vvEPA
           United States
           Environmental Protection
           Agency
           Environmental Monitoring
           Systems Laboratory
           P.O. Box 15027
           Las Vegas NV 89114-5027
EPA 600/4-86-004
January 1986
           Research and Development
Protocol for the
Analysis of 2,3,7,8-
Tetrachlorodibenzo-p-Dioxin
by High-Resolution Gas
Chromatography/
High-Resolution
Mass Spectrometry

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        PROTOCOL FOR THE ANALYSIS OF 2,3,7,8-TETRACHLORODIBENZO-£-DIOXIN BY
        HIGH-RESOLUTION GAS CHROMATOGRAPHY/HIGH-RESOLUTION MASS SPECTROMETRY
                                         by

                         John S.  Stanley and Thomas M.  Sack
                             Midwest Research Institute
                            Kansas City, Missouri  64110
                              Contract Number SAS 1576X
I
                                   Project Officer

                                  Werner F. Beckert
                             Quality Assurance Division
                     Environmental Monitoring Systems Laboratory
                              Las Vegas, Nevada  89114
                     ENVIRONMENTAL MONITORING SYSTEMS LABORATORY
                         Office of Research and Development
                        U.S. Environmental Protection Agency
                              Las Vegas, Nevada  89114

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                                   NOTICE
     The information in  this  document has been funded wholly  or  in part by
the  United States  Environmental Protection  Agency  under  Contract  Number
SAS 1576X to the Midwest Research Institute,  Kansas City, Missouri.   It has
been subject to the Agency's peer and administrative review,  and it has  been
approved  for  publication  as  an  Environmental  Protection Agency  document.
Mention of  trade names  or commercial products does not  constitute endorse-
ment or recommendation for use.

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                                   PREFACE
     This  report  describes the  activities  completed as  part of a  single-
laboratory   evaluation  of   a  high-resolution   gas   chromatography/high-
resolution mass  spectrometry method  for the determination  of tetrachloro-
dibenzo-£-dioxins in  water,  soil,  and sediment samples.  The work described
in  this  report was  completed at the Midwest Research  Institute  under con-
tract  to  Viar and  Company (Special Analytical Services  SAS  1576X)  for the
U.S.  Environmental  Protection  Agency,  Environmental  Monitoring  Systems
Laboratory, Quality Assurance Division,  Las Vegas, Nevada.   The revision of
the protocol  to  allow for lower quantitation limits for tetrachlorodibenzo-
£-dioxins was carried out at the Environmental Monitoring Systems Laboratory-
Las Vegas.

     This  report was  prepared with assistance from M.  McGrath.  The authors
acknowledge the technical  project  monitor,  W. F. Beckert, as  well  as  R. K.
Hitchum and  S. Billets of the Environmental  Monitoring Systems Laboratory-
Las Vegas  and, especially, Y. Tondeur of the Environmental Research Center,
University of Nevada, Las Vegas for guidance  provided  during  this  study.
                                     111

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                                  ABSTRACT
     This report provides the results of the single-laboratory evaluation of
a  high-resolution   gas   chromatography/high-resolution  mass  spectrometry
method  for  the  determination of  2,3,7,8-tetrachlorodibenzo-£-dioxin  and
total  tetrachlorodibenzo-£-dioxins  at concentrations  ranging  from 10  to
200 pg/g  (ppt)  in soils  and  100  to 2,000 pg/L (ppq) in  water.   The report
summarizes  the  data for  the  precision and accuracy  of triplicate measure-
ments of five solid and five aqueous samples.   The results indicate that the
method is capable of generating accurate and precise data within the concen-
tration limits  specified  above and within absolute recoveries  of 40 to 120
percent with 50 percent precision.  An attempt to reach a quantitation limit
for TCDD  of 2 ppt (or less) for  soil  and 20  ppq (or less) for aqueous sam-
ples was  not successful.   Based on the data generated during this study and
based  on  discussions  at  the  Environmental Monitoring  Systems  Laboratory-
Las Vegas, the Environmental Monitoring Systems Laboratory-Las Vegas revised
certain parts of the  protocol to  lower  the  quantitation  limit for tetra-
chlorodibenzo-£-dioxins to 2 ppt in soil and 20 ppq in water samples.
                                     IV

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                                  CONTENTS
Preface	    iii
Abstract	     iv
Figures	     vi
Tables	    vii

   1.  Introduction 	      1
   2.  Conclusions	      3
   3.  Recommendations	      5
   4.  Experimental Procedures	      7
            Sample description	      7
            Sample preparation	      7
            Reagents	      9
            HRGC/HRMS instrumentation 	      9
            Mass measurement accuracy 	     11
            Chromatographic resolution	     13
            Injection technique 	     13
   5.  Results and Discussion	     14
            Approach to cleanup column evaluation 	     14
            Final method evaluation 	     22

References	     42
Appendices	     43
   A.  Validated Analytical Protocol
   B.  Proposed Analytical Protocol

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                                   FIGURES

Number                                                                  Page

  1    Column cleanup procedures specified in the protocol	      15

  2    Column cleanup procedures proposed by the EMSL-LV	      16

  3    Background levels of 1,3,6,8- and 1,3,7,9-TCDD observed over
         the single-laboratory evaluation study 	      41
                                     VI

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                                   TABLES

Table                                                                   Page

  1    Solid Samples Used for HRGC/HRMS Method Evaluation 	       8

  2    Aqueous Samples Used for HRGC/HRMS Method Evaluation 	       8

  3    TCDD Isomers Used for HRGC/HRMS Method Evaluation	      10

  4    Composition of Concentration Calibration Solutions (pg/pL) .  .      10

  5    HRGC/HRMS Operating Conditions 	      12

  6    Recovery (%) of Several TCDD Isomers from Cleanup Option A .  .      18

  7    Recovery (%) of Several TCDD Isomers from Cleanup Option B .  .      19

  8    Recovery (%) of Several TCDD Isomers from Cleanup Option C .  .      20

  9    Recovery (%) of Several TCDD Isomers from Cleanup Option D .  .      21

 10    Initial Calibration Summary	      23

 11    HRGC and Mass Resolution Check Summary	      24

 12    TCDD Data Report Form	      26

 13    Accuracy and Precision of the HRGC/HRMS Analysis for
         2,3,7,8-TCDD from Laboratory Aqueous Matrix Spikes 	      33

 14    Precision of the HRGC/HRMS Analysis for 2,3,7,8-TCDD of
         Soil and Fly Ash Samples	      34

 15    Accuracy of the HRGC/HRMS Method for the Determination of
         TCDD Isomers Spiked into Aqueous Matrices	      35

 16    Accuracy of the HRGC/HRMS Method for the Determination of
         TCDD Isomers Spiked into Soil Matrices	      36

 17    Fortified Field Blank Results	      37
                                     VI1

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                                  SECTION 1

                                INTRODUCTION

     The  U.S.  Environmental Protection Agency's  (EPA)  strategy  for dealing
with dioxin  requires  the development and validation of an analytical method
capable  of  achieving detection of  the tetrachlorodibenzo-£-dioxins (TCDD),
specifically 2,3,7,8-TCDD, at the parts-per-trillion (ppt) level in soil and
sediment  and parts-per-quadrillion  (ppq)  level  in water.1   This validated
method will be used by qualified contract laboratories to extend the analyt-
ical capabilities for such analyses to all EPA regional and program offices.

     This report deals specifically with the single-laboratory evaluation of
a  high-resolution  gas   chromatography/high-resolution  mass  spectrometry
(HRGC/HRMS)  analysis  method for  TCDDs in  soil,  sediment, and  water.   The
method  (Appendix A)   is  intended to provide quantitative  determination of
TCDD at  levels  of 10 to 200 pg/g  (soil  and sediment) and 100 to 2,000 pg/L
(water)  at  a mass resolution of  10,000.   This  single-laboratory evaluation
has been completed as  part of the validation process  recommended by EPA.2

     The  proposed method was prepared after several  candidate  methods were
reviewed and their best features were selected.   After peer review, the pro-
posed method was refined for completeness, technical accuracy,  clarity, and
regulatory applicability.   The  single-laboratory evaluation of the proposed
analytical method has been accomplished through three tasks.  The first task
involved  preliminary  performance  testing  of   the   method  using  TCDD-
contaminated  soils and  TCDD-spiked aqueous samples.   The results  of this
study  indicated  that the  proposed  method  required modification to achieve
the target method detection limits and the accuracy and precision criteria.
The second task  focused on ruggedness testing of the chromatographic cleanup
procedures.   The results  of this  study were used  to modify  the proposed
method.   This  report is  focused on the results  of the triplicate analysis
of five  solid and  five aqueous  samples  completed under  the third task of
the evaluation,  using the modified method.

     Section  2  of this   report  summarizes the  conclusions  based  on the
single-laboratory  evaluation of  this  method using TCDD-contaminated soils
and TCDD-spiked  aqueous samples.   Section 3 presents  recommendations that
should  be considered  for  inclusion in  the method before proceeding with
collaborative  testing.   Section 4 presents  some  specific experimental con-
ditions,  and Section 5  summarizes  the  analytical data  for  the triplicate
analysis  of  four soil,  one fly ash, and five aqueous  samples  completed in
the third  task of the single-laboratory evaluation.  Triplicate analyses of
a  1-pg/pL  calibration solution did not give satisfactory results.  In order
to achieve a quantitation limit of 2 ppt for soil (using a 10-g sample) and

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20 ppq  for water  (using a  2.0-L  sample), the  protocol  evaluated  in  this
study was  modified.   The  rationale  for the  modifications  and  the  revised
protocol are included as Appendix B.

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                                  SECTION 2

                                 CONCLUSIONS

     The  single-laboratory  evaluation of  the  analytical  method for  the
determination of  2,3,7,8-TCDD  in soil and aqueous samples demonstrates that
the method  as  described is capable of achieving the target detection limits
of 10 pg/g  (ppt) for soils and 100 pg/L (ppq) for water.

     The relative  response factors  (RRF)  determined for native 2,3,7,8-TCDD
versus the internal standard l3C12-2,3,7,8-TCDD, and the RRF of the internal
standard versus the recovery standard 13C12~1|2,3,4-TCDD over the five-point
concentration calibration  curve  demonstrate that the HRGC/HRMS method main-
tains a  linear response  for  2,3,7,8-TCDD from 10 to 200 ppt  for  soils  and
100 to 2,000 ppq for water.

     The results  of  the analysis of spiked aqueous samples demonstrate that
internal standard  (isotope dilution)  quantitation provides an accurate mea-
surement of  2,3,7,8-TCDD.   The accuracy of the 2,3,7,8-TCDD measurement for
triplicate  analysis  of four water samples  spiked  at various concentrations
was quite good.   The accuracy of measurement  for  2,3,7,8-TCDD averaged  104
percent  for three  aqueous matrices  prepared  as laboratory matrix  spikes.
The absolute recovery  of  the  internal standard  13C!2-2,3,7,8-TCDD  did  not
significantly affect the  accuracy  of the  2,3,7,8-TCDD determination.   The
precision of the analyses  for 2,3,7,8-TCDD ranged from 3.6 to 16 percent for
replicate analyses of  the five aqueous samples.  The precision of the trip-
licate analyses of the soil samples was somewhat higher than determined for
aqueous  samples.   The  precision of  triplicate analyses  of the  four  soil
samples ranged from 19  to  50 percent.  The difference in precision from that
of the aqueous samples may be attributable to the potential for TCDD adsorp-
tion on the soil samples.

     The results  from  the analyses  of soil and  aqueous samples spiked with
additional TCDD isomers demonstrate that  the internal standard quantitation
gives good  estimates  of total TCDD values.  The accuracy of the analyses of
fortified distilled  water and  influent  and effluent  wastewaters averaged
101 ± 14 percent for five  TCDD isomers.  The accuracy of the measurements of
these isomers for the  four fortified soil samples averaged 87 t 24 percent.

     The results of the analyses demonstrate that the requirements for abso-
lute recovery of  the internal standard (40 to 120 percent) and precision of
replicate analyses  (RPD < 50 percent) can be  achieved  for  relatively clean
samples.

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     The sample matrix can severely impact the performance of the analytical
method.  This  is evidenced  by the consistent low recovery  of  the internal
standard from the fifth aqueous sample, an industrial wastewater, and from a
fly ash sample.  The low recovery from the industrial wastewater is possibly
due  to the effect  of coextractants  on  the elution  sequence  from alumina.
The  low  recoveries  observed for the  fly  ash  sample,  on the other hand, may
be attributed to adsorption by the sample matrix.

     One of the most critical variables in the analytical method is the com-
pleteness of removal  of the benzene from the extract before proceeding with
the  acidic alumina  column  fractionation.   The  cleanup column  ruggedness
testing experiments demonstrated that the recoveries of 2,3,7,8-TCDD and the
other  TCDDs  are affected by the  presence of benzene in the alumina column
fractionation step.

     The analyst  must be  aware of the  potential problem of  interferences
arising  from  background  contamination.   For  example, the  1,3,6,8-  and
1,3,7,9-TCDD  isomers  were  present in  the fortified  field  blanks  in this
work.  From other  referenced activities  it becomes clear that these isomers
may  present  problems  in  other laboratories as  well.    The  fortified  field
blanks are  important  tools in assessing  the  background contamination  prob-
lems over time.

     Although  the  1.0-pg/pL standard did not yield  satisfactory results in
this study, due  to unacceptable ion ratios, the response factors are within
the established curve.  The data for the triplicate analyses of the 1.0-|Jg/pL
standard demonstrate that the characteristic ions for TCDD were greater than
20:1 for the m/z 322  S/N and approximately 10:1  for  m/z 259 S/N.   Thus, it
should be possible  to extend the detection limit to 1 pg/pL if an allowance
for abundance ratios based on ion statistical errors is  incorporated.

     Based on  the column performance and bleed  characteristics,  the column
of choice for the analysis for TCDD at ppt (for soils and sediments) and ppq
(for  water)  levels  appears  to  be the  50-m  CP-Sil 88 with a  0.2-pm film
thickness.   To preserve the performance characteristics  of the HRGC columns,
an injection technique that excludes any air is highly recommended.

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                                 SECTION 3

                              RECOMMENDATIONS

1.  Mass measurement accuracy should properly be determined relative to the
    lock mass  (if  any),  rather than m/z 254.9856, because it is that rela-
    tionship which will  determine  how  accurately the  masses  of the  TCDD
    ions will be measured.

2.  It  is  recommended that  the chromatographic  resolution check  be  per*
    formed on  the  summed ion chromatograms of m/z 259 + m/z 320 + m/z 322.
    This yields a  chromatogram which is less noisy and more representative
    of the true column performance.

3.  The 5  percent peak  width criterion for mass resolution should be the
    selected  mass/1,000   mmu  rather  than  31.9  nunu  because the  protocol
    allows peaks other than  m/z 319 to be  used  for resolution measurement
    (e.g., 31.7 mmu if m/z 317 is used).

4.  It  is  recommended that  the mass measurement accuracy  be  recorded and
    reported along with the resolution check summary table.

5.  The  addition  of  the  recovery standard  13C!2-1,2,3,4-TCDD should  be
    achieved by  using a  spike volume  of  25 to 50 pL  rather  than  5 |JL to
    minimize errors resulting from volume measurement.

6.  The recommended  temperature program  settings in the  method  should be
    converted  to  those presented  in the experimental  section  of  this re-
    port.   These conditions were established for analysis with tridecane as
    the solvent.

7.  Lower  limits  of detection  can be achieved by allowing the analyst to
    concentrate the final  extract  to as low as 10 pL.  It may be necessary
    to use the smaller final volume with other HRMS instruments to achieve
    the same levels of detection.

8.  The method should recommend several  techniques  to break  up emulsions
    resulting  from extraction of aqueous samples.  In  this evaluation the
    emulsion phase was put through a column packed with glass wool, which
    was  then  rinsed  with additional  metbylene  chloride.  Other  options
    might include stirring or centrifugation of the emulsion phase.

9.  The method should specify  the procedure to  deal with aqueous  samples
    containing high levels  of  suspended  solids.  In  this  study  it was

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     necessary to centrifuge  the  soil extract sample before proceeding with
     the extraction.

10.  It is highly recommended that the method be modified such that the ben-
     zene extract is  completely  exchanged to hexane prior to cleanup on the
     silica column since this is  apparently one of the most critical factors
     leading to successful sample analysis.

11.  It may be worthwhile to evaluate a cleanup procedure in which the char-
     coal column  precedes the alumina  column as a means  to  improve method
     recovery.

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                                  SECTION 4

                           EXPERIMENTAL PROCEDURES
SAMPLE DESCRIPTION

     Five  solid  samples  were  provided  by the  Environmental  Monitoring
Systems  Laboratory-Las Vegas  (EMSL-LV)  to  the  Midwest Research  Institute
(MRI)  for analysis  for  2,3,7,8-TCDD  and total  TCDD using the  analytical
method in Appendix A.  A description of the five solid samples  and the esti-
mated  2,3,7,8-TCDD  concentrations from previous analyses by an independent
laboratory are provided in Table 1.   Each sample was analyzed  in triplicate
as specified  in  the  protocol.  One of the triplicate samples  for each soil
sample was spiked with the seven TCDD isomers (1,3,6,8-; 1,3,7,9-; 1,2,3,7-;
1,2,3,8-;  1,2,3,4-;  1,2,7,8-; and  1,2,8,9-TCDD)  at  approximately  10 times
the estimated level of 2,3,7,8-TCDD specified in Table 1.

     Five aqueous samples were generated  for the evaluation of the analyt-
ical method at  the  ppq detection level.   Table 2 presents  a  description of
each water type and lists the fortification levels of 2,3,7,8-TCDD and seven
additional  TCDD isomers  (1,3,6,8-;  1,3,7,9; 1,2,3,7-;  1,2,3,8-;  1,2,3,4-;
1,2,7,8-; and 1,2,8,9-TCDD) in each sample.

     The  influent and  effluent  wastewater  samples  were  collected  from  a
sewage treatment facility in metropolitan Kansas City, Missouri.  The indus-
trial  wastewater  was  obtained from a holding pond  within  a hazardous waste
area that was known  to be highly contaminated  with PCBs and possibly other
chlorinated  aromatic compounds  (chlorobenzenes).   This aqueous  sample  was
very acidic (pH < 1) and was dark in color.

     The soil extract was prepared from 30 g of a soil sample,  Hyde Park 002
(H2),  and  1 gallon of distilled water.  The mixture  was stirred constantly
(at least 24 hrs) until just prior to subsampling of  1.0-L aliquots.

SAMPLE PREPARATION

     All  samples  listed  in  Tables  1  and  2  were extracted and analyzed in
triplicate according  to the protocol provided  in Appendix  A.   As indicated
in  Tables 1  and  2,  one  aliquot  of  each  sample  matrix was fortified with
additional  TCDD  isomers,  which  represent the  compounds  that  elute first
(1,3,6,8-TCDD),  last (1,2,8,9-TCDD),  and  within the  approximate retention
window of 2,3,7,8-TCDD (1,2,3,7-; 1,2,3,8-;  and 1,2,3,4-TCDD)  from the HRGC
columns used for sample analysis.

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        TABLE 1.  SOLID SAMPLES USED FOR HRGC/HRMS METHOD EVALUATION


Approximate
sample
EPA sample no. Matrix size3
B25-Piazza Road (B5) Soil 10 g
Hyde Park 001 (HI) Soil 10 g
B52-Shenandoah (Bl) Soil 1 g
Hyde Park 003 (H3) Soil 1 g
RRAI-5,7,8 (FA) Fly ash 10 g
Estimated
2,3,7,8-TCDD
concentration
(FPt)5
50
70
360
1,700
NRd

Spike level
(ppt) of
TCDD isomers
100
140
720
1,700
e

.Approximate sample size of each replicate sample.
 Estimated level of endogenous 2,3,7,8-TCDD reported to MRI by
CW. Beckert in letters dated April 19, 1985 and August 30, 1985.
 .Approximate fortification level of each of seven additional TCDD isomers.
^o estimate of 2,3,7,8-TCDD concentration was reported.
Additional TCDD isomers were not spiked into this matrix.
      TABLE 2.  AQUEOUS SAMPLES USED FOR HRGC/HRMS METHOD EVALUATION

Sample type
Distilled water (DW)
POTW influent (IWW)
POTW effluent (EWW)
Industrial wastewater (IND)
Hyde Park 002; soil extract

Approximate
sample
size
1.0 L
1.0 L
1.0 L
1.0 L
(H2W) 1.0 L
Fortification
level of
2,3,7,8-TCDD
(ppq)
250
500
1,000
500
c
Fortification
level of
TCDD isomers
(ppq)B
500
1,000
2,000
1,000
c

a
.Approximate sample size of each replicate sample.
 Approximate fortification level of each of seven additional TCDD isomers
 This aqueous sample was not fortified with TCDD isomers.

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     All  samples were  fortified with  500 pg 13C12-2,3,7,8-TCDD  in  1.5 ml
acetone.   The  solid  samples were  extracted  continuously  for  24 hr in  a
Soxhlet  apparatus  with benzene  and  the  1.0-L  aqueous samples  were  batch-
extracted using 2.0-L separately funnels and three 60-mL portions of methy1-
ene chloride.  The extractions of the influent wastewater (IWW) and effluent
wastewater  (EWW) and  the  soil extract  (H2W) were  complicated  by the forma-
tion  of emulsions.   In each case,  the  emulsion was removed by passing  the
methylene  chloride  and emulsion  layer  through a column of glass  wool pre-
rinsed  with methylene chloride.  The extract and  resulting aqueous  layers
were collected in a sample bottle and the glass wool plug was  rinsed with an
additional  10 ml methylene chloride.  Following the complete  extraction of
the aqueous  sample,  the  contents of the  bottle  were transferred to a clean
250-mL  separatory  funnel  and  the methylene chloride  was  removed  from  the
aqueous  phase that was transferred  with  the  emulsion.   All  extracts were
concentrated  with  Kuderna-Danish  evaporators  and  nitrogen evaporation to
approximately  1.0 ml.   Each  extract was  taken  through the entire cleanup
procedure  including  the  acidic silica,  acidic  alumina,  and  Carbopak C as
specified in  the protocol (Appendix A).  The HRGC/HRMS analysis of each  ex-
tract was completed as specified below.

REAGENTS

     All solvents for  extraction and cleanup were  obtained as "Burdick  and
Jackson  distilled-in-glass"  quality.  The tridecane (99 percent purity)  was
obtained from Aldrich (TS,  740-1).   The  chromatographic materials,  acidic
alumina  (100-200 nesh AG-4,  Biorad  Laboratories  132-1340), silica (70-230
mesh Kieselgel 60, EM Reagent, American Scientific Products  C5475-2), sodium
sulfate, potassium  carbonate, Celite 545®  (Fisher  Scientific  Company),  and
the silanized glass  wool  and Carbopak C  (80-100 mesh Supelco 1-1025) were
prepared for use as  specified  in Section 7 of  the protocol  (Appendix  A).

     Table  3  provides  the sources of standards used to prepare the calibra-
tion  solutions, sample fortification solutions, recovery  standard spiking
solution,  internal  standard  spiking solutions,  field  fortification solu-
tions, and TCDD isomer fortification solutions.

     Table  4 is a summary of  the concentration calibration  standards pre-
pared for the HRGC/HRMS method evaluation.  These standards  were prepared as
specified in  the protocol (Appendix A).  The standard HRCC6 was included in
the final  evaluation  of  the HRGC/HRMS method  as a means to demonstrate  the
lower limit of detection under optimum instrumental conditions.

HRGC/HRMS INSTRUMENTATION

     Sample  extracts  and  calibration standards were analyzed  using a Carlo
Erba Mega  Series gas  chromatograph (GC)  which was  coupled  to  a Kratos MS50
TC double-focusing mass spectrometer (MS).  The GC/MS  interface was simply a
direct  connection of  the  GC column to the ion source via a  heated interface
oven.   A Finnigan  2300 Incos data system was  used for data acquisition  and
processing.

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         TABLE 3.  TCDD ISOMERS USED FOR HRGC/HRMS METHOD EVALUATION
       Isomer
      Stock
  concentration
     Source
Standard code
2,3,7,8-TCDD


13C12-2,3,7,8-TCDD


1,2,3,4-TCDD


13C12-1,2,3,4-TCDD


1,3,6,8-/1,3,7,9-TCDD


1,2,3,7-/1,2,3,8-TCDD


1,2,7,8-TCDD


1,2,8,9-TCDD
Column performance
standard
7.87 ± 0.79 Mg/mL

  50 t 5 Mg/mL

    2.7 mg/mL

  50 ± 5 jJg/mL

    0.82 mg/mL

    0.5 mg/mL

    0.39 mg/mL

    1.46 mg/mL

   10 |Jg/mL
EPA QA Reference
Materials

Cambridge Isotope
Laboratories

Cambridge Isotope
Laboratories
Cambridge Isotope
Laboratories

Cambridge Isotope
Laboratories
Cambridge Isotope
Laboratories

Cambridge Isotope
Laboratories

Cambridge Isotope
Laboratories

Cambridge Isotope
Laboratories
20603

R00201 (Lot
AWN-1203-65)
ED-915C (Lot
6578)

R00212 (Lot
AWN-1203-93)
ED-913C (Lot
F2086)

ED-905C (Lot
7371)

ED-915C (Lot
7184)

ED-916C (Lot
MLB-682-26)

ED-908 (Lot
No. R00215)
 Mixture of TCDD isomers including 2,3,7,8-; 1,2,3,4-; 1,2,3,7-/1,2,3,8-;
 1,2,7,8-; and 1,4,7,8-TCDD.
    TABLE 4.  COMPOSITION OF CONCENTRATION CALIBRATION SOLUTIONS (pg/pL)

HRCC1
HRCC2
HRCC3
HRCC4
HRCC5
HRCC63
Recovery standard
13C12-1,2,3,4-TCDD
2.5
5.0
10.0
20.0
40.0
1.0
Analyte
2,3,7,8-TCDD
2.5
5.0
10.0
20.0
40.0
1.0
Internal standard
l3C12-2,3,7,8-TCDD
10.0
10.0
10.0
10.0
10.0
10.0

 This solution is not specified in the analytical method in Appendix A.
                                      10

-------
     The HRGC/HRMS operating conditions used in the final phase of this work
are summarized  in  Table 5.   The GC operating  conditions  recommended  in the
protocol were  not used  for these analyses  for three  reasons.   First,  the
TCDDs hr.ve rather long retention times, and the solvent (tridecane) boils at
235°C.   Thus  no benefit  could be realized with a  low initial temperature.
Second,  past  experience at  MR I has indicated  that 200°C  is  an acceptable
starting temperature  for  these types  of analyses when  tridecane  is used as
a  solvent.   Finally,  since  the CP-Sil 88 and SP-2330 phases  are both very
polar and  thinly coated,  it has been recommended that they not be subjected
to  rapid  heating or  cryogenic  cooling  to prevent  thermal  shock to  the
column.3

     The MS  was tuned daily to yield  a resolution of at  least  10,000 (10
percent valley) and optimal response at m/z 254.986.  This step was followed
by calibration of an accelerating voltage scan beginning at m/z 254 (typical
calibration range  was 255  to  605 amu).  Other voltage scans  from the same
data  file  were  then  used  to establish and document both  the  resolution at
m/z 316.983 and the mass measurement accuracy at m/z 330.979.

MASS MEASUREMENT ACCURACY

     For this work, mass  measurement  accuracy was  measured  relative  to PFK
m/z 254.986, as  required  by the protocol, by  applying the mass correction,
Am, to  the  entire spectrum, which yields an error of 0 ppm at m/z 254.986.
In this  way,  it was possible to meet routinely the 5 ppm accuracy criterion
at m/z  330.979.  However,  if  a  lock  mass  other than 254.986  is used,  the
mass  measurement accuracy should  be  measured  relative  to that  lock  mass,
since it is that peak  which is used  to maintain magnet  alignment and will
ultimately control  the  mass measurements during the selected ion monitoring
(SIM) experiments.

Mass Resolution

     Mass resolution at m/z 316.983 was documented by an output of the Incos
PROF  program.    However,  the  computer-generated value  for resolution  was
found to be significantly  higher than the value  measured manually.   Thus,
the manually determined  resolution,  which was nearly identical to the value
measured by using the peak matching unit, is reported.  Closer inspection of
the PROF source  code  revealed  that resolution is computed via a statistical
method,  not  as  m/Am  at  5  percent height.  Incos users  should therefore be
aware of this discrepancy,  because  the computer-generated  value can  be as
much as 20 percent over the proper value.

     Following calibration, the SIM experiment descriptor was updated to re-
flect the new calibration.  Six masses  (see Table 5) were monitored by scan-
ning ^ m/10,000 amu over each mass.  The total cycle time was kept to 1 sec.
The m/z  280.983  ion from PFK was used as a lock mass because it is the most
abundant PFK  ion within the range  of  m/z 255  to 334  and therefore permits
the use  of  low partial pressures of PFK, which minimizes PFK interferences
at the analytical masses.
                                      11

-------
                  TABLE 5.  HRGC/HRMS OPERATING CONDITIONS
Mass spectrometer
          Accelerating voltage:
          Trap current:
          Electron energy:
          Electron multiplier voltage:
          Source temperature:
          Resolution:

          Ions monitored

             258.930
             319.897
             321.894
             331.937
             333.934
             280.9825 (lock mass)
          Overall SIM cycle time

Gas chromatograph

          Column coating:
          Film thickness:
          Column dimensions:

          Helium linear velocity:
          Helium head pressure:

          Injection type:
          Split flow:
          Purge flow:
          Injector temperature:
          Interface temperature:
          Injection size:
          Initial temperature:
          Initial time:
          Temperature program:
= 1 sec
           8,000 V
             500 MA
              70 eV
           2,000 V
             280°C
          10,000 (10% valley definition)

           Nominal dwell times (sec)

                     0.15
                     0.15
                     0.15
                     0.15
                     0.15
                     0.10
           CP-Sil 88
           0.2 pm
           50 m x 0.22 mm ID

           * 25 cm/sec
           1.75 kg/cm2 (25 psi)

           Splitless, 45 sec
           30 ml/min
           6 ml/min
           270°C
           240°C
           2 HL
           200°C
           1 min
           200°C to 240°C at 4°C/min
                                      12

-------
CHROMATOGRAPHIC RESOLUTION
                                         .                      «
     Chromatographic resolution values were measured for the SIM plot of m/z
320.  However,  it  may be advantageous to measure chromatographic resolution
from a plot  of the sum of m'z  259,  320, and 322.  The sum trace has better
signal-to-noise  ratio (S/N)  and  peak definition than the  SIM plots,  which
permits a more accurate measurement of resolution.

Selection of the HRGC Column

     Three different HRGC columns were evaluated in the course of this proj-
ect:  SP-2330  (60  m x 0.24 mm); DBS (60 m x 0.22 mm); and CP-Sil 88 (50 m x
0.22 mm).  By  evaluating the mass spectra  of  the bleed from each column at
240 to 250°C,  it became apparent that the column background may be the lim-
iting factor in achieving the desired detection limit for this method.  The
DBS column provided the least amount of background at 250°C, and the SP-2330
had the worst.   This coincides with the fact that quantitation at the detec-
tion limit (i.e.,  2.5 pg/|JL)  with the SP-2330 column was difficult at best.
The CP-Sil  88   column  appeared  to offer less bleed than  the  SP-2330 column
and indeed does permit more  accurate quantitation due to reduced background
contribution.

     The chromatographic  performance  afforded  by these columns is a further
issue, since the column best suited for low detection limits,  DB-S, cannot
meet the 25 percent valley chromatographic resolution criteria in all cases.
Both the SP-2330 and CP-Sil  88 columns can easily resolve the 2,3,7,8-TCDD.
However, based  on  the bleed  considerations discussed above, the 50-m CP-Sil
88  column  is  recommended for the  best combination  of  low bleed  and good
isomer separation.

     It may  also be  advisable  that other  HRGC  columns  (including SP-2340,
Silar IOC,  and SP-2331) that have been used for 2,3,7,8-TCDD analysis at the
1-ppb soil level be evaluated for background contribution and their applica-
tion for HRMS analysis at ppt and ppq concentrations.

INJECTION TECHNIQUE

     The HRGC  column performance  can degrade very quickly  if proper injec-
tion  techniques are  not used.   Specifically,  the  SP-2330  and  CP-Sil  88
phases are very sensitive to ©2  and  will  decompose  rapidly at 200°C if any
trace of 02 is present.  Therefore, the common practice of using 1 pL of air
to  flush  the syringe  and effect reproducible injections is  to  be avoided,
since even that small amount of  air per injection can cause column perfor-
mance to degrade in less than one week of continued use.

     The following  injection  technique  is recommended.  First rinse the sy-
ringe copiously with isooctane (or other volatile solvent, such as toluene).
Dry the syringe by drawing air through  it.  Pull up  and expel several vol-
umes of tridecane until all bubbles are gone, and leave 1 pL of tridecane in
the barrel.  Finally,  pull  up 2 pL of the sample solution and inject.  This
technique has worked very well  and yields injection reproducibility compar-
able to that of the air purge method, without introducing air onto the ana-
lytical GC column.

                                      13

-------
                                  SECTION 5

                           RESULTS AND DISCUSSION

     The primary purpose  of any method validation process is to assure that
the  method  under consideration  is  adequate to meet  testing and monitoring
requirements.1  The  single-laboratory  evaluation  of the analytical protocol
presented in  this  report  has been preceded  by several  evaluation  and im-
provement steps.  These have included the preparation of a written protocol,
technical review of  the protocol for  completeness,  technical  accuracy, and
clarity;  preliminary  testing  to  evaluate  performance  of the  analytical
method;  and  revision and  refinement  of the  written protocol  based on the
results of the preliminary testing.

     Prior to the assessment of the refined protocol presented in Appendix A,
the  proposed  analytical method  had  been evaluated  for performance  through
the  analysis  of several duplicate samples.   The results  of the preliminary
evaluation indicated that problems existed in the design and approach to the
extract  cleanup steps, which  greatly  affected the  method  detection limit,
accuracy, and precision.

     This section  presents a  summary  of the  studies that  have  led to the
refinement of the  analytical  protocol as provided  in Appendix A and  also
summarizes the single-laboratory evaluation of this protocol.

APPROACH TO CLEANUP COLUMN EVALUATION

     The initial method evaluation completed under the  first  task resulted
in very low recoveries of the internal standard, 13Cl2-2,3,7,8-TCDD,  and the
accuracy and  precision of  duplicate  sample analyses were  poor.   After re-
viewing  the data,  it was apparent that the problems were the result of poor
chromatographic separation in the cleanup columns.   The initial protocol in-
volved reducing sample extract volumes to 1.0 mL in benzene, elution through
the  acidic  silica  column  with hexane, and  collection of  the  total eluent
which was then  added to the acidic alumina  column.   The alumina column was
further  eluted  with hexane/20-percent  methylene  chloride.   The  eluate was
concentrated  and cleaned further using a Carbopak C/Celite  column,  and the
TCDDs were eluted with 2 mL toluene.

     Column cleanup  techniques  were  revised and further evaluated following
the  procedures  depicted in  Figures  1  and  2.   The  column  evaluations  were
completed with triplicate measurements at three spike levels (0.10, 1.0, and
10 ng) equivalent  to 10,  100, and 1,000 ppt of TCDD in solids with several
TCDD  isomers  (2,3,7,8-;  1,3,6,8-;  1,3,7,9-; 1,2,3,4-;  1,4,7,8-;  1,2,3,7-;
1,2,3,8-; and 1,2,8,9-TCDD).

                                      14

-------
           OPTION A
        1 ml Benzene Exfrocf
              I
          H2SO4 - SJO2
          4.0g
             Si 02
              l.Og
          Acidic AI2O3
          6.0g
                30ml 20% CH2Cl2/H«xon«
       Concentrate fo
              I
        Corfaopok C/C*lit«
                6ml Toluene
            HRGC/HRMS
   OPTION B
1 ml Benzene extract
       I
  H2SO4 - SIO2
  4.09
      SiO2
      l.Og
  Acidic AI2O3
  6.0g
                                             Carfaopok C/C«m«
        6 mi Toluene
    HRGC/HRMS
                                                                      •xan«
Figure  1.    Column  cleanup  procedures  specified   in  the  protocol

                                    15

-------
          OPTION C
      1 ml Benzene Extract
H2SO4 - SiO2
4.0g
/.Og
4
>
Concentrate fo
0.5mL
             I
         Acidic AI2C>3
         6.0g
               30 mC 20% CH2Cl2/H«*ane
      Concentrate fo
             I
       Corbopak C/Cei!te
               6mL Toluene
          HRGC/HRMS
    OPTION 0
I ml Senzene Extract
                                              H2S04 - Si02
                                              4.09
                                                 Si02
                                                 l.09
                                           Concentrate to 0.5mL
       I
   Acidic AI2O3
   o.Og
         30mL 20% CH2Cl2/Hexane
 Carbopok C/CeHte
         6mi Toluene
                                               HRGC/HRMS
Figure 2.   Column cleanup  procedures  proposed by the EMSL-LV.

                                    16

-------
     The TCDD  isomers  were added to 1-mL portions of benzene and were taken
through the  four sample cleanup sequences depicted in Figures 1 and 2.   One
of  the  replicates  for  each procedure was also spiked with 100 ng of Aroclor
1260.

     The results of the sample analyses are provided in Tables 6 through 9.
As  noted  in Tables  6  and  7,  recoveries of  the TCDD  isomers  were  low and
quite variable for the  early eluting isomers  1,3,6,8-  and  1,3,7,9-TCDD as
compared to  1,2,8,9-TCDD.   Recovery of 1,2,8,9-TCDD was still low and vari-
able (approximately 60 percent recovery with an RSD of * 20 percent).  These
results were generated using the procedures specified in the original proto-
col  (see Figure  1).  The results of the  analyses  following  the cleanup op-
tions A and  B  demonstrate that accurate quantitation of all TCDD isomers is
not possible using only the 13Cl2-2,3,7,8-TCDD surrogate standard.  The low
recoveries measured  for options A and B are obviously a result of the pres-
ence of benzene  in the eluent from  the  acid-modified silica column that is
taken directly through the acidic alumina column.

     In contrast,  options  C and D (Tables 8 and 9) demonstrate quantitative
recovery of  the  TCDD isomers.  Some background contamination has been noted
from the  acidic alumina  for the  1,3,6,8-  and  1,3,7,9-TCDD  isomers.   This
material had previously been prepared by Soxhlet  extraction with methylene
chloride and activation at 190°C prior to use.  As noted in Tables 8 and 9,
the  average  recovery of  the other spiked TCDD  isomers  was  greater  than 84
percent.

     When the recoveries of the different isomers and the 13Ci2-2,3,7,8-TCDD
are compared,  the  average relative percent difference ranges from 1 percent
for 2,3,7,8-TCDD  (Table 3)  to 24 percent for 1,2,3,4-TCDD (Table 4).  These
results demonstrate that either of  these cleanup  procedures (options C and
D)  will provide  good recovery and reliable quantitation of 2,3,7,8-TCDD and
very good  estimates of the concentrations of the other TCDD isomers present
in  the  samples.   No interferences were observed in  the samples spiked with
100 ng Aroclor 1260.   The lack of PCB interferences was especially noted in
the extracts of samples spiked at 0.10 ng/TCDD isomer.

     In addition to the evaluations of the cleanup procedures presented above,
the acid-modified silica gel/acidic alumina columns and the Carbopak C/Celite
column were  evaluated  separately.  Evaluation of  the silica/alumina at the
0.10-ng spike  level as shown in Figure 2 resulted in an average recovery of
120 percent  for  1,2,3,4-,  1,2,3,7-,  1,2,3,8-, and 1,4,7,8-TCDD; 114 percent
for  2,3,7,8-TCDD;  118  percent for  13Cl2-2,3,7,8-TCDD;  and  118 percent for
1,2,8,9-TCDD.   The results  for the recovery of  1,3,6,8-  and 1,3,7,9-TCDD
indicated that some contamination originated from the acidic alumina.

     Replicate analyses of the Carbopak C/Celite column at the 0.10-ng spike
level resulted in average recoveries of 97 percent for 1,3,6,8-TCDD; 88 per-
cent  for  1,3,7,9-TCDD;  81 percent  for  1,2,3,4-,  1,2,3,7-,  1,2,3,8-,  and
1,4,7,8-TCDD; 75 percent for 2,3,7,8-TCDD; 96 percent for 13C12-2,3,7,8-TCDD;
and  90 percent for 1,2,8,9-TCDD.   Elution  of  the Carbopak C/Celite column
with additional  toluene beyond 6 ml did not improve recoveries even for the
samples spiked at  10 ng/TCDD isomer.

                                      17

-------
                                             TABU 6.  KECOVKKV  (1)  OF SEVERAL TCIM)  ISWIKRS H!UM CI.EANIII* 01TION  A
00
_..... _ .1 1 —




l.l»*M~.t.l..f
1
HJSO« - siQ}
4-0.
$10}
i
AcMUAIjOj
iJO.t M«
CMMWMM* «• IflO/it
*

U.I IclMK
HRGC/IIRMS



Spike
level 1,3,6,8


1 ng 4.6
1 it a 12
1....* •» •

10 ug 12
10 ng 6.2
""" 10 ng' 6.3

He a u 7.4
IO »L
2,3,7/
2,3,8


14
ll
• 7

31
20
20

23





TCDI) iioaei
2,3,7,8


19
11
94

44
31
29

31





r
l3C12-2.3l7.ft


27
4B
11

38
36
34

36






1,2,8,9


38
64
An

76
61
59

56




                          Sanple  was also spiked with Kill ng »l Arorlor  IJ60.

-------
                    TABI.K 7.  RECOVERY  (1) OF SKVKRAI. TCDI1  ISOHKRS FROM CLEANUP OPTION B

Recovery (%) of TCOI) I»CNK

1 .1 I>M«M tmUta
J
MjSO< - JIOj
4.0,
IIOj
, 1
A*l*c AljOj
•Of
130.1 »«
Cntijil C/C.III,
Spike
level 1,3,6,8
1 ng 7.6
1 ng 2.4
1 ng* 6.4
10 ng 0.9
10 ng 1.8
CHjCiykum
10 nga II
J».tl.h.M
Mean 5.0
HRGC/HRMS

1 USD 79
1,3.7.9
15
21
25
7.6
II
17
16
40
I.2.3.4/
1,4.7.8
35
31
34
14
36
44
32
31
I.2.3.7/
1.2,3,8
14
17
22
21
25
31
22
28
2,3.7,8
37
34
40
50
47
59
45
21
r
'3Clj-2,3,7,8

38
40
41
44
48
52
44
12

1,2,8,9

48
52
57
70
59
78
61
19

JSan|)le was also spiked witli  100 ng ol  Arurlor 1260.

-------
                                           TAHIJi 8.  kecOVKKY  (1) UK SKVEKAI. TCOO ISONEMS FKOH C.l£MtUP OPTION C
ro
O
, ..._.,_. . ..... , . , ___________ 	 _ 	 . 	 __
Recovery (1) of TCI1I) inomfr
Spike
level 1.3.6.8"
1 «4 •••••<• I rtrari


M,I04 . JIO,
SIOj
1

J
AtMcAJjOj
Itt»t30%
C~«*«.I.MOjU
J
c.wu eye.**
1 4«t I.I-.
HRGC/HRMS
0
0
0

1
1
1
10
citjciykiuw
10
10
. 10 ug
.10 ng
. 10 ugd
.0 ng
.0 ng
.0 ngd
.0 ng
.0 ng
.0 ngj
Mean
m
, .

300
310
340
158
168
171
157
130
210
39
1
1.3.7,9" 1
390
420
440
165
162
183
155
140
130
240
54
.2.3,47
,«./,8
l'"u"
97"
140
98
136
118
130
102
115
14
1.2,3,77

c
c
c
116
96
120
116
130
99
112
12


107
102
88
94
80
96
110
130
100
101
14
'C|2*2 378

89
89
92
95
75
90
88
106
85
90
9.1
1289

81
84
91
89
80
94
107
122
110
95
15
                       *Tlie  1,3,6,8-  and 1,3,7,9-TCOD iuoaers were also noted in reagent blanks fro» the acidic  aluaina
                         coluwi.   No «ucIt interferences we-re noted fron tlu> dcidificd Milieu gel or the Carbopak  C/Celite
                       . coluwi.
                         Resolution of 1,2,3,4-,  1,2.J,7-/1.2, I.8-, and 1,4,7,8-TCIM} was not achieved.  This value
                         represents recovery of the four isoaers.
                       ^Recovery reported with I ,^,3,4-/l,4,7,fl-TCIin.
                         Sa>ple was also  spiked with 100 ng of Aroclor  1260.

-------
                    TABLE 9.  RKCOVtKY  (1) OF SEVERAL TCIID ISOHERS FKOM Ol.tAHUI' OITION D

Recovery (D ol
Spike
level 1

1 «l (••»« biracl
I
H,5O< - JIO,
4.0.
JIOj
1.0,
1

I
AcMkAljOj
«.o.
1 M»l 10%
C.WH.C/IC.MI.
1 *ml T«h»
HRGC/HRMS
0
0
0
1
1
10
CHaCI»ftln»m If)
10
M
H
t

.10 ng
.10 ng
. 10 ng*1
.0 ng
• 0 ng
.0 ,,gd
.0 ng
.0 ng
.Ongd
can
RSI)
.3,6,8'
147
290
260
90
180
135
81
114
126
158
46
1 1 1 9s 14

197
200
360
93
185
158
107
72
138
168
51
,3.4/
7 8

in"
•21"
nob
106
95
85
50
105
89
97
21
TCDI) iiioaer
1,2, 3,77

c
c
c
103
121
79
104
108
95
101
14

72
71
84
85
109
47
101
107
91
85
23

C12-2.3.7,8
84
90
97
86
90
60
78
82
92
84
13

1.2.B.9
70
71
e
53
108
63
84
118
112
85
29

 The 1,3,6,8- ami  1,3,7,9-T4'l)0 f suiters were also noted  in  reagent blanks fro* Hie acidic (liraina
 coluMi.  No such  in! i:r liMern es were nutnl 11 un I lie  uciililied silica gel or Hie Carliopdli C/Celitc
. colimn.
 Resolution ol  I.2.J.4-,  1,2,:i,7-/l .2,3,8- , and I ,4, 7,B-TCI)I> was not achieved.  This value
 represents recovery  ol  I lie lour isowrH.
^Recovery reported with  I,2,3,4-/l,4,7,8-TCW).
 Sunple was also spiked  with 100 ng of Aroclor 1260.
 MHIJC/IIKHS analysis was  interrupted pi ior to Hie olutiun of  lliis isnner.

-------
     Three additional  experiments  were completed to evaluate the efficiency
of reverse elution of the carbon  column.  The  Carbopak C/Celite was placed
in a 5-mL disposable pipette packed at both ends with glass wool plugs.  The
column  was  eluted  in one direction  for the hexane,  cyclohexane/methylene
chloride, and  the methylene  chloride/methanol/benzene  mixture.   The column
was then  turned  over and eluted with  6 ml toluene.   Triplicate analyses at
the 0.10  ng/TCDD isomer  spike  level  demonstrated average  recoveries  of 98
percent  for  1,3,6,8-TCDD;  91 percent  for   1,3,7,9-TCDD;  104 percent  for
1,2,3,4-,  1,2,3,7-,   1,2,3,8-,  and 1,4,7,8-TCDD;  116 percent  for  2,3,7,8-
TCDD;  102 percent for 13C12-2,3,7,8-TCDD; and  93 percent  for 1,2,8,9-TCDD.

FINAL METHOD EVALUATION

     Based on  the results of the  column  evaluation study,  the  analytical
method was revised to specify the cleanup procedure presented as Option D in
Figure 2.  The final  protocol,  as presented in Appendix A, was  then evalu-
ated as described below.

     The data presented in Tables 10 through 17 are summaries of the initial
column calibration, HRGC  and HRMS resolution checks, and the results of the
sample analysis.

Calibration

     Table  10  summarizes  the RRF data  for  the concentration  calibration
standards from the initial calibration and the routine monitoring of the RRF
values over the  time  required to complete the  sample  analyses.   The RRF(I)
as specified  in  the  protocol  is a measure of  the response of 2,3,7,8-TCDD
versus the internal standard, 13Ci2-2,3,7,8-TCDD.  The value for RRF(I) var-
ied ±9.4 percent over the five concentration levels  of 2,3,7,8-TCDD ranging
from 2.5 pg/pL to 40  pg/pL.   The RRF(II) is used to calculate the absolute
recovery  of  the  internal standard as  compared to  the recovery  standard
13C12-1,2,3,4-TCDD.   The  average RRF(II) was  determined to  vary by ± 19.3
percent  over  the  calibration curve.   The variability of  the  RRF(I)  and
RRF(II)  were   determined   to  be  less  than ± 10 percent and  ± 18 percent,
respectively,  over all data points required to complete the sample analysis.

     In addition  to  the  analysis of calibration standards  specified  in the
protocol, solution HRCC6  was analyzed in triplicate to  determine the  lower
limit of  sensitivity  (1 pg/pL).   Although the calculated RRF(I) and RRF(II)
values and the S/N are within the specified criteria, the ion ratio for the
native compound  and  recovery standard indicate that these measurements fall
outside the acceptable calibration window.
                                      22

-------
ro
to
TARU 10
Calibration
atandard
imcci
imcci
IIRCCI


mice*
IIRCC2
IIRCC2
IIRCC2


HRCC3
IIRCC3
HRCC3



HRCC4
HRCC4
NRCC4


IIRCC5
HRCC5
IIRCC5




IIRCC6
HRCC6
IIRCC6
HRCCI
IIRCCI
IIRCC2
IIRCC2
IIRCC2
imcc2
HRCC2
IIRCC2
IIRCC2
IIRCC2
IIRCC2

Date
09/12/85
09/12/85
09/12/85


09/12/85
09/12/85
09/12/85
09/12/85


09/13/85
09/13/85
09/13/85



09/13/85
09/13/85
09/13/85


09/13/85
09/13/85
09/13/85




09/16/85
09/16/85
09/16/85
09/16/85
09/16/85
09/20/85
09/23/85
09/23/85
09/24/85
09/25/85
09/26/85
09/27/85
09/30/85
10/03/85

Tlae
09:05
09:44
12:31


13:00
13:27
13:53
15:39


10:31
10:57
11:23



13:02
13:29
13:56


14:22
14:49
15:15




10:43
11:19
13:44
12:42
14:40
10:46
08:47
10:46
10:47
08 : .19
08:56
09:33
09:19
08:53


•/» 320/322 •/*
0.
0.
0.


0.
0.
0.
0.


0.
0.
0.



0.
0.
0.


0.
0.
0.




,
1.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
82
84
84


73
80
92
78


78
78
73



77
73
77


78
75
76




32
18
86
87
81
86
82
88
89
76
77
78
83
68


332/334
0.
0.
0.


0.
0.
0.
0.


0.
0.
0.



0.
0.
0.


0.
0.
0.




0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
77
80
73


77
73
66
77


7»
80
83



80
76
78


80
82
78




71
8.1
80
8J
79
73
80
80
69
78
80
84
80
75
. IMITIAL CALIBRATION SUHHART



(IS) aj/z 332/334 (RS) S/H 259 S/N 322 S/N 334(IS) RJ)F(I)
0.
0.
0.


0.
0.
0.
0.


0.
0.
0.



0.
0.
0.


0.
0.
0.




0.
1.
1.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
80 41:1 > 65:
73 48:1 > 65:
87 24:1 > 65:


83 36:1 > 65:
81 58:1 > 65:
69 78:1 > 65:
73 76:1 > 65:


79 97:1 > 65:
78 111:1 > 65:
78 110:1 > 65:



76 96:1 > 65:
78 > 144:1 > 65:
76 > 144:1 > 65:


78 > 144:1 > 65:
83 > 144:1 > 65:
79 > 144.1 > 65:




84 10: 21:
14 9.6: 25:
01 18: 30:
75 36: 63:
85 48: > 6.1:
80 > 75: > 63:
70 > 75: > 63:
83 42: > 63:
69 26: > 63:
89 58: > 63:
72 73: > 63:
81 49: > 63:
85 7.1: > 63:
81 29: > 63:
1 > 65:1 0.783
1 > 65:1 0.794
1 > 65:1 0.750
Nea> 0.776
t RSD 2.9t
1 > 65:1 0.829
> 65:1 0.853
1 > 65:1 0.799
1 > 65:1 0.861
Mean 0.848
X RSD 2.01
1 > 65:1 0.974
1 > 65:1 0.965
1 > 65:1 0.972
Head 0.970
1 RSD 0.5t
t
1 > 65:1 0.945
1 > 65:1 0.935
1 > 65:1 0.967
Mean 0.949
I RSO 1 . 7X
1 > 65:1 0.964
1 > 65:1 1.01
1 > 65:1 0.989
Mean 0.987
I RSD 2.2X
Overall llran (RRF) 0.906
X RSD 9.4X
> 65: 0.917
> 65: 0.878
> 65: 0.935
> 65: 0.876
> 65: 0.850
> 63: 0.835
> 63: 0.832
> 63: 0.941
> 63: 1.01
> 63: 0.949
> 63: 0.941
> 63: 1.04
> 63: 0.854
> 63: 0.955


RRF(II)
2
2
2
2
2
1
1
2
2
1
U
,
1
I
I
1

1
1
1
1
1
1
I
1
1
3
1
19
1
1
1
1
2
I
2
1
1
1
1
1
1
	
.18
.27
.28
.24
• 4X
.76
.93.
.03*
.27
.99
• OX
.53
.57
.58
.56
• 9X

.49
.52
.49
.50
• IX
.52
.42
.43
.46
• 9X
.75
• 3X
.44
.09
.58
.86
.11
.98h
.65
.41
.68
.96
.57
.68
.67
.53
          .Nnl  Inrliiilrd in «ran  RRK rnaniitat Ion.

           Nnl  witliin allowable  limit* lor  roiil iiir ral dual Ion.

-------
                    TABLE 11.  HRCC AND MASS RESOLUTION CHECK SUMMARY
Date
9/12/85
9/12/85
9/12/85
9/12/85
9/13/85
9/13/85
9/13/85
9/13/85
9/16/85
9/16/85
9/16/85
9/16/85
9/17/85
9/17/85
9/18/85
9/18/85
9/19/85
9/19/85
9/20/85
9/20/85
9/20/85
9/20/85
9/23/85
9/23/85
9/23/85
9/23/85
9/24/85
9/24/85
9/24/85
9/24/85
last.
ID
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
NSSO
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
NSSO
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
MSSO
Sol.
ID
.
PC
PC
-
-
PC
PC
-
-
PC
PC
-
PC
-
-
PC
-
PC
-
PC
PC
-
. •
PC
PC
-
PC
PC
"
Tiae
07:51
08:32
16:05
16:47
08:26
08:45
15:48
16:23
09:25
10:45
15:16
15:57
10:26
10:14
07:58
08:18
11:14
12:56
08:00
08:16
15:44
16:14
07:55
08:15
16:01
16:45
09:51
10:15
16:01
16:33
TCDD isomer
File resolution
name (% valley)
MID254I12X1
83671 12XQ1
83671 12XQ9
HID2S4I12X2
MID2S4I13X1
83671 13XQ1
83671 13XQ12
HID2S4I13X2
MID2S4I16X1
8367I16XQ1
83671 16XQ8
MID254I16X2
8367I17XQ1
MID2S4I17X1
MID254I18X1
8367I18XQ1
MID254I19X1
6367I19XQ1
MID2S4I20X1
8367I20XQ1
8367I20XQ5
MID254I20X3
MID254I23X1
8367I23XQ1
8367I23XQ6
Manual check*
MID2S4I24X1
8367I24XQ1
8367I24XQ3
MID2S4I24X2
-
5.9
2.9
-
-
6.9
11.4
-
-
11.9
23.0
-
13.3
-
-
20
-
3.5
-
6.7
4.1
-
-
8.8
12.5
-
12.2
13.1

Mass
resolution
at 10X valley
10,774
-
-
10,450
10,230
-
-
10,384
10.294
-
-
10,388
-
10,824
11,019
-
11,679
-
12,068
-
-
10,777
10,096
-
12,500
10,374
-
-
10,567
Mass
Measurement
error
5 ppn
-
-
-
0 ppm
-
-
-
4 ppn
-
-
-
-
2 ppm
1 ppm
-
4 ppn
-
3 ppm
-
-
-
1 ppn
-
—
3 ppa
-
-

*A aianual resolution cheek was performed due to data syitesi failure.
                                       (continued)
                                            24

-------
                                TABLE 11.   (continued)
Date
9/25/85
9/25/85
9/25/85
9/25/85
9/26/85
9/26/85
9/26/85
9/26/85
9/27/85
9/27/85
9/27/85
9/27/85
9/30/65
9/30/85
9/30/85
9/30/85
10/3/85
10/3/85
10/3/85
10/3/85
lost.
ID
HS50
HS50
MS50
HS50
MS50
HS50
MS50
MS50
HS50
MS50
HS50
HS50
HS50
MS50
HS50
MS50
asso
HSSO
.1S50
MS50
Sol.
ID
.
PC
PC
-
-
PC
PC
-
-
PC
PC
-
-
PC
PC
-
-
PC
PC
'
Tine
07:50
08:05
16:13
16:45
08:07
08:21
15:49
16:21
08:19
09:02
16:01
16:29
08:15
08:33
15:10
15:41
07:59
08:20
15:56
16:29
File
nine
HID254I25X1
8367I25XQ1
8367I25XQ3
MID254I25X2
MID254I26X1
8367I26XQ1
8367I26XQ3
MID2S4I26X2
MID254I27X1
8367I27XQ1
8367I27XQ3
HID254I27X2
MID254I30X1
8367I30XQ1
8367I30XQ3
HID254I30X2
MID254J03X1
8367J03XQ1
8367J03XQ3
HID254J03X2
TCDD if oner
resolution
(* vallev)
.
11.1
6.5
-
-
8.3
13.2
-
-
11.9
11.8
-
-
< 25
< 25
-
-
17
12
"
Mass
resolution
at 10% valley
11.165
-
-
11,419
10,989
-
-
10,499
11,564
-
-
10,639
11,149
-
-
11,321
10.567
-
-
10,442
Mais
Measurement
error
0 ppm
-
-
-
3 ppm
-
-
-
1 ppm
-
-
-
5 pp«
-
-
•»
0 ppm
-
-
'
A manual resolution check was performed due to data system failure.
                                           25

-------
                                                                                      TABLE  12.  TCDD DATA REPORT FORM
Aliquot
Sample Air-dry wt. (g) TCDD
Mo. or Vol. (L) Isomer
8367-83- 1576X-DVD 1.0 L
2,3,7
,8-
1,3,6,8-

8367-82-1576X-DV 1.0 L
8367-89- 1576X-EV.VD 1.0 L



8367-88-1576X-EWW 1.0 L

8367-90-1576X-EWN 1.0 L







8367-85-1576X-IND 1.0 L
8367-92- 1576X-IWWD 1.0 L
-


8367-87-1576X-INDN 1.0 L

8367-84- 1576X-DWH 1.0 L

-

1,3,7
2,3,7
2,3,7
1.3,6
1,3,7
c
2,3,7
c
2,3,7
1,3,6
1,3,7
1,2,3
1,2,3
1,2,3
1,2,7
1,2,8
2.3,7
2,3,7
1,3,6
1,3,7
1,2,7
2,3,7
1,2,7
2,3,7
1,3,6
1,3,7
1,2,3
,9-
,8-
,8-
,8-
,9-
TCDD (ppt
Retention time or ppq)
TCDD
21:35
16:43
17:54
21:35
21:40
16:48
17:58
24:10
,8-
21:29
23:57
,8-
,8-
,9-
,7/
,8-
,4-
,8-
,9-
,8-
,8-
,8-
,9-
,8-
,8-
,8-
,8-
,8-
,9-
,11
21:
59
17:02
18:14
22:

22:
24:
30:
21:
22:
17:
18:
24:
22:
24:
22:
17:
18:
22:
18

31
30
01
55
01
06
16
33
00
31
01
06
18
21
1JC,2-2,3,7,8 Meas. DL
21:34 228
117
86.5 -
21:33 196
21:37 2,277
134
282
137
21:27 1,090
75.9 -
21:56 1,010
502
766
1,860

1,840
3,430
1,330
21:55 1,290
21:59 508
191
208
55.2 -
21:59 1,520
586
22:00 234
512
395
403
HRGC/HRMS Analysis
Instr. Relative loo Abundance Ratios
ID Date Time 320/322 332/334(15)
MS50 09/20/85 11:27 0.
78 0.63
0.69
0.80
MS50 09/20/85 13:02 0.58 0.69
MS50 09/20/85 14:41 0.
1.
0.
0.
MS50 09/20/85 15:11 0.
0.
MS50 09/23/85 11:17 0.
0.
0.
0.

0.
0.
0.
MS50 09/23/85 12:51 0.
MS50 09/23/85 13:29 0.
0.
0.
0.
MS50 09/23/85 14:00 0.
0.
MS50 09/23/85 14:31 0.
0.
0.
0.
79 0.72
02
84
90
77 0.71
89
74 0.72
85
83
81

71
78
80
81 0.90
74 0.80
85
75
89
87 0.78
71
75 0.71
80
74
72
332/334(RS) X Rec. m/r 259
0.68 40 73:
64:
41:
0.72 14 21:
0.71 96 > 146:
10:
28:
6:
0.72 61 146:
7:
0.82 91 27:
22:
30:
54:

45:
78:
22:
0.81 23 37:
0.71 75 49:
25:
27:
8:
0.86 20 49:
21:
0.74 82 23:
73:
53:
42:
1
1
1
1
1
1
1
1
1
1
1
1
1
1

1
1
1
1
1
1
1
1
1
1
1
1
1
1
S/N
m/z 322 m/2 334(IS) Comments
> 63:
50:
30:
62:
> 63:
> 5:
> 11:
63:
> 63:
31:
34:
22:
33:
> 63:

60:
> 63:
18:
63:
63:
29:
29:
16:
63:
31:
.
63:
50:
45:
1 > 49:1 Ratio unacceptable. Rerun.
1
1
1 > 20:1 Ratio unacceptable; low recovery. Rerun.
1 > 63:1 Sample spiked at twice requested level.
1 Rerun.
1
1
1 > 63:1
1
1 > 63:1
1
1
1

1
1
1
1 30:1 Low recovery.
1 > 63:1
1
1
1
1 23:1 Low recovery.
1
> 63:1
1
1
1
1,2,3,8-
1,2,3,4-
1,2,7.8-
1,2,8,9-'
22:
24:
30:
35
32
02
616
840
328
0.
0.
0.
72
80
79
61:
70:
23:
1
1
1
55:
63:
20:
1
1
1
^Aqueous sample data reported as ppq and_soil sample data'presented as ppt.
 Criteria for positive identification require that the ion ratios fall between 0.67 and 0.90.
 Isooer could not be identified.

                                                             ~             (continued)
                                           26

-------
                                                                                          TABLE 12.  (continued)
Aliquot
Sample Air-dry wt. (g) TCDD
No. or Vol. (L) Isomer
8367-9 1-1576X-IWW



8367-86-1576X-INDD
8367-93- 1576X-IWWN







8367-70-1576X-H1N
-






8367-65-1576X-B5 -

8367-66-1576X-B5D

8367-67-1576X-B5N

_
"•
_
~

- • -
1.0 L 2,3,7,8-
1,3.6,8-
1,3,7,9-
1,2.7,8-
1.0 L 2,3,7,8-
1.0 L 2,3,7,8-
1,3.6,8-
1,3,7,9-
1.2.3.7/
1,2.3,8-
1,2,3,4-
1,2,7,8-
1,2,8,9-
.10.01 g~ 2,3,7,8-
1,3,6,8-
1,3,7,9-
1.2.3.7/
1,2,3,8-
1,2,3,4-
1,2,7,8-
1,2,8,9-
10.00 g 2,3,7,8-
1.3,7,9-
9.85 g 2,3,7,8-
1,3,7,9-
10.00 g 2,3,7.8-
1,3,6,8-
— 1,3,7,9-
1.2.3.7/
1.2,3,8-
. — 1,2,3,4-
1,2,7,8-
1,2,8,9-
HRCC/HRMS Analysis
TCDD (ppt
Retention tine or ppq) Instr. Relative Ion Abundance Ratios
TCDD lJC12-2,3,7,8 Meas. DL ID Date Time
22:00
17:03
18:14
24:30
21:50
21:39
16:47
17:57
21:58

21:11
24:08
29:35
21:38
16:45
17:55
21:58

22:10
24:09
29:36
21:47
18:05
21:44
18:03
21:47.
16:55
18:05
22:07

22:19 ~
24:16
29:45
21:57 534 - MS50 09/23/85 15:01
246
222
54.7 -
21:48 1,430 - MS50 09/23/85 15:31
21:38 530 - MS50 09/24/85 11:19
582
690
940

1,180
1,790
691
21:37 " 30.3 - MS50 09/24/85 12:55
29 .0 -
51.1 -
125
. _
118
252 - —
100
21:45 18.2 - MS50 -09/24/85 13:27
8.5 -
21:43 15.1 - MS50 09/24/85 13:58
4.2 -
21:46 12.9 - MS50 09/24/85 14:29
ND 9.2
15.2 -
61.8 -

— 54.1 - " —
147
63.9 -
320/322 332/334(15)
0.87 0.83
0.71
0.78
0.70
0.80 0.76
0.76 0.72
0.82
0.80
0.72

0.79
0.82
0.77
0.85 0.78
0.63
0.87
0.81

0.69
0.81
0.85
0.87 0.78
0.85
0.67 0.80
0.87
0.86 0.73
0.59
0.67
0.77

0.90
0.75
0.83
332/334(RS) \ Rec. m/z 259
0.81 77 72:1
47:1
' 36:1
7:1
0.79 29 49:1
0.73 71 23:1
42:1
43:1
49:1

50:1
72:1
22:1
0.77 56 12:1
15:1
24:1
46:1

38:1
73:1
20:1
0.75 73 12:1
9.4:1
0.72 85 18:1
6.3:1
0.71 48 9:1
11:1
14:1
48:1

35:1
73:1
26:1
S/N
o/z 322 ra/z 334(IS) Comments
> 62:1 > 63:1
48:1
32:1
16:1
63:1 > 33:1 Low recovery.
49:1 > 63:1
40:1
41:1
49:1

64:1
63:1
20:1
15:1 > 63:1
26:1
33:1
63:1

58:1
> 63:1
23:1
42:1 > 63:1
25:1
31:1 > 63:1
12:1
16:1 > 63:1
15:1 Ratio unacceptable.
20:1
63:1

58:1
63:1
22:1
.Aqueous cample data  reported  as ppq and  soil sample data presented as ppt.
Criteria for positive  identification  require that the ion ratios fall between 0.67 and 0.90.
 Isomer could not  be  identified.

                       _•   -                              -                (continued)
                                        -27

-------
                                                                                           TABLE  12.   (continued)
Aliquot
Sample Air-dry wt. (g)
No. or Vol. (L)
8367-68- 1576X-H1

8367-69-1576x-HlD


8367-71-1576x-Bl



8367-72-1576x-BlD



8367-73-1576x-BlN
—







8367-74-1576x-H3





8367-77-1576x-FA



TCDD
Isomer
9.67 g 2,3
1
10.00 g 2
1
1
1.02 g 2
1
1
c
1.05 g 2
1
1
c
1.03 g 2
1
1
c
1
1
1
1
1
1.15 g 2
1
1
c
1
c
9.94 g 2
1
1
c
,3
,3
,3
,3
,3
,3
,3

.3
,3
,3

,3
,3
,3

,2
,2
,2
,2
,2
,3
,3
,3

,2

,3
,3
,3
,4
,6
,7
,6
,7
,7
,6
,7

,7
,6
,7

,7
,6
,7

,3
,3
,3
,7
,8
,7
,6
,7

,7

,7
,6
,7
,7-
,8-
.8-
,8-
,9-
,8-
,8-
,9-

,8-
,8-
.9-

,8-
,8-
,9-

,6/
,8-
,4-
,8-
,9-
,8-
,8-
,9-

,8-

,8-
,8-
,9-
TCDD (ppt
Retention time or ppq) Instr
TCDD
21:
42
16:48
21:33
16:
17:
21:
16:
17:
19:
21:
16:
17:
19:
21:
16:
17:
19:
22:

22:
24:
29:
21:
16:
18:
19:
24:
26:
21:
16:
17:
42
52
37
44
54
04
35
42
53
02
41
48
59
09
01

15
12
41
44
49
00
10
58
03
36
42
53
19:02
lJC12-2,3,7,8 Meas. DL ID
21:40 34.3 - MS50
4.5 -
21:31 36.6 - MS50
5.2 -
10.0 -
21:35 937 - MS50
160
312
50.6 -
21:33 785 - MS50
201
308
ND 28.9
21:41 1,280 - MS50
333
635
52
518

695
1,170
463
21:42 2,020 - MS50
164 - -
237
70.6 -
31.7 -
27.3 -
21:34 1,720 - MS50
1,880
1,750
1,250
HRGC/HRMS Analysis
Relative Ion Abundance Ratios
Date Time 320/322 332/334(15) 332/334(RSj 1 Rec.
09/24/85 15:02 0.
0.
09/24/85 15:32 0.
0.
0.
09/25/85 10:01 0.
0.
0.
0.
09/25/85 10:30 0.
0.
0.
0.
09/25/85 11:27 0.
0.
0.
0.
0.

0.
0.
0.
09/25/85 13:00 0.
0.
0.
0.
0.
0.
09/25/85 13:30 0.
0.
0.
0.
82 0.67 0.75 73
67
70 0.74 0-.81 46
69
88
83 0.83 0.83 95
85
84
69
78 0.85 0.80 75
87
82
65
77 0.83 0.81 80
77
85
81
72

78
82
83
81 0.81 0.81 79
86
81
74
92
68
80 0.82 0.82 4
83
81
80
m/z 259
29:1
11:1
39:1
11:1
15:1
97:1
34:1
44:1
7.3:1
73:1
36:1
40:1
6.5:1
97:1
47:1
79:1
6.7:1
57:1

51:1
87:1
23:1
> 145:1
"22:1
24:1
9:1
2.5:1
3:1
67:1
109:1
94:1
60:1
S/N
ra/z 322 m/z 334(IS) Comments
63:1 > 63:1
14:1
31:1 > 54:1
5:1
10:1
63:1 > 63:1
14:1
24:1
4:1
63:1 > 63:1
22:1
28:1
3:1 Ratio unacceptable.
> 63:1
23:1
35:1
4:1
27:1

29:1
63:1
29:1
> 63:1 > 63:1
> 6:1
> 10:1
> 3:1
6:1
6:1
28:1 6:1 Lou recovery.
38:1
34:1
22:1
.Aqueous sample data reported as "ppq and soil sample  data presented as ppt.
 Criteria for positive identification require that  the  ion  ratios  fall between 0.67 and 0.90.
 Isomer could not be identified.

                                                                           (continued)
                                           28

-------
                                                                                           TABLE  12.   (continued)
Aliquot
Sample ' Air-dry we. (g) TCOD
No. or Vol. (L) Isomer
8367-77- 15 76X-FA c
(concluded) c
c
1,2,3,7/
1,2,3,8-
1,2,3,4-
c
1,2,7,8-
c
c
c
c
c
c
8367-96- 1576X-H2W 1.0 L_ 2,3,7,8-

1,3,6,8-
1,3,7,9-
c
c
c
1,2,7,8-
c
c
c
c
8367-83-1576X-DWD 1.0 L 2,3,7,8-
1,3,6,8-
1,3,7,9-
c
8367-82- 1576X-DW 1.0 L 2,3,7,8-
1,3,6,8-
1,3,7,9-
TCDD (ppt
Reteation tine or ppq) Instr
TCDD
20:00
20.43
21:25
21:54

22:08
22:35
24:05
24:49
25:23
25:55
27:18
28:07
29:41
21:42

16:47
17:58
19:08
20:06
20:49
24:13
24:55
26:01
27:25
28:13
21:11
16:25
17:34
23:39
21:10
16:22
17:32
lJCl2-2,3,7,8 Meas. DL ID
1,220 - MS50
274
109
675

4,640
879
720
3,460
155
3,430
441
2,920
ND 58
21:41 NC - MS50

NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
21:10 265 - MS50
ND 167
125
50.7 -
21:09 300 - MS50
140
106
HRGC/HRMS Analysis
Relative Ion Abundance Ratios
Date Time 320/322 332/334(15)
09/25/85 13:30 0.84
0.75
0.77
0.80

0.81
0.88
0.88
0.80
0.75
0.78
0.79
0.68
0.57
09/25/85 15:18 0.86 8.3

0.81
0.84
0.79
0.98
0.68
0.75
0.80
0.72
0.82
0.81
09/26/85 09:56 0.79 0.73
0.91
0.71
0.71
09/26/85 10:26 0.72 0.70
0.81
0.96
332/334(RS) X Rec. m/z 259
44:1
16:1
6:1
32:1

146:1
31:1
23:1
80:1
7:1
86:1
14:1
68:1
7:1
0.82 ND > 154:1

18:1
12:1
20:1
6:1
3:1
3:1
37:1
63:1
9:1
30:1
0.70 42 18:1
18:1
14:1
3.2:1
0.74 16 10:1
6.3:1
3.8:1
S/N
m/z 322 m/z 334(IS) Cooaents
15:1
5:
2.5:
11:

63:
14:
15:
56:1
5:1
63:1
9:1
46:1
4:1 Ratio unacceptable.
> 65:1 4:1 332/334(13) Ratio unacceptable; no amount
computations performed.
55:1
40:1
63:1
20:1 -
11:1
3:1
35:1
63:1
8:1
25:1
31:1 47:1
24:1 - Ratio unacceptable.
21:1
7:1
21:1 22:1 Low recovery. Rerun.
12:1
11:1
^Aqueous sample~data reported as ppq and soil sample data presented as ppt.
Criteria for positive identification require that the ion ratios fall between 0.67 and 0.90.
 Isomer could not be identified.

                                                                           (continued)
                                               29

-------
                                                                                           TABLE 12.  (continued)
Aliquot
Sample Air-dry wt. (g)
No. or Vol. (L)
8367-88-1576X-EWW 1.0 L



8367-95-1576X-H2WD 1.0 L















8367-96-1576X-H2WN 1.0 L

-



—



-

•
-

	
TCDD
Isomer
2,3.7,8-
1,3,6,8-
1,3,7,9-
1,2,7,8-
2,3,7,8-

1,3,6,8-
1,3,7,9-
c
c
c
c
1,2,7,8-
c
c
c
c
c
c
c
2,3,7,8-

1,3,6,8-
1,3,7,9-
c
c
c
c
1,2,7,8-
c
c
c
c
c
c
c
Retention time
TCDD '•JC12-2,3,7I8
21:18 21:15
16:28
17:38
23:44
21:13 21:12

16:25
17:35
18:43
19:38
20:20
22:11
23:39
24:23
25:27
26:47
27:37
29:34
30:12
31:07
21:14 21:13

16:26
17:35
18:44
19:41
20:22
21:13
23:40
24:24
25:30
26:51
27:39
29:36
30:14
31:12
TCDD (ppt
or ppq) Instr.
Meas. DL ID
1,030 - MS50
119
221
119
NC - MS50

NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC - MS50

NC
NC -
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
HRCC/HRMS Analysis
Relative Ion Abundance Ratios
Date Time 320/322 332/334(15) 332/334(RS)
09/26/85 12:32 0.76 0.79 0.75
0.71
0.68
0.76
09/26/85 13:05 0.93 13.3 0.67

0.77
0.82
0.81
0.87
0.79
0.84
0.70
0.76
0.76
0.79
0.73
0.71
0.81
0.58
09/26/85 14:21 0.87 9.47 0.87

0.79
0.78
0.78
0.89
0.63
0.59
0.79
0.73
0.75
0.76
0.70
0.77
0.74
0.33

% Rec. m/z 259
66 73:1
12:1
17:1
6:1
NC > 146:1

76:1
42:1
46:1
15:1
9:1
5:1
5:1
54:1
79:1
11:1
36:1
7:1
20:1
4:1
NC > 145:1

40:1
22:1
34:1
11:1
7:1
4:1
5:1
58:1
89:1
11:1
39:1
9:1
23:1
3:1
S/N
m/z 322 m/z -334(15) Comments
> 63:1 > 63:1
> 10:1
> 15:1
31:1
> 63:1 32:1 332/334(15) Ratio unacceptable; no amount
computations performed.
> 26:1
> 14:1
> 17:1
> 5:1
> 4:1
> 2:1
3.5:1
39:1
63:1
8:1
25:1
5:1
15:1
2.5:1
> 63:1 3:1 332/334(15) Ratio unacceptable; no amount
computations performed.
> 23:1
> 15:1
> 21:1
> 6:1
> 5:1 - Ratio unacceptable.
> 6:1 - Ratio unacceptable.
> 3:1
> 41:1
> 69:1
> 10:1
> 28:1
> 6:1
> 20:1
> 25:1 - Ratio unacceptable.
^Aqueous-sample data reported as ppq and soil sample data presented as  ppt.
 Criteria for positive identification require that the ion ratios  fall  between  0.67  and 0.90.
clsooer could not be identified.

                                                                           (continued)
                                             30

-------
                                                                                           TABLE 12.   (continued)
Aliquot
Sample Air-dry wt. (g)
No. or Vol. (L)
8367-75-1576X-H3D 1.16 g



8367-76-1576-X-H3N 1.14 g








8367-78-1576X-FAD 10.04 g









-





8367-99-1576X-FAN 9.93 g







"
2
1
1
c
2
1
1
c
1
1
1
1
1
2
1
1
c
c
c
1
1
1
c
c
c
. c
c
c
c
2
1
1
c
c
c
1
1
1
TCDD
I some r
.3
,3
,3

,3
,3
,3

,2
,2
,2
,2
,2
,3
,3
,3



,2
,2
,2







,3
.3
.3



,2
,2
,2
,7
,6
,7

,7
,6
,7

,3
,3
.3
,7
,8
,7
,6
,7



,3
,3
,3







,7
,6
,7



,3
,3
,3
.8-
.8-
.9-

,8-
,8-
,9-

,77
,8-
,*-
,8-
,9-
,8-
,8-
,9-



,11
,8-
,4-







,8-
.8-
,9-



,7/
,8-
,4-
TCDD (ppt
Retention time or ppq) Instr
TCDD
21:20
16:31
17:41
18:49
21:39
16:45
17:56
19:07
21:58

22:11
24:10
29:36
21:33
16:41
17:51
19:01
19:57
20:41
21:52

22:05
22:33
24:01
24:45
25:51
27:14
28:03
30:01
21:39
16:45
17:56
19:06
20:01
20:47
21:58

22:11
lJCl2-2,3,7,8 Meas. DL ID
21:19 2,260 - MS50
116
163

21:39 1,800 - MS50
383
367
ND 49.5
825

855
2,330
952
21:30 1,020 - HS50
926
747
610
557
146
286

2,260
MD 329
356
3,520
3,680
558
3,120
3,270
21:38 1,160 - MS50
1,390
1,160
881
888
194
423

3,620
HT.CC/HRMS Analysis
Relative Ion Abundance Ratios
Date Time 320/322 332/334(13)
09/27/85 15:31 0
0
0

09/30/85 09:57 0
0
0
1
0

0
0
0
09/30/85 10:29 0
0
0
0
0
0
0

0
0
0
0
0
0
0
0
09/30/85 13:00 0
0
0
0
0
0
0

0
.79 0.88
.90
.67

.80 0.79
.80
.81
.00
.81

.83
.85
.77
.73 0.71
.77
.80
.87
.89
.71
.67

.79
.91
.74
.78
.79
.83
.76
.79
.80 0.84
.78
.79
.85
.74
.76
•80

.86
332/334(RS) I Rec. ra/z 259
0.80 99 > 145:1
14:1
20:1
6:1
0.80 86 > 143:1
44:1
41:1
5:1
79:1

60:1
145:1
46:1
0.88 6.7 45:1
55:1
42:1
32:1
23:1
8.7:1
17:1

75:1
16:1
13:1
92:1
97:1
17:1
80:1
, 83:1
0.88 5 56:1
94:1
80:1
60:1
35:1
13:1
29:1
131:1
32:1
S/H
m/z 322 m/z 334(IS) Comments
> 63:
> 5:
> 7:
> 2.5
> 63:
> 15:
> 17:
> 2.5:
> 29:

> 24:
63:
23:
32:
37:
25:
22:
14:
6:
10:

63:
11:
8:
56:
63:
10:
48:
47:
25:
40:
28:
18:
17:
5:
10:
63:
10:
I > 63:1
1
1
:1
1 > 63:1
I
1
1 Ratio unacceptable.
1

\
1
1
1 9:1 Recovery low.
I
1
1
1
1
I

1
1 Ratio unacceptable.
1
1
1
1
1
1
1 6:1 Low recovery.
1
1
1
1
1
1
1
1
.Aqueous sample data reported as ppq and soil sample data presented as ppt.
Criteria for positive identification require that the ion ratios fall between 0.67 and 0.90.
 IsoBer could not be identified..

                                                                           (continued)
                                        -   31

-------
TABLE 12. (concluded)
Aliquot
Sample Air-dry wt. (g) TCDD
No. or Vol. (L) Isomer
8367-99-1576X-FAN
(concluded)






8367-100-1576X-DWD 1.0 L


8367-102-1576X-IND 500 mL
8367-101-1576X-EWD 1.0 mL


8367-103-1576X-IND 500 mL


8367-104-1576X-H2W 430 mL



~



8367- 105- 1576X-H2W 430 mL






-
c
1,2,7,8-
c
c
c
c
c
c
2,3,7,8*
1,3,6,8-
1,3,7,9-
2,3,7,8-
2,3,7,8-
1,3,6,8-
1,3,7,9-
2,3,7,8-
1,3,6,8-
1,3,7,9-
2,3,7,8-
1,3,6,8-
1,3,7,9-
c
c
c
c
c
2,3,7,8-
1,3,6,8-
1,3,7,9-
c
c
c
c
c
TCDD (ppt
Retention time or ppq) Instr
TCDD
22:38
24:08
24:52
25:58
27:20
28:10
30:09
30:47
21:04
16:19
17:26
21:02
20:58
16:14
17:23
20:57
16:17
17:23
21:00
16:18
17:27
18:32
24:06
25:09
27:16
29:49
20:56
16:15
17:22
18:30
24:03
25:07
24:17
29:45
1JCl2-2,3,7,8 Meas. DL ID
562 - MS50
516
4,310
4,530
ND 632
3,980
4,080
1,170
21:01 246 - MS50
637
489 -
21:03 604 - MS50
20:55 1,050 - MS50
157
384
20:56 628 - MS50
ND 45
87
20:59 27,100 - MS50
ND 71
164 -
391 -
ND 427
531 -
575 -
313 -
20:56 28,100 - MS50
224 -
302 -
453 -
564 -
730 -
518 -
347 -
HRCC/HRMS Analysis
Relative Ion Abundance Ratios
Date Time 320/322 332/334(13) 332/334(RS)
09/30/85 13:00 0.81
0.82
0.75
0.73
0.95
0.77
0.88
0.71
10/03/85 11:32 0.86 0.82 0.74
0.77
0.80
10/03/85 12:58 0.71 0.87 0.85
10/03/85 13:28 0.69 0.82 0.90
0.81

10/03/85 14:01 0.71 0.75 0.78
0.52
0.69
10/03/85 14:33 0.74 0.84 0.74
1.05
0.68
0.85
0.65
0.70
0.81
0.86
10/03/85 15:17 0.80 0.84 0.71
0.85
0.88
0.73
0.80
0.81
0.80
0.79

% Rec. m/z 259
5 32:1
20:1
127:1
146:1
20:1
24:1
122:1
26:1
68.5 21:1
73:1
51:1
59.5 16:1
80 72:1
17:1
28:1
57 18:1
6:1
8:1
78 > 145:1
9:1
16:1
30:1
19:1
32:1
28:1
18:1
96 > 145:1
16:1
17:1
30:1
20:1
33:1
23:1
13:1
S/N
m/z 322 m/z 334(IS) Comments
10:1
10:1
54:1
63:1
7:1 Ratio unacceptable.
46:1
44:1
14:1
18:1 > 63:1
63:1
47:1
31:1 > 63:1
63:1 > 63:1
13:1
26:1
42:1 > 63:1
13:1
17:1
> 63:1 > 63:1
18:1 Ratio unacceptable.
31:1
55:1
21:1 Ratio unacceptable.
32:1
26:1
27:1
> 63:1 > 63:1
19:1
21:1
34:1
14:1
31:1
14:1
10:1
.Aqueous sample data reported as ppq and soil sample data presented as ppt.
 Criteria for positive identification require that the  ion  ratios  fall between 0.67 and 0.90.
 Isomer could not be identified.
                                               32

-------
                                        TABU I).   ACCUUCT  AND HKCtStON OT THE HUGC/Him AMLTSIS TO* 2,3,7,8-TCDO
                                                            FROM UBORATOBT  AQUEOUS HATRIK SPIKES	
U>
CO
2.3,7,8-Troo 2.3,7,8-Tcno
Saaple •*trl« Spike level (ppq) Delected (ppq)
Dlatilled water (DW)



Efflmnt traatewater (CW)




Influeot matevater (IW)




Indnitrlil waatevater (IHD)




Industrial vaatevater (IHD)


Soil extract (H2H)



ISO
250
250

1,000
1,000
1,000


SOO
500
SOO


SOO
SOO
SOO


-
-

.
-





Average cone
RPR



Average cone
RPR



Average cone
RPR



Average cone
RPR


Average cone
RPDD


Average cone.
RPD
2)4
26S
2*6
. 2*8
12.5
1,090, 1,030
1,010
1,050
. 1,050
7.6
53*
SOS
S30
. 524
5.0
1,2*0
1,520
1,430
. 1,410
16
604
628
. 616
3.9
27,100
28,100
27,600
3.6
2,3.7,8-TCOO
Recovery (X)
•3.6
106
103
Average ree. 101
RPR 9.3
109, 10.1
101
IDS
Average rec. 105
RPR 7.6
107
102
106
Average rec. IDS
RPR 4.8
2S8
304
286
Average rec. 283
RPD 16
.
-

.
-


"C.,-2
AbaolMte



Average rec.
RPR



Average rec.
RPR



Average ree.
RPR



Average rec.
RPR


Average rec.
RPD


Average rec.
RPD
,3,7,8-TCOO
recovery (X)
•2
42
69
64
6]
61, 66
91
80
75
40
77
75
71
74
8.1
23
20
29
24
38
60
57
58
5.2
78
96
87
25
               Relative percent  raage (calculated  from the difference of the high and low values divided by the average of all  value*  and
              .•ulllplled by 100 percent).
               Relative percent  difference.

-------
                  TABLE  14.  PRECISION OF THE  IIRGC/IIRNS ANALYSIS FOR 2,3,7,8-TCDD
                                    OF SOIL AND FLY  ASH SAMPLES
Saaple •atrin	

B25-Plazza Road (BS)
Hyde Park 001 (III)
B52-Shen«ndoah (Bl)
Hyde Park 003 (111)
Fly ash - RRAI

Endogenous
2,3,7,8-TCI)»
level (Pl>l)a
50


Average
RPR°
70


Average
RPR
360


Average
RPR
1,700


Average
RPR
.


Average
RPR


2,3,7,8-TCDD
Detected (ppt)
18.2
15.1
12.9
rone. 15.4
34
34.3
36.6
30.3
cone . 33 . 7
19
937
785
1,280
cone. 1,000
50
2,020
2,260
1,800
cone. 2,0.10
23
1,720
1,020
1,160
eonc. 1,300
54
•

>3C,2-2
Absolute



Average rec.
RPR



Average rec.
RPR



Average rec.
RPR



Average rec.
RPR



Average rec.
RPR


,3,7,8-TCDD
recovery (J.)
73
85
48
69
54
73
46
56
58
47
95
75
80
83
24
79
99
86
88
23
4
7
5
5.3
57

"Estimated level of endogenous 2,3,7,8-TCDD  reported  to NRI  by  Dr.  W.  Beckert  in letters dated
bApril 19, 1985, and August .10,  1985.
 Relative percent range (calculated  frno  the difference of the  high and low values,  divided by
 the average ol all values, and 1111111111 led by  100 percent.

-------
                     TABLE IS.  ACOimCT Of THE HRCC/ICWO IgtllOO FOB THE DCTtRHIMTIOH Of TCTO ISCTBO SPIKED HTTP AQUEOUS HATHICES
Efflueat vaatc
fCDD
analyte
1.3.6.8
1,3,7.9
1.2.3.7/1,2,3,8
1,2.3.4
1.2.7,8
1.2,8.9
2.3.7.8
"Clt-2,3. 7,8
Spike
(PI)
1.840
840
1,680
2,440
3,080
1,200
1,000
500
Heaaiired
(PR)
502
766
1,860
1.840
3,430
1.330
1,010
455
water 	
Recovery
U)
27
91
110
75
III
III
101
91
Dlatllled water
Spike
(PI)
460
210
420
610
770
300
250
500
Heaanred
(PS)
512
395
403
616
840
328
234
410
Recovery
(1)
III
190
96
101
110
110
94
82
Inl
Spite
(PI)
920
420
840
1,220
1.540
600
500
500
TlMnt vaatevater
Heamred
(PI)
582
690
940
1.180
1.790
691
530
355
Recovery
(I)
63
164
112
97
116
IIS
106
71
Indnatrlal vaatevater
Spike
(PI)
920
420
840
1.220
1,540
600
500
500
(featured
(PI)
HD'
HD
HD
RD
586
HD
904"
100
Recovery
(t)
0
0
0
0
38
0
181
20
           jHot detected.                                   	
            Meaaured value corrected for eedogenoaa 2,3,7,8-TCDD content (averaged 616 pi/L).
Oi

-------
          ARS36B(V),  I
                        TABU  16.  ACCURACY OP THE HHCC/HRM3 METHOD Km THE PETERrllHATIO*! Of TCTO ISOHERS SPIKED IHTO SOU HATH I Ct 3
Hyde Park 001
TCOO
•oalfte
1,3,6,8
I.3.M
1,2,3,7/1,2,3.8
1,2,3.4
1,2,7.8
1,2.8.9
2.3,7.8
l3Clt-2.3.7.8
Spike
(PI)
130
60
120
170
220
84
-
500
Heasored
(PI)
29.0
Sl.l
125
118
252
100
30.3
280
(HI)
Recovery
(M
22
86
106
69
117
119
-
56
B25-Piazza Road
Spike
_j£il
92
42
84
120
ISO
60
-
500
Measured
(PI)
m> (9.2)*
IS. 2
61.8
54.1
147
63.9
12.9
240
JMJ 	
Recovery
(1)
0
36
74
44
95
107
-
48
B52-Sbenandoah
Spike
(pi)
660
300
600
880
1,110
430
-
500
Measured
(PI)
333
63S
SIB
695
1.170
463
1.280
400
m 	
Recovery
(t)
SO
210
87
79
106
108
-
80
HT«
iplkV
(Pi)
1.560
710
1.430
2,070
2.620
1.020
-
500
le Park 003
Heaaured
(PI)
383
367
825
855
2,330
9S2
1,800
430
(H3)
Recovery
24
SI
58
41
89
93
-
86

to
             *ND - not detected.  The val«e  U parentheses reflects the cstiuted detection limit.

-------
              TABLE  17.  FORTIFIED FIELD BLANK RESULTS
Sample
Ho.
8367-62- 1576X-FFV3
8367-64-1576X-FFSB
8367-63- 15 76X-FFSA
8367-61-1576X-FFWA
8367-81-1576X-FFB
8367-80-1576X-FFA
8367-97-1576X-FFA
8367-98- 1576X-FFB
Aliquot
Air-dry wt. (g)
or Vol. (L)
1.0 L
10 g
10 g
1.0 L
10.01 g
10.01 g
1.0 L
1.0 L
Retention tine
Native
23:38
23:38
23:40
23:40
22:17
21:37
21:37
21:43
13C
23:35
23:38
23:38
23:39
22:15
21:35
21:37
21:43
Instr.
ID
MS50
MS50
MS50
MS50
MS50
MS50
MS50
MS50
Relative Ion Abundance Ratios
Date
09/11/85
09/11/85
09/12/85
09/12/85
09/20/85
09/20/85
09/20/85
09/20/85
Time
14:08
14:42
14:47
15:14
13:33
14:09
09:10
09:24
320/322
0.78
0.79
0.77
0.80
0.88
0.86
0.82
0.80
332/334(15)
-
-
0.78
0.76
-
0.73
0.83
-
332/334(RS)
0.71
0.80
-
-
0.74
-
-
0.82
% Rec.
65
68
71
79
29
83
50
48
m/z 259
> 145:1
> 145:1
145:1
145:1
144:1
145:1
23:1
145:1
S/N
ra/z 322
> 63:1
> 63:1
> 63:1
> 63:1
> 63:1
> 63:1
> 63:1
> 63:1

m/z 334(IS)
> 63:1
> 63:1
> 63:1
> 63:1
> 63:1
> 63:1
> 63:1
> 63:1
Comments




Low recovery



37

-------
HRGC and Mass Resolution

     Table  11 presents  a summary of all chromatographic and mass resolution
checks  completed during the  final method evaluation.  As  per  the protocol
requirements the required  mass resolution was demonstiated as the first and
last  quality control  requirements  for  each  day.   The  column  performance
check mixture was also  analyzed before the  first  sample  analysis and after
the  final  sample analysis  each day as  a QC measure to  assure  that speci-
ficity  for  2,3,7,8-TCDD was  maintained.   The mass  measurement  accuracy at
m/z  330.979 is  also included  in this  table, as  it was verified  on a daily
basis prior to any sample analyses.

Sample Analysis

     The results from the  analysis of the aqueous and soil samples are pro-
vided in Table  12.   The data  in Table 12 are presented in the format speci-
fied  as  Form B-l  in the protocol  reporting requirement.   The data are re-
corded  in   the  chronological  order  that they  were obtained by HRGC/HKMS.

     As  indicated in Table  12, several samples required  reanalysis due to
low  recovery of  the internal  standard, unacceptable ion ratios for 320/322,
or  the  result of  interferences at the internal standard.  Two  of the dis-
tilled water  samples demonstrated  responses for the  characteristic ions at
m/z  259,  320, and  322   for  2,3,7,8-TCDD.   However, the  ion ratio  for the
native  2,3,7,8-TCDD in  one  replicate  and  the ion  ratio  for the internal
standard in another required  that both samples be reanalyzed.  Although both
samples met all  the  qualitative criteria,  recoveries were noted to be low
(< 20 percent) for  one  of  the  samples and complete reanalysis of the repli-
cate was required.

     Significant  problems  were encountered  with  the aqueous  soil extract,
H2W,  and  the fly  ash sample.  The problems with  the  soil extract resulted
from  an  interference at m/z 332 that coeluted with  the  internal standard,
13C12-2,3,7,8-TCDD.  No accurate quantitative measurements could be achieved
for  TCDD responses  observed for this sample.  The original sample contained
a  large  amount  of  suspended particulate in each of  the three  replicates.
Problems with the extraction were  noted with the first portion of methylene
chloride.   A large  amount  of particulate matter was  noted at the interface
of  the  aqueous  and organic phases.  Hence,  the interference  at  m/z 332 and
TCDD responses observed  in these replicates were probably due to direct ex-
traction of the  suspended  soil particulate  rather  than the  actual water-
soluble TCDD.

     The remaining  aqueous  sample  for H2W was centrifuged for 10 min at ap-
proximately 2,000  rpm,  and the aqueous phase was  decanted from the settled
particulate.  The resulting aqueous sample was divided into duplicate 430-mL
samples  and each  was analyzed according to  the protocol.   The HRGC/HRMS
analysis was  successful  for  both replicates with absolute  recoveries of
78 percent and 96 percent of  the internal standard.

     The triplicate analyses of  the  fly ash  sample resulted  in absolute
recoveries  less  than 10 percent for the internal  standard in each aliquot

                                      38

-------
analyzed.  These  low recoveries may be associated with the total fixed car-
bon  content  of the fly ash material.  Previous work in this laboratory with
fly ash from coal-fired power plants has demonstrated low recoveries of ana-
lytes from materials with high carbon content.4

     The only other sample for which successful analysis was not achieved as
specified  in the  protocol  on first analysis  was  the  industrial wastewater
(IND).  The  triplicate  analysis of the sample resulted in absolute internal
standard recoveries  of  23,  20, and 29 percent.  The criteria for successful
analysis for TCDD as discussed in the protocol require an absolute recovery
of 40 to 120 percent.  In addition to the observed low recoveries, the level
of 2,3,7,8-TCDD detected in the sample averaged 1,410 ppq as compared to the
500-ppq spike  level.   Two 500-mL aliquots of the unspiked industrial waste-
water sample were reanalyzed to determine the background  level  of 2,3,7,8-
TCDD.  The results of the duplicate analysis yielded an average 2,3,7,8-TCDD
concentration  of  approximately 620 ppq  and  the  absolute  recoveries  were
noted to be 60 percent and 57 percent.  The increase in absolute recovery of
the  internal  standard in  the unspiked sample by  approximately  a factor of
two  is possibly due to the preparation of samples one half the size of that
used for the  original analysis.  This suggests that the sample matrix has a
considerable impact on the effectiveness of the cleanup procedure.

     Table 13 provides a summary of the accuracy and precision of the analy-
ses of the five aqueous sample types for 2,3,7,8-TCDD.   Only the data points
from Table 12  that demonstrate compliance with all QC criteria (ion ratios,
absolute recovery of the internal standard, etc.) are included in Table 13.
These data demonstrate that the isotope dilution method of quantitation pro-
vides accurate  and precise  quantitation of 2,3,7,8-TCDD in the aqueous sam-
ples.  It should be noted that even when the absolute recovery of the 13Ci2~
2,3,7,8-TCDD internal standard varies by as much as 66 percent (RPR) for the
triplicate distilled  water  samples,  the accuracy  of the measurement of the
spiked 2,3,7,8-TCDD averaged 101 percent with less than 10 percent variabil-
ity.  Table 13  summarizes  data for both the spiked and unspiked aliquots of
industrial wastewater.   The high recovery noted  for the  2,3,7,8-TCDD value
in  the  spiked  samples is a  result  of the presence of  this  compound at ap-
proximately 620 ppq in the original matrix.

     Table 14  presents a  similar summary  for the five solid samples ana-
lyzed.  The precision of the measurements is not quite as good as noted for
the  aqueous  samples  and may  reflect the  difference  in adsorption  of the
endogenous 2,3,7,8-TCDD  and the  spiked internal  standard  to the matrices.

     Tables  15  and  16 provide data  dealing with  the accuracy of the HRGC/
HRMS  methods for the determination  of total TCDD  isomers in  aqueous and
solid samples.   In general,  the data support  the  use  of the internal stan-
dard  method  of  quantitation   for  all  but the  earliest   eluting isomers,
1,3,6,8- and 1,3,7,9-TCDD.   The accuracy for the additional isomers is very
good and more  consistent than is observed  for the solid samples.  This may
be  partially  due  to  the differences in  adsorption to  the soil particles.
                                      39

-------
Fortified Field Blanks

     As  part  of the overall quality  assurance/quality  control (QA/QC) pro-
gram identified  in the HRGC/HRMS protocol, the  analyst  is  required to ana-
lyze  fortified  field  blanks  to demonstrate  (a)  that  the extraction  and
cleanup  procedure  will provide recovery of the 2,3,7,8-TCDD within the cri-
teria of greater than 40 percent specified in the protocol and (b) that the
reagents are free  from contamination with TCDD isomers.

     Table  17  provides the results of the fortified field blanks run before
proceeding  with sample  analysis  and also  those  of an additional  set  of
blanks prepared  along with the actual samples.  The  analyses  of the forti-
fied field  blanks  at the outset of the study demonstrated that the recover-
ies of 2,3,7,8-TCDD and 1,2,3,4-TCDD ranged from 65  to  79  percent.  No de-
tectable  levels  of other TCDD isomers were found in this preliminary study.
The field fortification blanks analyzed with the actual samples resulted in
recoveries  of  29 percent and  83 percent.  More  importantly,  these analyses
demonstrated  some  interferences  arising  from  1,3,6,8- and  1,3,7,9-TCDD.
Previous  studies  involving  evaluation of  the  cleanup  procedure  indicated
that these isomers are associated with the acidic alumina cleanup.

     Figure 4  is  a plot  of  the  ratio of response of 1,3,6,8- and 1,3,7,9-
TCDD and the response of the recovery standard 13Cj2-l,2,3,4-TCDD versus the
time elapsed  since the  acidic alumina was cleaned and  activated  at 190°C.
The results of  the  analyses  of  the fortified field blanks  and the samples
not spiked  with  the 1,3,6,8- and 1,3,7,9-TCDD isomers are presented in Fig-
ure 4.   As  noted from this plot, these TCDD  isomers  were not initially de-
tected in the acidic  alumina  immediately following cleanup by Soxhlet ex-
traction.   The first  set  of fortified field  blanks  was taken through the
acidic alumina column 7 days  later.  Although response  was  observed at m/z
320 and  322 at the retention time for these isomers,  the ion ratios did not
indicate presence  of the compounds.  Since the  detectable  levels  were well
below 10 pg/g  of alumina, the sample analyses were initiated.   The data for
the fortified  field blanks and samples taken  through alumina  from 14 to 30
days from activation  indicate that the contamination  of the  1,3,6,8-  and
1,3,7,9-TCDD isomers apparently occurs over time using this particular oven.
The background contamination of 1,3,6,8- and  1,3,7,9-TCDD  isomers  has also
been recently addressed by the Center for Disease Control.5

Note Added in Proof

     A second  magnetic sector instrument  (built in  1976)  from a  different
manufacturer was tested  and  found to be incapable of  achieving sufficient
sensitivity at 10,000  resolving  power to be  used in  experiments  for this
study.
                                      40

-------
                         Fortified Field Blanks


                      [J Aqueous and Environmental Samp I
                             10             20
                    Time (DaysJ Elapsed from Cleanup and
                        Activation of Acidic Alumina  "~
Figure 3.
Background levels  of 1,3,6,8- and 1,3,7,9-TCDD observed
over the single-laboratory evaluation study.

                       41

-------
                                 REFERENCES
1.  U.S. Environmental Protection Agency,  "Dioxin Strategy," prepared  by the
    Office of Water Regulations  and Standards and the Office of Solid Waste
    and  Emergency  Response  in  conjunction  with  the  Dioxin Strategy  Task
    Force, Washington, D.C., November 28,  1983.

2.  L. R. Williams, Validation of Testing/Measurement Methods.
    EPA 600/X-83-060,  1983.

3.  GC Bulletin 793C,  Supelco Inc., Beliefonte,  Pennsylvania, 1983.

4.  C.  L.  Haile,  J.  S.  Stanley, T. Walker,  6.  R. Cobb, and B. A. Boomer,
    "Comprehensive Assessment  of the Specific Compounds Present in Combus-
    tion  Processes.   Volume 3.   National  Survey of Organic Emissions  from
    Coal-Fired  Utility  Boiler Plants,"  EPA-560/5-83-006,  September  1983.

5.  J.  S. Heller,  D. G.  Patterson,  L. R.  Alexander, D.  F.  Groce,  R.  P.
    O'Connor, and  C.  R.  Lapeza,  "Control  of Artifacts and  Contamination in
    the Development of a Dioxin  Analytical Program," presented at the  33rd
    Annual  Conference on  Mass Spectrometry and  Allied  Topics, May  26-31,
    1985, San Diego, California.
                                      42

-------
APPENDICES
    43

-------
                                   APPENDIX A

                         VALIDATED ANALYTICAL PROTOCOL
                            for the Determination of
          2,3,7,8-Tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD)  and  Total
            TCDDs in Soil/Sediment and Water by High-Resolution Gas
                Chromatography/High-Resolution Mass Spectrometry
                               September 10, 1985

     This analytical protocol has been written in the format used in the
Superfund program, as "Exhibit D" of a Statement of Work  which  in turn  is part
of an Invitation-for-Bid package under the Superfund Contract Laboratory Program.
The other exhibits of the Statement of Work, although cited  in  Exhibit  D, do
not pertain to this method evaluation study.

-------
                         EXHIBIT D
                     Analytical Methods

2,3,7,8-Tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD)  and Total
  TCDDs in Soil/Sediment and Water by High-Resolution Gas
      Chromatography/High-Resolution Mass Spectrometry

-------
                                   EXHIBIT D









Section                       Subject






   1               Scope and Application	    D-l




   2               Summary of Method	    D-l




   3               Definitions	    D-2




   A               Interferences	    D-3




   5               Safety	    D-3




   6               Apparatus and Equipment	    D-3




   7               Reagents and Standard Solutions	    D-6




   8               System Performance Criteria	    D-8




   9               Quality Control Procedures	    D-l3




  10               Sample Preservation and Handling ......    D-l3




  11               Sample Extraction	    D-14




  12               Analytical Procedures	    D-l7




  13               Calculations	    D-18

-------
1.   SCOPE AND APPLICATION

     1.1  This method provides procedures for the detection  and  quantitative
          measurement of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD; CAS
          Registry Number 1746-01-6; Storet number 3475)  at  concentrations of
          10 pg/g (10 parts per trillion) to 200 pg/g (200 parts per  trillion)
          in 10-g portions of soil and sediment and at 100 pg/L  (100  parts per
          quadrillion) to 2000 pg/L (2 parts per trillion) in 1-L samples of
          water.  The use of 1-g or 100-mL portions permits  measurements of
          concentrations up to 2,000 pg/g (2 parts per billion)  or 20 ng/L,
          respectively.  This method also allows the estimation  of quantities
          of total TCDD present in the sample.   Samples containing concentrations
          of 2,3,7,8-TCDD greater than 2 ppb or 20 ng/L must be  analyzed by a
          protocol designed for such concentration levels, with  an appropriate
          instrument calibration range.

     1.2  The minimum measurable concentration is estimated  to be 10  pg/g (10
          parts per trillion) for soil and sediment samples  and  100 pg/L for
          water samples, but this depends on kinds and concentrations of
          interfering compounds in the sample matrix.

     1.3  This method is designed for use by analysts who are experienced in
          the use of high-resolution gas chromatography/high-resolution mass
          spectroraetry.

     CAUTION:  TCDDs are extremely hazardous.  It is the laboratory's responsi-
               bility to ensure that safe handling procedures are employed.

2.   SUMMARY OF METHOD

     Five hundred pg of 13C,2~2,3,7,8-TCDD (internal standard) are added to a
     10-g portion of a soil/sediment sample (weighed to 3 significant figures)
     or a 1-L aqueous sample and the sample is  extracted with 200 to  250 mL
     benzene using a Soxhlet apparatus with a minimum of 3 cycles per hour or a
     continuous liquid-liquid extractor for 24  hours.  A separatory funnel and
     3 x 60 mL methylene chloride may also be used for aqueous samples.  After
     appropriate concentration and cleanup, 50  uL of tridecane are added to the
     extract.  Before HRGC-HRMS analysis, 500 pg of a recovery standard (  Cj2~
     1,2,3,4-TCDD) are added to the extract which is then concentrated to a
     final volume of 50 uL.  A 2-uL aliquot of the concentrated  extract is
     injected into a gas chromatograph with a capillary column interfaced to a
     high-resolution mass spectrometer capable of rapid multiple ion  monitoring
     at resolutions of at least 10,000 (10 percent valley).

     Identification of 2,3,7,8-TCDD is based on the detection of the  ions m/z
     319.897 and 321.894 at the same GC retention time and within -1  to +3
     seconds GC retention time of the internal standard masses of m/z 331.937
     and 333.934.  Confirmation of 2,3,7,8-TCDD (and of other TCDD isomers) is
     based on the ion m/z 258.930 which results from loss of COCL by  the parent
     ion.
                                      D-l

-------
3.   DEFINITIONS

     3.1  Concentration calibration solutions — solutions  containing  known
          amounts of the analyte (unlabeled  2,3,7,8-TCDD),  the  internal  standard
          13C12-2,3,7,8-TCDD and the recovery standard  13C,2-l,2,3,4-TCDD;
          they are used to determine instrument  response  of  the analyte
          relative to the internal standard  and  of the  internal standard
          relative to the recovery standard.

     3.2  Field blank — a portion of soil/sediment or  water uncontaminated with
          2,3,7,8-TCDD and/or other TCDDs.

     3.3  Rinsate — a portion of solvent used to rinse sampling equipment; the
          rinsate is analyzed to demonstrate that samples have  not  been  contami-
          nated during sampling.

     3.4  Internal standard —  ^Cj2~2,3,7,8-TCDD,  which  is  added to every
          sample (except the blanks described in Sections 4.2.1 and 4.2.3 of
          Exhibit E) and is present at  the same  concentration in every labora-
          tory method blank, quality control sample,  and  concentration calibra-
          tion solution.  It is added to the soil/sediment  or aqueous  sample
          before extraction and is used to measure  the  concentration of  each
          analyte.  Its concentration is measured in every  sample,  and percent
          recovery is determined using  an internal  standard  method.

     3.5  Recovery standard —   ^12-l»2,3,4-TCDD which is  added to every sample
          (except for the blanks discussed  in Sections  4.2.1.A.2 and 4.2.3.6,
          Exhibit E) extract just before HRGC-HRMS analysis.

     3.6  Laboratory method blank — this blank  is prepared  in  the  laboratory
          through performing all analytical  procedures  except addition of a
          sample aliquot to the extraction vessel.

     3.7  GC column performance check mixture — a  mixture containing  known
          amounts of selected standards; it  is used to  demonstrate  continued
          acceptable performance of the capillary column, i.e., separation
          (£ 25% valley) of 2,3,7,8-TCDD isomer  from all  other 21 TCDD isomers
          and to define the retention time window.

     3.8  Performance evaluation sample — a soil,  sediment  or  aqueous sample
          containing a known amount of  unlabeled 2,3,7,8-TCDD and/or other
          TCDDs.  It is distributed 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 ("blind") QC sample  in any
          sample batch submitted to a laboratory for  analysis.

     3.9  Relative response factor — response of the mass  spectrometer  to a
          known amount of an analyte relative to a  known  amount of  an  internal
          standard.

     3.10 Mass resolution check — standard  method used to  demonstrate static
          resolution of 10,000 minimum  (10Z  valley  definition).

                                     D-2

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4.   INTERFERENCES

     Chemicals which elute from tbe GC column within ^10 scans of the internal
     and/or recovery standard (m/z 331.937 and 333.934)  and which produce ions
     at any of the masses used to detect or quantify TCDD are  potential  inter-
     ferences.  Most frequently encountered potential interferences are  other
     sample components that are extracted along with TCDD,  e.g. PCBs, methoxy-
     biphenyls, chlorinated hydroxydiphenylethers,  benzylphenylethers, chlori-
     nated naphthalenes, DDE, DDT, etc.  The actual incidence  of interference
     by these chemicals depends also upon relative  concentrations, mass  spectro-
     metric resolution, and chromatographic conditions.   Because very low
     levels of TCDD must be measured,  the elimination of interferences is
     essential.  High-purity reagents  and solvents  must  be  used and all  equip-
     ment must be scrupulously cleaned.  Laboratory reagent blanks (Exhibit E,
     Quality Control, Section 4) must  be analyzed to demonstrate absence of
     contamination that would interfere with TCDD measurement.  Column chromato-
     graphic procedures are used to remove some coextracted sample components;
     these procedures must be performed carefully to minimize  loss of TCDD
     during attempts to increase its concentration  relative to other sample
     components.

5.   SAFETY
                          •

     The toxicity or carcinogenicity of each reagent used in this method has
     not been precisely defined; however, 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 for 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  identi-
     fied d~3) (page D-38).  2,3,7,8-TCDD has been identified as a suspected
     human or mammalian carcinogen.  The laboratory is responsible for ensuring
     that safe handling procedures are followed.

6.   APPARATUS AND EQUIPMENT

     6.1  High-Resolution Gas Chromatograph/High-Resolution Mass
          Spectrometer/Data System (HRGC/HRMS/DS)

          6.1.1  The GC must be equipped for temperature programming, and all
                 required accessories must be available, such  as syringes, gases,
                 and a capillary column.  The GC injection port must be  designed
                 for capillary columns.  The use of splitless  injection  tech-
                 niques is recommended.  On-column  injection techiques can be
                 used but this may severely reduce  column lifetime for non-
                 chemical ly bonded columns.  When using  the method in this
                 protocol, a 2-uL injection volume  is used consistently.  With
                 some GC injection ports, however,  1-uL  injections may produce
                 improved precision and chromatographic  separation.  A 1-uL
                                      D-3

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                 injection volume may be used if adequate  sensitivity  and
                 precision can be achieved.

          NOTE:  If 1 uL is used at all as injection volume,  the  injection
                 volumes for all extracts, blanks,  calibration  solutions and
                 the performance check sample must  be 1  uL.

          6.1.2  Gas Chromatograph-Mass Spectrometer Interface

                 The GC-MS interface may include enrichment  devices, such as
                 a glass jet separator or a  silicone membrane separator, or
                 the gas chromatograph can be directly coupled  to the  mass
                 spectrometer source.  The interface may include  a diverter
                 valve for shunting the column effluent  and  isolating  the mass
                 spectrometer source.  All components of the  interface should
                 be glass or glass-lined stainless  steel.  The  interface com-
                 ponents should be compatible with  300°C temperatures. The
                 GC/MS interface must be appropriately designed so that the
                 separation of 2,3,7,8-TCDD  from the other TCDD isomers which
                 is achieved in the gas chromatographic  column  is not  appreci-
                 ably degraded.  Cold spots  and/or  active  surfaces (adsorption
                 sites) in the GC/MS interface can  cause peak tailing  and peak
                 broadening.  It is recommended that the GC  column be  fitted
                 directly into the MS source.  Graphite  ferrules  should be
                 avoided in the GC injection area since  they may  adsorb TCDD.
                 Vespel* or equivalent ferrules are recommended.

          6.1.3  Mass Spectrometer

                 The static resolution of the instrument must be  maintained at
                 a minimum 10,000 (10 percent valley).  The  mass  spectrometer
                 must be operated in a selected ion monitoring  (SIM) mode with
                 total cycle time (including voltage reset time)  of one second
                 or less (Section 8.3.4.1).   At a minimum, the  following ions
                 which occur at these masses must be monitored:   m/z 258.930,
                 319.897, 321.894, 331.937 and 333.934.

          6.1.4  Data System

                 A dedicated hardware or data system is  employed  to control the
                 rapid multiple ion monitoring process and to acquire  the data.
                 Quantification data (peak areas or peak heights) and  SIM traces
                 (displays of intensities of each m/z being  monitored  as a
                 function of time) must be acquired during the  analyses.
                 Quantifications may be reported based upon  computer-generated
                 peak areas or upon measured peak heights  (chart  recording).

NOTE:  Detector zero setting must allow peak-to-peak measurement  of the noise
       on the base line.

     6.2  GC Columns


                                      D-4

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     For isomer-specific determinations of 2,3,7,8-TCDD,  the following  two
     fused silica capillary columns are recommended:   a  60-m SP-2330  column
     and a 50-tn CP-Sil 88 column.  However,  any  capillary column which
     separates 2,3,7,8-TCDD from all other TCDDs may  be  used for such
     analyses, but this separation must be demonstrated  and documented.
     Minimum acceptance criteria must be determined per  Section 8.1.  At
     the beginning of each 12-hour period (after mass resolution has  been
     demonstrated) during which sample extracts  or concentration calibra-
     tion solutions will be analyzed, column operating conditions must  be
     attained for 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  2 at the end of  this
     Exhibit.

6.3  Miscellaneous Equipment

     6.3.1  Nitrogen evaporation apparatus with  variable flow rate.

     6.3.2  Balance capable of accurately weighing to 0.01 g.

     6.3.3  Centrifuge capable of operating at 2,000  rpm.

     6.3.4  Water bath — equipped with concentric ring  cover and capable
            of being temperature-controlled within *2°C.

     6.3.5  Stainless steel spatulas or spoons.

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

     6.3.7  Glove box.

     6.3.8  Drying oven.

6.4  Glassware

     6.4.1  Soxhlet apparatus — all-glass,  Kontes 6730-02 or equivalent;
            90 mm x 35 mm glass thimble; 500-mL flask; condenser of appro-
            priate size.

     6.4.2  Kuderna-Danish apparatus — 500-mL evaporating flask, 10-mL
            graduated concentrator tubes with ground-glass stoppers,  and
            3-ball macro Snyder column (Kontes K-570001-0500, K-503000-
            0121 and K-569001-0219 or equivalent).

     6.4.3  Mini-vials — 1-mL borosilicate glass with conical-shaped
            reservoir and screw caps lined with  Teflon-faced silicone disks.

     6.4.4  Funnels — glass; appropriate size to accommodate filter
            paper used to filter jar extract (volume  of  approximately 170 mL)

     6.4.5  Separatory funnel — 2000 mL with Teflon  stopcock.

                                 D-5

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          6.A.6  Continuous liquid-liquid extractors equipped with Teflon or
                 glass connecting joints and stopcocks requiring no lubrication
                 (Hershberg-Wolf Extractor - Ace Glass Company Vineland,  NJ,
                 P/N 6841-10 or equivalent).

          6.4.7  Chromatographic columns for the silica and alumina chroma-
                 tography — 1 cm ID x 10 cm long and 1 cm ID x 30 cm long.

          6.4.8  Chromatography column for the Carbopak cleanup — disposable
                 5-mL graduated glass pipets, 7 mm ID.

          6.4.9  Desiccator.

          6.4.10 Glass rods.

          NOTE:  Reuse of glassware should be minimized to avoid the risk of
                 cross contamination.  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 then with
                 high-purity acetone and hexane.  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°C for 15 to 30 minutes. Volumetric glassware
                 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 the glassware is dry and  cool, rinse with hexane,
                 and store inverted or capped with solvent-rinsed aluminum foil
                 in a clean environment.

7.   REAGENTS AND STANDARD SOLUTIONS

     7.1  Column Chromatography Reagents

          7.1.1  Alumina, acidic — Extract the alumina in a Soxhlet with
                 methylene chloride for 6 hours (minimum of 3 cycles per  hour)
                 and activate it by heating in a foil-covered glass container
                 for 24 hours at 190°C.

          7.1.2  Silica gel — high-purity grade, type 60,  70-230 mesh; extract
                 the silica gel in a Soxhlet with methylene chloride for  6 hours
                 (minimum of 3 cycles per hour) and activate it by heating in a
                 foil-covered glass container for 24 hours at 130°C.

          7.1.3  Silica gel impregnated with 40 percent (by weight) g-ulfuric
                 acid — add two parts (by weight) concentrated sulfuric  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.

                                      D-6

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     7.1.A  Sulfuric acid,  concentrated — ACS grade,  specific  gravity  1.84.

     7.1.5  Graphitized carbon black (Carbopack C or equivalent),  surface
            of approximately 12 m^/g,  80/100 mesh — mix thoroughly  3.6
            grams Carbopak C and 16.4  grams Celite 545® in a 40-mL vial.
            Activate at 130° C for six hours.  Store in a desiccator.

     7.1.6  Celite 545®, reagent grade, or equivalent.

7.2  Membrane filters or filter paper  with pore size of <25 urn;  rinse with
     hexane before use.

7.3  Glass wool,  silanized — extract  with methylene chloride and  hexane
     and air-dry before use.

7.4  Desiccating Agents

     7.4.1  Sodium sulfate — granular, anhydrous; before use,  extract  it
            with methylene  chloride for 6 hours (minimum of 3 cycles per
            hour) and dry it for >4 hours in a shallow tray placed in an
            oven operated at 120°C.  Let it cool in a  desiccator.

     7.4.2  Potassium carbonate—anhydrous, granular;  use as such.

7.5  Solvents -- high purity, distilled in glass:  methylene chloride,
     toluene, benzene, cyclohexane, methanol, acetone,  hexane;  reagent
     grade:  tridecane.

7.6  Concentration calibration solutions (Table 1) —  five tridecane
     solutions containing unlabeled 2,3,7,8-TCDD and 13C,a~l»2,3,4-TCDD
     (recovery standard) at varying concentrations, and   C,9-2,3,7,8-TCDD
     (internal standard, CASRN 80494-19-5) at a constant concentration
     must be used to calibrate the instrument.  These  concentration  calibra-
     tion solutions must be obtained from the Quality  Assurance  Division,
     US EPA Environmental Monitoring Systems Laboratory (EMSL-LV), Las  Vegas,
     Nevada.  However, additional secondary standards  may be obtained from
     commercial sources, and solutions may be prepared in the contractor
     laboratory.   Traceability of standards must be verified against EPA-
     supplied standard solutions.  Such procedures will be documented by
     laboratory SOPs as required in IFB Pre-award Bid  Confirmations, part
     2.f.(4).  It is the responsibility of the laboratory to ascertain  that
     the calibration solutions received are indeed at  the appropriate
     concentrations before they are injected into the  instrument.  Serious
     overloading of the instrument may occur if the concentration  calibra-
     tion solutions intended for a low-resolution MS are injected  into  the
     high-resolution MS.

     7.6.1  The five concentration calibration solutions contain unlabeled
            2,3,7,8-TCDD and labeled 13C,2~1,2,3,4-TCDD at nominal concen-
            trations of 2.5, 5.0, 10.0, 20.0 and 40.0  pg/uL, respectively,
            and labeled 1^C12~2»3,7,8-TCDD at a constant nominal concen-
            tration of 10.0 pg/uL.

                                 D-7

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          7.6.2   Store  the  concentration calibration solutions in 1-mL mini-
                 vials  at 4°C.

     7.7   Column performance  check mixture — this solventless mixture must be
          obtained  fvora the Quality Assurance Division, Environmental Monitoring
          Systems Laboratory, Las Vegas, Nevada, and dissolved by the Contractor
          in 1 mL tridecane.  This solution will then contain the following
          components  (including  TCDDs  (A) eluting closely to 2,3,7,8-TCDD, and
          the first-  (F) and  last-eluting (L) TCDDs when using the columns
          recommended in Section 6.2)  at a concentration of 10 pg/uL of each of
          these  isomers:

          	Analyte	               Approximate Amount Per Ampule

          Unlabeled 2,3,7,8-TCDD                         10 ng

          13C12-2,3,7,8-TCDD                              10 ng

          1,2,3,4-TCDD  (A)                                10 ng

          1,4,7,8-TCDD  (A)                                10 ng

          1,2,3,7-TCDD  (A)                                10 ng

          1,2,3,8-TCDD  (A)                                10 ng

          1,2,7,8-TCDD                                    10 ng

          1,3,6,8-TCDD  (F)                                10 ng

          1,2,8,9-TCDD  (L)                                10 ng


     7.8   Sample fortification  solution — an isooctane solution containing
          the internal  standard  at  a  nominal concentration of 5 pg/uL.

     7.9   Recovery  standard spiking solution — an isooctane solution contain-
          ing the recovery  standard at a nominal concentration of 100 pg/uL.
          Five uL of  this solution will be spiked into the extract before
          HRGC/HRMS analysis.

     7.10 Internal  standard spiking solution — an isooctane solution containing
          the internal  standard  at  a  nominal concentration of 100 pg/uL.  Five
          uL of  this  solution will be added to a fortified field blank extract
          (Section  4.2.1.A.2, Exhibit E).

8.   SYSTEM PERFORMANCE CRITERIA

     System performance criteria are  presented  in two  sections.  One section
     deals with GC  column performance criteria while the other section consists
     of initial calibration criteria.  The  laboratory may use either of the
     recommended columns described  in Section 6.2.   It must be documented that

                                      D-8

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all applicable system performance criteria specified in Sections  8.1,  8.2
and 8.3 have been met before analysis of any  sample  is  performed.   Table 2
provides recommended conditions that can be used to  satisfy the required
criteria.  Table 3 provides a typical 12-hour analysis  sequence.

8.1  GC Column Performance

     8.1.1  Inject 2 uL (Section 6.1.1)  of the column performance  check
            solution (Section 7.7)  and acquire selected ion monitoring
            (SIM) data for m/z 258.930,  319.897, 321.894,  331.937  and
            333.934 within a total  cycle time of 
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     NOTE:  Excessive PFK may cause background noise problems  and contami-
            nation of the source resulting in an increase  in "downtime"
            for source cleaning.

            Using a PFK molecular leak,  tune the instrument to meet  the
            minimum required mass resolution of 10,000 (10Z valley)  at
            m/z 254.986 (or any other mass reasonably close to ra/z 259).
            Calibrate the voltage sweep at least across the mass range m/z
            259 to m/z 334 and verify that m/z 330.979 from PFK (or  any
            other mass close to m/z 334)  is measured within _+5 ppm (i.e.,
            1.7 mmu) using m/z 254.986 as a reference.  Documentation of  the
            mass resolution must then be  accomplished by recording the
            peak profile of the PFK reference peak m/z 318.979 (or any
            other reference peak at a mass close to m/z 320/322). The
            format of the peak profile representation must allow manual
            determination of the resolution, i.e., the horizontal axis
            must be a calibrated mass scale (amu or ppm per division).
            The result of the peak width  measurement (performed at 5
            percent of the maximum) must  appear on the hard copy and
            cannot exceed 31.9 mmu or 100 ppm.

8.3  Initial Calibration

     Initial calibration is required before any samples are analyzed for
     2,3,7,8-TCDD.  Initial calibration  is also required if any routine
     calibration does not meet the required criteria listed in Section 8.6.

     8.3.1  All concentration calibration solutions listed in  Table  1 must
            be utilized for the initial calibration.

     8.3.2  Tune the instrument with PFK  as described in Section 8.2.2.

     8.3.3  Inject 2 uL of the column performance check solution (Section
            7.7) and acquire SIM mass spectral data for m/z 258.930,
            319.897, 321.894, 331.937 and 333.934 using a  total cycle time
            of <_ 1 second (Section 8.3.4.1).  The laboratory must not
            perform any further analysis  until it has been demonstrated
            and documented that the criterion listed in Section 8.1.2 has
            been met.

     8.3*4  Using the same GC (Section 8.1) and MS (Section 8.2) conditions
            that produced acceptable results with the column performance
            check solution, analyze a 2-uL aliquot of each of  the 5  concen-
            tration calibration solutions in triplicate with the following
            MS operating parameters.

            8.3.4.1  Total cycle time for data acquisition must be <_ 1
                     second.  Total cycle time includes the sum of all the
                     dwell times and voltage reset times.

            8.3.4.2  Acquire SIM data for the following selected
                     characteristic ions:
                                 D-10

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                            m/z          Compound

                          258.930        TCDD - COC1

                          319.897        Unlabeled TCDD

                          321.894        Unlabeled TCDD

                          331.937        13C12-2,3,7,8-TCDD,  13C12-1,2,3,4-TCDD

                          333.934        13C12-2,3,7,8-TCDD,  13C12-1,2,3,4-TCDD
                 8.3.4.3  The ratio of integrated ion current  for  m/z  319.897  to
                          m/z 321.894 for 2,3,7,8-TCDD must be between 0.67  and
                          0.90.

                 8.3.4.4  The ratio of integrated ion current  for  m/z  331.937  to
                          m/z 333.934 for 13C1?-2,3,7,8-TCDD and 13C12-1,2,3,4-
                          TCDD must be between 0.67 and 0.90.

                 8.3.4.5  Calculate the relative response factors  for  unlabeled
                          2,3,7,8-TCDD [RRFCI)] relative to 13C12-2,3,7,8-TCDD
                          and for labeled 13C12-2,3,7,8-TCDD [RRF(II)]  relative
                          to 13C2-1,2,3,4-TCDD as  follows:
                                       RRF(I)   -  —
                                                       AIS
                                      RRF(II)
                                                  AIS
where
*  sum of the integrated ion abundances of m/z  319.897  and  m/z  321.894
   for unlabeled 2,3,7,8-TCDD.

 •  sum of the integrated ion abundances of m/z 331.937 and m/z 333.934
             sum o  te  ntegrate
            for 13C12-2,3,7,8-TCDD.
         *  sum of the integrated ion abundances for m/z 331.937 and m/z
            333.934 for 13C12-1,2,3,4-TCDD.

    QIS  -  Quantity of 13C12~2,3, 7,8-TCDD injected (pg).

    QRg  •  quantity of 13C12~1 ,2,3,4-TCDD injected (pg).

     Qx  »  quantity of unlabeled 2,3,7,8-TCDD injected (pg).
                                      D-ll

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    RRF is a dimensionless quantity; the units used to express QJS»  QRS  an<*  Qx
must be the same*

     8.4  Criteria for Acceptable Calibration

          The criteria listed below for acceptable  calibration must  be met
          before analysis of any sample is performed.

          8.4.1  The percent relative standard deviation (RSD) for the response
                 factors from each of the triplicate analyses  for both unlabeled
                 and   C12-2,3,7,8-TCDD must  be less than ^20  percent.

          8.4.2  The variation of the 5 mean  RRFs  for unlabeled 2,3,7,8-TCDD
                 obtained from the triplicate analyses must  be less  than ^20
                 percent RSD.

          8.4.3  SIM traces for 2,3,7,8-TCDD  must  present a  signal-to-noise
                 ratio of >2.5 for m/z 258.930 and  MO for m/z 321.894.

          8.4.4  SIM traces for 13Cj2-2,3,7,8-TCDD  must present a signal-to-
                 noise ratio MO f°* 333.934.

          8.4.5  Isotopic ratios (Sections 8.3.4.3  and 8.3.4.4) must be  within
                 the allowed range.

          NOTE:  If the criteria for acceptable calibration  listed in Sections
                 8.4.1 and 8.4.2 have been met, the RRF can  be considered inde-
                 pendent of the analyte quantity for the calibration concentra-
                 tion range.  The mean RRF from 5 triplicate determinations  for
                 unlabeled 2,3,7,8-TCDD and for 13C12-2,3,7,8-TCDD will  be used
                 for all calculations until routine calibration criteria (Section
                 8.6) are no longer met.  At  such  time, new  mean RRFs will be
                 calculated from a new set of five  triplicate  determinations.

     8.5  Routine Calibrations

          Routine calibrations must be performed at the beginning of a 12-hour
          period after successful mass resolution and GC column performance
          check runs.

          8.5.1  Inject 2 uL of the concentration  calibration  solution which
                 contains 5.0 pg/uL of unlabeled 2,3,7.8-TCDD, 10.0  pg/uL
                 of 13C12-2,3,7,8-TCDD and 5.0 pg/uL 13C12-1,2,3,4-TCDD.
                 Using the same GC/MS/DS conditions as used  in Sections  8.1,
                 8.2 and 8.3, determine and document acceptable calibration  as
                 provided in Section 8.6.

     8.6  Criteria for Acceptable Routine Calibration

          The following criteria must be met  before further  analysis is  per-
          formed.  If these criteria are not  met, corrective action  must be
          taken and the instrument must be recalibrated.

                                      D-12

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          8.6.1  The measured RRF for unlabeled 2,3,7,8-TCDD must  be  within ^20
                 percent of the mean values  established  (Section 8.3.4.6)  by
                 triplicate analyses of concentration calibration  solutions.

          8.6.2  The measured RRF for   C,2~2,3,7,8-TCDD must be within  +20 per-
                 cent of the mean value established by triplicate  analysis
                 of the concentration calibration solutions  (Section  8.3.A.6).

          8.6.3  Isotopic ratios (Sections  8.3.4.3 and 8.3.4.4) must  be  within
                 the allowed range.

          8.6.4  If one of the above criteria is not satisfied, a  second attempt
                 can be made before  repeating the entire initialization  process
                 (Section 8.3).

          NOTE:  An initial calibration must be carried out  whenever  any HRCC
                 solution is replaced.

9.   QUALITY CONTROL PROCEDURES

     See Exhibit E for QA/QC requirements.

10.  SAMPLE PRESERVATION AND HANDLING

     10.1 Chain-of-custody procedures — see Exhibit G.

     10.2 Sample Preservation

          10.2.1 When received, each soil or sediment 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 the temperature is maintained between 25° and  35°C.
                 After a portion of  a sample has been removed for  analysis,
                 return the remainder of the sample to its original container
                 and store as stated above.

          10.2.2 Each aqueous sample will be contained in a  1-liter glass
                 bottle.  The bottles with  the samples are stored  at  4°C in a
                 refrigerator located in a  locked limited-access area.

          10.2.3 To avoid photodecomposition, protect samples from light.

     10.3 Sample Handling

     CAUTION:  Finely divided soils  contaminated with 2,3,7,8-TCDD are hazardous
               because of the potential for  inhalation 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).

          10.3.1 Pre-extraction sample treatment


                                      D-13

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                 10.3.1.1 Homogenization — Although sampling personnel will
                          attempt to collect homogeneous  samples,  the contrac-
                          tor shall examine each sample and judge  if it needs
                          further mixing.

                 NOTE:  Contractor personnel have the responsibility to take a
                        representative sample portion;  this responsibility
                        entails efforts to make the sample as homogeneous as
                        possible.  Stirring is recommended when possible.

                 10.3.1.2 Centrifugation — When a soil or sediment sample
                          contains an obvious liquid phase, it must be
                          centrifuged to separate the liquid from  the solid
                          phase.  Place the entire sample in a suitable centri-
                          fuge bottle and centrifuge for  10 minutes at 2000 rptn.
                          Remove the bottle from the centrifuge.  With a dis-
                          posable pipet, remove the liquid phase and discard
                          it.  Mix the solid phase with a stainless steel
                          spatula and remove a portion to be air-dried, weighed
                          and analyzed.  Return the remaining solid portion to
                          the original sample bottle and  store it  as described
                          in 10.2.1.

                 CAUTION:  The removed liquid may contain TCDD and should be
                           disposed as a liquid waste.

                 10.3.1.3 Weigh between 9.5 and 10.5 g of the air-dried soil
                          sample (+0.5 g) to 3 significant figures.  Dry it to
                          constant weight at 100°C.  Allow the sample to cool
                          in a desiccator.  Weigh the dried soil to 3 signifi-
                          cant figures.  Calculate and report percent moisture
                          on Form B-l.
11.  SAMPLE EXTRACTION
     11.1 Soil Extraction
          11.1.1 Immediately before use,  the Soxhlet apparatus is charged
                 with 200 to 250 mL benzene which is then refluxed for 2  hours.
                 The apparatus is allowed to cool,  disassembled and the benzene
                 removed and retained as  a blank for later analysis if required.

          11.1.2 Accurately weigh to 3 significant figures a 10-g (9.50 g to
                 10.50 g) portion of the  wet soil or sediment sample.   Mix 100
                 uL of the sample fortification solution (Section 7.8) with
                 1.5 mL of acetone (500 pg of   C,2~2,3,7,8-TCDD) and  deposit
                 the entire mixture in small portions on several sites on the
                 surface of the soil or sediment.

          11.1.3 Add 10 g anhydrous sodium sulfate and mix thoroughly  using a
                 stainless steel spoon spatula.
                                      D-14

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     11.1.4 After breaking up any lumps, place the soil-sodium sulfate
            mixture in the Soxhlet apparatus using a  glass  wool plug  (the
            use of an extraction thimble is optional).   Add 200 to 250  tnL
            benzene to the Soxhlet apparatus and reflux for 24 hours.   The
            solvent must cycle completely through the system at least 3
            times per hour.

     11.1.5 Transfer the extract to a Kuderna-Danish  apparatus and
            concentrate to 2 to 3 uL.  Rinse the column and flask  with  5 mL
            benzene and collect the rinsate in the concentrator tube.
            Reduce the volume in the concentrator tube  to 2 to 3 uL.
            Repeat this rinsing and concentrating operation twice  more.
            Remove the concentrator tube from the K-D apparatus and care-
            fully reduce the extract volume to approximately 1 mL  with  a
            stream of nitrogen using a flow rate and  distance such that
            gentle solution surface rippling is observed.

      NOTE:  Glassware used for more than one sample  must be carefully
             cleaned between uses to prevent cross-contamination (Note  on
             page D-10).

11.2  Extraction of Aqueous Samples

      11.2.1  Mark the water meniscus on the side of  the 1-L sample bottle
              for later determination of the exact sample volume.   Pour
              the entire sample (approximately 1 L) into a 2-L separatory
              funnel.

      11.2.2  Mix 100 uL of the sample fortification  solution with 1.5  mL
              of acetone (500 pg of   C\2~2,3,7,8-TCDD) and add the mixture
              to the sample in the separatory funnel.

      NOTE:  A continuous liquid-liquid extractor may be used in place  of
             a separatory funnel.

      11.2.3  Add 60 mL mcthylene 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.
              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-third the volume of the solvent
              layer, the analyst must employ mechanical techniques to
              complete the phase separation.  Collect the methylene
              chloride (3 x 60 mL) directly into a 500 mL Kuderna-Danish
              concentrator (mounted with a 10 mL concentrator tube) by
              passing the sample extracts through a filter funnel packed
              with a glass wool plug and 5 g of anhydrous sodium sulfate.
              After the third extraction, rinse the sodium sulfate with an
              additional 30 mL of methylene chloride  to ensure quantitiative
              transfer.
                                 D-15

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     11.2.4   Attach a Snyder column and concentrate the extract  until
              the apparent volume of the liquid  reaches  1  mL.   Remove the
              K-D apparatus and allow it to drain and cool for at least
              10 minutes.  Remove the Snyder column,  add 50  mL benzene,
              reattach the Snyder column and concentrate to  approximately
              1 mL.  Rinse the flask and the lower joint with  1 to 2 mL
              benzene.  Concentrate the extract  to 1.0 mL under a gentle
              stream of nitrogen.

     11.2.5 Determine the original sample volume by refilling  the sample
            bottle to the mark and transferring  the liquid to  a 1000-mL
            graduated cylinder.  Record the sample volume to the  nearest
            5 mL.

11.3 Cleanup Procedures - Column Cleanup

     11.3.1 Prepare an acidic silica column as follows:   Pack  a 1  cm x 10
            cm chromatographic column with a glass wool  plug,  a layer (1
            cm) of ^804/1^03(1:1), 1.0 g silica gel (Section 7.1.2} and
            4.0 g of 40-percent w/w sulfuric acid-impregnated  silica gel
            (Section 7.1.3).   Pack a second chromatographic  column (1 cm x
            30 cm) with a glass wool plug,  6.0 g acidic  alumina (Section
            7.1.1) and top with a 1-cm layer of  sodium sulfate (Section
            7.4).  Add hexane to the columns until they  are  free  of
            channels and air  bubbles.

     11.3.2 Quantitatively transfer the benzene  extract  (1 mL) from the
            concentrator tube to the top of the  silica gel column. Rinse
            the concentrator  tube with two 0.5-mL portions of  hexane.
            Transfer the rinses to the top of the silica gel column.

     11.3.3 Elute the extract from the silica gel column with  90  mL hexane
            directly into a Kuderna-Danish concentrator.  Concentrate the
            eluate to 0.5 mL, using nitrogen blow-down as necessary.

     11.3.4 Transfer the concentrate (0.5 mL) to the top of  the alumina
            column.  Rinse the K-D assembly with two 0.5-mL  portions of
            hexane and transfer the rinses to the top of the alumina
            columns.  Elute the alumina column with 18 mL hexane  until the
            hexane level is just below the top of the sodium sulfate.
            Discard the eluate.  Columns must not be allowed to reach
            dryness (i.e., a  solvent "head" must be maintained.)

     11.3.5 Place 30 mL of 20-percent (v/v) methylene chloride in hexane
            on top of the alumina and elute the  TCDDs from the column.
            Collect this fraction in a 50-mL Erlenmeyer  flask.

     11.3.6 Certain extracts, even after cleanup by column chromatography,
            contain interferences which preclude determination of TCDD
            at low parts-per-trillion levels. Therefore,  a  cleanup step
            is included using activated carbon which selectively  retains
            planar molecules  such as TCDD.  The  TCDDs are then removed

                                 D-16

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                 from the carbon by elution with toluene.   Proceed  as  follows:
                 Prepare a 18-percent Carbopak C/Celite 545®  mixture by
                 thoroughly mixing 3.6 grams Carbopak C (80/100 mesh)  and  16.4
                 grams Celite 545® in a 40-mL vial.   Activate at  130°C for  6
                 hours.  Store in a desiccator.  Cut off a clean  5-mL  disposable
                 glass pipet at the 4-mL mark.  Insert  a plug of  glass wool
                 (Section 7.3) and push to the 2-mL  mark.   Add 340  mg  of  the
                 activated Carbopak/Celite mixture followed by another glass
                 wool plug.  Using two glass rods, push both  glass  wool plugs
                 simultaneously towards the Carbopak/Celite mixture and gently
                 compress the Carbopak/Celite plug to a length of 2 to 2.5  cm.
                 Preelute the column with 2 mL toluene  followed by  1 mL of
                 75:20:5 methylene chloride/methanol/benzene, 1 mL  of  1:1
                 cyclohexane in methylene chloride,  and 2  mL  hexane.   The  flow
                 rate should be less than 0.5 mL min.~^.  While the column is
                 still wet with hexane, add the entire  eluate (30 mL)  from the
                 alumina column (Section 11.3.5) to  the top of the  column.
                 Rinse the Erlenmeyer flask which contained the extract twice
                 with 1 mL hexane and add the rinsates  to the top of the  column.
                 Elute the column sequentially with  two 1-mL  aliquots  hexane, 1
                 mL of 1:1 cyclohexane in methylene  chloride, and 1 mL of
                 75:20:5 methylene chloride/ methanol/benzene. Turn the  column
                 upside down and elute the TCDD fraction with 6 mL  toluene  into
                 a concentrator tube.  Warm the tube to approximately  60°C and
                 reduce the toluene volume to approximately 1 mL  using a  stream
                 of nitrogen.  Carefully transfer the residue into  a 1-mL
                 mini-vial and again at elevated temperature, reduce the  volume
                 to about 100 uL using a stream of nitrogen.   Rinse the concen-
                 trator tube with 3 washings using 200  uL of  1% toluene in
                 CH2C12*  Add 50 uL tridecane and store the sample  in  a refrig-
                 erator until GC/MS analysis is performed.

12.  ANALYTICAL PROCEDURES

     12.1 Remove the sample extract or blank from storage, allow  it to warm to
          ambient laboratory temperature and add 5 uL recovery standard solution.
          With a stream of dry, purified nitrogen, reduce the extract/blank
          volume to 50 uL.

     12.2 Inject a 2-uL aliquot of the extract into  the GC, operated under the
          conditions previously used (Section 8.1) to produce acceptable  results
          with the performance check solution.

     12.3 Acquire SIM data according to 12.3.1.  Use the same acquisition time
          and MS operating conditions previously used (Section 8.3.4)  to  deter-
          mine the relative response factors.

     12.3.1 Acquire SIM data for the following selected characteristic ions:
                                      D-17

-------
            m/z                        Compound

          258.930                      TCDD - COC1

          319.897                      Unlabeled  TCDD

          321.894                      Unlabeled  TCDD

          331.937                      13C12-2,3,7,8-TCDD,  13C12~1 ,2,3,4-TCDD

          333.934                      13C12-2,3,7,8-TCDD,  13C12-1 ,2,3 ,4-TCDD


     12.4 Identification Criteria

          12.4.1  The retention time (RT)  (at maximum  peak height)  of  the  sample
                 component m/z 319.897 must be within -1 to +3  seconds of the
                 retention time of  the peak for the isotopically labeled  internal
                 standard at m/z 331.937  to attain a  positive  identification of
                 2,3,7,8-TCDD.  Retention times of other tentatively  identified
                 TCDDs must fall within the RT window established  by  analyzing
                 the column performance check solution (Section 8.1).  Retention
                 times are required for all chroma tograms.

          12.4.2  The ion current responses for m/z 258.930,  319.897 and 321.894
                 must reach maximum simultaneously (+_ 1 scan),  and all ion
                 current intensities must be ^ 2.5 times noise  level  for
                 positive identification  of a TCDD.

          12.4.3  The integrated ion current at m/z 319.897  must be between 67
                 and 90 percent of  the ion current response at  m/z 321.894.

          12.4.4  The integrated ion current at m/z 331.937  must be between 67
                 and 90 percent of  the ion current response at  m/z 333.934.

          12.4.5  The integrated ion currents for  m/z  331.937 and 333.934  must
                 reach their maxima within +_ 1 scan.

          12.4.6  The recovery of the internal standard 13C12-2,3,7,8-TCDD must
                 be between 40 and  120 percent.

13.   CALCULATIONS

     13.1 Calculate the concentration of  2,3,7,8-TCDD (or any  other TCDD  isomer)
          using the formula:
                                  cx>
                                           AX
                                       AIg *  W •  RRF(I)
                                      D-18

-------
where:
     Cx  •  unlabeled 2,3,7,8-TCDD (or any other unlabeled TCDD isomer)  concen-
            tration in pg/g for soil/sediment and pg/L for aqueous  samples.

     AX  •=  sum of the integrated ion abundances determined for m/z 319.897
            and 321.894.

     AJS «=  sura of the integrated ion abundances determined for m/z 331.937
            and 333.934 of *3C12-2,3,7 ,8-TCDD (IS - internal standard).

     QIS »  quantity (in picograms)  of 13Cj2-2,3,7 ,8-TCDD added to  the
            sample before extraction (Qxs * 500 pg) .

      W  *  weight (in grams) of dry soil or sediment sample or volume of
            aqueous sample (in liters).
RRF(I)
            calculated mean relative response factor for unlabeled 2,3,7,8TCDD
            relative to   C, 2-2,3,7,8-TCDD.   This represents  the  grand
            mean of the RRF(l)'s obtained in Section 8. 3. A. 5.
     13.2  Calculate the recovery of the internal standard  13C12-2,  3,7,8-TCDD,
           measured in the sample extract,  using the formula:

           Internal standard           A™  * Qr,g
           percent recovery    «  •                     x  100
                                  ARS ' RRF(II) ' QIS

where Ajg and Qjg have the same definitions as above (Section  13.1)

         »  sum of the integrated ion abundances determined for m/z  331.937
            and 333.934 of r3C12-l,2,3,4-TCDD (RS - recovery standard).

    QRg  "  quantity (in picograms)  of   Cj2~l ,2,3,4-TCDD added to the sample
            residue before HRGOHRMS analysis.

    (QRS - 500 pg).

RRF(II)  •  calculated mean relative response factor for labeled  3Cj2~2,3, 7,8-
            TCDD relative to   C12-l ,2,3,4-TCDD.  This represents  the  grand
            mean of the RRF(II)'s calculated in Section 8.3.4.5.

     13.3  If the calculated concentration  of unlabeled 2,3,7,8-TCDD exceeds
           200 pg/g for soils or sediments, or 2000 pg/L for aqueous samples,
           the linear range of response vs. concentration may  have been  exceeded
           and a smaller portion of that sample must be analyzed.   Accurately
           weigh to three significant figures a 1-g portion of the wet soil/
           sediment.  Add the sample fortification solution (Section 11.1.2),
           extract and analyze as discussed for the 10-g sample.  Similarly,
           add the sample fortification solution (Section 11.2.2)  to 100 mL of
           the aqueous sample, extract and  analyze.

                                      D-19

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     13.4  Total TCDD concentration — all positively identified isoraers of TCDD
           must be within the RT window and meet all identification criteria
           listed in Sections 12.4.2, 12.4.3 and 12.4.4.   Use the expression
           in Section 13.1 to calculate the concentrations of the other TCDD
           isomers, with Cx becoming the concentration of any unlabeled TCDD
           isomer.

  Total TCDD  -  Sum of the concentrations of the individual TCDDs.

     13.5  Estimated Detection Limit — For samples in which no unlabeled
           2,3,7,8-TCDD was detected, calculate the estimated minimum detectable
           concentration.  The background area is determined by integrating the
           ion abundances for m/z 319.897 and 321.894 in  the appropriate region
           of the selected ion monitoring trace, multiplying that area by 2.5,
           and relating the product area to an estimated  concentration that
           would produce that product area.

           Use the formula:

                                          (2.5) • (Ax) '  (QIS)
                                         (AIS) ' (RRF(I)) '  (W)

where

     CE  •  estimated concentration of unlabeled 2,3,7,8-TCDD required to
            produce Ax.

     Ax  »  sum of integrated ion abundance for m/z 319.897  and  321.894 in the
            same group of >5 scans used to measure AI§.

     AIS •  sum of integrated ion abundance for the appropriate  ion  character-
            istic of the internal standard, m/z 331.937 and  m/z  333.934.

QlS, RRF(I), and W retain the definitions previously stated  in Section 13.1.
Alternatively, if peak height measurements are used for quantification, measure
the estimated detection limit by the peak height of the noise in the TCDD RT
window.

     13.6  The relative percent difference (RPD) is calculated as follows:

                        |  8j - 82 |            | Si - 82 |
             RPD  -  —_——^————  =  	 x  100
                     Mean Concentration      (Sj + 82)/2

          S} and $2 represent sample and duplicate sample results.

References

1.  "Carcinogens - Working with Carcinogens", Department of  Health,  Education
     and Welfare, Public Health Service, Center for Disease  Control, National
     Institute for Occupational Safety and Health, Publication No. 77-206, Aug.
     1977.
                                     ' D-20

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

3.   "Safety in Academic Che'nistry Laboratories",  American Chemical Society
     Publication, Committee on Chemical  Safety, 3rd  Edition 1979*
                                      D-21

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TABLE 1.  COMPOSITION OF CONCENTRATION CALIBRATION SOLUTIONS


HRCC1
HRCC2
HRCC3
HRCC4
HRCC5
Recovery Standard
13C12-1,2,3,4-TCDD
2.5 pg/uL
5.0 pg/uL
10.0 pg/uL
20.0 pg/uL
40. 0 pg/uL
Analyte
2,3,7,8-TCDD
2.5 pg/uL
5.0 pg/uL
10.0 pg/uL
20.0 pg/uL
40.0 pg/uL
Internal Standard
13C12-2,3,7,8-TCDD
10.0 pg/uL
10.0 pg/uL
10.0 pg/uL
10.0 pg/uL
10.0 pg/uL
               Sample Fortification Solution

              5.0 pg/uL of 13C12-2,3,7,8-TCDD
             Recovery Standard Spiking  Solution

                            '12'
100 pg/uL 13C,2-1,2,3,4-TCDD
            Field Blank Fortification Solutions

          A)  5.0 pg/uL of unlabeled 2,3,7,8-TCDD

          B)  5.0 pg/uL of unlabeled 1,2,3,4-TCDD
             Internal Standard Spiking  Solution

              100 pg/uL of 13C1?-2,3,7,8-TCDD
             (Used only in Section 4.2.1.A.2,  Exhibit E)
                             D-22

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                 TABLE 2.  RECOMMENDED GC OPERATING  CONDITIONS
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°C

70"C

4 min

Rapid increase to 200°C
200°C to 250°C
at 4°C/min

24 min
CP-SIL 88

0.22 urn

50 m x 0.22 mm

28-29 cm/sec
at 240°C

45°C

3 min

Rapid increase to 190°C
190°C to 240°C
at 5°C/min

26 min
                                      D-23

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TABLE 3.  TYPICAL 12-HOUR SEQUENCE FOR 2,3,7,8-TCDD ANALYSIS
1.  Static mass resolution check




2.  Column performance check




3.  HRCC2




4.  Sample 1 through Sample "N"




5.  Column performance check




6.  Static mass resolution check
                                      10/20/84




                                      10/20/84




                                      10/20/84




                                      10/20/84




                                      10/20/84




                                      10/20/84
0700 hrs.




0730 hrs.




0800 hrs.




0830 hrs.




1800 hrs.




1830 hrs.
                            D-24

-------
K>
l/i
                 IM
                  M
                                                  2378
                                                                    1294
                                 «M
                                                     I '
                                                     •M
     CT-SIt 88

      (SO.)
1278
                                                                                                    12(7
                                                                                             •M
          Figure  1.   Selected ion current profile  for n/z 320 and 322 produced by MS analysis for
                      performance check solution using a  50-in CP Si 1-88 fused silica capillary
                      column and conditions  listed  in Table 2.

-------
              IM
7
K>
              10
                                             2371
                                                         — 1234
                                              U7i
                                                                                      CT-2330



                                                                                      (60 •)
                                                                            1271
                  311   IM
                                                      44*
                                                                          4M
      Figure 2.  Selected ion current  profile  for ra/z  320 and 322 produced by MS analysts  of  performance

                 check solution using  a 60-m SP-2330 fused silica capillary column and conditions

                 listed in Table 2.

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                                   APPENDIX B

                          PROPOSED ANALYTICAL PROTOCOL

                            for the Determination of
          2,3,7,8-Tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD)  and  Total
              TCDDs in Soil/Sediment and Water by High-Resolution
              Gas Chromatography/High-Resolution Mass Spectrometry
                                December 1,  1985


     This analytical protocol has been written in the format used in the
Superfund program, as "Exhibit D" of a Statement of Work  which  in turn  is  part
of an Invitation-for-Bid package under the Superfund Contract Laboratory Program.
Also included are other exhibits listed below for the Statement of Work which
have been tailored to meet the specific requirements of this protocol:

EXHIBIT B:  Reporting Requirements and Deliverables
EXHIBIT C:  Sample Rerun Requirements
EXHIBIT D:  Analytical Method
EXHIBIT E:  Quality Assurance/Quality Control Requirements

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     This protocol (Protocol B) is a modification of the protocol  presented  as
Appendix A (Protocol  A).  Examination of the results from the  single-laboratory
evaluation of Protocol A had shown that the minimum amount of  2,3,7,8-TCDD that
could be quantified under the conditions specified in Protocol  A was  5  pg.
However, a requirement existed to lower the quantitation limits to 2  ppt  for
soil and sediment samples and to 20 ppq for aqueous samples.   The  sample  size
should stay at 10 g for soil and sediments and at 1 L for aqueous  samples,
since the effect of larger sample sizes on the extract cleanup efficiencies  is
not known.  Also, the range of the method should overlap with  the  1-ppb lower
limit of the low-resolution analytical  method for TCDD used in the Superfund
Contract Laboratory Program without necessitating second extractions  for  samples
containing higher levels of TCDDs.

     After careful evaluation by EMSL-LV of the requirements and the  options,
the following protocol changes were made:

     o  In Protocol B, the following calibration solutions will be used:

          HRCC1:    2 pg/uL   2,3,7,8-TCDD and 13C12-1,2,3,4-TCDD
                   10 pg/yL   13C12-2,3,7,8-TCDD
          HRCC2:   10 pg/yL   2,3,7,8-TCDD and 13C12-1,2,3,4-TCDD
                   10 pg/yL   13C12-2,3,7,8-TCDD
          HRCC3:   50 pg/yL   2,3,7,8-TCDD and 13C12-1,2,3,4-TCDD
                   10 pg/yL   13C12-2,3,7,8-TCDD

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     HRCC4:  100 pg/yL   2,3,7,8-TCDD and  13C12-1,2,3,4-TCDD
              10 pg/yL   13C12-2,3,7,8-TCDD

o  In Protocol  B, the final  extract volume will  be  10 yL.  The decision
   to select a final  volume  of 10 yL was necessary  in order to comply
   with the above requirements.   It is realized  that such a small volume
   may pose technical difficulties for the analyst.

0  In Protocol  B, the fortification level  of  the internal standard
   13C12-2,3,7,8-TCDD was raised from 500  pg/sample to  1,000 pg/sample.
   This allows analysis of soil  and sediment  samples containing  between
   100 ppt and 1.2 ppb of any TCDD isomer  and of water  samples containing
   between 1 ppt and 12 ppt  of any TCDD isomer by diluting a 2-yL aliquot
   of the remaining extract  concentrate by a  factor of  12 with a solution
   of the recovery standard  (100 pg/yL of  13C12-1,2,3,4-TCDD in  tridecane),
   Recoveries will  be reported using the data generated from the first
   injection.  Thus,  the decision to dilute an aliquot  of the 10-yL final
   extract will  not be based on  the concentration of 2,3,7,8-TCDD or total
   TCDD in the sample, but on the concentration  of  the  most abundant TCDD
   isomer in the 10-yL final extract volume.   This  will  eliminate un-
   necessary dilutions of the sample extract  and analyses for samples
   containing between 100 ppt and 250 ppt  for soil  and  sediment  and 1 ppt
   and 2.5 ppt for water samples of a TCDD isomer,  but  for which the
   recoveries were low.

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               EXHIBIT B
Reporting Requirements and Deliverables

-------
1.   SCOPE AND APPLICATION

     The Contractor shall provide reports  and  other  deliverables  as  specified
     in the Contract Reporting  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 selected ion
     current profiles (also  called selected  ion  monitoring  traces),  shall be
     clearly labeled with the  Sample Management  Office Case number and associated
     Sample/Traffic Report number(s).  If  documentation is  submitted without
     the required  identification, as specified above, resubmission shall be
     required.

     The Contract  Reporting  Schedule (Section  2)  specifies  the  numbers of
     copies required, the delivery schedule  and  the  distribution  of  all required
     deliverables.

     1.1  Sample data package  — Hard copy analytical data  and  documentation
          are required as described below.

     NOTE:  This analytical  protocol is designed for the  receipt  and analysis
            of samples by batches.  Therefore, it is desired  that sample data
            from samples in  the same batch be  reported together,  i.e., on the
            same reporting form.  However, contract  accounting  and billing are
            based  on the sample unit.

          1.1.1  Case narrative:  Contains the Case  number, Dioxin Shipment
                 Record numbers, Contract  number and detailed documentation of
                 any quality control, sample,  shipment and/or analytical pro-
                blems 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.

          1.1.2  Results of  initial triplicate analyses of  four (4)  concentration
                 calibration solutions (Form H-2), routine  calibration solutions,
                 (Form H-3), including all selected  ion current profiles or
                 selected ion  monitoring (SIM) traces, calculated relative
                 response factors (RRF),  and computer-generated quantification
                 reports (or manual calculations).

          1.1.3  Completed data reporting  sheets (Forms H-l,  H-4, and H-5, H-8
                 and H-9) with  appropriate SIM traces (including  the lock mass
                 SIM traces).   Data results for levels less than  10  ppt but
                above the quantitation limit  (Section 1.1, Exhibit  D) attained
                 for that sample shall be  reported to two (2) significant
                 figures; results greater  than 10 ppt 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 SIM trace
                 shall include  computer-generated header  information indicating
                 instrumental  (GC and MS)  operating  parameters  during data

                                      B-l

-------
            acquisition.  When samples are analyzed more than once,  all
            sample data shall be reported.  Rejected sample runs must  be
            separated and attached to the back of the data package and
            marked on the SIM trace as "Rejected," with an explanation of
            the reasons for the rejection.

     1.1.4  SIM traces generated during each GC column performance check
            analysis; peak profile outputs of the reference signal used
            to document the nass resolution.

     1.1.5  Documentation of acceptable MS calibration (Section 8, Exhibit
            D, and Exhibit E) for each confirmatory analysis.  As
            applicable, submit peak matching box settings  and calculations
            for accurate mass assignments and any other related printouts.
            State, in ppm, the level of mass accuracy achieved (Section
            8.2.2, Exhibit D).

     1.1.6  A chronological list of all analyses performed (Form H-6).  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, performance check
            solution, or other pertinent analytical data.   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 GC column performance check solution  ana-
            lyses, one preceding and one following the sample/blank  ana-
            lyses.  If multiple shifts are used, the ending GC column
            performance check sample analysis from one 12-hour period
            shall serve as the beginning analysis for the  next 12-hour
            period; see Exhibit D, Section 8, for system performance
            criteria.  The same schedule applies to the mass resolution
            check analysis.  See Section 8.2.2, Exhibit D.

     1.1.7  Verification of recovery of TCDDs from cleanup columns
            (Section 11.3, Exhibit D, and Section 4.2.1.2.2, Exhibit E).

1.2  Sample extracts and unused sample portions — Unused  portions of
     samples and sample extracts shall be retained by the  Contractor for
     a period of six months after receipt.  When directed  in writing by
     the Project Officer (PO) or Sample Management Office  (SMO), the
     Contractor shall ship (not at Contractor's expense but in accordance
     with Department of Transportation Regulations) specific samples
     and/or extracts to specified locations and persons.  After six  months,
     upon obtaining PO or SMO clearance, remaining samples and extracts
     shall be disposed of by the Contractor at Contractor's expense, in
     accordance with applicable regulations concerning the disposal  of
     such materials.

1.3  Document Control and Chain-of-Custody Package — The  Document Control
     and Chain-of-Custody Package includes all laboratory  records received
                                 B-2

-------
          or generated for a specific case,  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, corre-
          spondence, and the document inventory  (Exhibit G).

     NOTE:  Pages from logbooks or bench sheets  kept  exclusively in a  high-
            hazard area (containment facility) need not be copied.

     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, and samples which  required special  cleanup.
          All samples shall be identified by the appropriate  EPA sample, case
          and batch/shipment numbers.

     1.5  Daily Sample Status Report — In response to a 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 the
          daily sample status information in a written form that same  day using
          first-class mail service.  The required Daily Sample Status  informa-
          tion 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.

2.   In accordance with applicable delivery  requirements, the Contractor shall
     deliver specified items per the following Contract Reporting  Schedule
     (Section 2.1).  Recipients include the  CLP  Sample Management  Office, the
     EMSL/LV QA Division, the appropriate Regional Technical Officer and NEIC.

     2.1  Contract Reporting Schedule

                          CONTRACT REPORTING SCHEDULE

Item                             Delivery                Report Distribution
 No.  Report     No. Copies	Schedule	SMO   EMSL/LV  Region  NEIC

 1  Sample Data     3     30 days after validated     XXX
     Package                sample receipt date
                                    -OR-
                            10 days after initial     XXX
                               data due date

2   Sample Extracts        Within 180 days after            As directed
                           analysis, 7 days  after
                          request by Project Officer
                                  or SMO

                                                                  (Continued)

                                      B-3

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                    CONTRACT REPORTING SCHEDULE (Continued)
Item
No.
Report
No.
Copies
Delivery
Schedule
Report Distribution
SMO
EMSL/LV
Region
NEIC
    Document       1
     Control &    Pkg
     Chain-of-
     Custody Package

    Monthly        2
     Sample Status
     Report
   7 days after request by
       Project Officer
          or SMO
   5 days following end of
     each calendar month
    Daily Sample
     Status Report
  Verbal and/or written
upon request by SMO or PO;
maximum frequency is daily.
As directed
NOTE:  All results shall be reported total and complete.
     2.2  Addresses for distribution
          SMO
                   EMSL-LV
        NEIC
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
          US EPA EMSL-LV QA Division
          Box 15027
          Las Vegas, NV 89114
           Attn: Data Audit Staff
   US EPA NEIC
   Bldg. 53
   Box 25227
   Denver Fed. Center
   Denver, CO 80225
          For overnight deliveries,  use
           street address:
          944 E. Harmon Ave.
          Executive Center
          Las Vegas, NV 89109
          Regional Technical Officer — Following contract award and prior to
          Contractor's receipt of the first batch of samples,  the Sample Manage-
          ment Office will provide the Contractor with the list of Technical
          Officers for the ten EPA Regions.  SMO will provide  the Contractor
          with updated Regional address/name lists as necessary throughout the
          period of the contract.

3.   FORM INSTRUCTION GUIDE

     This section includes specific instructions for the completion of all
     required forms.  These include instructions on header information as
     well as specific details to the bodies of individual forms.  Instructions
     are arranged in the following order:
                                      B-4

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Data Summary (Form H-l)
Initial Calibration Summary (Form H-2;  2 pages)
Routine Calibration Summary (Form H-3)
GC and Mass Resolution Check Summary (Form H-A)
Quality Control Summary (Form H-5)
Chronological List of All Analyses Performed (Form H-6)
GC Operating Conditions (Form H-7)
HRMS TCDD Calibration Report Form (Form H-8)
High-Resolution MS TCDD Data Report Form (Form H-9)

3.1  Data Summary (Form H-l)— This form is used  for summarizing  the
     results from all samples in the batch.  The  detailed  results are
     available on Form H-8 for each sample.

     Complete the header information at the top  of the page,  including
     laboratory name, case number and batch/shipment number  (from the
     dioxln shipment record), and matrix (soil,  sediment,  water).

     Complete the form using one horizontal row  for each sample.

     The SMO sample number should be suffixed with the appropriate letter
     code as needed.

     The TCDD retention time should be  reported  in minutes and  seconds.

     TCDD levels are reported as parts  per trillion (ppt)  regardless of
     the matrix.  Total TCDD concentration (in ppt) is the sum  of the
     concentrations of all TCDDs reported on Form H-9.

     The S/N criteria apply to m/z 259, 320, 322  (for unlabeled TCDD)
     and m/z 322 and 334 (internal and  recovery  standards).   The  symbols
     used are:  (+) all S/N ratios are  2.5 or greater including all TCDDs
     present, (-) S/N ratio for native  2,3,7,8-TCDD, the internal or the
     recovery standard are less than 2.5, (0) other suspected TCDDs are
     present but did not meet the S/N criteria.

     The file name is the HRGC/HRMS file name and is used  for tracking
     results and raw data.

     The comments column should be used for any  remarks  specific  to a
     particular sample.

3.2  Initial Calibration Summary (Form H-2):  Page 1 —  The  header infor-
     mation should be filled in.  The column headings are  similar to those
     on Form H-l.

                Ax ' QlS
     RRF(I)  =  	
                QX ' AIS
                                 B-5

-------
                 Aj s * QRg
     RRF(II)  =  	       (Section 8.3.4.5,  Exhibit D)
                 QlS * ARS

     Page 2 — The header information should be filled in.  For each RRF,
     the mean, percent relative standard deviation  (%RSD)  and number of
     runs (N) are reported; N must be at least three (3)  for each HRCC
     solution.  The grand means (RRFs) are the mean of the individual
     means and are reported with their %RSD and N.   The routine calibra-
     tion relative response factor permissible ranges are  also reported
     (Section 8.3.4.8, Exhibit D).

3.3  Routine Calibration Summary (Form H-3) — The  header  information
     Includes case and batch numbers in addition to the laboratory and
     instrument identification.

     The columns are the same as on Page 1 of Form  H-2. The results
     reported are for the routine calibration runs  rather  than the initial
     calibration.  The calculated RRF(I) and RRF(II) must  be within the
     routine calibration relative response factor permissible ranges
     (Section 8.3.4.8, Exhibit D) and other criteria listed in Section
     8.6, Exhibit D must be met before further analysis is performed.

3.4  GC and Mass Resolution Check Summary (Form H-4) — The header informa-
     tion should be filled in.  The TCDD isomer resolution (% valley) is
     measured from the column performance check solution  (Section 8.1.2,
     Exhibit D).  The resolving power and mass measurement error are measured
     using PFK (or equivalent) (Section 8.2, Exhibit D).

3.5  Quality Control Summary (Form H-5) — The items should be completed
     as indicated.  The "other interferences" should be included even if
     they only occur at one mass.

     Form H-5 in conjunction with Form H-9 is used  to report results
     relative to the fortified field blank pair and rinsate analyses.

     The total TCDD retention time window is a window that includes all of
     the TCDD isomers and is based on the first and last eluting isomers
     in the GC column performance check solution using the conditions sum-
     marized in Form H-7.  All materials used should be recorded in the
     standard/reagent QC table.  Standards provided by EPA should be
     listed, however, the QC columns may be left blank as  these are refer-
     ence materials.

3.6  Chronological List of All Analyses Performed (Form H-6) — The
     header information should be filled in.  If more than one instrument
     is used, use one form per instrument.

     The "Analysis Identification" column should contain enough information
     for the data user to clearly identify the analysis,  I.e., HRCC 2
     Routine Calibration, Fortified Field Blank A,  Fortified Field Blank B,
                                 B-6

-------
          Reanalysis of Sanple //I, 2, 3, A, etc.   The "SMO //" column should be
          used only for samples etc. which have an assigned SMO sample number.

     3.7  GC Operating Conditions (Form H-7) — This form must be filled out to
          describe the GC operating conditions used to analyze a batch of
          samples and to analyze the GC performance evaluation check solution.

     3.8  HRMS TCDD Calibration Report (Form H-8) — This form is to be filled
          in for each initial and routine calibration analysis made.  It will be
          the first page of the chromatograms and calculations for that analysis.
          It is suggested that this form be used as a worksheet for completing
          Forms H-2 and H-3.  S/N ratios greater than five (5) may be reported
          with a (+); S/N ratios of five or less must have a numerical value
          reported with accompanying chromatograms scaled so that the measure-
          ments may be checked by the data user.

     3.9  High-Resolution MS TCDD Data Report (Form H-9) — This form contains
          the details of the data reported in sunmary on Form H-l.  It will be
          the first page of the chromatograms and calculations for each sample
          including the fortified field blank pair samples.  All data presented
          (retention times, areas, and S/N ratios) must also be available on
          the accompanying chromatograms.  The chromatograms must be scaled
          so that the data user may check any S/N ratios that are near or below
          five (5).  It is suggested that this form be used as a worksheet for
          completing Form H-l.

A.   REPORTING REQUIREMENTS SUMMARY:

     Items that must be included with the data package:

     4.1  Complete identification of the samples analyzed (sample numbers and
          type).

     A.2  The dates and times at which all analyses were accomplished.  This
          information should also appear on each selected ion current profile
          Included with the report.

     A.3  Raw mass chromatographic data which consist of the absolute peak
          heights or peak areas of the signals observed for the ion masses
          monitored.

     A.A  The calculated ratios of the intensities of the M+0 to (M+2)+0
          molecular ions for all TCDD isomers detected.

     A.5  The calculated concentrations of native 2,3,7,8-TCDD and other TCDD
          isomers for each sample analyzed, expressed in picograms TCDD per gram
          of sample (that is, parts per trillion), as determined from the raw
          data.  If no TCDDs are detected, the notation "Not Detected" or
          "N.D." is used, and the minimum detectable concentrations (or detection
          limits) are reported.
                                      B-7

-------
                                                             HIGH  RESOLUTION
                                                         FORM  H-l  DATA SUMMARY
                                                       HRGC/HRMS  DIOXIN  ANALYSIS
         Lab:
 Cawff
         B0tch/Shtpni8ftt w.
                            Matrix:
SMO
Number















TCOO
2.3.7.8 (IS)






























PPt
2.3.7.B-TCDD
Meaa. DL






























Relative km
pot Abundance Ratio*
Total 320 332 332 %Rec. S/N m*t. Anaryei* Re
TCDO 322 334(18) 334(RS| (IS) Criteria ID Date Time Name Comment*





































































































































































B>
I
00
           RB-  Reagent Blank
            N-  UnUbetod TCDO Spfta
            D -  Duplicata
           FB-  Raid Blank
           8R •  Sampto Rerun
           ER -
NO- NotDatscted
 DL- DatactionLimh
MB - Mattwd Blank
Rac- Recovery
Matrix:  8- Soil
       W- Water
       O- Other
S/N Criteria: report (») rl all S/N > 2.S
           report (-) if 2.3.7.8 TCDD.
                   "C,J-2.3.7.8-TCOO or
                   "C,,-1.2.3.4-TCODS/N < 2.S
           report (0) if other TCDO* arc •uapectad
                   not to meet criteria

-------
                                                            HIGH RESOLUTION
                                               FORM  H-2  INITIAL  CALIBRATION  SUMMARY
                                                                                                                            Iof2
           Lab:
Contract ff:
Instrument ID:
w
VO
Catibratfon Rte m/s m/t m/t S/N
Standard Name Date Tlnw 320/322* 332/334(18)* 332/334(RS»* Criteria RRF|l)b RRF(II)C Comments






































































































































































































            * km ratio* mint be hi the rang* of 0.67 to 0.90
            b 2.3.7.8-TCOD ww»u« '»C1Z-2.3.7.B TCDD
            c "C12-2.3.7.8 TCDD vvrau* "C12-1.2.3.4 TCDD
        S/N Criteria: report f+) if greater than 2.6
                  report HH torn than 2.5

-------
                                                           HIGH RESOLUTION
                                               FORM  H-2  INITIAL  CALIBRATION  SUMMARY
                                                                                                         page2of2
                        Lab:
                        Date of Intitial Calibration:
                      Contract #:
           Instrument #:
                                        RRF (I) Mean
                      %RSD
      RRF (II) Mean
                                                                                              %RSO
                          HRCC1

                          HRCC2

                          HRCC3

                          HRCC4
w
I
RRF (I) Grand Mean:

%RSD: 	

N: 	
RRF (II) Grand Mean:

*RSD: 	

N: 	
                        Rotitrn* CcNbratkm P»rm)Mib>» Rwtge:
                            RRF (I) = 2.3.7.8-TCDD w«
                            "C,,-2.3.7.8-TCDD
                                         Routine Calibration PonrriMible Range:
                                             RRF (II) = «»C -2.3.7.8-TCDD w»

-------
                                                         HIGH  RESOLUTION

                                             FORM  H-3 ROUTINE CALIBRATION SUMMARY
          Lab
w
 i

Calibration Ffla m/z m/i m/i S/N
Standard Nama Oat* Tfcna 320/322* 332/334(15)* 332/334(RS|* Criteria RRF(l)b RRFfllF Comnwnta






































































































































































































• l«« r_Mna m.n* ha In tha >mnn_ «« A AT tn H On •! /M OriMri. nnnrt III H 111.. 1.1 than ? K
           • 2.3.7.8-TCOO VWMM "€,,-2.3.7.8-1000

           e "Ct2-2.3.7.8-TCDO vw*tn "C12-1.2.3.4-TCDD
report (•) tf ton thfln 2.5

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                           HIGH RESOLUTION
            FORM H-4 GC AND MASS RESOLUTION CHECK SUMMARY
Lab

TCDD Isomer Revolving 'Man
Intt. Sol. File Revolution Power Measurement
Oat* ID IO Time Name (% Valley) at 10% Valley Error (PPM)
















































































































































































































Mttf Utttf *?» "«•»» iH^murmmM* MMT ealeuliiian
                                B-12

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                                       HIGH RESOLUTION
                            FORM  H-5 QUALITY  CONTROL SUMMARY
Lab:
                                  Case*
Batch #
Number of samples in batch: _
Mean S of recovery for the I.S.:
                                                          * of data points:
Fortified field blank A. K recovery ("C,2-2.3.7.8-TCDD):

        Contamination by 1,2.3.4-TCDD
                  '»C12-1.2.3.4-TCDD
                                                                     SMO Sample ft:
                                                Yes
                                                                                   Estimated
                                                                                   Concentration (ppt)
         Other interferences
                          Retention times:
                          Estimated concentrations (ppt):

Fortified field blank B, % recovery ("C.,-1.2.3.4-TCDD|: _
                                                                . SMO Sample ft:
        Contamination by 2.3.7.8 TCDD
                  "C12-2.3,7.8-TCDD
                                         [_J    []]
               Estimated
               Concentration (ppt)
         Other interferences
                          Retention times:  	
                          Estimated concentration* (ppt):
Rinsate, S recovery:
                                                 SMO sample ft:
         Contamination by 2.3.7.8-TCDD
                         Other TCDD
                                                                                   Concentre lion
                                                                                   (pg/mL)
Duplicate analysis,  SMO sample ft:
                                               "C12-2,3,7.8-TCDD Mean Recovery.
                        Percent Relative difference "C12-2.3.7.8-TCDD (Recovery)	
                        Percent relative difference iaC12-2.3.7.8-TCDD (Concentration)
                        Percent relative difference Total TCDD (Concentration)	
Method blank file name

HRMS Lab.
Standard /Reagent Number or Origin Date of OC File Results of
Type Mfg. ft OC Name QC






















































                                                                                     Continue as needed
                                               B-13

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                             HIGH RESOLUTION
        FORM H-6  CHRONOLOGICAL  LIST OF  ALL ANALYSES PERFORMED
Lab:
Instrument ID:
   Case*
        Batch #
      File
     Name
  Analysis
Identification
 SMO
Number
Date
                                                                          Time
                                    B-14

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                             HIGH  RESOLUTION
                    FORM H-7 GC OPERATING CONDITIONS
Lab:	 Instrument ID:
GC Column:	
Film Thickness:
Column Dimensions.
Initial Column Temperature:
Temperature Program:  	
Injector Temperature:  	
Interface Temperature:  	
Injection Mode: 	
Injection Volume:
Splittess Valve Closed Time:
Septum Purge Flow:  	
Injector Sweep Flow:	
Carrier Gas Flow Rate (ml/min or cm/sec):
                                  B-15

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                            HIGH  RESOLUTION
            FORM H-8  HRMS TCDD CALIBRATION  REPORT FORM
Lab:
Case*:
Batch/Shipment #:
Instrument ID:  	
    Calibration:
       Initial
       Routine

2.3.7.8-TCDD
   m/z 258.930
       319.897
       321.894
»C12-2.3.7.8-TCDD
   m/i331.937
       333.934
«C12-1.2.3.4-TCDD
   m/z 331.937
   m/z 333.934
                Calibration Solution:
               	 GC Column: —
        	 Date of Initial Calibration: .
         Analysis Date:	 Time:
                       File Name
Retention
  Time
Area
Ratios
                                320
                                322
                                334
                                332
                                334
(•) H S/N i* graatar than 8. amar (+); H la** than S. »nt»r tha tnaaaurvd ratio
                                   B-16

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                                     HIGH RESOLUTION
              FORM  H-9  HIGH  RESOLUTION MS TCDD DATA  REPORT  FORM
Lab:  	
Ca*e#:	
Batch/Shipmentft:
        I ID: 	
                                            SMO Sample ft:
                                  Matrix:
                                 Soil
Aliquot
 Circle
  One
g     L
                                        Percent
                                                                       Time:
                                                                  (xtrmrtinn D»Wi
MMtured ppt 2.3.7.8-TCDD:
E«tim»ted Toul TCDO (ppt):
2.3.7.8-TCDD
     m/i 258.930
         319.897
         321.894
                                                Detection Limit 2.3.7.8-TCDD:
                                               	 Report Date:	
                        Retention
                          Time
                                              Area
                                                                 Ratios
                                                        S/N'
'«C12-2.3.7.B-TCDD
     m/i331.937
         333.934
                                                            320.
                                                            322
                                                            332
                                                            334
'»C,j-1.2.3.4-TCDD
     m/t 331 .937
     m/i 333.934
Percent Recovery "C12-2.3.7.8-TCDD:
                                           Other TCDDs
Retention 320 S/N* 269 S/N* 320 S/N* 322 Concentration
Time 322 (ppt)












































































































*H S/N is greater than S. enter |»); H lew than S enter the measured ratio
                                             B-17

-------
4.6  The same raw and calculated data which are provided for the actual
     samples will also be reported for the duplicate analyses, the method
     blank analyses, the fortified field blank pair and rinsate analyses,
     and any other QA or performance sample analyzed in conjunction with
     the actual sample set(s).

4.7  The recoveries of the internal standard (  C12~2»3»7»8~TCDD) in percent.

4.8  The calibration data, including relative response factors calculated from
     the calibration procedure described in Section 8.3, Exhibit D.  Data
     showing that these factors have been verified at least once during each
     12-hour period of operation must be included (Section 8.5, Exhibit D).
     Exact mass measurement error.  Include peak matching box settings
     and calculations as appropriate.

4.9  The calculated dry weight of the original soil or sediment sample portion
     based on the dry weight determination of another sample portion of approxi-
     mately equal wet weight.  The exact volumes of the water and rinsate
     samples analyzed.

4.10 Documentation of the source of all TCDD standards used and available
     specifications on purity.

4.11 In addition, each report of analyses will include the following selcted
     ion current profiles:  1) those obtained from all samples analyzed, 2)
     those from each GC column performance check, and 3) those from the
     calibration solutions.  The peak profile from each mass resolution
     check must also be part of the data package.

4.12 Identify which HRGC/HRMS system was used for the analyses (manufacturer
     and laboratory identification number of system - 01, 02, 03, etc.).

4.13 GC operating conditions such as type of GC column, film thickness, column
     dimensions, initial column temperature, temperature program, injector
     temperature, interface temperature, injection mode and volume, valve time
     (valve flush), septum purge flow, flow rate, and total injector flow
     should be provided (Form H-7).
                                 B-18

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        EXHIBIT C
Sample Rerun Requirements

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1.   SCOPE AND APPLICATION

     The Contractor shall be required to reextract and/or perform additional
     cleanup and reanalyze certain samples 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 "sample rerun"  shall indicate
     sample extraction of a fresh 10-g soil or sediment  portion or 1-L aqueous
     sample, followed by cleanup and analysis, and the term "extract reanalysis"
     shall indicate analysis of another aliquot of the final  extract.

     In situations where the sample rerun is required due to  matrix effects,
     interferences or other problems encountered because of very complex samples,
     the Government will pay the Contractor for the sample reruns.  Such sample
     reruns shall be billable and accountable under the  specified contract
     allotment of automatic reruns.

     In situations where the sample rerun or the extract reanalysis Is required
     due to Contractor materials, equipment or instrumentation problems, or
     lack of contractor's adherence to specified contract procedures, the
     sample rerun or extract reanalysis shall not be billable under the terms
     of the contract.

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

2.   Required Sample Reruns and Extract Reanalyses

     2.1  Automatic sample reruns and extract reanalyses that may be billable
          as such under the contract.

          2.1.1  If the percent recovery for the internal standard   C12~2, 3,7,8-
                 TCDD was outside of the acceptance limits of >bQ percent  and
                 £120 percent, the Contractor shall reextract ITnd reanalyze the
                 sample.  If the percent recovery for the sample rerun is  still
                 outisde the acceptance limits, then both analyses can be  billed
                 if the recoveries from both analyses are either <40% or >120%.
                 If, however, the percent recovery for the sample rerun is
                 within the acceptance limits, or if it is still outside the
                 acceptance limits but the percent recoveries from the original
                 analysis and the sample rerun are not both either <40% or
                 >120%, then the sample rerun may not be billed.

          2.1.2  If the internal standard was not found upon monitoring m/z
                 331.937 and 333.934, the Contractor shall reextract and
                 reanalyze the sample.  If the internal standard is not
                 found in the sample rerun, the sample rerun is billable.   If
                 the internal standard is found in the sample rerun, then  the
                 sample rerun is not billable.
                                      C-l

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     2.1.3  If either one of the isotope abundance ratios for m/z 319.897/
            321.894 or for 331.937/333.934 is less than 0.67 or greater
            than 0.90 and all other criteria contained in Section 12.4  of
            Exhibit D are met, then the extract shall be reanalyzed.   If
            both ion abundance ratios now meet the criterion, these values
            shall be reported as the isotope abundance ratios, and the
            Contractor shall not bill the Government for the extract
            reanalysis.  If the ratio in question is still outside the
            criterion, the Contractor shall rerun the sample (Section 7.2,
            Exhibit E).  If either one of the ratios determined from the
            sample rerun is still outside the acceptance limits,  then
            both runs and the extract reanalysis can be billed if the
            corresponding isotope abundance ratios from both runs are
            either <0.67 or >0.90.  If, however, both isotope abundance
            ratios from the sample rerun meet the criteria, or if both
            corresponding isotope abundance ratios from the original  run
            and the sample rerun are not both either <0.67 or >0.90,
            then the extract reanalysis and the sample rerun may not  be
            billed.

     2.1.4  If the recoveries of 2,3,7,8-TCDD (Section 4.2.1.1.3.1,
            Exhibit E) and/or 1,2,3,4-TCDD (Section 4.2.1.2, Exhibit  E)
            in the fortified field blank pair are <40% or >120%,  the
            Contractor shall reextract and reanalyze a second portion of
            the field blank sample (Section 4.2, Exhibit E).  If  the
            percent recoveries for the sample rerun are still outside the
            acceptance limits, then both analyses can be billed as long
            as the recoveries from both analyses are either <40% or >120%.
            If, however, the percent recoveries for the sample rerun
            are within the acceptance limits, or if they are still outside
            the acceptance Units but the percent recoveries from the
            original run and the sample rerun are not both either <40%
            or >120%, then the sample rerun may not be billed.

            NOTE:  Fortified field blanks as described in Sections
                   4.2.1.1.4 and 4.2.1.2.2, Exhibit E, can never be billed.

2.2  Automatic sample extract dilution and HRGC/HRMS analysis, billable as
     such under the Contract.

     If any individual or group of coeluting TCDD isomer concentrations in
     the 10-uL final extract exceeds 100 pg/uL, the analyst will perform a
     dilution as specified in Section 13.3, Exhibit D, and reanalyze  the
     diluted portion using HRGC/HRMS.


2.3  Sample reruns and/or extract reanalyses to be performed at Contractor's
     expense (i.e., not billable under the terns of the contract).

     2.3.1  If the method blank contains any signal in the TCDD retention
            time window at or above the method quantitation limit (2  ppt
                                 C-2

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       for soil and sediment and 20 ppq  for aqueous  samples),  the
       Contractor shall rerun all positive  samples in  the  batch  of
       samples (Section 4.1.2, Exhibit E).

2.3.2  If the system performance using the  GC column performance
       check (PC) solution does not meet specified criteria,  the
       Contractor shall take corrective  action,  demonstrate acceptable
       GC column performance, and reanalyze the  extracts from all
       positive samples run during the time period between the last
       acceptable PC run and the unacceptable PC run (Section 2.4,
       Exhibit E).

2.3.3  If a false positive is reported for  an uncontaminated  soil
       (blind QC) sample,  upon notification by the Sample  Management
       Office the Contractor shall reextract and reanalyze all samples
       reported as positive in the associated batch  of samples
       (Section 8.1.1, Exhibit E).

2.3.A  If the analysis results for a performance evaluation blind  QC
       sample fall outside of EPA-established acceptance windows,  upon
       notification of the Sample Management Office  the Contractor
       shall reextract and reanalyze the entire  associated batch
       of samples (Section 8.4.1, Exhibit E).

2.3.5  If the isotope abundance ratio for m/z 319.897/321.894 or for
       331.937/333.934 is  less than 0.67 or greater  than 0.90, and
       all other criteria  contained in Section 12.4  of Exhibit D are
       met, then the extract shall be reanalyzed. If  the  ion abundance
       ratio in question now meets the criterion, this value  shall  be
       reported as the isotope abundance ratio,  and  the Contractor
       shall not bill the  Government for the extract reanalysis.

2.3.6  If the system performance mass resolution check does not  meet
       the specified criterion, the Contractor shall take  corrective
       action, demonstrate acceptable mass  resolution  and  reanalyze
       the extract from all positive samples analyzed  during  the time
       period between the  last acceptable pass resolution  check  and
       the unacceptable mass resolution  check (Section 2.4,
       Exhibit E).
                            C-3

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                         EXHIBIT D
                     Analytical Method

2,3,7,8-Tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD)  and Total
  TCDDs in Soil/Sediment and Water by High-Resolution Gas
      Chromatography/High-Resolution Mass Spectrometry

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                                   EXHIBIT D



Section                       Subject                              Page


   1               Scope and Application	     D-l

   2               Summary of Method	     D-l
                                                                    *
   3               Definitions	     D-2

   A               Interferences. ..........  	     D-3

   5               Safety	     D-3

   6               Apparatus and Equipment.	     D-4

   7               Reagents and Standard Solutions	     D-6

   8               System Performance Criteria	     D-9

   9               Quality Control Procedures 	     D-14

  10               Sample Preservation and Handling 	     D-14

  11               Sample Extraction	     D-15

  12               Analytical Procedures	     D-18

  13               Calculations	     D-19

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1.   SCOPE AND APPLICATION

     1.1  This method provides procedures for the detection  and  quantitative
          measurement of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD; CAS
          Registry Number 1746-01-6;  Storet  number 3475)  at  concentrations  of
          2 pg/g (2 parts per trillion) to 100 pg/g (100  parts per  trillion)
          in 10-g portions of soil and sediment and at  20 pg/L (20  parts  per
          quadrillion) to 1000 pg/L (1 part  per trillion) in 1-L samples  of
          water.  Dilution of an aliquot of  the final extract permits  measure-
          ment of concentrations up to 1.2 ng/g (1.2 parts per billion) or  12
          ng/L (12 parts per trillion), respectively.   This  method  also allows
          the estimation of quantities of total TCDD present in  the sample.
          Samples containing concentrations  of any individual TCDD  isomer or
          group of coeluting TCDD isomers greater than  1.2 ng/g  or  12  ng/L  must
          be analyzed by a protocol designed for such concentration levels,
          with an appropriate instrument calibration range.

     1.2  The minimum measurable concentration is estimated  to be 2 pg/g  (2
          parts per trillion) for soil and sediment samples  and  20  pg/L (20
          parts per quadrillion) for water samples, but this depends on kinds
          and concentrations of interfering compounds in  the sample matrix.

     1.3  This method is designed for use by analysts who are experienced in
          the use of high-resolution gas chromatography/high-resolution mass
          spectrometry.

     CAUTION:  TCDDs are assumed to be extremely hazardous.   It  is  the labora-
               tory's responsibility to ensure that safe  handling procedures are
               employed.

2.   SUMMARY OF METHOD

     One thousand pg of   C,2~2,3,7,8-TCDD (internal standard) are  added  to a
     10-g portion of a soil/sediment  sample (weighed to 3 significant  figures)
     or a 1-L aqueous sample, and the sample is extracted with 200  to 250 mL
     benzene using a Soxhlet apparatus for soils and sediments with a minimum
     of 3 cycles per hour, or with methylene chloride using  a continuous  liquid-
     liquid extractor for aqueous samples for 24 hours.  A separatory funnel
     and 3 x 60 mL methylene chloride may also be used  for aqueous  samples.
     After appropriate cleanup, 10 uL of a tridecane solution of the recovery
     standard (  C12~l>2,3,4-TCDD) are added to the extract  which is then
     concentrated to a final volume of 10 uL.  One to three  uL of the concen-
     trated extract is injected into a gas chromatograph  with a  capillary
     column interfaced to a high-resolution mass spectrometer capable of  rapid
     multiple ion monitoring at resolutions  of at least 10,000 (10  percent
     valley).

     Identification of 2,3,7,8-TCDD is based on the detection of the ions n/z
     319.897 and 321.894 at the same GC retention time  and within -1 to +3
     seconds GC retention time of the internal standard masses of m/z 331.937
     and 333.934.  Confirmation of 2,3,7,8-TCDD (and of other TCDD  isomers) is


                                      D-l

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     based on the ion m/z 258.930 which results from loss  of  COCL by the parent
     molecular ion.

3.   DEFINITIONS
     3.1  Concentration calibration solutions  — solutions  containing known
                                 (unlabeled 2,3,7,8-TCDD),  the  internal  standard
                                 the recovery  standard    C,2-l,2,3,4-TCDD;
          they are used to determine instrument  response  of the analyte
          relative to the internal standard and  of  the  internal standard
          relative to the recovery standard.
amounts of the analyte
13C12-2,3,7,8-TCDD and *v~ -	— '-----•-—' 1J'
     3.2  Field blank — a portion of soil/sediment  or  water  uncontaminated  with
          2,3,7,8-TCDD and/or other TCDDs.

     3.3  Rinsate — a portion of solvent used  to rinse sampling  equipment;  the
          rinsate is analyzed to demonstrate that samples  have  not  been  contami-
          nated during sampling.

                               11
     3.4  Internal standard —   C12-2,3,7,8-TCDD, which is added to every
          sample (except the blank described in Sections 4.2.1  of Exhibit E)
          and is present at the same concentration in every method  blank and
          quality control sample.  It is added  to the soil/sediment  or aqueous
          sample before extraction and is used  to measure  the concentration  of
          each analyte.  Its concentration  is measured  in  every sample,  and
          percent recovery is determined using  an internal standard  method.

     3.5  Recovery standard —   C12~l,2,3,4-TCDD which is added  to every sample
          extract (except for the blank discussed in Sections 4.2.1,  Exhibit  E)
          just before the final concentration step and  HRGC-HRMS  analysis.

     3.6  Laboratory method blank — this blank is prepared in  the  laboratory
          through performing all analytical procedures  except addition of a
          sample aliquot to the extraction  vessel.

     3.7  GC column performance check mixture — a mixture containing known
          amounts of selected standards; it is  used  to  demonstrate  continued
          acceptable performance of the capillary column,  i.e., separation
          (£ 25% valley) of 2,3,7,8-TCDD isomer from all other  21 TCDD isomers,
          and to define the TCDD retention  time window.

     3.8  Performance evaluation sample —  a soil, sediment or  aqueous sample
          containing a known amount of unlabeled 2,3,7,8-TCDD and/or other
          TCDDs.  It is distributed by the  EMSL-LV to potential contractor lab-
          oratories 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 ("blind")
          QC sample in any sample batch submitted to a  laboratory for analysis.

     3.9  Relative response factor — response  of the mass spectrometer  to a
          known amount of an analyte relative to a known amount of  an internal
          standard.
                                      D-2

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     3.10 Mass resolution check — standard method used to demonstrate  static
          resolution of 10,000 minimum (10% valley definition).

     3.11 Positive response for a blank — defined as a signal in the TCDD
          retention time window, at any of the masses monitored,  which  is
          equivalent to or above the method quantitation limit (2 ppt for  soil
          and sediment, and 20 ppq for aqueous samples).

     3.12 Sample rerun — extraction of another 10-g soil or sediment sample
          portion or 1-L aqueous sample, followed by extract cleanup and
          extract analysis.

     3.13 Extract reanalysis — analysis of another aliquot of th final extract.

4.   INTERFERENCES

     Chemicals which elute from the GC column within ^10 scans of the internal
     and/or recovery standard (m/z 331.937 and 333.934) and which produce  within
     the TCDD retention time window ions at any of the masses used to detect  or
     quantify TCDD are potential interferences.  Most frequently  encountered
     potential interferences are other sample components that are extracted
     along with TCDD, e.g. PCBs, chlorinated methoxybiphenyls, chlorinated
     hydroxydiphenylethers, chlorinated benzylphenylethers, chlorinated naphtha-
     lenes, DDE, DDT, etc.  The actual incidence of interference  by these
     chemicals depends also upon relative concentrations, mass spectrometric
     resolution, and chromatographic conditions.  Because very low levels  of
     TCDDs must be measured, the elimination of interferences is  essential.
     High-purity reagents and solvents must be used and all equipment must be
     scrupulously cleaned.  Blanks (Exhibit E, Quality Control,  Section 4) must
     be analyzed to demonstrate absence of contamination that would interfere
     with TCDD measurement.  Column chromatographic procedures are used to
     remove some coextracted sample components; these procedures  must be
     performed carefully to minimize loss of TCDDs during attempts to increase
     their concentration relative to other sample components.

5.   SAFETY

     The toxicity or carcinogen!city of each reagent used in this method has
     not been precisely defined; however, 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 for 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 identi-
     fied O~3) (page D-21).  2,3,7,8-TCDD has been identified as a suspected
     hunan or mammalian carcinogen.  The laboratory is responsible for  ensuring
     that safe handling procedures are followed.
                                      D-3

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6.   APPARATUS AND EQUIPMENT

     6.1  High-Resolution Gas Chromatograph/High-Resolution Mass
          Spectrometer/Data System (HRGC/HRMS/DS)

          6.1.1   The GC must be equipped  for  temperature  programming,  and  all
                 required accessories must be available,  such as  syringes,  gases,
                 and a capillary column.   The GC injection  port must be  designed
                 for capillary columns.   The  use of splitless injection  tech-
                 niques is recommended.   On-column  injection techiques can  be
                 used but this may severely reduce  column lifetime  for
                 noncheraically bonded columns.   When using  the method  in this
                 protocol, a 2-uL injection volume  is used  consistently.  With
                 some GC injection ports,  however,  1-uL injections  may produce
                 improved precision and  chromatographic separation.  A 1-  to 3-uL
                 injection volume may be  used if adequate sensitivity  and
                 precision can be achieved.

          NOTE:   If 1 uL or 3 uL is used  at all as  injection volume, the injec-
                 tion volumes for all extracts, blanks, calibration solutions
                 and the performance check sample must be 1  uL or 3 uL.

          6.1.2   Gas Chromatograph-Mass  Spectrometer Interface

                 The GC-MS interface may  include enrichment  devices, such  as a
                 glass jet separator or a silicone  membrane  separator, or  the
                 gas chromatograph can be directly  coupled  to the mass spectrome-
                 ter ion source.  The interface may include  a diverter valve
                 for shunting the column  effluent and isolating the mass spec-
                 trometer ion source. All components of  the interface should
                 be glass or glass-lined  stainless  steel.  The interface com-
                 ponents should be compatible with  300°C  temperatures.   The
                 GC/MS interface must be  appropriately designed so  that  the
                 separation of 2,3,7,8-TCDD from the other  TCDD isoraers  which
                 is achieved in the gas  chromatographic column is not  appreci-
                 ably degraded.  Cold spots and/or  active surfaces  (adsorption
                 sites) in the GC/MS interface can  cause  peak tailing  and  peak
                 broadening.  It is recommended that the  GC  column  be  fitted
                 directly into the MS ion source.   Graphite  ferrules should be
                 avoided in the GC injection  port since they may  adsorb  TCDD.
                 Vespel" or equivalent ferrules are recommended.

          6.1.3   Mass Spectrometer

                 The static resolution of the instrument  must be  maintained at
                 a minimum 10,000 (10 percent valley). The  mass  spectrometer
                 must be operated in a selected ion monitoring (SIM) mode  with
                 total cycle time (including  voltage reset  time)  of one  second
                 or less (Section 8.3.4.1).  At a minimum,  the following ions
                 which occur at these masses  must be monitored:   m/z 258.930,
                 319.897, 321.894, 331.937 and 333.934.


                                      D-4

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          6.1.4  Data System

                 A dedicated hardware or data system is employed to control the
                 rapid multiple ion monitoring process and to acquire the data.
                 Quantification data (peak areas or peak heights) and SIM traces
                 (displays of intensities of each m/z being monitored as a
                 function of time) must be acquired during the analyses.
                 Quantifications may be reported based upon computer-generated
                 peak areas or upon measured peak heights (chart recording).

NOTE:  Detector zero setting must allow peak-to-peak measurement of the noise
       on the base line.

     6.2  GC Columns

          For isomer-specific determinations of 2,3,7,8-TCDD, the following
          fused silica capillary columns are recommended:  a 60-m SP-2330 (SP-
          2331) column and a 50-m CP-Sil 88 column.  However, any capillary
          column which separates 2,3,7,8-TCDD from all other TCDDs may be used
          for such analyses, but this separation must be demonstrated and
          documented.  Minimum acceptance criteria must be determined per
          Section 8.1.  At the beginning of each 12-hour period (after mass
          resolution has been demonstrated) during which sample extracts or
          concentration calibration solutions will be analyzed, column operating
          conditions must be attained for the required separation on the column
          to be used for samples.  Operating conditions known to produce accept-
          able results with the recommended columns are shown in Table 2 at the
          end of this Exhibit.

     6.3  Miscellaneous Equipment

          6.3.1  Nitrogen evaporation apparatus with variable flow rate.

          6.3.2  Balance capable of accurately weighing to +0.01 g.

          6.3.3  Centrifuge capable of operating at 2,000 rpm.

          6.3.4  Water bath — equipped with concentric ring cover and capable
                 of being temperature-controlled within +2°C.

          6.3.5  Stainless steel spatulas or spoons.

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

          6.3.7  Glove box.

          6.3.8  Drying oven.

     6.4  Glassware

          6.4.1  Soxhlet apparatus — all-glass, Kontes 6730-02 or equivalent;

                                      D-5

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                 90 mm x 35 mm glass thimble;  500-mL flask;  condenser  of  appro-
                 priate size.

          6.4.2  Kuderna-Danish apparatus — 500-mL evaporating flask,  10-mL
                 graduated concentrator tubes  with  ground-glass stoppers,  and
                 3-ball macro  Snyder column (Kontes K-570001-0500,  K-503000-
                 0121  and K-569001-0219 or equivalent).

          6.4.3  Mini-vials — 1-mL borosilicate glass  with  conical-shaped
                 reservoir and screw caps lined  with Teflon-faced silicone disks.

          6.4.4  Funnels — glass;  appropriate size to  accommodate  filter
                 paper used to filter jar extract (volume  of approximately 170 mL)

          6.4.5  Separatory funnel  — 2000 mL  with  Teflon  stopcock.

          6.4.6  Continuous liquid-liquid extractors equipped with  Teflon or
                 glass connecting joints and stopcocks  requiring no lubrication
                 (Hershberg-Wolf Extractor - Ace Glass  Company, Vineland,  NJ;
                 P/N 6841-10 or equivalent).

          6.4.7  Chromatographic columns for the silica  and  alumina chroma-
                 tography — 1 cm ID x 10 cm long and 1  cm ID x 30  cm  long.

          6.4.8  Chromatographic column for the  Carbopak cleanup — disposable
                 5-mL  graduated glass pipets,  6  to  7 mm ID.

          6.4.9  Desiccator.

          6.4.10 Glass rods.

          NOTE:  Reuse of glassware should be  minimized  to avoid the risk of
                 cross contamination.  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 then with
                 high-purity acetone and hexane. 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°C for 15 to 30 minutes.  Volumetric glassware
                 must  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  two cases, rinsing  with high-
                 purity acetone and hexane may be substituted for muffle-furnace
                 heating.  After the glassware is dry and  cool, rinse  with hexane,
                 and store inverted or capped  with  solvent-rinsed aluminum foil
                 in a  clean environment.

7.   REAGENTS AND STANDARD SOLUTIONS

     7.1  Column Chromatography Reagents

                                      D-6

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     7.1.1  Alumina,  acidic — extract the alumina  in a  Soxhlet with
            methylene chloride for 6 hours (minimum of 3  cycles per hour)
            and activate it by heating in a foil-covered glass container
            for 24 hours at 190°C.

     7.1.2  Silica gel — high-purity grade,  type 60,  70-230 mesh; extract
            the silica gel in a Soxhlet with  methylene chloride for 6 hours
            (minimum of 3 cycles per hour) and activate  it  by heating in  a
            foil-covered glass container for  24 hours  at 130°C.

     7.1.3  Silica gel impregnated with 40 percent  (by weight) sulfuric
            acid — add two parts (by weight) concentrated  sulfuric 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.

     7.1.4  Sulfuric acid, concentrated —ACS grade,  specific gravity 1.84.

     7.1.5  Graphitized carbon black (Carbopack C or equivalent), surface
            of approximately 12 m^/g, 80/100  mesh — mix thoroughly 3.6
            grams Carbopak C and 16.4 grams Celite  545®  in  a 40-mL vial.
            Activate at 130°C for six hours.   Store in a desiccator.

     7.1.6  Celite 545®, reagent grade, or equivalent.

7.2  Membrane filters or filter paper with pore size of  <25 urn; rinse with
     hexane before use.

7.3  Glass wool, silanized — extract with methylene chloride and hexane
     and air-dry before use.

7.4  Desiccating Agents

     7.4.1  Sodium sulfate — granular, anhydrous;  before use, extract it
            with methylene chloride for 6 hours (minimum of 3 cycles per
            hour) and dry it for >4 hours in  a shallow tray placed  in an
            oven at 120°C.  Let it cool in a  desiccator.

     7.4.2  Potassium carbonate—anhydrous, granular;  use as such.

7.5  Solvents — high purity, distilled in glass:  methylene chloride,
     toluene, benzene, cyclohexane, methanol, acetone, hexane; reagent
     grade:  tridecane.

7.6  Concentration calibration solutions (Table 1)  — four  tridecane
     solutions containing   C^"! f2,3,4-TCDD  (recovery standard)  and
     unlabeled 2,3,7,8-TCDD at varying concentrations, and  ^c   _2,3,7,8-
     TCDD (internal standard, CAS RN 80494-19-5) at a constant concentration
     must be used to calibrate the instrument.  These concentration  calibra-
     tion solutions must be obtained from the Quality Assurance Division,
     US EPA, Environmental Monitoring Systems Laboratory (EMSL-LV),  Las
     Vegas, Nevada.  However, additional secondary  standards may  be  obtained

                                 D-7

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     from commercial sources, and solutions may be prepared in the con-
     tractor laboratory.  Traceability of standards must  be verified
     against EPAsupplied standard solutions.  Such procedures will be
     documented by laboratory SOPs as required in IFB  Pre-award Bid Con-
     firmations, part 2.f.(4).  It is the responsibility  of the laboratory
     to ascertain that the calibration solutions received are indeed  at the
     appropriate concentrations before they are injected  into the instrument!

NOTE:  Serious overloading of the instrument may occur if the concentration
       calibration solutions intended for a low-resolution MS are injected
       into the high-resolution MS.

     7.6.1  The four concentration calibration solutions  contain unlabeled
            2,3,7,8-TCDD and labeled  13C12~1,2,3,4-TCDD at  nominal concen-
            trations of 2.0, 10.0, 50.0,  and 100 pg/uL, respectively,  and
            labeled   C^2~2,3,7,8-TCDD at a constant nominal concentration
7.7
     7.6.2
       of 10.0 pg/uL.

       Store the concentration calibration solutions in 1-raL mini-
       vials at 4°C.
Column performance check mixture — this solventless mixture  must  be
obtained from the Quality Assurance Division,  Environmental Monitoring
Systems Laboratory, Las Vegas, Nevada,  and dissolved by the Contractor
in 1 mL tridecane.  This solution will  then contain the following
components [including TCDDs (A) eluting closely to 2,3,7,8-TCDD, and
the first- (F) and last-eluting (L) TCDDs when using the columns
recommended in Section 6.2] at a concentration of 10 pg/uL of each of
these isomers:
          Analyte
     Unlabeled 2,3,7,8-TCDD

     13C12-2,3,7,8-TCDD

     1,2,3,4-TCDD (A)

     1,4,7,8-TCDD (A)

     1,2,3,7-TCDD (A)

     1,2,3,8-TCDD (A)

     1,3,6,8-TCDD (F)

     1,2,8,9-TCDD (L)
                                      Approximate Amount  Per Ampule

                                                10'ng

                                                10 ng

                                                10 ng

                                                10 ng

                                                10 ng

                                                10 ng

                                                10 ng

                                                10 ng
7.8  Sample fortification solution — an isooctane solution containing
     the internal standard at a nominal concentration of 10 pg/uL.
                                 D-8

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     7.9  Recovery standard spiking solution — a tridecane solution contain-
          ing the recovery standard at a nominal concentration of 10 pg/uL.
          Ten uL of this solution will be spiked into each sample extract
          (except for the fortified field blank A) before the final concentration
          step and HRGC/HRMS analysis.  It is also used for the dilution of  the
          extracts from samples with high TCDD levels (Section 13.3, Exhibit D).


     7.10 Internal standard spiking solution — a tridecane solution containing
          the internal standard (  C12~2,3,7,8-TCDD) at a nominal concentration  of
          10 pg/uL.  Ten uL of this solution will be added to a fortified field
          blank extract (Section 4.2.1.1, Exhibit E).  This is the only case
          where   C,22,3,7,8-TCDD is used for recovery purposes.

     7.11 Field blank fortification solutions — isooctane solutions containing
          the following TCDD isomers:

          Solution A:  10.0 pg/uL of unlabeled 2,3,7,8-TCDD
          Solution B:  10.0 pg/uL of unlabeled 1,2,3,4-TCDD.

8.   SYSTEM PERFORMANCE CRITERIA

     System performance criteria are presented below.  The laboratory may use
     any of the recommended columns described in Section 6.2.  It must be
     documented that all applicable system performance criteria specified in
     Sections 8.1, 8.2, 8.3 and 8.5 have been met before analysis of any sample
     is performed.  Table 2 provides recommended conditions that can be used to
     satisfy the required criteria.  Table 3 provides a typical 12-hour analysis
     sequence.  The GC column performance and mass resolution checks must be
     performed at the beginning and end of each 12-hour period of operation.

     8.1  GC Column Performance

          8.1.1  Inject 2 uL (Section 6.1.1) of the column performance check
                 solution (Section 7.7) and acquire selected ion monitoring
                 (SIM) data for m/z 258.930, 319.897, 321.894, 331.937 and
                 333.934 within a total cycle time of <1 second (Section
                 8.3.4.1).

          8.1.2  The chromatographic peak separation between 2,3,7,8-TCDD and
                 the peaks representing any other TCDD isomers must be resolved
                 with a valley of <25 percent, where

                                Valley Percent  =  (x/y)(100)


                 x  =  measured as in Figure 1

                 y  =  the peak height of 2,3,7,8-TCDD.

                 It is the responsibility of the laboratory to verify the con-
                 ditions suitable for the appropriate resolution of 2,3,7,8-TCDD

                                      D-9

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            from all other TCDD isomers.  The column performance check
            solution also contains the TCDD isomers eluting first and last
            under the analytical conditions specified in this protocol
            thus defining the retention time window for total TCDD determi-
            nation.  The peaks representing 2,3,7,8-TCDD and the first and
            the last eluting TCDD isomer must be labeled and identified as
            such on the chromatograms (F and L, resp.).  Any individual
            selected ion current profile or the reconstructed total ion
            current (m/z 259 + m/z 320 + m/z 322) constitutes an acceptable
            form of data presentation.

8.2  Mass Spectrometer Performance

     8.2.1  The mass spectrometer must be operated in the electron (impact)
            ionization mode.  Static resolving power of at least 10,000
            (10 percent valley) must be demonstrated before any analysis
            of a set of samples is performed (Section 8.2.2).  Static
            resolution checks must be performed at the beginning and at
            the end of each 12-hour period of operation.  However, it is
            recommended that a visual check (i.e., not documented) of the
            static resolution be made using the peak matching unit before
            and after each analysis.

     8.2.2  Chromatography time for TCDD may exceed the long-term mass
            stability of the mass spectrometer and thus mass drift correc-
            tion is mandatory.  A reference compound [high-boiling
            perfluorokerosene (PFK) is recommended] is introduced into the
            mass spectrometer.  An acceptable lock mass ion at any mass
            between m/z 250 and m/z 334 (m/z 318.979 from PFK is recommended)
            must be used to monitor and correct mass drifts.

     NOTE:  Excessive PFK may cause background noise problems and contami-
            nation of the source resulting in an increase in "downtime"
            for source cleaning.

            Using a PFK molecular leak, tune the instrument to meet the
            minimum required resolving power of 10,000 (10% valley) at
            m/z 254.986 (or any other mass reasonably close to m/z 259).
            Calibrate the voltage sweep at least across the mass range o/z
            259 to m/z 334 and verify that m/z 330.979 from PFK (or any
            other mass close to m/z 334) is measured within +5 ppn (i.e.,
            1.7 mmu, if m/z 331 is chosen) using m/z 254.986 as a reference.
            Documentation of the mass resolution must then be accomplished
            by recording the peak profile of the PFK reference peak m/z
            318.979 (or any other reference peak at a mass close to m/z
            320/322).  The format of the peak profile representation must
            allow manual determination of the resolution, i.e., the hori-
            zontal axis must be a calibrated mass scale (amu or ppn per
            division).  The result of the peak width measurement (performed
            at 5 percent of the maximum which corresponds to the 10%
            valley definition) must appear on the hard copy and cannot
            exceed 100 ppn (or 31.9 nmu if m/z 319 is the chosen reference
            ion).
                                 D-10

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8.3  Initial Calibration

     Initial calibration is required  before  any  samples  are  analyzed  for
     2,3,7,8-TCDD.   Initial calibration  is also  required  if  any routine
     calibration does not meet the required  criteria  listed  in Section 8.6.

     8.3.1  All concentration calibration  solutions listed in Table 1 must
            be utilized for the initial  calibration.

     8.3.2  Tune the instrument with  PFK as  described  in Section  8.2.2.

     8.3.3  Inject  2 uL of the column performance  check  solution  (Section
            7.7) and acquire SIM mass spectral data for  m/z  258.930,
            319.897, 321.894, 331.937 and  333.934  using  a total cycle time
            of ^ 1  second (Section 8.3.4.1). The  laboratory must not
            perform any further analysis until it  has  been demonstrated
            and documented that the criterion listed  in  Section 8.1.2 has
            been met.

     8.3.4  Using the same GC (Section 8.1)  and  MS (Section  8.2)  conditions
            that produced acceptable  results with  the  column performance
            check solution, analyze a 2-uL aliquot of  each of the 4 concen-
            tration calibration solutions  in triplicate  with the  following
            MS operating parameters.

            8.3.4.1  Total cycle time for  data acquisition must be ^  1
                     second.  Total cycle  time includes  the  sum of all the
                     dwell times and  voltage reset times.

            8.3.4.2  Acquire SIM data for  the following  selected
                     characteristic ions:

                       m/z          Compound

                     258.930        TCDD - COC1

                     319.897        Unlabeled TCDD

                     321.894        Unlabeled TCDD

                     331.937        13C12-2,3,7,8-TCDD,  13C12-1,2,3,4-TCDD

                     333.934        13C12-2,3,7,8-TCDD,  13C12~1,2,3,4-TCDD


            8.3.4.3  The ratio of integrated ion current for m/z  319.897 to
                     m/z 321.894 for 2,3,7,8-TCDD  must be between 0.67 and
                     0.90.
            1.3.4.4  The ratio of integrated ion current for m/z  331.937  to
                     m/z 333.934 for 13C12-2,3,7,8-TCDD and 13C12~1,2,3,4-
8,

         TCDD must be between 0.67 and 0.90.

                     D-ll

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                 8.3.4.5  Calculate the relative response factors for unlabeled
                          2,3,7,8-TCDD [RRF(D] relative to * C,2-2,3,7,8-TCDD
                          and for labeled 13C12-2,3,7,8-TCDD [RRF(II)] relative
                          to   C12-1,2,3,4-TCDD as follows:
'12

                  A.
                                       RRF(I)  =  —
                                      RRF(II)  =
                                                  QX ' AIS


                                                  Ais * QRS
                                                  QIS * ARS

where

     Ax  =  sun of the Integrated ion abundances of m/z 319.897 and m/z 321.894
            for unlabeled 2,3,7,8-TCDD.

     AJS  =  sum of the integrated ion abundances of m/z 331.937 and m/z 333.934
            for 13C12-2,3,7,8-TCDD.

    A^g  =  sum of the integrated ion abundances for m/z 331.937 and n/z
            333.934 for 13C12-1,2,3,4-TCDD.

    QIS  =  quantity of 13C12~2,3,7,8-TCDD injected (pg).

    QRS  =  quantity of   C}2~1,2,3,4-TCDD injected (pg).

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

    RRF is a dimensionless quantity;  the units used to express QJS» QRS an(* QX
must be the same.

                 8.3.4.6  Calculate the four means (RRFs) and their respective
                          relative standard deviations (%RSD) for the response
                          factors from each of the triplicate analyses for both
                          unlabeled and 13C12-2,3,7,8-TCDD (Form H-2).

                 8.3.4.7  Calculate the grand means RRF(I) and RRF(II) and their
                          respective relative standard deviations (%RSD) using
                          the four mean RRFs (Section 8.3.4.6) (Form H-2).

                 8.3.4.8  Calculate the routine calibration permissible range
                          for RRF(I) and RRF(II) using a ^20% window from the
                          grand means RRF(I) and RRF(II) (Section 8.3.4.7)
                          (Form H-2).

     8.4  Criteria for Acceptable Calibration

          The criteria listed below for acceptable calibration must be met
          before analysis of any sample is performed.

                                     • D-12

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     8.A.I  The percent relative standard deviation (RSD)  for  the  response
            factors from each of the  triplicate  analyses  for both  unlabeled
            and   Cj2~2,3,7,8-TCDD must be less  than 20 percent.
                                                               1 «1
     8.4.2  The variation of the 4 mean RRFs  for unlabeled and  C^-
            2,3,7,8-TCDD obtained from the triplicate analyses must be
            less than 20 percent RSD.

     8.4.3  SIM traces for 2,3,7,8-TCDD must  present a signal-to-noise
            ratio of ^2.5 for m/z 258.930, m/z 319.897 and, m/z 321.894.

     8.4.4  SIM traces for   Cj2~2»3,7,8-TCDD must  present a signal-to-
            noise ratio >2.5 for m/z  331.937  and m/z 333.934.

     8.4.5  Isotopic ratios (Sections 8.3.4.3 and 8.3.4.4) must be within
            the allowed range.

     NOTE:  If the criteria for acceptable calibration listed  in Sections
            8.4.1 and 8.4.2 have been met, the RRF  can be  considered inde-
            pendent of the analyte quantity for  the calibration concentra-
            tion range.  The mean RRF from 4  triplicate determinations  for
            unlabeled 2,3,7,8-TCDD and for 13C12-2,3,7,8-TCDD  will be  used
            for all calculations until routine calibration criteria (Section
            8.6) are no longer met.  At such  time,  new mean RRFs will  be
            calculated from a new set of four triplicate  determinations.

8.5  Routine Calibrations

     Routine calibrations must be performed at  the  beginning of a  12-hour
     period after successful mass resolution  and GC column performance
     check runs.

     8.5.1  Inject 2 uL of the concentration  calibration  solution  which
            contains 10 pg/uL of unlabeled 2,3,7.8-TCDD,  10.0  pg/uL
            of 13C12-2,3,7,8-TCDD and 10 pg/uL    C12~l,2,3,4-TCDD.
            Using the same GC/MS/DS conditions  as used in Sections 8.1,
            8.2 and 8.3, determine and document  acceptable calibration  as
            provided in Section 8.6.

8.6  Criteria for Acceptable Routine  Calibration

     The following criteria must be met before  further analysis is per-
     formed.  If these criteria are not met,  corrective action must be
     taken and the instrument must be recalibrated.

     8.6.1  The measured RRF for unlabeled 2,3,7,8-TCDD must be within 20
            percent of the mean values established  (Section 8.3.4.8) by
            triplicate analyses of concentration calibration solutions.

     8.6.2  The measured RRF for 13C12-2,3,7,8-TCDD must  be within 20  per-
            cent of the mean value established  by triplicate analysis
            of the concentration calibration  solutions (Section 8.3.4.8).

                                 D-13

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          8.6.3  Isotopic ratios (Sections 8.3.4.3  and 8.3.4.4)  must be within
                 the allowed range.

          8.6.4  If one of the above criteria is not  satisfied,  a second attempt
                 can be made before  repeating the entire  initialization process
                 (Section 8.3).

          NOTE:  An initial calibration must be carried out whenever the HRCC  2
                 solution is replaced by a new one  from a different  lot.

9.   QUALITY CONTROL PROCEDURES

     See Exhibit E for QA/QC requirements.

10.  SAMPLE PRESERVATION AND HANDLING

     10.1 Chain-of-custody procedures — see Exhibit  G.

     10.2 Sample Preservation

          10.2.1 When received,  each soil or sediment 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 the temperature is maintained  between 25° and 35°C.
                 After a portion of  a sample has been removed for analysis,
                 return the remainder of the sample to its original  container
                 and store as stated above.

          10.2.2 Each aqueous sample will be contained in a 1-liter  glass
                 bottle.  The bottles with the samples are stored at 4°C in a
                 refrigerator located in a locked limited-access area.

          10.2.3 To avoid photodecomposition, protect samples from light.

     10.3 Sample Handling

     CAUTION:  Finely divided soils  and sediments contaminated with  2,3,7,8-TCDD
               are hazardous because of the potential for inhalation 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).

          10.3.1 Pre-extraction  sample treatment

                 10.3.1.1 Horaogenization — Although  sampling personnel will
                          attempt to collect homogeneous  samples, the contrac-
                          tor shall  examine each sample and judge if it needs
                          further mixing.

                 NOTE:  Contractor personnel have the responsibility to take  a
                        representative sample portion; this responsibility

                                      D-14

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                        entails efforts to make the sample  as  homogeneous  as
                        possible.  Stirring is recommended  when  possible.

                 10.3.1.2 Centrifugation — When a soil  or  sedimerit  sample
                          contains an obvious liquid phase,  it must  be
                          centrifuged to separate the liquid from the solid
                          phase.  Place the entire sample in a suitable  centri-
                          fuge bottle and centrifuge for 10 minutes  at 2000 rpn.
                          Remove the bottle from the centrifuge.   With a dis-
                          posable pipet, remove the liquid  phase and discard
                          it.  Mix the solid phase with  a stainless  steel
                          spatula and remove a portion to be weighed and analyzed.
                          Return the remaining solid portion to  the  original
                          sample bottle (which must be empty)  or to  a clean,
                          empty sample bottle which is properly  labeled, and
                          store it as described in 10.2.1.

                 CAUTION:  The removed liquid may contain TCDD and should  be
                           disposed as a liquid waste.

                 10.3.1.3 Weigh between 9.5 and 10.5 g of the  soil or sediment
                          sample ^+0.5 g) to 3 significant  figures.  Dry it to
                          constant weight at 100eC.  Allow  the sample to cool
                          in a desiccator.  Weigh the dried soil to  3 signifi-
                          cant figures.  Calculate and report  percent moisture
                          on Form H-9.

11.  SAMPLE EXTRACTION

     11.1 Soil/Sediment Extraction

          11.1.1 Immediately before use, the Soxhlet apparatus is charged
                 with 200 to 250 mL benzene which is then refluxed for 2 hours.
                 The apparatus is allowed to cool, disassembled  and  the  benzene
                 removed and retained as a blank for later  analysis  if required.

          11.1.2 Accurately weigh to 3 significant figures  a 10-g (9.50  g  to
                 10.50 g) portion of the wet soil or sediment  sample.  Mix 100
                 uL of the sample fortification solution (Section 7.8) with
                 1.5 mL acetone (1000 pg of   Cj2~2>3,7,8-TCDD)  and  deposit the
                 entire mixture in small portions on several sites on the
                 surface of the soil or sediment.

          11.1.3 Add 10 g anhydrous sodium sulfate and mix  thoroughly using a
                 stainless steel spoon spatula.

          11.1.4 After breaking up any lumps, place the  soil-sodium  sulfate
                 mixture in the Soxhlet apparatus using  a glass  wool plug  (the
                 use of an extraction thimble is optional). Add 200 to  250 mL
                 benzene to the Soxhlet apparatus and reflux for 24  hours. The
                 solvent must cycle completely through the  system at least 3
                 times per hour.

                                      D-15

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     11.1.5 Transfer the extract to a Kuderna-Danish apparatus and
            concentrate to 2 to 3 mL.  Rinse the column and flask with 5  mL
            benzene and collect the rinsate in the concentrator tube.
            Reduce the volume in the concentrator tube to 2 to 3 mL.
            Repeat this rinsing and concentrating operation twice more.
            Remove the concentrator tube from the K-D apparatus and  care-
            fully reduce the extract volume to approximately 1 mL with a
            stream of nitrogen using a flow rate and distance such that
            gentle solution surface rippling is observed.

      NOTE:  Glassware used for more than one sample must be carefully
             cleaned between uses to prevent cross-contamination (Note on
             page D-6).

11.2  Extraction of Aqueous Samples

      11.2.1  Mark the water meniscus on the side of the 1-L sample  bottle
              for later determination of the exact sample volume.   Pour
              the entire sample (approximately 1 L)  into a 2-L separatory
              funnel.

      11.2.2  Mix 100 uL of the sample fortification solution with 1.5 mL
              acetone (1000 pg of ^€^"2,3,7,8-TCDD)  and add the mixture
              to the sample in the separatory funnel.

      NOTE:  A continuous liquid-liquid extractor may be used in place of
             a separatory funnel.

      11.2.3  Add 60 raL methylene 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.
              Allow the organic layer to separate from the water phase for
              a minimum of 10 minutes.  If an emulsion interface between
              layers exists, the analyst must employ mechanical techniques
              (to be described in the final report)  to complete the  phase
              separation.  Collect the methylene chloride (3 x 60 mL)
              directly into a 500-mL Kuderna-Danish  concentrator (mounted
              with a 10-mL concentrator tube) by passing the sample  extracts
              through a filter funnel packed with a  glass wool plug  and 5
              g of anhydrous sodium sulfate.  After the third extraction,
              rinse the sodium sulfate with an additional 30 mL of methylene
              chloride to ensure quantitative transfer.

     11.2.4   Attach a Snyder column and concentrate the extract until
              the apparent volume of the liquid reaches 1 mL.  Remove  the
              K-D apparatus and allow it to drain and cool for at least
              10 minutes.  Remove the Snyder column, add 50 mL benzene,
              reattach the Snyder column and concentrate to approximately
              1 mL.  Rinse the flask and the lower joint with 1 to 2 mL
              benzene.  Concentrate the extract to 1.0 mL under a gentle
              stream of nitrogen.
                                 D-16

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     11.2.5   Determine the original  sample  volume by  refilling  the sample
              bottle to the mark  and  transferring the  liquid to  a 1000-mL
              graduated cylinder.   Record  the  sample volume to the nearest
              5 raL.

11.3 Cleanup Procedures

     11.3.1 Prepare  an acidic  silica  column  as follows:  Pack a  1 cm x 10
            cm chromatographic  column with a glass wool plug, a  layer (1
            cm) of Na2S04/K2C(>}(l :1) , 1.0  g  silica gel (Section  7.1.2) and
            4.0 g of 40-percent w/w sulfuric acid-impregnated silica gel
            (Section 7.1.3).  Pack  a  second  chromatographic column (1 cm x
            30 cm) with a glass wool  plug, 6.0 g  acidic alumina  (Section
            7.1.1) and top with a 1-cm  layer of sodium sulfate (Section
            7.4.1).   Add hexane to  the  columns until they  are free of
            channels and air bubbles.

     11.3.2 Quantitatively transfer the benzene extract (1 tnL) from the
            concentrator tube  to  the  top of  the silica gel column.  Rinse
            the concentrator tube with  two 0.5-mL portions of hexane.
            Transfer the rinses to  the  top of  the silica gel column.

     11.3.3 Elute the extract  from  the  silica  gel column with 90 mL hexane
            directly into a Kuderna-Danish concentrator.   Concentrate the
            eluate to 0.5 mL,  using nitrogen blow-down as  necessary.

     11.3.4 Transfer the concentrate  (0.5 mL)  to  the top of the  alumina
            column.   Rinse the K-D  assembly  with  two 0.5-mL portions of
            hexane and transfer the rinses to  the top  of the alumina
            column.   Elute the alumina  column  with 18  mL hexane  until the
            hexane level is just  below  the top of the  sodium sulfate.
            Discard  the eluate.  Columns must  not be allowed to  reach
            dryness  (i.e., a solvent  "head"  must  be maintained.)

     11.3.5 Place 30 mL of 20-percent (v/v)  methylene  chloride in hexane
            on top of the alumina and elute  the TCDDs  from the column.
            Collect  this fraction in a  50-mL Erlenmeyer  flask.

     11.3.6 Prepare  an 18-percent Carbopak C/Celite 545® mixture by thoroughly
            mixing 3.6 grams Carbopak C (80/100 mesh)  and  16.4 grams Celite
            545® in  a 40-mL vial.   Activate  at 130°C for 6 hours.  Store
            in a desiccator.  Cut off a clean  5-mL disposable glass pipet
            (6 to 7mm ID) at the  4-mL mark.  Insert a  plug of glass wool
            (Section 7.3) and  push  to the  2-mL mark.   Add  340 to 600 mg of
            the activated Carbopak/Celite  mixture (see NOTE) followed by
            another  glass wool plug.  Using  two glass  rods, push both
            glass wool plugs simultaneously  towards the Carbopak/Celite
            mixture  and gently compress the  Carbopak/Celite plug to a
            length of 2 to 2.5 cm.   Preelute the  column with 2 mL toluene
            followed by 1 mL of 75:20:5 methylene chloride/methanol/benzene,
            1 mL of  1:1 cyclohexane in  methylene  chloride, and 2 mL hexane.
            The flow rate should  be less than  0.5 mL/min.  While the

                                 D-17

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                 column is still wet  with hexane,  add  the  entire  eluate  (30 mL)
                 from the alumina column (Section  11.3.5)  to  the  top  of  the
                 column.  Rinse the Erlenmeyer flask which contained  the extract
                 twice with 1  mL hexane and add the rinsates  to the top  of the
                 column.  Elute the column sequentially with  two  1-mL aliquots
                 hexane, 1 mL  of 1:1  cyclohexane in methylene chloride,  and 1
                 mL of 75:20:5 methylene chloride/ methanol/benzene.   Turn the
                 column upside down and elute  the  TCDD fraction with  6 mL tolu-
                 ene into a concentrator tube.   Warm the tube to  approximately
                 60°C and reduce the  toluene volume to approximately  1 mL using
                 a stream of nitrogen.   Carefully  transfer the concentrate into
                 a 1-mL mini-vial and,  again at elevated temperature,  reduce the
                 volume to about 100  uL using  a stream of  nitrogen.   Rinse the
                 concentrator  tube with 3 washings using 200  uL of 1% toluene
                 in CH2C12. Add 10 uL  of the  tridecane solution  containing the
                 recovery standard and  store the sample in a  refrigerator until
                 HRGC/HRMS analysis is  performed.

                 NOTE:  The amount of activate Carbopak/Celite mixture required
                        to form a 2-to  2.5-cm  plug in  the  column  depends  on the
                        density of the  Celite  being used.

12.  ANALYTICAL PROCEDURES

     12.1  Remove the sample extract or  blank from  storage  and allow it to warm
          to ambient laboratory temperature.  With a stream of dry, purified
          nitrogen, reduce the extract/blank volume to 10  uL.

     12.2  Inject a 2-uL aliquot of the  extract  into the GC, operated  under the
          conditions previously used  (Section  8.1) to  produce acceptable  results
          with the performance check  solution.

     12,3  Acquire SIM data according  to 12.3.1. Use the same acquisition and
          MS operating conditions previously used  (Section 8.3.4) to  determine
          the relative response factors.

          12.3.1 Acquire SIM data for the following selected  characteristic ions:

                   m/z                        Compound

                 258.930                      TCDD - COC1

                 319.897                      Unlabeled TCDD
                                                   *
                 321.894                      Unlabeled TCDD

                 331.937                      13C12-2,3,7,8-TCDD, 13C,2-1,2,3,4-
                                              TCDD

                 333.934                      13C12-2,3,7,8-TCDD, 13C12-1,2,3,4-
                                              TCDD
                                      D-18

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          NOTE:  The acquisition period must at least encompass  the  TCDD  reten-
                 tion time window previously determined  (Section 8.1.2, Exhibit
                 D).

     12.4 Identification Criteria

          12. A.I The retention time (RT) (at maximum peak height)  of the  sample
                 component m/z 319.897 must be within -1 to +3  seconds  of the
                 retention time of the peak for the isotopically labeled  internal
                 standard at m/z 331.937 to attain a positive identification of
                 2,3,7,8-TCDD.  Retention times of other tentatively identified
                 TCDDs must fall within the RT window established by analyzing
                 the column performance check solution (Section  8.1).   Retention
                 times are required for all chroma tograms .

          12.4.2 The ion current responses for m/z 258.930,  319.897  and 321.894
                 must reach maximum simultaneously (_+ 1  sec), and  all ion
                 current intensities must.be > 2.5 times noise  level for
                 positive identification of a TCDD or group of coeluting  TCDD
                 isomers.

          12.4.3 The integrated ion current at m/z 319.897 must  be between 67
                 and 90 percent of the ion current response at m/z 321.894.

          12.4.4 The integrated ion current at m/z 331.937 must  be between 67
                 and 90 percent of the ion current response at m/z 333.934.

          12.4.5 The integrated ion currents for m/z 331.937 and 333.934  must
                 reach their maxima within +_ 1 sec.

          12.4.6 The recovery of the internal standard *^Cj2~2»3, 7,8-TCDD must
                 be between 40 and 120 percent.

13.   CALCULATIONS

     13.1 Calculate the concentration of 2,3,7,8-TCDD (or any other  TCDD  isomer
          or group of coeluting TCDD isomers) using the  formula:
                                  cx
                                           AX
                                       AIS *  W *  RRF(I)
where:
     Cx  =  unlabeled 2,3,7,8-TCDD (or any other unlabeled TCDD isomer or group of
            coeluting TCDD isomers) concentration in pg/g.

     Ax  =  sum of the integrated ion abundances determined for m/z 319.897
            and 321.894.

         =  sum of the integrated ion abundances determined for m/z 331.937
            and 333.934 of r3C12-2,3, 7,8-TCDD (IS = internal standard).

                                      D-19

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     QIS =  quantity (in picograms) of   C12~2,3,7,8-TCDD added to the
            sample before extraction (Qjg = 1000 pg).

      W  =  weight (in grams) of dry soil or sediment sample or volume of
            aqueous sample converted to grams.

 RRF(I)  =  calculated m<
            ean relative response factor for unlabeled 2,3,7,8-TCDD
              C12-2,3,7,8-TCDD.  This represents the grand mean of
the RRF(I)'s obtained in Section 8.3.4.5.
            relative to    12
     13.2 Calculate the recovery of the internal standard   C12~2,3,7,8-TCDD
          measured in the sample extract, using the formula:
          Internal standard             Ajg
          percent recovery    =  Y	  • 100
                                   ARg * RRF(II)
Where:
             sun of the integrated ion abundances determined for m/z 331.937
             and 333.934 of 13C12-2,3,7,8-TCDD (IS  =  internal standard).
     ARS  "  sum °f c^e integrated ion abundances determined for m/z 331.937
             and 333.934 of   C12-l ,2,3,4-TCDD (RS  =  recovery standard).

       Y  =  0.1 for the "10-yL extract" injection (to be reported on Forms H-l,
             H-5 and H-9).

   and y  =  1.2 for the "24-yL extract" injection (Section 13.3) (to be reported
             on Form H-9 used for reporting the diluted extract analysis).

— —                                                              1 3
RRF(II)  =  calculated mean relative response factor for labeled   C|2-2, 3,7,8-
            TCDD relative to   C12-l ,2 ,3,4-TCDD.  This represents the grand
            mean of the RRF(II)'s calculated in Section 8.3.4.5.

     13.3  If the concentration of the most abundant TCDD isomer (or group of
           coeluting TCDD isomers) exceeds 100 pg/uL in the 10 uL final extract,
           the linear range of response vs. concentration may have been exceeded,
           and a diluted aliquot of the original sample extract must be analyzed.
           Accurately dilute 2 uL of the remaining original extract with 22 uL
           of the tridecane solution containing 10 pg/uL of the recovery standard
           (Section 7.9, Exhibit D).

     13.4  Total TCDD concentration — all positively identified isomers of TCDD
           must be within the RT window and meet all identification criteria
           listed in Sections 12.4.2 and 12.4.3.  Use the expression in Section
           13.1 to calculate the concentrations of the other TCDD isomers, with
           Cx becoming the concentration of any unlabeled TCDD isomer or group
           of coeluting TCDD isomers.

C Total TCDD  =  Sum of the concentrations of the individual TCDDs including
                 2,3,7,8-TCDD.
                                      D-20

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     13.5  Estimated Detection Limit — For samples in which no unlabeled
           2,3,7,8-TCDD was detected, calculate the estimated minimum detectable
           concentration.  The background area is determined by integrating the
           ion abundances for n/z 319.897 and 321.894 in the appropriate region
           of the selected ion current profiles, multiplying that  area by 2.5,
           and relating the product area to an estimated concentration that
           would produce that product area.

           Use the formula:

                                          (2.5) ' (Ax) ' (QIS)
                                         (AIS) •  (RRF(I)) •  (W)

where

     CE  =  estimated concentration of unlabeled 2,3,7,8-TCDD required to
            produce Ax.

     Ax  =  sum of integrated ion abundances for m/z 319.897 and 321.894 in the
            same group of >5 scans used to measure
            sum °^ integrated ion abundances for the appropriate ion character-
            istic of the internal standard, m/z 331.937 and m/z 333.934.

   , RRF(I), and W retain the definitions previously stated in Section 13.1.
Alternatively, if peak height measurements are used for quantification,  measure
the estimated detection limit by the peak height of the noise in the 2,3,7,8-
TCDD RT window.

     13.6  The relative percent difference (RPD) is calculated as follows:

                        I  Si - S2 |           I  Si - S2 |
             RPD  -  	  «=  	  x  100
                     Mean Concentration      (S^ + S2>/2

          Sj and 82 represent sample and duplicate sample results.

References

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

2.   "OSHA Safety and Health Standards, General Industry" (29 CFR1910),
     Occupational 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.


                                      D-21

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TABLE 1.  COMPOSITION OF CONCENTRATION CALIBRATION  SOLUTIONS


HRCC1
HRCC2
HRCC3
HRCC4
Recovery Standard
13C12-1,2,3,4-TCDD
2.0 pg/uL
10.0 pg/uL
50.0 pg/uL
100.0 pg/uL
Analyte
2,3,7,8-TCDD
2.0 pg/uL
10.0 pg/uL
50.0 pg/uL
100.0 pg/uL
Internal Standard
13C12-2,3,7,8-TCDD
10.0 pg/uL
10.0 pg/uL
10.0 pg/uL
10.0 pg/uL
               Sample Fortification  Solution

             10.0 pg/uL of 13C12-2,3,7,8-TCDD
             Recovery Standard  Spiking  Solution

               10.0  pg/uL 13C12-1,2,3,4-TCDD



            Field Blank Fortification Solutions

          A)  10.0 pg/uL of unlabeled 2,3,7,8-TCDD

          B)  10.0 pg/uL of unlabeled 1,2,3,4-TCDD
             Internal Standard  Spiking  Solution

              10 pg/uL of 13C12-2,3,7,8-TCDD
             (Used only in Section 4.2.1.1, Exhibit E)
                             D-22

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                 TABLE 2.  RECOMMENDED GC OPERATING  CONDITIONS
Column coating

Film thickness

Column dimensions

Helium linear velocity


Initial temperature

Initial time

Temperature program
Approximate 2,3,7,8-TCDD
retention time
SP-2330 (SP-2331)

0.2 urn

60 m x 0.24 mm

28-29 cm/sec
at 240°C

150°C

4 min

Rapid increase to 200°C
(15°C/min)
200°C to 250°C
at 4°C/min

27 min
CP-SIL 88

0.22 urn

50 m x 0.22 mm

28-29 cm/sec
at 240°C

200°C

1 min

Program from 200°C
to 240°C
at 4°C/min


22 min
         TABLE 3.  TYPICAL 12-HOUR SEQUENCE  FOR 2,3,7,8-TCDD ANALYSIS
1. Static mass resolution check and mass
measurement error determination
2. Column performance check
3 . HRCC2
4. Sample 1 through Sample "N"
5. Column performance check
6. Static mass resolution check
10/20/84
10/20/84
10/20/84
10/20/84
10/20/84
10/20/84
0700h
0730h
0800h
0830h
1800h
1830h
                                      D-23

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     100-1
       80-
CO
g  60H
o
    d>
    OC
       20-
                00
                (O
                CO
                   (F)
                                                                                    (U
                       24:00
                                  26:00
28:00
30:00
32:00
                                               Time
Figure 1.
              Selected ion current profile for m/z 322 produced by MS analysis of performance check

              solution using a 60-m SP-2331  fused silica capillary column and  conditions listed in
              Table 2.

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    EXHIBIT E
QA/QC Requirements

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SUMMARY OF QC ANALYSES

o  Initial and periodic calibration and instrument performance checks.

o  Field blank analyses (Section A.I); a minimum of one fortified field blank
   pair shall be analyzed with each sample batch;  an additional fortified field
   blank pair must be analyzed when a new lot of absorbent and/or solvent is used.

o  Analysis of a batch of samples with accompanying QC analyses:

        Sample Batch —<2b samples, including field blank and rinsate  sample(s).

        Additional QC analyses per batch:


        Fortified field blanks              2

        Method blank                       (1*)

        Duplicate sample                    1

                              TOTAL         3(A)


        * A method blank is required whenever a fortified field blank shows a
          positive response as defined in Section 3.11, Exhibit D.

o  "Blind" OC samples may be submitted to the contractor as ordinary soil,
   sediment or water samples included among the batch of samples.  Blind samples
   include:

        Uncontaminated soil, sediment and water,

        Split samples,

        Unidentified duplicates, and

        Performance evaluation samples.

QUALITY CONTROL

1.   Performance Evaluation Samples — Included among the samples in all batches
     will be samples containing known amounts of unlabeled 2,3,7,8-TCDD and/or
     other TCDDs that may or may not be marked as other-than-ordinary samples.

2.   Performance Check Solutions

     2.1  At the beginning of each 12-hour period during which samples  are to
          be analyzed, an aliquot each of the 1) GC column performance  check
          solution and 2) high-resolution concentration calibration solution
                                      E-l

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     No. 2 (HRCC2) shall be analyzed to demonstrate adequate GC resolution
     and sensitivity, response factor reproducibility,  and mass range
     calibration.  A mass resolution check shall also be performed to
     demonstrate adequate mass resolution using an appropriate
     reference compound (PFK is recommended).

     These procedures are described in Section 8 of Exhibit D.  If the
     required criteria are not met, remedial action oust be taken before
     any samples are analyzed.

2.2  To validate positive sample data, the GC column performance check
     and the mass resolution check must be performed also at the end of
     each 12-hour period during which samples are analyzed.

     2.2.1  If the contractor laboratory operates only  during one period
            (shift) each day of 12 hours or less, the GC performance check
            solution must be analyzed twice (at the beginning and end of
            the period) to validate data acquired during the interim
            period.  This applies also to the mass resolution check.

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

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

2.4  Deviations from criteria specified for the GC performance check or
     for the mass resolution check (Section 8, Exhibit  D) invalidate all
     positive sample data collected between analyses of the performance
     check solution, and the extract from those positive samples shall be
     reanalyzed Exhibit C).

The GC column performance check mixture, concentration  calibration solu-
tions, and the sample fortification solutions are to be obtained from the
EMSL-LV.  However, if not available from the EMSL-LV, standards can be
obtained from other sources, and solutions can be prepared in the contractor
laboratory.  Concentrations of all solutions containing unlabeled 2,3,7,8-
TCDD which are not obtained from the EMSL-LV must be verified by comparison
with the unlabeled 2,3,7,8-TCDD standard solution (concentration of 7.87
ug/raL) that is available from the EMSL-LV.  When a lower-concentration
standard solution becomes available from the EMSL-LV, it will be substituted
for the 7.87 ug/mL standard.
                                 E-2

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4.   Blanks
     4.1  A method blank Is required whenever a positive  response (Section  3.11,
          Exhibit D) is obtained for a fortified field blank.   To that  effect,
          perform all steps detailed 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,  but
          omit addition of the soil, sediment or aqueous  sample portion.

                                                                  13
          4.1.1  The method blank must contain the same amount  of   C]2~2,3,7,8-
                 TCDD that is added to samples before extraction.

          4.1.2  An acceptable method blank exhibits no positive response (Section
                 3.11, Exhibit D) for any of the characteristic ions monitored.
                 If the method blank which was extracted  along  with a batch of
                 samples is contaminated, all positive samples  must be  rerun
                 (Exhibit C).

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

                 4.1.2.2  If new batches  of reagents or solvents contain
                          interfering contaminants,  purify or discard them.

     4.2  Field blanks — Each batch of samples contains  a field blank  sample
          of uncontaminated soil/sediment or water that is to be fortified
          before analysis according to Section 4.2.1, Exhibit E.  In addition
          to this field blank, a batch of samples may include a rinsate,  that
          is a portion of solvent (usually trichloroethylene) that was  used to
          rinse sampling equipment.  The  rinsate is  analyzed to assure  that the
          samples have not been contaminated by the  sampling equipment.

          4.2.1  Fortified field blank pair

                 4.2.1.1  Fortified field blank A:  2,3,7,8-TCDD

                 4.2.1.1.1  Weigh a 10-g  portion or  use 1 liter (for aqueous
                            samples) of the specified field blank sample  and
                            add 100 uL of the solution containing 10.0  pg/uL of
                            2,3,7,8-TCDD  (Table 1, Exhibit D) diluted in 1.5 mL
                            of acetone (Section 11.1.2, Exhibit D).

                 4.2.1.1.2  Extract using the procedures  beginning in Sections
                            11.1 or 11.2  of Exhibit  D, as applicable, add 10 uL
                            of the Internal standard solution (Section  7.10,
                            Exhibit D) and analyze a 2-uL aliquot of the con-
                            centrated extract.
                                      E-3

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       NOTE:  This is the only case where the recovery standard is
              used for other than recovery purposes.

       4.2.1.1.3  Calculate the concentration (Section 13.1, Exhibit
                  D) of 2,3,7,8-TCDD and the percent recovery of
                  unlabeled 2,3,7,8-TCDD.  If the percent recovery at
                  the measured concentration of 2,3,7,8-TCDD is <40
                  percent or >120 percent, report the results and
                  repeat the fortified field blank extraction and
                  analysis with a second aliquot of the specified
                  field blank sample (Exhibit C).

       4.2.1.1.4  Extract and analyze a new fortified simulated field
                  blank whenever new lots of solvents or reagents are
                  used for sample extraction or for column chromato-
                  graphic procedures.  When a fortified simulated
                  field blank produces a positive response (Section
                  3.11, Exhibit D) for any m/z being monitored at the
                  retention time of 1,2,3,4-TCDD, a method blank
                  (Section 4.1, Exhibit E) is required.

       NOTE:  For this purpose only, the Contractor will simulate
              field blanks by using clean sand or distilled water.

       4.2.1.2  Fortified field blank B:  1,2,3,4-TCDD

       4.2.1.2.1  Repeat steps 4.2.1.1.1 to 4.2.1.1.3 using unlabeled
                  1,2,3,4TCDD (instead of 2,3,7.8-TCDD) and 13C
                  12-1,2,3,4-TCDD (instead of 13C12-2,3,7,8-TCDD) as
                  recovery standard.

       4.2.1.2.2  Extract and analyze a new fortified simulated field
                  blank whenever new lots of solvents or reagents are
                  used for sample extraction or for column chromato-
                  graphic procedures.  When a fortified simulated
                  field blank produces a positive response (Section
                  3.11, Exhibit D) for any m/z being monitored at the
                  retention time of 2,3,7,8-TCDD, a method blank
                  (Section 4.1, Exhibit E) is required.

4.2.2  Rinsate sample

       4.2.2.1  The rinsate sample must be fortified as a regular
                sample.

       4.2.2.2  Take a 100-mL aliquot of sampling equipment rinse
                solvent (rinsate sample), filter, if necessary, and
                 Id 100 uL of the solution containing 10.0 pg/uL of
                 >C12-2,3,7,8-TCDD (Table 1, Exhibit D).
                            E-4

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                 4.2.2.3  Using a Kuderna-Danish apparatus,  concentrate to
                          approximately 5 mL.

                 A.2.2.4  Transfer the 5-mL concentrate in 1-mL portions to a  1-
                          mL mini-vial, reducing the volume  as  necessary with  a
                          gentle stream of dry nitrogen;  see Exhibit D,
                          Section 11.1.5 for volume reduction procedures.

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

                 4.2.2.6  Just before analysis, add 10 uL tridecane recovery
                          standard spiking solution (Table 1, Exhibit D), and
                          reduce the volume to a final volume of 10 uL (no
                          column chromatography is required).

                 4.2.2.7  Analyze an aliquot following the same procedures used
                          to analyze samples (Section 12, Exhibit D).

                 4.2.2.8  Report percent recovery of the internal standard and
                          the level of contamination by any  TCDD isomer (or
                          group of coeluting TCDD Isomers) on Form H-5 in pg/mL
                          of rinsate solvent.
5.   Duplicate Analyses

     5.1  Laboratory duplicates — in each batch of samples, locate the sample
          specified for duplicate analysis and analyze a second 10-g soil or
          sediment sample portion or 1-L water sample.

          5.1.1  The results of laboratory duplicates (percent  recovery and
                 concentrations of 2,3,7,8-TCDD and total TCDD) must agree
                 within 50 percent relative difference (difference expressed as
                 percentage of the mean).  If the relative difference is >50
                 percent, the Contractor shall immediately contact the Sample
                 Management Office for resolution of the problem.  Report all
                 results.

          5.1.2  Recommended actions to help locate problems:

                 5.1.2.1  Verify satisfactory instrument performance
                          (Section 8, Exhibit D).

                 5.1.2.2  If possible, verify that no error  was made while
                          weighing sample portions.

                 5.1.2.3  Review the analytical procedures with the performing
                          laboratory personnel.

6.   Percent Recovery of the Internal Standard 13C,2~2,3,7,8-TCDD — For each
     sample, method blank and rinsate, calculate the
     13.2, Exhibit D) of the measured concentration of
                                      E-5
percent recovery (Section
lf   Cl2-2,3,7,8-TCDD.  If

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     the percent recovery is <40 percent or >120 percent for a sample, analyze
     a second portion of that sample and report both results (Exhibit C).

NOTE:  A low or high percent recovery for a blank does not require discarding
       analytical data but it may indicate a potential problem with future
       analytical data.

7.   Identification Criteria

     7.1  If either of the two identification criteria (Sections 12.4.1 and
          12.4.2, Exhibit D) is not met, it is reported that the sample does
          not contain unlabeled 2,3,7,8-TCDD at the calculated detection limit
          (Section 13.5, Exhibit D).

     7.2  If the first two initial identification criteria are met, but the
          third, fourth, fifth or sixth criterion (Sections 12.4.3 through
          12.4.6, Exhibit D) is not met, that sample is presumed to contain
          interfering contaminants.  This must be noted on the analytical
          report form and the sample must be rerun or the extract reanalyzed.
          Detailed sample rerun and extract reanalysis requirements are
          presented in Exhibit C.

8.   Blind QC Samples — Included among soil, sediment and aqueous samples may
     be QC samples that are not specified as such to the performing laboratory.
     Types that may be included are:

          8.1    Uncontaminated soil, sediment or water.

                 8.1.1  If a false positive is reported for such a sample,
                        the Contractor shall be required to rerun the entire
                        associated batch of samples (Section 2.3.3, Exhibit C).

          8.2    Split samples — composited sample portions sent to more  than
                 one laboratory.

          8.3    Unlabeled field duplicates — two portions of a composited
                 sample.

          8.4    Performance evaluation sample — soil/sediment or water sample
                 containing a known amount of unlabeled 2,3,7,8-TCDD and/or
                 other TCDDs.

                 8.4.1  If the performance evaluation sample result falls
                        outside the acceptance windows established by EPA, the
                        Contractor shall be required to rerun the entire associ-
                        ated batch of samples (Exhibit C).

                 NOTE: EPA acceptance windows are based on previously generated
                       data.

9.   Records - At each contractor laboratory, records must be maintained on


                                      E-6

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     site for six months after contract completion to document the quality of
     all data generated during the contract performance.   Before any records are
     disposed, written concurrence from the Contracting Officer must be obtained.

10.  Unused portions of samples and sample extracts must  be preserved for
     six months after sample receipt; appropriate samples may be selected
     by EPA personnel for further analyses.

11.  Reuse of glassware is to be minimized to avoid the risk of contamination.

LABORATORY EVALUATION PROCEDURES

1.   On a quarterly basis, the EPA Project Officer and/or designated
     representatives 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 dioxin 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

     2.1  The PE sample set will be sent to a participating laboratory to
          verify the laboratory's continuing ability to produce acceptable
          analytical results.  The PE sample will be representative of the
          types of samples that will be subject to analysis under this contract.

     2.2  When the PE sample results are received, they are scored using the
          PE Sample Score Sheet shown in Figure 1.  If a false positive
          (e.g., a PE sample not containing 2,3,7,8-TCDD and/or other TCDDs
          but reported by the laboratory to contain it and/or them) is reported,
          the laboratory has failed the PE analysis requirement.  The Project
          Officer will notify the laboratory immediately if such an event
          occurs.

     2.3  As a general rule, a laboratory should achieve 75 percent or more of
          the total possible points for all three categories, and 75 percent  or
          more of the maximum possible points in each category to be considered
          acceptable for this program.  However, the Government reserves the
          right to accept scores of less than 75 percent.

     2.4  If unanticipated difficulties with the PE samples are encountered,
          the total points may be adjusted by the Government evaluator in an
          impartial and equitable manner for all participating laboratories.
                                      E-7

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     Number of          Maximum Possible        Recommended Passing
     PE Samples             Score                  Score (75%)
        1                    290                       218

        2                    475                       356

        3                    660                       495

        4                    845                       634

        5                   1030                       773


On-Site Laboratory Evaluation

3.1  An on-site laboratory evaluation is performed to verify that (1)  the
     laboratory is maintaining the necessary minimum level in instrumen-
     tation and levels of experience in personnel committed to the con-
     tract and (2) that the necessary quality control/quality 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.
     Photographs may be taken during the on-site laboratory evaluation
     tour.

3.2  The sequence of events for the on-site evaluations is shown in
     Figure 2.  The Site Evaluation Sheet (SES) (Figure 3) is used to
     document the results of the evaluation.
                                    E-8

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Laboratory
                     PERFORMANCE EVALUATION SAMPLE SCORE SHEET

                                                         Date
                                                               False Positive
     False Positive - If a laboratory reports a false
     positive on any PE sample, the laboratory may be
     disqualified, i.e., rendered ineligible for
     contract award based on the failure to pass the
     PE sample analysis requirement.

     2,3,7,8-TCDD

     Other TCDD(s)
                                                               (  ) Yes  (  )  No

                                                               (  ) Yes  (  )  No
                                                            Possible
                                                              Score
II.  Calibration Data

     1.  Method Blank:

         a.  Results properly recorded on Forms H-l,  H-5 and
             H-9.

         b.  No native TCDD isomers at /or above method
             quantitative limit.

         c.  Results documented by selected ion
             monitoring (SIM) traces for m/z being
             monitored to detect TCDDs.

         d.  Percent recovery of 13C,2-2,3,7 ,8-TCDD
                 and  120%.
                                                                5


                                                                5
     2.  Initial Concentration Calibration:

         a.  Results properly recorded on Forms H-2             5
             and H-8.

         b.  The percent relative standard deviation
             (RSD) for the response factors for each
             of the triplicate analyses for both unlabeled
             and   C12-2,3,7,8-TCDD less than 20%.              5

         c.  The variation of the 4 mean RRFs for both
             unlabeled and labeled 2,3,7,8-TCDD obtained
             from the triplicate analyses less than 20% RSD.    5

         d.  For unlabeled 2,3,7,8-TCDD the abundance ratio
             must be >Q.67 and £0.90 for m/z 319.897 to
             321.894.                                           5

             Figure 1.  Performance evaluation sample score sheet,

                                      E-9
                                                                        Score
                                                                       Achieved

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                                                   Possible    Score
                                                     Score    Achieved
e.  The abundance ratios must be XK67 and £0.90
    for 331.937 to 333.934 for 13C12~2,3,7,¥-TCDD
    and 13C12-1,2,3,4-TCDD.                            5

f.  Results must be documented with appropriate
    SIM traces, labeled with the corresponding EPA
    sample numbers, and calculations.                  5

 Performance Checks:

a.  GC resolution and MS resolution checks performed
    at the beginning and end of each 12-hour period.   5

b.  Results of performance checks properly recorded
    on Form H-4.                                       5

c.  MS Resolution:  PFK (or alternate) tune shows
    appropriate mass resolution (Section 8.2,
    Exhibit D) with mass assignment accuracy
    within +5 ppm.                                     5

d.  GC Resolution:  chromatograms meet the criteria
    specified in Section 8.1, Exhibit D.               5

Routine Calibration:

a.  Performed each 12 hours, after MS and  GC
    resolution checks,  using HRCC2.                    5

b.  Results of routine calibrations properly
    reported on Forms H-3 and H-8.                     5

c.  For unlabeled 2,3,7,8-TCDD:  abundance
    ratio must be X).67 and <0.90 for m/z
    319.897 to 321.894.                                5

d.  Abundance ratio correct for isotopically  ,
    labeled standards (e.g., 331.937/333.934
    must be X5.67 and <0.90 for 13C12-2,3,7,8-TCDD
    and 13C12-1,2,3,4-TCDD).                           5

e.  Response factors [RRF(I) and RRF(II)]  are
    within ^20% of the mean of the respective
    initial calibration response factors.               5

f.  Signal-to-Noise (S/N) Ratio:  SIM traces
    for 2,3,7,8-TCDD demonstrate S/N of >2.5.           5

g.  Results documented with appropriate SIM
    traces and calculations.                           5
                                        Subtotal II   105

                   Figure 1.  (Continued).

                             E-10

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                                                            Possible     Score
                                                              Score    Achieved
III. Performance Evaluation (PE)  Sample  Data
     (Scores to be determined for each sample
      in the PE set)

     1.  Forms H-l and H-9 properly filled out  for  sample.       5

     2.  Measured concentration of unlabeled
         2,3,7,8-TCDD within acceptance  window
         established  by EPA.                                  40

     3.  Estimated concentration of total TCDDs
         within acceptance window established by
         EPA.                                                 20

     4.  Identification Criteria  for 2,3,7,8-TCDD:

         a.   Retention time (RT)  (at maximum peak
             height)  of the sample component m/z
             319.897  is within -1 to +3  seconds
             of the m/z 331.937 13C122,3,7,8-TCDD
             internal standard peak.                          10

         b.   The ion  current responses for m/z
             258.930, 319.897 and 321.894 must  reach
             a maximum simultaneously (+1 second)
             and must be >2.5 times noise level.               10

         c.   The m/z  319.897/321.894 ratio is X>.67
             and _<0.90.                                       10

         d.   The m/z  331.937/333.934 ratio is X>.67
             and _<0.90.                                         5

         e.   The S/N  ratio for m/z 331.937 and
             333.934  is >2.5.                                   5

     5.  Identification Criteria  for other TCDDs:

         a.   Retention time must  fall into window
             established by GC performance check.                5

         b.   The ion  current responses for m/z
             258.930, 319.897, and 321.894 reach
             a maximum simultaneously (j^l second)
             and are  2^«5 times noise level.                   10

         c.   The m/z  319.897/321.894 ratio is
             20.67 and ^0.90.                                   5

                            Figure 1.  (Continued).
                                      E-ll

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                                                        Possible     Score
                                                          Score     Achieved
 6.  Concentrations of unlabeled TCDDs
     are calculated according to D-13.1.                    10

 7.  Duplicate analysis values agree  within
     ±50%.                                                 10

 8.  Estimated detection limits calculated
     according to D-13.5.                                  10

 9.  Percent recovery of 13C,2~2,3,7,8-TCDD
     >40 and O20%.                                        10

10.  Results documented with appropriate
     SIM traces and calculations.                          20
                                           Subtotal  III    185

                                                  Total    290

                        Figure 1.   (Continued).
                                  E-12

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                EVENT SEQUENCE FOR ON-SITE LABORATORY EVALUATION
I.    Meeting with Laboratory Manager and Project Manager

      Introduction; discuss purpose of visit; discuss problems with data
      submitted by the laboratory.

II.   Verification of Personnel

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

III.  Verification of Instrumentation

      Review equipment in place and committeed to project (Section II,  SES).
      The Contractor must demonstrate adequate equipment redundancy,  as defined
      in SES, Section II.D.,  to ensure his capability to perform the required
      analyses in the required time.

IV.   Quality Control Procedures

      Walk through the laboratory to review:

       1.  Sample receiving and logging procedures,
       2.  Sample and extract storage area,
       3.  Procedures to prevent sample contamination,
       4.  Security procedures for laboratory and samples,
       5.  Safety procedures,
       6.  Conforraance to written SOPs,
       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.  Technical and managerial review of laboratory operations and
             data package preparations,
      13.  Procedures for data handling, analysis, reporting and case
             file preparation, and
      14.  Chain-of-custody procedures.

V.    Review of Standard Operating Procedures (SOPs)

      Review SOPs with the Project Manager to assure that the laboratory under-
      stands the dimensions and requirements of the program.

VI.   Identification of Needed Corrective Actions

      Discuss with the Project Manager the actions needed to correct weaknesses
      identified during the site inspection, PE sample analysis or production of

          Figure 2.  Event Sequence for On-Site Laboratory Evaluation.

                                      E-13

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       reports (hard copies and, if appropriate,  manual calculations)  and documen-
       tation.  Determine how and when corrective actions  will  be  documented,
       how and when improvements will be demonstrated,  and identify the contractor
       employee responsible for corrective actions.

 VII.  Previously Identified Problems

       Check the most recent SES to verify that all  previously  identified
       problems have been corrected.

VIII.  Identification of New Problems

       a.  Discuss any weaknesses identified in the  performance evaluation
           sample analyses and reports.

       b.  Discuss any weaknessess identified in this site inspection.
                                Figure 2.  (Continued)
                                       E-14

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                             SITE EVALUATION SHEET

Laboratory:  	  Date:


Location:
                                   EVALUATORS
              Name                                        Organization

1.
2.
3.
4.
5.
6.
7.

I.    Laboratory Personnel Committed to Project:

      A.  Project Manager (responsible for overall technical  effort)

          Name:
          Title:
      B.  GC/MS Operator:
          Experience:*
                          (one year minimum)
      C.  GC/MS Data Interpreter:
          Experience:*  	
                          (two year minimum)

      D.  Person responsible for sample exraction, column chromatography
            and extract concentration:  	
          Experience:*  	
                          (one year minumum)

      E.  Person(s) responsible for calculations and report preparation:
          Hardcopy Reports:  	
      F.  Person responsible for handling,  storage and (if appropriate)
            preparation of solutions of standard compounds:
*Experience is deemed to mean "more than 50 percent of the person's productive
 work time."

                       Figure 3.  Site Evaluation Sheet.

                                     E-15

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      G.  Person responsible for standards preparation/storage:


      H.  Person responsible for record keeping:


      I.  Quality Assurance Officer: 	

      J.  Personnel checklist
                                                                (  )  Yes  (  )  No
          1.  Do personnel assigned to this project  have
              the appropriate level and type of experience
              to successfully accomplish the objectives of
              this program?

          2.  Is the organization adequately staffed to          (  )  Yes  (  )  No
              meet project requirements in a tmely
              manner?

          3.  Does the Laboratory Quality Assurance  officer     (  )  Yes  (  )  No
              report to senior management levels?

          4.  Was the Quality Assurance officer available       (  )  Yes  (  )  No
              during the evaluation?

II.   Laboratory Equipment

      A.  Gas chromatograph(s)*

          Manufacturer and Model:  	
          Installation Date:
          Type of Capillary Column Injection System:   	
          Capillary Column to be used (length,  ID,  coating,  etc.):
          Necessary Ancillary Equipment (gases,  syringes,  etc.):
      B.  High Resolution Mass Spectrometer( s)*

          Static Resolution Capability (10,000 min.):
          Peak matching system:  	
          Manufacturer and Model:  	
          Installation Date:
          Pertinent Modifications:  	
          Peak Matching System/Accuracy (Mfg. spec.):

      C.  Data System(s)*
          Manufacturer and Model:
* If more than one GC/MS/DC, indicate system 1,2,3,  etc.,  by numbering
  components with 1,2,3, etc.

                            Figure 3.  (Continued).

                                      E-16

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          Installation Date:
          Software Version Identifier:
          Appropriate selected ion monitoring software/hardware  (  )  Yes   (  ) No
          Capability to produce hard copies  of computer-
            generated information                               (  )  Yes   (  ) No

      D.  Evidence that at least one GC/MS/DS system can  be  reasonably
          expected to be operating acceptably at  any given time:

          (  )  More than one adequate GC/MS/DS system is  available in-house,
               (i.e.,meeting requirements  specified  in SOW Section 6.1,
               Exhibit D).

          (  )  Appropriate in-house replacement parts and trained  service
               personnel are available.

          (  )  A service contract is in place with guaranteed response  time
               (specify type of contract and limitations). 	
          (  )  Voltage control devices are used on major instruments;  isolated
               circuits are used.

          (  )  Other (specify)  	

III.   Facilities Checklist

      A.  Does the laboratory appear to have adequate           (  )  Yes  (  )  No
          workspace (120 sq. feet,  6 linear feet of
          unencumbered bench space  per analyst)?

      B.  Does the laboratory have  a source of distilled/       (  )  Yes  (  )  No
          demineralized water?

      C.  Is the analytical balance located away from           (  )  Yes  (  )  No
          draft and areas subject  to rapid temperature
          changes or vibration?

      D.  Has the balance been calibrated within one year       (  )  Yes  (  )  No
          by a certified technician?

      E.  Is the balance routinely  checked with class S         (  )  Yes  (  )  No
          weights before each use and the results recorded
          in a logbook?

      F.  Is the laboratory maintained in a clean and           (  )  Yes  (  )  No
          organized manner?
                            Figure 3.   (Continued).

                                      E-17

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      G.  Is the facility designed for hazardous organic        ( ) Yes  (  )  No
          chemical analysis?

          1.  Is ventilation provided in the sample             ( ) Yes  (  )  No
              preparation areas?

          2.  Are vented hoods available and adequately         ( ) Yes  (  )  No
              vented in the sample preparation areas?

          3.  Are the hoods equipped with charcoal              ( ) Yes  (  )  No
              and HEPA filters?

          4.  Are instruments, including GC/MS pumps,           ( ) Yes  (  )  No
              vented into hoods or control devices such
              as charcoal traps?

      H.  Are adequate secured facilities provided for          ( ) Yes  (  )  No
          storage of samples,  extracts,  and calibration
          standards, including cold storage?

      I.  Are the temperatures of the cold storage units        ( ) Yes  (  )  No
          recorded daily in logbooks?

      J.  Are chemical waste disposal policies/procedures       ( ) Yes  (  )  No
          in place?

      K.  Is the laboratory secure?                             ( ) Yes  (  )  No

IV.   Analysis Control Checklist

      A.  Do the project personnel have SOPs for the required   ( ) Yes  (  )  No
          activities?

      B.  Is a logbook maintained for each instrument and       ( ) Yes  (  )  No
          is information such  as calibration data and
          instrument maintenance continually recorded?

      C.  Do the analysts record bench data in a neat           ( ) Yes  (  )  No
          and accurate manner?

      D.  Standards

          1.  Are fresh analytical standards prepared           ( ) Yes  (  )  No
              at a frequency consistent with good QC?

          2.  Are reference materials properly labeled with     ( ) Yes  (  )  No
              concentrations,  date of preparation, and the
              identity of the person preparing the sample?

          3.  Is a standards preparation and tracking           ( ) Yes  (  )  No
              logbook maintained?

                             Figure 3. (Continued).

                                      E-18

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          4.  Are working standards traceable to EPA            (  )  Yes   (  )  No
              standards or validated against EPA
              standards?

V.    Documentation/Tracking Checklist

      A.  Is a sample custodian designated?  If yes,            (  )  Yes   (  )  No
          name of sample custodian.
          Name:  	

      B.  Are the sample custodian's procedures and              (  )  Yes   (  )  No
          responsibilities documented?  If yes, where
          are these documented?
          Are the chain-of-custody procedures documented?       (  )  Yes  (  )  No

      C.  Are written Standard Operating Procedures (SOPs)       (  )  Yes  (  )  No
          developed for receipt of samples?  If yes, where
          are the SOPs documented (laboratory manual,
          written instructions, etc.)?

      D.  Are quality assurance procedures documented           (  )  Yes  (  )  No
          and available to the analysts?  If yes,  where
          are these documented?

      E.  Are written Standard Operating Procedures (SOPs)       (  )  Yes  (  )  No
          developed for compiling and maintaining  sample
          document files?  If yes, where are the SOPs
          documented (laboratory manual, written
          instructions, etc.)?

      F.  Are the magnetic tapes stored in a secure area?       (  )  Yes  (  )  No

      G.  Are samples that require preservation stored          (  )  Yes  (  )  No
          in such a way as to maintain their integrity?
          If yes, how are the samples stored?

      Documentation/Notebooks Checklist

      A.  Is a permanently bound notebook with preprinted,       (  )  Yes  (  )  No
          consecutively numbered pages being used?

      B.  Is the type of work clearly displayed on the          (  )  Yes  (  )  No
          notebook?

      C.  Is the notebook maintained in a legible  manner?       (  )  Yes  (  )  No

      D.  Are entries noting anomalies routinely recorded?       (  )  Yes  (  )  No

                            Figure 3.  (Continued).

                                      E-19

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      E.  Has the analyst avoided obliterating entries or the   (  )  Yes  (  )  No
          use of a pencil?

      F.  Are inserts (i.e. chromatograms,  computer print-      (  )  Yes  (  )  No
          outs, etc.) permanently affixed to the notebook
          and signed across insert edge and page?

      G.  Has the supervisor of the individual maintaining the  (  )  Yes  (  )  No
          notebook personally examined and reviewed the notebook
          periodically, and signed his/her name therein,  together
          with the date and appropriate comments as to whether or
          not the notebook is being maintained in an appropriate
          manner?

      H.  Where applicable, is the notebook holder              (  )  Yes  (  )  No
          referencing reports or memoranda  pertinent to
          the contents of an entry?

VI.   Quality Control Manual Checklist

      Does the laboratory maintain a Quality Assurance/         (  )  Yes  (  )  No
      Quality Control (QA/QC) Manual?

      Does the manual address the important elements            (  )  Yes  (  )  No
      of a QA/QC program, including the following:

      A.  Personnel                                             (  )  Yes  (  )  No

      B.  Facilities and equipment                              (  )  Yes  (  )  No

      C.  Operation of instruments                              (  )  Yes  (  )  No

      D.  Documentation of Procedures                           (  )  Yes  (  )  No

      E.  Procurement and inventory practices                   (  )  Yes  (  )  No

      F.  Preventive maintenance                                (  )  Yes  (  )  No

      G.  Reliability of data                                   (  )  Yes  (  )  No

      H.  Data validation                                       (  )  Yes  (  )  No

      I.  Feedback and corrective action                        (  )  Yes  (  )  No

      J.  Instrument calibration                                (  )  Yes  (  )  No

      K.  Recordkeeping                                         (  )  Yes  (  )  No

      L.  Internal audits                                       (  )  Yes  (  )  No

                            Figure 3.  (Continued).


                                      E-20

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       Are QA/QC responsibilities and reporting relationships    (  )  Yes  (  )  No
       clearing defined?

       Have standard curves been adequately documented?          (  )  Yes  (  )  No

       Are laboratory standards traceable?                       (  )  Yes  (  )  No

       Are quality control charts maintained for each            (  )  Yes  (  )  No
       routine analysis?

       Do QC records show corrective action when                 (  )  Yes  (  )  No
       analytical results fail to meet QC criteria?

       Do supervisory personnel review the data and QC results?   (  )  Yes  (  )  No

 VII.  Data Handling Checklist

       Are data calculations checked by a second person?        ( ) Yes  (  ) No

       Are data calculations documented?                        ( ) Yes  (  ) No

       Do records indicate corrective action that has           ( ) Yes  (  ) No
       been taken on projected data?

       Are limits of detection determined and reported          ( ) Yes  (  ) No
       properly?

       Are all data and records retained for the                ( ) Yes  (  ) No
       required amount of time?

       Are quality control data (e.g., standard curve           ( ) Yes  (  ) No
       duplicates) accessible for all analytical
       results?

VIII.  Summary

       Do responses to the evaluation indicate that             ( ) Yes  (  ) No
       project and supervisory personnel are aware
       of QA/QC and its application to the project?

       Do project and supervisory personnel place               ( ) Yes  (  ) No
       positive emphasis on QA/QC?

       Have responses with respect to QA/QC aspects of          ( ) Yes  (  ) No
       the project been open and direct?

       Has a cooperative attitude been displayed by all         ( ) Yes  (  ) No
       project and supervisory personnel?
                             Figure 3.  (Continued).

                                       E-21

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Does the organization place the proper emphasis          ( ) Yes  ( ) No
on quality assurance?

Have any QA/QC deficiencies been discussed before        ( ) Yes  ( ) No
leaving?

Is the overall quality assurance adequate to             ( ) Yes  ( ) No
accomplish the objectives of the project?

Have corrective actions recommended during               ( ) Yes  ( ) No
previous evaluations been implemented?

Are any corrective actions required?  If so,             ( ) Yes  ( ) No
list the necessary actions below.
                                E-22

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                                   TECHNICAL REPORT DATA
                            (fleae read Instructions on the reverse before completing)
1. REPORT NO.
                             2.
                                                           3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE                                           6. REPORT DATE
  PROTOCOL FOR THE ANALYSIS OF 2,3,7,8-TETRACHLORODIBENZOt
  p-DIOXIN BY HIGH-RESOLUTION GAS CHROMATOGRAPHY/HIGH-
  RESOLUTION MASS SPECTROMETRY
                                    6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
 J.  S. Stanley and T.
                                                           B. PERFORMING ORGANIZATION REPORT NO.
M. Sack
>. PERFORMING ORGANIZATION NAME AND ADDRESS
 Midwest Research Institute
 425 Volker Boulevard
 Kansas City, Missouri   64110
                                                           10. PROGRAM ELEMENT NO.
                                    11. CONTRACT/GRANT NO.

                                      Contract  Number SAS 1576X
12. SPONSORING AGENCY NAME AND ADDRESS
  Environmental Monitoring  Systems Laboratory - LV, NV
  Office of Research and  Development
  U.S.  Environmental Protection Agency
  Las Vegas, NV 89114
                                                           13. TYPE OF REPORT AND PERIOD COVERED
                                    14. SPONSORING AGENCY CODE

                                    EPA/600/07
15. SUPPLEMENTARY NOTES
  Project Officer - Werner  F.  Beckert, Environmental Monitoring Systems Laboratory
                                                    Las  Vegas,  NV 89114
16. ABSTRACT
       An analytical protocol for the determination  of  2,3,7,8-tetrachlorodibenzo-p-
  dioxin (TCDD) and total  TCDDs in soil, sediment and aqueous samples using high-
  resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS) was
  developed using the best features of several candidate methods and input from experts
  in  the field.  Preliminary  tests led to refinements of the chromatographic cleanup
  procedures and corresponding changes in the protocol.  A final single-laboratory
  evaluation of the refined protocol, consisting of  triplicate analyses of five solid
  and five aqueous samples showed that the method is useful for the determination of
  2,3,7,8-TCDD and total TCDDs at concentrations from 10 to 200 pg/g (ppt) in soils and
  100 to 2,000 pg/L (ppq)  in  aqueous samples.  Based on  the data generated and on the
  evaluation of several options,  parts of the protocol were modified at the EMSL-LV to
  lower the quantitation limit for TCDD to 2 ppt in  soil/sediments and to 20 ppq in
  aqueous samples.
17.
                                KEY WORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
                       b.lDENTIFlERS/OPEN ENDED TERMS  C. COSATI FieW/GlOUp
16. DISTRIBUTION STATEMENT

 RELEASE TO PUBLIC
                       19. SECURITY CLASS (This Report)
                           UNCLASSIFIED
                                                                         21. NO. OF PAGES
                       20. SECURITY CLASS (This page I
                           UNCLASSIFIED
22. PRICE
EPA Pw« 2220.1 («•». 4.77)   PNKVIOUI EDITION if OMOLCTC

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