrix Duplicate
(LFSMD)
Sample 3
Sample 4
Sample 5
Sample 6
Sample 7
Sample 8
Sample 9
Sample 10
Mid-CCCat5.0ng/L
Mid-Laboratory Fortified Synthetic Sample Matrix CCC
(LFSSMCCC@5.0ng/L)
Sample 1 1
Sample 12
Acceptance
Criteria
< 1/3 MRL
0.25 to 0.75 ng/L
0.25 to 0.75 ng/L
sample analysis
sample analysis
Recovery of 75 - 125%
%RPD = + 25%
sample analysis
sample analysis
sample analysis
sample analysis
sample analysis
sample analysis
sample analysis
sample analysis
3.75-6.5 |ig/L
3.75-6.5 |ig/L
sample analysis
sample analysis
                          CONTINUED on NEXT PAGE
                                   314.1-34

-------
TABLE 7. (Continued)
                                                                     EPA815-R-05-009
Injection
#
20
21
22
23
24
25
26
27
28
29
Sample
Description
Sample 13
Sample 14
Sample 15
Sample 16
Sample 17
Sample 18
Sample 19
Sample 20
High-CCC at 10 ng/L
High-Laboratory Fortified Synthetic Sample Matrix CCC
(LFSSMCCC@ 10ng/L)
Acceptance
Criteria
sample analysis
sample analysis
sample analysis
sample analysis
sample analysis
sample analysis
sample analysis
sample analysis
7.5 - 12.5 ng/L
7.5 - 12.5 ng/L
                                        314.1-35

-------
                                                                    EPA815-R-05-009
                                     Figure 1
EPA METHOD 314.1 CHROMATOGRAM of 5.0 fj.g/L C1O4 and 300 ^g/L 4-C1BSA WITH
                       IONPAC AS16 AND AS20 COLUMNS
         AS16 CIO4- Recovery = 79.0%
          AS20 CIO4- Recovery = 94.0%
      -0.10
        0.0  2.0  4.0  6.0  8.0  10.0 12.0 14.0 16.0 18.0 20.0 22.0 24.0 26.0 28.0 30.0 32.0   35.0
                                      314.1-36

-------
                                                                      EPA815-R-05-009
s.a
2.5
2.0
1.5
1.0
-O.I
-O.i
                                   Figure 2
        1C CHROMATOGRAM of SURFACE and GROUND WATER
                      FORTIFIED WITH 1.0  u.g/L C1O4
         Chlorinated surface water 1.6 ug/L
          Native level = 0.60 ug/L CIO4-
         Chlorinated ground water 1.1 ug/L
         Native level = 
-------
                                                                          EPA815-R-05-009
                                         Figure 3
1C CHROMATOGRAM of 3.0 ng/L C1O4 in 50, 500 and 1000 mg/L LSSM WITH the IONPAC
                                     AS16 COLUMN
      4.0-
      3.0
      2.0'
           MS
               3.0 |jg/L CICV in 1000 mg/L LSSM
               peak area = 0.0611 uS*min
               3.0 |jg/L CIO4' in 500 mg/L LSSM
               peak area = 0.0660 uS*min
               3.0 |jg/L CIO4" in 50 mg/L LSSM
               peak area = 0.0666 uS*min
    -0.201 > ' ' |  > ' ' | ' <  ' | ' < '  | ' ' ' |  ' ' > | ' '  < | ' ' '  | 'T ' i '  ' ' | ' '  ' | < ' ' |  < ' ' | '  ' ' | '
        0.0  2.0   4.0  6.0  8.0  10.0 12.0  14.0  16.0  18.0 20.0  22.0  24.0 26.0 28.0  30.0
                                          314.1-38

-------