USEPA CONTRACT LABORATORY PROGRAM
               *>
           STATEMENT OF WORK









             DIOXIN ANALYSIS




                Multi-Media




             Multi-Concentration








                 SOW. 9/86




                 Rev. 8/87





        Form IFB Series: WA86-K357

-------
STATEMENT OF WORK
TABLE OF CONTENTS

PREFACE:

EXHIBIT A:
EXHIBIT B:
EXHIBIT C:
EXHIBIT D:
EXHIBIT E:
EXHIBIT F:








Specifications for Chain-of -Custody, Document Control
and Standard Operating Procedures 	
No. of
Pages
. . . ii
. . iii
A-l-5
B-l-20
C-l-3
D-l-35
E-l-25
F-l-8

-------
                                 SCOPE OF WORK
The purpose of this contract Is to provide EPA with chemical analytical
services using selected Ion monitoring (SIM) gas chromatography/mass
spectrometry/data system (GC/MS/DS) techniques for the analysis of 2,3,7,8-
Tetrachlorodlbenzo-p-dloxin (TCDD) In soil or sediment and water samples.  The
majority of these samples are from areas of suspected 2,3,7,8-TCDD contami-
nation.  The methods required in this contract are effective for a concen-
tration range of 1 to 1000 parts per billion for soil or sediment and 0.01 to
10 parts per billion for water.

The Contractor shall use safe handling procedures and generally accepted good
laboratory practices to prepare and analyze for the presence of 2,3,7,8-
tetrachlorodibenzo-p-dioxin (TCDD) in soil or sediment and water samples.

The data obtained XSrill be used by EPA to determine the existence and extent of
threats to the public and the environment posed by hazardous waste disposal
sites.  The data may be used in civil and/or criminal litigation, therefore the
strictest adherence of chain-of-custody protocol, document control, and quality
assurance procedures is required.

-------
                         SUMMARY OF DIOXIN SOW CHANGES
                 In Updating the 9/83 Version to the 9/86 Version


 The method changes  that have  been  made resulted  in a  large  number of changes
 throughout the text.   A summary of the changes  Is  given  below,  a  line by line
 listing of all changes has  not been made.

 1.   The method has  been extended  to include  water  samples by  adding the
     extraction and  concentration steps from  Method 613.

 2.   Spiking solutions are now mixed with 1.5 mL of acetone  before addition to
     samples and blanks. This was  required by the  inclusion of  water in the
     method.

 3.   The surrogate concentration has been changed and  it  is  now  used to
     monitor method  detection  limits.

 4.   The S/N ratioiT^bf the 13C12~TCDD and native TCDD  are no longer
     reportables. The S/N ratio of 37ClA-2,3,7,8-TCDD m/e 328 is  a
     reportable.

 5.   The "Detection  Limit" has been retitled  "Maximum  Possible Concentration"
     and redefined.  The formula remains the  same.

 6.   Concentration Calibration (CC) solution  5 has  been eliminated, reducing
     the number of CC  solutions to  four and reducing the  calibration range and
     working range of  the method to 1 to 1000 ug/kg for soil/sediment and
     0.01 to 10 ug/L for water. The working  range  includes  using  a smaller
     sample aliquot  for the  higher  concentration  samples  (Exhibit  D Section 1.1).

 7   13C12-1򕂇4-TCDD has  been added as a recovery standard; added to the
     extract just before analysis  to monitor performance  of  the analytical
     train.   It is present in the  CC solutions  at  60% of  the internal standard
     (  Ci2~2,3,7,8-TCDD) concentration,  an advisory recovery window of
     40-120% has been set for internal standard recovery.  An action window
     may be  set when sufficient data is avaiable.
 8.   The confirmatory period scan requirement and the DFTPP tune requirement
     have been dropped.

 9.   The forms have been redesigned.

10.   The requirements for an acceptable method blank have been modified.

11.   Two concentration calibration options have been provided to facilitate
     the inclusion of the recovery standard and use of existing stocks of CC.
     solutions.

12.   Numerous changes have been made for clarity and consistency with other
     CLP methods.
                                       iii

-------
       EXHIBIT A




SUMMARY OF REQUIREMENTS

-------
              Exhibit A - Summary of Requirements
The Contractor shall provide appropriate equipment and experienced
personnel to Identify and measure 2,3,7,8-tetrachlorodibenzo-p-dioxin
(TCDD) in soil/sediment and water samples.  These analyses require GC/MS/DS
instrumentation including the capability to acquire, store, and retrieve
selected-ion-monitoring data for six ions.  Required equipment and exper-
tise is specified in IFB Pre-Award Bid Confirmations.  Samples to be
analyzed may contain high levels of toxic or hazardous materials and must
be stored and handled with appropriate precautions.

Specific analytical procedures to be used are provided in Exhibit D.
Specific QA/QC Requirements are specified in Exhibit E.  These procedures
must be followed explicitly without deviation, except as authorized in
writing by the Contracting Officer.  Required activities include:

1.1  Sample receipt and handling under Chain-of-Custody procedures.

1.1.1  Adherence to Chain-of-Custody procedures described in Exhibit G.
       Documentation described therein shall be required to show that
       all procedures are being strictly followed.  This documentation
       shall be reported as the complete Case File Purge.

1.2  Extract samples, perform column chroraatography procedures, and con-
     centrate extracts.  Analyze extract aliquots with GC/MS procedures.
     Required equipment:  capillary column, low resolution MS, and
     selected ion-monitoring data acquisition software.  (See Exhibit D.)

1.3  Acquire appropriate selected-ion-current profiles from MS data files.
     (See Exhibit D.)

1.4  Using criteria specified in Exhibit D, determine the presence or
     absence of 2,3,7,8-TCDD in each sample.  If present, calculate its
     concentration; if absent, calculate the maximum possible
     concentration.

1.5  For each sample extract or blank, measure signal to noise ratio of
     the surrogate compound,   Cl^-2,3,7,8-TCDD, and determine if the
     analyte detectability requirement has been met.  (See Exhibit E.)

1.6  For each sample extract or blank calculate the percent recovery of
     the internal standard (  C^"2,3,7,8-TCDD) using the recovery
     standard (L3C12-1,2,3,A-TCDD).

1.7  Periodically analyze performance check solution and appropriate QC
     samples (blanks, duplicates, fortified samples, and performance
     evaluation samples), as specified in Exhibit E.

1.8  Perform all required sample rerun extractions and analyses, as
     specified in Exhibit C.
                                A -  1

-------
     1.9   Provide all reports  and documentation within the applicable delivery
          requirement,  as  specified In Exhibit B.

     1.10 After receipt of samples, retain unused  portions of  samples and
          sample extracts  for  six months as specified In Exhibit  B.

2.  The Contractor shall receive  field samples In  groups designated  as sample
    "batches."   There will be  up  to twenty-four (24)  samples In a batch,  as
    defined below.

    2.1  Sample:  A sample Is  defined as a solid material (soil or sediment)
         or a liquid material  (water or rlnsate) that Is shipped  to  the
         Contractor for detection and measurement  of  2,3,7,8-tetrachloro-
         dlbenzo-p-dloxln  (TCDD).

    2.2  Field  Blanks (described  in Exhibit E, Sections 4.2 and 4.2.2):  One  or
         more field blank  samples will be included In each sample batch.

         2.2.1   Field Blank for Spiking  One field  blank for each  matrix
                will be designated for spiking. The  Contractor shall fortify
                this field blank  with 2,3,7,8-TCDD at a concentration of  1
                ug/kg for  soil/sediment or 10 ng/L for water.   After spiking,
                this field blank  shall be analyzed by SIM GC/MS;  it  shall not
                be analyzed prior to spiking.   If  a sample Is  not designated
                for spiking, the  laboratory shall  call SMO immediately.

         2.2.2   Unspiked Field Blanks  Other field  blanks, if any  are
                included in the batch, shall be extracted and  analyzed as
                routine samples per contract requirements.

    2.3  Rinsate Sample (described in Exhibit E, Section 4.2.3):   Normally, one
         rinsate sample will be included in each batch of samples, though more
         than one may be included.  The rinsate sample is a portion  of organic
         solvent that was  used to rinse sampling equipment, and therefore, will
         be liquid in nature,  rather than soil/sediment.  The  rinsate sample  Is
         to be prepared and analyzed as a routine  sample per contract require-
         ments.

3.  Extraction/Analysis Requirements:  Each sample in a batch shall  be
    extracted and analyzed with selected ion-monitoring (SIM)  GC/MS  procedures.
    Additionally, for each batch  of samples, three other analyses are required:

    3.1  Laboratory Method Blank  (described in Exhibit E, Section 4.1):  The
        Contractor shall extract  and analyze a laboratory method blank for
        each matrix in each batch of samples (or each time a group of samples
        are extracted)  using contract梥pecif led extraction and analysis
        procedures.  NOTE: Method blank analysis is considered part of the
        required Internal  laboratory QA/QC (included in the sample unit price);
        method blank analysis  Is  not considered as a separate sample analysis
        for contract accounting and/or billing purposes.

                                     A - 2

-------
    3.2  Duplicate Sample Analysts  (described  In Exhibit  E,  Section 5):   One
         sample of each matrix In each batch will be  designated  for duplicate
         analysis.  (In the event that no sample In the batch Is marked  for
         duplicate analysis,  then the Contractor shall  select sample(s)  from
         the batch and perform duplicate analysis.  DO  NOT USE THE FIELD BLANK
         FOR DUPLICATE ANALYSIS.)  A duplicate aliquot  of this sample shall be
         extracted and analyzed using contract-specified  extraction and  analysis
         procedures.   NOTE:  Duplicate sample  analysis  is accountable and
         blllable as  a separate sample analysis.

   3.3   Fortified Field Blank Analysis:   One  sample  for  each matrix In  each
         batch should be identified as a field blank  for  spiking (see Exhibit A
         section 2.2.1).  If no saraple(s) are  so designated the  laboratory
         should contact SMO immediately.


A.  Automatic Rerun Analyses (described in Exhibit C):  Certain  samples  may
    require sample reruns (reextraction and/or reanalysis) either due to
    problems with the sample matrix or Contractor insufficiencies.   NOTE:  Sam-
    ple reruns may be considered either as billable or  non-billable under the
    terms of this contract, as defined in Exhibit C.  For the purposes of this
    contract, the term "automatic rerun" shall signify  only billable rerun
    analyses.

5.  Summary of Batch  Analyses:

    5.1  Extractions  

               2A or  less field samples (including field  blanks  spiked at 1
                ug/kg or 10 ng/L as appropriate)
               1 or more laboratory method blanks*
               1 or 2 duplicate samples
           Plus, undetermined number of rerun  samples.**


    5.2  Analyses 

               24 or less field samples (including field  blanks
                spiked at 1 ug/kg or lOng/L as appropriate)
               1 or more laboratory method blanks*
               1 or 2 duplicate samples
           Plus, undetermined number of rerun  samples.**
   *Not billable as separate sample extraction/analysis (see Section 3.1)
  **A11 sample reruns may not be blllable as separate sample
    extraction/analyses, depending on basis for rerun (see Section A).
                                     A - 3

-------
6.  Contract Bid Lot:   One bid lot consists of analysis  of  a  maximum  number
    of three thousand  (3,000)  samples,  which will  be  received and  analyzed
    In batches of less than or equal to twenty-four (24) field samples.   The
    contractor will be required to perform a maximum  number of 200 sample
    analyses per calendar month period.

7.  Sample shipments to the Contractor's facility  will be scheduled and
    coordinated by the EPA CLP Sample Management Office  (SMO).

    7.1  The Contractor shall  communicate with SMO personnel  by telephone as
         necessary throughout  the process of sample scheduling,  shipment,
         analysis and  data reporting, to ensure that  samples  are properly
         processed. This shall include immediately notifying SMO  personnel of
         any irregularities with samples or sample paperwork  received (noting
         discrepancies from verbal order placed by SMO), problems  encountered
         in sample analyses that will affect the data produced,  and laboratory
         conditions that impact on timeliness of analyses and data reporting.
         In particular,  the Contractor  shall notify SMO  personnel  in  advance
         regarding sample data that will be late and  shall  specify an estimated
         delivery date.

    7.2  Sample analyses will  be scheduled by groups  of  samples, each defined
         as a Case and identified by a  unique EPA  Case and  Batch number.  A
         Case signifies a group of samples collected  at  one site or geo-
         graphical area over a predetermined time  period, and will include  one
         or more field samples with associated blanks.   Samples may be shipped
         to the Contractor in  a single  shipment or multiple shipments over  a
         period of time, depending on the size of  the Case.   If a  Case consists
         of multiple shipments, each shipment is considered a Batch.

    7.3  Each sample received  by the Contractor should be labeled  with an EPA
         sample number,  and accompanied by a Dioxin Shipment  Record bearing the
         sample number and descriptive  information regarding  each  sample.  The
         Contractor shall complete and  sign the Dioxin Shipment Record,
         recording the date of sample receipt and  sample condition on receipt
         for each sample container.  The Contractor shall submit the  signed
         copy of each  Shipment Record to SMO within seven (7) calendar days
         following sample receipt (see  contract delivery schedule).  If  there
         are problems  either with the samples (e.g. ,  mixed  media,  containers
         broken or leaking) or paperwork (e.g., Shipment Record not with
         shipment, sample and  Shipment  Record numbers do not  correspond)  the
         Contractor shall immediately contact SMO  for resolution.

    7.4  The EPA Case  and sample numbers shall be  used by the Contractor In
         Identifying samples received under this contract both verbally  and
         in reports/correspondence.                                          :

    7.5  Samples will routinely be shipped to the  Contractor  through  an  over-
         night delivery service.  However, as necessary, the  Contractor  shall b
         responsible for any handling or processing required  for the  receipt.oJ
         sample shipments, including pick-up of samples  at  the nearest
       .  servicing airport, bus station or other  carrier service within  the

                                     A - 4

-------
     Contractor's geographical area.  The Contractor shall be available
     to receive sample shipments at any time the delivery service Is
     operating, Including Saturdays.

7.6  The Contractor shall accept all samples scheduled by SMO,  provided
     that the total number of samples received In any calendar month does
     not exceed the monthly limitation expressed In the contract.  Should
     the Contractor elect to accept additional samples, the Contractor
     shall remain bound by all contract requirements for analysis of those
     samples accepted.

7.7  The Contractor shall be required to return sample shipping containers
     (e.g.,  coolers) to the return addressee indicated on or within the
     container, within a period of fourteen (14) days following receipt of
     the sample shipment.  The government will pay reasonable costs for the
     return of sample shipping containers by ground carrier service.
                                 A - 5

-------
               EXHIBIT B




REPORTING REQUIREMENTS AND DELIVERABLES

-------
     Exhibit B  -  Reporting Requirements and Deliverables


The Contractor shall provide reports and other deliverables as specified
in the Contract Schedule.  These reports are described below.  All reports
shall be submitted in legible form or resubmission shall be required.
All reports and documentation required, including chromatograms, shall be
clearly labeled with the EPA Case number, Batch number and associated
sample/Dioxin Shipment Record number(s).  If documentation is submitted
without the required identification, as specified above, resubmission
shall be required.

The Contractor shall provide all reports and deliverables as described
below.  The Contract Reporting Schedule (Section 2) specifies the number
of copies required, delivery schedule and distribution of all required
deliverables.

1.1  Dioxin~Shipment Record:  Copy of SMO Dioxin Shipment Record with lab
     receipt information and original Contractor signature.

1.2  Sample Data Summary Package:  Hard copy analytical data and documenta-
     tion are required from the Sample Data Package (Section 1.3).

     1.2.1  Case Narrative

     1.2.2  Completed data reporting sheets consisting of Forms B-l, B-2,
            B-3, and B-4.  Original and rerun sample data shall be provided
            on Form B-l.

1.3  Sample Data Package:  Hard copy analytical data and documentation
     are required as described below.  NOTE:  This analytical protocol is
     designed for receipt and analysis of samples by batch.  Therefore, it
     is desired that sample data from samples in one batch be reported
     together, i.e., on the same reporting form.  However, contract
     accounting and billing are based on the sample unit.

     1.3.1  Case Narrative (laboratory cover letter) contains the Case
            number, Dioxin Shipment Record numbers, Contract number and
            detailed documentation of any Quality Control, sample, ship-
            ment and/or analytical problems encountered in a specific
            Case.  Also included should be documentation of any internal
            decision tree process used along with a summary of corrective
            actions taken.  The Case narrative must be signed in original
            signature by the Laboratory Manager or his designate.

     1.3.2  Copies of completed Dioxin Shipment Records for all samples
            reported in data package.

     1.3.3  Results of initial triplicate analyses of four (4) concentra-
            tion calibration solutions, including all Selected Ion Current
                                B - 1

-------
            Profiles  (SICPs),  Calculated Response  Factors,  plotted concen-
            tration calibration curves  (see Section 9.2.6.5.7, Exhibit D),
            and computer  generated  quantitation reports.

     1.3.4  Completed  data  reporting sheets (Forms B-l,  B-2,  B-3,  and B-4)
            with appropriate SICPs.   Data results  of levels less than 10
            ug/kg  or  100  ng/L  shall  be  reported to two (2)  significant
            figures;  results greater than 10 ug/kg or 100 ng/L shall be
            reported  to three  (3) significant figures.  Apply the rounding
            rules  found in  Section  7.2.2, "Handbook for  Analytical Quality
            Control in Water and Wastewater Laboratories,"  EPA-600/4-79-
            019.  Each SICP must include the following header information:
            date and  time of analysis;  instrument  ID; and sample ID, i.e.,
            EPA sample number,  calibration solution number  (CC1, CC2, CC3,
            or  CCA) or column  performance check solution (PC).  When
            samples are analyzed more than once, all sample data shall be
          - ^reported.  (Note:   Original and rerun  data must be submitted.)

     1.3.5  SICPs  generated during  each performance check solution analysis
            and each concentration calibration solution  analysis.

     1.3.6  A chronological list of  all analyses performed.  If more than
            one GC/MS  system is used, a chronological list  is required for
            each system.  The  list  must provide the Data System File Name,
            the EPA sample  number, and  (if appropriate)  the contractor
            laboratory sample  number for each sample, blank,  concentration
            calibration solution, and performance  check  solution.   This
            list shall specify  date  and time of beginning of  analysis.  All
            sample/blank  analyses performed during a 12-hour  period must
            be  accompanied  by  two performance check solution analyses, one
            preceding  and one  following sample/blank analyses.  If multiple
            shifts are used, the ending performance check sample analysis
            from one  12-hour period  may serve as the beginning analysis
            for the next  12-hour period.

1.4  Monthly Sample Status  Report:   The Monthly Sample Status Report shall
     provide the status of  all samples  the Contractor has received or has
     had in-house  during  the calendar month.  Required status information
     includes:   samples received, samples extracted, samples analyzed,
     samples rerun.  All  samples shall  be identified by  appropriate EPA
     Sample, Case  and  Batch/Shipment numbers.

1.5  Daily Sample  Status  Report:  In response to verbal  request from the
     Sample  Management Office  or the Project Officer, the Contractor must
     verbally provide sample status information on a same-day basis.
     Should  written confirmation be requested, the Contractor must send
     daily sample  status  information in a written  form that same day using
     first-class mail  service.   Required Daily Sample Status information
     shall include the items noted  for the Monthly Sample Status Report
     and, in addition, shall require information on sample analysis reports
     in progress and  analysis  reports submitted/mailed.
                                B-2

-------
1.6  GC/MS Tapes:   The  Contractor must  store all raw GC/MS data  (includ-
     ing samples,  blanks,  concentration calibration solutions, performance
     check, solutions, and  performance evaluation samples)  on magnetic tape
     in appropriate instrument manufacturer's format.  The Contractor
     shall maintain a written reference/logbook of  tape files to EPA
     sample number, calibration data, standards and blanks.  The reference
     must  include  EPA sample numbers identified by  Case numbers and
     batch numbers. This  reference/logbook shall accompany tapes when
     submitted.

     The Contractor shall  submit GC/MS  tapes and associated reference/
     logbook within 7 days following receipt of written request by the EPA
     Project Officer or Sample Management  Office.  The Contractor shall
     retain tapes  for at least 365 days after data  submission unless
     submission is requested during that time.

1.7  Extracts and  Unused Sample Volume:  Unused portions of samples and
     all sample extracts shall be retained by Contractor for a period of
     365 days after data is submitted.   Extracts shall be stored at A癈;
     unused portions of samples can be  sealed and stored at ambient
     temperature.   Extracts and unused  sample volume containers shall be
     labeled with  EPA sample number, Case  number and Batch number.  A
     logbook of stored  extracts and sample volume shall be maintained,
     listing EPA sample numbers and associated Case and Batch numbers.

     The Contractor shall  submit sample extracts and/or unused sample
     volume and associated logbook(s) within 7 days following receipt of
     written request by the EPA Project Officer or  Sample  Management
     Office.  The  Contractor shall retain extracts  and unused sample
     volume for at least 365 days after data submission unless submission
     is requested  during that time.

     NOTE:   The Contractor is responsible  for shipment of  these materials
     in accordance with applicable Department of Transportation regulations.
     Whenever the  Contractor disposes of such materials, the Contractor
     is responsible for disposition of  these materials in accordance with
     applicable environmental regulations.

1.8  Complete Case File Purge: (formerly called the Document Control and
     Chain-of-Custody Package)  The Complete Case File Purge package includes
     all laboratory records received or generated for a specific Case or
     sample batch, that have not been previously submitted to EPA as a
     deliverable.   These items include  but are not  limited to:  sample
     tags,  custody records,  sample tracking records, analysts logbook
     pages, bench  sheets,  chromatographic charts, computer printouts, raw
     data  summaries, instrument logbook pages,  correspondence, and the
     document inventory.  (See Exhibit  F.)

2.    The following table (2.1) reiterates  the contract reporting and
     deliverables  requirements specified in the Contract Schedule and
     specifies the distribution that is required for each deliverable.
     Recipients include the CLP Sample  Management Office,  EMStr/LV QA.

                                B - 3

-------
          Division, the appropriate Regional Technical Officer, and NEIC
          Contract Evidence Audit Team.   NOTE:  Specific recipient names and
          addresses are subject to change during the term of the contract.
          The Project Offier will notify the contractor in writing of such
          changes when they occur.

     2. 1  Contract Reporting Schedule

                        CONTRACT REPORTING SCHEDULE
ITEM
 NO.
REPORT
   t
COPIES
DELIVERY
SCHEDULE
  REPORT DISTRIBUTION
SMO EMSL/LV REGION  NEIC
  1  Dioxin Shipment  1
       Record
  2  Sample Data      1
     Summary Package

  3  Sample Data      3
     Package

  4   Monthly Sample  2
      Status Report
      Daily Sample
      Status Report
            1
  6   GC/MS Tapes    Lot
  7  Extracts & Unused Lot
     Sample Volume
  8   Complete Case   1
      File Purge     Pkg
                 7 Days After Validated
                 Time of Sample Receipt
                 (VTSR)

                 21 days after VTSR
                 21 days after VTSR
       5 days following end of
       each calendar month

       Verbal and/or written;
       upon request by PO or
       SMO; maximum frequency
       is daily

       Retain for 365 days after
       data submission or submit
       within 7 days after receipt
       of written request by PO or
                    As Directed
                                                As Directed
                                                       SMO
                  Retain for 365 days after      As Directed
                  data, submission or submit
                  within 7 days after receipt
                  of written request by PO or SMO

                 Submit 180 days after data
                 submission or within 7 days
                 after receipt of written
                 request by PO or SMO
NOTE: ALL RESULTS SHALL BE REPORTED TOTAL AND COMPLETE.
      items 2 and 3 is required-
                                               Concurrent delivery  of
                                     B - 4

-------
2.2  Addresses for Distribution
     SMQ

     CLP Sample Management Office
     P. 0. Box 818
     Alexandria, VA  22313
     For overnight deliveries,  use
       street address:
     300 N. Lee St., Suite 200
     Alexandria,  VA  22314
EMSL/LV

USEPA EMSL/LV QA Division
Box 15027
Las Vegas, NV  89114
 ATTN:  Data Audit Staff

For overnight deliveries, use
  street address:
944 E. Harmon Ave.
Executive Center, Rm. 226
Las Vegas, NV  89109
     Region        "~

     Following contract award and prior to Contractor's receipt of the first
     batch of samples, the Sample Management Office, acting on behalf of the
     Project  Officer,  will provide the Contractor with the list of Deputy Project
     Officers for the  10 EPA Regions.   SMO will provide the Contractor with
     updated  Regional  address/name lists as necessary throughout the period of
     the contract.
     NEIC

     NEIC Contract Evidence
     Audit Team (CEAT)
     12600 West Colfax,  Suite  C310
     Lakewood,  CO  80215
                                       B - 5

-------
Lab:
Case/Batch No:

Instrument ID:
                FORM B-1S.   TCDD SOIL DATA REPORT FORM

               	           Report  Date:	

                                       Column:
                                                                    Page 1 of 2
EPA
Sample No.

















Extr.
Date

















Wet wt



......













ug/kg
Meas.

















TCDD
MFC

















GC/MS Analysis
Date

















Time

















Surr.
S/N Ratio

















*
Z REC(IS)

















 MB  =
  N  =
  D  =
 PE  =
MPC  =
*Note:
Method Blank                              FB
Native TCDD Spike                         IS
Duplicate/Fortified Field Blank           RR
EMSL-LV Performance Evaluation Sample     RS
Maximum Possible Concentration            ND
Relative to 13C12~1,2,3,4-TCDD
Field Blank
Internal Standard
Rerun
Recovery Standard
Not Detected
                                      B - 7
                                                                            9/86

-------
Lab:
Case/Batch No:

Instrument ID:
               FORM B-1S.  TCDD SOIL DATA REPORT FORM

              	           Report Date:	

                                        Column:
                                                                    Page 2 of 2
  EPA
       Rel.
Response Ratios
Response (Area)
Sample
Number
















320/
322
















332/
334IS
















332/
334RS
















259
















320
















322
















328
















332IS
















334IS
















332RS
















334RS
















 MB  =  Method Blank                               FB
  N  =  Native TCDD Spike                          IS
  D  =  Duplicate/Fortified Field Blank            RR
 PE  =  EMSL-LV Performance Evaluation Sample      ND
MPC  =  Maximum Possible Concentration             RS
                                                   Field Blank
                                                   Internal Standard
                                                   Rerun
                                                   Not Detected
                                                   Recovery Standard
                                     B - 8
                                                                    9/86

-------
Lab:
Case/Batch No:

Instrument ID:
 FORM B-1W.  TCDD WATER DATA REPORT FORM

	          Report Date:	

                        Column:
                                                                    Page 1  of 2
EPA
Sample No.

















Extr.
Date

















volume



...













ng/L
Meas.

















TCDD
MPC

















GC/MS Analysis
Date

















Time

















Surr.
S/N Ratio

















*
Z REC(IS)

















 MB  =  Method Blank                              FB
  N  =  Native TCDD Spike                         IS
  D  =  Duplicate/Fortified Field Blank           RR
 PE  =  EMSL-LV Performance Evaluation Sample     RS
MPC  =  Maximum Possible Concentration            ND
*Note:  Relative to 13C12~1,2,3,4-TCDD
                                   Field Blank
                                   Internal Standard
                                   Rerun
                                   Recovery Standard
                                   Not Detected
                                     B - 9
                                                     9/86

-------
Lab:
Case/Batch No:

Instrument ID:
              FORM B-1W.  TCDD WATER DATA REPORT FORM

                                        Report Date:	

                                        Column:
                                                                    Page 2 of 2
  EPA
       Rel.
Response Ratios
Response (Area)
Sample
Number
















3 20/
322
















332/
334IS
















332/
334 RS
















259
















320
















322
















328
















332IS
















334IS
















332RS
















334RS














'

 MB  =  Method Blank                               FB
  N  =  Native TCDD Spike                          IS
  D  =  Duplicate/Fortified Field Blank            RR
 PE  =  EMSL-LV Performance Evaluation Sample      ND
MPC  =  Maximum Possible Concentration             RS
                                                   Field Blank
                                                   Internal Standard
                                                   Rerun
                                                   Not Detected
                                                   Recovery Standard
                                     B -  10
                                                                    9/86

-------
A.   TCDD REPORT FORM (Form B-l)

     This form is used for tabulating and reporting case results.

     Complete the header information at the top of the page including instru-
ment ID, laboratory name, case/batch number, report date, and column used.

     EPA sample number is tabulated along with date sample was extracted, and
weight (wet) extracted to the nearest tenth (0.1) of a gram or volume extracted
(water) to the nearest 10 railliliters.

     Calculate the concentration of 2,3,7,8-TCDD using the formula:

                      Ax  QlS

                    Als . RRFn . W

            Cx  =---2,3,7,8-TCDD concentration in ug/kg or ug/L

            Ax  =  the sura of integrated ion abundance detected for m/z 320
                   and 322

                =  the sum of integrated ion abundances detected for m/z 332
                   and 334 (characteristic ions of 13C12-2,3,7,8-TCDD the
                   internal standard).

           Qis  =  quantity (in ng) of 13C12~2,3,7,8-TCDD added to the sample
                   before extraction
RRFn  =  calculated
         relative to
                              mean response factor for unlabeled 2,3,7,8-TCDD
                               13C19-2,3,7,8-TCDD
             W  =  The weight (in g) of soil/sediment extracted or volume of
                   water extracted (in mL)

     Positive samples are quantitated with values >10.0 ug/kg or 100 ng/L
     recorded to three (3) significant figures and those values <10.0 ug/kg or
     100 ng/L reported to two (2) significant figures.

     For samples in which unlabeled 2,3,7,8-TCDD was not detected calculate
the estimated maximum possible concentration, which is the concentration
required to produce a signal with a peak height of 2.5 times the background  晻
signal height.

     Use the formula:                                                     .'  .

                 2.5  . Hv . Q,--                                           '晻'晻'
                        A.    -1.0
         MPC  =  	
                            . W                                          -'''.
     where:  MPC  =  maximum possible concentration of  unlabeled  2,3,7,8-TCi)[>
                     required  to produce  Hx.

                                     B  -  11

-------
       Hx  =  peak height for m/z 320 or 322 in the same group
              of >5 scans used to measure Ajs.

      His  =  peak height for the appropriate ion characteristic of the
              internal standard,  m/z 332 when 320 is used to determine Ax,
              and m/z 334 when 322 is used to determine Ax.

      Qls  =  quantity (in ng) of 13C12~23.7,8-TCDD added to the
              sample before extraction.

     RRFn  =  calculated mean response factor for unlabeled 2,3,7,8-TCDD
              relative to 13C12-2,3,7,8-TCDD.

        W  =   weight (in g) of wet soil/sediment sample or volume of water
              extracted (in mL).

Report GC/MS Instrument ID, the date and time the analysis was performed,
and the signa~l~"to noise ratio for the surrogate compound.
                                 B  -  12

-------
                                    FORM B-2

                          INITIAL CALIBRATION SUMMARY
                        Page 1 of 2
Laboratory:
Case/Batch No.:
CC Solution Alternattve:

Instrument ID:
                                                AREA

Date















Time














Sol.
ID
CC1
CC1
CC1
CC2
CC2
CC2
CC3
CC3
CC3

CCA
CCA
CCA


320















322















328






*


t /
t /
/
/ t
/ t

332IS















33AIS















332RS















33ARS














Solution ID Codes;

CC1  Concentration calibration solution #1
CC2 = Concentration calibration solution #2
CC3 * Concentration calibration solution #3
CCA = Concentration calibration solution #A
  * Not present in CC Solution
    Alternative One.
                                     B - 13
                               9/86

-------
Laboratory:
Case/Batch No.:
                                    FORM B-2

                          INITIAL CALIBRATION SUMMARY

                         	  CC Solution Alternative:

                                            Instrument ID:
                                                                    Page 2 of 2


Date














Time













Sol.
ID
CC1
CC1
CC1
CC2
CC2
CC2
CC3
CC3
CC3
CCA
CCA
CCA
Measured
RRFn













Mean
RRFn













Measured
RRFi













Mean
RRFj[













Solution ID Codes:

CC1 = Concentration calibration solution #1
CC2 = Concentration calibration solution 92
CC3 = Concentration calibration solution #3
CCA = Concentration calibration solution #A
%RSD:
 CC1=
 CC2=
 CC3=
 CCA=
      RRF
         n
                              Native Mean
                              of Means:
                                                    IS Mean
                                                    of Means:
                                     B -  14
                                                                           9/86

-------
B.   Initial Calibration Summary (Form B-2)

     Record all routine calibrations (PCS and CC1)  performed during Initial
calibration on form B-3.

     Complete all header information including laboratory,  case/batch number,
and instrument ID and EPA CC Solution Alternative.

     Date and time along with response for each Ion is  recorded for each cali-
bration solution.  The response factors are calculated  with the following
equations:

RRFn (native Response Factor)       RRF^ (internal  Standard Response Factor)


         AV  Qi                                              Af_ . Qr<;
          A   * Lo                                               LQ    L o
RRFn
Where:
         AIs  Qn~                                             Ar
                                                                 s
 AX  =  the sum of Integrated ton abundance of ra/z 320 and 322 for unlabeled
        2,3,7,8-TCDD

Ais  =  tne sum 癴 integrated ion abundancces of m/z 332 and m/z 334 for
        13C12-2,3,7,8-TCDD

Ars  =  te sura 癴 integrated ion abundance of m/z 332 and m/z 334 for
          C12-1,2,3,4-TCDD

 Qn  =  quantity of unlabeled 2,3,7,8-TCDD injected


Qis  =  quantity of 13C12-2,3,7,8-TCDD injected

Qrs  =  quantity of 13C12-1,2,3,4-TCDD

     Calculate the mean RRF and the percent relative standard deviation for the
triplicate runs of each calibration solution.
                   SD
          %RSD  =   x
                   X
                                      B -  15

-------
Where:
     SD  =
 VN (Xt - X)2

1=1  N - 1
     X  =  mean of each of the three Response Factors respectively

     From the 4 mean native response factors and 4 mean Internal standard
     response factors:  calculate the mean of means for each respective RRF's.
                                     B -  16

-------
Laboratory: 	

Case/Batch No.
           FORM B-3

 ROUTINE  CALIBRATION  SUMMARY

	    CC Solution Alternative:

                     Instrument  ID:
                   (PCS) PERFORMANCE CHECK SOL.
                                         (CC1)
                                 CON.  CALIB.  SOL.  #1
1
Date
Time
Response
259
320
322
328
332IS
33AIS
332RS
33ARS
Ratios
320/322
332/334IS
332/334RS

RRFn

RRFi

Z Valley
























-....




































































































































































                                     B -  17
                                                  9/86

-------
C.   Routine Calibration Summary  (Form B-3)

     Complete the header Information Including the laboratory,  Instrument ID
     Case/Batch number and EPA CC Solution Alternative.
     For each performance check solution analyzed complete the date and time
     of analysis,  the response for m/z 259,  320,  and 322 for unlabeled 2,3,7,8-
     TCDD.  328 for 37C14~2,3,7,8-TCDD, and 332 and 334 for 13C12-2,3,7,8-TCDD
     and 13C12-1,2,3,4-TCDD.

     Ion ratios for m/z 320/322,  ra/z 332/334 for 13C12~2,3,7,8-TCDD and m/z
     332/334 for 13C12-1,2,3,4-TCDD are to be calculated and recorded.

     Response factors are to  be calculated as In the Initial Calibration Summary
     (Section B).

     For calculation of valley percent see Section D, Section 9.2.6.1.

     For each Concentration Calibration Solution #1 used In Routine Calibration,
     complete all the above information.
                                     B - 18

-------
                                    FORM B-A
                            QUALITY CONTROL SUMMARY
Laboratory Name
Case/Batch No.
Instrument ID
Accuracy, Fortified?
Spike Field Blank:
Relative Difference (%),
Duplicate Analysis:  	
                                      SOIL
                                     WATER
EPA Sample Number:

EPA Sample Number:
Accuracy, Fortified/
Spike Field Blank:
Relative Difference (%),
Duplicate Analysis:  	
EPA Sample Number:
EPA Sample Number:
                                     B -  19
                              9/86

-------
D,   QC Summary

     Complete all the header Information.

     Report the sample number for the fortified field blank and the % accuracy
of the fortified/spike field blank by using the following equation:

                    amount measured
     % accuracy  =  	  x 100
                          1.0

     Record the sample used for duplicate and the Relative Percent Difference
which is calculated as follows:

             |Sl - S2|
     RPD  =  	  x 100

              Sl-~_?2
                 2

     Where:

     Sj and $2 represent sample and duplicate sample results.
                                      B  -  20

-------
        EXHIBIT C




SAMPLE RERUN REQUIREMENTS

-------
                  Exhibit C - Sample Rerun Requirements


1.    Scope and Application

     The Contractor shall be required to reextract  and reanalyze certain sam-
     ples or batches of samples  in a variety of  situations  that may occur in
     the process of contract performance.   (For  purposes  of this contract,  the
     term "rerun" shall indicate sample reextraction,  cleanup and reanalysis.)

     In situations where the rerun is required due  to  matrix effects,  inter-
     ferences or other problems  encountered because of difficult samples, the
     Government will pay the Contractor for the  reruns.   Such reruns shall  be
     billable and accountable under the specified contract  allotment of auto-
     matic reruns.

     In situations where the rerun is required due  to  Contractor materials,
     equipment or~lnstrumentation problems, or lack of contractor's adherence
     to specified contract procedures, the rerun shall not  be billable nor
     accountable under the terms of the contract.

     Contractor's failure to perform any of the  sample reruns specified herein,
     either billable or non-billable, shall be construed  as Contractor non-
     performance and may result  in termination of the  contract for default  by
     Contractor.

     NOTE:  The only circumstance that may require  more than one rerun per
            sample is a contaminated method blank.

2.    Required Sample Reruns

     2.1  Automatic sample reruns, billable as such under the contract.

          2.1.1   If the calculated unlabeled TCDD  amount was outside the upper
                  initial calibration range, the Contractor shall reextract
                  the sample using a smaller sample aliquot, and reanalyze  the
                  sample.  (See Section 12.1.2,  Exhibit D.)

          2.1.2   If the internal standard was not  found to be present with at
                  least 10/1 signal to noise ratio  at  mass 332 and 334, the
                  Contractor shall reextract and reanalyze the sample.
                  NOTE: This rerun is billable only if the Contractor can
                  demonstrate that the internal standard was added to the
                  sample in accordance with contract specifications.  (See
                  Sections 3.11 and 11.6.3, Exhibit D.)
                                     C -  1

-------
     2.1.3  If the internal standard 332/334 ratio was outside of the
            contract specified control limits of 0.67-0.90,  the Contractor
            shall reextract and reanalyze the sample.   (See  Section
            11.6.5, Exhibit D.)  This reanalysis is billable,  only if
            the internal standard 332/334 ratio is still outside the
            0.67-0.90 control limits.

     2.1.4  If the isotope abundance ratio for m/z 320/322 is  less than
            0.67 or greater than 0.90 and all other criteria contained in
            Section 11.6.1 through 11.6.5 of Exhibit D are met, then the
            sample must be rerun unless the MFC is 
-------
          2.2.5  If the accuracy of the  measured  concentration of  native
                 2,3,7,8-TCDD In the spiked  (fortified)  field blank Is  not
                 between 60-140%, the contractor  shall reextract and reanalyze
                 a second aliquot of the fortified  field blank sample.

3.    Sample Rerun QC

     3.1  A native spike and duplicate shall be performed for each batch of
          samples reanalyzed as specified in Section 2.

          3.1.1  If a concurrent Dioxin Case is being processed, the native
                 spike and duplicate from that  case may  be shared  with  the
                 rerun samples if the total  number  of samples does not  exceed
                 24.  If the total number of samples exceeds  24 an additional
                 native spike and duplicate  must  be analyzed  as in Section
                 3.1.2 below.  The native spike and duplicate data shall be
                 reported in each Case Data  Package.  (Note:   The  QC samples
                 are to be billed only under the  Case number  in which the QC
                 sample was received.)

          3.1.2  If no other Dioxin Case is  being processed at the time of
                 reanalysis, the native spike and duplicate shall  be chosen
                 from the Case and batch in  which the rerun sample belongs.
                 The QC samples are to be billed  only if the  rerun samples
                 are billable according to Section  2.
                                     C - 3

-------
    EXHIBIT D




ANALYTICAL METHODS

-------
                                  INDEX
Section
 Number                          Subject
   1.         Scope and Application
   2.         Summary of Method
   3.         Definitions
   4.         Interferences
   5.         Safety
   6.         Apparatus and Equipment
   7.         Reagents and Consumable Materials
   8.         Sample Preservation and Handling (Ref. Exhibit G)
   9.         Calibration
  10.         Quality Control (Ref. Exhibit E)
  11.         Analysis Procedures
  11.1        Soil Sample Extraction
  11.2        Water Sample Extraction
  11.3        Column Chromatography
  11.4        Carbopack Column Chromatography Procedure
  11.5        GC/MS Analysis
  11.6        Identification Criteria
  12.         Calculations
                                      D - 1

-------
                         Exhibit D - Analytical Methods

                  2,3,7,8-Tetrachlorodlbenzo-p-dloxln In Soil,
           Sediment and Water by High Resolution Gas Chromatography/
                        Low Resolution Mass Spectrometry


1.   SCOPE AND APPLICATION

     1.1  This method provides procedures for detection and measurement of
          2,3,7,8-tetrachlorodibenzo-p-dloxln (2,3,7,8-TCDD; CAS Registry Num-
          ber 1746-01-6;  STORET Number 34675) at concentrations of 1 ug/kg to
          100 ug/kg In 10-g allquots of wet soil/sediment and 10 ng/L to 1000
          ng/L in 1 L aliquots of water.  The use of 1 g aliquots permits
          measurement of concentration up to 1000 ug/kg in soil/sediment.  The
          use of a 100 mL aliquot volume permits measurement of concentrations
          up to 10_ug/L in water.

     1.2  CAUTION: The analysis of water samples includes whatever particulates
                   may be present.  The estimated solubility of 2,3,7,8-TCDD in
                   water is less than 50 ng/L^, therefore positive values above
                   this level should be considered to be a function of the TCDD
                   associated with the particulates rather than the water.

     1.3  The minimum measurable concentration is estimated to be 0.3 ug/kg or
          3 ng/L, but Is dependent on Interfering compounds present In the
          sample matrix.

     1.4  This method is designed for use by analysts who are experienced in
          the use of a gas chromatograph/mass spectrometer.

     1.5  CAUTION:  Because 2,3,7,8-TCDD is extremely toxic, safety procedures
                    described in Section 5 of this method should be followed to
                    prevent exposure of laboratory personnel to materials
                    containing this compound.


2.   SUMMARY OF METHOD

    2.1  Soil/Sediment Extraction;  For purposes of this contract a soil/sedi-
         ment sample is defined as a portion of wet soil or sediment which may
         contain  other solids such as stones, vegetation etc., but  should not
         contain  an obvious liquid phase (See Exhibit D, Section 8.3.2);  
         Fifty (50) ng of  13C12~labeled 2,3,7,8-TCDD and 1.4 ng of   C1A-
         labeled  2,3,7,8TCDD are added  to  a  10 g aliquot of wet soil or
         sediment sample,  the sample aliquot  Is mixed with  20  g of  anhydrous
         sodium sulfate and is extracted with a mixture of  hexane and  metharol
         by agitating the  sample aliquot and  solvent  continually  in a  glass,
         jar.
                                      D  -  2

-------
2.2  Water Extraction:  For the purpose of this contract a water sample Is
     defined as a single phase system that Is primarily water but may contain
     small amounts of floating, suspended, and settled partlculate matter.
     Multiple phases should not be present (See Exhibit D, Section 8.3.2.).
     Fifty (50) ng of 13C12-labeled 2,3,7,8-TCDD and 1.4 ng of 37Cl4-labeled
     2,3,7,8TCDD are added to approximately 1 L of water and extracted with
     methylene chloride using a separatory funnel.  The methylene chloride
     extract Is exchanged to hexane during concentration.

2.3  Cleanup and Analysis:  Column chromatographlc procedures are used to help
     eliminate sample components that may interfere with detection and measure-
     ment of 2,3,7,8-TCDD.  The extract Is concentrated to 50 uL, 50 ng
     13C12-1,2,3,4-TCDD are added, and a 2 uL aliquot is Injected into a fused
     silica capillary column in a gas chromatograph (GC) interfaced to a mass
     spectrometer (MS) that has at least unit resolution at m/z 334.  Identifi-
     cation of 2,3,7,8-TCDD is based on detection of three ions, measurement of
     the appropriate relative abundances of two characteristic ions in the
     molecular ion cluster, and determination of the retention time of the
     sample analyte relative to the internal standard,   C^2-2,3,7,8-TCDD,
     contained in the sample extract.  The 2,3,7,8-TCDD concentration is
     determined by measuring the MS response to the sample component relative
     to the MS response to  3C12-2,3,7,8-TCDD (the internal standard).  The
     labeled internal standard method presumes that internal standard losses
     during method procedures are equal to unlabeled TCDD losses.  Therefore,
     the calculated sample 2,3,7,8,-TCDD concentration Is corrected for losses
     during sample preparation.

     The   Cl^-2,3,7,8-TCDD is a surrogate compound that is added to each
     sample and is analyzed exactly the same as unlabeled TCDD.  The surrogate
     compound is used to determine that the detection criteria for unlabeled
     2,3,7,8-TCDD in the same sample have been met.

         13
     The   Cj2~l2,3,4-TCDD Is a recovery standard that is added to each sample
     and blank.  The recovery of the internal standard (13C12~2,3,7,8-TCDD) and
     the surrogate compound (37Cl^-2,3,7,8-TCDD) are related to the precision
     and sensitivity of the analysis for unlabeled 2,3,7,8-TCDD in the sample.

3.   DEFINITIONS

     3.1  Concentration calibration solution  a solution containing known
          amounts of the analyte (unlabeled 2,3,7,8-TCDD), the surrogate
          compound (37Cl^-2,3,7,8-TCDD), the recovery standard (13C12~1,2,3,4-
          TCDD) and the Internal standard (13C12~2,3,7,8-TCDD);  it  is used
          to determine the instrument response of the analyte compound  relative
          to the  Internal standard and  the recovery of the Internal standard.

     3.2  Field blank  a portion of soll/sediraent or water uncontamlnated.
          with 2,3,7,8-TCDD submitted with the samples.
                                      D -  3

-------
 3.3  Rlnsate   a portion of  trichloroethylene used to rinse sampling
      equipment and analyzed to demonstrate that samples were not contami-
      nated during sampling.

 3.4  Internal standard  13C12-2,3,7,8-TCDD,  which Is added to every
      sample and is present at  the same concentration In every blank,
      quality control sample, and concentration calibration solution.   It
      Is added to the samples before extraction and is used to measure the
      concentration of the analyte.

 3.5  Recovery standard    C12~~* ,2,3,4-TCDD is added to every extract and
      is present in all standards.  It  is added to every extract just
      before analysis and is used to measure the recovery of the internal
      standard.

                                                     37
 3.6  Surrogate compound  A known concentration of   Cl^-2,3,7,8-TCDD,
      which is added to all samples before analysis.  Its signal to noise
      ratio is measured in each sample, and is  used to indicate that
      unlabeled 2,3,7,8-TCDD is detectable at less than 1.0 ug/kg or
      10 ng/L.

 3.7  Laboratory method blank  a blank prepared in the laboratory by
      performing all analytical procedures except addition of a sample
      aliquot to the extraction vessel.

 3.8  Performance check mixture  a mixture of known amounts of selected
      standard compounds; it is used to demonstrate continued acceptable
      performance of the GC/MS/DS system.

 3.9  Performance evaluation sample   a soil/sediment or water sample
      containing a known amount of unlabeled 2,3,7,8-TCDD.  It is distri-
      buted by EPA to potential contractor laboratories who must analyze it
      and obtain acceptable results before being awarded a contract for
      sample analyses (see IFB Pre-Award Bid Confirmations).  It may also
      be included as an unspecified QC sample in any sample batch submitted
      to the laboratory for analysis.

3.10  Response factor  response of the mass spectrometer to a known
      amount of an analyte relative to a known amount of an internal
      standard.

3.11  Signal-to-noise (for the purpose of  this contract)  is defined as  the
      ratio of analyte signal to  random background  signal.  Display-each
      characteristic ion using a window 20 scans wide and centered abound
      the elution time of 2,3,7,8-TCDD.  Draw a base line from the lowest
      point  in the 20 scan window.  The noise  is defined  as the hei|-,h'C.  of
      the largest signal  (excluding signal due to TCDD or other chemicals)
      on either  side of  the 2,3,7,8-TCDD peak, within  the 20  scan wiuJcw.
      The signal  is defined as the height of 2,3,7,8-TCDD peak.  Chemical
      noise  is left to the judgement of  the analyst.
                                 D -

-------
     3.12  Abbreviations

          TCDD - Tetrachlorodibenzo-p-dloxln
          GC   - Gas Chromatograph
          MS   - Mass Spectrometer
          DS   - Data System
          SIM  - Selected Ion Monitoring
          SMO  - Sample Management Office
          SICP - Selected Ion Current Profile
          S/N  - Signal to Noise
          PC   - Performance Check Standard
          CC   - Calibration Check Standard
          RRF  - Relative Response Factor
          IS   - Internal Standard
          RS   - Recovery Standard
          K-D   Kuderna-Danish Apparatus
          OD   - Outside Diameter
          m/z  - Mass to Charge Ratio
          MPC  - Maximum Possible Concentration
          VTSR - Verified Time of Sample Receipt

4.   INTERFERENCES

     Any compound that yields ions at m/z 259, 320, 322, or 328 and also
     elutes within 10 scans of the internal standard is a potential interfer-
     ence.  Most frequently encountered interferences are other sample compo-
     nents that are extracted along with TCDD.  Because very low levels of TCDD
     must be measured, elimination of interference is essential.  High purity
     reagents and solvents must be used and all equipment must be scrupulously
     cleaned.  Laboratory method blanks (Exhibit E, Quality Control, Section 4)
     must be analyzed to demonstrate lack of contamination that would interfere
     with TCDD measurement.  Column chromatographic procedures are used to
     remove some coextracted sample components; these procedures must be per-
     formed carefully to minimize loss of TCDD during attempts to enrich its
     concentration relative to other sample components.

5.   SAFETY

     5.1  The toxicity or carcinogenicity of each reagent used in this method
          has not been precisely defined; therefore, each chemical compound
          should be treated as a potential health hazard.  From this viewpoint,
          exposure to these chemicals must be reduced to the lowest possible
          level by whatever means available.  The laboratory is responsible "tor
          maintaining a file of current OSHA regulations regarding the safe
          handling of the chemicals specified in this method.  A reference file
          of material data handling sheets should also be made available to all
          personnel involved in the chemical analysis.  Additional references
          to laboratory safety are identified.C1-3)  2,3,7,8-TCDD has been
          identified as a suspected human or mammalian carcinogen.
                                     D - 5

-------
5.2  Each laboratory must develop a strict safety program for handling
     2,3,7,8-TCDD.   The following laboratory practices are recommended:

     5.2.1  Contamination of the laboratory will be minimized by con-
            ducting all manipulations in a hood.

     5.2.2  The effluents of sample splitters for the gas chromatograph
            and roughing pumps on the GC/MS should pass through either a
            column of activated charcoal or through a trap containing oil
            or highboiling alcohols.

5.3  The following precautions for safe handling of 2,3,7,8-TCDD in the
     laboratory are presented as guidelines only, and are based on safe
     handling practices included in USEPA Method 613.(*)  The precautions
     for safe handling and use are necessarily general in nature because
     detailed, specific recommendations can be made only for the par-
     ticular_exposure and circumstances of each individual usage.
     Assistance in evaluating the health hazards of particular laboratory
     conditions may be obtained from certain consulting laboratories and
     from State Departments of Health or of Labor, many of which have an
     industrial health service.  Although 2,3,7,8TCDD is extremely toxic
     to laboratory animals, it has been handled for years without injury
     in analytical and biological laboratories.  Techniques used in
     handling radioactive and infectious materials are applicable to
     2,3,7,8-TCDD.

     5.3.1  Protective Equipment:  Throw-away plastic gloves, apron or lab
            coat, safety glasses and lab hood adequate for radioactive
            work.

     5.3.2  Training:  Workers must be trained in the proper method of
            removing of contaminated gloves and clothing without
            contacting the exterior surfaces.

     5.3.3  Personal Hygiene:  Thorough washing of hands and forearms
            after each manipulation and before breaks (coffee, lunch, and
            shift) with any mild soap and plenty of scrubbing action.

     5.3.A  Confinement:  Isolated work area, posted with signs;
            segregated glassware and tools; and plastic-backed absorbent
            paper on benchtops.

     5.3.5  Waste:  Good technique includes minimizing contaminated waste.
            Plastic bag liners should be used in waste cans.  Janitors
            should not handle wastes.

     5.3.6  Disposal of Wastes:  2,3,7,8-TCDD decomposes above 800.C.  .Low
            level waste, such as the absorbent paper and plastic glovcc,
            may be burned in a good  incinerator.  Waste containing gtoss
            quantities (milligrams) of 2,3,7,8-TCDD should be packaged
            securely and disposed  through  commercial or governmental
                                D - 6

-------
        channels that are capable of  handling high-level or extremely
        toxic wastes.  Liquids should be allowed to evaporate In a
        good hood and In a disposable container; residues may then be
        handled as above.

 5.3.7  Glassware, Tools,  and Surfaces:   Satisfactory cleaning may be
        accomplished by rinsing with  1,1,1-trichloroethane, then
        washing with any detergent and water.  Dishwater may be
        disposed to the sewer.  (Also see Section 6.5.)

 5.3.8  Laundry:  Clothing known to be contaminated should be disposed
        with the precautions described under Section 5.3.6.  Lab coats
        or other clothing worn In 2,3,7,8-TCDD work may be laundered.
        Clothing should be collected  in  plastic bags.  Persons who
        convey the bags and launder the  clothing should be advised of
        the hazard and trained In proper handling.   The clothing may
        be put into a washer without  contact if the launderer knows
        the problem.  The washer should  be run through a cycle before
        being used again for other clothing.  Disposable garments may
        be used to avoid a laundry problem,  but they must be properly
        disposed or incinerated.

 5.3.9  Wipe Tests:  A useful method  to  determine cleanliness of work
        surfaces and tools is to wipe the surface with a piece of
        filter paper, which is extracted and analyzed by gas chromato-
        graphy (limit of sensitivity  of  approximately 0.1 ug per
        wipe).  Less than A pg/cm2 2,3,7,8-TCDD indicates acceptable
        cleanliness; anything higher  warrants further cleaning.  More
        than AGO pg/cm^ indicates an  acute hazard that requires prompt
        cleaning before further use of the equipment or work space and
        indicates that unacceptable work practices have been employed
        in the past.

5.3.10  Inhalation:  Any procedure that  may produce airborne contami-
        nation should be performed with  good ventilation.  Gross
        losses to a ventilation system should not be allowed.  Han-
        dling of the dilute solutions normally used in analytical and
        animal work presents no inhalation hazards except  in case of
        an accident.  Finely divided  soils contaminated with 2,3,7,8-
        TCDD are hazardous because of the potential for inhalation.
        Such samples should be handled in a confined environment, such
        as a hood or glove box, or laboratory personnel should wear
        masks fitted with a particulate  filter and charcoal sorbent.

5.3.11  Accidents:  Remove contaminated  clothing immediately, taking
        precautions not to contaminate skin or other articles.  Wai,h
        exposed skin vigorously and repeatedly until medical attention
        is obtained.
                            D - 7

-------
6.  APPARATUS AND EQUIPMENT

    6.1  Gas Chromatograph/Mass Spectrometer/Data System (GC/MS/DS)

          6.1.1  The GC must be capable of temperature programming and be
                 equipped with all required accessories, such as syringes,
                 gases, and a capillary column.  The GC Injection port must be
                 designed for capillary columns.  Splltless or on-column Injec-
                 tion technique Is recommended.  With this method, a 2 uL
                 Injection volume Is used consistently.  However, with some GC
                 Injection ports other volumes may be more appropriate.   Any
                 volume that produces adequate precision sensitivity, and
                 chromatographlc separation may be used.  A 1 uL Injection
                 volume may be used If adequate sensitivity and precision can
                 be achieved,  CAUTION:  The injection volume for all extracts,
                 blanks, calibration solutions and the performance check samples
                 must be the same.

          6.1.2  Mass spectral data are obtained with electron ionization at a
                 nominal electron energy of 70 eV.  To ensure sufficient pre-
                 cision of mass spectral data, the required MS scan rate must
                 allow acquisition of at least five data points for each of six
                 ions while a sample component elutes from the GC.

          6.1.3  An interfaced data system (DS) is required to acquire,  store,
                 reduce and output mass spectral data.  The DS must be equipped
                 with a selected ion monitoring (SIM) program to acquire data
                 for at least six ions that are characteristic of labeled and
                 unlabeled 2,3,7,8-TCDD.  (The mass spectrum of unlabeled
                 2,3,7,8-TCDD is shown in Figure 1 at the end of this Exhibit.)
                 The same integration time must be used for each ion monitored,
                 and the integration time used for sample analyses must be the
                 same as the time used to analyze concentration calibration
                 solutions and the performance check solution.  Total data
                 acquisition time per cycle (six ions) must not exceed 1.5
                 seconds.

          6.1.4  The Contractor shall use a magnetic media storage device
                 capable of recording data suitable for long-term off-line
                 storage.  The Contractor shall record all raw GC/MS data
                 acquired during the entire contract period on magnetic media
                 in appropriate instrument manufacturer format.  The Contractor
                 shall provide the data on 9-track magnetic tape in appropriate
                 instrument manufacturers format to the US EPA within sever. (7)
                 days of request by the Project Office or SMO.  The tape=> iiuist
                 be retained by the contractor  for 180 days after data package
                 submission unless requested by EPA.

    6.2  GC Column  Two fused silica capillary columns  are  recommended; cue
         is a 60-m SP-2330 and the other  is a  50-m CP-SIL 88.   Any capillary
         column  that separates 2,3,7,8-TCDD from all  other TCDDs may be  uee-i,
         but this separation must be demonstrated.  Minimum acceptance criteria
         must be determined per Section 9.2.6.1.  At  the  beginning-and er.u  of

                                     D -  8

-------
     each 12-hour period during which sample or concentration calibration
     solutions will be analyzed, column operating conditions must be
     demonstrated to achieve the required separation on the column to be
     used for samples.  Operating conditions known to produce acceptable
     results with the recommended columns are shown in Table 1 at the end
     of this Exhibit.  It is the Contractor's responsibility to verify
     whether the information in Table 1 is suitable for the laboratory's
     instrument(s).

6.3  Miscellaneous Equipment

     6.3.1  Nitrogen evaporation apparatus with variable flow rate.

     6.3.2  Mechanical shaker  A magnetic stirrer or a wrist-action or
            platform-type shaker that produces vigorous agitation.

     6.3.3  Analytical balance capable of accurately weighing 0.01 g.

     6.3.4  Centrifuge capable of operating at 400 x G.

     6.3.5  Water bath  equipped  with concentric ring cover and
            temperature controlled within ^2癈.

     6.3.6  Stainless steel spatulas or spoons.

     6.3.7  Stainless steel (or glass) pan large enough to hold contents
            of 1-pint sample containers.

     6.3.8  Glove box.

6.4  Glassware

     6.4.1  Extraction jars  amber glass with Teflon-lined screw cap;
            minimum capacity of approximately 500 mL; must be compatible
            with mechanical shaker to be used.

     6.4.2  Kuderna-Danish apparatus  500-raL evaporating flask, 10-mL
            graduated concentrator tubes with ground-glass stoppers,
            3-ball macro-Snyder column, and 2-ball micro-Snyder column.

     6.4.3  Culture tubes  8-mL glass.

     6.4.4  Mini-vials  1-mL  amber borosilicate glass with coniccl-
            shaped reservoir and screw caps lined with Teflon-faced
            silicone  disks.

     6.4.5  Funnels  glass; appropriate  size  to accommodate  filter paper
            used to filter  jar  extract  (volume  of approximately 170  .nL;.

     6.4.6  Chromatography  columns   1  cm ID x 20 cm long and 1  cm  II)
            x  30 cm  long.
                              D  -  9

-------
          6.4.7  Separatory funnels  2 L with Teflon stopcock.

          6.4.8  Drying column 19 mm ID glass chromatographlc column with a
                 coarse frit (a small pad of pyrex glass wool may be
                 substituted for the frit to avoid cross contamination).

          6.4.9  Boiling chips  Approximately 10/40 mesh.  Silicon carbide
                 or Teflon may be used.  Heat to 400癈 for 30 mln or Soxhlet
                 extract with methylene chloride as appropriate.

     6.5  NOTE:  Reuse of glassware should be minimized to avoid the risk of
                 using contaminated glassware.  All glassware that Is reused
                 must be scrupulously cleaned as soon as possible after use,
                 applying the following procedure.  Rinse glassware with the
                 last solvent used In It.  Wash with hot water containing
                 detergent.  Rinse with copious amounts of tap water and
                 several portions of distilled water.  Drain dry and heat In a
                 muffle furnace at 400癈 for 15 to 30 rain.  Volumetric glass-
                 ware should not be heated in a muffle furnace, and some
                 thermally stable materials (such as PCBs) may not be removed
                 by heating in a muffle furnace.  In these cases, rinsing with
                 high-purity acetone and hexane may be substituted for muffle
                 furnace heating.  After glassware is dry and cool, store
                 inverted or capped with aluminum foil in a clean environment.

       CAUTION:  The analysis for 2,3,7,8-TCDD in water is for much lower con-
                 centrations than In soil/sediment.  Extreme care must be
                 taken to prevent cross梒ontamination between soil and water
                 samples.  It is strongly recommended that separate glassware
                 be reserved for analyzing water samples.  It is recommended
                 that all glassware be rinsed with solvent immediately before
                 use and that the pooled solvent for a set of extractions be
                 concentrated and analyzed as a method of demonstrating that
                 the glassware was free of contamination.

7.  REAGENTS AND CONSUMABLE MATERIALS

    7.1  Column Chromatography Reagents

          7.1.1  Alumina, acidic AG4, Bio Rad Laboratories (catalog #132-1240
                 or equivalent)  Soxhlet extract with methylene chloride for
                 21 hours and activate by heating in a foil covered glass
                 container for 24 hours at 190癈.

          7.1.2  Silica gel  high purity grade, type 60, 70-230 mesh; Goxhlet
                 extract with methylene chloride for 21 hours and activate by
                 heating In a foil-covered glass container for 24 hours at
                 130癈.

          7.1.3  Silica gel Impregnated with sodium hydroxide  Add one part
                 of 1 M NaOH solution to two parts of silica gel (extracted and
                 activated) In a screw梒ap bottle and mix with a glass  rod
                 until free of lumps.

                                     D - 10

-------
     7.1.4  Silica gel Impregnated with 40% (by weight) sulfurlc acid 
            Add two parts (by weight) concentrated sulfurlc acid to three
            parts (by weight) silica gel (extracted and activated), mix
            with a glass rod until free of lumps, and store In a screw-
            capped glass bottle.

     7.1.5  Sulfurlc acid,  concentrated  ACS grade, specific gravity
            1.84.

     7.1.6  Graphitized carbon black (Carbopack C or equivalent), surface
            area of approximately 12 m2/g, 80/100 mesh.

     7.1.7  Celite 545, reagent grade,  or equivalent.

7.2  Filter paper  Whatman No.  1 or equivalent; rinse with hexane
     before use.

7.3  Glass wool, silanized  Extract with methylene chloride and then
     hexane before use.

7.4  Sodium sulfate  Granular,  anhydrous; before use, extract with
     methylene chloride and dry for >4 h in a shallow tray placed in an
     oven operated at 120癈.

7.5  Solvents  High purity, distilled-in-glass; hexane, methanol,
     methylene chloride, toluene, and isooctane.

7.6  Concentration Calibration Solutions (Table 2)  EMSL-LV will provide
     the concentration calibration solutions in either of two formulations
     depending on the availability of standard materials.  Alternative one
     (Section 7.6.1) includes the   C12-1,2,3,4-TCDD recovery standard in
     the concentration calibration solutions.  Alternative two (Section
     7.6.2 requires the addition of a specified amount of   C^2-l,2,3,4-
     TCDD to 1 mL of each concentration calibration solution.  The
     solutions obtained will be clearly labeled to identify which
     formulation Is supplied.

     7.6.1  Alternative One

                                                               ,8-TCDD at
                                                               internal
                                                               (the
            recovery standard) at a constant concentration.  Two of these
            solutions also contain  7Cl^-2,3,7,8-TCDD  (the surrogate com-
            pound, CASRN 85508-50-5) at a constant concentration.  ' .Concen-
            tration calibration solutions are  to be obtained from  the
            Quality Assurance Division, US EPA  Environmental Monitocittft
            Systems Laboratory (EMSL-LV), Las Vegas, Nevada.  However, If
            not  available from EMSL-LV, standards may  be  obtained  from
            commercial  sources, and solutions may be prepared in the
            contractor  laboratory.  Traceability of  standards must te
                                D - 11

-------
       verified against EPA-supplled standard solutions, by labora-
       tory SOP's as required In IFB Pre-Award Bid Confirmations,
       part 2.f.(4).

       7.6.1.1  Each of solutions #l-#4 contains 13C12~2,3,7,8-
                TCDD at a concentration of 1 ng/uL which Is
                equivalent to a 50-uL extract of a 10-g sample to
                which that compound (the Internal standard) was
                added at a concentration of 5 ug/kg or a 50 uL
                extract of a 1 L water sample to which the
                Internal standard was added at a concentration of
                50 ng/L.

       7.6.1.2  Solutions #1-M contain unlabeled 2,3,7,8-TCDD
                at concentrations of 0.2, 1, 5, and 20 ng/uL
                respectively; those concentrations are equivalent
                to 50-uL extracts of 10-g samples containing
                1, 5, 25, and 100 ppb, respectively, or of 1 L
                water samples containing 0.01, 0.05, 0.25, and
                1.0 ppb, respectively.

       7.6.1.3  Solutions #1-04 contain 13C12-1,2,3,4-TCDD at a
                concentration of 0.6 ng/uL.

       7.6.1.4  Solutions #l-#2 contain 37ClA-2,3,7,8-TCDD at a
                concentration of 0.028 ng/uL, this concentration
                Is equivalent to an extract of a sample containing
                0.14 ug/kg or 1.4 ng/L the amount of  'Cl^-TCDD (the
                surrogate compound) added to each sample before
                extraction.

7.6,2  Alternative Two

       Four isooctane solutions containing unlabeled 2,3,7,8-TCDD  at
       varying concentrations and   C12-2,3,7,8-TCDD (the internal
       standard (CASRN 80494-19-5) at a constant concentration.
       Three of these solutions also contain 37C1^2,3,7,8-TCDD (the
       surrogate compound, CASRN 85508-50-5) at varying concentra-
       tions.  Concentration calibration solutions are to be obtained
       from the Quality Assurance Division, US EPA Environmental
       Monitoring Systems Laboratory (EMSL-LV), Las Vegas, Nevada.
       However, if not available from EMSL-LV, standards may be
       obtained from commercial sources, and solutions may be pre-
       pared in the contractor laboratory.  Traceabillty of standards
       must be verified against EPA-supplied standard solutions,
       by laboratory SOP's as required in IFB Pre-Award Bid
       Confirmations, part 2.f.(4).                          '.'.'.

       7.6.2.1  A solution of 13C12~1,2 ,3 ,4-TCDD at a concentra-
                tion of 10 ng/uL In Isooctane  is provided to be.
                added to each of the concentration calibration
                           D - 12

-------
                     (CC) solutions.  The amount  to be  added  to  each  CC
                     solution  Is given below.  This will  result  In  all
                     compounds being In the same  ratios as  Alternative
                     One but the concentrations are 9 percent lower.

                      10 ng/uL 13C12-1,2,3,4,-TCDD Solution

         CC1  - 1  mL  CC1 + 60 uL 13C12~1,2,3,4-TCDD + 40 uL  Isooctane
         CC2  - 1  mL  CC2 + 60 uL 13C12~1,2,3,4-TCDD + 40 uL  Isooctane
         CC3  - 1  raL  CC3 + 60 uL 13C12~1,2,3,4-TCDD + 40 uL  Isooctane
         CC4  - 1  mL  CC4 + 60 uL I3C12~1,2,3,4-TCDD * 40 uL  Isooctane

           7.6.2.2   The final CC//1-CC#4  solutions contain  13C12~
                     2,3,7,8-TCDD at 0.909 ng/uL  which  Is equivalent
                     to a 50-uL extract of a  10 g or 1  L  sample  to
                     which  that compound  (the Internal  standard)
            _^        was added at a concentration of 4.5  ug/Kg or
                     45 ng/L.

           7.6.2.3   The final CC#1-CC#4  solutions contain  unlabeled
                     2,3,7,8-TCDD at concentrations of  0.182, 0.909,
                     4.545, and 18.18 ng/uL,  respectively.

           7.6.2.4   Solutions #l-#4 contain  13C12-1,2,3,4-TCDD  at
                     a concentration of 0.54  ng/uL.

           7.6.2.5   Solutions //1-//3 contain  37Cl4-2,3,7,8-TCDD
                     at concentrations of 0.054,  0.109  and  0.182
                     ng/uL, respectively.

                     NOTE:  The  surrogate concentrations do  not
                           correspond to Alternative One and are not
                           at the  level  used for the confirmation
                           of the  1.0 ug/kg  or  10 ng/L detection
                           criteria.

     7.6.3  Store concentration calibration  solutions  In 1-mL amber ralni-
           vlals at room temperature.

7.7  Performance Check Solution   A mixture  containing at  a minimum:
     unlabeled 2,3,7,8-TCDD (CASRN  1746-01-6); 1,2,3,4-TCDD (CASRN
     30746-58-8); 1,4,7,8-TCDD (CASRN  40581-94-0);  1,2,3,7-TCDD (CASRN
     67028-18-6); 1,2,3,8-TCDD (CASRN  53555-02-5);  1,2,7,8-TCDD (CASRN
     34816-53-0) and 1,2,6,7-TCDD  (CASRN  40581-90-6) must be obtained from
     the Quality Assurance  Division,  Environmental Monitoring Systeris
     Laboratory, Las Vegas, Nevada.   Note:   This solution may vary between
     lots.                                                        :   '
     To this dry mixture add 500 uL of the sample fortification solution
     (Section 78) containing  3C12-2,3,7,8-TCDD at a concentration cf 0.5
     ng/uL and 37C1,-2,3,7,8-TCDD at a concentration of 0.014 ng/uL aud 50
                                D - 13                          8/87 Rev.

-------
          uL of the 10 ng/uL 13C12~12>3,4-TCDD recovery standard solution.
          Store In 1-mL amber mlnl-vlal at 4癈.

     7.8  Sample Fortification Solution  An isooctane solution containing the
          internal standard at a concentration of 0.5 ng/uL and the surrogate
          compound at a concentration of 0.014 ng/uL.  Mix 100 uL with 1.5 mL of
          acetone before adding to each sample and blank.

     7.9  Field Blank Fortification Solution  An isooctane solution con-
          taining the internal standard at a concentration of 0.5 ng/uL, the
          surrogate compound at a concentration of 0.014 ng/uL, and the unlabeled
          2,3,7,8-TCDD at a concentration of 0.1 ng/uL.  Mix 100 uL with 1.5 raL
          of acetone before adding to each field blank.

    7.10  Recovery Standard Solution  An isooctane solution containing the
          recovery standard   Ct0-1,2,3,4-TCDD at a concentration of 10 ng/uL.

          7.10.1 ""For samples to be analyzed using Alternative One CC Solu-
                  tions, the recovery standard   C^2~1,2,3,4-TCDD is used at a
                  concentration of 10 ng/uL.

          7.10.2  For samples to be analyzed using Alternative Two CC solutions
                  as standards, the recovery standard 13C12-1,2,3,4-TCDD mus!:
                  be diluted before use; this is done by adding 100 uL of
                  isooctane to a measured 1.0 mL of the 10 ng/uL   C^2~l>2,3,4-
                  TCDD solution.

8.  SAMPLE PRESERVATION AND HANDLING

    8.1  Chain-of-custody Procedures (see Exhibit G)

     8.2  Sample Preservation

          8.2.1  Soil Samples:  When received, each sample will be contained in
                 a 1-pint glass jar surrounded by vermiculite in a sealed metal
                 paint can.  Until a portion is to be removed for analysis,
                 store the sealed paint cans in a locked limited-access area
                 where ambient temperature is maintained above freezing.  After
                 a portion is removed for analysis, return the unused portion
                 of sample to its original containers and store as stated
                 above.  Do not freeze samples; they may contain sufficient
                 water to break the sample jar  if frozen.

          8.2.2  Water Samples:  Each water sample received will consist of
                 two  (2) 1 liter (or quart) amber glass  bottles.  Samples.
                 may  be iced or refrigerated at 4癈  from the  time of
                 collection until extraction.   Do not  freeze.  Samples  'ajsL  be
                 extracted within 10 days of VTSR.

         8.2.3   All  samples must be protected  from  light from the  time of
                 collection until extraction to prevent  photodecompositicn.


                                     D -  14

-------
8.3  Sample Handling

     8.3.1  CAUTION:  Finely divided soils contaminated with 2,3,7,8-TCDD
                      are hazardous because of the potential for inha-
                      lation or ingestion of particles containing 2,3,7,8-
                      TCDD.  Such samples should be handled in a confined
                      environment (i.e., a closed hood or a glove box).

     8.3.2  Pre-extraction Sample Treatment

            8.3.2.1 For the purpose of this contract a water sample is
                    defined as a single phase system, the primary
                    component of which is water.  This may include
                    floating, suspended, and settled particulate matter
                    in quantities that do not cause severe problems with
                    the extraction.  If sufficient particulate matter is
                    present to be considered a separate phase or it
                    causes severe extraction problems precede to Section
                    8.3.2.4.

            8.3.2.2 For the purpose of this contract a soil/sediment
                    sample is defined as a single phase solid system
                    composed of soil or sediment.  It may contain
                    particulates such as stones, vegetation, etc. but
                    should not contain an obvious liquid phase.  If a
                    liquid phase is present, precede to Section 8.3.2.A.
            8.3.2.3 Homogenization  Although sampling personnel will
                    attempt to collect homogeneous samples, the contractor
                    shall examine each sample and judge if it needs fur-
                    ther mixing.  NOTE:  Contractor personnel have the
                    responsibility to take a representative sample
                    aliquot this responsibility entails efforts to make
                    the sample as homogeneous as possible.  Stirring is
                    recommended when possible.

            8.3.2.4 Centrifugation  If a soil or water sample contains
                    more than one phase, contact your DPO to determine
                    which phase(s) should be analyzed.  If the sample
                    contains obvious aqueous/solid phases, centrifuge it
                    to separate liquid and solid phases (an organic
                    phase is beyond the scope of this method, contact
                    your DPO for instructions).  Place the entire Cample
                    in suitable centrifuge bottle(s) and centrifuge Ifor
                    30 minutes at 400 x G.  Remove bottle(s) from
                    centrifuge and decant the aqueous phase to be :  .  .
                    analyzed as a water sample.  Mix solid layer
                    with stainless steel spatula and remove a portion co
                    be weighed and analyzed as a soil/sediment sample.
                                D -  15

-------
                          Return the remaining solid portion to original sample
                          bottle and store.

                          CAUTION:   A phase  not analyzed may contain TCDD and
                                    should be handled and disposed of
                                    appropriately.
9.   CALIBRATION
     9.1  Two types of calibration procedures are required.   One type, initial
          calibration, is  required before any samples are analyzed for TCDD,
          and is required  intermittently throughout sample analyses as dictated
          by results of routine calibration procedures described below.  The
          other type, routine calibration, consists of analyzing the column
          performance check solution and concentration calibration solution //I
          (Section 7.6).  No samples are to be analyzed until acceptable cali-
          bration as described in Section 9.2 and 9.3 is demonstrated and
          docuraenFed.

     9.2  Initial Calibration

          9.2.1  Concentration calibration solutions  the  four solutions
                 described in Section 7.6 are required.

          9.2.2  Inject an appropriate aliquot of the performance check
                 solution  (CAUTION:  See Section 6.1.1) and  acquire selected-
                 ionmonitoring (SIM) mass spectral data using the MS operating
                 conditions specified in Section 9.2.4.  Determine GC operating
                 conditions necessary to achieve separation  described in
                 Section 9.2.6.1.

          9.2.3  Determine valley percent as described in Section 9.2.6.1 and
                 the m/z ratio according to the criteria in  Section 9.2.6.2.
                 If the valley percent and/or m/z ratios are outside require-
                 ments, corrective action to meet the criteria must be taken
                 (as described in Section 6.1.1) before further sample analyses
                 are performed.

          9.2.4  Using the same GC conditions that produced  acceptable results
                 with the  performance check solution, analyze a 2-uL or other
                 appropriate aliquot (as described in Section 6.1.1) of each of
                 the four  concentration calibration solutions with the
                 following MS operating parameters.

                 9.2.4-1  Acquire selected-ion-monitoring data for m/z 259.
                          320, 322, 328, 332 and 334.                     '.

                 9.2.4.2  Total cycle time for data acquisition must be'<1.3
                          seconds.

                 9.2.4.3  Acquire at least five data points for each ion during
                          elution of the GC peak.

                                     D - 16

-------
       9.2.4.4  Use the same data acquisition time foe each of the
                six ions being monitored.

9.2.5  Repeat Section 9.2.4 two times to produce triplicate data sets
       for each solution.  NOTE: CC solutions should be analyzed In
       either random order or in order of increasing concentration to
       avoid biasing the calibration.

9.2.6  The Laboratory must not proceed with analysis before deter-
       mining and documenting acceptable calibration with the
       following criteria:

       9.2.6.1 GC Column Performance

               9.2.6.1.1  The valley between 2,3,7,8-TCDD and the
                          peaks representing all other TCDD Isomers
                          must be resolved with a valley 25%.
       ""                Valley (%) = x/y X 100, when y is peak
                          height of 2,3,7,8TCDD, x Is measured as
                          shown in Figures 2 and 3 at the end of this
                          Exhibit.   The peak representing 2,3,7,8-
                          TCDD shall be labeled and identified as
                          such on the chromatograms.

               9.2.6.1.2  Ratio of integrated ion current for m/z 320
                          to m/z 322 for 2,3,7,8-TCDD must be >0.67
                          and 0.90.

               9.2.6.1.3  Ratio of integrated ion current for m/z 332
                          to ra/z 334 for 13C12-2,3,7,8-TCDD must be
                          X).67 and 0.90.

       9.2.6.2  Calibration solutions must meet the following
                criteria:

                9.2.6.2.1 MS sensitivity  signal-to-noise (S/N)
                          ratio (Section 3.10) of >2.5 for m/z
                          259, 320,and 322 for unlabeled 2,3,7,8-
                          TCDD and 328 for 37Cl4-2,3,7.8-TCDD and
                          >10 for ra/z 332 and 334 for I3C12-2,3,7,8-
                          TCDD.

                9.2.6.2.2 The ratio of integrated ion current for m/z
                          320 to m/z 322 for 2,3,7,8-TCDD must be
                          >0.67 and 0.90.

                9.2.6.2.3 The ratio of integrated ion current 'fot. m/z
                          332 to m/z 334 for 13C12-2,3,7,8-TCDD must
                          be >0.67 and <0.90.
                           D -  17

-------
 9.2.6.3  Calculate the response factor for unlabeled 2,3,7,8-TCDD
          relative to 13C122,3,7,8-TCDD:
                                Ax-
                      RRFn  =
                               Als -
        Where  Ax  =  the sura of integrated ion abundances of m/z
                      320 and m/z 322 for unlabeled 2,3,7,8-TCDD,

              A^s  =  the sura of integrated abundances of m/z
                      332 and m/z 334 for 13C12-2,3,7,8-TCDD,

              Qls  =  quantity of 13C12-2,3,7,8-TCDD, and

               Qx  =  quantity of unlabeled 2,3,7,8-TCDD
                      injected.

              RRF is a diraensionless number; units used to
              express quantities must be consistent.

9.2.6.A  Calculate the response factor for 13C12-2,3,7,8-TCDD
         relative to 13C12-1,2,3,4-TCDD:

                 Ais - Qrs
              =  	
                 A
         lrs *

Where A
               rs  =  The sura of the integrated ion abundance of
                      m/z 332 and m/z 334 for 13C12-1,2,3,4-TCDD

              Qrs  =  Quantity of 13C12-1,2,3,4-TCDD Injected
                      A and Q are as in Section 9.2.6.3.

 9.2.6.5  Response Factor Criteria:
          9.2.6.5.1  Calculate the mean RRF and its percent relative
                     standard deviation (%RSD) from triplicate
                     analysis of each of 4 concentration solutions
                                                       12-
             for unlabeled 2,3,7,8-TCDD and 13C17-TCDD.
                           Standard Deviation
                  %RSD  =  	  x 100
                               Mean RRF
                      D  -  18

-------
                     9.2.6.5.2  The variation of the RRF calculated for
                                unlabeled 2,3,7,8-TCDD at each
                                concentration level must not exceed 10%
                                RSD.

                     9.2.6.5.3  Calculate the mean and %RSD of the 4 mean
                                RRFs for unlabeled 2,3,7,8-TCDD and for
                                13C12-TCDD.

                     9.2.6.5.4  The %RSD of  the 4 mean RRFs for 13C12~
                                2,3,7,8TCDD  should not exceed 10% RSD.

                     9.2.6.5.5  The %RSD of  the 4 mean RRFs for unlabeled
                                TCDD must not exceed 10% RSD.

                     9.2.6.5.6  The mean of  the mean RRFs for each com-
                                pound must be used for concentration
                                calculations.

                     9.2.6.5.7  The concentration curves must be plotted
                                (RRF vs concentration) for enclosure in
                                the deliverables package.

9.3  Routine Calibration

     9.3.1  Inject an appropriate aliquot (CAUTION:  See Section 6.1.1)
            of the performance check solution (Section 7.7) and acquire
            selected ion monitoring mass spectral data for m/z 259, 320,
            322, 328, 332, and 334 within a total cycle time of <1.5
            seconds.  Acquire at least five data points for each GC peak
            and use the same data acquisition time for each of the six
            ions being monitored.  NOTE:  The same data acquisition para-
            meters previously used to analyze concentration calibration
            solutions during initial calibration must be used for the
            performance check solution.  The column performance check
            solution must be run at the beginning and end of each 12-hour
            period, if the contractor laboratory operates during consecu-
            tive 12-hour periods (shifts), analysis of the performance
            check solution at the beginning of each 12-hour period and at
            the end of the final 12-hour period  is sufficient.

     9.3.2  Determine and document  acceptable column performance as
            described in Section 9.2.6.1.

     9.3.3  Inject 2 uL of concentration calibration solution #1 which
            contains 0.2 ng/uL of unlabeled 2,3,7,8-TCDD once at  the
            begininning of each  12-hour period.   Using  the  same GC'rfS/DS
            conditions as used in Section 9.3.1,  acquire data  for  m/z
            259, 320, 322, 328, 332 and 334.  Determine  and document
            acceptable calibration  as described  below.
                                 D - 19

-------
                 9.3.3.1  MS sensitivity  signal-to-noise (S/N) ratio (Sec-
                          tion 3.8) of >2.5 for m/z 259, 320, 322,and 328 for
                          unlabeled 2,3,7,8-TCDD and 37Cl4-2,3,7,8-TCDD and MO
                          for m/z 332 and 334 for 13C l2-2,3,7,8-TCDD is
                          required.  The ratio of integrated ion current for
                          m/z 320/322 must be X).67 and <0.90.

                 9.3.3.2  Measured response factor for unlabeled 2,3,7,8-TCDD
                          relative to 13C12-2,3,7,8-TCDD must be within +10% of
                          the mean value established (Section 9.2.6.5.3) by
                          initial analyses of the concentration calibration
                          solutions.

         9.3.4  Further sample analyses must not be performed if the criteria
                in Section 9.3.2 and 9.3.3 are not met.  Possible remedies are
                listed in Section 9.3.5.  Following corrective action,  a
                'routine calibration must be performed and criteria listed in
                Section 9.3.3 must be met before further analysis of samples is
                performed; if the routine calibration does not meet criteria
                a new initial calibration must be performed.

         9.3.5  Remedial actions shall be taken by Contractor if criteria are
                not met.  Possible remedies are:

                9.3.5.1  Check and adjust GC and/or MS operating conditions.

                9.3.5.2  Replace GC column (performance of initial calibration
                         procedures are required if acceptance criteria for
                         continuing calibration are not met).

                9.3.5.3  Tune MS for greater or lesser resolution.

                9.3.5.4  Calibrate MS mass scale.

                9.3.5.5  Prepare and analyze new performance check solution.

10.  QUALITY CONTROL

     See Exhibit E for QA/QC Requirements.

11.  PROCEDURES

     11.1 Soil Sample Extraction

         11.1.1  CAUTION:  See Section 5 for safety guidelines and recommendations,

         11.1.2  Jar extraction.  NOTE: Extremely wet samples may require
                 centrifuging to remove water before addition of sodium
                 sulfate (see Section 8.3.2.2).
                                     D - 20                         Rev, 8/87

-------
11.1.2.1  Accurately weigh to three significant figures a
          10 g (+0.5 g)  portion of the wet soil or sediment
          sample,  and transfer It to the extraction jar.

11.1.2.2  Note:   Additional QC samples are required as speci-
          fied In exhibit E.   These are processed as described
          In this section with the following exceptions:

               a.   Laboratory Method Blank - Perform all steps
                   In the analytical procedure but substitute
                   an aliquot of sodium sulfate for the soil/
                   sediment sample.

               b.   Fortified Field Blank - Perform all steps
                   In the analytical procedure, but spike the
                   designated sample with 1.5 raL of the acetone
                   dilution of the field blank fortification
                   solution (Section 7.9) rather than the sample
                   fortification solution.

11.1.2.3  Add 1.5 mL of  the acetone dilution of the sample
          fortification  solution (Section 7.8) to the soil or
          sediment in the extraction jar.  Add small portions
          of the solution at several sites on the surface of
          the soil or sediment.

11.1.2.4  Add 20 g of purified anhydrous sodium sulfate, and
          mix thoroughly using a stainless steel spoon or
          spatula.

11.1.2.5  Allow the mixture of soil and sodium sulfate to set
          for 2 hours at ambient temperature; mix again,
          break all visible lumps, and allow to set for at
          least A more hours.

11.1.2.6  Mix again and  add 20 mL of methanol; mix again and
          add 150 mL of  hexane.

11.1.2.7  Place the extraction jar containing the soil,
          sodium sulfate and solvents in the shaker and shake
          for at least 3 hours.

11.1.2.8  Remove the jar from the shaker and allow all solids to
          settle.  Decant the solvent through a glass funnel
          containing hexane-rinsed filter paper into a clean
          Kuderna-Danish apparatus.  Rinse the jar, solid
          sample  residue, and  filter residue with four ;J-mL
          portions of hexane.
                     D - 21

-------
           11.1.2.9  Concentrate the extract  volume to approximately 2
                     to 3 mL with a Kuderna-Danlsh apparatus.   NOTE:
                     Glassware used for more  than one sample must be
                     carefully cleaned between samples to prevent cross
                     contamination (see Section 6.5).

           11.1.2.10  Rinse the evaporator flask with 3 mL portions of
                      hexane;  transfer each rinse to the  concentrator
                      tube.  Between additions of hexane rinse,  reduce the
                      extract volume in the concentrator tube enough to
                      allow addition of another 5 mL volume of rinse.   To
                      reduce the volume,  place the concentrator tube in a
                      water bath adjusted to  operate at 50癈 and position
                      the tube so that the surfaces of the extract and the
                      water are at about  the  same level.   Evaporate the
                      solvent with a stream of nitrogen with the tip of the
                      nitrogen delivery tube  2 cm above the solution.

           ini.2.11  After the final rinse has been added, reduce the
                      extract volume to approximately 1 mL.  Proceed to
                      section 11.3.2.  If further processing will be
                      delayed, quantitatively transfer the extract to a
                      Teflon sealed screw-cap vial and store refrigerated
                      and protected from light.

11.2  Water Sample Extraction

      Caution:   When using this method to analyze for 2 ,3 ,7 ,8-TCDD, all
                of the following operations should be performed in a
                limited access laboratory with the analyst wearing full
                protective covering for all exposed skin surfaces.  See
                Section 5.

    11.2.1  Mark the water meniscus on the side of the sample bottle for
            later determination of the sample volume.  Pour the entire
            sample into a 2 L separatory funnel.   Note: a continuous
            liquid-liquid extractor may also  be used.

            11.2.1.1.  NOTE:  Additional QC samples are required
                       as specified in exhibit E.  These are
                       processed as described in this section with
                       the following exceptions:

                           a.  Laboratory Method Blank - Perform all steps
                               in the analytical procedure but substitute
                               an aliquot of reagent water for the .??nple.
                           b.  Fortified Field Blank - Perform all 's
                               in the analytical procedure, but spike
                               the designated sample with 1.5 mL of . Lh-i
                               acetone dilution of the field blank  forti-
                               fication solution (Section 7.9) rather than
                               the sample fortification solution.

                                D - 22

-------
 11.2.2  Add 1.5 mL of the acetone dilution of the sample fortification
         solution containing 50 ng of the Internal standard and 1.4 ng
         of the surrogate compound to the sample in the separatory
         funnel.

 11.2.3  Add 60 mL of raethylene chloride to the sample bottle,  seal,
         and shake 30 seconds to rinse the inner surface.   Transfer the
         solvent to the separatory funnel and extract the sample by
         shaking the funnel for 2 minutes with periodic venting to
         release excess pressure.  Allow the organic layer to separate
         from the water phase for a minimum of 10 minutes.  If  the
         emulsion interface between layers is more than one梩hird the
         volume of the solvent layer, the analyst must employ mechan-
         ical techniques to complete the phase separation.  The optimum
         technique depends upon the sample, but may include stirring,
         filtration of the emulsion through glass wool, centrifugation,
         or other physical methods.  Collect the methylene chloride
         extract in a 250 mL Erlenmeyer flask.

11.2.4   Add a second 60 mL volume of raethylene chloride to the sample
         bottle and repeat the extraction procedure a second time,
         combining the extracts in the Erlenmeyer flask.  Perform a
         third extraction in the same manner.

11.2.5   Assemble a Kuderna-Danish (K-D) concentrator by attaching a 10
         mL concentrator tube to a 500 mL evaporative flask.  Pour the
         combined extracts into the K-D concentrator through a drying
         column containing about 6 cm of sodium sulfate.  Rinse the
         Erlenmeyer flask with 25-30 mL methylene chloride and pour it
         through the drying column, rinse the drying column with an
         additional 10 ml methylene chloride.  All rinses are added to
         the concentrator.
                                         %
11.2.6   Add one or two clean boiling chips to the evaporative flask
         and attach a three-ball Snyder column.  Prewet the Snyder
         column by adding about 1 mL of methylene chloride to the top.
         Place the K-D apparatus on a hot water bath (60 to 65癈) so
         that the concentrator tube is-partially immersed in the hot
         water, and the entire lower rounded surface of the flask is
         bathed with hot vapor.  Adjust the vertical position of the
         apparatus and the water temperature as required to complete
         the concentration in 15-20 minutes.  At the proper rate of
         distillation, the balls in the column will actively chatter
         but the chambers will not flood with condensed solvent.  .When
         the apparent volume of liquid reaches 1 mL, remove the -K.-D
         apparatus and allow it to drain and cool for at  least 10
         minutes.

11.2.7   Momentarily remove the Snyder column, add 50 mL  of hexare  and
         a new boiling chip, and reattach  the Snyder column.  Raise  the
                             D - 23

-------
              temperature  of  the water  bath  to  85  to  90癈.   Concentrate  the
              extract  as  In section 11.2.6 except  use hexane to  prewet the
              column.   Remove the Snyder  column and rinse  the flask and  Its
              lower joint  Into the concentrator tube  with  1  to 2 mL of
              hexane.

     11.2.8   Add a clean  boiling chip  to the concentrator tube  and attach a
              two-ball mlcro-Snyder column.  Prewet the  column by adding
              about 1  raL of hexane to the top.   Place the  mlcro-K-D
              apparatus on the water bath so that  the concentrator tube  Is
              partially immersed in the hot  water.  Adjust the vertical
              position of  the apparatus and  the water temperature as
              required to  complete the  concentration  In  5  to 10  minutes.
              At the proper rate of distillation the  balls of the column
              will actively chatter but the  chambers  will  not flood. When
              the apparent volume of the  liquid reaches  about 0.5 mL,
              remove the K-D  apparatus  and allow it to drain and cool for
              at least 10  minutes.  Remove the  raicro-Snyder column and
              rinse its lower joint into  the concentrator  tube with 0.2  mL
              hexane.   Proceed to section 11.3.2.   If further processing
              is to be delayed, the extract  should be quantitatively
              transfered  to a Teflon sealed  screw-cap vial and store
              refrigerated and protected  from light.

     11.2.9  Fill the sample  bottle with  water  to  the mark and measure  the
             volume to the nearest 10 mL  In  a 1 L  graduated cylinder.

11.3  Column Chromatograph

     11.3.1 Column Preparation

            11.3.1.1  Column  1:  Place  1.0 g of silica gel into  a 1 cm  x
                      20 cm column and  tap  the  column gently to  settle
                      the silica gel.   Add  2 g  sodium hydroxide-impregnated
                      silica gel, 1 g  silica gel,  4.0 g of sulfuric acid-
                      impregnated silica  gel,  and 2 g silica gel.  Tap
                      column gently after each addition.

            11.3.1.2  Column 2:  Place  6.0  g of alumina into a 1 cm x 30
                      cm column and tap the column gently to settle the
                      alumina.  Add a 1-cm layer of purified sodium
                      sulfate  to the top  of  the alumina.

            11.3.1.3  Add hexane to each column until the packing  is free
                      of channels and air bubbles.  A small positive 
                      pressure  (5 psi)  of clean nitrogen can be  used if
                      needed.

     11.3.2  Quantitatively  transfer the hexane sample  extract  from rh
             concentrator  tube  to  the  top of the silica gel  in Column 1.
             Rinse  the concentrator tube with  two 0.5 mL  portions  el
             hexane;  transfer  rinses to Column 1.

                                  D - 24

-------
     11.3.3  With 90 mL of hexane,  elute the extract from Column 1 directly
             Into Column 2 containing alumina and sodium sulfate.

     11.3.4  Add 20 mL of hexane to Column 2 and elute until the hexane
             level Is just below the top of the sodium sulfate;  discard the
             eluted hexane.

     11.3.5  Add 20 raL of 20% methylene chloride/80% hexane (volume/volume)
             to Column 2 and collect the eluate.

     11.3.6  Reduce the volume of the eluate with a gentle stream of
             filtered dry nitrogen.   When the volume of the eluate is about
             1 to 2 mL, transfer the eluate to the Carbopack column
             (Section 11.4.4).  Rinse the eluate container with  two 0.5
             raL portions of hexane;  transfer the rinses to the Carbopack
             column.  CAUTION:  Do  not evaporate the sample extract to
             dryness.  NOTE:  The carbopack cleanup is not required for
             water samples unless needed to meet detection sensitivity
             criteria.

11.4  Carbopack Column Chromatography Procedure

     11.4.1  Thoroughly mix 3.6 g of Carbopack C (or equivalent) with
             16.4 g of Celite 545 (or equivalent) in a 40 raL vial and
             activate by heating in an oven at 130癈 for 6 hours.
             Store in a desiccator.   CAUTION:  Check each new batch of
             mixed Carbopack/Celite to ensure TCDD recovery of >50%.
             Subject the low level  concentration calibration solution to
             this procedure and measure the quantity of labeled  and
             unlabeled 2,3,7,8-TCDD.

     11.4.2  Insert a small plug of glass wool into a disposable
             pipet approximately 15 cm long by 7 mm O.D.  Apply
             suction with a vacuum aspirator attached to the pointed
             end of the pipet, and add the Carbopack/Celite mixture
             until a 2 cm packing is obtained.

     11.4.3  Pre-elute the column with:

             11.4.3.1  2 mL toluene

             11.4.3.2  1 mL of mixture of 75% (by volume) methylene
                        chloride, 20% methanol and 5% benzene

             11.4.3.3  1 mL of 50%  (by volume) cyclohexane and 50%
                        methylene chloride

             11.4.3.4  2 mL of hexane

     11.4.4  While  the column  is still wet with hexane add the sample
             extract  from section 11.2.6.  Elute the column with  the
             following sequence of  solvents and discard  the eLaates.

                                 D  - 25

-------
             11.4.4.1  2 mL hexane

             11.4.4.2  1 mL of 50% (by volume)  cyclohexane and  50% methylene
                       chloride

             11.4.4.3  1 mL of 75% (by volume)  methylene chloride, 20%
                       methanol and 5% benzene

     11.4.5  Elute with 2 mL of toluene and collect the elutate,  which
             contains the TCDD.  Transfer the rinses to a 1-mL  amber  mini-
             vial with conical reservoir with further concentration as
             necessary.  CAUTION:  Do not evaporate the sample  extract to
             dryness.

     11.3.6  Store the sample extract in the dark at 4癈 until  just before
             GC/MS analysis.

11.5  GC/MS Analysis

     11.5.1  Remove the sample extract or blank from storage and  allow it
             to warm to ambient laboratory temperature.  With a stream of
             dry, filtered nitrogen, reduce the extract/blank volume  to
             near dryness.  Immediately before  GC/MS analysis,  add 5  uL of
             the 10 ng/uL recovery standard solution and adjust the extract
             or blank volume to 50 uL with isooctane.

     11.5.2  Inject a 2-uL aliquot of the extract into the GC,  operated
             under conditions previously used (Section 9) to,produce
             acceptable results with the performance check solution.

     11.5.3  Acquire mass spectral data for the following selected
             characteristic ions:  m/z 259, 320, and 322 for unlabeled
             2,3,7,8-TCDD: m/z 328 for 37Cl4-2,3.7,8-TCDD; and  m/z 332
             and 334 for 13C,-,-2,3,7 ,8-TCDD and 13C, 7-l ,2 ,3 ,4-
                           >i 2
             TCDD.  Use the same data acquisition time and MS operating
             conditions previously used (Section 9.2.6) to determine
             response factors.

11.6  Identification Criteria.  NOTE: Refer to Exhibit E,  Section 7, for
      application of identification criteria.

     11.6.1  Retention time (at maximum peak height) of the sample com-
             ponent must be within 3 seconds of the retention time of
             the 13C12-2,3,7,8-TCDD.  Retention times are  required for
             all chromatograms, but scan numbers are optional.  These
             parameters should be printed next to the appropriate peak.

     11.6.2  The integrated ion currents detected for m/z  259, 320, ana
             322 must maximize simultaneously.  If there are peaks thtt
             will affect the maximization or quantitation  of peaks of
             interest, attempts should be made to narrow the scan window
             to eliminate the interfering peaks.  This should be report.^d
             on a separate chromatograra.

                                 D - 26

-------
         11.6.3  The integrated ion current for each analyte ion (m/z 259,
                 320 and 322) must be at least 2.5 times background noise and
                 must not have saturated the detector; internal standard ions
                 (m/z 332 and 334) must be at least 10 times background noise
                 and must not have saturated the detector.

         11.6.4  Abundance of integrated ion counts detected for m/z 320 must
                 be MJ7% and 7% and <90% of integrated ion counts detected for ra/z
                 334.

         11.6.6  The recovery of the internal standard 13C12-2,3,7,8-TCDD should
                 be within a 40 percent to 120 percent recovery window.  This
                 is an advisory limit only, an action window may be set when
                 sufficient data is available.

12. CALCULATIONS

    12.1 Concentration

         12.1.1  Calculate the concentration of 2,3,7,8-TCDD using the formula:

                                    AX  Qis

                           癤 ~   Ais . RFn . W

                where      Cx  =  2,3,7,8-TCDD concentration in ug/kg or ug/L

                           AX  =  the sum of integrated ion abundance
                                  detected for m/z 320 and 322

                          AIS  =  the sum of integrated ion abundances
                                  detected for m/z 332 and 334
                                  (characteristic ions of
                                  I3C12-2,3,7,8-TCDD,  the internal
                                  standard)

                          Qis  =  quantity (in ng) of  13C^-2,3,7,8-TCDD added
                         RRF
      to the sample before extraction

n  =  calculated mean response factor for
      unlabeled 2,3,7,8-TCDD relative to
      l3C12-2,3,7,8-TCDD

W  =  weight (in g) of wet soil or sediment sample
      or volume of water extracted (in mL).
                                     D - 27                            Rev. 8/87

-------
     12.1.2   If the calculated concentration  of  unlabeled  2,3,7,8-TCDD
             exceeds 100 ug/kg for soil/sediment or 1  ug/L for water,  which
             Is the maximum concentration of  the concentration calibration
             solutions,  the linear range may  have been exceeded,  and a
             smaller aliquot of that  sample must be analyzed.   Accurately
             weigh to three significant figures  a 1-g  aliquot  of  the wet
             soil/sediment  or measure a 100 mL aliquot of  water.   Add the
             1.5 mL acetone dilution  of 100 uL of the  sample fortification
             solution (Section 7.8),  just as  for the larger sample aliquot.
             Extract and analyze.

             12.1.3  Calculate the concentration of the internal  standard
                     l3C12-2,3,7,8-TCDD using the formula:
                               Ais   Q
                                      rs
                     Cis
                             Ars  . RFi  . W
             where

                     C,    =  concentration  of  13C, 9-2,3,7,8-TCDD in ug/kg
                                  .               i
                             or ug/L

                     Ms  =  sum of  integrated ion abundances  for m/z
                             332 and 334  for 13C12-2,3,7,8-TCDD

                     Ars  =  sum of  integrated ion abundances  for m/z
                             332 and 334  for 13C12-1,2 ,3,4-TCDD

                     Qrs  =  quantity (in ng)  of  13C12~1 ,2,3,4-TCDD added
                             to the  sample  before injection

                     RF                                          1 3
                       i  =  calculated mean response factor for   C]o~
                             1,2,3,4-TCDD

                       W  =  weight  (in g)  of  wet soil or sediment
                             sample  or volume  of  water extracted (in
                             mL).

12.2 Estimated Maximum Possible Concentration    For samples in which no
     unlabeled 2,3,7,8-TCDD was detected, calculate the estimated
     maximum possible concentration, which is  the concentration required
     to produce a signal with peak height of  2.5  times the background
     signal level.  The background level  is determined by measuring tne
     range of the noise (minimum to maximum)  for  either m/z 320 or. 322
     in the appropriate region of the SICP (as defined in section I'
     3.11), multiplying that noise height by  2.5, and relating the
     product height to an estimated concentration that would produce
     that product height.

     Use the formula:
                                 D - 28

-------
                          2.5    Hx
                  MFC =
                           His  .  RFn . W
     where  MFC  =  estimated maximum possible concentration of unlabeled
                    2,3,7,8-TCDD required to produce tfx in ug/kg or ug/L

             Hx  =  peak height for either m/z 320 or 322 within +_ 5 scans
                    of the internal standard peak used to measure H^s

            His  =  peak height of the appropriate ion characteristic of
                    the internal standard m/z 332 when m/z 320 is used
                    to determine Hx,  and m/z 334 when m/z 322 is used
                    to determine Hx

            -Qis, RF and W retain the definitions previously stated in
            Section 12.1.1

12.4  The relative percent difference (RPD) is calculated as follows:
      (See Section 5.1.1, Exhibit E.)

      RPD  = |  sl - S2 |  x 100 .  =        |sl   S2 |  x 100

             Mean Concentration             sl + S2

                                               2

      **1 and ^2 represent sample and duplicate sample results.

12.6  Percent Recovery of 2,3,7,8-TCDD in spiked field blanks =

      concentration found
      	x 10o
      concentration added

12.7 . Percent Recovery of internal standard, 13C12-2,3,7,8-TCDD  =

   ... -  concentration found
      	  x
      concentration added
                                   /N (Xt - X)2
12.8  Standard deviation  =  S  = A/ 2 ---------
                                  Vi=l  N - 1
12.9  Percent relative standard deviation  =

      Standard Deviation              S
      ------------------   x 100  =   -  x 100
            Mean                      X
                                 D - 29                           Rev. 8/87

-------
                    TABLE 1.  OPERATING CONDITION GUIDELINES
Column coating

Film thickness

Column dimensions

Helium* linear velocity


Initial temperature

Initial time

Temperature program
2,3,7,8-TCDD retention
 time
SP-2330

0,2  urn

60 m x 0.24 mm

28-29 cm/sec
at 240癈

70癈

4 min

Rapid increase to 200癈
200癈 to 240癈
at 4癈/min

24 min
CP-SIL 88

0.22 urn

50 m x 0.22 mm

28-29 cm/sec
at 240癈

45癈

3 min

Rapid increase to 190癈
190癈 to 240癈
at 5癈/min

26 min
*Hydrogen is an acceptable carrier gas.
                                     D -  30

-------
TABLE 2.  COMPOSITION OF CONCENTRATION CALIBRATION SOLUTIONS


Solution //
CC1
CC2
CC3
CCA

These are the
2,3,7,8-TCDD,

Solution #
CC1
CC2
CCS
CCA
Alternative One
(TCDD)
Unlabeled
2,3,7,8 J7Cl4-2,3,7,8 1JC12-2,3 ,7 ,8 13C12~1 ,2 ,3 ,A
0.2 ng/uL 0.028 ng/uL 1.0 ng/uL 0.6 ng/uL
1.0 0.028 1.0 0.6
5.0 	 1.0 0.6
20.0 	 1.0 0.6
Alternative Two
37
final concentrations obtained. All compounds except Cl.-
are in the same ratios as Alternative One but are 9% lower.
(TCDD)
Unlabeled
2,3,7,8 37C142,3,7,8 13C12-2 ,3 ,7 ,8 13C12~1 ,2 ,3 ,A
0.182 ng/uL 0.05A ng/uL 0.909 ng/uL 0.5A5 ng/uL
0.909 0.109 0.909 0.5A5
A.5A5 0.182 "0.909 0.5A5
18.18 	 0.909 0.5A5
                           D - 31

-------
                         MEASUREMENT OF SIGNAL TO HOISE RATIO
l/l

Cl
a:
n
z
D
I
v^

I-

b!

a:

u

z
o
J.5 -
2.5 -
  2 -
 .5 -
0.5  -
  0 -
                                      SCAM NUMBER
                    Figure 1.  Measurement of signed  to noise ratio.

-------
LJ
U)
                   lee.e
MID R1C                         DATA) 860X606 II
86/85/86  7i 12:89                 CALli 85FC436I1A II
SAI1PLEI TCDO CO.UI1H PERFORMANCE CHECK HIX
COIIOS.I CC PROGRAM SAME AS DIOXIII 1FB  (SP-2338)
RAIICEt C  1,2418  LABELi II  8, 4.8 QUAIIi  fl  8, 1.8 J  8  BASEi U 28,  3
                1842

                  - 2,3.7.8-TCDO
                        1859
                                                  A- 1,2,3,4-TCDO
                                                      1,2,3,7-TCOO

                                                   -  1.2,3,8-TCDD
                                                                                SCANS  1799 TO 2888
                    RIC
                                                                                                                   332888.
                                                                                  19.31
                                                                                    r  1.2.7,8-TCDO
                                                             1887, 1981  19J 2
                                                                                                    1974
                                                                                                     \- 1.2,6.7-TCDO
                                                                                   1993
                      1808
                   1838
                   27i88
1988
2? 152
1358
28i3S
2888  SCrtl
29)28 1UE
      Figure 2.   Selected:  ion current profile for m/z 320  and  322  produced  by MS analysis of  performance  check
                   solution  using  a 60-m SP-2330 fused  silica capillary  column and conditions listed in  Table 1.

-------
o
U)
                      iee.e-i
                      RtC
                                                           OATAi 860X879 II        SCfWS 1868 TO 2169
                             ee/94/e? i!ti7iP9                cm.it 86FC43694 is         '
                             SAMPLE! TCDO COLUTBI PERFORrtftllCE CHECK MIX
                             COUDS.t McTHCO 2                            ...
                             RAHCEt C   I.  36  LABEL: H 6, 4.9  QUAIIi A  8, 1.9 J  8  BASE) U 26.  3
                                                 - 2,3.7.8-TCOO
                                1,4,7,8-tf.DO  -
                         1809
                         24(66
1859
24149
                                                          -  1,2.3.4-TCDO
                                                            1,2.3,7-TCDO

                                                            1,2.3,8-TCOD
                                                          ^	*~~.
1369
23! 28
1956
2Si66
                                                  V1.2.7.8-TCDO
2966
26:48
2856
27:29
                                                                              429558.
                                                                                                                - 1.2,6.7-TCOD
2109  SCAII
28:88 TIME
      i-'igi-re  -3.  'Sele'-.ted  ion  current profile for m/z 320 and 322  produced  by  MS analysis of  performance  check
                    solution  using a  50-m  CP-SIL 88  fused silica capillary column and  conditions listed  in Table  1

-------
REFERENCES

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

2.  "OSHA Safety and Health Standards, General Industry," (29CFR1910), Occupa-
    tional Safety and Health Administration, OSHA 2206 (Revised, January
    1976).

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

4.  Method 613, "2,3,7,8-Tetrachlorodibenzo-p-dioxin," Federal Register, 44
    (233) 69529, December 3, 1979.

5.  "Quality Assurance Plan for 2,3,7,8-TCDD Monitoring Project,"  R.  D. Kleopfer
    and C. J. Kirchraer, presented by the Division of Environmental Chemistry,
    American Chemical Society, Washington, D.C., September 1983.

6.  "Determination of 2,3,7,8-TCDD in Soil," R. D. Kleopfer, K. Yue,  and W. W.
    Bunn, presented before the Division of Environmental Chemistry, American
    Chemical Society, Washington, D.C., September 1983.

7.  "Water Solubility of 2,3,7,8-Tetrachlorodibenzo-p-dioxin," Leland Marple,
    Robert Brunck, and Lewis Throop, Environmental Science and Technology,
    Vol. 20, No. 2, 180-182, 1986
                                      D  -  35

-------
    EXHIBIT E




QA/QC REQUIREMENTS

-------
                      Exhibit E - QA/QC Requirements


SUMMARY OF QC ANALYSES

1.  Initial and periodic calibration and instrument performance checks.

2.  Laboratory method blank analyses (Section 4.1 of QUALITY CONTROL);  minimum
    of one blank per matrix shall be analyzed with each sample batch;  an
    additional blank shall be analyzed when new reagents are used and  with each
    set of samples rerun.

3.  Analysis of a batch of samples with accompanying QC analyses:

   .  3.1  Sample Batch  <24 samples, including field blank(s),  rinsate
          sample(s) and any reruns generated by prior batch analyses.

          NOTE:  See Exhibit C, Section 3,  if total samples exceed 24,
                 additional QC analyses are required.

     3.2  Additional QC Analyses Per Batch:
          Laboratory method blank for each matrix                1-2
          Duplicate sample analysis for each matrix              1-2

          TOTAL                                                  2-4

4.  "Blind" QC samples may be submitted to contractor as an ordinary soil or
    sediment or water sample included among the batch of samples.  Blind
    samples include:

     4.1  uncontaminated soil or water,

     4.2  split samples,

     4.3  unlabeled duplicates, and

     4.4  performance evaluation samples.


QUALITY CONTROL

1.  Performance Evaluation Samples  Included among samples  in some batches
    will be samples containing known amounts of unlabeled 2,3,7,8-TCDD.

2.  Performance Check Solution and Concentration Calibration Solutions

     2.1  At the beginning of each  12-hour period during which samples are to
          be analyzed,  an aliquot of  the performance check solution and au
          aliquot of concentration  calibration solution #1 shall be analyzed to
          demonstrate adequate GC and MS resolution  and sensitivity,
          factor reproducibility, and mass range calibration.


                                      E  - 1

-------
          These procedures are described in Section 9 of Exhibit D.   If any
          required criteria are not met, remedial action must be taken
          before any samples are analyzed.

     2.2  To validate sample data,  the performance check solution must be analyzed
          at the end of each 12-hour period during which samples are analyzed.

          2.2.1  If the contractor  laboratory operates only during one 12-hour
                 period (shift) each day,  the performance check solution must
                 be analyzed twice  (at the  beginning and end of the 12-hour
                 period) to validate data  acquired during the Interim period.

          2.2.2  If the contractor  laboratory operates during consecutive 12-hour
                 periods (shifts),  analysis of the performance check solution at
                 the beginning of each 12-hour period and at the end of the
                 final 12-hour period is sufficient.

     2.3  Results"of at least two analyses  of the performance check solution
          must be reported with sample data collected during a 12-hour period.

     2.4  Deviations from criteria  specified for the performance check solution
          (Section 9.2.6.1, Exhibit D) invalidate all sample data collected
          between analyses of the performance check solution, and samples shall
          be rerun (see Exhibit C).

    The performance check mixture,  concentration calibration solutions, and the
    sample and field blank fortification solutions are to be obtained from
    EMSL-LV.  .However, if not available from EMSL-LV, standards can be obtained
    from other sources, and solutions can  be prepared in the contractor labora-
    tory.  Concentrations of all solutions  containing unlabeled 2,3,7,8-TCDD and
    not obtained from EMSL-LV must  be verified by comparison to the unlabeled
    2,3,7,8-TCDD standard solution  (concentration of 7.87 ug/mL) that is avail-
    able from EMSL-LV.
4.   Blanks
     4.1  Laboratory method blank  Perform all steps in the analytical
          procedure (Section 11, Exhibit D) using all reagents, standards,
          equipment, apparatus, glassware, and solvents that would be used
          for a sample analysis, using an aliquot of reagent water for the
          water blank and an aliquot of sodium sulfate for the soil blank.

          4.1.1  Except In the case noted below in Section 4.1,3, a laboratory
                 method blank must contain the same amount of   Cl^-2,3,7,8-VOOD
                 and 13C12-2,3,7,8-TCDD that is added to samples before
                 extraction.

          4.1.2  Extract and analyze a laboratory method blank before any
                 are extracted and analyzed.
                                     E - 2

-------
     4.1.3  Extract and analyze two laboratory method blanks before new
            solvents or reagents are used for  sample  extraction or for
            column chromatographic procedures.  Do not add any -*'C1.-
            2,3,7,8-TCDD or 13C12~2,3,7,8-TCDD to one blank, to demon-
            strate that reagents contain no impurities producing an ion
            current above the level of background noise for m/z 328, 332
            and 334.

     4.1.4  In addition to the specification in preceding section 4.1.2,
            extract and analyze a laboratory method blank for each matrix
            along with each batch of samples.

     4.1.5  Acceptable laboratory method blanks must not contain any
            signal at 320, 322, or 259 which is greater than 2% of the m/z
            332 response within +5 scans of the m/z 332 peak maximum.
            If the method blank that was extracted along with a batch of
            samples is contaminated, the associated positive samples must
            be rerun.  (See Exhibit C.)

            4.1.5.1  If the above criterion is not met, check solvents,
                     reagents, apparatus, and  glassware to locate and
                     eliminate the source of contamination before any
                     samples are extracted and analyzed.

            4.1.5.2  If new batches of reagents or solvents contain
                     Interfering contaminants, purify or discard them.

4.2'  Field Blanks Each batch of samples contains a sample of uncontami-
            nated soil/sediment and/or water that is to be fortified with
            unlabeled 2,3,7,8-TCDD at a concentration of 1 ug/kg for soil
            or 10 ng/L for water before analysis.  In addition to that
            field blank, a batch of samples may include a rinsate sample,
            that is a portion of solvent (usually trichloroethylene) that
            was used to rinse sampling equipment.  The rinsate is analyzed
            to assure that samples have not been contaminated by sampling
            equipment.

     4.2.1  Unfortified field blank  Analyze with procedures used for
            environmental samples (Section 11, Exhibit D).  This blank may
            or may not be labeled as such (i.e., it may be a "blind" QC
            sample).

     4.2.2  Fortified (Spiked) Field Blank                                晻

            4.2.2.1  Weigh a 10-g or measure a 1 L aliquot of the specified
                     .field blank sample and add 1.5 raL of th-> acetone
                     dilution of the 100 uL of field blank fortification
                     solution which contains 0.1 ng/uL of unlabeled
                     2,3,7,8-TCDD, 0.5 ng/uL of 13C12~2,3,7.8TCDD, and
                     0.014 ng/uL of 37Cl4-2,3,7,8-TCDD.
                                E - 3

-------
                 4.2.2.2  Extract with the appropriate method from Exhibit D
                          and analyze a 2梪L aliquot.

                 4.2.2.3  Calculate the concentration (Section 12.1, Exhibit
                          D) of unlabeled 2,3,7,8-TCDD, and the internal
                          standard recovery (Section 12.1.3, Exhibit D) of
                          the measured concentration.

          4.2.3  Rinsate Sample

                 4.2.3.1  To a 100-mL aliquot (or entire sample if less than
                          100 mL is provided) of equipment rinse solvent
                          (trichloroethylene-rinsate sample), add 1.5 mL of the
                          acetone dilution of 100 uL of the sample fortification
                          solution which contains 0.5 ng/uL of 13C12-2,3,7,8-TCDD
                          and 0.014 ng/uL of 37Cl4-2,3,7,8-TCDD.

                 4.2.3.2  Using a Kuderna-Danish apparatus, concentrate the
                          volume to approximately 5 raL.

                 4.2.3.3  Transfer the total 5-tnL concentrate in 1-mL portions to
                          a 1 mL-amber mini-vial, reducing volume as necessary
                          with a gentle stream of dry nitrogen.

                 4.2.3.4  Rinse container with two 0.5 mL portions of hexane and
                          transfer rinses to the 1-mL amber mini-vial.

                 4.2.3.5  Just before analysis, reduce volume to near dryness,
                          add 5 uL of the recovery standard solution and make
                          to a final volume of 50 uL with isooctane.  (Column
                          chromatography is not required.)

                 4.2.3.6  Analyze an aliquot with the same procedures used to
                          analyze samples (Section 11.5, Exhibit D).

5.   Duplicate Analyses

    5.1  Laboratory Duplicates  In each batch of samples, locate the sample
         specified for duplicate analyses and analyze a second sample aliquot.
         If no sample is specified for duplicate analysis the laboratory shall
         select one and analyze it in duplicate.  The sample chosen must not be
         the field blank.

         5.1.1  Results of laboratory duplicates must agree within  50% relative
                difference (difference expressed as percentage of the mean).,
                If the RPD is >50%, Contractor shall  immediately contact th<- '..'
                Sample Management Office for resolution of  the problem.  Report 
                all results.
                                      E  -  4

-------
                                 |SL - s2|  x 100   =  \BI  - s2|  x 100
                        RPD =  	     	
                               Mean Concentration         Sl + S2
                                                             2

                 Where S and S2 represent  sample and duplicate sample results.

          5.1.2  Recommended actions to help locate problem:

                 5.1.2.1  Analyze an aliquot of the performance check standard to
                          verify satisfactory instrument performance (Section 9,
                          Exhibit D).

                 5.1.2.2  If possible, determine that no error was made while
                          weighing or measuring sample aliquots.

                 5.1.2.3  Review analytical procedures with performing laboratory
                 "~        personnel.


6.   Identification criteria

     6.1  If any of the four initial identification criteria (Sections 11.6.1-
          11.6.4, Exhibit D) are not met, the sample is reported not to contain
          unlabeled 2,3,7,8-TCDD at the maximum possible concentration limit
          (Section 12.2, Exhibit D).

     6.2  When the four initial identification criteria are met,  but the fifth
          criteria, the isotopic abundance  ratio for m/z 320 and 322 (Section
          11.6.4, Exhibit D) is not met, that sample is presumed to contain
          interfering contaminants.  Contractor shall reextract,  clean-up, and
          reanalyze the sample.

     6.3  The recovery of the internal standard 13C12~2,3,7,8-TCDD should be
          within a 40 percent to 120 percent recovery window.  This is an
          advisory limit only, an action window may be set when sufficient data
          is available.

7.   Blind QC Samples  Included among soil and sediment or water samples may
     be QC samples that are not specified as such to the performing laboratory.
     Types that may be included are:

     7.1  Uncontaminated Soil or Water.

          7.1.1  If a false positive  is reported for this sample, the Contractor
                 shall be required  to rerun the entire associated batch of cauqles
                 (see Exhibit C).                                            .

     7.2  Split  Samples  composited sample aliquots sent  to more than one
          laboratory.

     7.3  Unlabeled Field Duplicates   two aliquots of a composited sample..

                                      E - 5

-------
     7.4  Performance Evaluation Sample  soil/sediment or water sample
          containing a known amount of unlabeled 2,3,7,8-TCDD.

          7.4.1  If the performance evaluation sample result falls outside the
                 acceptance windows established by EPA,  the Contractor shall be
                 required to rerun the entire associated batch of samples (see
                 Exhibit C).  NOTE:  EPA acceptance windows are based on
                 historical data results.
LABORATORY EVALUATION PROCEDURES

1.  On a continuing basis, the EPA Project Officer and/or designated represen-
    tatives may conduct an evaluation of the laboratory to ascertain that the
    'laboratory is meeting contract requirements.  This section outlines the
    procedures which may be used by the Project Officer or his authorized
    representative In order to conduct a successful evaluation of laboratories
    conducting dlbxin analyses according to this protocol.  The evaluation
    process consists of the following steps:  1) analysis of a performance
    evaluation (PE) sample, and 2) on-site evaluation of the laboratory to
    verify continuity of personnel, Instrumentation, and quality assurance/
    quality control functions.  The following Is a description of these two
    steps.

2.  Performance Evaluation Sample Analysis

    The PE samples are supplied by EMSL-LV to the EPA Regions who Include them
    with the cases submitted to the laboratories.  The PE samples are sent
    In this manner to assure that they are processed and reported In a routine
    manner by laboratory personnel.  The EPA Region client will evaluate the
    results to verify that the laboratory Is continuing to produce acceptable
    analytical results.  The acceptance windows provided by EMSL-LV are based
    on PE sample performance data and may be updated periodically as the size
    of the database increases.  The PE samples will be representative of the
    types of samples that will be subject to analysis under this contract.

3.   On-Slte Laboratory Evaluation

     3.1  An on-slte laboratory evaluation Is performed to verify that the
          laboratory Is maintaining the necessary minimum level in instru-
          mentation and levels of experience in personnel committed to the
          contract and that the necessary quality control/assurance activities
          are being carried out.  It also serves as a mechanism for discussing
          laboratory weaknesses identified through  routine data audits, PE
          sample analyses  results, and prior on-site evaluation.

     3.2  The sequence of  events  for the on-site evaluations  is shown  in
          Figure  1.  The  Site  Evaluation Sheet  (SES)  (Figure  2) is used to
          document the results of  the evaluation.
                                      E  -  6

-------
                              Figure 1                             EXAMPLE

                  EVENT SEQUENCE FOR  SITE  EVALUATION


I.  MEETING WITH LABORATORY MANAGER AND PROJECT MANAGER

General discussion of purpose of site visit, purpose of analyses and current
contract award status.

II.  VERIFICATION OF PERSONNEL

Review qualifications of contractor personnel in place and committed to
project (Section I, SES).

III.  VERIFICATION OF INSTRUMENTATION

Review equipment in place and committed to project (Section II, SES).  The
bidder must demonstrate adequate equipment redundancy to ensure capability to
perform required analyses in the required time.

IV.  QUALITY CONTROL PROCEDURES

    Walk through laboratory to review:

    1.   Sample reception and logging procedures
    2.   Sample and extract storage area,
    3.   Procedures to prevent sample contamination,
    A.   Security procedures for laboratory and samples,
    5.   Safety procedures,
    6.   Conforaance to written Standard Operating Procedures,
    7.   Instrument records and logbooks,
    8.   Sample and data control systems,
    9.   Procedures for handling and disposing of hazardous materials,
    10.  Glassware cleaning procedures,
    11.  Status of.equipment and its availability,
    12.  Procedures for data handling, analysis, reporting, and case file
'_        preparation and
:   13.  Chain-of-custody procedures.

V.  REVIEW OF STANDARD OPERATING PROCEDURES (SOPs)

Review SOPs with project manager to assure that the laboratory understands
the dimensions and requirements of this program.                          ;   

VI.  IDENTIFICATION OF NEEDED CORRECTIVE ACTIONS                          '

Discuss with project manager, the actions needed to correct weaknesses identified
during site Inspection, PE sample analysis or production'of reports (hard'
copies and magnetic tapes) and documentation.  Determine how and when corrective
actions will be documented, how and when improvements will be demonstrated, and
the contractor employee responsible for corrective actions.
                                     E-7

-------
                             .    FiRure 2                          EXAMPLE



                     LABORATORY SITE EVALUATION SHEET (SES)
Laboratory:




Date:
Type of Evaluation:




Contract Number:




Contract Title:
Personnel Contacted:
                 Name                                      Title
Laboratory Evaluation Team:
                 Name                                      Title
                                          E-8

-------
I.   Organization and Personnel  (Page 1 of 2)
                                                               EXAMPLE
ITEM
Laboratory or Project Manager (individual
responsible for overall technical effort):
Name :

GC/MS Operator:
Name:
Experience: 1 year minimum requirement per
appropriate instrument
GC/MS Spectral Interpretation Expert:
Name:
Experience: 2 years minimum requirement
Extraction Concentration Expert:
Name:
Experience: 6 months minimum requirement
Do personnel assigned to this project have the
appropriate educational background to success-
fully accomplish the objectives of the program?
Do personnel assigned to this project have the
appropriate level and type of experience to
successfully accomplish the objectives of this
program?
Is the organization adequately staffed to
meet project commitments in a timely manner?
YES







NO







COMMENT







                                      E-9

-------
                                                               EXAMPLE
I.   Organization and Personnel  (Page 2 of 2)
ITEM
Does the laboratory Quality Assurance
Supervisor report to senior management levels?
Was the Project Manager available during the
evaluation?
Was the Quality Assurance Supervisor available
during the evaluation?
YES



NO



COMMENT



                                  E-LO

-------
A. General Facilities (Page .1 of 6)
                                                                 EXAMPLE
Does Che laboratory appear to have adequate
workspace (120 sq. feet, 6 linear feet of
unencumbered bench space per analyist)?
Are voltage control devices used on major
instrumentation?
Does the laboratory have a source of distilled/
demineralized water?
I,s the conductivity of distilled/demineralized
water routinely checked and recorded?
Is the analytical balance located away from
draft and areas subject to rapid temperature
changes?
Has the balance been calibrated within one year
by a certified technician?
Is the balance routinely checked with class S
weights before each use and the results recorded
in a logbook?





















                                  E-ll

-------
                                                          EXAMPLE
A.  General Factltties  (Page 2 of 6)
ITEM
Are properly filtered exhaust hoods provided to
allow efficient work with hazardous/toxic
materials?
Is the laboratory maintained In a clean and
organized maner?
Is a glove box available to allow efficient
work with hazardous/toxic materials?
Are contamination-free work areas provided for
the handling of toxic materials?
Are the toxic chemical handling areas either a
stainless steel bench or an impervious material
covered with absorbent material?
Are adequate facilities provided for storage of
samples, extracts, and calibration standards,
including temperature controlled storage?
Is the temperature of the cold storage units
recorded daily in logbooks?
Are chemical waste disposal policies/procedures
adequate?
Are contamination-free areas provided for trace
level analytical work?
Can the laboratory supervisor document that
trace-free water is available for preparation
of standards and blanks?
YES










NO










COMMENT








'

                                  E-12

-------
                                                          EXAMPLE
A.  General Facilities  (Page 3 of 6)
ITEM
Is the laboratory secure?
Can the laboratory supervisor document that
organic solvents used are free of trace
contaminants?
YES


NO


COMMENT


Comments on Laboratory Facilities
                                     E-13

-------
                                                          EXAMPLE
B.  Equipment  (Page 4 of 6)




    1.  GC/MS/DS Instrumentation







                   Manufacturer
Model
Installation Date
HRGC/MS HRMS
ID it
GC/MS
ID ft
Peak Matching
Unit ID 0
GC (interfaced
with MS) ID#
Data System
IDtf
Data System
ID #


















Comments on GC/MS/DS Instrumentation:
                                   E-14

-------
                                                           EXAMPLE
B.  Equipment  (Page 5 of 6)
ITEM
Are manufacturer's operating manuals readily
available to the operator?
Is there a calibration protocol available to
the operator?
Are calibration results kept in a permanent
record?
Is service maintenance by contract?
Is preventative maintenance applied?
Is a permanent service record maintained in a
logbook?
Has the instrument been modified in any way?
Is the instrument properly vented?
Is a 9-track mag- tape available?
Is a split/splitless capillary injector
in place?
Is the column direct to the source?
Are sufficient in-house replacement parts
available?
YES












NO












COMMENT












                                   E-15

-------
                                                          EXAMPLE




B.  Equipment  (Page 6 of 6)



Comments on GC/MS Instrumentation
                                  E-16

-------
                                                             EXAMPLE
III.  Documentation  (Page 1 of 2)

      When reviewing documentation, give special attention to:

           a) traceabllity
           b) neatness and completion

A.  Documentation/Tracking
ITEM
Is a sample custodian designated? If yes,
name of sample custodian.
Name:

Are the sample custodian's procedures and
responsibilities documented? If yes, where
are these documented?
Are written Standard Operating Procedures (SOP)
developed for receipt of samples? If yes,
where are the SOP documented (laboratory manual,
written instructions, etc.)?
Are quality assurance procedures documented
and available to the analysts? If yes, where
are these documented?
Are written Standard Operating Procedures (SOP)
developed for compiling and maintaining sample
document files? If yes, where are the SOP
documented (laboratory manual, written
instructions, etc.)?
Are the magnetic tapes stored in a secure area?
Is a permanently bound notebook with preprinted,
consecutively numbered pages being used?
YES







NO







COMMENT





-

                                    E-17

-------
                                                          EXAMPLE
B.  Documentation/Notebooks  (Page 2 of 2)
ITEM
Is the type of work clearly displayed on the
notebook (i.e., EPA Extraction)?
Is the notebook maintained In a legible manner?
Are entries noting anomalies routinely
Recorded?
Has the analyst avoided obliterating entries?
Are Inserts (I.e., chromatograms, computer
printout, etc.) permanently affixed in notebook
and signed across insert edge and page?
Has the supervisor of the individual maintaining
the notebook personally examined and reviewed
the notebook periodically, and signed his/her
name therein, together with the date and appro-
priate comments as to whether or not the
notebook is being maintained in an appropriate
manner?
Where applicable, is the notebook holder
referencing reports or memoranda pertinent
to the contents of an entry?
YES







NO







COMMENT







                                  E-18

-------
                                                           EXAMPLE
IV.   Analytical Methodology  (Page 1 of 2)
ITEM
Are the required methods used?
Is there any unauthorized deviation from
contract methodology?
Are written analytical procedures provided to
the analyst?
Are distilled-in-glass grade or other high
purity chemicals used to prepare standards?
Are fresh analytical standards prepared at a
frequency consistent with good QA?
Are reference materials properly labeled with
concentrations, date of preparation, and the
identity of the person preparing the sample?
Is a standards preparation and tracking logbook
maintained?
Do the analysts record bench data in a neat and
accurate manner?
Is the appropriate instrumentation used in .
accordance with the required protocol(s)?
YES









NO









COMMENT









                                  E-19

-------
                                                          EXAMPLE





Comments on Analytical Methods and Practices  (Page 2 of 2)
                                  E-20

-------
                                                             EXAMPLE
V.  Quality Control Manual Checklist  (Page 1 of 2)
ITEM
Does the laboratory maintain a Quality Control
Manual?
Does the manual address the important elements
of a QC program, including the following:
a. Personnel?
b. Facilities and equipment?
c. Operation of instruments?
d. Documentation of procedures?
e. Procurement and inventory practices?
f. Preventive maintenance?
g. Reliability of data?
h. Data validation?
i. Feedback and corrective action?
j. Instrument calibration?
k. Recordkeeping?
1. Internal audits?
YES














NO














COMMENT










.



                                     E-21

-------
                                                         EXAMPLE
V.  Quality Control Manual Checklist  (Page 2 of 2)
ITEM
Are QC responsibilities and reporting relation-
ships clearly defined?
Have standard curves been adequately documented?
Are laboratory standards traceable?
Are quality control charts maintained for each
routine analysis?
Do QC records show corrective action when
analytical results fail to meet QC criteria?
Do supervisory personnel review the data and
QC results?
YES






NO






COMMENT






                                 E-22

-------
VII.  Summary




A.  Summary Checksheet (Page 1 of 2)
                                                        EXAMPLE
ITEM
Do responses to Che evaluation Indicate that
project and supervisory personnel are aware of
QA and Its application to the project?
Do project and supervisory personnel place
positive emphasis on QA/QC?
Have responses with respect to QA/QC aspects of
the project been open and direct?
Has a cooperative attitude been displayed by all
project and. supervisory personnel?
Does the organization place the proper emphasis
on quality assurance?
Have any QA/QC deficiencies been discussed
before leaving?
Is the overall quality assurance adequate to
accomplish the objectives of the project?
Have corrective actions recommended during
previous evaluations been implemented?
Are any corrective actions required? If so,
list the necessary actions below.
YES









NO









COMMENT









                                 E-24

-------
                                                        EXAMPLE
VI.  Data Handling Checklist  (Page 1 of 1)
ITEM
Are data calculations checked by a second
person?
Are data calculations documented?
Do records indicate corrective action that has
been taken on rejected data?
Are limits of detection determined and reported
properly?
Are all data and records retained for the
required amount of time?
Are quality control data (e.g. , standard curve,
results of duplication and spikes) accessible
for all analytical results?
YES






NO






COMMENT






                                 E-23

-------
                                                          EXAMPLE



B.   Summary Comments and Corrective Actions  (Page 2 of 2)
                                     E-25

-------
                          EXHIBIT  F
                     SPECIFICATIONS FOR
             CHAIN-OF-CUSTODY,  DOCUMENT  CONTROL,
              AND STANDARD OPERATING  PROCEDURES
NOTE:  The Contractor shall not deviate from the procedures
       described herein without the prior written approval
       of the Contracting Officer:  Provided, that the
       Contracting Officer may ratify in writing such
       deviation and such ratification shall constitute the
       approval required herein.

-------
     SPECIFICATIONS  FOR CHAIN-OF-CUSTODY,  DOCUMENT  CONTROL
         PROCEDURES,  AND STANDARD  OPERATING  PROCEDURES
1.   SAMPLE CHAIN-OF-CUSTODY

          A sample is physical evidence collected from a
     facility or from the environment.   An essential part of
     the hazardous, waste investigation  effort is that the
     evidence gathered be controlled.  To accomplish this, the
     following chain-of-custody procedures have been
     established.

     1.1  Sample Identification

          To assure the traceability of samples through the
          -laboratory, a method for sample identification shall
          be developed and documented in the laboratory SOPs
          (see Section 3).  Each sample or sample preparation
          container shall be labelled with a unique number
          identifier (or the EPA sample number). This identifier
          shall be cross-referenced to  the sample tag number
          and the EPA sample number.  There shall be a written
          description of the method of  assigning this
          identifier and attaching it to the sample bottle,
          included in the laboratory SOPs.

     1.2  Chain-of-Custody Procedures

          Because of the nature of the  data being collected,
          the possession of samples must be traceable from the
          time the samples are collected until they are
          introduced as evidence in legal proceedings.  To
          maintain and document sample custody, the chain-of-
          custody procedures described below shall be
          followed.

          1.2.1  A sample is under custody if:

                 1.2.1.1  It is in your actual possession,

                 1.2.1.2  It is in your view after being in
                          your physical possession,

                 1.2.1.3  It was in your possession and then
                          you locked or sealed it up to
                          prevent tampering, or

                 1.2.1.4  It is in a secure area.

          1.2.2  Upon receipt of the samples in custody, the
                 contractor shall inspect the shipping
                 container and sample bottles, and shall
                 document receiving information as specified


                          F-l

-------
in Section 3.2.  The sample custodian or a
designated representative shall sign and date
all appropriate receiving documents at the
time of receipt (i.e., EPA chain-of-custody
forms, traffic reports, airbills, etc.).  The
contractor shall contact SMO if documents are
absent, information on receiving documents
does not agree, custody seals are not intact,
or the sample is not in good condition.  The
contractor shall document resolution of any
discrepancies.
         F-2

-------
2.   DOCUMENT CONTROL  PROCEDURES

          The goal  of the  laboratory document control program
    is to assure  that all  documents for a specified case will
    be accounted  for  when  the project is completed.
    Accountable documents  used by contract laboratories shall
    include,  but  not  be  limited to, logbooks, chain-of-
    custody records,  sample work sheets, bench sheets, and
    other documents relating to the sample or sample
    analyses.   The  following document control procedures have
    been established  to  assure that all laboratory records
    are assembled and stored for delivery to EPA or are
    available upon  request form EPA prior to the delivery
    schedule.

    2.1  Preprinted Data Sheets and Logbooks

         Preprinted data sheets shall contain the name of the
         laboratory and  be dated and signed by the analyst  or
         individual performing the work.  All documents
         produced by  the laboratory which are directly
         related  to the  preparation and analysis of EPA
         samples  shall become the property of the EPA and
         shall be placed in the case file.  For that  reason,
         all observations  and results recorded by the
         laboratory but  not on preprinted data sheets are
         entered  into permanent laboratorey logbooks.  The
         person responsible for the work shall sign and date
         each entry and/or page in the logbook.  When all
         data from  a  case  is compiled, copies of all  EPA
         case-related logbook entries shall be included in
         the documentation package.  Analysts' logbook
         entries  must be in chronological order and shall
         include  only one  case per page.  Instrument  run  logs
         shall be maintained so as to enable a reconstruction
         of the run sequences of individual instruments.

         Because  the  laboratory must provide copies of the
         instrument run  logs to EPA, the laboratory may
         exercise the option of using only laboratory or  SMO
         sample identification numbers in the logs  for  sample
         ID rather  than  government agency or commercial
         client names.

         Using laboratory  or SMO sample  IDs only  in  the  run
         sequences  will  assist the  laboratory  in preserving
         the confidentiality of commercial clients,

    2.2  Error Correction  Procedure

         All documentation in  logbooks  and  other  documents
         shall be in  ink.   If  an  error  is made"7 corrections
         shall be made by  crossing  a line  through the error
                       F-3

-------
     and entering the correct information.   Changes shall
     be dated and initialed.  No information shall be
     obliterated or rendered unreadable.

2.3  Consistency of Documentation

     Before releasing analytical results, the laboratory
     shall assemble and cross-check the information on
     sample tags, custody records, lab bench sheets,
     personal and instrument logs, and other relevant
     data to ensure that data pertaining to each
     particular sample or case is consistent throughout
     the case file.

2.4  Document Numbering and Inventory Procedure

    桰n order to provide document accountability of the
     completed analysis records, each item in a case
     shall be inventoried and assigned a serialized
     number and identifier associating it to the case and
     Region.

     Case # - Region - Serialized number (For example:
     75-2-0240)

     The number of pages of each item must be accounted
     for if each page is not individually numbered.  All
     documents relevant to each case, including logbook
     pages,  bench sheets, mass spectra, chromatograms,
     custody records, library search results, etc., shall
     be inventoried.  The laboratory shall be responsible
     for ensuring that all documents generated are placed
     in the file for inventory and are delivered to EPA.
     Figure 1 is an example of a document inventory.

2.5  Shipping Data Packages and Case Files

     The contractor shall have written procedures to
     document shipment of deliverables packages to the
     recipients.  These shipments require custody seals
     on the containers placed such that it cannot be
     opened without damaging or breaking the seal.  The
     contractor shall also document what was sent, to
     whom,  the date, and the method (carrier) used.
                   F-4

-------
3.  SPECIFICATIONS  FOR_ANPARD OPERATING PROCEDURES

          The contractor must have written standard operating
     procedures  (SOPs) for receipt of samples, maintenance of
     custody,  sample storage, tracking the analysis of samples
     and assembly of completed data.

          An  SOP is defined as a written narrative step-wise
     description of laboratory operating procedures including
     examples  of laboratory documentation.  The SOPs must
     accurately  describe the actual procedures used in the
     laboratory, and copies of the written SOPs shall be
     available to the appropriate laboratory personnel.  These
     procedures  are necessary to ensure that analytical data
     produced  under this contract are acceptable for use in
     EPA enforcement case preparation and litigation.  The
     contractor's SOPs shall provide mechanisms and
     documentation to meet each of the following
     specifications and shall be used by EPA as the basis for
     laboratory  evidence audits.

     3.1  The contractor shall have a designated sample
         custodian responsible for receipt of samples and
         have written SOPs describing his/her duties and
         responsibilities.

    3.2  The contractor shall have written SOPs for receiving
         and logging in of the samples.   The procedures shall
         include but not be limited  to documenting the
         following information:

         3.2.1  Presence or absence  of EPA chain-of-custody
                forms

         3.2.2  Presence or absence  of airbills

         3.2.3  Presence or absence  of traffic reports or SAS
                packing  lists

         3.2.4  Presence or absence  of custody seals on
                shipping and/or sample containers and their
                condition

         3.2.5  Presence or absence  of sample tags

         3.2.6  Sample tag  ID  numbers  if  not recorded on the
                chain-of-custody  record(s)  or packing list(s)

         3.2.7  Condition of the  shipping container

         3.2.8   Condition of the  sample bottles

        3.2.9   Verification of agreement or non-agreemerjt of.
                information on receiving  documents
                       F-5

-------
     3.2.10 Resolution of problems or discrepancies with
            the Sample Management Office

 3.3  The contractor shall have written SOPs for
     maintenance of the security of samples after log-in
     and shall demonstrate, security of the sample storage
     areas and laboratory.  The SOPs shall specifically
     include .descriptions of all storage areas for EPA
     samples in the laboratory.  -The SOPs shall include a
     list of authorized personnel who have access or keys
     to secure storage areas.

 3.4  The contractor shall have written SOPs for tracking
     the work performed on any particular sample.  The
     tracking SOP shall include the following:

     3.4.1  A description of the documentation used to
            record sample receipt, sample storage, sample
            transfers, sample preparations, and sample
            analyses.

     3.4.2  A description of the documentation used to
            record calibration and QA/QC laboratory work.

     3.4.3  Examples of the document formats and
            laboratory documentation used in the sample
            receipt,  sample storage, sample transfer, and
            sample analyses.
                                         //

3.5  The contractor shall have written SOPs for
     organization and assembly of all documents relating
     to each EPA case.  Documents shall be filed on a
     case-specific basis.  The procedures must ensure
     that all documents including logbook pages, sample
     tracking records, chromatographic charts, computer
     printouts,  raw data summaries, correspondence, and
     any other written documents having reference to the
     case are compiled in one location for submission to
     EPA.   The system must include a document numbering
     and inventory procedure.

-------
HANDLING OF CONFIDENTIAL INFORMATION

     A contractor conducting work under this contract: may
receive EPA-designated confidential information from the
agency.  Confidential information must be handled
separately from other documentation developed under this
contract.  To accomplish this, the following procedures
for the handling of confidential information have been
established.

4.1  All confidential documents shall be under the
     supervision of a designated document control officer
     (DCO).

4.2  Confidential Information

    -Any samples or information received with a request
     of confidentiality shall be handled as
     "confidential."  A separate locked file shall be
     maintained to store this information and shall be
     segregated froi*. other nonconfidential information.
     Data generated from confidential samples shall be
     treated as confidential.  Upon r^r-eipt of
     confidential irformatinr,  wt>.? JCO lo^t- these
     documents ijaf* a'-Jbnfidential  Inventory Locr.   The
     informx-Ion is then made available to autho.-ized
     personnel but only after it has been signed out: to
     the person by the DCO.   The documents shall be
     r^urr.ed to the  locked file at the conclusion of
     -昦ch working day.  Confidential information may not
     be reproduced except upon approval by the EPA
     contracting officer.   The DCO  will enter all copies
     into the document control system.  In addition, this
     information may  not be disposed of except upon
     approval by the  EPA contracting officer.  The DCO
     shall remove and retain the cover page of any
     confidential information disposed of for one year
     and shall keep a record of the disposition in the
     Confidential Inventory Log.
                    F-7

-------
                              Figure 1
                                             232-2-0001

                                             Case No. 232
                               Example
Document Control #*

232-2-0001
232-2-0002
232-2-0003   -*--
232-2-0004
232-2-0005
232-2-0006
232-2-0007
232-2-0008
232-2-0009
232-2-0010
232-2-0011
232-2-0012
   etc.
     DOCUMENT INVENTORY

       Document Type

Case File Document Inventory Sheet
Chain-of-Custody Records
Shipping Manifests
Sample Tags
SMO Organics Traffic Reports
GC/MS spectra for sample B0310
GC/MS spectra for sample B0r*ll
GC/MS spectra for sample BO319
Analyst's logbook pages
GC/MS library search worksheets
GC instrument log pages
GC/MS QC data sheets
        etc.
# Pages

   1
   2
   2
  50
  10
  20
  20
  20
   6
  15
   5
   4
 etc.
* This number is to be recorded on each  set  of  documents.
                           F-8

-------