gy)
                                                                PB88-199526
  Guidelines  for  the Determination of Halogenated
  Dibenzo-p-Dioxins  and  Dibenzofurans in
  Commercial  Products (RE-ANNOUNCEMENT of
  PB88-101050 - see  notes  field for explanation)
  Midwest Research  Inst. ,  Kansas City,  MO
  Prepared  for

  Environmental Protection Agency,  Washington, DC
  Sep 87
EJBD
ARCHIVE
EPA
560-
5-
87-
007
  U.S. DEPARTMENT OF COMMERCE
National Technical Information Service

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               United States          Office of Pesticides      EPA-560/5-87/007
               Environmental Protection     and Toxics Substances     September 1987
               Agency            Washington DC 20460
6-              __	P3d8-199526

§EPA        Guidelines for the

               Determination  of Halogenated

               Dlbenzo-p-Dioxins

               and Dibenzofurans

               in Commercial Products
3:
CD
                  US EPA
          Headquarters and Chemical Libraries
             EPA West Bldg Room 3340
                Mailcode 3404T
             1301 Constitution Ave NW
              Washington DC 20004
                 202-566^556           Permanent Collection
                  REPRODUCED BY

                  U.S. DEPARTMENT OF COMMERCE

                                SerViCe

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                                   TECHNICAL REPORT DATA
                            (Pleau read laurucnara on :ne mint snare eomntetmfi
I  flS.aa«TNC.
 EPA 560/5-87-007
                                                                      AC=sssi
,. TITUS ANO :
Guidelines for the Determination of  Halogenated
Dibenzo-o-dioxins and Dibenzofurans  in  Commercial
   Products             	.^__
                                                            9.
                                                                   3ATS
                                                            a.
                                                                                   :=cs
7. -I.THOHIS)
 Oavid H. Stsale  and Jonn S.  Stanley
                                                           .3.'
                                                             8833A-01
9. "SPOH.vii.NG ORGANISATION .MA.VI6 ANO AOOHESS
 Midwest  Research  Institute
 *25 Volker Boulevard
 Kansas City, MO   5-1110
                                                                      sLsME.N
                                                             63-02-1252
12. jPCNSOfING AG
                   MAMS AAIO AOOHESS
                                                           .13.
 Field Studies  Sranch,  TS-798
 Office of Toxic  Substances
 U.S. Environmental  Protection Agency
 Washington.  DC  20*60	
                                                              Final
                                                           14.S7CNSOPING AG£NCV ;3 = 6
                                                             Office of Toxic iuostances
                                                             U.S. Environ,  '"-ot,
IS. SUPfM-SMBNTAav .NOTHS                T^sie'C  IV
-------
GUIDELINES FOR THE DETERMINATION OF HALOGENATED OIBENZO-fi-OIOXINS
            AND OIBENZOFURANS IN COMMERCIAL PRODUCTS
                               by

                         David H. Staele
                         John S. Stanley
                          FINAL REPORT

                     Work Assignment  No.  33

                EPA Prime Contract  No.  68-02-4252
                    MRI  Project No. 8833-A01



                          June  19,  1987
                           Prepared  for:

               U.S.  Environmental  Protection Agency
             Office  of Pesticides  and Toxic  Substances
                       Field Studies Branch
                        401 M Street, S.W.
                       Washington, DC  20460

          Attn:   j^ Janet C.  Remmers, Work  Assignment Manager
                 Mr. Tom Murray, Work Assignment Manager
                 Dr. Joseph Breen, Program Manager

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                    DISCLAIMER





This  document  has been revised  «*<«««*'?»?";£"*

-------
                                   PREFACE


          This document  provides  guidelines  for  the  determination of haloge-
nated dibenzo-g-dioxins (HDDs) and dibenzofurans (HDFs) in commercial products
using instrumental methods  of  analysis.   This  report includes  a brief review
of analytical  requirements  and existing  methods  for  the analysis of HDDs and
HDFs, guidelines  for  sampling  and instrumental analysis for  these compounds,
and a quality assurance plan.

          The analytical guidelines  focus on high resolution gas chromatog-
raphy (HRGC) with either mass  spectrometry (MS) or electron capture  detectors
(ECD) for qualitative identification and quantisation.  Bioanalytical tech-
niques may be a more cost-effective alternative,  but these procedures are  not
sufficiently validated at this time  to be useful.  Therefore,  guidelines for
bioanalytical methods have not been included in this document.

          Preparation of the document was initiated under MRI Project No.  8201
and completed under MRI Project No. 8800  for the U.S. Environmental  Protection
Agency's (EPA) Office of Toxic Substances, Field  Studies  Branch,  EPA Prime
Cor)^r|ct. ^gs.rs6jQp2-3938 and 68-02-4252,  Work  Assignments  41 and 33, respec-
tively,^.  Tom Murray, Work Assignment Manager*,  and Or.  Joseph  Breen, Project
Officer.  A  draft version  of  this report was  submitted  for EPA review  and
approval on  June  3,  1985,  and a  revised  draft submitted  on June 27, 1985.

          The main body and Appendix A (analytical guidelines) of this document
were prepared by Mr.  David H.  Steele and  Dr.  John S.  Stanley, Work Assignment
Leader, with  input  from  Dr.  Mitchell D.   Erickson, Mr. Richard D. Brown, and
Ms. LeAnn L. Timmons.   Appendices B, C, and D, containing the quality assurance
project plan  and  approaches  for sampling commercial  products  for  analysis,
were prepared  by  Dr.  John Smith of EPA's  Office  of Toxic Substances  Chemical
Regulations Branch.

                                             MIDWEST RESEARCH INSTITUTE
•-P
                                              aul C. Constant
                                             Program Manager
Approved:



                      \_'>*- Co o>" •>
John E.  Going, Director         /
Chemical Sciences Department
                                     n

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                              TABLE OF CONTENTS

                                                                         Page

I.         Introduction	       1

II.        Scope of Problem	       1

               A.   Diversity of Sample Matrices 	       1
               B.   Potential Number of HDD and HDF Isomers	       1
               C.   Potential Interferences	       2

III.   •    General  Analytical Method Considerations	       3

               A.   Sample Extraction and Cleanup	       3
               B.   Detection Method 	       3
               C.   Method Sensitivity 	       a
               0.   Analytical Standards 	       6
               E.   Quality Assurance/Quality Control (QA/QC)	       6

IV.        Existing Methods	       6

               A.   Instrumental Methods 	     11
               B.   Bioanalytical Methods	     11

V.         Proposed Method	     12

Bibliography	     14

Appendix A - Guidelines for the Analysis of Halogenated Dibenzo-
  g-dioxins and Dibenzofurans in Commercial Products	    A-l

Appendix B - Quality Assurance Project Plan for Measurement of
  Halogenated Dibenzo-g-dioxins and Dibenzofurans 	    8-1

Appendix C - Guidance for Sampling Halogenated Oibenzo-g-dioxins
  and Dibenzofurans	'	    C-l

Appendix D - Guidance for a Sequential Approach to the Sampling of
  Halogenated Dibenzo-g-dioxins and Dibenzofurans 	    Q-l

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I.  INTRODUCTION

          The  Environmental  Protection Agency  was  petitioned (October 22,
1984) by  the  Environmental  Defense Fund (EDF)  and  the National Wildlife
Federation (NWF), pursuant to  Section  21  of  the Toxic  Substances Control  Act
(TSCA),  to issue rules and orders to prevent and reduce environmental contam-
ination by halogenated (specifically brominated and/or chlorinated)  dibenzo-
g-dioxins and  dibenzofurans  (HDDs  and HOFs).   The Agency  has reviewed the
Petition and has granted certain aspects.   In addition, there are a number of
relevant Agency  activities,  planned or currently underway, which address  the
Petitioners'  concerns.  The  Agency  feels  that  in many cases  it  is necessary
to gather preliminary information before the Agency can make  regulatory find-
ings.  As part of the preliminary assessments, appropriate analytical metnoa-
ologies must be identified or developed.

          The objective of this work assignment  is  to  identify and/or develoo
appropriate analytical methodologies to quantitata  HOOs and HOFs -in commercial
products.   This  report  identifies  the scope of  the problem (Section II) and
general  considerations for analytical  methods  (Section III) dealing  with  the
determination  of HOOs  and  HOFs.   Existing analytical methods for the deter-
mination of  chlorinated dioxins  and furans are  discussed  (Section IV), and
general  guidelines  for the sampling and analysis of  HDDs and  HOFs  in commer-
cial products  are provided in  Section  V and Appendix A.  A quality assurance
project plan and guidance for  sampling are contained in Appendices 8, C,  and
0.
II.  SCOPE OF PROBLEM

          The development of  analytical  methodologies for the determination
of HOOs and  HOFs  in commercial products is complicated by the diverse range
of sample matrices,  the potentially large  number of brominated, chlorinated,
and mixed brominated/chlorinated  congeners of the HDDs and HOFs,  and by the
potential  interferences which can be expected to be encountered.   The follow-
ing subsections discuss these considerations individually.

     A.   Diversity of Sample Matrices

          It is anticipated  that  commercial  products which will  be analyzed
under this program will range in volatility from chlorinated solvents to neavy
metal salts  and dyes.  Acids,  bases, and neutral compounds with a wide  range
of molecular geometries can be expected.

     B.   Potential Number of HDD and HDF Isomers

          Both the  dibenzodioxin  and  the dibenzofuran molecules have eight
positions  where  bromine  or chlorine can  substitute.   These positions  are
numbered as  shown for  the dibenzo-£-dioxin and dibenzofuran molecules below.

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          The total number  of  unique positional isotners has been calculated
(MRI Interim Report,  "Analytical  Methodologies for Halogenated  Oioxins  and
Dibenzofurans in Commercial  Products, May 1, 1985) at 1,700 HDD and 3,320 HDF
for bromo, chloro and bromo/chloro-substitution.  Within these compounds there
are 351 HDDs and 666  HDFs with substitution  in the 2,3,7,8-positions.   The
breakdown of the exact number  of  positional isomers with degree  of  halogena-
tion are  provided  in  the  general  method  guidelines (Appendix A,  Tables  1 and
2).  Although the number of theoretical isomers is high, the actual number of
HDDs and  HDFs  in commercial products may be limited to relatively few.  The
analyst must first determine the probability of the presence of bromo-,  chloro-,
or bromo/chloro-HODs  and  HDFs  in  specific  commercial  products. - Clearly,  a
chlorinated commercial product that has had no  contact with bromine needs only
to be analyzed for PCDDs and PCDFs.  Likewise,  a brominated material produced
with no  intermediate  chlorinated  materials, needs only to  be  analyzed  for
brominated HDDs  and HDFs  (PBDDs and  PBOFs).   It is estimated that the  bromo/
chloro-commercial products  requiring  analyses of  all  HDDs and HDFs are  very
few.

     C.   Potential Interferences

          Interferences are  expected  to  arise from contaminants  in  solvents,
reagents, and glassware, as well as contaminants (halogenated diphenyl ethers,
halobenzyl phenyl ethers, bis(chlorophenoxy)methanes, and halogenated phenoxy-
phenols) which are coextracted with the HDDs and HDFs.

          Halogenated diphenyl  ethers  yield mass  spectral  fragment  ions  with
exactly the  same mass and number  of  halogen atoms as  the corresponding HDFs.
Buser  (1975) and Mieure  et al. (1977)  have demonstrated that  basic alumina
column chromatography will  separate  the  chlorodiphenyl  ethers  from  the PCDDs
(polychlorinated dibenzo-g-dioxins) and PCDFs  (polychlorinated dibenzofurans).
Likewise, Huckins et  al.   (1978) have demonstrated that a graphitized charcoal
column will  remove the  chlorodiphenyl ether  interferences while  selectively
isolating and concentrating chlorinated dibenzo-p_-dioxins.'

          The  halobenzyl  phenyl ethers may cause  mass spectral  interferences
to HDDs with mass spectrometry selected  ion monitoring (SIM) techniques.   In-
terferences arise with these compounds when using electron capture detectors;
however,  these compounds  are distinguishable  from PCDDs by mass  spectrometry.
These  compounds  can  also be removed from  sample extracts by further cleanup
on  alumina  or  carbon columns.   The  bis(chlorophenoxy)methanes  have similar
gas chromatographic retention  characteristics  to PCDDs.

          The  halogenated phenoxyphenols  can dehydrohalogenate   in the  hot
inlet  of  the gas chromatograph to form HDDs.   This  has been reported for the
chlorinated phenoxyphenols  (Baker  et al. 1981)  and for 6-hydroxy-2,2',4,4',5,5'-
hexabromobiphenyl  (Gardner  et  al.  1979).   It  is expected that analogous  reac-
tions  will occur for  the  mixed bromo/chloro phenoxyphenols".  In  order  to avoid
false  positives, these  species must be  excluded from the final  extracts,  or
the  extracts  must be methylated  to  prevent ring  closure during  injection.

          In addition to interferences caused  by contaminants in the  sample
extracts,  interferences  may be caused by the  HDDs and HDFs themselves.   This

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will occur due  to  the potentially large  number  of isomers and  to  the  ion
clusters produced by the isotopes of bromine and chlorine.  The potential  for
overlap of the  isotope clusters becomes  greater as  the  potential  for mixed
bromo/chloro-HDDs and HDFs increases.  For example, some interferences might
be  anticipated  for determination of the  tetrabromodibenzo-£-dioxin  (TBDD)
using ions m/z  500 and m/z 502.  The interference  may  result  as  contribution
from  dibromopentachlorodibenzofurans or  bromoheptachlorodibenzo-g-dioxins
which exhibit the  same ions  in  the molecular clusters  (Appendix  A,  Table  5).
Again, it should be noted that the probability of this occurring may  be small
and is undoubtedly a concern only for the mixed bromo/chloro-commercial  prod-
ucts.  Since  little  is known regarding the chemical and physical  nature  of
bromo and bromo/chloro-HDDs and HDFs, it  is not clear whether these interfer-
ences will be resolved by HRGC retention time.   The masses and relative aoun-
dances of the ions in the molecular  ion clusters of the HDDs and HDFs are  pre-
sented as part of the method guidelines (Appendix A).  Possible interferences
from the  molecular  ion clusters of  the HDDs and HDFs  are  also presented  in
the method guidelines.


III.  GENERAL ANALYTICAL METHOD CONSIDERATIONS

          The analytical  procedures  necessary for the quantitative measurement
of HDDs and HDFs in commercial products include (1) the quantitative  extraction
or partitioning of the analytes from the commercial product, (2) separation of
the HDDs and HDFs from interferences present in the extract, and (3)  separation
and quantitation of homologous series or  specific congeners by gas chromatog-
raphy with mass spectral  or electron capture detection.  Some general consid-
erations concerning these procedures are discussed below.

     A.   Sample Extraction and Cleanup

          The most significant difference in the analysis of HDDs and HDFs  in
commercial products  in comparison  with environmental and biological  samples
will be  the  extraction and cleanup  procedures.  The  physical  and chemical
properties of environmental  and biological  matrices are typically different
enough from  the properties of the analytes  to  allow  relative  ease  of  separa-
tion.  In contrast, the commercial  products in most cases may be structurally
similar to the  analytes,  complicating the  separation  and  necessitating the
complete removal of the matrix to avoid interferences in the final determina-
tion.

     B.   Detection Method

          Most  of  the analyses of commercial  products for polychlorinated
dioxins (PCDDs), dibenzofurans (PCDFs), polychlorinated biphenyls (PCBs),  and
polybrominated biphenyls (PBBs) that have been reported in-.the literature  have
employed mass spectrometry  (MS) detection.   Mass  spectrometry will provide
both qualitative and  quantitative  data for HDDs and HOFs.  In addition,  its
primary advantage  over other detectors is  provided  by  confirmation of  the
presence of  a specific analyte.  Mass  spectrometry is  a  proven tool for pro-
viding quantitative  information at  concentration levels ranging from micro-
grams per gram to picograms per gram.

-------
          Several options are available for mass spectrometry analyses includ-
ing (1) full mass scan analysis (FMS), which provides identification based on
complete mass spectra of  individual components;  (2)  limited mass  scan  (LMS),
which allows the analyst to monitor small mass ranges, possibly full molecular
ion clusters; and (3) selected ion monitoring (SIM), which permits the analyst
to select specific ions characteristic of the target compounds.  The mass spec-
trometery method  sensitivity  is  increased by decreasing the range of masses
or number of  ions monitored during an  analysis.  Thus the  SIM  method  is  more
sensitive than  the  LMS  or FMS procedures..   Full scan mass spectrometry is
useful for measurements for the 50 ng/g and higher concentration levels, while
the SIM  technique will  allow  measurements as  low as  1-10 pg/g.  Some  caution
must be exercised in approaching the final analysis since the lower detection
levels  require  that sample  extracts  be  relatively  free  of interference.

          Another option  that is available  is high  resolution  mass  spectrom-
etry  (HRMS).  This  approach can be used  with  any of  the techniques  discussed
above.  The advantage of HRMS is that it allows the analyst to lower the con-
tribution of  signal  due to background and  interferences  by monitoring  the
exact masses  of the analytes.  This usually results in an observed increase
of signal to  noise  and corresponding  lower  limit  of detection (LOD).   The
major drawbacks  of  HRMS involve the availability of  instrumentation,  experi-
enced analysts,  cost, and sample turnaround time.

          Electron  capture  detection  (ECO)  has  proven to  be  effective  for
monitoring  PCDD  and PCDF contaminants at the microgram-per-gram (ppm) level
in some  commercial  products  (pentachlorophenol,  Mieure et al.  1977).   It is
expected that this technique may be suitable for the analysis of HDDs and HDFs
in commercial products.   The  ECD analysis technique  is typically  employed on
samples on  which extensive extraction and cleanup procedures have been  per-
formed, producing extracts of acceptable quality for final analysis.

          The electron capture detector  is highly sensitive to compounds with
halogen substitution and provides an attractive, cost-effective procedure com-
pared to mass spectrometry.   However,  this  detector  provides qualitative in-
formation only,  based  on a response at a certain retention time.   The inci-
dence of false  positives will increase as the analyst presses the detection
limit from  the  micrograms-per-gram level  to  the  low  nanograms-per-gram level
(parts-per-million to parts-per-billion  in commercial product).  The decision
to pursue analysis  based on  ECD will require that the analyst comply with a
well-validated  analytical  procedure  and stringent  QA/QC program to minimize
the number  of false positives during sample analysis.

     C.  Method  Sensitivity

          One of the major concerns in using any of the analytical techniques
is the  ability  to achieve the desired  limits  of  detection. . Table  1 provides
a summary of  the reported detection limits for various PCDDs and PCDFs in phen-
oxyalkanoic acid herbicides (Baker et al. 1981).  These detection limits range
from  5  to  500 ng/g  (ppb).  A detection  limit of 50 ng/g can be achieved for
specific isomers of TCDD and TCDF assuming a conservative instrument  sensi-
tivity  (quadrupole  mass spectrometer)  and a 1-uL injection from a  1-mL final
extract of  a  1-g sample.  This detection  limit can be lowered by (1) increas-
ing the  initial   sample  size (~ 10 g),  (2) decreasing the final extract volume

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fable 1.   A Summary of the Reported Analytical Limits of Detection for Polychlorodibeiwo-g-dioxins  (PCDDs)
                  and Polyclilurodibenzofurans (PCOFs) in Phenoxydlkanojc Acid Herbicides
Reference
Storherr et al.
Brenner et al.
(1972)
Vo.jel (1976)
Rdiiiblad et al.
(1077)
Elvidge (1971)
Wool son et al.
(1972)
Edmunds et al.
(1973)
Crunimelt and
SUIil (1973)
Du&er et al.
(1974)
lluckins et al.
(197ft)
Hdppe et al.
(1978)
Pul/hofer (1979)
Cdi el lo et al
Applicability
2.4.5-T
2.4-D. dichlorprop, mecoprop and
2.4.5-T
2.4.5-T
Butoxypropyl esters of 2.4,5-T
and of fenoprop
2.4,5-T, ethylhexyl ester of 2,4,5-T
and formulations containing the
ethylhexyl ester of 2,4,5-T
2.4-0, 2,4-DB. dichlorprop, 2,4,5-T.
fenoprop and dicamba
2.4.5-T. the butyl and octyl
esters of 2,4,5-T and formulations
of the butyl ester of 2.4.5-T
(50% in mineral oil)
Esters of 2.4.5-T and of fenoprop.
Herbicide Orange
2,4,5-T, formulations of esters of
2,4.5-r and of amine salts of 2.4,5-T
Herbicide Orange
Formulations of esters of 2,4,5-T,
Herbicide Orange
2.4.5-T. alky) esters of. 2.4.5-T
and formulations
Fenoprop
Compounds delected
TCDO
TCDO
fCDD
TCDO
TCDD
Di-, tri-. teti-d-,
penta-, hexa-.
liepta. and octa-CDOs
TCOD
TCDO
TCDO
TCUO and penta-CDD
l)i-, tri-, telrd-,
penld- and hexa-CODb,
di-, tri-. letra-.
pciita- and hexa-COl s
rcoo
ICDI)
Liuiil.of detection
(my ky ' of sample)
0.05
0.01
0.03
0.005
0 05 (on 2,4,5-T
content)
0.5 (for any one
of the compounds
delected)
0 OS
0.05
0 001 (on a stan-
dard solution)
0.02
0.01-0.05
0.005 (on 2.4.5-T
content)
0 01
       PC,  lloodless  Kll,  lyltr JC.   1981.   Pestle Sci 12:297-304.

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(~ 20 uL), OP (3) use of a more sensitive (^ 5 pg/uL) mass spectrometer (mag-
netic sector double  focusing  instrument).   By adjusting  these  options,  the
method limit can be lowered to 0.01 to 1 ng/g.

          It should  be  noted  that these detection  limits are determined  by
the sensitivity of the instrument, and that sample interferences may signifi-
cantly increase  the  detection  limit  for  a given matrix.   The working defini-
tion of  limit  of detection,  as defined in EPA methods for the determination
of 2,3,7,8-TCDD  in soil/sediment  and water, requires  that the signal for  the
compound is at a minimum three times the background (signal to noise).

     0.   Analytical Standards

          Present  commercial  sources of  HDD  and HDF analytical  standards
include  Cambridge  Isotope  Laboratories  (Woburn, Massachusetts),  KOR  Isotopes
(Cambridge,  Massachusetts),  Ultra  Scientific (Hope,  Rhode  Island),  and
Wellington Laboratories  (Guelph,  Ontario,  Canada).   Table 2 provides a sum-
mary of  the  current  availability  of  specific  substituted  unlabeled and  mass-
labeled  HDDs and HDFs.   Table 2  is intended  as  a guide  only, as suppliers
may  add  or  remove reference  materials from  their  offerings  at any time.
Availability should be confirmed with the suppliers.

          A limited number of the mixed bromo/chloro analogs of these compounds
are currently available (see Table 2).   If analyses for other congeners are to
be conducted,  standards  will  have to be  custom synthesized and  characterized
as to identity  and purity.  Buser (1987)  has  demonstrated the microsyntheses
of several of  the  mixed  congeners by gamma  irradiation  of PCDDs  and  PCDFs in
dibromoethane.   While standards of known purity were not  produced, this tech-
nique may  prove useful  in the generation of  standards  for qualitative  use,
for  example,  in the  determination of  gas chromatographic retention times.

     E.   Quality Assurance/Quality Control (QA/QC)

          A strong QA/QC program  is necessary to support  the method guideline
approach which  has been  developed for  this  program.   To ensure  that  the data
obtained are of  known quality, the QA/QC program detailed  in Appendix A (Sec-
tion 15) includes provisions for  the use of carbon-13 labeled surrogates  where
possible.  In  addition,  replicate sample analysis, the routine analysis  of
spiked samples,  and  method blanks have been  incorporated.  Calibration pro-
cedures  for  instrumentation  are also clearly defined  for implementation  by
the analyst.


IV.  EXISTING METHODS

          A computer search, conducted on the CAS database.-.(1967-1984) yielded
no references concerning the analysis of HDDs and HDFs  in  commercial products.
However,  analytical  methods  for the  determination of  polychlorinated dioxins
(PCDDs), polychlorinated dibenzofurans (PCDFs), and polychlorinated and poly-
brominated biphenyls  (PCBs and PBBs)  in  various  matrices  have been  reported.
It  is  anticipated that  the analytical  procedures  for  PCDDs and  PCDFs,  summa-
rized below, may be modified  for  the analysis of bromo- and bromo/chloro-HDDs
and HDFs.

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             Table 2.   Currently Available HOD and HDF Standards
     Unlabeled
                                               Stable isotope labeled
Chlorinated dioxins:
Mono:

Di:


Tri:
        2,3
        2,8
        2,7/2,8e

        1,2,3
        1,2,4
        2,3,7

Tetra:   1,2,3,4
Penta:


Hexa:


Hepta:
Octa:
        1,2,3,7/1,2,3,8"
        1,2,3,7/1,2,8,9!
        1,2,4,7/1,2,4,8*
        1,2,6,7/1,2,8,9*
        1,2,7,8
        1,2,8,9
        1,3,6,8
        1,3,6,8/1,3,7,9*
        1,3,7,8
        2,3,7,8
        1,2,3,4',7
        1,2,3,7,8
        1,2,4,7,8

        1,2,3,4,7,8
        1,2,3,6,7,8
        1,2,3,7,8,9
        1,2,3,4,6,7,8
        1,2,3,4,6,7,8,9
Brominated dioxins:
Mono:

Oi:


Tri:

Tetra:
Penta:
        2,7
        2,8
        2,3,7

        1,2,3,4
        1,3,6,8/1,
        1,3,7,8
        2,3,7,8

        1,2,3,7,8
        1,2,4,7,8
                  3,7,9e
Hexa:   1,2,3,4,7,8
2,7/2,8 (u-13C12, 99%)£
                                               1,2,3,4 (13C6, 99%)
                                               2,3,7,8 (U-13C12> 99%)
                                               2,3,7,8 (U-37C14, 96%)C
                                               1,2,3,4 (U-13C12, 99%)
1,2,3,7,8 (U-13C12, 99%)


1,2,3,6,7,8 (U-13C12, 99%)
1,2,3,7,8,9 (U-13C12, 99%)


1,2,3,4,6,7,8 (Uri3C12, 99%)

1,2,3,4,6,7,8 (U-13C12, 99%)
        1,2,3,6,7,8/1,2,3,7,8,9'
2,3,7,8 (U-i3C12l 99%)




1,2,3,7,8 (U-13C12, 99%)

1,2,3,4,7,8 (U-i3C12, 99%)

-------
                             Table 2 (continued)
     Unlabeled                                 Stable isotope  labeled

Hepta:  1,2,3,4,6,7,8
Octa:   1,2,3,4,6,7,8,9
Mixed Br/Cl dioxins:
Tri:    7-9romo-2,3-dicnloro
Tetra:  2,3-Oi bromo-7,8-di ch1oro
        2,8-Oi bromo-3,7-di chloro
        2-8romo-3,7,8-di chloro
Penta:  2-Bromo-l,3,7,8-tetrachloro
Hexa:   3-3romo-l,2,4,7,8-pentachloro
Chlorinated dibenzofurans:
Mono:


01:




Tri:











Tetra:










2
3
4
2,3
2,6
2,7
2,8
4,6
1,2,3
1,2,4
1,3,6
1,3,7
1,6,7
2,3,4
2,3,6
2,3,8
2,4,6
2,4,7
2,4,8
2,6,7
1,2,3,4
1,2,3,6
1,2,3,7
1,2,3,8
1,2,3,9
1,2,4,6
1,2,4,8
1,2,4,9
1,2,7,3
1,3,4,6
1,3,4,7
                                               2,3,7,8 (U-i3C12, 99%)

-------
                             Table 2 (continued)
     Unlabeled
                                       Stable isotope labeled
Tetra:  1,3,4,8
(cont.) 1,3,4,9
        1,3,6,7
        1,3,6,8
        1,3,6,9
        1,3,7,8
        1,3,7,9
        1,4,6,7
        1,4,7,8
        2,3,4,6
        2,3,4,7
        2,3,4,8
        2,3,4,9
        2,3,6,7
        2,3,6,8
        2,3,7,8
        2,4,6,7
        2,4,6,8
        3,4,5,7
Penta:
Hexa:
1,2,3,4,6
1,2,3,4,7
1,2,3,4,8
1,2,3,4,9
1,2,3,6,7
1,2,3,7,8
1,2,3,7,9
1,2,3,8,9
1,2,4,6,7
1,2,4,7,8
1,3,4,6,7
1,3,4,7,8
2,3,4,6,7
2,3,4,6,8
2,3,4,6,9
2,3,4,7,8
2,3,4,7,9
1,2,
1,2,
1,2,
1,2,
1,2,
1.2,
1,2,
1,2.
1,3,
1,3,
2,3,
3,4,7,8
3,4,7,9
3,4,8,9
3,6,7,8
3,6,8,9
3,7,8,9
4,6,7,8
4,6,8,9
4,6,7,8
4,6,7,9
4,6,7,8
                                   1,2,3,7,8 (U-13C12, 99%)
1,2,3,4,7,8 (U-"C12, 99%)

-------
                             Table 2 (concluded)
     Unlabeled
                                               Stable isotope labeled
Hepta:   1,2,3,4,6,7,8
        1,2,3,4,6,8,9
        1,2,3,4,7,8,9
Octa:   1,2,3,4,6,7,8,9
Brominated dibenzofurans:
D1:
Tri:
Tetra:
Penta:
Hexa:
2,7
2,8
1,3,8
2,3,7,8
1,2,3,7,8
1,2,3,4,7,8
Mixed Br/CI furans:
Tri:    8-Bromo-2,3-dichloro
Tetra:  6,8-Oibromo-2,3-dichloro
        8-Bromo-2,3,4-trichloro
Penta:  6,8-Oibromo-2,3,4-trich1oro
                                       1,2,3,4,6,7,8 (U-«C12, 99%)
                                       1,2,3,4,6,7,8,9 (U-*3C12, 99%)
2,3,7,8 (U-13C12, 99%)
1,2,3,7,8 (U-13C12, 99%)
1,2,3,4,7,8 (U-13C12, 99%)
 aMixture  of  Indicated  isomers.   The  relative  amounts  are  unknown;  therefore,
  these  standards  are not useful  for  quantitation  calibration.
 bU-13C12  indicates  that the compound is  universally labeled with carbon-13.
 CU-37C14  indicates  that the compound is  universally labeled with chlorine-37.
                                        10

-------
     A.   Instrumental Methods

          Standard analytical  methods are  available for  the  analysis of
2,3,7,8-TCDD  in  soils  and sediments and water/wastewater  (EPA Method 613,
RCRA Method 8280,  other  methods under the  EPA Contract  Laboratory Program;
Harless et al.  1980;  McMillin et al.  1983).  These  methods  have  been vali-
dated for the measurement of 2,3,7,8-TCDD to detection levels of  1 ng/g (ppb)
for soils and sediments  and  1  ng/L  (ppt) for water/wastewater.  The  soil  and
sediment protocol  has  been  used for the analysis of over 11,000  samples  for
2,3,7,8-TCDD in EPA Region VII through the Contract Laboratory Program (CLP).
These methods have not been validated for  the analysis  of total PCDDs and
PCDFs.   However,  several  laboratories  have  provided  sufficient documentation
to suggest that  modification of  these  procedures  can  extend  the analysis  for
total tetrachloro- through octachloro-PCDDs and PCDFs.

          The extraction and cleanup procedures for the determination of  PCDDs
and PCDFs in  phenoxyalkanoic homicides  has been  reviewed  by  Baker  et al.
(1981).   Extraction  tecnniques  have  included steam distillation of TCDD  from
an alkaline solution of technical 2,4,5-T (Brenner et al. 1972, 1974), hexane
extraction from  an alkaline solution of 2,4,5-T,  liquid-liquid partitioning
of TCOD into hexane from a solution of technical  2,4,5-T in dimethylformamide/
acetonitrile/water (Vogel 1976), and separation of TCDD from isobutoxy methyl
esters of 2,4,5-T and fenoprop using a silica gel column (Ramsted et  al.   1977).

          Woolson et al.  (1972)  evaluated  17 different pesticides containing
the polychlorophenoxy group for the presence of PCDDs.  Extraction was accom-
plished using hexane extraction from methanolic potassium hydroxide  solution.
Cochrane et al.  (1981) described the  analysis of technical  and  formulated
products of 2,4-0 as the ester, free acid, and the amine.   The 2,4-D esters
were transferred  directly onto  a silica  column and eluted with 30% dichloro-
methane in hexane.  The 2,4-0 acids were dissolved in acetonitrile/water, and
the PCOOs were  partitioned  into hexane.   The 2,4-0 amines were dissolved in
water and partitioned into hexane.

          Analysis of  pentachlorophenol  (PCP)  for PCDOs  has  been  reported by
several researchers (Buser 1975, Buser and Bosshardt 1976,  Mieure et  al.   1977,
Blaser et al. 1976,  Firestone  et al.  1972,  Plimmer et al.  1973,  Lamberton et
al. 1979, Nelsson and  Renberg 1974, Pfeiffer et  al.  1978,  and Miles et  al.
1984).   The analytical  methods included dissolution  in  a  polar  solvent  and
partitioning  the  PCODs  into  a  nonpolar solvent,  separation  and collection of
PCDOs by  high performance  liquid chromatography  (Miles  et al. 1984), and
separation from the commercial product using a macro silica adsorption column
(Mieure et al.  1977).   Yamagishi et al.  (1981) reported a similar procedure
for the determination  of PCDDs  and  PCDFs in commercial diphenyl ether herbi-
cides such as nitrophen and chloromethoxynil.

     B.   Bioanalytical Methods

          A possible alternative to instrumental analysis for HDDs  and HDFs
may arise through the  further development of existing bioanalytical  or bio-
assay techniques.  These bioanalytical techniques  include (1) radioimmunoassay
(Albro et al.  1979,  McKinney et al. 1981), (2) aryl  hydrocarbon  hydroxylase
                                      11

-------
(AHH)  induction  assay (Bradlaw and Casterline  1979),  (3) cytosol  receptor
assay  (Hutzinger  1979),  (4)  early life stage  (E.L.S.)  bioassay (Helder and
Seinen 1985), and (5) in vitro keratinization assay (Knutson and  Poland  1980,
Gierthy et al.  1985).   Each of these  techniques  has  been evaluated  as  an
alternative technique for  screening  for the presence  of  PCDDs and/or PCDFs
based on biological/biochemical properties.

          The  radioimmunoassay,  AHH,   and  the  cytosol  receptor assay have
previously been  reviewed (NRCC 1981; NRCC  1984).   The  disadvantages of these
techniques in  general are  that they do not necessarily respond specifically
to PCDDs  and  PCDFs,  they respond to other compounds such as halogenated bi-
phenyls, azoxybenzenes,  and nonhalogenated polynuclear aromatic hydrocaroons,
and each  technique is less  sensitive than  available analytical  methods.   The
primary advantages of these techniques are lower  cost  and  rapidity of test
results.

          The in vitro keratinization and E.L.S. bioassays more recently have
provided additional options and possibly more specificity for determining  the
presence of PCODs and PCDFs in general.  Both techniques have been  demonstrated
to give roughly  comparable  results with HRGC/MS analysis of total  PCDDs and
PCDFs  in  a PCS fire  soot (Gierthy et al.  1985) and fly ash from  a  municipal
incinerator (Helder and Seinen 1985).-

          It is important to note that each of the bioassay techniques is  most
sensitive to the presence of 2,3,7,8-TCDD.   It is speculated that the  relative
response to other HDDs and HDFs might be dependent on halogen suostitution in
the 2,3,7,8-positions.   It  is  also important to note that the range  of com-
pounds evaluated-  with each  of  these bioassay techniques  is  limited.   Evalua-
tion of commercial products for  the presence  of  HDDs  and HDFs with  any of
these  bioassay  techniques  should  be completed using a  strong  QA/QC program
(Appendix A,  Section 15).  It is  suggested that the HDDs and HDFs be  isolated
from the commercial product matrix and other potential  contaminants using  the
methods specified  in  the guidelines (Appendix)  or  equivalent techniques.   If
bioassay techniques are used, it is advised that the analyst follow the recom-
mended QA/QC  program  (blanks,  spikes,  replicates,  etc.)  to  avoid  false nega-
tives.  It may also be necessary to use an alternate technique, such  as HRGC/MS,
to avoid false positives from the bioassay techniques.


V.  PROPOSED METHOD

          A flowchart for the general  guidelines for the analysis of  HDDs  and
HOFs  in commercial products  is shown in Figure 1.  This  scheme, adapted  from
general separation principles  and techniques reported in the literature,  is
based  on  the  physical/chemical  properties  of the commercial product;  there-
fore,  the exact extraction and cleanup procedure is dependent upon  the nature
of the product or formulation.

          The  properties utilized  in this  scheme  are volatility,  solubility,
acidity,  polarity, and  molecular  geometry.  For example,  if the  product is
sufficiently  volatile,  evaporation may be  employed.  Differential solubility
in immiscible  solvents  may  also be  the basis for  a liquid-liquid  extraction.
                                      12

-------
                 Comineiclul Pioducl
 PIIYSICAt/CIIEMICAL
 PROPERTIES
 SEPARATION
 fROM MATRIX
REMOVAL Of
INTERFERENCES
QUANIIIAIJON

Valollllly
Volatile
Nan-ValallU


• 1.2 Dichloiomelliane
• Bfomobeniene
•Vlnylchlorlda
Evaporation of
Sample lo Dryneil






Solubility In
Man-Polar Salveiili
Intolubla
• Titanium
• Plgmcnli
f xliocllon with
Nan-Polar .Solvent




Soluble

Dloxld.





Acidic
Acldlly
• Peiilubioaioulienul
•Malelc Acid
•UMiloioplienol
Batlc
Extraction
Bailc Alumina
Maciocoluiim





Neuhul



Nan
•2. 4.6- «an«
liibiomouiilllne
•2.4-Dlbtoiiia-4-
iilliounlllne
•3.4-Dlililuioanlllne
Extraction
Acidic Alumina
Maciocolunm

Mine-Column Clean Up
Alumina
Giuplilllied Caibon
HPLC Pliaiei









Planailly 1
• Oclabfoa»dlpheny|oiilde
Planar
• filbrouabe
Giaplilllzed Carbon of Melliylallan
SlllcolMaciocoluain Column Cliromalon.rapliy

Silica
Alumina
Floillll
HPLC
Graplilllzed Cuiban



'
Iniliumonlal Tetlinlquei
IIRGC/MS
IIRGC/ECI)


Blaaiiulylical Icclmiquai
Kerallnliallan Anay
RaJiolnmiuno Anuy
Allll Inducllon Ai.ay
Cyloiol-Heieplor Anuy
farly llfv Sluye Aiiuy
               Figure  1.  Tentative scheme for  the analysis  of  HDDs  and HDFs in  commercial  products.

-------
 For example, a salt may  be dissolved in aqueous  solution and the HDDs and
 HDFs extracted with an immiscible nonpolar solvent such as hexane.  The sol-
 vent,  number of extractions,  solvent-to-sample  ratio,  and other parameters
 are chosen at the  analyst's  discretion.   The planarity of the HDDs and HDFs
 may be exploited to separate  them from  a nonplanar matrix using graphitized
 carbon column  chromatography.   Column chromatography with  alumina,  silica, or
 ion exchange resins may also  be applicable  for use with some  matrices.   Other
 separation techniques  may also be  used if validated prior  to  sample analysis.
 Guidelines for the analysis   of HDDs  and HDFs in commercial  products  are
 presented  in Appendix  A.


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Norstrom A, Anderson K, Rappe C.  1976a.  Formation of chlorodibenzofurans  by
irradiation of chlorinated diphenyl ethers.  Chemosphere 5:21.

Norstrom  A,  Anderson  K,  Rappe C.   1976b.   Palladium  (II)  acetate promoted
cyclization  of  polychlorinated  diphenyl  ethers to  the  corresponding  dibenzo-
furans.  Chemosphere 5:419.

NRCC (National  Research  Council Canada).  1981.  Polychlorinated dibenzo-p_-
dioxins:  Limitations to  the  current  analytical  techniques.   NRCC No. 18576.

NRCC (National  Research  Council Canada).  1981.  Polychlorinated dibenzo-g-
dioxins:  Criteria  for  their effects on  man  and his environment.  NRCC No.
18574.

NRCC (National  Research  Council Canada).   1984.  Polychlorinated dibenzo-
furans:   Criteria for their  effects on  humans  and  the  environment.  NRCC No.
28846.
                                      17

-------
O'Keefe PW.  1978.   Format-ion  of brominated dibenzofurans from pyrolysis of
the polybnominated biphenyl  fire  retardant,  Firemaster  FF-1.   Environ Health
Perspect 23:347-350.

Pfeiffer CD, Nestn'ck  TJ,  Kocher CW.   1978 (May).   Determination of  chlori-
nated dibenzo-g-dioxins in purified pentachlorophenol by liquid chromatography.
Anal Chem 50(6).

Plimmer JR, Ruth JM,  Woolson EA.   1973.   Mass spectrometric identification of
the hepta-  and octa-chlorinated dibenzo-g-dioxins and dibenzofurans  in tech-
nical pentachlorophenol.   J Agr Food Chem 21(1).

Poland A, Glover  E.   1973.   Chlorinated dibenzo-p_-dioxins:  Potent inducers
of aminolevulinic acid synthetase and aryl hydrocarbon hydroxylase.   II.  A
study  of  the structure-activity  relationship.   Mol  Pharmacol  9:736-7^7.

Poland A,  Glover E.   1975.   Genetic expression of aryl hydrocarbon hydroxylase
by 2,3,7,8-tetrachlorodibenzo-g-dioxin.   Evidence for a receptor mutation in
genetically nonresponsive mice.  Mol Pharmacol 11:389-398.

Poland A, Glover  E,  Kende AS.  1976a.  Stereospecific,  high affinity  binding
of 2,3,7,8-tetrachlorodibenzo-g-dioxin by hepatic cytosol.   Evidence  that the
binding species is receptor for  induction of aryl  hydrocarbon  hydroxylase.
J Biol Chem 251:4936-4946.

Poland A, Glover  E,  Kende AS,  DeCamp  M, Giandomenico CM.   1976b.   3,4,3',4'-
Tetrachloroazoxybenzene and  azobenzene:  Potent  inducers of aryl hydrocarbon
hydroxylase.  Science 194:627-360.

Poland A, Greenlee WF, Kende AS.   1979.   Studies on the mechanism of  action
of the chlorinated dibenzo-g-dioxins  and related compounds.  Ann NY Acad Sci
30:214-230.

Polzhofer K.  1979.   Z Lebensm-Unters Forsch 168:21.

Ramstad T, Mahle NH,  Matalan R.  1977.  Anal Chem 49:386.

Rappe C, Buser HR, Bosshardt HP.   1978.   Chemosphere 7:431.

Rappe C, Gara A,  Buser HR.   1978a.  Identification of polychlorinated  dibenzo-
furans (PCDFs) in commercial chlorophenol  formulations.   Chemosphere  12:981-991.

RCRA Method 8280.  Method  of Analysis for chlorinated dibenzo-g-dioxins and
dibenzofurans.

Storherr RW, Watts RR, Gardner AM, Osgood T.  1971.   JAOAC 54:218.

Villanueva  EC, Jennings  RW, Burse V.  1974.  Evidence  of chlorodibenzo-g-
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916-917.

Vogel H, Weenen RO.   1976.   Fresenius Z Anal Chem 280:9.
                                      18

-------
Vos JG, Koeman JH, Van der Mass HL, Ten Noever de Brawn MC, de Vos RH.  1970.
Identification and toxicological  evaluation of chlorinated dibenzofuran and
chlorinated naphthalene  in  two commercial  polychlorinated biphenyls.   Food
Cosmet Toxicol 8:625-633.
                                •
Wang DKW,  Chiu OH, Leung PK,  Thomas  RS,  Lao RC.   1983.   Sampling  and analyt-
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facilities.  Environ Sci Res 26:113-126.

Williams IH.  1973 (November).   Photochemical  oxidation  of hexane isomers  to
compounds  sensitive  to  electron capture detection.  J Chrom Sci  11:593-596.

Woolson EA, Thomas RF, Ensor POJ.  1972.  Survey of polychloroaibenzo-£-dioxin
content in selected pesticides.  J Agr Food Chem 20(2):351-.

Yamagishi  T,  Miyazaki  T,  Akiyama  K,  Morita  M,  Nakagawa J,  Horii S,  Kaneko  S.
1981.  Polychlorinated  dibenzo-g-dioxins and dibenzofurans in  commercial  ai-
phenyl ether herbicides and in  freshwater fish collected from the application
area.  Chemosphere 10:1137-1144.

Yoshihara  S,  Nagata  K,  Yoshimura H, Kuroki  H,  Masuda Y.  1981.  Inductive
effect of  hepatic enzymes and  acute toxicity  of  individual  polychlorinated
dibenzofuran congeners in rats.  Toxicol Appl Pharmacol 59:580-588.
                                      19

-------
                         APPENDIX A
GUIDELINES FOR THE ANALYSIS OF HALOGENATED DIBENZQ-a-QIOXINS
          AND DIBENZOFURANS IN COMMERCIAL PRODUCTS
                             A-l

-------
        GUIDELINES FOR THE ANALYSIS OF HALOGENATED DIBENZO-p_-OIOXINS
                  AND OIBENZOFURANS IN COMMERCIAL PRODUCTS
1.0  Scope and Application

     1.1  These guidelines are applicable for the determination of halogenated
          dioxins (HDDs) and halogenated dibenzofurans (HOFs) in commercial
          products.

     1.2  These guidelines are restricted to use by or under the the super-
          vision of analysts experienced in the use of high resolution gas
          chromatograpny (HRGC) with mass spectrometric (MS) or electron
          capture detection (ECD).

     1.3  The limit of detection (LOO) and limit of quantitation (LOQ) are
          dependent upon the specific HDDs and/or HOFs monitored, the mass or"
          sample extracted, the complexity of the sample matrix, the sensi-
          tivity of the instrument, and the ability of the analyst to remove
          interferences and properly maintain the analytical systems.  The
          method accuracy and precision should be determined by each labora-
          tory for each commercial product.  Specific LOQs are given in the
          test rule (EPA 1985).

     1.4  The validity of the results depends on equivalent recovery of tne
          analytes and the surrogate compounds.  If the surrogates are not
          thoroughly incorporated into the matrix, the method is not applicable.

     1.5 'Certain analytical parameters and equipment designs, which will
          affect the validity of the analytical results, have been identified.
          Proper use of the method requires that such parameters or designs
          should be used as specified.  An experienced analyst may make
          modifications to parameters or equipment identified by the term
          "recommended."  Each time such modifications are made to the method,
          the analyst should repeat the procedures in Section 15.2.


2.0  Summary

     2.1  The product should be sampled such that the specimen collected for
          analysis is representative of the whole.  Statistically designed
          selection of the sampling position, time, or discrete product units
          should be employed.  The sample should be preserved to prevent HDD
          and HDF loss prior to analysis.  Customary inventory storage may be
          adequate for commercial products.  Guidelines for sample collection,
          handling, storage, etc., are provided in Appendices C and D.

     2.2  The sample is mechanically homogenized and subsampled as necessary
          The sample is then spiked with surrogate compounds, and the surro-
          gates thoroughly incorporated by further mechanical agitation.
                                     A-2

-------
     2.3  The  surrogate-spiked  sample  is extracted and  cleaned  up  at  the  dis-
         cretion of  the  analyst.   Possible extraction  techniques  include
         liquid-liquid partition,  column  chromatography,  etc.   The sample
         volume and  solvent  composition are  then adjusted as necessary  for
         final analysis.

     2.4  The  HDD and HDF content of the sample  extract is determined by  an
         appropriate analytical methodology.  Possibilities  include  high
         resolution  gas  chromatography/electron impact mass  spectrometry
         (HRGC/EIMS) operated  in the  selected ion monitoring mode (SIM),  high
         resolution  gas  chromatography with  electron capture detection  (HRGC/
         ECD), or  other  analytical procedures whicn have  been  validated  oy
         the  testing laboratory.

     2.5  For  analysis by HRGC/MS,  the HDDs and  HDFs are identified by com-
         parison of  their retention time  and mass spectral  abundance ratios
         or peak areas to those  in the calibration standard.   Identification
         by HRGC/ECO is  achieved by comparison  of retention  times.

     2.6  HDDs and  HDFs are quantitated against  the response  factors  for  a
         mixture of  HDD  and  HDF  congeners, using the response  of  the surro-
         gates to  correct for  loss of analytes  during  sample workup  and  to
         determine instrument  variability.

     2.7  HDDs and  HDFs identified  by  the  SIM technique may be  confirmed  by
         full mass scan  HRGC/EIMS, chemical  ionization mass  spectrometry
         (CIMS), high resolution mass spectrometry (HRMS),  retention on  alter-
         nate GC columns, or other techniques,  provided that the  sensitivity
         and  selectivity of  the  techniques are  demonstrated  to be adequate.

     2.8  The  total analysis  time  is dependent on the sample  matrix and  the
         extent of workup employed.   The  time required for a single  HRGC/SIM
         or HRGC/ECD analysis  of a sample is estimated as approximately  1 h.

     2.9  Appropriate quality control/quality assurance (QA/QC) procedures
         are  included to estimate  the quality of the results.   These QA/QC
         procedures  include  the  use of control  charts, and the analysis  of
         blanks, replicates, and  standard addition samples.  A QA/QC plan
         should be developed by  each  laboratory.  QA/QC results should  be
         reported  with the analytical results on a lot/batch basis.


3.0  Interferences

     3.1  Method  interferences  may  be  caused  by  contaminants  in solvents,
         reagents, glassware,  and  other  sample  processing hardware,  leading
         to discrete artifacts and/or elevated  baseline iff chromatograms.
         All  of  these materials  should be routinely demonstrated  to  be  free
         from the  interferences  by the analysis of laboratory  reagent blanks
         as described  in Section  16.4.   These interferences  are especially
         noted when using electron capture detection.
                                     A-3

-------
     3.1.1  Glassware should be scrupulously cleaned.  All glass should
            be cleaned as soon as possible after use by rinsing with the
            last solvent used.  This should be followed by detergent
            washing with hot water and rinses with tap water and reagent
            water.   The glassware should then be drained dry and heated
            in a muffle furnace at 400°C for 15 to 30 min.  Some ther-
            mally stable materials may not be eliminated by this treat-
            ment.   Solvent rinses with acetone and pesticide quality
            hexane may be substituted for the muffle furnace heating.
            National dioxin study procedures snould be used.   (Volumetric
            ware should not be heated in a muffle furnace.)  After it is
            dry and cool, clean glassware should be stored inverted in a
            contamination-free area or capped with aluminum foil to
            prevent accumulation of dust or other contaminants.

     3.1.2  The use of high purity reagents and solvents will help to
            minimize interference problems.  Purification of solvents by
            distillation in all-glass systems may be required.   All
            solvent lots should be checked for purity prior to use.

3.2  Interferences may also arise from contaminants that are coextracted
     with the HDDs and HDFs from the sample.   The extant of matrix
     interferences will vary considerably, depending on the nature and
     diversity of the samples.  Examples are provided below.

     3.2.1  Halogenated diphenyl ethers yield mass spectral fragment ions
            with exactly the same mass and number of halogen atoms as
            the corresponding HDFs.   Buser (1975) and Mieure et al.
            (1977) have demonstrated that basic alumina column chromatog-
            raphy will separata the chlorodiphenyl ethers from the PCDDs
            and PCDFs.  Likewise, Huckins et al.  (1978) have demonstrated
            that a graphitized charcoal column will remove the chloro-
            diphenyl ether interferences.

     3.2.2  The halobenzyl phenyl ethers may yield mass spectral inter-
            ferences to HDDs with selected ion monitoring (SIM)  tech-
            niques.  These compounds can also be removed from sample ex-
            tracts by cleanup on alumina or graphitized carbon columns.

     3.2.3  The bis(chlorophenoxy)methanes have gas chromatographic re-
            tention characteristics similar to PCDDs.  Interferences to
            analysis may arise when using electron capture detection;
            however, these compounds are distinguishable from PCDOs by
            mass spectrometry.

     3.2.4  The halogenated phenoxyphenols can dehydro'halogenate in the
            hot inlet of the gas chromatograph to form HDDs.   This has
            been reported for the chlorinated phenoxyphenols (Baker et al.
            1981) and for S-hydroxy-2,21,4,4',5,5'-hexabromobiphenyl
            (Gardner et al. 1979).  It is expected that analogous reac-
            tions will occur for the mixed bromo/chloro phenoxyphenols.
                                A-4

-------
       In addition, cyclization of diphenyl  ethers and hydroxy-
       biphenyls to form furans has been reported (Norstrom 1976a,
       1976b).   In order to avoid false positives, these species
       must be excluded from the final  extracts, or the extracts
       must be methylated to prevent ring closure during injection.

3.2.5  Interferences can be caused by the HDDs and HDFs themselves.
       This occurs due to the potentially large number of congeners
       (Tables 1 and 2) and to the ion  clusters produced by the
       isotopes of bromine and chlorine.   The masses and relative
       abundances of the ions in the molecular ion clusters of the
       HDDs and HDFs are presented in Tables 3 and 4.   Possible
       interferences from the molecular ion  clusters of the HDDs
       and HDFs are presented in Table  5.  As an example, a signal
       observed at m/z 460 can arise from tribromochlorocribenzo-
       dioxin,  octachlorodibenzodioxin, bromohexachlorodibenzofuran,
       or dibromotetrachlorodibenzofuran.
                           A-5

-------
       Table 1.   Theoretical Combinations of Bromo-, Chloro-,
                 and Bromo/Chloro Dibenzo-p_-dioxins


                Halogen      No.  of theoretical   No. of theoretical
Degree of     substitution       positional       2,3,7,8 substituted
halogenation




2









4













6




Br
0
1

0
1
2

0
1
2
3

0
1
'2
3
4

0
1
2
3
4
5

0
1
2
3
4
5
6

Cl
1
0

2
1
0

3
2
1
0

4
3
2
1
0

5
4
3
2
1
0

6
5
4
3
2
1
0

congeners
2
2
4
10
14
10
34
14
42
42
14
112
22
70
• 114
70
22
298
14
70
140
140
70
14
448
10
42
114
140
114
42
10
472
congeners
0
0
0
0
0
0
0
0
0
0
0
0
1
1
3
1
1
7
1
5
10
10
5
1
32
3
9
27
30
27
9
3
108
                                 A-6

-------
Table 1. (concluded)
Halogen
Degree of substitution
halogenation Br
0
1
2
I
5
6
7
0
1
2
3
8 4 .
5
6
7
8

Cl
7
6
5
4
3
2
-•
0
3
7
6
5
4
3
2
1
0

No. of theoretical No. of theoretical
positional 2,3,7,8 substituted
congeners congeners
2
14
42
70
70
42
14
2
256
1
2
10
14
22
14
10
2
1
76
Total 1,700
HDDs
1
7
21
35
35
21
7
1
128
1
2
10
14
22
14
10
2
1
76
Total 351
2,3,7,8 substi-
                            tuted HDDs
          A-7

-------
Table 2.   Theoretical Combinations of Bromo-, Chloro-,
            and Bromo/Chloro Dibenzofurans
Classes
No. of halogens Br
. 0
1 1

0
2 1
2

0
, 1
3 2
3

0
1
4 2
3
4

0
.1
5 2
5 3
4
5

0
1
2
6 3
4
5
6

Cl
1
0

2
1
0

3
2
1
0

4
3
2
1
0

5
4
3
2
1
0

6
5
4
3
2
1
0

No. of
positional
congeners
4
4
8
16
28
16
60
28
84
• 84
28
224
38
140
216
140
38
572
28
140
280
280
140
28
896
16
84
216
280
216
84
16
912
No. of 2,3,7,8
substituted
congeners
0
0
0
0
0
0
0
0
0
0
0
0
1
2
4
2
1
10
2
10
20
20
10
2
64
4
18
48
60
48
18
4
200
                             A-3

-------
Table 2.  (concluded)
Classes
No. of halogens Br
0
1
2
-, 3
/
1 4
5
6
7

0
1
2
3
8 4
5
6
7
8



Cl
7
6
5
4

3
2
1
0

3
7
6
5
4
3
2
1
0 •

Total
HDFs
No. of
positional
congeners
4
28
84
140

140
84
28
4
512
1
4
16
28
38
23
16
4
1
'136
3,320

No. of 2,3,7,8
substituted
congeners
2
17
40
69

69
40
17
2
256
1
4
16
28
38
28
15
4
1
136
Total 666
2,3,7,8 sub-
                           stituted
                           HDFs
          A-9

-------
3.   Chdi-aclei islic  Ion*  and I lie I r Heldlive liotope Abundances
        in llifc Molecular Ion Clusters ol  HIM)*
Uuyi ee u (
lldluyeiidltiiii
UTTBr« " " 0 1
0 164: 100 262: 100
165: 13.2 263: 13 2
166. 1 2 264. 98.9
265: 12.9
266: 1.2






•





1 218. 100 296: 76 0
219. 13 2 297: 10 0
220. 33.8 296: 100
221: 4 4 299: 13.1
300: 25 4
301: 3.3









1
2 252: 100 330: 60.9
253: 13 1 331: 8 0
254. 66 4 332. 100
255. 8 6 333 13.1
256: 11 4 334. 46 5
257. 1 4 335. 6 0
336. 6 9








2
340:
341:
342:
343.
344:
345.











3/4:
375.
376:
377.
378:
379.
360:
381.








408:
409
410:
411.
412:
413:
414.

416






50.8
6.7
100
13.1
49.8
6 5











43.6
5 7
100
13.1
70.7
9.2
14.4
1.6








38.2
5.0
100
13.1
90.4
11.8
32.8
4 2
4 3






3
418:
419.
420:
421:
422:
423:
424:
425:









452:
453:
454:
455:
456:
457:
458:
459:
460:
461:






486.
487:
488:
489:
490:
491:
492:
493:
494:
495:
496:




34 0
4 4
100
13.1
98 5
12.8
32.9
4.2









25.9
3.4
84.7
11.1
100
13.0
49.6
6 4
a s
1 0






20.3
2.6
73 0
9.5
100
13 0
64.4
8 4
19. J
2 4
2 2




4
496: 17.3
497: 2.3
498: 67.9
499: 8.9
500: 100
501: 13.0
502: 65.8
503: 8 6
504: 16 6
505: 2.1






.
530: 14.2
531: l.a
532: 60.2
533: 7.8
534. 100
535: 13.0
536: 80.6
537: 10 5
548: 31 1
549: 4 0
540: 4 6


.


564: 11.8
565: 1 5
566. 54 2
567: 7 1
568. 100
569: 13.0
570. 94 6
571: 12 3
572: 46 0
573. 6 2
574- 12 J
575. 1.6
576: 1.3


5
574.
575
576.
577:
578:
579:
560.
561:
562.
503:
564.
50b.





6U8:
609.
610.
611.
612:
613:
614:
615:
616:
617.
618:
619:
620:



642.
643:
644.
645.
646.
647
648.
640.
G'JO
CM
652
653.
6b4-


10 4
1.4
51 0
6 6
100
13.0
90 3
12 6
46 6
6 3
9 8
1 2





7 9
1 0
41 5
5 4
89 0
11 6
100
13.0
61.6
6 0
19 6
2 5
2.5



6 2
< 1
J4 2
4.4
79 5
10 4
100
13 0
73 0
9.b
30 8
4 (1
6 9


6
652: b.3
653: < 1
654: 31.2
655: 4.1
656: 76.4
657: 9.9
658: 100
659: 13 0
660: 73 8
661: 96
662: 29 3
663: 3.6
664: 5 0




686: 4 2
687: < 1
668: 26 3
689: 3 4
690: 69 3
691: 9 0
692: 100
693: 13.0
694: 85 2
695. 11 0
696: 42 7
697: 5.5
698: 11.6
699: 1 5
700: 1.3

720. 3.5
721: < 1
722: 22 6
723: 2.9
724: 6 4
725: 8 2
726: 100
727: 11 0
726: 96 1
729. 12 5
730: 57 5
731. 7 4
732: 20 8
733. 2 7
734. 4 1
7
730. 3 0
731. < 1
732: 20.8
733: 2 7
734: 61.2
735: 8 0
736. 100
737: 13.0
736: 98 2
739. 12. B
740: 58.1
741: 7 5
742. 19.2
743: 2.5
744: 2.8


764: 2.3
765. < 1
766. 16.6
767: 2 2
768: 51 9
769. 6.8
770. 91 7
771: 11.9
772. 100
773 13 0
774. 68.9
775. 89 1
776. 29.2
777: 3.8
778: 6.9
















a
808:
809:
810.
Bll:
812:
813:
614:
815:
816:
617.
816:
819:
020:
821:
822.
B23.
824:































1.5
< 1
12.1
1.6
41 6
5.4
81 6
10.6
100
13.0
78 6
10 2
38 8
5 0
11.0
1 4
1.4
































-------
                                                       Table 3  ((.oiiUnueil)
Degree of
lid loijBiia lion
cTi Br>   '      0
3














4














S












286:
267:
268:
289:
290.
291:
292:








320:
321:
322:
323:
324:
325:
326:
327:
326:






354:
355.
356.
357:
3b8:
359.
360.
361:
362.




100
13 1
99.1
12 9
33.1
4.3
3. B








75.9
10.0
100
13.0
49.7
6 4
11.1
1 4
1.0






60.8
6 0
100 '.
13 0
66 0
6 6
21 9
2 8
3 7




364:
365:
366:
367:
366:
369:
370:
371:
372:






396:
399:
400:
401:
402:
403:
404:
405:
406:






432:
433:
434:
435:
436:
437.
438:
439.
440:
441.
442.


50.8
6.7
100
13.1
66.0
6 6
18.4
2.3
1.9






43.6
5.7
100
13.1
84.6
11.0
34.2
4.4
6.8






37.2
4.9
97.4
12.7
100
13 0
52 8
6.8
15 3
2.0
2.4


442:
443:
444:
445.
446:
447.
446:
449.
41)0:
451.
452:




476:
477:
478:
479.
400.
481:
482:
483.
484:
485:
486:




510:
511:
512.
513-
514.
515.
516.
517.
518:
519.
520.
521.
522
31.0
4.1
91.4
11.9
100
13.0
50.6
6.5
12.1
1.5
1.1




23.9
3.1
78.2
10.2
100
13.0
64.1
8.3
22.1
2.8
4.0




19.0
2.5
66.5
6.9
100
13 0
77.1
10.0
34.3
44 1
8 9
1.1
1.3
520:
521:
522:
523:
524:
525:
526:
527:
528:
529:
530:




554:
555:
556:
557:
558:
559:
560:
561:
562:
563:
564:
565:
566:


588:
589:
590:
591:
592:
593:
594:
595:
596:
597:
596:
599:
600.
16.4
2.1
6.4
6.4
100
13 0
78 4
10.2
32.6
4.2
6.9




13.5
1.8
57.6
7.5
100
13.0
91.6
11.9
48.0
61.9
14.5
1 8
2.4


10.9
1.4
49 8
6 5
95.5
12.4
100
13 0
62 7
8.1
24 2
3 1
5 7
596:
599:
600:
601:
602:
603:
604:
605:
606:
607:
606:
609:
610:


632:
633:
634:
635:
636:
637:
636:
639:
640:
641:
642:
643:
644:
645:
646:













9.3
1.2
45.6
5.9
92.5
12.0
100
13.0
62.0
8 0
22.0
2.8
4 2


7.1
< 1
37.4
4.9
82 5
10.7
100
13.0
72.7
9 4
32.4
4.2
8.7
1.1
1.3













6/6:
677.
678:
679.
680:
601:
682:
603:
684:
685:
C06:
667.
686.
689.
690:




























4 9
< 1
28 7
3 7
71.9
9 4
100
13 0
83.9
10.9
43.4
5.6
13 5
1 7
2.3





























-------
lable 3  (concludoil)
Degree ol
ciref^
6 JUB:
389.
390:
391:
392:
393:
394:
395:
3U6:
197.
398:


7 422.
423:
424:
425.
426.
427:
428:
429:
430:
431.
432.


8 456:
457:
458:
459.
460:
461:
462:
463.
464:
465.
466:
467.
468.
0
50.8
6.6
100
13 0
82.2
10 7
36.2
4 7
9 0
1 1
1.2


43.6
5.7
100
13.0
98.6
12.8
54.1
7.0
17.9
2.3
3.6


33.2
4.3
87.1
11 3
100
13.0
65 8
8 5
27.1
3.3
7 2
< 1
1.2

466:
467:
468:
469:
470:
471:
472:
473:
474:
475:
476:


500.
501.
502:
503:
504:
505:
506:
S07:
508:
509:
510:
511:
512:






t
\





1
28.2
3 7
83.1
10 8
100
13 0
64.6
8.4
24.7
3.2
5.6


22 2
2.9
72 6
9.4
100
13 0
76.6
9.9
36.0
4.6
10.7
1 4
2.0














544:
545:
546.
547:
548.
549:
550:
551:
552:
553:
554.
555:
556.


























2345678
15.6
2 0
61.1
8.0
100
13 0
89.7
11 6
48 6
6.3
16.4
2.1
3 4



























-------
Table 4   Characteristic Ions and  [heir Relative  Isotope Abundances
              in the Molecular Ion Clusters of  111)1"s
Degree of
lldloijcnaliun
C'Tl Br. "0
0 168. 100 246:
169: 13 1 247:
248:
249.













1 202: 100 28Q:
203: 13 1 281.
204: 33 6 282.
205. 4 3 ' 283.
284:
285.








\
2 236: 100 314:
237: 13.1 315:
238: 66 2 316:
239. 8 6 317:
240- 11 3 318.
241: 1 4 319
320








1
100
13.1
98 7
12.9













76.1
10.0
100
13 0
25 2
3 2









61 0
8 0
100
13 0
46 4
6 0
6 8








2
324:
325:
326:
327:
328:
329-











358:
359:
360
361.
362.
363:
364:
365.







392
393.
394:
395.
396.
397
398
3'J'J
400






50.9
6.7
100
13 1
49.6
6 4











43.6
5.7
100
13.0
70.5
9.2
14.3
1 8







38.2
5 0
100
13.0
90.3
11.8
32 6
4.2
4 2






3
402:
403:
404:
405:
406:
407:
408:
409.









436:
437:
438:
439:
440:
441:
442:
443:
444:
445:





470:
471:
472:
473:
474:
475:
476:
477:
478.
479.
480:




34.0
4.4
100
13.0
98.4
12.8
32.7
4.2









25.9
3.4
84.8
11.0
100
13 0
49 4
6.4
8.4
1.0





20.3
2.6
73 0
9.5
100
13 0
64 3
8 3
19 2
2 4
2 2




4
480:
481:
482:
483:
484:
.485:
486:
487:
488:
489:







514:
515:
516:
517:
518:
519:
520:
521.
522:
523:
524:




548:
549:
550:
551:
552.
553.
554
555.
556.
557-
558.
559.
560.


17.3
2.3
67.9
a. a
100
13.0
65.7
8 5
16.4
2.1







14.2
1.8
60.2
7.8
100
13 0
80.5
10.4
31.0
4.0
4.5




11.8
1.5
54.2
7.0
100
13.0
94.5
12 2
47 9
6.2
12 2
1.6
1.3


5
55U:
559.
560.
561:
562.
563.
564.
565:
566:
567.
568.
569





592
593.
594.
595.
59G.
597.
598.
599.
600.
601:
602.
603:
604.


626.
627.
628:
629:
630.
631.
632
631
614.
Gib.
GJG.
637
GUI


10 4
1 4
51 0
6.6
100
13.0
98 2
12 7
48 5
6 2
9 a
1 2





8.0
1.0
41.6
5 4
89.1
11 6
100
13 0
61 5
8 0
5 4
2 5
2 5


6 2
< 1
34.2
4 4
79 6
10 3
100
13 0
7.1 0
'J 4
30 /
3'J 3
6 9


6 /
636:
637:
638:
639:
640:
641:
642:
643:
644:
645:
646:
647:
648.




670:
671:
672:
673:
674:
675:
676:
677:
£78:
679-
660:
681.
682:
683:
684:
704:
705:
706:
707:
708.
709.
710.
711
712.
713
714.
715-
716.
717
718
.5.3
< 1
31 2
4.1
76 5
19 9
100
13.0
73.7
9.5
29 2
3.7
4.9




4.2
< 1
26 4
3.4
69 3
9.0
100
13 0
65 1
11 0
42 6
5.5
11 5
1 5
1 3
3 5
< 1
22 6
2 9
63 6
8 2
100
13 0
96 0
12 4
57 4
7.4
20 /
2.G
4 1
714:
715:
716:
717.
718.
719.
720.
721:
722:
723:
724.
725.
72G:
727
720


748
749.
7bO.
/SI.
752.
753.
754.
755.
/5G.
757.
/58
759.
760.
761.
/G2.















3.0
< 1
20 8
2.7
61 2
7.9
100
12 9
98.1
12.7
57 9
7.5
19 1
2.4
2 8


2 3
< 1
1G 7
2 2
52 0
6 7
91 8
11 9
100
12.9
60 8
a 9
29 1
3 7
6 9















a
792:
793:
794.
795:
796:
797.
798.
799.
800.
001.
802:
803.
804:
805.
806:
807.
808:






























1.6
< 1
12.1
1.6
41.6
5.4
81 6
10.6
100
12 9
78 5
10.1
38 7
5.0
11 0
1 4
1 4































-------
                                                    Table 4  (continued)
Decree ot
lid logem II on
Clt Br> 0
3 270:
271:
272:
273.
274:
275:
276.








4 304:
305:
306.
307:
308.
309:
310:
311.







5 338.
339:
340:
341:
342:
343:
344.
345.
346.




100
13 1
08 9
12 9
32 9
4 2
3.8








76.0
9 9
100
13 0
49.5
6.4
11.0
1.4







60.9
8 0
100 •
13 0"
65 8
8 5
21 8
2.8
3 6




348.
349.
350:
351:
352:
353:
354.
355.
356:






382:
383:
384.
385:
386:
387.
388.
389.
390.






416:
417:
418:
419:
420:
421:
422:
423:
424.
425.
426:


1
SO. 9
6 6
100
13.0
65.9
8 5
18.3
2.3
1.9






43.6
5 7
100
13 0
84.5
11 0
34.1
4 4
6.7






37.2
4.8
97.5
12 7
100
13 0
52.7
6 8
15.3
1.9
2.3


2
426.
427.
428:
429.
430.
431:
432:
433:
434:
435:
436:




460:
461:
462.
463:
464:
465:
466.
467.
46B:
469:
470:




494:
495:
496:
497:
490.
499.
500:
501.
502:
503:
504.
50b:
SOG
31.1
4.0
91 5
11.9
100
13.0
50.5
6 5
12.1
1 5
1.1




23.9
3.1
78.3
10.2
100
13.0
64 0
8 3
22.0
2.8
3.9




19.1
2.5
68.6
8 9
100
13.0
77.0
10.0
34.1
4.4
8.8
1 1
1.2
3
504:
505:
506:
507:
508:
509:
510:
511:
512:
513:
514:




538:
539:
540:
541:
542:
543:
544:
545:
546:
547:
548:
549:
550:


572:
573:
574:
575:
576:
577:
578:
579.
580:
581:
582:
583:
584:
16.4
2.1
64. J
8 4
100
13.0
78.3
10.1
32.4
4.2
6.8




13.6
1.8
57.6
7.5
100
13.0
91 6
11.8
47.9
6.1
14.4
1.8
2.3


10.9
1.4
49.9
6.5
95 6
12.4
100
12.9
62 6
8.1
24 2
3 1
5.6
4
582:
583:
584:
585:
586:
587:
588.
589:
590:
591:
592:
593:
594:


616:
617:
618:
619:
620:
621:
622:
623:
624:
625:
626:
627:
628:
629:
630:













9.3
< 1
45.7
5.9
92.5
12 0
100
13.0
61.9
8 0
21.9
2 8
4.1


7.2
< 1
37.4
4.9
8 2
10.7
100
12.9
72.6
9.3
32.3
4.1
8.7
1.1
12.9













567
660:
661.
662:
663.
664:
665.
6G6:
667.
668.
669:
670:
671:
672:
673.
674:




























4 9
< 1
2H.O
3 7
72 0
9 3
100
12.9
83 8
10 8
43.3
5.6
13 4
17.0
2 3





























-------
Table 4.  (concluded)
Ouijrce at
lldlouenalluii
crre?* 	
6 372.
373:
374.
375:
376:
377.
378.
379.
380
381:
382:


7 406:
407:
408:
409.
410:
411:
412.
413.
414.
415.
416:


8 440.
441:
442:
443.
444:
445.
446:
447.
448:
449.
450:
451:
452:
0
50 8
6 6
100
13 0
82.1
10 6
36.1
4 6
9 0
1.1
1 2


43.6
5.7
100
13.0
98.4
12.7
53 9
7.0
17.8
2 2
3 6


33 3
4 3
87.2
11.3
100
12 9
65 7
B 5
27 0
3 4
7 1
< 1
1 2

450:
451:
452:
453:
454.
455:
456:
457:
458:
459:
460.


484:
485:
486:
487.
488:
489:
490.
491:
492:
493:
494:
495:
496:






*






1
2 8
3.7
83.2
10 6
100
12 9
64 7
8.4
24 6
3 1
5 6


22.2
2.9
72.7
9.4
100
12.9
76 5
9 9
36.0
4.6
10.7
1.3
2 0














528:
529.
530:
531.
532:
533.
534.
535.
536.
537:
538:
539:
540.


























2345678
15.6
2.0
61.1
7.9
100
12 9
89 6
11 6
48 4
6.2
16.3
2 1
3.4



























-------
Table 5.   Potential Interferences from Complex Mixtures
                    of HDDs and HDFs
_/-
ffl/z
163
169

184
185
136
202
203
204
205
218
219
220
221
236
237
233
239
240
241
246
247
243
249
252
253
254
255
256
257
262
263
264
265
266
270
271
272
273
274
275
275
230
281
282
283
284
285


Fa
F.
n
0D
0
0
Cl-F
Cl-F
Cl-F
Cl-F
Cl-0
Cl-0
Cl-D
Cl-0
CT2-F
C12-F
C12-F
C12-F
C12-F
C12-F
Br-F
Br-F
Br-F
Br-F
C12-0
C12-0
C12-0
C12-0
C12-0
C12-0
Br2-D
Br2-0
Br2-0
Br2-0
Br2-0
C13-F
C13-F
C13-F
C13-F
C13-F
C13-F
C13-F
ClBr-F
ClBr-F
C18r-F
ClBr-F
ClBr-F
ClBr-F
                             A-16

-------
Table 5.  (continued)
/
m/z
286
287
288
289
290
291
292
296
297
298
299
300
301
304
305
306
307
308
309
310
311
314
315
316
317
318
319
320
321
322
323
324
325
326
327
328
329
330
331
332
333
334
335
336
338
339


C13-0
Cl,-0
C13-0
C13-D
C13-D
C13-0
C13-0
ClBr-0
CIBr-0
ClBr-D
CIBr-Q
CIBr-0
CIBr-0
C14-F
C14-F
C14-F
C14-F
C14-F
C14-F
C14-F
C14-F
Cl2Br-F
Cl2Br-F
C12Br-F
C12Br-F
Cl2Br-F
Cl2Br-F
Cl2Br-F C14-0
C14-0
C14-0
C14-0
C14-0 Br2-F
C14-D Br2-F
C14-0 Br2-F
C14-0 Br2-F
C14-0 Br2-F
Br2-F
Cl2Br-0
Cl2Br-D
Cl,Br-0
Cl2Br-0
Cl2Br-0
Cl2Br-0
Cl,Br-0
C15-F
C15-F
          A-17

-------
Table 5. (continued)
/
m/z
340
341
342
343
344
345
346
348
349
350
351
352
353
354
355
356
357
358
359
360
361
362
363
364
365
366
367
368
369
370
371
372
373
374
375
376
377
378
379
380
381
382
383
384
385
386


C1S-F
Clg-F
Cl.-F
CVF
C15-F
C1S-F
C15-F
Cl38r-F
Cl3Br-F
Cl3Br-F
Cl38r-F
Cl3Br-F
Cl3Br-F
Cl3Br-F
Cl3Br-F
Cl3Br-F
C15-0
ci5-o
Clg-O
ci5-o
Cls-0
C15-D
ClBr2-F
ClBr2-F
ClBr2-F
Cl3Br-D
Cl3Br-D
Cl3Br-D
Cl3Br-0
Cl3Br-0
Cl3Br-0
Cl3Br-0
C16-F
C16-F
C1S-F
C16-F
C16-F
C16-F
C16-F
C1S-F
C16-F
C16-F
Cl48r-F
Cl4Br-F
Cl4Br-F
C14Br-F


Br2-0
Br2-0
8r2-0
Br2-Q
3r2-0
Br2-0







Cls-0
ci5-o
C15-0

ClBr2-F
C1Br2-F
ClBr2-F
C1Br2-F
C1Br2-F

Cl33r-0
C13Br-0






C1S-F

C1Br2-0
C1Br2-0
C1Br2-0
ClBr2-0
ClBr2-D
C1Br2-D
ClBr2-D
ClBr2-D
C14Br-F




          A-18

-------
Table 5. (continued)
-. /_
m/z
387
388
389
390
391
392
393
394
395
396
397
398
399
400
401
402
403
404
405
406
407
408
409
410
411
412
413
414
415
416
417
418
419
420
421
422
423
424
425
426
427
428
429
430
431
432


Cl48r-F
Cl4Br-F
Cl4Br-F
Cl48r-F
CVD
cvo
ci$-o
cvo
cvo
cvo
CVD
cvo
Cl2Br2-F
Cl28r2-F
C14Br-0
C14Br-0
Cl48r-0
Cl4Br-0
Cl4Br-D '
C14Br-0
Br3-F
Br3-F
Br3-F
C17-F
C17-F
C17-F
C17-F
C17-F
C17-F
C17-F
ClsBr-F
Cl5Br-F
Cl5Br-F
ClsBr-F
Cl5Br-F
Cls8r-F
Cl5Br-F
ClsBr-F
Cl5Br-F
ClsBr-F
C17-0
C17-0
C17-0
C17-0
C17-0
C17-0



CVD
CVO
CVO

Cl28r2-F
Cl28r2-F
Cl28r2-F
Cl2Br2-F
Cl2Br2-F
Cl,8r2-F
C12Br2-F Cl48r3-D
Cl4Br-0
Cl48r-0

Br3-F
Br3-F
Br3-F
' Br3-F
Br3-F C17.-F
• C17-F
C17-F C12Br2-0
C17-F C12Br2-D
Cl28r2-D
C12Br2-0
CT28r2-0
CT2Br2-0
C1,Br2-0
CT28r2-0
C12Br2-0 ClsBr-F
•
8r3-0
Br3-0
Br3-0
Br3-0
Br3-D C17-0
Br3-0 C17-0
8r3-0 C17-0
Br3-D C17-0
Br3-0 C17-0 " CT3Br2-
CT3Br2-F
CT3Br2-F
CT3Br2-F
Cl3Br2-F
Cl3Br2-F
Cl3Br2-F C158r-D
          A-19

-------
Table 5. (continued)
_ /_
m/z
433
434
435
436
437
438
439
440
441
4*2
443
444
445
446
447
448
449
450
451
452
453
454
455
456
457
458
459
460
461
462
463
464
465
466
467
468
469
470
471
472
473
474
475
476
477
478


Cl5Br-D
Cl5Br-D
Cl5Br-0
Cl3Br-0
Cl58r-0
ClsBr-D
ClsBr-0
Cl5Br-D
Cl5Br-D
Cl5Br-D
C18r3-F
C1Br3-F
ClBr3-F
Cl.-F
C18-F
C18-F
C18-F
CU-F
C18-F
C18-F
Cl6Br-F
Cl6Br-F
ClsBr-F
C16Br-F
Cl6Br-F
- ClsBr-F
Cl6Br-F
Cl68r-F
ClBr3-0
C18-0
ci8-o
C18-0
C18-0
C18-0
C18-0
C18-0
Cl4Br2-F
Cl4Br2-F
Cl68r-0
Cl6Br-0
ClsBr-0
Cl6Br-D
Cl6Br-0
Cl6Br-D
Cl4Br2-Q
Cl4Br,-0


C13BP2-F
Cl3Br2-F
C138r2-F
ClBr3-F
ClBr3-F
ClBr3-F
C18r3-F
ClBr3-F
C18r3-F
C18r3-F
C18-F
C18-F
C18-F
C13Br2-0
Cl3Br2-0
Cl38r2-0
CT3Br,-0
Cl,8r;-0
C138r2-0
C13Br2-D
C1Br3-0
C18r3-0
ClBr3-D
C18r3-0
C1Br3-0
ClBr3-0
ClBr3-0
ClBr3-0
C18-0
C14Br2-F
Cl4Br2-F
C14Br,-F
Cl4Br2-F
Cl4Br2-F
Cl4Br2-F
Cl4Br2-F
Cls8r-0
ClsBr-D
C128r3-F
C12Br3-F
Cl2Br3-F
C12Br3-F
Cl2Br3-F
Cl4Br,-0
Cl28r3-F
C128r3-F





C138r,-F



CVF
C13-F
C18-F
C138r2-0
Cl3Br,-0
C"l3Br2-0




C163r-F
Cl6Br-F
C16Br-F



C18-0
C18-0
C18-0
C18-0
C18-0

.



Cl6Br-0
C1sBr-0
Cl6Br-0

C12Br3-F





Cl23r3-F













C13Br2-0


•






ClBr3-0







Cl4Br2-F
Cl48r2-F

















         A-20

-------
Table 5.  (continued)
_ /_
m/z
479
480
481
482
483
484
485
486
487
488
489
490
491
492
493
494
495
496
497
498
499
500 .
501
502
503
504
505
506
507
508
509
510
511
512
513
514
515
516
517
518
519
520
521
522
523
524



Cl4Br2-0
Cl4Br,-0
C148r2-0
C14Brv>-0
Cl4Br^-0
C148r2-0
Cl4Bro-0
Cl4Br2-0
Br4-F
3r4-F
Br4-F
C17Br-F
Cl78r-F
Cl7Br-F
C178r-F
C178r-F
Cl2Br3-0
Cl28r3-0
Br4-0
Br4-0
Br4-0
Br4-0
Br4-0
Br4-0
Br4-0
Br4-0
Br4-0
C1sBr2-F
Cl5Br-0
Cl7Br-D
Cl78r-0
Cl7Br-0
C17Br-0
Cl7Br-0
Cl3Br3-F
Cl3Br3-F
ClsBr2-0
ClsBr2-D
Cl5Br2-D
ClsBr2-0
ClsBr2-D
Cl5Br2-0
C15Br,-0
Cl33r2-0
Br4-F
Br4-F



Cl28r3-F
8r4-F
Br4-F
8r4-F
Br4-F
Br4-F
Br4-F
8r4-F
C178r-F
CT78r-F
Cl78r-F
Cl28r3-0
C12Br3-0
CT28r3-0
CT2Br3-0
Cl28r3-0
C158r,-F
3r4-0"
CT5Br,-F
Cl5Br2-F
Cl58r2-F
Cl5Br2-F
C15Br2-F
ClsBr2-F
C15Br2-F
Cl5Br2-F
ClsBr2-F
Cl78r-0
C13Br3-F
C13Br3-F
Cl3Br3-F
CT38r3-F
CT38r3-F
Cl3Br3-F
Cl5Br2-0
ClsBr2-0
Br4-F
Br4-F
Br4-F
Br4-F
Br4-F
Br4-F
Br4-F
Br4-F
Cl3Br3-0
Cl3Br3-0
A-21

present ------

C12Br3-F



Cl73r-F
Cl73r-F
Cl78r-F
Cl,3r3-0
CU3r,-D
C12Br3-0




Cl58r,-F

Cl38r-F



Cl7Br-0
Cl7Br-0
C17Br-0
Cl7Br-D
C17Br-0
C17Br-0
C13Br3-F



CT5Br2-D
C13Br2-D
Cl3Br2-D

ClBr4-F





C13Br3-D
Cl3Br3-0
Cl3Br3-D












Ci,8r3-0

















CT3Br3-F
CT3Br3-F





















-------
Table 5. (continued)
m/z
525
526
527
528
529
530
531
532
533
534
535
536
537
538
539
540
541
542
543
544
545
546
547
548
549
550
551
552
553
554
555
556
557
558
559
560
561
562
563
564
565
566
557
563
569


Cl38r3-0
Cl3Br3-Q
Cl3Br3-D
Cl3Br3-0
CT3Br3-D
Cl38r3-0
C16Br2-F
Cl68r2-F
C16Br2-F
ClsBr2-F
C1sBr2-F
Cl68r2-F
Cl6Br2-F
Cl6Br2-F
ClsBr2-F
ClsBr2-F
C14Br3-F
C14Br3-F
Cl48r3-F
C14Br3-F
Cl4Br3-F
Cl48r3-F
Cl4Br3-F
Cl4Br3-F
Cl4Br3-F
C14Br3-F
Cl6Br2-0
ClsBr2-0
Cl6Br2-0
Cl6Br2-0
C16Br2-D
ClsBr2-0
Cl28r4-F
C128r4-F
C12Br4-F
Cl2Br4-F
Cl4Br3-0
C14Br3-0
Cl48r3-0
Cl4Br3-D
Cl4Br3-0
Cl4Br3-0
Br3-F
Brs-F
Br3-F



C16Br2-F
Cl6Br,-F
Cl6Br2-F
ClBr4-0
ClBr4-0
C1Br4-Q
ClBr4-0
C1Br4-0
C1Br4-0
C1Br4-0
ClBr4-0
ClBr4-0
ClBr4-0-



C16Br2-0 .
C16Br2-D
Cl6Br2-0
Cl5Br,-0
Cl6Br2-0
Cl68r2-D
Cl6Br2-0
Cl2Br4-F
Cl2Br4-F
C12Br4-F
Cl2Br4-F
Cl2Br4-F
Cl2Br4-F
C148r3-0
Cl4Br3-0
Cl48r3-0
Cl4Br3-0
Brs-F
Brs-F
Brs-F
Brs-F
Brs-F
Brs-F
Cl2Br4-0
Cl28r4-0
Cl2Br4-D





Cl3Br4-0







Cl48r3-F
Cl4Br3-F "
Cl4Br3-F






Cl2Br4-F
Cl28r4-F
Cl28r4-F
Cl28r4-F



C14Br3-0
Cl4Br3-0
Cl4Br3-0

Br3-F
8r5-F
Brs-F


Cl2Br4-0
Cl28r4-0
Cl2Br4-0
Cl28r4-0



          A-22

-------
Table 5.  (continued)
/_
m/z
570
571
572
573
574
575
576
577
578
579
530
581
582
583
584
585
586
587
588
589
590
591
592
593
594
595
596
597
598
599
600
601
602
603
604
605
606
607
608
609
610
611
612
613
614
615


Cl2Br4-D
Cl2Br4-D
Cl28r4-D
Cl2Br4-0
Cl2Br4-D
C128r4-D
C128r4-0
3r5-0
8r5-0
Brs-0
Br5-0
Brs-D
Brs-0
Brs-0
Brs-0
Br3-0
Cl3Br4-F
C138r4-F
Cl38r4-F
Cl3Br4-F
Cl3Br4-F
Cl38r4-F
Cl3Br4-F
C138r4-F
Cl3Br4-F
Cl5Br3-0
Cl5Br3-0
ClsBr3-D
Cl58r3-0
ClsBr3-D
ClsBr3-D
C1Brs-F
C1Brs-F
ClBrs-F
ClBr5-F
Cl38r4-0
CT3Br4-0
Cl3Br4-0
Cl3Br4-0
Cl3Br4-0
Cl3Br4-0
ClBr5-D
C18r5-0
ClBr5-0
ClBr3-D
C1Br3-0




Cl5Br3-F
Cl5Br3-F
Cl58r3-F
C15Br3-F
Cl5Br3-F
C158r3-F
C15Br3-F
C15Br3-F
Cl5Br3-F
Cl5Br3-F
Cl5Br3-F
Cl5Br3-F
CT5Br3-F
Cl38r4-F


C15Br3-0
Cl5Br3-0
ClsBr3-0
ClsBr3-0
Cl5Br3-0
C153r3-0
Cl5Br3-0
ClBr5-F
C1Brs-F
ClBrg-F
C18rs-F
ClBrs-F
ClBr5-F
C138r4-0
Cl38r4-D
Cl3Br4-0
C13Br4-0



C1Brs-0
ClBrs-0
ClBrs-D











3r5-0
Brs-0
3r5-0





CT38r4-F
Cl38r4-F
Cl3Br4-F







C1Brs-F
ClBr5-F
ClBrs-F



C13Br4-0
C13Br4-0
Cl38r4-0















          A-23

-------

m/z
515
517
618
519
520
621
622
523
524
525
625
627
628
629
630
631
632
533
634
635
636
637
638
639
640
641
642
643
644
645
646
647
64S
649
650
651
652
653
654
655
656
657
658
659
Table
5. (continued)



C1Brs-0
C18rs-0
ClBrs-0
C1Br5-0
C18r3-0
C148r4-F
Cl48r4-F
C14Br4-F
Cl4Br4-F
Cl4Br4-F
Cl4Br4-F
CT4Br4-F
CT4Br4-F
CT48r4-F
Cl48r4-F
Cl28r5-F
Cl28r5-F
Cl28rs-F
Cl28r5-F
CT28r5-F
Cl2Brs-F
Cl28r5-F
Cl2Brs-F
C148r4-0
Cl48r4-Q
Cl4Br4-D
Cl48r4-D
C148r4-D
Cl48r4-0
C148r4-D
Cl48r4-0
Brs-F
8r6-F
Cl28r5-0
Cl2Brs-0
C12Br5-0
Cl2Brs-0
Cl2Brs-D
C12Brs-0
Br6-D
Br6-D
Brs-0
8rs-0
8r6-0
Cl4Br4-F
Cl48r4-F
Cl4Br4-F
Cl4Br4-f
Cl43r4-F





C12Brs-F
C12Brs-F
Cl28r5-F
Cl28rs-F
Cl28r5-F

C148r4-0
Cl43r4-0
Cl4Br4-0
Cl4Br4-0
C14Br4-0
Cl4Br4-0
Cl48r4-0
Br6-F
Brs-F
Br6-F
Brs-F
8rs-F
Brs-F
Brs-F
Brs-F
C12BP5-0
Cl28rs-0



Brs-D •
Brs-D
Brs-D





•



















Brs-F '
Br6-F
Brs-F



Cl28rs-0
Cl2Brs-0
C12Br3-D
Cl28rs-D
Cl2Br5-0













A-24

-------
Table 5.  (continued)
_ /•»
m/z
660
661
662
663
664
665
666
667
668
669
670
671
672
673
674
675
676
577
678
679
680
681
682
683
684
685
686
687
688
689
690
691
692
693
694
695
696
697
698
699
700
704
705
706
707
708


Br6-0
Br6-0
Brs-0
3rs-0
Brs-0
Cl38r3-r
Cl38r5-F
Cl38r3-F
Cl3Br5-F
Cl3Br3-F
Cl38r5-F
Cl3Br5-F
Cl38r5-F
Cl3Brs-F
Cl38rs-F
ClBrs-F
ClBrs-F
ClBr6-F
ClBrs-F
ClBrs-F
ClBrs-F
ClBrs-F
C1Br6-F
ClBrs-F
ClBrs-F
Cl3Br5-0
Cl38rs-D
C138rs-0
C13Brs-0
Cl38rs-0
CT38r3-D
ClBrs-0
ClBrs-0
ClBrs-0
ClBrs-0
ClBrs-0
ClBrs-0
ClBrs-0
ClBr6-D
ClBrs-0
ClBr6-0
Cl2Br6-F
Cl28rs-F
Cl28r6-F
C12Brs-F
Cl2Br6-F

----- Ions present 	
Cl3Br5-F
Cl38r5-F
C13Srs-F
Cl3Br5-F
C13Brs-F





C1Brs-F
C1Brs-F
ClBrs-F
ClBr6-F
C1Br6-F

Cl3Br5-0
C138r3-0
Cl3Brs-0
C!3Brs-0
Cl3Brs-D
Cl3Br5-0
Cl3Brs-0
C138rs-0
Cl38r5-D -

C1Brs-0
ClBrs-0
C18r6-0
C18r6-D
ClBr6-0















         A-25

-------
Table 5. (continued)

m/z
709
710
711
712
713
714
715
716
717
718
719
720
721
722
723
724
725
726
727
728
729
730
731
732
733
734
735
736
737
738
739
740
741
742
743
744
748
749
750
751
752
753
754
755
756
757


Cl2Brs-F
CT2Br6-F
•Cl2Brs-F
Cl2Brs-F
Cl2Br6-F
Cl2Brs-F
C12Br6-F
Cl28rs-F
C12Brs-F
CT2Br6-F
3r7-F
Br7-F
Br7-F
Br7-F
Br7-F
Br7-F
3r7-F
Br7-F
8r7-F
Br7-F
Cl2Brs-0
Cl2Brs-0
Cl2Brs-0
Cl2Brs-0
Cl2Br6-0
C12Br6-0
Br7-D
Br7-0
Br7-0
Br7-0
Br7-0
Br7-0
Br7-0
Br7-D
Br7-0
Br7-0
ClBr7-F
C18r7-F
ClBr7-F
C1Br7-F
C1Br7-F
C18r7-F
C18r7-F
ClBr7-F
C18r7-F
ClBr7-F







Br7-F
Br7-F
3r7-F
8r7-F
8r7-F

Cl28rs-0
C128rs-0
C12Br6-0
Cl23rs-0
C128rs-Q
Cl28r5-D
Cl28rs-0
Cl2Brs-0
C12Brs-0

Br7-0
Br7-0
Br7-0
Br7-D
8r7-0




















         A-26

-------
Table 5. (continued)
/_
m/z
758
759
760
761
762
764
765
756
767
768
769
770
771
772
773
774
775
776
111
778
792
793
794
795
796
797
798
799
800
801
802
803
804
805
806
807
808
809
810
811
812
813
314
815
816


ClBr7-F
C1Br7-F
ClBr7-F
ClBr7-F
ClBr7-F
C1Br7-0
ClBr7-0
ClBr7-0
ClBr7-0
C18r7-0
ClBr7-0
C18r7-0
ClBr7-0
ClBr7-0
ClBr7-0
C18r7-0
ClBr7-0
C18r7-0
C18r7-0
ClBr7-0
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F
Br8-F Br8-0
Br8-0
Br8-0
Br8-D
Br8-0
Br8-0
Br8-0
Br8-0
Br8-D
         A-27

-------
                  Table 5. (concluded)
m/z          	  Ions present
817          Br8-0
818          Br8-D
819          Br8-0
320          8r8-0
821          8r8-D
822          Br8-0
823          Br8-0
324          Br3-0
 F = dibenzofuran.
 D = dibenzo-£-dioxin.
                           A-28

-------
4.0
     Some of the following safety practices are excerpted directly from the
     EPA Method 613, "Determination of 2,3,7,8-TCDD in Water and Wastewater,"
     Section 4 (October 1984 version).  Although these practices were designed
     for the safe handling of 2,3,7,8-tetrachlorodibenzo-g-dioxin (2,3,7,8-
     TCDD), they are also applicable for other HDDs and HDFs.

     4.1  2,3,7,8-TCDD has been tentatively classified as a known or suspected
          human or mammalian carcinogen.

          Note:  Congeners substituted in the 2,3,7,3 positions are reoorted
          to be the most toxic.  Extreme caution should be taken when handling
          these compounds as dry powders or in concentrated solutions.

          The toxicity or carcinogenicity of other reagents used in these
          guidelines has not been precisely defined; however, each cnenncal
          compound should be treated as a potential health hazard.   From this
          viewpoint, exposure to these chemicals must be reduced to the low-
          est possible level by whatever means available.  The laboratory  is
          responsible for maintaining a current awareness file of OSHA regu-
          lations regarding the safe handling of the chemicals specified in
          these guidelines.  A reference file of material data handling sheets
          should also be made available to all personnel involved in the
          chemical analysis.

     4.2  If diethyl ether is used, it should be monitored regularly to deter-
          mine its peroxide content.  Under no circumstances should diethyl
          ether be used with a peroxide content in excess of 50 ppm, as an
          explosion could occur.  Peroxide test strips manufactured by EM
          Laboratories (available from Scientific Products Company, Cat.  No
          P1126-8, and other suppliers) are recommended for this test.   Pro-
          cedures for removal of peroxides from diethyl ether are included in
          the instructions supplied with the test kit.

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

          4.3.1   Contamination of the laboratory will be minimized by
                  conducting all manipulations in a hood.

          4.3.2   The effluents of sample splitters for the gas chromatograph
                  and roughing pumps on the GC/MS should pass through either
                  a column of activated charcoal or be bubbled through a trap
                  containing oil or high-boiling alcohols.

          4.3.3   Liquid waste should be dissolved in methanol or ethanol and
                  irradiated with ultraviolet light with a wavelength greater
                  than 290 nm for several days.   (Use F 40 BL lamps or equiva-
                  lent.)  Analyze liquid wastes and dispose of the solutions
                  when 2,3,7,8-TCDD can no longer be detected.
                                    A-29

-------
4.4  Dow Chemical U.S.A.  has issued the following precautions (revised
     November 1978) for the safe handling of 2,3,7,8-TCDO in the labora-
     tory:

     4.4.1   The following statements on safe handling are as complete
             as possible on the basis of available toxicological infor-
             mation.   The precautions for safe handling and use are
             necessarily general in nature since detailed, specific
             recommendations can be made only for the particular expo-
             sure and circumstances of each individual use.   Inquiries
             about specific operations or uses may be addressed to the
             Oow Chemical Company.   Assistance in evaluating the health
             hazards of particular plant conditions may be obtained from
             certain consulting laboratories and from state Departments
             of Health or of Labor, many of which have an industrial
             health service.  2,3,7,8-TCDO is extremely toxic to labora-
             tory animals.  However, it has been handled for years with-
             out known injury in analytical and biological laboratories.
             Techniques used in handling radioactive and infectious ma-
             terials are applicable to 2,3,7,8-TCDO.

             4.4.1.1   Protective equipment - Throwaway plastic gloves,
                       apron or lab coat, safety glasses, and a lab hooo
                       adequate for radioactive work.

             4.4.1.2   Training - Workers must be trained in the proper
                       method of removing contaminated gloves and cloth-
                       ing'without contacting the exterior*'surfaces.

             4.4.1.3   Personal hygiene - Thorough washing of hands and
                       forearms after each manipulation ano before breaks
                       (coffee, lunch, and shift).

             4.4.1.4   Confinement - Isolated work area, posted with
                       signs, segregated glassware and tools, plastic-
                       backed adsorbent paper on benchtops.

             4.4.1.5   Waste - Good technique includes minimizing con-
                       taminated waste.   Plastic bag liners should be
                       used in waste cans.  Janitors must be trained in
                       the safe handling of waste.

             4.4.1.6   Disposal of wastes - 2,3,7,8-TCDD decomposes above
                       800°C.  Low level waste such as absorbent paper,
                       tissues, and plastic gloves may be burned in an
                       incinerator.  Gross quantities -{mil 1 igrams) should
                       be packaged securely and disposed through commer-
                       cial or governmental channels which are capable
                       of handling extremely toxic wastes.  Liquids
                       should be allowed to evaporate in a good hood and
                       in a disposable container.
                               A-30

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4.4.1.7   Decontamination • For personal decontamination, use
          any mild soap with plenty of scrubbing action.  For
          decontamination of glassware, tools, and surfaces,
          Chlorothene NU Solvent (trademark of the Dow Chem-
          ical Company) is the least toxic solvent shown to
          be effective.  Satisfactory cleaning may be accom-
          plished by rinsing with Chlorothene, then washing
          with any detergent and water.  Dishwater may be
          disposed to the sewer.   It is prudent to minimize
          solvent wastes because they may require special dis-
          posal through commercial sources whicn are expensive.

4.4.1.8   Laundry - Clothing known to be contaminated should
          be disposed with the precautions described unaer
          Section 4.4.1.6.  Lab coats or other clothing worn
          in 2,3,7,8-TCDD work area may be laundered.   Cloth-
          ing should be collected in plastic bags.   Persons
          who convey the bags and launder the clothing should
          be advised of the hazard and trained in proper
          handling.  The clothing may be put into a washer
          without contact if the launderer knows the problem.
          The washer should be run through a cycle before
          being used again for other clothing.

4.4.1.9   Wipe tests - A useful method of determining clsan-
          liness of work surfaces and tools is to wipe the
          surface with a piece of filter paper.   Extraction
          and analysis by gas chromatography can achieve a
          limit of sensitivity of 0.1 ug per wipe.   Less
          than 1 ug of 2,3,7,8-TCDD per sample indicates
          acceptable cleanliness; anything higher warrants
          further cleaning.   More than 10 ug on a wipe
          sample constitutes an acute hazard and requires
          prompt cleaning before further use of the equip-
          ment or work space.   A high (> 10 |jg) 2,3,7,8-TCDD
          level indicates that unacceptable work practices
          have been employed in the past.

4.4.1.10  Inhalation - Any procedure that may produce  air-
          borne contamination must be done with gooa venti-
          lation.   Gross losses to a ventilation system must
          not be allowed.   Handling of the dilute solutions
          normally used in analytical work presents no in-
          halation hazards except in the case of an accident.

4.4.1.11  Accidents - Remove contaminated" clothing immedi-
          ately, taking precautions not to contaminate skin
          or other articles.   Wash exposed skin vigorously
          and repeatedly until  medical  attention is obtained.
                  A-31

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5.0  Apparatus and Materials

     5.1  Sampling containers - Amber glass bottles or jars, 1-L or other
          aopropriate volume, fitted with screw caps lined with Teflon® are
          suitable for sample containment.   Cleaned aluminum foil may be sub-
          stituted for Teflon® if the sample is not corrosive.   If amber con-
          tainers are not available, samples should be protected from light
          using foil  or an opaque outer container.   The containers should be
          washed, rinsed with acetone or methylene chloride, and driea before
          use in order to minimize contamination.

     5.2  Glassware - Glassware requirements will  be dictated by the extrac-
          tion and cleanup procedures employed by each laboratory.   Typical
          glassware requirements are given below.   All specifications are
          suggestions only.   Catalog numbers are included for illustration
          only.

          5.2.1-   Volumetric flasks - Assorted sizes.

          5.2.2   Pi pets - Assorted sizes, Mohr delivery.

          5.2.3   Micro syringes - 1.0 uL with fused silica needle for en-
                  column gas chromatographic analysis.

          5.2.4   Chromatography columns - Sizes dictated by method employed.
                  A typical  column is the Chromaflex column, 400 mm x 19 mm ID
                  (Kontes K-420540-9011) or equivalent.

          5.2.5   Kuderna-Oanish evaporative concantrator apparatus.

                  5.2.5.1   Concentrator tube - 10 ml, graduated (Kontes
                            K-570050-1025 or equivalent).  Calibration should
                            be checked.  Ground glass stopper (size 519/22
                            joint) is used to prevent evaporative losses.

                  5.2.5.2   Evaporation flask - 500 ml (Kontes K-57001-0050
                            or equivalent).  Attached to concentrator tube
                            with springs (Kontes K-662750-0012 or equivalent).

                  5.2.5.3   Snyder column - Three-ball macro (Kontes K-503000-
                            0121).

     5.3  Balance - Analytical, capable of-accurately weighing 0.0001 g.

     5.4  Gas chromatographic system.

          5.4.1  _ Gas chromatograph - An analytical system complete with a
                 "temperature programmable gas chromatograph and all  required
                  accessories including syringes, analytical columns, and
                  gases.  The injection port should be designed for on-coiumn
                  injection when using capillary columns.  Other capillary
                  injection techniques (split, splitless, etc.) may be used
                                    A-32

-------
             provided the performance specifications stated in Section
             7.1 are met.

     5.4.2   Capillary GC column - A 10 to 60 m long x 0.25 mm ID fused
             silica column with a 0.25 urn thick OB-5 (Supelco Inc.,
             Bellefonte,  Pennsylvania) liquid phase is recommended.
             Alternate liquid phases may include Silar 10-C, SP-2330,
             SP-2340, 08-225, SE-54, SP-2100, or other liquid phases
             which meet the performance specifications stated in Section
             7.1.   The Silar 10-C, SP-2330, and the SP-234Q columns are
             recommended  to achieve isomer specific analyses (e.g.,
             2,3,7,8-TCDD from the other 21 TCDD isomers).

     5.4.3   A data system should be used for manipulation of GC/ECD data.
             At a minimum, electronic integration should be used.   The
             ideal system will allow for archiving raw chromatograohic
             aata on magnetic media.

5.5  Gas chromatograph/mass spectrometer system.

     5.5.1   Gas chromatograph - As described in Section 5.4.1.

     5.5.2'   Capillary GC column - As described in Section 5.4.2.

     5.5.3   Mass spectrometer - Should be caoable of scanning a rsnge
             of 400 amu in 1.5 s or less, collecting at least five aata
             points per chromatographic peak, utilizing a 70-eV (nominal)
             electron energy in the electron impact ionization mode, and
             producing a  mass spectrum which meets all the criteria in
             Table 6 when 50 ng of decafluorotriphenyl phosphine [DFTPP,
             bis(perfluoroohenyl)phenyl phosphine] is injected througn
             the GC inlet.  Any GC to MS interface that gives acceptable
             calibration  points at the working range and achieves all
             acceptable performance criteria (Section 10) may be used.
             Direct coupling of the fused silica chromatographic column
             to the MS is recommended.  Glass interfaces, if used, can
             be deactivated by silylation with dichlorodimethylsilane
             or an equivalent silylation reagent.

     5.5.4   A computer system that allows the continuous acquisition
             and storage  on machine-readable media of all mass spectral
             data obtained throughout the duration of the chromatographic
             program should be interfaced to the mass spectrometer.  The
             data system  should have the capability of integrating the
             abundances .of the .selected ions between specified limits and
             relating integrated abundances to concentrations using the
             calibration  procedures described in these guidelines.   The
             computer should have software that allows searching any mass
             spectral data file for ions of a specific mass and plotting
             such ion abundances versus time or scan number to yield an
             extracted ion current profile (EICP).   The software should
             also allow integration of ion abundances in any EICP between
             specified time or scan number limits.
                               A-33

-------
 Table 6.  OFTPP Key Ions and Ion Abundance Criteria

m/z                      Ion abundance criteria


197                    Less than 1% of m/z 198
198                    100% relative abundance
199                    5-9% of m/z 198

275                    10-30% of m/z 198

365                    Greater than 1% of m/z 198

441                    Present but less than m/z *43
442                    Greater than 40% of m/z 198
443                    17-23% of m/z 442
                        A-34

-------
6.0  Reagents
     6.1  Solvents - All solvents should be pesticide residue analysis grade.
          New lots should be checked for purity by concentration of an aliquot
          by at least as much as will occur during the sample preparation
          procedure.  The concentrated solvent sample should then be analyzed
          in the same manner as the samples.

     6.2  Standard chlorinated and brominated congeners - Standards of the
          HDD and HOP congeners, listed in Table 7, are available from Ultra
          Scientific (Hope, Rhode Island), KOR Isotopes (Cambridge,
          Massacnusetts), and Cambridge Isotope Laboratories (Woburn,
          Massachusetts).

     6.3  Standards containing mixed brominated/chlorinated congeners of the
          HDDs and HDFs are not commercially available at this time.  If
          analyses for these congeners are to be conducted, standards must be
          synthesized and characterized as to identity and purity prior to
          sample analysis.   One standard per homolog group may be adequate to
          conduct the analysis.

     6.4  Calibration standard stock solution - Primary dilutions of each of
          the individual HDDs and HOFs which are to be used are preparea by
          weighing approximately 1 to 10 mg of material with an accuracy of
          1% or better.  The standard is then diluted to 1.0 ml, or another
          known volumn, with an appropriate solvent.   The concentration is
          calculated in milligrams per milliliter.   The primary dilutions
          should be stored at 4°C, or lower; in screw-cap vials with Teflon®
          cap liners.  The meniscus is marked on the vial wall to-monitor
          solvent evaporation.

     6.5  Working calibration standards - Working calibration standards,
          similar in HDD or HOP composition and concentration to the samples,
          are prepared by mixing and diluting the individual standard stock
          solutions.  The mixture is diluted to a known volume with the same
          solvent that is used for the final  sample extract.  The concentra-
          tion is calculated in nanograms per milliliter as the individual
          components.  Dilutions are stored at 4°C, or lower, in narrow-mouth,
          screw-cap vials with Teflon® cap liners.   The meniscus is marked on
          the vial wall to monitor solvent evaporation.   These solutions should
          be labeled with the analytes and concentration, solvent,  initials of
          the preparer, and the date.

     6.6  Alternatively, certified stock solutions, if available from a sup-
          plier, may be used in lieu of the procedure described in  Section 6.5.

     6.7  DFTPP standard - A 50-ng/uL solution of DFTPP is"prepared in acetone
          or another appropriate solvent.

     6.8  Surrogate standard solutions - These solutions should be  preoared
          and labeled as described for the calibration standards
                                    A-35

-------
        fable 7.   Currently Available HDD and HDF Standards
Unlabeled
Stable isotope labeled
Chlorinated dioxins:
Mono:
Di:


Tri:


Tetra










Penta


Hexa:


Hepta
Octa:
Bromi
Mono:
Di:

Tri:
Tetra



Penta

Hexa:
2
2,3
2,3
2,7/2,8a
1,2,3
1,2,4
2,3,7
: 1,2,3,4
1,2,3,7/1,2,3,8!
1,2,3,7/1,2,8,9?
1,2,4,7/1,2,4,8?
1,2,6, 7/l,2,8,9a
1,2,7,8
1,2,8,9
1,3,6,8
1,3,6,8/1. 3, 7,9a.
1,3,7,8
2,3,7,8
: '1,2,3,4,7
1,2,3,7,8
1,2,4,7,8
1,2,3,4,7,8
1,2,3,6,7,8
1,2,3,7,8,9
: 1,2,3,4,6,7,8
1,2,3,4,6,7,8,9
nated dioxins:
1
2,7
2,8
2,3,7
: 1,2,3,4
1,3,6, 8/1, 3, 7, 9a
1,3,7,8
2,3,7,8
: 1,2,3,7,8
1,2,4,7,3
1,2,3,4,7,8

2,7/2,8 (u-13C12, 99%)b





1,2,3,4 (13CS, 99%)
2,3,7,8 (U-13C12, 99%)
2,3,7,8 (U-37C141 96%)c
1,2,3,4 (U-13C12, 99%)








1,2,3,7,8 (U-13C12, 99%)

1,2,3,6,7,8 (IM3C12, 99%)
1,2,3,7,8,9 (U-i3C12, 99%)

1,2,3,4,6,7,8 (U-13C12, 99%)
1,2,3,4,6,7,8 (U-13C12, 99%)





2,3,7,8 (U-i.3Cl2> 99%)



1,2,3,7,8 (U-13C12, 99%)

1,2,3,4,7,8 (U-13C12, 99%)
   1,2,3,6,7,8/1,2,3,7,8,9e
                               A-36

-------
                             Table 7 (continued)
     Unlabeled                                 Stable  isotope  labeled


Hepta:  1,2,3,4,6,7,8

Octa:   1,2,3,4,6,7,3,9

Mixed 3r/Cl dioxins:

Tri:    7-8romo-2,3-dichloro

Tetra:  2,3-Qibromo-7,8-dichloro
        2,8-Oibromo-3,7-dichloro
        2-3romo-3,7,3-dichloro

Penta:  2-3romo-l,3,7,3-teT:rachioro

Hexa:   3-3romo-l,2,4,7,8-pentachloro

Chlorinated dibenzofurans:
Mono:   2
        3
        4

Oi:     2,3
        2,6
        2,7
        2,8
        4,6

Tri:    1,2,3
        1,2,4
        1,3,6
        1,3,7
        1,6,7
        2,3,4
        2,3,6
        2,3,8
        2,4,6
        2,4,7
        2,4,8
      . 2,6,7

Tetra:  1,2,3,4                                2,3,7,8 (U-13C12, 99%)
        1,2,3,6
        1,2,3,7
        1,2,3,8
        1/2,3,9
        1,2,4,6
        1,2,4,8
        1,2,4,9
        1,2,7,3
        1,3,4,5
        1,3,4,7
                                    A-37

-------
                             Table 7 (continued)
     Unlabeled
                                       Stable  isotope  labeled
Tetra:  1,3,4,8
(cont.) 1,3,4,9
        1,3,6,7
        1,3,6,8
        1,3,6,9
        1,3,7,8
        1,3,7,9
        1,4,6,7
        1,4,7,8
        2,3,4,6
        2,3,4,7
        2,3,4,8
        2,3,4,9
        2,3,6,7
        2,3,6,8
        2,3,7,8
        2,4,6,7
        2,4,6,8
        3,4,6,7
Penta:
Hexa:
1,2,3,4,6
1,2,3,4,7
1,2,3,4,8
1,2,3,4,9
1,2,3,6,7
1,2,3,7,8
1,2,3,7,9
1,2,3,8,9
1,2,4,6,7
1,2,4,7,8
1,3,4,6,7
1,3,4,7,8
2,3,4,6,7
2,3,4,6,8
2,3,4,6,9
2,3,4,7,8
2,3,4,7,9
1,2,3,4
1,2,3,4
1,2,3,4
1,2,3,6
1,2,3,6
1,2,3,7
1,2,4,6
1,2,4,6
1,3,4,6
1,3,4,6
2,3,4,6
,7,8
,7,9
,8,9
,7,8
,8,9
,8,9
,7,8
,8,9
,7,8
,7,9
,7,8
                                1,2,3,7,8 (U-i3C12) 99%)
1,2,3,4,7,8 (U-«C12, 99%)
                                      A-38

-------
Table 7 (concluded)
Unlabeled
Hepta:
Octa:
1,
1,
1,
1,
2
2
2
2
Brominated
01:

Tri:
Tetra:
Penta:
Hexa:
2,
2,
1,
2,
1,
1,
7
3
3
3
2
2
,3,
,3,
,3,
,3,
4
4
4
4
,6
,6
,7
,s
,7,8
,8,9
,8,9
,7,3,9
Stable
1
1
,2
,2
,3,4
,3,4
isotope labeled
,6
,6
,7,8 (U-13C12, 99%)
,7,3,9 (U-13C12, 99%)
dibenzofurans:


,3
,7,
,3,
,3,



8
7
4




,8
,7





,8



2
1
1



,3
,2
,2



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






(U-13C12> 99%)
,8
,7
(U-13C12, 99%)
,8 (U-13C12> 99%)
Mixed 3r/Cl furans:

Tri:     8-8romo-2,3-dichloro

Tetra:   6,3-Oibromo-2,3-dichloro
        8-Bromo-2,3,4-trichloro
Penta:   6,8-Oibromo-2,3,4-trichloro
 Mixture of indicated isomers.   The relative amounts are unknown; therefore,
.these standards are not useful for quantitation calibration.
 U-13C12 indicates that the compound is universally labeled with carbon-13.
 U-37C14 indicates that the compound is universally labeled with chlorine-37.
                                     A-39

-------
     6.9  Internal  standard solution - The chosen internal-standard-is-pre-
          pared at a nominal  concentration of 1 to 10 mg/mL in an appropriate
          solvent.   The solution is further diluted to give a working standard.

     6.10 Solution stability - The calibration standard, surrogate, internal
          standard, and OFTPP solutions should be checked frequently for
          stability.   These solutions should be replaced after 6 months, or
          sooner if comparison with quality control check samples indicates
          compound degradation or concentration change.


7.0  Calibration

     The guidelines presented in this section are based on calibration using
     surrogates and internal  standards.  Isotope dilution techniques may also
     be applicable and may be substituted if validated prior to use.

     7.1  The gas chromatograph should meet the minimum operating parameters
          shown in Tables 8 or 9 on a daily basis.  If all criteria are not
          met, the analyst should adjust conditions until all criteria are
          met.

     7.2  The electron capture detector-should meet the manufacturer's
          specifications on a daily basis.   The manufacturer's procedure for
          determining detector performance should be usea.  If all performance
          criteria are not met, corrective action, as specifiea by the manu-
          facturer, should be taken and the procedures repeated until  the
          detector is within specifications.

     7.3  The mass spectrometer should meet the minimum operating parameters
          shown in Tables 10 or 11 on a daily' basis.  If all criteria are
          not met,  the analyst should retune the spectrometer and repeat the
          test until  all conditions are met.

     7.4  If the analytical system (HRGC/MS, HRGC/ECD) has not been demon-
          strated to yield a linear response or if the analyte concentrations
          are more than one order of magnitude different from those in the
          RF solution, a calibration curve should be prepared.  If the ana-
          lyte and RF solution concentration differ by more than two orders
          of magnitude, a calibration curve must be prepared.  A calibration
          curve should be established with triplicate determinations at three
          or more concentrations bracketing the analyte levels.
                                    A-40

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     Table 8.   Recommended Operating Parameters for High Resolution Gas
         Chromatographic System for Use with Mass Spectral  Detection
Parameter
Recommended
Tolerance
Gas chromatograph
Column

Liquid phase

Liquid phase thickness
Carrier gas
Carrier gas velocity
Injector
Separator
Transfer line temperature
Injector temperature
Initial column temperature
Column temperature program
Finnigan 9610
15 m x 0.25 mm ID
  fused silica
08-5 (J&W)

0.25 urn
Helium
30 cm/sec
On-column (J&W)C
Noned
320°C
Optimum performancec
80°C (2 min)f
80°C to 320°C at
  10°C/min
Other0
Other

Other nonpolar or
  semioolar
< 1 urn
Nitrogen or hydrogen
Optimum performance
Other
Glass jet or other
Optimum8
Optimum performance
Other
Other
Injection volume
Tailing factor3
Peak widthh
1.0 uLc
0.7 to 1.5
7 to 10 sec
Other
0.4 to 3
< 15 sec
.Substitutions permitted if performance criteria are met.
cMeasured by injection of air at 30°C column temperature.
 For on-column injection, manufacturer's instructions should be followed
dregarding injection technique.
 Fused silica columns may be routed directly into the ion  source to prevent
eseparator discrimination and losses.
 High enough to elute all analytes, but not high enough to degrade the column
fif routed through the transfer line.
 With on-column injection, the initial  temperature is dependent on the
 boiling point of the solvent.
9Tailing factor is the width of the front half of the peak at 10% height
 divided by the width of the back half  of the peak at 10%  height for single
^analytes in the calibration standard solution.
 Peak width at 10% height for single analytes in the calibration standard
 solution.
                                     A-11

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     Table 9.   Recommended Operating Parameters for High Resolution Gas
        Chromatographic/Electron Capture Detection (HRGC/ECD) System
Parameter
Recommended
Tolerance
Gas chromatograph

Detector

Column


Liquid phase


Liquid phase thickness

Carrier gas

Detector make-up gas

Carrier gas velocity

Injector

Injector temperature

Detector temperature

Initial column temperature

Column temperature program


Injection volume

Tailing factor6

Peak widthf
Varian 3700

63Ni; pulsed

15 m x 0.25 mm ID
  fused silica

08-5 (J&W)


0.25 urn

Helium

Nitrogen

30 cm/sac0

On-column (J&W)

Optimum performance

350°C

80°C (2 min)d

80°C to 320°C at
  10°C/min

1.0 uLC

0.7 to 1.5

7 to 10 sec
Other

Sc3H

Other
Other nonpolar or
  semi polar
Nitrogen or hydrogen

Argon/methane

Optimum performance

Other

Optimum performance

Other

Other

Other


Other

0.4 to 3

< 15 sec
.Substitutions permitted if performance criteria are met.
 Measured by injection of air at 30°C column temperature.
 For on-column injection, manufacturer's instructions should be followed
 .regarding injection technique.
 v^ith on-column injection, the initial temperature is dependent on the
 boiling point of the solvent.
 Tailing factor is the width of the front half of the peak at 10% height
 diviaed by the width of the back half of the peak at 10% height for single
^analytes in the calibration standard solution.
 Peak width at 10% height for single analytes in the calibration standard
 solution.

                                     A-42

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       Table 10.  Recommended Operating Parameters for Quadrupole Mass
                             Spectrometer System
Parameter
Recommended
Tolerance
Mass spectrometer

Data system

Scan range

Scan time

Resolution

Ion source temperature

Electron energy0

Trap current

Electron multiplier voltage

Preamplifier sensitivity
 Finm'gan 4043

 Incos 2400

 240-850 amu

 1 sec

 Unit

 280° C

 70 eV

 0.2 mA

 -1,600 V

 10"6 A/V
Other3

Other

Other

Otherb

Optimum performance

250-350°C

70 eV

Optimum performance

Optimum

Set for desired
  Corking range
bSubstitutions permitted if performance criteria are met.
 Greater than five data points over a GC peak is a minimum.
 Filaments should be shut off during solvent elution to improve instrument
 stability and prolong filament life, especially if no separator is used.
                                   A-43

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       Table 11.   Recommended Operating Parameters for Magnetic Sector
                          Mass Spectrometer System
Parameter
Recommended
Tolerance
Mass spectrometer
Data system
Scan range
Scan mode
Cycle time
Resolution
Ion source temperature
Electron energy
Emission current
Filament current
Electron multiplier voltage
Finnigan MAT 311A
Incos 2400
240-850 amu
Exponential (up)
1.2 sec
1,000
280°C
70 eV
1-2 mA
Optimum
-1,600 V
Other*
Other
Other
Other
Otherb
> 500
250-350°C
70 eV
Other
Optimum
Optimum
.Substitutions permitted if performance criteria are met.
 Greater than five data points over a GC peak is a minimum.
 Filaments should be shut off during solvent elution to improve instrument
 stability and prolong filament life, especially if no separator is used.
                                    A-44

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7.5  The HOD or HDF response factors (RFu) should be determined using
     Equation 7-1 for the analyte homologs.



                       *F  -inl^l                           E,  7-
                       R~K   a   v M                            CLi-
     where:

             RF.  - response factor of a given HDD or HDF congener

              A.  = area of the HDD or HDF congener peak

              M.  = mass of HOD or HDF congener injected (pg)

             M.  = mass of internal  standard injected (pg)

             A.  = area of internal  standard peak


     Using the same conditions as for RF. ,  the surrogate response
     factors (RF.) should be determined using Equation 7-2.

                             A  x M
                       RF  = VL^s.                        -   Eq. 7-2
                              is    s


     where:

              A  = area of the surrogate peak

              M  = mass of surrogate injected (pg)


     Other terms are the same as defined in Equation 7-1.

     If specific congeners are known to be  present and if standards are
     available, selected RF values may  be employed.

     For selected ion monitoring, the primary and secondary ions of the
     molecular clusters of both the analyte and surrogate are generally
     used to determine the RF values.   If alternate ions are to be used
     for quantisation, the RF should be determined using those charac-
     teristic ions.

     The RF  value should be determined  in a manner which ensures an
     accuracy and precision of ± 30% or better.   The most common pro-
     cedure  is to determine the RF value for at least three different
     concentration levels to establish  a working calibration curve.
                               A-45

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          Other options include, but are not limited to, triplicate deter-
          minations of a single concentration spaced throughout a day or a
          running mean (RF) based on one value per day for seven consecutive
          days.

     7.6  The relative retention time (RRT) windows for the homologs and
          surrogates should be determined.   The windows should be set wider
          than observed if all isomers are not available as standards.
8.0  Sample Collection. Handling, and Preservation

     8.1  Amber glass sample containers should have Teflon®-lined screw caps.
          If the sample is noncorrosive, aluminum foil liners, rinsed with
          methylene chloride, may be substituted.   The volume and configura-
          tion of the sample containers will be determined by the amount and
          physical properties of sample to be collected.   For dry powders,
          other containers, such as heavy-walled polyethylene bags, may be
          appropriate.

     8.2  Sample container preparation.

          8.2.1   All sample containers and caps should be washed in deter-
                  gent solution, rinsed with tap water, and then with dis-
                  tilled water.   The containers and caps  are allowed to drain
                  dry in a contaminant-free area.   The caps should be rinsed
                  with pesticide grade hexane and allowed to air dry.

          8.2.2   Sample bottles, if used, should be heated to 400°C for
                  15 to 20 min or rinsed with pesticide grade acetone or
                  hexane and allowed to air dry.

          8.2.3   The clean bottles should be stored inverted or sealed until
                  use.

     8.3  Sample collection.

          8.3.1   The primary consideration in sample collection is that the
                  sample collected is representative of the whole.   Therefore,
                  sampling plans or protocols for each individual producer's
                  situation should be developed.  The recommendations pre-
                  sented here describe general situations.  The number of
                  replicates and sampling frequency should also be planned
                  prior to sampling.   Guidelines for the  collection of repre-
                  sentative samples are provided as Appendices C and 0.

          8.3.2   Discrete product units - If the product is small  enough
                  that one or more discrete units will be used as the analyt-
                  ical  sample, a statistically random sampling approach is
                  recommended.
                                    A-46

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          8.3.3   Liquids or free-flowing solids - If possible, the source
                  is mixed thoroughly before collecting the sample.  If mix-
                  ing is impractical, the sample should be collected from a
                  representative area of the source.   If the liquid is flow-
                  ing through an enclosed system, sampling through a valve
                  should be randomly timed.

          8.3.4   Solids - Larger bulk solids which should be subsampled to
                  get a reasonably sized analytical sample should be treated
                  on a case-by-case basis.   A representative sample should
                  be obtained by designing a sampling location selection
                  scheme such that all parts of the whole have a finite known
                  probability of inclusion.

     3.4  Sample preservation - Product samoles should be stored as the bulk
          or packaged product inventory would be stored, or protected from
          light in a cool, dry area.   Intermediates,  process samples, or
          other nonproduct specimens should be stored protecteo from light
          at 4°C or lower.
9.0  Sample Preparation

     Since a wide variety of matrices may be subjected to analysis by thesa
     guidelines,  exact extraction/cleanup procedures cannot be specified.
     Each laboratory should prepare a method document for the specific
     analysis to  be performed.

     This section describes general guidelines for subsampling,  addition of
     surrogates,  dilution, extraction, cleanup,  extract concentration, and
     other sample preparation considerations.   Only those methods that deal
     directly with commercial products have been presented in detail.  Options
     for analysis are not limited to those presented herein.   Additional pro-
     cedures may  be referenced from several analytical protocols dealing with
     analyses of  chlorinated dibenzodioxins and furans in environmental
     matrices.

     Standard analytical  methods are available for the analysis  of 2,3,7,8-
     TCDD in soils and sediments and water/wastewater and hazardous wastes
     (EPA Method  613 1984; RCRA Method 8280, EPA/ERD-L 1984;  EPA Region  VII
     1983; Harless et al.  1980; McMillin et al.  1983).  These methods have
     been validated for the measurement of 2,3,7,8-TCDD to detection levels
     of 1 ng/g (ppb) for soils and sediments and 1 ng/L (ppt) for water/
     wastewater.   The soil and sediment protocol has been used for the anal-
     ysis of over 11,000 samples for 2,3,7,8-TCDD in EPA Region  VII alone.
     These methods have not been validated for the analysis'- of total PCDDs
     and PCDFs.   However,  several laboratories have provided  sufficient  docu-
     mentation to suggest that modification of these procedures  can extend
     the analysis for total tetrachloro- through octachloro-PCDDs and PCDFs.
     RCRA Method  8280 is  currently being validated for the analysis of tetra-
     through hexachloro-PCDDs and PCDFs in hazardous wastes.
                                    A-47

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The extraction and cleanup procedures for the determination of PCDOs and
PCDFs in phenoxyalkanoic herbicides has been reviewed by Baker et al.
(1981).   Extraction techniques have included steam distillation of TCOD
from an alkaline solution of technical 2,4,5-T (Brenner et al. 1972,
1974), hexane extraction from an alkaline solution of 2,4,5-T, liquid-
liquid partitioning of TCOD into hexane from a solution of technical
2,4,5-T in dimethylformamide/acetonitrile/water (Vogel 1976), and sepa-
ration of TCDD from isobutoxy methyl esters of 2,4,5-T and fenoprop using
a silica gel column (Ramsted et al.  1977).

Woolson et al. (1972) evaluated 17 different pesticides containing the
polychlorophenoxy group for the presence of PCDOs.  Extraction was accom-
plished using hexane extraction from methanolic potassium hydroxide solu-
tion.  Cochrane et al.  (1981) described the analysis of technical and
formulated products of 2,4-0 as the ester, free acid, and the amine.  The
2,4-0 esters were transferred directly onto a silica column and eluted
with 30% dichloromethane in hexane.   The 2,4-D acids were dissolved in
acetonitrile/water and the PCDOs were partitioned into hexane.  The 2,4-0
amines were dissolved in water and partitioned into hexane.

Analysis of pentachlorophenol (PCP) for PCODs has been reported by sev-
eral researchers (Buser 1975, Buser and Bosshardt 1976, Mieure et al.
1977, Blaser et al. 1976, Firestone et al. 1972,  Plimmer et al.  1973,
Lamberton et al. 1979,  Nelsson and Renberg 1974,  and Pfeiffer et al.
1978).  The analytical  methods range from dissolution in a polar solvent
and partitioning the PCDOs into a nonpolar solvent to separation from
the commercial product using a macro siljca adsorption column (Mieure
et al. 1977).  Yamagishi et al. (1981) reported a similar procedure for
the determination of PCDOs and PCDFs in commercial diphenyl  ether herbi-
cides such as nitrophen and chloromethoxynil.

9.1  Sample homogenization and subsampling - The sample is homogenized
     by shaking, blending, shredding, crushing, or other appropriate
     mechanical techniques.  A representative subsample of an appropri-
     ate mass is then taken.   The sample size is dependent upon the
     anticipated HDD or HDF levels and difficulty of the subsequent
     extraction/cleanup steps.

9.2  Surrogate addition - An appropriate volume of surrogate solution
     is pipetted into the sample.   The final concentration of the surro-
     gates should be in the working range of the calibration curve and
     well above the matrix background.  The surrogates are thoroughly
     incorporated by further mechanical agitation.  For nonviscous
     liquids, shaking for 30 s should be sufficient.   For viscous
     liquids or free-flowing solids, tumbling for 10 min is recommended.
     In cases where adequate incorporation may be difficult, such as
     solids, overnight equilibration with agitation is recommended.

     Note:  The volume measurement of the spiking solution is critical
     to the overall method precision.   The analyst should exercise cau-
     tion that the volume is known with an accuracy of 13» or better.
     Where necessary, calibration of the pipet is recommended.
                               A-48

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9.3  Sample preparation (extraction/cleanup) - After addition of the
     surrogates, the sample is further treated at the discretion of the
     analyst, provided that the instrumental response of the surrogates
     meets the criteria listed in Section 7.0.  Several possible tech-
     niques are presented below for guidance only.   The applicability
     of any of these techniques to a specific sample matrix should be
     determined by the precision and accuracy of the surrogate recover-
     ies, as discussed in Section 15.2.

     9.3.1   Removal of matrix - The techniques used for separation of
             HDDs and HDFs from the sample matrix will  depend on the
             physical and chemical properties of the commercial procuct.
             The properties utilized in this scneme are volatility,
             solubility, acidity, polarity, and molecular geometry.  If
             the product is sufficiently volatile,  evaporation may be
             employed.   Differential solubility in immiscible solvents
             may be the basis for a liquid-liquid extraction.   For ex-
             ample, a salt may be dissolved in aqueous  solution and the
             HDDs and HDFs extracted with an immiscible nonpolar solvent
             such as hexane.   The solvent, number of extractions,
             sol vent-to-sample ratio, and other parameters are chosen
             at the analyst's discretion.  The planarity of the HDDs
             and HDFs may also be exploited to separate them from a
             nonplanar matrix using grapnitized carbon  column chromatog-
             raphy.  Column c.hromatography with alumina, silica, or ion
             exchange resins  may also be applicable for use with some
             matrices.   Other separation techniques may also be used if
             validated prior  to sample analysis.   Several illustrative
             examples,  taken  from the literature, are presented below.

             9.3.1.1   Extraction of chlorophenols with sodium hydroxide
                       (Firestone 1977).

                       9.3.1.1.1  Weigh 25 g of sample  into a 2-L
                                  Erlenmeyer flask.  Add 1 L of water
                                  and 200 mL of 1 N sodium hydroxide.

                       9.3.1.1.2  If necessary, warm the flask on a
                                  steam bath until  the  sample dissolves

                       9.3.1.1.3  Add 350 ml of petroleum ether ana
                                  shake the flask vigorously for about
                                  1 min.

                       9.3.1.1.4  Drain the lower layer and transfer
                                  the upper layer to a' second separa-
                                  tory funnel.

                       9.3.1.1.5  Repeat the extraction with petroleum
                                  ether twice more and  comoine the
                                  extracts.
                               A-49

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          9.3.1.1.5  Wash the combined extracts with 100 mL
                     of water by gently swirling.   Repeat
                     the wash twice with 100-mL portions
                     of water.   (Wash water should be
                     neutral.)

          9.3.1.1.7  Transfer the extract to a 2-L Erlen-
                     meyer flask and dry by adding 200 g
                     of anhydous sodium sulfate.   Swirl
                     the flask  vigorously and allow it
                     to stand for 30 min.

          9.3.1.1.8  Transfer the dry petroleum ether to
                     a second Erlenmeyer flask.   Rinse the
                     first Erlenmeyer flask twice with
                     200-mL portions of petroleum ether
                     and transfer the rinsings to the
                     second Erlenmeyer flask.

          9.3.1.1.9  Evaporate  the petroleum ether to
                     about 500  mL.

          9.3.1.1.10 Transfer to a 500-mL Kuderna-Oam'sh
                     concentrator and evaporate to about
                     10 mL.

9.3.1.2   Extraction of pentachlorophenol  with lithium
          hydroxide (Buser 1975, 1976).

          9.3.1.2.1  Add 4.0  g  of sample to 30 mL of meth-
                     anol in  a  250-mL separatory funnel.

          9.3.1.2.2  Add 10 mL.of 2.5 N lithium hydroxide
                     and 100  mL of water.

          9.3.1.2.3  Extract  with 40 mL of petroleum ether.

          9.3.1.2.4  Drain the  lower phase (aqueous) after
                     checking for alkalinity (pH > 12).

          9.3.1.2.5  Wash the organic phase with 50 mL of
                     water.   Check the last washing for
                     neutrality.

          9.3.1.2.6  Dry the  organic phase over anhydrous
                     sodium sulfate.

          9.3.1.2.7  Concentrate a 20-mL aliquot of the
                     dry organic phase to about 2 mL using
                     a stream of nitrogen.
                  A-50

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9.3.1.3   Extraction of 2,4-0 amines (Cochrane 1981).

          9.3.1.3.1  Transfer 2.0 to 2.5 g of sample to a
                     250-mL flask.

          9.3.1.3.2  Add 100 ml of water and a clean mag-
                     netic stirring bar.  Mix the solution
                     thoroughly for 10 min on a magnetic
                     stirrer.

          9.3.1.3.3  Transfer the solution to a 250-mL
                     separatory funnel and extract three
                     times with 25-mL portions of hexane.
                     Combine the extracts.

          9.3.1.3.4  Wash the hexane extracts twice with
                     20-mL portions of water.

          9.3.1.3.5  Dry the hexane by passage through
                     15 g of prewashed sodium sulfate.

          9.3.1.3.6  Concentrate to 1 to 2 ml.

9.3.1.4   Extraction of 2,4-0 acids (Cochrane 1381).  .

          9.3.1.4.1  Dissolve a 10-g sample in 400 ml of
                     acetonitnle-water (1:1) in a 500-mL
                     flask containing 40 ml of methanol.

          9.3.1.4.2  Transfer the solution to a 1-L sepa-
                     ratory funnel  using two rinses of
                     50 mL of hexane to wash the flask.

          9.3.1.4.3  Extract three times with 100-mL por-
                     tions of hexane.   Combine the extracts.

          9.3.1.4.4  Wash the combined hexane extracts
                     twice with 50-mL portions of methanol-
                     water (1:1).   Discard the rinsings.

          9.3.1.4.5  Dry the hexane extract by passage
                     through 30 g of prewashed sodium
                     sulfate in a funnel.

          9.3.1.4.6  Reduce the volume to 1 to 2 ml under
                     reduced pressure.

9.3.1.5   Extraction of 2,4-0 esters (Cochrane 1981).

          9.3.1.5.1  Activate silica gel (M.  Kieselgel  60;
                     Merck, Darmstadt,  G.F.R.) overnight
                     at 125°C.
                  A-51

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          9.3.1.5.2  Add 50 g of silica gel to a glass
                     chromatographic column (60 x 2.0 cm ID),

          9.3.1.5.3  Transfer 2 g of sample to the head of
                     the column.

          9.3.1.5.4  Elute the column with 150 ml of 30%
                     methylene chloride in hexane.

          9.3.1.5.5  Discard the first 30 ml of eluent.

          9.3.1.5.6  Collect the remaining eluent in a
                     250-mL round bottomed flask.

          9.3.1.5.7  Concentrate to 1 to 2 mi. on a rotary
                     evaporator for subsequent cleanup.

9.3.1.6   Removal of an acidic matrix (pentachloropnenol)
          with alumina (Mieure 1977).

          9.3.1.6.1  Heat 190 g of alumina (Fisher Adsorp-
                     tion A-540, 80-200 mesh) at 400°C
                     for 4 h.   Cool and pour i,nto a pint
                     bottle.  Pipette 10 mL of distil lea
                     water into the bottle, cap the bottle,
                     and shake for at least 10 min.   The
                     bottle cap should have an aluminum
                     liner!  The deactivated alumina has
                     a shelf life of several weeks.

          9.3.1.6.2  Place a small  amount of glass wool
                     in the bottom of a chromatographic
                     column (30 cm x 1.8 cm ID Pyrex with
                     a Teflon® stopcock).   Add benzene to
                     the column until it is half full.
                     Add 40 ml of deactivated alumina.
                     Allow the alumina to settle while
                     tapping the walls of the column.   Add
                     1 in.  of sodium sulfate to the top
                     of the alumina.  Open the stopcock
                     and adjust the benzene level  to the
                     top of the packing.

          9.3.1.6.3  Dissolve a 5-g portion of sample in
                     benzene in a 50-mL volumetric flask.
                     Sonicate if necessary to effect solu-
                     tion.   Dilute the flask to volume
                     with benzene.

          9.3.1.6.4  Pipete a 10-mL aliquot of the sample
                     solution dropwise onto the head of
                     the column.
                  A-52

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                  9.3.1.6.5  El'ite the column with benzene, col-
                             lecting the first 150 mL of eluent.

                  9.3.1.6.6  Concentrate the collected eluent to
                             an appropriate volume for subsequent
                             cleanup using a Kuderna-Qanish concen-
                             trator.

9.3.2   Cleanup - Several  cleanup techniques are described below
        as examples of procedures which may be applicable to the
        analysis of HDDs and HDFs in commercial  products.  Oeoending
        on the complexity of the sample, one or more of the tech-
        niques may be required to fractionate the HDDs and HDFs
        from interferences.  For most cleanups a concentrated
        (1-5 mL) extract should be used.

        9.3.2.1   Sulfuric acid cleanup (Firestone 1972).

                  9.3.2.1.1  Add concentrated extract (in petroleum
                             ether) to a 25-mL graduated cylinder.
                             Stopper and shake the cylinder for
                             about 30 s.

                  9.3.2.1.2  Let the layers separate and transfar
                             the petroleum ether layer to a 30-mL
                             beaker.

                  9.3.2.1.3  Rinse the cylinder with 5 mL of
                             petroleum ether and add the rinsings
                             to the beaker.

                  9.3.2.1.4  Add 1 g of sodium bicarbonate to the
                             petroleum ether solution.  Allow to
                             stand for 5 min.

                  9.3.2.1.5  Transfer the petroleum ether solution
                             to a small vial and evaporate the
                             contents to dryness at room tempera-
                             ture using a gentle stream of nitrogen.

                  9.3.2.1.6  Dissolve the residue in an appropriate
                             volume of a suitable solvent for fur-
                             ther cleanup or for final analysis.

        9.3.2.2   Alumina cleanup (Firestone 1972).

                  9.3.2.2.1  Activate 100-g portions of alumina
                             (Fisher No. A-540)  by heating 4 h at
                             260°C.  Transfer alumina, without
                             cooling, to a dry container and close
                             the container tightly.   Do not store
                             activated alumina for more than 3
                             days.
                          A-53

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          9.3.2.2.2  Quantitatively transfer the concen-
                     trated extract (in petroleum ether)
                     to the alumina column.

          9.3.2.2.3  Elute the column with 400 mL of
                     petroleum ether (Fraction 1).

          9.3.2.2.4  Elute the column with 200 ml of S%
                     diethyl ether in petroleum ether
                     (Fraction 2).

          9.3.2.2.5  Elute the column with 400 mL of 25%
                     diethyl ether in petroleum ether
                     (Fraction 3).

          9.3.2.2.6  Elute the column with 400 mL of
                     diethyl ether (Fraction 4).

          9.3.2.2.7  Transfer Fractions 3 and 4 to  Kuderna-
                     Oanish concentrators (keep separate)
                     and evaporate to about 10 ml.

          9.3.2.2.8  Remove from steam bath and evaoorate
                     just to dryness at room temperature
                     under nitrogen.

          9.3.2.2.9  Dissolve the residue in an appropriate
                     volume of a suitable solvent for fur-
                     ther cleanup or for final  analysis.

9.3.2.3   Alumina cleanup (Buser 1975,  1976).

          9.3.2.3.1  Prepare the column by adding 1.0 g of
                     dry alumina to a disposable  Pasteur
                     pi pete (15 cm x 5 mm ID) containing
                     a plug of glass wool.

          9.3.2.3.2  Quantitatively transfer the  concen-
                     trated extract onto the head of tne
                     column.

          9.3.2.3.3  Elute the column with 10 mL  of 2%
                     methylene chloride in hexane.   Dis-
                     card the eluent.

          9.3.2.3.4  Elute the column wi$h 10 mL  of 50%
                     methylene chloride in hexane.

          9.3.2.3.5  Adjust the sample volume and sample
                     composition as needed for further
                     cleanup or for final  analysis.
                  A-54

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9.3.2.4   Basic aluminum oxide cleanup (Mieure 1977).

          9.3.2.4.1  Add 1.0 g of aluminum oxide (ICN
                     Aluminum Oxide W 200 basic, activity
                     grade Super 1, used as received) to
                     a disposable Pasteur pipet (15 cm x
                     0.5 cm 10) containing a glass wool
                     plug.   Settle the packing by gently
                     tapping 15 to 20 times with a small
                     spatula.

          9.3.2.4.2  Quantitatively transfer 0.30 mL of
                     benzene concentrate to the column.

          9.3.2.4.3  Elute the column with 10 mL of 2%
                     methylene chloride in hexane.   Dis-
                     card the eluent.

          9.3.2.4.4  Elute the column with 10 ml of 50%
                     methylene chloride in hexane.

          9.3.2.4.5  Adjust the sample volume and solvent
                     composition as needed for instrumental
                     analysis.

9.3.2.5   Basic alumina cleanup (Cochrane 1981).

          9.3.2.5.1  Place a small glass wool plug in the
                     bottom of a glass chromatographic
                     column (25 x 1.5 cm ID).  Add 0.5 cm
                     of Ottawa standard sand.

          9.3.2.5.2  Add 15 g of basic alumina (Activity I;
                     Woelm, Eschwege,  G.F.R., used as re-
                     ceived) to the column, then top with
                     0.5 cm of sodium sulfate.

          9.3.2.5.3  Quantitatively transfer 1 to 2 mL of
                     the concentrated sample extract onto
                     the head of the column.

          9.3.2.5.4  Elute the column with (1)  100 mL of
                     hexane; (2) 100 mL of 2% methylene
                     chloride in hexane; (3) 100 mL of 5%
                     methylene chloride in hexane;  and
                     (4) 100 mL of 10% methylene chloride
                     in hexane.

          9.3.2.5.5  Discard Fractions (1) and  (2).

          9.3.2.5.6  Concentrate Fractions (3)  and (*) to
                     1 to 2 mL,  transfer to a 5-mL gradu-
                     ated centrifuge tube with  three 1-mL
                  A-55

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                     portions of benzene, then evaporate
                     to dryness.

          9.3.2.5.7  Dissolve the residue In an appropriate
                     volume of a suitable solvent for final
                     analysis.

9.3.2.6   Graphltized carbon cleanup (O'Keefe 1978).

          9.3.2.6.1  Transfer the extract (15 mL In meth-
                     ylene chloride) to a graphitized car-
                     bon column [40 mm x 5 mm ID, packed
                     with Carbopack AHT (Supelco, Inc. ,
                     Bellefonte, Pennsylvania)].

          9.3.2.6.2  Elute the nonplanar aromatic compounds
                     with 50 ml of benzene/aiethyl  ether
                     (40/60).   Discard the eluent.

          9.3.2.6.3  Elute the planar aromatic compounds
                     with 70 ml of pyridine.

          9.3.2.6.4  Dilute the pyridine fraction with
                     140 ml of L" HC1 and extract three
                     times with 30-mL portions of hexane.

          9.3.2.6.5  Wash the hexane extract with 100 ml
                     of water, then dry over sod-ium sulfate.

          9.3.2.6.6  Adjust the sample volume and solvent
                     composition as needed for instrumental
                     analysis.

9.3.2.7   Graphitized carbon cleanup for TCDD (EPA  1983)

          9.3.2.7.1  Prepare 18% Carbopack C on Celite
                     545® by thoroughly mixing 3.6  g of
                     Carbopack C (80/100 mesh) and  16.4
                     g of Celite 545® in a 40-mL  vial.
                     Activate at 130°C for 6 h.   Store
                     in a desiccator.

          9.3.2.7.2  Prepare a column using a standard
                     size (5-3/4 in.  long by 7.0  mm o.d.)
                     disposable pipet fitted with a small
                     plug of glass  wool...-Using a vacuum
                     aspirator attached to the pointed
                     end of the pipet,  add the Carbopack/
                     Celite mix until  a 2-cm column is
                     obtained.
                  A-56

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          9.3.2.7.3  Preelute the column with 2 ml of
                     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 of hexane.

          9.3.2.7.4  While the column is still  wet with
                     hexane, add 50 ul_ of sample extract.

          9.3.2.7.5  Elute the column sequentially with
                     two 1-mL aliquots of hexane,  1 mL
                     of 1:1 cyclohexane in methylene
                     chloride, and 1 ml of 75:20:5 meth-
                     yl ene chloride/methanol/benzene.

          9.3.2.7.6  Collect tne TCDD fraction  oy  elution
                     with 2 ml of toluene.

          9.3.* 7.7  Reduce the volume to near  dryness
                     and add isooctane to obtain a final
                     volume of 50 uL.

9.3.2.8   Silica gel  cleanup (Pfeiffer 1978).

          9.3.2.8.1  This proceaure is designed to remove
                     chlorophenols, chlorophenoxy  pnenols,
                     or other polar impurities  extracted
                     from an aqueous solution.

          9.3.2.8.2  Dry the silica gel  (Bio-Si 1 A,  100/120
                     mesh; Bio-Rad Laboratories, Richmond,
                     California) at 180°C in a  tube furnace
                     for 1.5 h under a 100 cc/min  nitrogen
                     flow.  Store the dried silica gel,
                     capped in glass bottles,  in a glass
                     desiccator containing phosphorous
                     pentoxide.

          9.3.2.8.3  Place a small plug  of glass wool  in
                     a disposable capillary pipet  (150 x
                     5 mm ID;  VWR Scientific, San  Francisco,
                     California).

          9.3.2.8.4  Add the silica gel  to the  pipet to
                     produce an adsorbent bed 60 mm in
                     height.

          9.3.2.8.5  Transfer 7.5 mL of  the hexane extract
                     onto the head of the column.
                  A-57

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          9.3.2.8.6  Elate the column with 7 mL of hexane,
                     maintaining the flow through the
                     column at 2 to 3 drops per second by
                     applying gentle air pressure at the
                     top of the column using a rubber bulb.

          9.3.2.8.7  Collect all of the eluent.  Concen-
                     trate for final analysis or further
                     cleanup as necessary.

9.3.2.9   HPLC cleanup (Pfeiffer 1978).

          9.3.2.9.1  Inject an appropriate aliquot of
                     extract onto a high pressure liauid
                     chromatographic system.  An example
                     of an appropriate system is given in
                     Table 12'.

          9.3.2.9.2  Collect the column eluent within
                     established retention windows for
                     the HDD and HDF homologs.  Figure 1
                     shows a chromatogram obtained using
                     the system outlined in Table 12.

          9.3.2.9.3  Adjust the sample volume and solvent
                     composition as necessary for instru-
                     mental analysis.

9.3.2.10  HPLC cleanup (RCRA Method 8280)

          9.3.2.10.1 Operating parameters:  Column tem-
                     perature, 30°C; column pressure,
                     172 atm.; flow rate, 1.0 mL/min; UV
                     detector, 235 nm; detector sensitiv-
                     ity, 0.02 UFS; isocratic operating
                     mode, injection via 100 |A sample
                     loop.

          9.3.2.10.2 Elution profile for 2,3,7,8-TCDD:
                     An authentic standard of 2,3,7,8-
                     TCDD demonstrated a retention time
                     of 16.0 min.  Under the above con-
                     ditions  the retention time of this
                     standard  should be verified before
                     proceeding.  Typically, collect the
                     entire column effluent, eluting be-
                     tween 16.0 and 22.0 min following
                     introduction of the sample extract
                     onto the  HPLC columns, 'into a 12.0-mL
                     concentrator tube.
                  A-58

-------
  Table 12.  Example HPLC Parameters for Sample Cleanup
                     (Pfeiffer 1978)
Pump

Injector

Detector

Analytical wavelength

Column

Column dimensions

Mobile phase

Flow rate

Column temperature

Detector sensitivity

Injection volume
Waters M-6000A

Rheodyne Model 7120

Perkin-Elmer LC-55 or LC-55T

245 nm

ODS/Zorbax (6 urn) (DuPont)

4.6 mm x 250 mm

Methanol (100%)

2.0 mL/min

Ambient

0.02 AUFS

6 uL
                          A-59

-------
                                     CU-CDD
                                               Clg-CDD
                                                      CU-CDBF
                                  Minutes
                                                       10
12
Fiaure  1.  Chromatoaram  for HPLC cleanun  by  nentachloronhenol  (?feiffer 1973)
                                  A-60

-------
9.3.2.10.3  Solvent exchange, instrument analysis,
            and sample concentration:   After the
            methylene chloride/hexane effluent is
            taken to dryness, add 0.5 ml of meth-
            ane!  to the concentrator tube.

9.3.2.10.4  Using a gentle stream of prepurified
            nitrogen and a 55°C sand bath evaporate
            the effluent to 40 uL.

9.3.2.10.5  Using a 100-uL syringe draw the residue
            solution carefully into the syringe
            until an air bubble appears in the
            barrel.  Place a second 60-uL aliquot
            of methanol into the concentrator tube,
            rotate quickly for rinsing and draw
            this  into the syringe.

9.3.2.10.6  Inject the total volume into the HPLC.

9.3.2.10.7  At the appropriate retention time,
            position a sample collection bottla to
            collect the required fraction.   AGO 5
            mL of hexane to the sample bottle.

9.3.2.10.8  Add 2 mL of 5% (w/v) sodium carbonate
            to the sample fraction collected and
            shake for 1 min.

9.3.2.10.9  Quantitatively remove the hexane layer
            (top  layer) and transfer to a micro-
            reaction vessel.  Add an additional
            5 ml  of hexane, shake 1 min, and
            transfer the solution to a concentrator
            tube.

9.3.2.10.10 A Teflon boiling chip is added and
            the concentrator tube is fitted with a
            2-ball Snyder column.

9.3.2.10.11 The sample is concentrated to near
            dryness in a 90°C water bath.

9.3.2.10.12 After cooling for 10 min,  the sample
            is evaporated just.-to dryness using a
            gentle stream of prepurified nitrogen
            and a 55°C sand bath.
        A-61

-------
                            9.3.2.10.13 Transfer the residue to a 1.0-mL
                                        microflex vial  with a small  amount
                                        of toluene.   Be very careful to
                                        transfer the entire rinse solution.

                            9.3.2.10.14 Again concentrate the residue to
                                        100 uL using a  gentle stream of pre-
                                        purified nitrogen and a 55°C sand
                                        bath.

                            9.3.2.10.15 Store the microflex vial under
                                        refrigeration until just prior to
                                        GC/MS analyses.

     9.4  Miscellaneous cleanuo procedures.

          Other cleanup procedures may be employed if they are validated
          prior to use.  Possibilities include,  but are not limited to,
          extraction cartridges (e.g., Sep-Pak,  Waters  Associates),  distil-
          lation,  and thin--layer chromatography.


10.0  Gas Chromatoqraphic/Electron Impact Mass Spectrometric Determination

      10.1  Internal standara addition - An appropriate volume of the internal
            standard solution is pipeted into the sample.  The final concen-
            tration of the internal standard should be  in the working range
            of the calibration and well above the matrix background.  The in-
            ternal standard is thoroughly incorporated  by mechanical agitation.

            Note:   The volumetric measurement of the internal standard solu-
            tion is critical to the overall method precision.  The analyst
            should exercise caution that the volume is  known with an accuracy
            of 1% or better.  Where necessary, calibration of the pi pet is
            recommended.

      10.2  Tables 8, 10, and 11 summarize the recommended operation conditions
            for analysis'.  Figures 2 and 3 present, as  examples, chromatograms
            (HRGC/SIM, HRGC/FMS) obtained for an analysis of technical grade
            pentachlorophenol.

      10.3  While the highest available chromatographic resolution  is not a
            necessary objective of this protocol, good chromatographic per-
            formance is recommended.  With an optimized HRGC system, the
            probability that the chromatographic peaks  consist of single
            compounds  increases, making qualitative and quantitative data
            reduction more reliable.
                                    A-62

-------
a\
 86.6-1


  304-



100.0-


  306-



 49.5-


  308-



332.5-


  RIC-
                    T	1	T
                               A    .A
                                                 III
                                                                                 I  I   I   I
                                                                                                798720.

                                                                                                304.091
                                                                                                ± 0.500
                                                                                                922624.

                                                                                                306.092
                                                                                                ± 0.500
                                                    456704.

                                                    308.092
                                                    ± 0.500
                                                                                                3067900.
             450
             12:35
                    500
                    13:59
550
15:23
                                                                    I   I   I
                                                                                 I   I   I   I
600
16:46
650
18:10
700    SCAN
19:34  TIME
               Fiqure 2.  HRCC/SIH determination  of  tetrachlorodiberizofuran in neiiLachlorooheiiol
                                         (Midwest Research Institute).

-------
100.
  RIO

5
4
M
1
j
JjC '-J
i -



u
6






200 400 600 800 1000 1200 1400 16
5:00 10:00 15:00 20:00 25:00 30:00 35:00 40
                                                                               SCAN
                                                                               TIME
       Peak No.            Impurity
                Hydrocarbon Impurity
          2     Hexachlorodibenzofuran
          3     Hexachlorodiberrzodioxin
          4     Heptachlorodibenzofuran
          5     Heotachlorodibenzodioxin
               f Octachlorodibenzofuran
               1 Octachlorodibenzodioxin


        Figure 3.   HRGC/FMS analysis  of PCDDs and PCDFs  (C16-C18) in
               pentachlorophenol (Midwest Research  Institute).
                                     A-64

-------
      10.4  After performance of the system has been certified for the day
            and all  instrument conditions set according to Tables 3, 10, and
            11, inject an aliquot of the sample onto the GC column.   If the
            response for any ion, including surrogates and internal  standards,
            exceeds  the working range of the system, dilute the sample and
            reanalyze.   If the responses of surrogates, analytes, or internal
            standard are below the working range,  recheck the system perfor-
            mance.   If necessary, concentrate the  sample and reanalyze.

      10.5  Record all  data on a digital storage device (magnetic disk, tape,
            etc.) for qualitative and quantitative data reduction as discussed
            in Sections 12 and 13.
11.0  Gas Chromatographic/Electron Capture Detection Screening

      11.1  Internal  standard addition - An appropriate volume of the internal
            standard  solution is pipeted into the sample.   The final  concen-
            tration of the internal  standard should be in  the working range
            of the calibration and well  above the matrix background.   The in-
            ternal standard is thoroughly incorporated by  mechanical  agitation.

          " Note:   The volumetric measurement of the internal standard solution
            is critical  to the overall method precision.   The analyst should
            exercise  caution that the volume is known with an accuracy of 1£
            or better.   Where necessary, calibration of the pipet is  recommended.

      11.2  Table  9 summarizes the recommended operating conditions for analy-
            sis.   Figure 4 presents,  as  an example, a chromatogram obtained
            for an analysis of PCDOs  and PCDFs in a sample of technical  grade
            pentachlorophenol.

      11.3  While  the highest available  chromatographic resolution is not a
            necessary objective of this  protocol, good chromatographic perfor-
            mance  is  recommended.   With  an optimized HRGC  system, the proba-
            bility that the chromatographic peaks consist  of single compounds
            increases,  making qualitative and quantitative data reduction more
            reliable.

      11.4  After  performance of the  system has been certified for the day
            and all instrument conditions are optimized,  inject an aliquot of
            the clean sample extract  onto the GC column.   If the response for
            any peak,  including surrogates and internal  standards, exceeds the
            working range of the system, dilute the sample and reanalyze.   If
            the responses of surrogates, analytes,  or internal  standard  are
            below  the working range,  recheck the system performance.   If nec-
            essary, concentrate and reanalyze the sample.

      11.5  If possible, record all  data on a digital storage device  (magnetic
            disk,  tape,  etc.) for qualitative and quantitative data reduction
            as discussed in Sections  12  and 13.
                                    A-65

-------
   . JiW I.«J ,U(IU 4^.
   4i-i_.., O|>
III- li.kt,..., • j>/4-
.<4u» li|MMta
, .....*,, i r-t.j-,,,
  'MM t,j.4 u J, I U ^( U 4. J
     ... lit . |0 (A.,,. »„!( St.). I
                 Figure  4.   HRGC/ECD profile of PCDDs and  PCDFs  In pentachlorophenol
                                        (Midwest  Research Institute).

-------
12.0  Qualitative Identification

      12.1  Selected ion monitoring (SIM) or limited mass scan (LMS) data -
            The identification of a compound as a given HOD or HDF homolog
            requires that two criteria be met:

            12.1.1  (1) The peak should elute within the retention time window
                    set for that homolog; and (2) the ratio of two ions ob-
                    tained by SIM (Tables 13 and 14) or by LMS (Tables 15 and
                    16) should match the theoretical ratio within 20%.  The
                    analyst should search the higher mass windows to prevent
                    misidentification of a HOD or HOF fragment ion cluster
                    as the molecular ion.

            12.1.2  If either of these criteria is not met, interferences may
                    have affectea tne results and a reanalysis using full
                    scan EIMS conditions is recommended.

      12.2  Full mass scan (FMS) data.

            12.2.1  The peak should elute within the retention time window
                    set for that homolog.

            12.2.2  The unknown spectrum should be compared with an authentic
                    HDD or HDF.   The intensity of the three largest ions in
                    the molecular ion cluster should match tne theoretical
                    ratio within 20%.  Fragment clusters with proper abundance
                    ratios should also be present.

            12.2.3  Alternatively, a spectral search may be used to automatic-
                    ally reduce the data.  The criteria for acceptable identi-
                    fication should include a high index of similarity.

      12.3  Electron capture detection data.

            12.3.1  The peak should elute within the retention time window
                    set for that homolog.

            12.3.2  Any peak which elutes within the retention time window
                    set for a given homolog should be identified and quanti-
                    tated as a HDD or HOF.

      12.4  Disputes in interpretation - Where there is reasonable doubt as
            to the identity of a peak as a HDD or HDF, the analyst should
            either identify the peak as a HDD or HDF or proceed to a con-
            firmation analysis.
                                    A-67

-------
Table 13.   Characteristic Selected Ion Monitoring (SIM) Ions for HDDs
Homokig
C12H7C102
C12HSC1202
C12H5C1302
C12H4C1402
C12H3C1502
C12H2C1602
C12HC1702
C12C1802
C12H78r02
C12H6Br202
'-12^5^r3^2
C12H4Br402
C12H3Brs02
C12H2Br602
Cl2HBr702
C128r302
C12H6CTBr02
Cl2H5Cl2Br02
C12H4Cl3Br02
C12H3Cl48r02
C12H2ClsBr02
C12HCl6Br02
C12Cl7Br02
C12H5ClBr202
C12H4Cl2Br202
C12H3C13Br202
C12H2Cl4Br20
C12HClsBr202
C12ClsBr202
Cl2H4ClBr302
C12H3Cl2Br302
C12H2Cl3Br302
C12HCl48r302
C12Cl3Br302
C12H3ClBr402
C12H2Cl28r402
C12HCl3Br402
C12Cl4Br402

Primary
218 (100)
252 (100)
286 (100)
322 (100)
356 (100)
390 (100)
424 (100)
460 (100)
262 (100)
342 (100)
420 (100)
500 (100)
578 (100)
658 (100)
736 (100)
816 (100)
298 (100)
332 (100)
366 (100)
400 (100)
436 (100)
470 (100)
504 (100)
376 (100)
410 (100)
446 (100)
480 (100)
514 (100)
548 (100)
456 (100)
490 (100)
524 (100)
558 (100)
594 (100)
534 (100)
568 (100)
604 (100)
638 (100)
Ion (relative abundance)
Secondary
220 (34)
254 (56)
288 (99)
320 (76)
358 (56)
392 (32)
426 (98)
458 (87)
264 (99)
340 (51)
422 (98)
498 (68)
580 (98)
660 (76)
738 (98)
814 (82)
296 (76)
330 (61)
368 (66)
402 (85)
434 (97)
468 (83)
506 (77)
378 (71)
412 (90)
444 (91)
478 (78)
516 (77)
550 (90)
454 (85)
488 (73)
526 (78)
560 (92)
592 (96)
536 (81) •-•
570 (95)
602 (93)
636 (83)

Tertiary
.
256 (11)
290 (33)
324 (55)
354 (51)
394 (36)
428 (54)
462 (55)
-
344 (50)
418 (34)
502 (66)
582 (49)
660 (74)
740 (58)
813 (79)
300 (25)
334 (d6)
364 (51)
398 (44)
438 (53)
472 (65)
502 (73)
374 (4*)
414 (38)
448 (51)
482 (64)
512 (68)
552 (49)
458 (50)
492 (64)
528 (33)
562 (58)
596 (63)
532 (60)
572 (48)
606 (62)
640 (73)
                                   A-68

-------
                             Table 13 (concluded)
  Homolog
                                         Ion (relative abundance)
 Primary
Secondary
Ternary
C12H2ClBr502
C12HCl2BrsQ2
Cl2HClBr602
C12Cl28r602

Cl2ClBr702
614 (100)
648 (100)
682 (100)

692 (100)
726 (100)

772 (100)
612 (89)
546 (80)
684 (84)

594 (85)
723 (96)

770 (92)
616 (62)
650 (73)
680 (72)

590 (69)
730 (58)

774 (70)
                                    A-69

-------
Table 14.   Characteristic Selected Ion Monitoring (SIM) Ions  for HDFs
Homo log
C12H7C10
C12H6C120
C12HSC130
C12H4C140
C12H3C150
C12H,C1S0
C,,HC170
C^ClgO
C12H73rO
Cl2Hs8r20
C12HsBr30
C12H4Br40
C12H3BrsO
Cl2H28rsO
C12HBr70
C128r80
C12HsClBrO
C12H5Cl28rO
C12H4Cl3BrO
Cl2H3Cl4BrO
C12H2Cl5BrO
C12HC168rO
C12Cl7BrO
C12H5ClBr20
Cl2H4Cl2Br20
C12H3Cl3Br20
C12H2Cl48r20
C12HCls8r20
C12ClsBr20
Cl2H4ClBr30
C12H3Cl2Br30
C12H2Cl3Br30
C12HCl48r30
C12Cl58r30
C12H3ClBr40
C12H2Cl2Br40
C12HCl38r40
C12Cl4Br40

Primary
202 (100)
236 (100)
270 (100)
306 (100)
340 (100)
374 (100)
408 (100)
444 (100)
246 (100)
325 (100)
404 (100)
484 C100)
562 (100)
642 (100)
720 (100)
800 (100)
282 (100)
316 (100)
350 (100)
384 (100)
420 (100)
454 (100)
488 (100)
360 (100)
394 (100)
430 (100)
464 (100)
498 (100)
532 (100)
440 (100)
474 (100)
508 (100)
542 (100)
578 (100)
518 (100)
552 (100)
588 (100)
622 (100)
Ion (relative abundance)
Secondary
204 (34)
238 (66)
272 (99)
304 (76)
342 (66)
376 (82)
410 (98)
442 (87)
248 (99)
324 (51)
406 (98)
482 (68)
564 (98)
640 (76)
722 (98)
798 (82)
280 (76)
314 (61)
352 (66)
386 (85)
418 (98)
452 (83)
490 (76)
362 (70)
396 (90)
428 (92)
462 (78)
500 (77)
534 (90)
438 (85)
472 (73)
510 (78)
544 (92)
576 (96)
520 (80) -:
554 (94)
586 (92)
620 (83)

Tertiary
_ —
240 (11)
274 (33)
308 (56)
338 (61)
372 (51)
412 (54)
446 (56)
_
328 (50)
402 (34)
486 (66)
560 (51)
644 (74)
718 (61)
802 (78)
284 (25)
318 (*6)
348 (51)
382 (44)
422 (53)
456 (65)
486 (73)
358 (44)
392 (38)
432 (50)
466 (6^)
496 (69)
530 (61)
442 (49)
476 (64)
506 (64)
540 (58)
580 (63)
516 (60)
550 (54)
590 (62)
624 (73)
                                  A-70

-------
Table 14 (concluded)
  Homolog
            Ion (relative abundance)
 Primary
Secondary
                                      Tertiary
C12H2ClBr50
C^HCUBrsO
C12Cl38r30

C12HC18rsO
C12Cl28rsO

Cl2ClBr70
598 (100)
632 (100)
666 (100)

576 (100)
710 (100)

756 (100)
596 (89)
630 (80)
668 (84)

678 (85)
712 (96)

754 (92)
                                      600 (62)
                                      634 (73)
                                      664 (72)

                                      574 (69)
                                      708 (54)

                                      758 (69)
       A-71

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Table 15.   Limited Mass Scanning (IMS) Ranges for HDDs


 Homolog                                    Mass range


 C12H7C102                                  218 - 220
 C12H6C1202                                 252 - 256
 C12HSC1302                                 286 - 290
 C12H4C1402                                 320 - 326
 C12H3C1502                                 354 - 360
 Cl2HoCl608                                 388 - 394
 C12HC1702                                  422 - 430
 C12C1302                                    456 - 464

 C12H78r02                                  262 - 266
 C12H68r202                                 340 - 344
 C12HsBr30,                                 418 - 424
 C12H43r402                                 495 - 504
 C12H3Br302                                 574 - 584
 C12H2Brs02                                 654 - 662
 C12HBr702                                  732 - 742
 C128r802                                   • 810 - 822

 C12H6ClBr02                                 296 - 300
 C12H5Cl23r02                               330 - 334
 C12H4Cl3Br02                               364 - '370
 C12H3Cl4Br02                               398 - 404
 C12H2Cl5Br02                               432 - 440
 C12HClsBr02                                 466 - 474
 C12Cl78r02                                 500 - 510

 C12H5C18r202                               37* - 380
 C12H4Cl2Br202                              408 - 414
 C12H3Cl3Br202                              442 - 450
 C12H2Cl4Br202                              476 - 484
 C12HClsBr202                               510 - 518
 C12Cl6Br202                                 544 - 554

 C12H4C18r302                               452 - 458
 C12H3Cl28r30,                              486 - 494
 C12H2Cl38r302                              520 - 528
 C12HCl48r302                               554 - 562
 C12Cl5Br302                                 588 - 598

 C12H3ClBr402                               530 - 538
 C12H2C128r402                              564,- 574
 C12HCl3Br402                               598 - 608
 C12Cl4Br402                                 634 - 640
                            A-72

-------
                 Table 15  (concluded)
Homolog                                    Mass range


C12H2ClBr302                               610 - 618
C12HCl28r502                               644 - 652
C12Cl3Br502                                678 - 638

C12HC1Br302                                688 - 698
C12Cl23r602                                722 - 732

C12ClBr702                                 764 - 776
                        A-73

-------
Table 16.  Limited Mass Scanning (IMS) Ranges for HOFs
 Homolog                                    Mass range
 C12H7C10                                   202 - 204
 C12HSC120                                  236 - 240
 C12H5C130                                  270 - 274
 C12H4C140                                  304 - 310
 C12H3C150                                  338 - 344
 C12HoCl60                                  372 - 378
 C12HC170                                   406 - 414
 C12C130                                    440 - 448

 C12H78rO                                   246 - 2*8
 C12H63r20                                  324 - 328
 Ct.,H5ar30                                  402 - 4Q8
 C12H43r40                                  480 - 488
 C12H3BrsO                                  558 - 566
 C12H28rsO                                  638 - 646
 C12HBr70                                   716 - 726
 C12Br30                                    796 - 806

 C12HsClBrO                                 280 - 28*
 C12H5Cl2BrO                                314 - 318
 C12H4Cl3BrO                                348 - 354
 C12H3Cl4BrO                                382 - 388
 C12H2Cl5BrO                                416 - 424
 C12HCls8rO                                 452 - 458
 C12Cl7BrO                                  484 - 494

 C12H5ClBr20                                358 - 364
 C12H4Cl2Br20                               392 - 398
 C12H3Cl38r20                               426 - 434
 C12H2Cl4Br20                               460 - 468
 C12HClsBr20                                494 - 502
 C12ClsBr20                                 528 - 538

 C12H4ClBr30                                436 - 442
 C12H3Cl2Br30                               470 - 478
 C12H2Cl3Br30                               504 - 512
 C12HCl48r30                                538 - 548
 C12Cl5Br30                                 572 - 582

 C12H3ClBr40                                514 - 522
 C12H2Cl2Br40                               548"-- 558
 C12HCl3Br40                                584 - 592
 C12Cl4Br40                                 618 - 626
                           A-74

-------
                 Table 16  (concluded)
Homolog                                    Mass range


C12H2ClBrsO                                594 - 600
C12HCl28r50                                628 - 637
C12Cl38r30                                 662 - 673

C12HC18rsO                                 672 - 537
Cl2Cl2Br60                                 706 - 715

C12ClBr70                                  750 - 760
                        A-75

-------
13.0  Quantitative Data Reduction

      The guidelines presented in this section are based on quantisation using
      surrogates and internal standards.   Isotope dilution techniques may also
      be applicable and may be substituted if validated prior tc use.  The QA
      record should show in detail  the alternative calculation used in arriving
      at final  concentration values.

      13.1  Once a chromatographic  peak has been identified as a HDD or HDF,
            the compound is quantitated based either on the integrated abun-
            dance of the SIM data or EICP for the primary characteristic ion
            in  Tables 13 and 14.   If interferences are observed for the
            primary ion, use the secondary and then the tertiary ion for quan-
            titation.  If interferences in the parent cluster prevent quanti-
            tation, an ion from a fragment cluster [e.g., M-63 (M-COC1) for
            PCDOs or PCDFs] may be  used.   Whichever ion is used, the RF should
            be  determined using that ion.   The same criteria should be applied
            to  the surrogate compounds.   If electron capture detection is used,
            the electronically integrated peak area is used for quantitation.

      13.2  Using the appropriate analyte-internal standard pair and response
            factor (RF.) as determined in Section 7.3, calculate the concen-
            tration of each peak using Equation 13-1.
Concentration (ng/g) =
                                           M
                                            is
                                   Ss  x Me  x RFh
                                                                     Eq.  13-1
            where:
         A.  = area of the characteristic ion (s) for the analyte
          n   HDD or HDF peak

        \.  = area of the characteristic ion (s) for the internal
              standard peak

              response factor of a given HOD or HDF congener

              mass of internal  standard added to extract (ng)

              mass of sample extracted (g)
                    RF,  =
                    Mis  =

                     M  =
      13.3   If  a  peak appears  to  contain  non-HDD  or  non-HDF  interferences,
            which cannot  be  circumvented  by  a  secondary  or-_lertiary  ion,
            either:

            13.3.1  Reanalyze  the sample  on  a  different  column  which  separates
                    the  interferences;  or

            13.3.2  Perform  additional  chemical cleanup  and  then  reanalyze
                    the  sample; or
                                    A-76

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            13.3.3   Quantitate  the  entire  peak  as  HDD  or  HDF.

            13.3.4   Reanalyze the sample using  a confirmation  technique [i.e.,
                    high  resolution mass spectrometry  (HRMS),  NCIMS,  etc.].

      13.4  Calculate  the recovery  of  the  surrogates using  the appropriate
            surrogate- internal  standard pair  and response factor (RFs)  as
            determined  in Section 7.5  using Equation 13-2.


                                   A  x M.  x 100

                                                                           '
            where:

                     A  = area of  the  characteristic  ion (s)  for the surrogate

                    A.  = area of  the  characteristic  ion (s)  for the internal
                     15    standard

                    RF  = response factor for  the  surrogate compound with
                      5    respect  to  the internal  standard (Eq.  7-1).

                    M.  = mass of  internal  standard added to  extract (ng)

                     M  = mass of  surrogate added  to  original  sample (ng)


      13.5  Correct the  concentration  of each  peak using Equation 13-3.   This
            is the final  reportable concentration.
            Corrected cone.  (ng/g)  =                                 *.  13-3
      13.6  Sum all  of the peaks  for each homo log,  and then sum those to yield
            the total  HDD and HDF concentrations  in the sample.   Report all
            numbers  in nanograms  per gram.   The  uncorrected concentrations.
            percent  recovery, and corrected concentration should be reported.

      13.7  Round off  all numbers reported to  two significant figures.


14.0  Confirmation

      If there is reason to question the qualitative identification (Section
      11.0), the analyst may choose to confirm that a peak is not a HDD or a
      HDF.  Any technique may be  chosen provided  that it is validated as
      having adequate  selectivity and sensitivity.   Some candidate techniques
      include alternate GC column (with EIMS detection), GC/CIMS, GC/NCIMS,
                                    A-77

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      high resolution EIMS, and MS/MS techniques.  Each laboratory  should
      validate confirmation techniques to show adequate selectivity between
      HDDs and HDFs and interferences and adequate sensitivity [limit of
      quantisation (LOQ)].

      If a peak is confirmed as being a non-HDD or a non-HDF, it may be
      deleted from the calculation (Section 13).   If a peak  is confirmed as
      containing both HDD or HOP and interferences, it should be quantitated
      according to Section 13.3.


15.0  Quality Assurance

      Each laboratory must develop a Quality Assurance Project Plan (QAPP)
      and submit that plan to the Agency for review and approval.  The QAPP
      documents how a laboratory intends to produce data which meet the data
      quality criteria specified in 40 CF3 707 and 766 (EPA 1987).

      A detailed QAPP submitted to the Agency will provide the necessary
      information required for the Agency to review the specific testing
      protocols for a commercial product.   Figure 5 provides a suggested
      table of contents for a QAPP as outlined in an EPA Office of Toxic
      Substances guide for preparation of QAPPs (EPA 1984).   The specific
      quality assurance objectives as stated in the final  test rule are sum-
      marized in Table 17.  Guidance for the development of a QAPP is also
      given in Appendix B.


16.0  Quality Control

      16.1  Each laboratory that uses these guidelines should operate a formal
            quality control (QC) program.   The minimum requirements of this
            program consist of an initial  demonstration of laboratory capabil-
            ity and the analysis of spiked samples as  a continuing check on
            performance.   The l-aboratory should maintain performance records
            to define the quality of data that are generated.  After a date
            specified by the Agency,  ongoing performance checks  should be
            compared with established performance criteria to determine if
            the results of analyses are within accuracy and precision limits
            expected of the method.  QC results should be  reported with the
            analytical results on a lot/batch basis.

      16.2  The analysts should certify that the  precision and accuracy of
            the analytical results are acceptable by the following:
                                                           •.
            16.2.1  The absolute precision of surrogate recovery, measured as
                    the RSD  of the integrated EIMS or ECD peak  area (A ) for
                    a set of samples, should be ± 30%.

            16.2.2  The mean recovery (R ) of at  least four replicates of a
                    quality control checR sample  should meet Agency-specified
                    accuracy and precision criteria.   This forms  the initial
                    data base for establishing control  limits (see  Section
                    16.3 below).

                                    A-78

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                              TABLE OF CONTENTS

1.0       Title page
2.0       Table of contents
3.0       Project description
4.0       Project organization and management
5.0       Personnel qualifications
6.0       Facilities, equipment, consumables, and services

          6.1  Facilities and equipment

               6.1.1  Evaluation
               6.1.2  Inspections and maintenance
               6.1.3  Calibration procedures and reference materials

          5.2  Consumables
          6.3  Services
7.0       Data generation

          7.1  Experimental design
          7.2  Sample collection
          7.3  Sample custody
          7.4  Laboratory analysis procedures
          7.5  Internal quality control checks
          7.6  Performance and system audits
3.0       Data processing
          8.1  Collection
          8.2  Validation
          8.3  Storage
          8.4  Transfer
          8.5  Reduction
          3.6  Analysis

9.0       Data quality assessment
          9.1  Precision
          9.2  Accuracy
          9.3  Representativeness
          9.4  Comparability
          9.5  Completeness
10.0      Corrective action
11.0      Documentation and reporting
          11.1  Documentation
          11.2  Quality assurance reports to management
12.0      References
Appendices
  Figure 5.   Table of contents for a quality assurance project plan (QAPP)
       as specified by the EPA Office of Toxic Substances (EPA 1984).
                                     A-79

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  Table 17   QC Procedures and Criteria for Analysis of Commercial Products
                  for 2,3,7,8-HDO and 2,3,7,8-HDF Congeners
Analysis event
        QC criteria
                                                        Corrective action
Instrument
mass calibration
Calibration
standards
 Samples
 QC samples
Must demonstrate accurate mass
calibration daily versus OFTPP
or PFK.

Calibration standards are used
to establish the working curve,
document response factors and
mass calibration.  After estab-
lishing the working curve, the
calibration standards are ana-
lyzed  to bracket sample analyses.
Response factors should vary
< 30%  from the working curve.

Precision of internal standards
must meet 50-150% recovery
criteria.

Precision of duplicate analyses
must meet ± 20%  criteria.

The  limit of quantisation  (LOQ)
must meet 0.1 ppb for 2,3,7,8-
HDD  congeners and 1.0 ppb  for
2,3,7,8-HDF congeners.   LOQ  =
10  times background  signal-to-
noise.  Limit of detection (LOD)
must be calculated.   LOD  = 3 times
background  signal-to-noise.

Method blanks  analyzed  with  each
 sample batch.   Spiked  samples
 50-150% recovery.
Recalibration versus
OFTPP or PFK.
If RF values vary by
more than ± 30%,
sample analyses must
be repeated following
recalibration.
If accuracy, preci-
sion, or sensitivity
requirements are not
met, the sample must
be reanalyzed.
 If method blank gives
 any positive response,
 repeat analysis.   If
 recovery results do
 not meet the criteria,
 the sample must be
 reanalyzed.
                                       A-80

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16.3  Control  limits - The laboratory should establish control limits
      using the following equations:

              Upper control  limit (UCL) = RC = 3 RSDC

              Upper warning  limit (UWL) = RC + 2 RSOC

              Lower warning  limit (LWL) = RC - 2 RSDC

              Lower control  limit (LCL) = RC - 3 RSDC

      These may be plotted on control charts.   If an analysis of a check
      sample falls outside the warning limits, the analyst shouia be
      alerted  that potential  problems may exist and corrective act-Ion
      may be indicated.   If  the results for a check samole fall  outside
      the control  limits, the laboratory should take corrective  action
      and recertify the performance of the method (Section 16.2) before
      proceeding with analyses.   The warning and control  limits  snould
      be continuously updated as check sample replicates  are added to
      the data base.

16.4  Before processing any  samples,  the analyst should demonstrate
      through  the analysis of a reagent blank that all glassware and
      reagent  interferences  are under control.  Each time a set  of sam-
      ples is  analyzed or there is a change in reagents,  a laboratory
      reagent  blank should be processed as a safeguard against loss of
      data caused by contamination.

16.5  Procedural QC - The various steps of the analytical procedure
      should have quality control measures.   These include but are not
      limited  to:

      16.5.1  GC performance  - See Section 7.1 for performance criteria.

      16.5.2  MS performance  - See Section 7.2 for performance criteria.

      16.5.3  Quantisation -  At least 10% of all  manual calculations,
              including peak  area calculations, should be checked.
              After changes  in computer quantisation routines, the
              results should  be manually checked.

16.6  A minimum of 10% of all samples, one sample per month or one
      sample per matrix type, whichever is greater,  selected at  random,
      should be run in duplicate to monitor the precision of the method.
      A difference of 20% or  less should be achieved.   If the difference
      is greater than 20%, the analyst should investtgate the cause of
      the imprecision and reanalyze the samples.   Failure to satisfy
      this requirement could  invalidate the results  obtained for samples
      analyzed since the last precision determination.
                              A-81

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      16.7  A minimum of 10% of all  samples, one sample per mor.th, or one
            sample per matrix type,  whichever is greater, selected at random,
            should be analyzed by the standard addition technique.  Two ali-
            quots of the sample are  analyzed, one "as is" and one spiked with
            an appropriate amount of HDD and/or HOP congeners.   The samples
            are analyzed together and the quantitative results  calculated.
            The recovery of the spiked compounds (calculated by difference)
            should be 50-150%.   If the sample is known to contain specific
            HDD or HDF isomers, these isomers should be used for spiking.
            If the aporoximate concentrations of analytes are known, the
            amount added should be such that the spiking level  is 1.5 to 
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                                    A-88

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                         APPENDIX 3
QUALITY ASSURANCE PROJECT PLAN FOR MEASUREMENT OF HALOGENATED
      DIBENZO-a-OIOXINS (HMDs) AND OI3ENZQFURANS (KDFs)
                         Prepared by

                       Dr.  John Smith
               Environmental Protection Agency
                 Office of Toxic Substances
                 Chemical Regulation Branch
                       Washington, DC
                             B-l

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        QUALITY ASSURANCE PROJECT PLAN FOR MEASUREMENT OF HALOGENATED
              DIBENZO-o-OIOXINS (HMDs) AND DIBENZOFURANS (HDFs)


          For quality assurance (QA) to be an effective method of measurement
validation of qualification,  a  Quality Assurance  Project Plan  (QAPP)  must be
prepared and should include the following:  history, management, and ultimate
location of samples; sampling and sample collection procedures; and extraction,
instrumental analysis, and data reduction procedures.  The QAPP documents how
a laboratory intends to demonstrate its capability to produce data of accept-
able quality.   The proposed rule is concerned with the following four classes
of compounds:   brominated  dibenzofurans,  brominated dibenzodioxins, chlori-
nated dibenzofurans,  and  chlorinated  dibenzodioxins.   Of specific  interest
within these classes  are  the compounds substituted with from  four  to  seven
chlorine and/or bromine atoms per molecule.   Hereafter these compounds will
collectively be referred  to  as  halogenated dibenzofurans (HDFs) and haloge-
nated dibenzodioxins (HDDs).

          An accurate trace  or  history of the  life of a samole  to  be  chem-
ically analyzed for HOFs and HOOs must be assembled and should contain infor-
mation beginning with a  description of the system, scheme,  or survey design
for sample collection.  The  sample collection  scheme must be  statistically
designed to allow  measurement of variance of the  sampling site.  A written
record,  sometimes  called a traceaoility form, shall  follow the  sample.-  Nec-
essary contents of the record are written general  descriptions of what happens
to the sample,  schedules and timetables,  disposition, and handling.   Following
final chemical  analysis, the  last  entry in this trace should be  the ultimate
location of the sample.   Any further use (particularly for another activity),
movement, or examination of the sample should be added to the history.

          Details   of  the  sampling or sample collection  procedures  are the
second section  of the QAPP.   Since the history describes  handling disposition,
schedules,  and  general descriptions of what happens in sampling, the require-
ment here is a detailed description of sampling and sample collection.  Rea-
sons for using  a specific or general sample selection process and reasons  for
not using other processes must be included here.  The rationale for any modi-
fications made to  a method must also be  given.   Estimates  of how well the
selected samples represent the  material to be characterized are  an  essential
part of  quality assurance.   The greater the variability  of composition of a
material, the  more frequently sampling is required for estimation and charac-
terization purposes.  In determining or estimating errors generated in sample
collection,  control samples or  blanks and HDF and  HOD reinforced controls or
standards (native  compounds  or, preferably,  isotopically labeled compounds)
must be  sent to  the collection site(s) and  returned  with  the samples for
identical handling  and  treatment.   Duplicate samples, which are collected,
documented,  and handled the same as other samples, are necessary for recovery,
precision,  and  accuracy determinations.

          The  third section  of  the QAPP is  a description of the extraction
and chemical analysis or screening test procedures.  The  QA record should  show
in detail the  mathematical  calculations performed in arriving at final  concen-
tration values.   To determine both extraction efficiency  and measurement effi-
ciency,   it  is necessary to use  chemical ^tandards  of HDF and HDD mixtures or


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individual HOP and HOD compounds in control samples.  Once capabilities  have
been established, the operator must determine precision and accuracy  and use
those determinations to designate acceptable bounds for analytical performance.
The operator must keep control chart records to ensure that instrument  read-
ings of standards fall  within the range of acceptable performance.  The oper-
ator must establish and describe the quantitative  range of an  instrument and
analytical procedure.   Specific requirements are as follows:

               For chemical  analysis,  these procedures must be demonstrated
               capable to  reproducibly and repeatedly quantitate HDDs and
               HDFs at the  required limits  of quantitation.  Quality  control
               check  sample analysis   begins  the   determination  of system
               capabilities.

               For HDDs,  at least two  analyses of the same spiked standard at
               the required  LOQ  must  be quantifiable to within ± 20% of the
               true value.   The  recovery  of standard spiked into a product
               material at  the required LOQ and run  through the  entire  chem-
               ical  analysis  must  be  within 50-150% of  the  amount spiked,
               with reference  to the  instrument standard, 99% of the time.

               For HOFs,  at least two  analyses-of the same spiked standard at
               the required  LOQ  must  be quantifiable to within ± 20% of the
               true value.   The  recovery  of standard spiked into a product
               material at the required LOQ and run through the entire chemical
               analysis must be  within 50-150% of  the  amount  spiked, with
               reference to the instrument standard, 99% of the time.

          Qualitative requirements  include:   response  factors  for HDFs  and
HDDs to  be measured,  instrument  hardware  and operating conditions (including
type and  source  of  columns,  carrier gas and flow rate, operating temperature
range, and ion source  temperature), and structural  assignment  and/or  library
matching of mass spectra.   For both qualitative and quantitative measurements,
the  instrument operator  should be  blind to the nature or source of samples,
particularly to  duplicates,  blanks,  and brominated and spiked samples.  The
limit  of  detection  (LOD)  and limit of quantitation (LOQ) shall be described
for  each  material.   Tentative LOD and  LOQ  definitions  are equal  to  three
times  and ten times background noise,  respectively.  Details of  quantitative
calibration procedures for the known and/or expected range of the HDF and  HDD
levels in actual  samples complete this section of the QAPP.

          The last section of the QAPP must include the results of laboratory
participation in  round robin analytical programs,  the  results  of performance
audits, the results of systems audits, analytical  results of performance audit
samples,  persons  responsible for all aspects of sampling,  chemical analysis,
data  analysis,  corrective  actions, and quality assurance/equality control
This section also must include a description of how  problems are handled and
documented, and  how corrections  in working  level notebooks are indicated and
explained.
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                        APPENDIX C
GUIDANCE FOR SAMPLING HALOGENATED DIBENZO-D.-DIOXINS (HDPs)
                 AND DIBENZOFURANS (HDFs)
                        Prepared by

                      Dr.  John Smith
              Environmental Protection Agency
                Office of Toxic Substances
                Chemical Regulation Branch
                      Washington, OC
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         GUIDANCE FOR SAMPLING HMJ.OGENATED DIBENZO-&-DIOXINS (HDDs)
                          AND DIBENZQFURANS (HDFs)


          The following are  recommended  steps  for collecting samples of the
following four classes  of  compounds:   brominated dibenzofurans,  brominated
dibenzodioxins,  chlorinated  dibenzofurans, and chlorinated  dibenzodioxins.
Of specific  interest within  these classes  are  the compounds  substituted with
from four to  seven  chlorine  and/or bromine atoms per  molecule.   Hereafter
these compounds  will collectively be referred to as  halogenated dibenzofurans
(HDFs) and halogenated  dibenzodioxins  (HDDs).   Specific  subgroups within the
collective group adaressed by the regulation will be more specifically named.
All of the following steps should be carefully recorded.

          1.    Determine the  objectives of the samel ing exercise.

          In the case of this proposed rule, some objectives might  be  (1)  to
measure HDF and  HDD levels occurring as oy-products  or impurities in a product
to determine compliance with the rule, and (2)  to  know the limitations of
these measurements.

          2.    Describe the system or production process from which the sample
               materials will be collected.

          The-samples collected can only provide information about the product
from which they  have been collected, and the completeness of this information
depends on how,  at what time(s), where, and for what duration the samples are
collected.  Of particular  importance  in  determining a sampling plan for the
process are  the  continuity,  homogeneity,  and  cyclic characteristics of the
production process.   Compositing of samples for chemical analysis has benefits
in reducing  analytical  costs  but  risks in  requiring lower  instrument quanti-
fication  levels and reducing the ability  to identify  whether HDFs  and HDDs
are from uniform concentrations in all members of a composite or high concen-
trations in one  member  and no concentrations in the other members of the com-
posite.  Each of the seven samples described in the "Guidance for a Sequential
Approach to the  Sampling of Dibenzofurans and Dibenzodioxins" may be a randomly
selected  composite,  but the  seven may not be composited unless the level of
concern is revised  to one-seventh of the currently proposed  levels.

          3.    Prepare  for sample collection operations.

          Professional  judgment  must  be assisted  by proven sampling methods.
To make  a complete  analysis, sampling should  also determine the presence or
absence of HDFs  and HDDs in locations or situations not necessarily judged to
be most important.

          The proposed  rule requires random sampling.  A random sample selec-
tion  system  includes  all  possibilities and systematically  chooses from among
a  list or compilation of those possibilities.   The way of choosing  from among
the possibilities is the use of a random number table  or through an automated
random number generator.   A  haphazard selection system,  the result  of  selec-
tion  through  convenience  and/or purposeful neglect, includes only  a portion
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of all possibilities,  has  very littla estimating power,  and  should not be
considered a random selection procedure.

          For the purpose  of incorporating this sampling plan with  the  EPA
"Guidance for a Sequential  Approach to the Sampling of Brominated and Chlori-
nated Dibenzofurans and  Dibenzodioxins,"  the sampling unit for each process
will be a  cycle.   A cycle  is  defined  as  the time interval representing a
period within which the  activity mix of  a process is essentially  repeated.
This cycle may be a batch,  a shift, a day, or some other time period.

          It is  important  to use random selection procedures in determining
the sampling time  and  location for each  individual  cycle (the details  for
these determinations are given below in parts  b  and c).   Cycles should be
sampled,  one at  a  time,  until as  many  as  seven cycles  have  been  sampled.
If two samples  from any  of  the  seven or  fewer  cycles are analyzed  and  show
measurement  levels  of  HOFs and HDDs above the  limit of quantitation,  the
material  is  not  in  compliance with the rule.   If  none  of the seven cycles
show HDDs  above  the required  LOQ, no  further  sampling  is necessary  until
there are changes in the  process as described below.

          If there are changes known or suspected to change the generation of
HDFs and HDDs as by-products or impurities in a process  or a cycle, the  seven-
cycle sampling exercise  must be done again.   For  each  defined process  and
independent cycle:

               a.  The size  of a  sample of a material to  be analyzed  will be
determined by the  kind of  material to be sampled and the sensitivity of the
analytical  method.  If the material  is gaseous, a sample size of 10 m3 is
usually sufficient  for analysis.   If the material is aqueous or liquid, a
sample size  of 1 L  is  usually  sufficient  for analysis.  If the material  is a
solid, a sample size of 100 g is usually sufficient for  analysis.  Any sample
size must  have proper  documentation demonstrating capability  to  measure  HDDs
at the required limits of quantisation.

               b.  Location(s)  of sample  collection  are defined as a subset
of all possible locations of storage of a given material.   Physically possible
sample collection  sites  are  listed  and  chosen  through random  selection
procedures.

               c.  In  processes  where  over seven batches of  a material  are
produced in  a year,  sample collection times must be  selected from  a  random
time period within each individual batch production cycle.  The cycle time is
completely segmented into  intervals  long  enough to collect a  sample,  and one
interval  is randomly selected for the actual  collection.  For processes where
under seven  batches of material  are produced, random  selection of  seven  sam-
ples from  a  list of production time segments  for  all batches is employed.

               d.  Quality  assurance of the sampling and actual sample collec-
tion exercise includes documentation of:   the sampling plan,  sample collection
procedures, and any traceability records.   Special notation should be made of
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any deviations from original  plans  and projections recorded at  an earlier
stage of the sampling  exercise.   Especially:

                    dates and times  of sample collection and chemical  analysis
                    of samples;

                    exact location and time of sample  collection;

                    process  or  product batch  or  lot  identification:  and

                    traceability  records employed  in  the sample collection
                    and ultimate location  of the samples.
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                           APPENDIX 0
GUIDANCE FOR A SEQUENTIAL APPROACH TO THE SAMPLING OF HALQGENATED
        DIBENZO-a-QIOXINS (HDDs) AND DIBENZOFURANS (HDFs)
                           Prepared by

                        . Or.  John Smith
                 Environmental Protection Agency
                   Office of Toxic Substances
                   Chemical  Regulation Branch
                         Washington, DC
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       GUIDANCE FOR A SEQUENTIAL ABROACH TO THE SAMPLING OF  HALQGENATED
               DIBENZO-a-OlOXINS CHDDs) AND DIBENZOFURANS (HDFsT	


 INTRODUCTION

           This is the  sequential  sampling  scheme  referred  to in  the  Proposed
 Rule  for  Processing,  Distribution, and Use of  Certain Designated Chemical
 Substances Suspected to Produce and/or Be Contaminated with Dioxins and Furans
 (49 FR 28172, 28182, July 10, 1984).

           For this  proposed rule, dioxins and  furans  are  in realitv  four
 classes of compounds:   brominated dibenzofurans,  brominated dibenzodioxins,
 chlorinated dibenzofurans,  and chlorinated  dibenzodioxins,  and specifically
 within these  classes  the compounds  substituted with  from four to  seven
 chlorine  and/or bromine atoms per molecule.  Hereafter these particular com-
 pounds will collectively  be referred  to  as halogenated dibenzofurans (HDFs)
 and halogenated  dibenzodioxins  (HDDs).   Specific subgroups  within the collec-
 tive group addressed  by  the  regulation  will  be more  specifically  named.

           This document contains a statement of  the sequential  decision scheme
 the rationale for its  use,  the details of  sample  selection (how  much,  when,
 and where), and  the  statistical properties  of  the  sequential  decision scheme
 ihis scheme is part of  an overall  activity  to provide  guidance to  those who
 are in a position  to  respond to the proposed regulations.


 PROPOSED TRUNCATED SEQUENTIAL DECISION SCHEME

           For  a  given material  at  the production/formulation site, select a
 sample  and measure the concentration of HDFs and HDDs.  The  sampling scheme
 must be  used  for all tests  and analytical  evaluations  for  determination  of
 HDFs  and HDDs.  A measurement may  be obtained by a  screening method which has
 been  demonstrated  by the tester to a  documented degree of sensitivity and
 specificity.   The  confirmatory  method for identifying the HDFs and HDDs and
 the  concentrations of  these  compounds  is  high  resolution gas  chromatography/
 high  resolution mass spectrometry  (HRGC/HRMS).

          The  product  sample selection  procedure is repeated until either a
 product sample is found to contain HDFs and/or HDDs or seven samples have be°n
 processed,  in which case declare the production of the sampled material  to be
 free  from HDFs and HDDs.  If any of the concentrations of HDDs and/or HDFs in
 a single product sample  exceeds the required LOQ, the product will be deemed
 to  contain  HDDs and/or  HDFs.  In HRGC/HRMS a resolvable chromatograohic peak
 is considered a single brominated  dibenzofuran  compound, a  single chlorinated
dibenzofuran compound, a single brominated dibenzodioxin compound, or a  single
chlorinated dibenzodioxin  compound.   The  number  of samples, N, required is
not to exceed seven.
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REASON FOR NOT USING A SINGLE SAMPLE TO DETERMINE COMPLIANCE

          Analysis of a  material  for HDFs and HDDs  being generated as by-
products or impurities  is  subject to two main souces of variation:  samples
may vary over time and/or  location at  the site.  Secondly,  the measured  con-
centration of HDFs  and  HDDs will vary  among  repeated measurements on the
same sample.   Therefore, the  measured concentration of a particular single
screening test or single gas  chromatographic peak will vary among samples
selected randomly in  time  and space so  that  a  single sample would not  be
representative of the underlying process except for the extreme cases.

          In  the  following paragraphs the variable  nature of ooservations
will be expressed by the symool  e,  the probability that a sample will  have a
measured concentration below  the  required LOQ in a particular gas chromato-
graphic peak.


JUSTIFICATION FOR USING  SEQUENTIAL SAMPLING

          Given  the analytical costs associated with HRGC/HRMS,  it is  impor-
tant to  utilize  statistical  procedures which minimize the number of samples
which need to be analyzed.   Sequential monitoring of a material  allows a  de-
termination of the presence of HDFs  and/or HDDs to be based on fewer samoles,
on  the  average, than would be  required under  an equivalent  fixed  sample  size
decision rule because of the  large  number of samples required to describe a
production having undemonstrated variability of HOF and HDD content.  For eacn
such screening test or HRGC/HRMS analysis, this efficiency increases with the
number of potential  HDFs and HDDs in the material  and decreases as the magni-
tude of  e  increases.  For materials containing sufficiently high concentra-
tions of even a  single  HDF or HDD compound (i.e.,  e  is  close  to  zero  for
its associated screening test or peak), only one sample will be required to
declare the presence of  HDFs  or HDDs.  For most multiple  congener  materials,
the use  of the proposed  scheme would probably result in the  analysis of  only
four or five  samples to  identify materials containing HDDs and HDFs above the
required LOQ.   Even for  materials containing a single congener at levels  above
the required LOQ, the application of the  scheme would probably result  in  the
analysis of fewer than seven samples to declare the presence of HDFs and  HDDs
when  e  is no larger than  0.7.


TIMES. LOCATIONS.  AND QUANTITIES

          A material may be  produced through a group of  activities, within
one or  more  locations at a site.   The production of the material is assumed
to be cyclic  in  nature,  and within each cycle the distribution and concentra-
tion ranges of brominated  and chlorinated dibenzofurans  and  dibenzodioxins
are assumed to be consistent.   Each  cycle, then,  is a time interval represent-
ing a period within which  the  activity mix of the production  of  the material
is essentially repeated,  such as a batch, shift,  or day.

          Samples under  the proposed sequential  sampling plan will be  selected
from separate cycles of  the production process.   Further description of sample
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 selection appears  in "Guidance for Sampling Brominated and Chlorinated Dibenzo-
 furans  and Dibenzodioxins" (Appendix C).  It is important to randomly select
 the sampling time and location independently fop each cycle.  Any deviations
 from  this random  selection must be documented  and  justified.   As  many as seven
 cycles  will  be  required  under  the  proposed  procedure.   Changes  in the produc-
 tion  process,  especially changes judged to potentially affect brominated and
 chlorinated  dibenzofuran and dibenzodioxin  levels,  require  that, the entire
 sequential  testing effort be  redone.   If a  single  production  process is the
 potential  source of  brominated and chlorinated  dibenzofuran  and dibenzodioxin
 exposure  through  several media, each medium should be separately tested by
 the sequential  sampling.

           The size of the sample  to  be chemically analyzed depends  on  the
 medium  of the  sample and  the  intended  analytical  method.   Sample sizes  are
 recommended  in  the "Guidance for Sampling Brominated and Chlorinated  Oibenzo-
 furans  and Dibenzodioxins."


 STATISTICAL PROPERTIES OF  THE  PROPOSED  DECISION SCHEME

           In general,  two  decision errors are  possible:   (1)  declaring mate-
 rials in compliance to be  not  in compliance, and (2) declaring processes which
 are not in compliance to  be in  compliance.   Choice of 0.1 ppb as the test
 threshold  and requiring  two samples to exceed it eliminates any  significant
 likelihood of committing an error of the first type.  The probability of com-
mitting an error  of  the second type  decreases with the number of potential
brominated and  chlorinated dibenzofuran and dibenzodioxin congeners present
 in the process  and the maximum number of required samples, and increases.with
the magnitude of  e  for each such peak.  The proposed maximum number of seven
samples was chosen because it  results  in an acceptable  probability of the
second  error without  the requirement  of an excessive amount of samples to be
analyzed to determine the  presence  of HDFs and HDDs.   For  the seven samples,
the error  of the  second type will  be less  than 0.25 for a typical process '
(i.e., one having five or more  peaks) provided the   e  values for the process
are all no larger  than 0.92 (assuming unconditional  independence  or complete
gas chromatographic  resolution of individual HDF and  HDD  concentrations).
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