United States
Environmental Protection
Agency
Industrial
Laboratory
Cincinnati OH 4S2W
EPA-600/2-80-157
June 1980
Research and Development
Dioxins

Volume  II.
Analytical Method for
Industrial Wastes

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                 RESEARCH REPORTING SERIES

 Research reports of the Office of Research and Development, U.S. Environmental
 Protection Agency, have been grouped into nine series. These nine broad cate-
 gories were established to facilitate further development and application of en-
 vironmental technology.  Elimination of traditional grouping  was  consciously
 planned to foster technology transfer and a maximum interface in related fields.
 The nine series are:

       1.  Environmental Health Effects Research
       2.  Environmental Protection Technology
       3.  Ecological Research
       4.  Environmental Monitoring
       5.  Socioeconomic Environmental Studies
       6.  Scientific and Technical Assessment Reports (STAR)
       7.  Interagency Energy-Environment Research and Development
      8.  "Special" Reports
      9.  Miscellaneous Reports

 This report has been assigned to the ENVIRONMENTAL MONITORING series,
 This series describes research conducted to develop new or improved methods
 and instrumentation for the  identification  and quantification  of environmental
 pollutants at the lowest conceivably significant concentrations.  It also includes
 studies to determine the ambient concentrations of pollutants in the environment
 and/or the variance of pollutants as a function of time or meteorological factors.
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia  22161.

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                                              EPA-600/2-80-157
                                              June 1980
                    DIOXINS:
                   VOLUME II.
     ANALYTICAL METHOD FOR INDUSTRIAL WASTES
                       by

     T. 0. Tiernan, M. L. Taylor, S. D.  Erk,
     0. 6. Solch, G. Van Ness, and J. Dryden
The Brehm Laboratory and Department of Chemistry
             Wright State University
               Dayton, Ohio  45435
            Contract No. 68-03-2659
                 Project Officer

                David R. Watkins
     Industrial Pollution Control Division
 Industrial Environmental Research Laboratory
             Cincinnati, Ohio  45268
  INDUSTRIAL ENVIRONMENTAL RESEARCH LABORATORY
        OFFICE OF RESEARCH AND DEVELOPMENT
      U.S.  ENVIRONMENTAL PROTECTION AGENCY
             CINCINNATI, OHIO  45268

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                                 DISCLAIMER


     This report  has  been reviewed by  the  Industrial  Environmental  Research
Laboratory,   U.S.   Environmental   Protection   Agency,   and   approved  for
publication.  Approval does not signify that the contents necessarily reflect
the views and  policies of the U.S. Environmental Protection Agency,  nor does
mention  of  trade names  or  commercial  products  constitute endorsement  or
recommendation for use.
                                     n

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                                   FOREWORD
     When energy and  material  resources are extracted,  processed,  converted,
and used, the  related pollutional  impacts on our environment and even on  our
health often require  that new and  increasingly more efficient pollution con-
trol  methods  be  used.    The  Industrial  Environmental  Research  Laboratory-
Cincinnati  (lERL-Ci)  assists  in developing  and  demonstrating  new  and  im-
proved  methodologies  that  will  meet  these  needs  both  efficiently  and
economically.

     This report  is one  of a three-volume  series dealing  with a group  of
hazardous chemical  compounds  known  as  dioxins.  The extreme  toxicity of  one
of  these  chemicals, 2,3,7,8-tetrachlorodibenzo-p-dioxin  (2,3,7,8-TCDD),  has
been a concern  of  both scientific  researchers and the  public for many years.
The sheer mass  of  published information that  has  resulted from  this  concern
has  created difficulties  in  assessing  the  overall   scope  of the  dioxin
problem.    In  this  report  series  the  voluminous  data  on  2,3,7,8-TCDD  and
other dioxins are  summarized  and assembled in  a  manner that allows compari-
son of related  observations from many sources; thus,  the series serves as a
comprehensive guide in evaluation  of  the environmental  hazards of dioxins.

     Volume I  is  a state-of-the-art review of dioxin  literature.   Detailed
information is  presented  on the chemistry, sources, degradation,  transport,
disposal, and   health  effects of  dioxins.   Accounts  of  public  and  occupa-
tional exposure to  dioxins are also included.  Volume  II details the devel-
opment of a new analytical method for  detecting  part-per-trill ion levels of
dioxins  in  industrial wastes.   It  also includes a review  of the  analytical
literature on methods  of  detecting dioxins in various  types of environmental
samples.   Volume  III  identifies various  routes of  formation of  dioxins in
addition  to  the  classical route  of the  hydrolysis  of  chlorophenols.   The
possible  presence  of dioxins  in basic organic chemicals  and pesticides is
addressed,  and production locations   for  these  materials  are identified.

     For  further   information,   contact Project  Officer  David  R.  Watkins,
Organic  and Inorganic  Chemicals  Branch,  lERL-Ci.   Phone  (513)  684-4481.


                                             David G. Stephan
                                                 Director
                               Industrial  Environmental  Research  Laboratory
                                                Cincinnati
                                     m

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                                    PREFACE
      This report  is  Volume II  in a series  of  three  reports dealing with a
 group of  hazardous  chemical compounds  known as dioxins.   This  volume dis-
 cusses the development of  a new  analytical technique for identifying dioxins
 in  industrial   wastes,   and presents  a  bibliography  of   other  analytical
 methods  for determining  dioxins   in  various  types  of environmental samples.
 Other volumes  of this series  examine  the occurrence,  environmental  trans-
 port,  toxicity,  and  disposal   of  this  class  of  compounds, the  detailed
 chemistry of dioxin  formation,  and  the  commercial products with potential
 for  containing  dioxin  contaminants.

      An  extensive body of  published  literature  has appeared during the past
 25 years  that  has been concerned primarily  with one  extremely toxic member
 of  this   class   of  compounds,   2,3,7,8-tetrachlorodibenzo-p-dioxin.   Often
 described in  both  popular and   technical  literature  as  "TCDD"   or  simply
 "dioxin,"  this  compound  is one of the  most  toxic  substances  known  to
 science.   This   report  series  is  concerned not  only with  this compound,  but
 also  with all  of its  chemical  relatives  that  contain the  dioxin nucleus.
 Throughout these reports,  the  term  "TCDD's"  is used  to indicate  the family
 of 22 tetrachlorodibenzo-p-dioxin isomers, whereas the  term  "dioxin" is used
 to indicate  any compound  with  the  basic dioxin  nucleus.    The  most  toxic
 isomer  among those  that  have  been  assessed is specifically  designated  as
 "2,3,7,8-TCDD."

     The  objective  in  the  use  of these  terms is to clarify  a point of tech-
 nical  confusion  that  has  occasionally  hindered  comparison  of  information
 from  various  sources.   In  particular,  early  laboratory   analyses  often
 reported   the  presence  of "TCDD,"  which   may  have   been   the  most-toxic
 2,3,7,8-isomer  or may  have been a  mixture  of several of  the  tetrachloro
 isomers,  some   of which  are relatively  nontoxic.    Throughout this  report
 series, the  specific  term 2,3,7,8-TCDD is used when it was the intent of the
 investigator to  refer  to  this  most-toxic  isomer.  Since  early  analytical
methods  could  not  dependably  isolate  specific  isomers from environmental
samples,  the generic  term "TCDD's" is used when this term appears  to be most
appropriate in  light of present technology.
                                     IV

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                                  ABSTRACT
     The overall  objective  of this research project was to develop a unified
analytical  approach  for  use  in  quantifying  part-per-trill ion  levels  of
tetrachlorodibenzo-p-dioxins (TCDD's) in various chemical wastes.

     The  EPA  provided Brehm  Laboratory of Wright  State University  with  17
waste  samples  from plants  manufacturing  trichlorophenol,  pentachlorophenol,
and hexachlorophene, and from plants processing wood preservatives.

     The  extraction procedure  developed  for isolating  the TCDD's  from  the
various  types  of sample  matrices is fully  described.   Analysis  was accom-
plished  using  highly  specific and sensitive coupled gas chromatographic-mass
spectrometric  (GC-MS)  methods.    Both  low and high  resolution  MS  techniques
were  employed.    This  methodology  is  also  described   in  detail.    The  pro-
cedures  presented  in  this report were acceptable for most of the  industrial
process  samples  provided.   TCDD's were  detected  and  quantitatively deter-
mined  in several of  the samples  at levels  in  the ppt  to ppm range.   One
sample,  identified as a  trichlorophenol  stillbottom,  was found  to contain
40 ppm TCDD's.   This  method was not applicable for  wood or woodlike products
and difficulties  were also  encountered with some samples  that were suscep-
tible to emulsion formation in the preparation stages.

     The Brehm  Laboratory submitted this report in  fulfillment of a subcon-
tractual   effort  with  Battelle  Columbus  Laboratories,   supported  through  a
prime contract  between Battelle and the U.S.  Environmental Protection Agency
(Contract No.  68-03-2659).   This report covers the  period October  1, 1978 to
March 31, 1979 and work was completed as of March 3, 1979.

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                                  CONTENTS
Foreword
Preface
Abstract
Figures
Tables
List of Abbreviations
Acknowledgment
I.   Introduction
2.   Analytical Background
3.   Analytical Method
4.   Discussion and Results
5.   Conclusions and Recommendations
References
Appendix A.    Principles of GC-MS
Appendix B.    Other Instrumental Methods
Appendix C.    Literature Review
Page

 i i i
  iv
   v
viii
  ix
   x
  xi
   I
   4
   8
  13
  43
  45
  49
  57
  60
                                     vii

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                                   FIGURES
Number                                                                Page

  1       Mass chrotnatogram of extract of EPA sample 2 at m/e 322
          obtained with GC-QMS.                                         16

  2       High pressure liquid chromatogram of sample 2.                23

  3       High pressure liquid chromatogram of 2,3,7,8-TCDD standard.   24

  4       Four-ion mass fragmentogram of benzene solvent  blank
          obtained with GC-MS-30.                                       26

  5       Four-ion mass fragmentogram of 50 pg of 2,3,7,8-TCDD
          and 1 ng 37Cl4-2,3,7,8-TCDD obtained with  GC-MS-30.           27

  6       Four-ion mass fragmentogram of sample 12700 obtained
          with GC-MS-30.                                                28

  7       Four-ion mass fragmentogram of sample 5 obtained
          with GC-MS-30.                                                29

  8       Dual-ion mass fragmentogram of sample 2, obtained
          with GC-MS-30,  mass  resolution 1:12,500.                      35

  9       Dual-ion mass fragmentogram of 150 pg of 2,3,7,8-TCDD
          standard obtained with GC-MS-30,  mass resolution 1:12,500.    36

10       Mass  fragmentograms  using  GC-MS-30 of mixtures  of
          2,3,7,8-TCDD  with other  chlorinated compounds.                38

11       Mass  spectrum of  2,3,7,8-TCDD  standard and sample 2
          (mass range m/e 330  to m/e 250).                              41

12       Mass  spectrum of  2,3,7,8-TCDD  standard and sample 2
          (mass range m/e 250  to m/e 150).                              42
                                  viii

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                                   TABLES
Number                                                                Page

  1       Samples Used in Development of Analytical  Method for
          TCDD's in Industrial  Wastes                                  14

  2       Elution of TCDD's in  Extracts of Sample 2                     17

  3       Content of TCDD's in  Column Fractions for  Sample 2           19

  4       Elution of TCDD's in  Extracts of Sample C04131               20

  5       Results of GC-MS-30 Analyses of EPA Samples for TCDD's       25

  6       TCDD Isomer Content of Column Fractions of 2,3,7,8-TCDD
          Spiked Samples                                               31

  7       Recoveries of 2,3,7,8-TCDD Spiked Samples  Following
          Alumina Column Chromatography                                33

  8       Results of GC-MS-30 Analyses of 37Cl4-2,3,7,8-TCDD
          Spiked Samples                                 .             34

  9       Relative Intensities  of Major Ions Observed in Mass
          Spectral Scans                                               40
                                     ix

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                           LIST OF  ABBREVIATIONS
   cm               centimeter
   DDE              2,2-bis-(p-chlorophenyl)-l,l-dichloroethylene
  GC-EC             gas  chromatography-electron  capture
   eV               electron  volt
   9                gram
   GC               gas  chromatography
  GC-MS             gas  chromatography-mass spectrometry
GC-MS-30           gas  chromatography-mass spectrometry (high resolution)
  GC-QMS            gas  chromatography-quadrupole mass spectrometry (low
                     resolution)
   HPLC             high-pressure  liquid chromatography
   I.D.             inside diameter
   kg              kilogram
   LD50             lethal dose to 50% of test group
   m               meter
   m/e              mass to charge ratio
   ml              milliliter
  ml/min            milliliter/minute
   mm              millimeter
   MS              mass spectrometry
   ng              nanogram
   PCP             pentachlorophenol
   PCB             polychlorinated biphenyl
   pg              picogram
   ppb             parts per billion (ug/1 or ng/ml)
   ppm             parts per million (mg/1 or ug/ml)
   ppt             parts per trillion (ng/1 or pg/ml)
  PSIG             pounds per square inch gage
2,4,5-TCP          2,4,5-trichlorophenol
 TCDD's            tetrachlorinated dibenzo-p-dioxins; 22 possible isomers
   ug             microgram
   UV              ultraviolet
   V              volt

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                              ACKNOWLEDGMENT
     This  report  was prepared  for the U.S.  Environmental  Protection Agency
by the  Brehm  Laboratory and Chemistry Department of Wright State University,
Dayton, Ohio.   Dr.  T.O.  Tiernan was the Principal Investigator with Dr.  M. L.
Taylor  and  S.D.  Erk as Co-Principal Investigators.   Mr.  Dave Watkins was the
Project Officer for the U.S. Environmental Protection Agency.

     A  review of the  analytical  literature  for determination of  TCDD's in
various  sample  matrices   was   compiled  by  Battelle Columbus  Laboratories,
Columbus, Ohio,  and constitutes a significant addition to this report.  This
material was used to develop Appendices B and C.

     Final  compilation  of  this  report for  integration  into the three-volume
dioxin  series  was  done by PEDCo  Environmental,  Inc.,  Cincinnati,  Ohio,  with
Ms. M.  Pat Esposito  as Project Manager.    Technical  assistance was provided
by Ms. Diane N. Albrinck.
                                      xi

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                                  SECTION 1

                                INTRODUCTION
     A dioxin  is any of a  family  of compounds known chemically as  dibenzo-
para-dioxins.  Each of  these  compounds  has  as  a nucleus  a triple-ring  struc-
ture consisting  of  two  benzene rings interconnected to  each  other through a
pair of oxygen atoms.   Shown  below are the  structural formula  of the  dioxin
nucleus and  also the abbreviated  structural convention  used throughout  the
report series.
     Most environmental  interest  in  dioxins and most studies of  this family
of  compounds  have  centered on  chlorinated dioxins, in  which  the  chlorine
atom occupies  one or more  of  the eight substitution positions (Blair 1973;
Lee et al.  1973; Nicholson and Moore 1979).

     The interest  of health and  environmental researchers  in  chlorodioxins
arose principally  because of  the  toxicity  and distribution of one  of these
compounds,  2,3,7,8 tetrachlorodibenzo-p-dioxin  (2,3,7,8-TCDD),* whose struc-
tural formula is as follows:
   Throughout  this  report,  the  2,3,7,8-tetrachloro  isomer is  specifically
  noted as 2,3,7,8-TCDD to differentiate it from the other tetrachloro
  isomers.  In many  cases,  however, general reference  is  made  to the family
  of tetra  isomers  as  TCDD's  because of the difficulty in isolating specific
  isomers.  Refer to preface for further explanation.

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 This is an  unusual  organic chemical, symmetrical across both  horizontal and
 vertical axes.   It  is  remarkable for its lack  of  reactive  functional groups
 and  its  chemical stability.   It is a lipophylic molecule, virtually insol-
 uble in  water and  only  sparingly  soluble  in most  organic  liquids;  it is a
 colorless crystalline solid at room temperature.

      Although 2,3,7,8-TCDD was  first reported  in the chemical literature in
 1872, no major  investigations  into its  toxicity were  begun  until the 1950's.
 Because of the  remarkable  stability of this substance in biological systems
 and its extreme  toxicity,  cumulative effects of extremely  small doses are a
 major concern.   For example, the  ID™*  of  2,3,7,8-TCDD  for the male guinea
 pig has been  shown  to  be only 0.6  ug/kg  or 0.6 part  per billion body weight
 (McConnell  et  al.,  1978).    Fetal  mortality has been observed in rats that
 had  been  fed  10 consecutive  doses of  2,3,7,8-TCDD  at  the  level  of 0.125
 (jg/kg per day (World Health  Organization 1977).   It is  reasonable to pre-
 sume, therefore, that the  slightest trace of 2,3,7,8-TCDD in the environment
 may have adverse effects on  the  health  of both  human  and animal populations.

      In  view of  these considerations,  it is vitally  important to scrutinize
 carefully  the  probable  avenues  of contamination  of the  environment  with
 2,3,7,8-TCDD.   It has been  recognized for some  time that 2,3,7,8-TCDD can be
 produced  in  the manufacture  of  2,4,5-trichlorophenol.   Other dioxins  are
 similarly  produced  in  the  manufacture  of other chlorophenols.   The amounts
 of  dioxins  produced  depend  on  process  controls  such  as temperature  and
 pressure.   Since  dioxins may  be  present in these  and  other manufactured
 chemical  products,  it is also  likely that they  may be present in the chemi-
 cal  wastes  and sludges remaining from these  processes.  If  this is the case,
 indiscriminate  discharge  of these wastes  into the environment, or the use of
 improper  disposal procedures could  lead  to  the  contamination  of water, air,
 or  foodstuffs.    This might,  in  turn,  result in widespread exposure  of the
 population to  TCDD's  and  other  dioxins.

      Since  1972  the  personnel   of  the   Brehm  Laboratory  of Wright  State
 University  have  been performing  sensitive   dioxin  analyses  under  programs
 supported by  several  government agencies  (U.S.  Air Force,  U.S. Environmental
 Protection Agency (EPA),  U.S. Department  of  Agriculture),  and the states of
 Michigan, New  York,  and Arkansas.   In these investigations  Brehm  Laboratory
 has developed  and applied complex analytical methodology  for  the  determina-
 tion  of  TCDD's  in  many  types  of samples,  including  herbicides,  industrial
 chemicals, soils, water,  air, biological  tissues and  fluids (both human and
 other animal),  and combustion  products  and  related samples  (Taylor  et al.
 1973; Taylor,  Hughes,  and Tiernan 1974a,b,c; Fee  et al.  1975; Hughes  et al.
 1975; Taylor,  Tiernan, and  Hughes  1974;  Tiernan 1975a,b;  Tiernan,  Taylor,
and Hughes 1975;  Taylor  et al. 1975,  1976,  1977, 1979; Tiernan et al.  1979;
 Erk,  Taylor,  and Tiernan  1979;   Yelton,  Taylor, and Tiernan  1977;  Wright
State University  1976).   The  levels of  TCDD's   in these samples have  ranged
from  high parts per million  (ppm)  to low parts  per trillion  (ppt).   A  sig-
nificant number of samples examined  have been found to contain
  LD
  ce
3™:   The  administered dose  of  a substance  which is  lethal  to 50 per-
irtr of a test group  of  animals.

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detectable amounts  of  TCDD's.   On the basis of  these  findings many investi-
gators  believe that  TCDD's may  already be widespread  contaminants in  the
environment.

     The analytical techniques  applied  by Brehm Laboratory in  these  earlier
dioxin programs have  varied widely in terms of  the  complexity of equipment,
sample  preparation,  and  the  overall  sensitivity  and   specificity  of  the
procedures.   It  is now  apparent  that a single basic  technique,  amenable  to
minor  modifications,  would  be  desirable for  the  purpose of  characterizing
various  types of  chemical  samples,  provided  that  such  a  technique  could
satisfy  all  the specified  criteria  for sensitivity,  specificity,  and  other
analytical factors.

     Sensitivity in the  ppt range is required  because  of the  potent toxicity
of  2,3,7,8-TCDD.   The current  detection capability  is approaching 1 ppt  in
at  least  some sample  matrices  and must  be  developed in  others, particularly
chemical  process  wastes and sludges.   Accuracy is  also important  in  these
determinations, owing  to current  and potential regulatory actions that  hinge
on the analytical  data.

     The Brehm  Laboratory,  in  a subcontractual effort  with Battelle Columbus
Laboratories,  supported through  a  prime contract between  Battelle  and  the
U.S. EPA,  has undertaken development of new analytical techniques for use in
quantitating  ppt  levels of  TCDD's  in various  chemical wastes.   The  goal  in
this work  was to  develop a  unified  analytical  approach  to the handling of a
variety of chemical waste sample types and matrices.

     The  U.S. EPA  supplied 17  test samples  representing  various types  of
chemical  wastes  or residues  generated  during  the manufacture of  chloro-
phenols  and   related   chemicals.   These  samples  were  expected to  contain
TCDD's  and were  used in  methods  development by  the  Brehm  Laboratory ana-
lysts.    Presented  herein  are  the final  results  of  this work.   This volume
includes  a background  discussion of various  analytical  approaches to  the
detection  of  TCDD's,  the newly developed and  validated  analytical  method, a
description  of the procedures  used  in  development  of  the  method,  and  the
analytical  data  obtained  in  applying  the  method  to  various  industrial
samples.   Appendix A  of  this  report  discusses general  principals of  gas
chromatography  and mass  spectrometry.   Appendix  B  discusses  other  methods
and  procedures found  in  current literature  which may be  used  to detect
TCDD's  in a  variety   of sample  matrices.   Appendix  C  is  a  compilation  of
references on analysis of TCDD's, categorized by sample type.

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                                   SECTION 2

                             ANALYTICAL BACKGROUND*


      Analytical methods  for  detecting TCDD's  in various  types  of samples
 involve extensive  sample  preparation  procedures  followed by highly complex
 instrumental  analysis.   This  section discusses  various  approaches  to the
 detection  and  quantitative measurement of  TCDD's, which had been used prior
 to the inception of the present study in  1978.


 SAMPLE PREPARATION

      Because  TCDD's  may be  found in  a  variety  of  matrices  many different
 sample extraction/ preparation methods have been  developed.   Although they
 differ in  complexity, most of  these  methods may be classified into two major
 categories:   first,  those  characterized  by a  highly basic extraction step,
 and second,  those involving only  neutral extraction.  The neutral extraction
 technique  was  developed  to preclude  the  possibility that  treatment  with a
 strong base  might generate compounds  that  could form chlorinated dioxins in
 the mass  spectrometer.   Following extraction,  the  sample preparation steps
 are similar  for both techniques,  differing only in the method of application
 and complexity.  Both  extraction  procedures are  described  in detail  below.

 Basic  Extraction Method

     Historically  basic  extraction  methods were first  developed for  the
 determination  of TCDD's  in environmental  samples (Crummett and Stehl  1973;
 Baughman  and  Meselson  1973a;  Baughman  and Meselson  1973b).  Such  sample
 preparation  techniques  begin  with  digestion  of  a sample  aliquot  using
 alcohol and a  strong base.   This  is  followed  by  a series of organic solvent
 extractions to  separate the TCDD's from the alkaline mixture.   Solvents such
 as  ethanol, hexane,  petroleum ether,  and methylene  chloride have been used,
 either  singly  or in combination.   The  solvent extracts are combined and then
 subjected to a  series of washings with distilled water and strong acid.  The
washed  extract  is  then  treated   to  remove all traces  of water  and  passed
 through  one  or  more  chromatographic columns   for  removal  of  some  co-
 extractants,  primarily  polar  compounds.   Instrumental  analysis  follows.


*  Supplementary information on  analytical  methods for  detecting  dioxins  in
  various  types of samples may be  found in the appendices.

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     An  example  of  a  typical  basic  extraction/preparation  technique  for
nonfat tissue consists  of heating 10 g of sample with  10 ml of ethanol  and
20 ml  of  40  percent  potassium hydroxide solution for 30  minutes.   After  the
solution cools, an additional  10  ml of ethanol  is  added  and the solution is
extracted  with  four  10-ml  portions  of hexane.   The  preparation  procedure
consists of washing  the combined  hexane extracts with concentrated  sulfuric
acid  until  the acid fraction becomes  only slightly colored.  The  acid wash
is followed  by a  10 ml  water  wash,  followed  by  evaporation  to dryness  at
room  temperature  with  a  stream of  dry  air.   The sample  is  then redissolved
in hexane  and  further  purified  by elution  chromatography  using  sorbents such
as alumina,  silica gel,  or Florisil,  either singly or in combination.   The
final eluate is concentrated prior to analysis.

Neutral Extraction Method

     The  neutral  extraction  and preparation technique was originally  devel-
oped  by  O'Keefe,  Meselson, and Baughman  (1978).   Albro  and Corbett  (1977)
describe  an  alternative  neutral  extraction  method.  A  typical neutral  ex-
traction technique for  analysis of TCDD's  consists of extracting  the  sample
with  10  ml of  hexane.   The hexane  solution is then chromatographed  with a
magnesia-Celite 545  column,  an  alumina column,  an alumina minicolumn,  and
finally a  Florisil minicolumn.   The Florisil column is  eluted with methylene
chloride,  and  the eluate  is  concentrated  in  preparation for  analysis.   It
has been  asserted  that  neutral  extraction  methods are particularly effective
for  fish  tissues  and  human  milk (O'Keefe,  Meselson,   and Baughman  1978;
Harless and Dupuy 1979).

Chemical Composition of Extracts

     The  sample  preparation  techniques  described  above  are  useful  for
destroying the  integrity of  the  sample matrix and yield a small  volume of
organically  miscible/soluble   residue.   The net  effect  of these  clean-up
procedures is  the enrichment  of  the TCDD's relative  to  the natural  compo-
nents  of  the   sample   matrix,  as  well  as  other  chlorinated  environmental
contaminants such  as PCB's  and DDE.*  The  latter compounds are  often present
in  the  sample  in  significantly  greater  concentrations   than  the  TCDD's
(larger  by a factor of 106) and,  therefore, may  not be completely removed
from  the  extract  at  this point.   In addition,  it is unlikely  that the fore-
going   procedures  result  in separation  of 2,3,7,8-TCDD  from  its  other 21
TCDD isomers which may  have been present in  the sample.**
 * DDE,  or 2,2-bis-(p-chlorophenyl)~l,l-dichloroethylene,  is  commonly found
   in  environmental  samples;  it  is a  degradation  product of  the pesticide
   DDT.
** Subsequent  to the  completion  of the work described  herein,  reports have
   appeared in the literature which describe methods for synthesis and isola-
   tion  of the 22  TCDD isomers  (Nestrick 1979; Dow 1980).   Using  such new
   analytical  procedures it  is  now possible  to isolate  and  quantitatively
   determine 2,3,7,8-TCDD  in  environmental  samples even  in the presence of
   the other 21 isomers.

-------
      Consequently,   detection  and  quantisation  of  TCDD's  in  general  and
 2,3,7,8-TCDD  in  particular in  this "enriched" but  still  rather  chemically
 complex  extract  can  only  be  accomplished by  using a  highly specific and
 sensitive  instrumental  method.    The  method  of  choice,  and that described
 below, is  coupled gas  chromatography-mass  spectrometry.


 GAS  CHROMATOGRAPHIC  AND MASS  SPECTROMETRIC METHODS  OF ANALYSIS*

      Because of its ready  availability and relative  ease of  application, gas
 chromatography has  been extensively used  for  the  detection  and  quantitation
 of  TCDD's  (Elvidge  1971;  Williams and  Blanchfield 1971;  Firestone  et al.
 1972; Williams and Blanchfield  1972;  Crummett and  Stehl 1973; Edmunds, Lee,
 and  Nickels  1973; Webber and Box  1973; Buser  1976; Bertoni et  al.  1978).  In
 many  instances, the authors cited above  have  found that the chromatographic
 methods  lack  the  required specificity  for  determining  TCDD's  in  complex
 samples.    Consequently these  researchers  and  others  have sought more sensi-
 tive  and specific methods of  detection.

      At  present the analytical  method which  is almost  exclusively used for
 the  detection  and quantitation  of TCDD's  is coupled gas chromatography-mass
 spectrometry or GC-MS  (Crummett and Stehl  1973; Tiernan et  al.  1975; Taylor
 et  al.   1975;   Buser  and Bosshardt 1976;  Harless  1976;  Buser  1977;  Gross
 1978).

      GC-MS  is  the only known method that  can provide very  high sensitivity
 as well  as  the required selectivity  for  TCDD's.    A particularly sensitive
 and  specific  GC-MS technique  which  has  been  used  entails  low-resolution
 selective  ion  monitoring.   In  the case of TCDD's,  fragment  ions at nominal
m/e 320 and m/e 322, as  shown below, are monitored.
        Cl
>
k


"a
t


70 eV
electrons




\l
>'
c M 35



%|.
>
i
C1«Y " *"
                                                                     ©
                                                                       + e
                                                                      320)
                                                                     ©
                                                                       + e
                                                          321.8936 (non1n«l
                                                                         322)
  A discussion  of  the  principles of  gas chromatography  and mass spectrom-
  etry is  presented in the Appendix.

-------
The intensities  of  these ions are recorded as  the  TCDD's elute from the  gas
chromatograph.   The  ratio  of the  intensities  of m/e  320 to  m/e  322 is  a
characteristic indicator  of  TCDD's.   Unfortunately  other compounds  which  may
also  be  present  in  the  sample  extract  can  also give rise to  mass  spectral
ions  at  the  same  nominal  masses  (m/e  320  and m/e  322) as  TCDD's.   Two
approaches can minimize this problem.

     The  first  approach  utilizes high  resolution   mass spectrometry  (M/AM
>9000) to  increase  the selectivity.   The ions appearing under low-resolution
MS  conditions  at nominal  mass  322  may  be produced  from TCDD's which have
Ci2H4Cl402 as  their  elemental composition and  thus  have an  "exact"  mass  of
321.8936.   Interfering  ions such   as  pentachlorinated  biphenyls  may also
appear at  nominal mass 322,  but  their elemental composition is  C12H3C15,  and
therefore  they  have  an  "exact"  mass  of  321.8677.    Thus,   using  high-
resolution MS  these  ions of slightly different mass  are distinguishable,  and
so  the  dioxin component  having  the  exact  mass of  321.8936  can  be  reliably
measured.   Conceivably,  ions  having  the  C^^Cl^  composition  can   be
produced  from   other  compounds,  but proper   selection  of  chromatographic
procedures  maximizes  the  possibility  of separating   such  compounds from
TCDD's.   The  achievement  of  detection limits  in the  low-ppt range at high MS
resolution  generally  requires  the  use of  data acquisition methods  which
entail signal  averaging  (Shadoff and Hummel  1978; Gross 1978;  Taylor et  al.
1976).

     A second approach  to  the  problem of  separating  TCDD's   from  closely
related  interferences  makes use  of  low-resolution mass  spectrometry  but
incorporates  a  more  selective   separation  step prior  to the  mass  spectro-
metric analysis.  Capillary  column   gas  chromatography  is useful   for this
purpose  (Buser 1977),  but liquid chromatography followed by capillary column
gas chromatography  has proved even  more fruitful (NestHck,  Lamparski,  and
Stehl  1979; Dow  1980).

     In  both  the  GC-high -resolution and  the  GC-low-resolution mass  spec-
trometric  methods,   internal  standards  are   frequently  used  for the  quan-
tification of TCDD's.   The analytical method developed  in  the  present study
utilizes  an internal standard, namely 37Cl4-2,3,7,8-TCDD.

-------
                                   SECTION 3

                              ANALYTICAL METHOD*
     The  analytical  procedure ultimately developed and  described  herein for
determination  of TCDD's in various industrial process waste samples utilizes
two  separate  GC-MS systems.   A gas chromatograph coupled to a low-resolution
quadrupole  mass  spectrometer  (GC-QMS) is used for preliminary identification
of TCDD's  in  the extracts of the waste samples.   A second apparatus coupling
a  gas  chromatograph  and a high-resolution  mass spectrometer  (GC-MS-30)  is
used  to  confirm  the   results  obtained  with  the  GC-QMS  technique.   The
analysis  method  entails  two  steps,  sample preparation  and  instrumental
analysis,  as  described  below.   It  should be emphasized that,  even  with the
elaborate  separation techniques  employed here,  the  2,3,7,8-TCDD  isomer  is
still  not  resolved from the  other  TCDD  isomers  if these are present  in the
sample  extracts.   As  a result,  the  quantitative  data  obtained  here  for
TCDD's must be considered an  upper limit rather than an absolute level  for
any individual TCDD isomer.


SAMPLE PREPARATION

     The following  procedures were  developed as an approach to  preparation
of  industrial  waste  samples  and  have  been successfully  applied in  this
study.

     1.    Place a 2.0 g aliquot of the sample in  each of the two  extraction
          vessels.   To  each  aliquot,  add an appropriate quantity of  37C14-
          2,3,7,8-TCDD  dissolved   in   "distilled-in-glass"   benzene  as  an
          internal  standard.    Spike one  of  the  two  aliquots with  an addi-
          tional  known  quantity of  authentic native  2,3,7,8-TCDD at  a  con-
          centration  equal  to  the  nominal  amount expected  in the  sample.

     2.    Add  30 ml "distilled-in-glass"  petroleum  ether to each  sample  and
          mix  thoroughly.

     3.    Extract each  organic  solution with 50  ml of  double-distilled water
          and  discard the aqueous layer.

     4.    Extract each  solution with  50 ml of 20 percent potassium hydroxide
          and  discard  the aqueous basic layer.
  This section  presents the analytical  method only; discussion of  develop-
  ment of  the  method  follows  in Section 4.
                                    8

-------
     5.   Extract  each solution  with 50  ml  of  double-distilled water  and
          discard the aqueous portion.

     G.   Extract each  solution  with 50  ml  of concentrated sulfuric  aci,d  and
          discard the aqueous acidic layer.

     7.   Repeat step 6 until the acid layer is nearly colorless.

     8.   Extract each  organic  solution  with 50 ml  of double-distilled water
          and discard the aqueous layer.

     9.   Dry each organic solution over anhydrous sodium sulfate.

    10.   Quantitatively  transfer  each organic  solution  to another  vessel,
          and  concentrate to  a volume of  approximately 1  ml  by passing  a
          stream of  purified nitrogen over the surface of  the  liquid while
          applying gentle heat (50°C) to the vessel.

    11.   Construct  a  chromatography  column  for  each  sample  by packing  a
          disposable  glass pipette  (I.D.=  0.8 cm) with glass wool  and 2.8 g
          of Woelm basic  alumina (previously activated by maintaining it at
          600°C  for  a minimum of 24 hours, then cooled in  a dessicator  for
          0.5 hour prior to use).

    12.   Quantitatively  transfer  each concentrated  organic solution  to  the
          top of a column.

    13.   Elute  each column  with  10 ml  of 3 percent  "distilled-in-glass"
          methylene  chloride  in  "distilled-in-glass"  hexane, and discard  the
          entire column effluent.

    14.   Elute  each column with 20 ml  of 20 percent methyl ene  chloride in
          hexane and collect the eluate in  four 5-ml  fractions.

    15.   Elute  each column with 10 ml  of 50 percent methylene  chloride in
          hexane and retain the entire column eluate  for analysis.

    16.   Elute  each column with  3 ml of  50 percent methylene  chloride in
          hexane and retain the eluate for  analysis.

    17.   Concentrate all  six  fractions  in benzene to an  appropriate  volume
          (usually 0.1 to 1.0 ml) and proceed with analysis.
INSTRUMENTAL ANALYSIS

     The application  of  GC-MS instrumentation methods for analysis of TCDD's
requires knowledgeable  and experienced  personnel,  dedication  of  the equip-
ment,  and   significant   capital  and  operating  costs.   The requirement  for
detecting  low ppt  levels of  TCDD's  in these  analyses necessitates  such  a
sensitive  and selective  analytical  method.  Because  this is  currently  the

-------
 only known method  which  meets these criteria,  the  relatively  high expense is
 unavoidable.

      The  following is a  brief description of  the instrumentation required
 for the analytical  procedures developed herein.

 GC-QMS System

      The  GC-QMS  system  consists  of  a Varian Model  2740  Gas Chromatograph
 coupled  directly  (no  helium  separator  is   required)  to  an Extra-nuclear
 Quadrupole Mass  Spectrometer.  The  GC was  adapted  to  include  a sophisticated
 system  of remotely  actuated high-temperature switching valves  (Valco Co.)
 and Granvilie-Phi Hips  molecular  leak  valves,  so  that  the  column effluent
 could be  readily regulated  (Tiernan  et al. 1975a;  Erk,  Taylor,  and Tiernan
 1978).

      With this arrangement,  the total  column  effluent  can be directed into
 the mass  spectrometer  ion source,  or the effluent  flow can be  split,  one
 portion  going  to  the  ion  source   and  the other  to a  gas  chromatographic
 detector, as desired.   The  use of  a differential  high-speed pumping system
 on  the  source  vacuum envelope permits  introduction of as  much  as  65 ml/min
 of  effluent  from  the  gas  Chromatograph  into  the  mass   spectrometer  ion
 source.    Admitting the  total Chromatograph  effluent into  the mass  spec-
 trometer source enhances  the  sensitivity of the analysis.

      For purposes  of instrument  control   and  data acquisition,  the  GC-QMS
 system  is coupled to an  Autolab System IV Computing  Integrator.   Additional
 capacity for  off-line  data  reduction  is  available  with  a  Hewlett-Packard
 2116C  Minicomputer, which is  programmed to accept  data  (punched paper tape)
 from the system when  necessary.

 GC-MS-30 System

     The GC-MS-30 system used  in these studies consists of a Varian 3740 Gas
 Chromatograph  coupled through  an  AEI silicone membrane  separator  to  an  AEI
 MS-30 Double-Focusing,  Double-Beam  Mass Spectrometer.   The mass spectrometer
 is  equipped with a unique electrostatic analyzer  scan circuit  developed by
 Wright  State University,  which permits the  monitoring  of  as many as  four
 mass peaks,  essentially  simultaneously, by rapidly and sequentially stepping
 and  switching  between  the  masses   of interest,  while maintaining  picogram
 sensitivity  for  TCDD's.   The  data are recorded by  use  of  a Nicolet  1074
 Signal Averaging  Computer.

Sample Analysis

     Analysis consists of three steps  as described below.

     1.    Analyze  each eluate fraction  (collected  in  the elution  chromatog-
raphy  separation  of  the  sample)  on  the   low-resolution  GC-QMS,   using  the
following operating parameters:
                                    10

-------
     Van"an 2740 Gas Chromatograph
          Column:
          Carrier gas:
2 m  x 3  mm  I. D.  glass packed with  3  percent
OV-7 on Gas Chrom Q

Helium at 65 ml/min (the total  chromatographic
column effluent is  admitted  to  the mass spec-
trometer ion source)
          Temperatures:   Injector:   255°C
                         Column:     275°C
                         Transfer line:   295°C
     Quadrupole mass spectrometer

          Ionizing voltage:

          Multiplier:

          Resolution:

          Source envelope pressure:

          Analyzer envelope pressure:

          Masses monitored:

          Source temperature:

          Analyzer temperature:
               23.5 eV

               3200 V

               1:350

               1.4 x 10"4 torr

               8.0 x 10"6 torr

               m/e 320, 322

               250°C

               120°C
     2.   Confirm  any samples showing positive  levels of TCDD's on the  low-
resolution  GC-QMS  by analysis  of the  corresponding  eluate fractions  using
high-resolution GC-MS-30 and the following operating parameters:

     Varian 3740 gas chromatograph

          Column:         1.8  x  2  mm I.D.  coiled glass  column  packed
                         with  3  percent  Dexsil  300  on  Supelcoport
                         (100/120 mesh)

          Carrier gas:    Helium at a flow rate of 30 ml/min

          Temperatures:   Injector:  250°C
                         Column:    240°C
                         Transfer line:   285°C
                                    11

-------
     AEI MS-30 mass spectrometer

          Resolution           1:12,500

          Ionizing voltage:    70 eV

          Masses monitored:    m/e  319.8966,  321.8936,  325.8805,  and
                               327.8846

          Temperatures:        Membrane separator:  215°C
                               Transfer line:  270°C
                               Source:  250°C

     3.   Determine  the  overall   recovery  of  the  analytical  procedure  by
measuring the amount  of  internal  standard  (37Cl4-2,3,7,8-TCDD)  recovered.
                                    12

-------
                                  SECTION 4

                           DISCUSSION AND RESULTS
     For use  in  developing  and demonstrating the analytical methodology  for
determination of  ppt levels  of  TCDD's in  process  wastes and related mate-
rials, samples were  provided  that were representative  of wastes  from  several
different  industrial  chemical processes  that  might be  expected to generate
chlorodioxins.  The  samples were  obtained by the U.S.  EPA  from  plants manu-
facturing  trichlorophenol,  pentachlorophenol,  and hexachlorophene, and  from
plants processing wood  preservatives.   Initially, the  nature and identity of
each sample were  unknown  to the Wright State  investigators, although infor-
mation was made  available  early  in  the  program  about  two  of  the  samples
originating  from  trichlorophenol  manufacturing  processes.   Subsequently,
identifying  data on  most  of the  remaining  samples  were  obtained  and  are
summarized in Table 1.

     Because  still  bottom  samples collected  at a trichlorophenol manufac-
turing plant  were considered of major interest, a  sample of this type  (EPA
sample 2) was selected for use in preliminary investigations.

     The  initial  approach  to analytical  method development,  based  on  the
experience of  Wright State personnel  in chlorodioxin  analysis,  is outlined
below.

     1.    If the  sample  is  solid,  dissolve a portion in an  immiscible combi-
          nation  of  aqueous  and  organic  solvents,  such as water  and petro-
          leum ether.   If the sample  is  a liquid,  extract a portion of  the
          material  with  a  similar  water-organic  solvent system.   In  the
          absence of  any prior  knowledge about the  content  of TCDD's  in a
          given sample, the quantity  to be extracted must be  selected on the
          basis  of  sensitivity  of the  overall technique  (as  indicated by
          previous experience) and the desired limits of detection.

     2.    Separate the  aqueous  component of the sample-solvent  mixture  from
          the organic phase  and discard the aqueous portion.

     3.    Extract  the  organic  fraction  with  sequential  washes  of  acid,
          water,   base,  water, acid,  and  water (in that  order),  and  discard
          the washes.

     4.    Concentrate the remaining organic phase  to  near dryness  and elute
          through an  alumina  column,  using appropriate  solvents  to  separate
          the TCDD's and other sample components.
                                    13

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TABLE 1.   SAMPLES USED IN DEVELOPMENT OF ANALYTICAL
      METHOD FOR TCDD'S IN INDUSTRIAL WASTES
EPA No.
C04130
C04131
C04132
2
3
4
5
6
12700
12701
12702
11020
11021
11022
11023
11024
11025
Sample type
Liquid slurry
Solid
Liquid
Liquid/sol id
Slurry
Slurry
Liquid/solid
Liquid
Liquid/solid
Liquid
Solid
Liquid/solid
Liquid
Liquid/solid
Solid
Solid
Solid
Source and identity of sample
Givaudan: aqueous slurry of hexachlorophene
Givaudan: activated clay filter cake from
hexachlorophene manufacturing
Givaudan: ethyl ene di chloride recovery solutio
from hexachlorophene manufacturing
Transvaal: still bottom from trichlorophenol
(TCP) manufacturing
Transvaal: cooling tank bottom from TCP manu-
facturing
Transvaal: discharge line from TCP manufactur-
ing
Transvaal: sludge from TCP manufacturing
Transvaal: type unknown; presumably TCP proces
sample
Reichold Chemical: sludge from intake of settl
ing pond, pentachlorophenol (PCP) manufacturing
Reichold Chemical: sludge from discharge of
settling pond, PCP manufacturing
Reichold Chemical: PCP manufacturing
Baxter: retort solids residue from wood pre-
serving
Baxter: storage tank solution from wood pre-
serving
Baxter: cooling water solids from wood pre-
serving
Baxter: treated wood from wood preserving
Baxter: soil from neighborhood of wood preserv-
ing plant
Baxter: sludge from wood preserving
                       14

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     5.   Concentrate  the  fraction  containing  TCDD's   and  subject  it  to
          preliminary  screening  analysis  by  use  of  the  GC-QMS  system,
          operated  in  the  selected-ion  monitoring mode  and  adjusted  to
          detect  m/e  322 and  m/e 320,  the  two most abundant  peaks in  the
          isotopic molecular ion cluster of 2,3,7,8-TCDD.

     6.   If  the  initial  screening  indicates  a  positive  level of  TCDD's,
          then  the level  must be  confirmed  and quantitated by use of  the
          GC-MS-30 system.

     This approach was used  in analysis of sample  2.   Subsequent  modifica-
tions  of this  initial procedure and  other  observations  are   discussed  in
following subsections.


DEVELOPING SAMPLE PREPARATION TECHNIQUE

     Four aliquots  of sample  2 were  extracted with a mixture  of  water  and
petroleum ether.   The aqueous  portion was discarded, and each  organic  frac-
tion was  washed successively  with acid, water,  base, water, acid,  and water.
The  samples  were then  concentrated and  transferred to  a  2.8 g Woelm  basic
alumina column  (length 12 cm,  internal diameter 0.8 cm).

     Large  quantities  of a  white  crystalline  substance   appeared  in  the
column  eluate.    The  column  apparently was  overloaded   owing  to   the  large
quantity  of  this material present in  the  sample.   This substance possibly
accounted for  interference in  the mass chromatogram (Figure 1).  Adjustments
of  the column  chromatography  procedure were  therefore  made in  an  effort to
eliminate  this  crystalline  contaminant  in  the  fraction  containing  the
TCDD's.

     A  solvent  screening study  was done  to  evaluate the  solubility of  the
contaminant  and  the   potential   for  its  removal  from   the  sample  matrix.
Results are as  follows:

          Solvent tested                Solubility of contaminant

          100%  methanol            Slight solubility

          3% methylene             Solubility slightly greater than
           chloride in hexane       in 100% methanol

          25% carbon tetra-        Solubility slightly greater than
           chloride in hexane        in 3% methylene chloride in  hexane

          100%  methylene           Completely soluble
           chloride

     Next, elution characteristics  of  the  alumina  column were  evaluated.
Table  2  presents the  solvents  and  the  discrete  fractions   collected  in
determining the elution  characteristics  of the Woelm basic  alumina column.
                                    15

-------
                      TCDD'S
TIME-
      Figure 1.   Mass chromatogram of extract of
      sample 2,  at m/e 322 obtained with GC-QMS.
                           16

-------
             TABLE 2.   ELUTION OF TCDD'S IN EXTRACTS OF SAMPLE 2
Set No.
    Eluting solvent
Total volume
 of column
effluent, ml
     Volume of
    fraction(s)
     collected
  Al


  A2
  Bl


  B2
  Cl


  C2
  Dl


  02
3% methylene chloride in
 hexane

50% methylene chloride
 in hexane
3% methylene chloride in
 hexane

20% methylene chloride
 in hexane
     10
     13
     10


     18
25% carbon tetrachloride
 in hexane

50% methylene chloride
 in hexane
25% carbon tetrachloride
 in hexane

20% methylene chloride
 in hexane
     10


     13
     10


     18
total  10 ml
1st 5 ml in one
 sample; 6th through
 13th ml in separate
 1-ml fractions

total 10 ml
1st 5 ml in one
 fraction 6th through
 13th ml in separate
 1-ml fractions; 14th
 through 18th ml in
 one fraction

total 10 ml
1st 5 ml in one
 fraction; 6th through
 13th ml in separate
 1-ml fractions

total 10 ml
1st 5 ml in one
 fraction; 6th through
 13th ml in separate
 1-ml fractions; 14
 through 18th ml in
 one fraction
                                      17

-------
      Selection of the  solvents  and the eluate fractions was  based  on earlier
 experience  of  Brehm  Laboratory  personnel   in  column  chromatography with
 similar sample matrices.

      The  eluate  fractions  were analyzed  for TCDD's  by use  of  the  GC-QMS
 system.    The results,  presented  in  Table  3,  show  clearly that  the best
 elution sequence  involves  the use of  10 ml  of 3 percent methylene chloride
 in hexane,  followed  by 18  nfl of  20  percent methylene  chloride  in hexane.
 This  sequence yields TCDD's  in  a well-defined fraction containing  few other
 contaminants.   Use  of all  the  other solvent pairs  yielded fractions that
 generated  interferences in  the  dioxin mass chromatogram which were as great
 as those shown in Figure  1 or greater.

 Application  of Initial  Procedure to EPA Samples

      The extraction and sample  preparation procedure  developed for sample 2
 was applied  to  ten  of  the other  industrial  samples supplied by EPA.   In
 these  analyses some interferences  were still  present  in the  extract fraction
 which  was  thought to  contain the  TCDD's;  the interferences  resulted in a
 higher minimum detection  limit  (ppb)  than was desired.   Portions of these
 samples  were  also spiked  with  known  quantities  of  2,3,7,8-TCDD  so that
 recoveries for the procedure could be determined.    The  recovery  in  GC-QMS
 analysis of  sample  2 was  127 percent.

     Surprisingly,  in  analysis  of  the other ten  samples by  the  same pro-
 cedure,  none of  the  added  2,3,7,8-TCDD was  recovered.   The same procedure
 was  then  applied  in  analyses of spiked aliquots of  these samples, but this
 time  all  the eluate fractions  from the alumina  columns were  retained  and
 analyzed for TCDD's.   Again,  no  2,3,7,8-TCDD was detected.   It was necessary
 to  further investigate  the sample preparation  procedures.

 Optimizing Sample Preparation  Procedure

     Another  sample (C04131)  was subjected to the  general  preparation pro-
 cedure  already described, up to the  point of elution  of the column.   Then
 the sample was spiked  with  a large quantity  of  2,3,7,8-TCDD by introducing
 it  directly  onto  the  alumina  column.   The  column  elution  characteristics
 were  then  evaluated as before  and  the results are shown  in Table 4.   This
 procedure  was repeated for  all  other   samples and their  column elution pro-
 files were determined.

     This  study  indicated  that  a  general  extraction and preparation pro-
 cedure must  include a provision for assessing the elution characteristics of
 the alumina  column  for  each type of sample matrix.   Apparently, each type of
 sample  conditions or  deactivates  the  column  in a  manner  peculiar to  its
matrix,  and  this  conditioning in turn, determines  the elution characteris-
tics of TCDD's, which may differ markedly in different sample types.
                                    18

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                 TABLE 3.  CONTENT OF TCDD'S IN COLUMN FRACTION FOR SAMPLE 2a
Solvent
set No.

AT
A2
Bl
B2
Cl
C2
Dl
D2
Eluate fraction no.
1
2
3
4
5
6
7
8
9
10
n
12
13
14
15
16
17
18
TCDD's detected
_*
+*
-
+*
0
+*
0
+*
_*
+*
-
+*
0
+*
0
+*
_*
+*
-
+*
0
+*
0
+*
_*
+*
-
+*
0
+*
0
+*
_*
+*
-
+*
0
+*
0
+*
_*
+*
-
+
0
+*
0
+*
_*
+*
-
+
0
+*
0
+*
_*
+*
-
+
0
+*
0
+
_*
+
-
+
0
+*
0
+
_*
-
-
+
0
+*
0
+
0
-
0
+

+*

+
0
-
0
+

+*

+
0
-
0
+

+*

+
0
0
0
+

0

+
0
0
0
+

0

+
0
0
0
+

0

+
0
0
0
+

0

+
0
0
0
+

0

+
.  Aliquots of EPA sample 2.
  Fraction numbers refer to those collected from each of the columns, as indicated in Table 2.
+ =

0 =
* =
TCDD's present in fraction.
No TCDD's detected in fraction.
Fraction not analyzed.
Two or more peaks evident in mass chromatogram near 2,3,7,8-TCDD retention time.

-------
TABLE 4.   RECOVERY OF 2,3,7,8-TCDD SPIKE FROM ELUATES OF SAMPLE C04131
Solvent
10 ml 3%
methyl ene
chloride in
hexane
20 ml 20%
methyl ene
chloride in
hexane
10 ml 50%
methyl ene
chloride in
hexane
No. of
fractions
collected
1


4

1

Volume
of each
fraction
10 ml


5 ml

10 ml

Action
Discarded


Analyzed by
GC-QMS

Analyzed by
GC-QMS

Results



No 2,3,7,8-
TCDD

80% 2,3,7,8-
TCDD
recovered

                                  20

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ANALYTICAL PROCEDURE

     Research  workers  in several  laboratories,  including the Brehm  Labora-
tory, have analyzed  various  types of samples for dioxin content.   Generally,
the  analytical approach  to  determining  a  chlorinated hydrocarbon of  this
type  in a complex sample matrix  has involved  quantitation  of  the  chloro-
carbon  by  use  of  electron capture-gas chromatography (EC-GC) or  gas  chroma-
tography-mass   spectrometry   (GC-MS).    The  studies   at  Brehm   Laboratory
entailed  use  of  GC-MS  and  high-performance  liquid chromatography  (HPLC).

GC-MS System

     As  described in  Section  3,  the  GC-QMS  system  was used  for  initial
detection of TCDD's  in the fractionated sample.   Then  the GC-MS-30 was  used
to confirm the positive levels of TCDD's detected in the GC-QMS.

     In  one  procedural  modification, a  labelled internal  standard,  37C14-
2,3,7,8-TCDD,  was added  to  all  samples.   Also,  the  MS-30  high-resolution
mass  spectrometer was  modified  to  permit essentially simultaneous step-
scanning  of  four ions  in  the  high-resolution  mode.    The  ions  typically
monitored were:

     m/e 319.8966,  a  major  molecular ion  in  the mass  spectrum  of 2,3,7,8-
                    TCDD

     m/e 321.8936,  a  major  molecular ion  in  the mass  spectrum  of 2,3,7,8-
                    TCDD

     m/e 325.8805,  a molecular ion indicative of interfering PCB's

     m/e 327.8846,  a  major  molecular  ion  in  the  mass spectrum  of  37C14-
                    2,3,7,8-TCDD.

High-Performance Liquid Chromatography (HPLC)

     In  earlier   studies  aimed   at  determining  TCDD's  in environmental
samples, concern  has  been raised that the  presence of  the  so-called  pre-
dioxins  (for  example,  polychlorinated  phenoxyphenols)  in the samples would
lead to false  positive  determinations  of TCDD's  because the latter  can  be
formed  by cyclization  reactions of the predioxins  in the hot injection  port
of gas  chromatographs.   The  present investigation ruled  out  potential false
positive effects  of  predioxins  by applying an  HPLC analytical technique  as a
quality  assurance measure.   HPLC does  not entail  injection of  the  sample
into  a  heated port   and  therefore  minimizes  the possibility   of  thermal
cyclization of predioxins.

     The HPLC  instrument  used  in these studies  is  the  Model  LC  5021  Varian.
This microprocessor-controlled HPLC  is both  completely automatic and  pro-
grammable and  incorporates a multiple  solvent system.    Three detectors are
available:   a  fixed-wavelength  UV  (254 nm) detector,  a variable-wavelength
UV detector,  and  a fluorescence detector.   A cathode ray tube (CRT) keyboard
                                    21

-------
 unit  displays  operating  parameters  while  a micropressor -based computing
 integrator (DCS-111L)  stores  the  data  and  performs appropriate calculations.
 The parameters applicable to  the  instrument  as it was used  in this study are
 listed below:

      Column:              DuPont Zorbax  ODS  (25 cm x 6.2  mm)

      Temperature:         50°C

      Starting  Pressure:   952 psig

      Solvent:             100%  Methanol

      Flow rate:           2.5 ml/min

      Detector:            UV (235 nm)

      Sensitivity:         0.02  absorbance  units full scale/15 ng TCDD's

 Upon injection  of a 10  ul  aliquot of the  sample 2 extract  into  the HPLC, a
 chromatographic  peak  having   a  retention  time  which was  the same  as  that
 observed  with  the  2,3,7,8-TCDD standard was observed.   Representative  HPLC
 chromatograms  are shown  graphically  in  Figures  2  and  3,  and these results
 indicate  a readily  detectable level  of TCDD's in the sample  2  extract.   It
 is  apparent that  the TCDD's detected cannot  have  been  formed by cyclization
 of  predioxins.

 Analytical  Results

      Attempts  were  made to extract  15  of the 17 EPA  samples  by  the  pro-
 cedures described  in section   3.  The remaining two samples, 11023 and .12702,
 were  not  subjected  to  these   methods.   Sample 11023 was a  section  of wood,
 which  the earlier experience   of Wright State had  shown  is not amenable to a
 potassium   hydroxide  digestion  process.   Sample  12702  was  not  analyzed
 because of  insufficient  time during the contract period.

     Twelve of  the  fifteen samples were  successfully  analyzed by the Wright
 State  procedure,  with  results  as shown in Table 5.   These data show that the
 procedure  is applicable to samples exhibiting a wide range of concentrations
 of  TCDD's  from  ppt to ppm (a  factor  of 106).  For those samples  in which no
 TCDD's were detected,  the minimum detectable  concentration  of TCDD's was in
 the  low ppt range (45 to 140 ppt).

     Examples of mass  fragmentograms  obtained with  the GC-MS-30  high resolu-
 tion mass  spectrometer  are shown in the following figures.   Figure 4 shows a
 four-ion   step-scan  mass  fragmentogram  of  benzene,  the  solvent used  for
dilution  of the  final  sample  residue.    Analysis  of  a  solvent blank  is
 repeated  before  analysis of each  sample in  order  to ensure  that no TCDD's
are  carried over  in the  injection syringe.   Figure  5  illustrates  similar
data  obtained  from  injection   of  a  sample  consisting  of  50  pg of native
2,3,7,8-TCDD and  1  ng  of  37Cl4-2,3,7,8-TCDD.  Note that  different  attenua-
                                    22

-------
                     [1
                            TCDD'S
                               I
                       I	I     1
     TIME
Figure 2.   High pressure liquid chromatogram of sample 2,
                            23

-------
                       2,3,7,8-TCDD

                            I
 TIME
Figure 3.   High pressure liquid chromatogram of
            2,3,7,8-TCDD standard.
                       24

-------
      TABLE 5.  RESULTS OF GC-MS-30 ANALYSIS OF EPA SAMPLES FOR TCDD'S
EPA sample no.
C04130
C04131
C04132
2
3
4
5
6
12700
12701
12702
11020
11025
11021
11022
11023
11024
Origin
Givaudan
Givaudan
Givaudan
Transvaal
Transvaal
Transvaal
Transvaal
Transvaal
Reichold
Reichold
Reichold
Baxter
Baxter
Baxter
Baxter
Baxter
Baxter
Quantity of
TCDD's found
ng/g (ppb)
NDa
ND
ND
40,000
675
22
070
ND
ND
ND
b
ND
ND
c
c
b
d
Minimum detectable
concentration
pg/g (ppt)
140
70
50
e
e
e
e
50
80
75

140
45




.  ND:   no TCDD's detected in excess of the minimum detectable concentration.
  Not processed.
^ General procedure could not be successfully applied to these samples.
a Not analyzed on GC-MS-30.
e An exact minimum detectable concentration was not recorded for these
  analyses; however the reported values for quantity of TCDD's found are
  well above the criterion of 2.5X noise.
                                     25

-------
ATTENUATION: 512
      m/e  319.8966
      Figure  4.   Four-ion  mass  fragmentogram of
    benzene solvent  blank  obtained with GC-MS-30.
                          26

-------
   ATTENUATION: 256
            m/e 321.8936

                 I
    m/e 319.8966
m/e 327.8846
                                   ATTENUATION: 8192
Figure  5.   Four-ion  mass fragmentogram of  50 pg 2,3,7,8-TCOD and
          1 ng 37Cl4-2,3,7,8-TCDD  obtained  with GC-MS-30.
                                 27

-------
 tions  have been applied to the various peaks displayed in Figure 5.  Figures
 6  and  7 demonstrate  similar  four -ion step -scan mass fragmentograms obtained
 for  two  of the EPA  samples.  Although  the fragmentogram for  sample  12700
 shows  peaks  at m/e  319.8966 and  m/e  321.8936,  their intensities  are  not
 greater  than 2.5  times  the  background;  this is one  of  the  criteria applied
for establishing the  presence  of TCDD's in a  sample.
of 37Cl4-2,3,7,8-TCDD  from sample  12700,  the minimum
tion (MDC) of TCDD's is 80 pg/g.
                                                        Based on the recovery
                                                        detectable concentra-
     The  mass  fragmentogram for sample 5  (Figure  7)  shows peaks at both m/e
 319.8966  and m/e  321.8936,  and the  intensities are well  in excess  of 2.5
 times  the background levels.  After  application of  a recovery correction on
 the  basis of the  internal  standard,  these data indicate  that  sample  5 con-
 tains  70  pg TCDD's  per gram  of  sample.   Data  similar  to those  shown  in
 Figures  4 through  7 were obtained  for the other  samples analyzed  in this
 program.

     Analyses  of  samples  11021  and  11022 were not  completed owing  to the
 formation  of  an  intractable  emulsion  at  the petroleum/ ether  interface.
 Analysis  of  sample 11024 on the GC-MS-30  system was  not attempted because a
 colored  residue was  visible in the  final  extract.   Earlier  experience had
 shown  that  such  residues  indicate that  the  sample  extract  contains gross
 quantities of compounds  other than TCDD's, which  lead  to  serious contamina-
 tion of the  high-resolution mass spectrometer.

     All  data in  Table 5 were derived from analyses  with the high resolution
 GC-MS-30  system.    For  each of  the  industrial  process samples,  the  appro-
 priate  elution   chromatogram fractions  to be  analyzed were determined  in
 advance  in  a  series  of alumina  column  elutions  using  an  aliquot  of the
 sample  spiked with  2,3,7,8-TCDD  standard;  these  elutions  were  accomplished
 in  a manner similar  to that described  for sample  2.   These  elution test
 samples were analyzed with  the low resolution  GC-QMS system.   Data pertinent
 to the  determination of  the elution characteristics  of TCDD's in the various
 samples are  shown  in  Table  6.   The  fractions  collected for each  sample  in
the elution experiments are as follows:
     1.


     2.


     3.


     4.


     5.


     6.
          Fraction
          chloride
I
in
- First
hexane.
5-ml portion eluted with  20 percent methylene
         Fraction  II  - Second 5-ml portion eluted with 20 percent methylene
         chloride  in  hexane.

         Fraction  III - Third 5-ml portion eluted with 20 percent methylene
         chloride  in  hexane.

         Fraction  IV  - Fourth 5-ml portion eluted with 20 percent methylene
         chloride  in  hexane.

         Fraction  V - First TO-ml portion  eluted  with 50 percent methylene
         chloride  in  hexane.

         Fraction  VI  -  Last  3-ml portion  eluted  with 50 percent methylene
         chloride  in  hexane.
                                    28

-------
                                                                                           m/e 327.8846
10
           ATTENUATION: 512
                         m/e 321.8936
              m/e 319.8966
                                                            ATTENUATION: 8192
                                                                      m/e 321.8966
                                                          m/e 319.8966
                                                                  1
              Figure 6.  Four-ion mass fragmentogram of sample 12700 obtained with GC-MS-30.

-------
co
o
       ATTENUATION:  512
m/e 319.8966
                          m/e 321.8936
                                                                                         m/e  327.8846

                                                                                               I
                                                                                               t
                                                             ATTENUATION:  4096
                 Figure 7.  Four-ion mass fragmentogram of sample 5 obtained with GC-MS-30.

-------
              TABLE 6.  TCDD ISOMER CONTENT OF COLUMN FRACTION
                      SAMPLES SPIKED WITH 2,3,7,8-TCDD
EPA
samples
C04130


3




12700


12701

11020


11024d


11025

Eluate .
fraction
IV
V
VI
V
III
IV
V
VI
IV
V
VI
IV
V
IV
V
VI
IV
V
VI
IV
V
Quantity
of 2,3,7,8-TCDD
added to
sample,
ng/g
10.42


10.35
50.64



12.14


12.84

9.86


3.71


6.54

Quantity
of 2,3,7,8-TCDD
detected in
fraction,
ng/g
ND
10.62
ND
597
ND
46
625
ND
ND
8.4
ND
ND
10.12
0.56
8.68
ND
0.29
1.09
ND
ND
5.63
Minimum
detectable
concentration,
ng/g
0.5

0.5

3.0


3.0
0.3

0.57
0.28
-


0.23


0.08
0.14

Recovery, %

102







69


79
6
88

8
29


86
.  See Table 4-1 for description of sample.
  Designation of eluate fractions:
    III  Third 5-ml aliquot eluted with 20% methylene chloride in hexane.
     IV  Fourth 5-ml aliquot eluted with 20% methylene chloride in hexane.
      V  First 10-ml aliquot eluted with 50% methylene chloride in hexane.
     VI  Last 3-ml aliquot eluted with 50% methylene chloride in hexane.
  ND:   no 2,3,7,8-TCDD detected in excess of the minimum detectable concentration.
  Portion of sample was lost during preparation.
                                     31

-------
      These fractions were  analyzed  with the GC-QMS in reverse  order, begin-
 ning with  the last  fraction  and continuing  backward  until the quantity of
 TCDD's detected  in the  several  fractions  was a reasonably large percentage
 of that originally  added as  the spike, or  until a  fraction was  reached that
 contained  no  TCDD's.   The data  in Table  6 show that TCDD's are completely
 eluted from all  samples  prior to Fraction  VI.   In most cases the bulk of the
 TCDD's appeared  in  Fraction  V,  although  in samples  11020  and  11024 the
 TCDD's were detected in Fraction IV.

      Table  7   summarizes the  total  recoveries of the  added  2,3,7,8-TCDD
 spikes achieved by  collecting  the  optimum  column chromatography  fractions of
 the various industrial  process  samples.   These  recoveries  range from 60 to
 102 percent, with a mean  value of 85 percent.

      Except for sample 2,  all  of the  samples  processed in this  investigation
 were also  spiked  with   37Cl4-2,3,7,8-TCDD.   This  compound  was  added  as  an
 internal   standard   in  the  analyses  with   the  GC-MS-30  system.   The  mean
 recovery   of   37Cl4-2,3,7,8-TCDD   for   the   samples  analyzed  herein was  74
 percent with  a  standard  deviation  of 16.8 percent.  The  recovery  data are
 shown  in Table 8.

 Confirmation of TCDD's  in Sample  2

     Measurements  in which m/e 320 and m/e  322 were monitored by  the low-
 resolution  GC-QMS system indicated that sample  2  contained  approximately 40
 ug  TCDD's  per gram  of  sample.   The  report  of this  high level of TCDD's
 prompted  considerable  concern both  at EPA and state  regulatory  organiza-
 tions.

     This  finding was  also controversial   because an  earlier  examination of
 this  sample in an  EPA  laboratory had yielded no indication of the  presence
 of  TCDD's.   It  was obviously  important,   therefore,  to more  definitively
 confirm the initial  Wright State analyses  of sample 2; this was done  by a
 procedure  essentially  the same  as that  which  is  described  as the  final
 method  (Section 3).

     The  sample  was extracted,  and  the  extract   was  subjected to  liquid
 chromatography  preparation.   As mentioned earlier,   the fraction  of  sample 2
 that was  eluted  from the alumina column with 20 percent methylene  chloride
 in  hexane  was   determined to  contain  the  bulk  of  the TCDD's.   Accordingly,
 this fraction  was analyzed for TCDD's by the GC-MS-30 system operated in the
 dual-ion  monitoring mode (m/e 319.8966 and 321.8936 were  monitored).   The
 resolution  of  the MS-30 mass spectrometer  was adjusted  to  1:12,500  for this
measurement.

     The  dual-ion  step-scan  mass  fragmentogram obtained with  this  sample
extract  is  shown  in  Figure  8  and  corresponding  data obtained  with  an
authentic  2,3,7,8-TCDD  standard  are  shown   in Figure  9.   For  EPA sample  2,
                                    32

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           TABLE 7.  RECOVERIES OF 2,3,7,8-TCDD-SPIKED SAMPLES
                 FOLLOWING ALUMINA COLUMN CHROMATOGRAPHY
EPA
sample no.
C04130
4
5
6
12700
12701
11020
11024
11025
Quantity of 2,3,7,8-TCDD Quantity of 2,3,7,8-TCDD
added, ng/g (ppb)
10.4
12.0
12.2
10.4
12.1
12.8
9.9
3.7
6.5
detected, ng/g (ppb)
10.6
8.4
11.0
9.7
8.4
10.1
9.24
1.38
5.6
Recovery, %
,102
70
90
93
69
79
94
37a
86
Portion of sample lost during preparation.
                                   33

-------
             TABLE  8.   RESULTS  OF  GC-MS-30  ANALYSES OF  SAMPLES
                      SPIKED WITH  37Cl4-2,3,7,8-TCDD
EPA
sample no.
C04130
C04131
C04132
5
6
4
12700
12701
11020
11025
WSU
sample no.
B-001C
B-002A
B-003A
B-006A
B-007A
B-008A
B-009E
B-010E
B-012F
B-017B
Quantity of
37Cl-2,3,7,8-TCDD
added, ng/g (ppb)
1.11
0.93
0.96
1.21
1.09
1.09
1.23
1.29
1.19
0.67
Quantity of
37Cl-2,3,7,8-TCDD
detected, ng/g (ppb)
0.78
0.91
0.61
0.48
0.67
0.75
1.06
1.14
0.93
0.58
Recovery, %
70
98
64
40
61
69
86
88
78
86
Data for samples 2 and 3 are not included because the ratio technique
could not be used with samples containing high levels of TCDD.  Sample
11024 is also omitted because the extract was not clean enough for
analysis by GC-MS-30.
                                   34

-------
                              m/e  321.8936
          m/e  319.8966

                I
Figure 8.  Dual-ion mass fragmentogram of sample 2
 obtained with GC-MS-30, mass resolution 1:12,500.
                        35

-------
                               m/e 321.8936
         m/e 319.8966
Figure 9.   Dual-ion mass fragmentogram of 150 pg of
   2,3,7,8-TCDD standard obtained with GC-MS-30,
             mass resolution 1:12,500.
                         36

-------
the ratio  of  m/e 319.8966 to m/e 321.8936 in the mass  fragmentogram is  0.79,
while  that for  the  2,3,7,8-TCDD  standard  is  0.84.   Both  of these values
agree well  with  the theoretically predicted ratio of  these  two  peaks,  £.77,
which is calculated on the basis of the  relative  abundance  of 35C1  and 37C1
isotopes.

     Further  confirmation  that  the  unknown component in sample 2 is indeed  a
quantity of TCDD isomers  is  provided  by  the observation that the  GC reten-
tion time  of  the unknown component  was identical  to  that of  the  2,3,7,8-TCDD
standard.   This  criterion  is  applied  in  all  determinations of TCDD's  in
Wright State's Brehm Laboratory.

     The mass spectrometric  resolution  achieved  in  this  program with  the
MS-30  Mass  Spectrometer  can  be demonstrated  experimentally by using  the
specialized  step-scan  circuitry developed by Wright  State.  The  practical
method of  demonstrating the  resolution is to obtain  a  narrow mass scan  for  a
sample consisting of  TCDD's  in a mixture  of  other compounds that yield mass
spectral  ions whose   mass is  very close  to that  of  TCDD's.   In  earlier
studies  we utilized a  mixture  of 2,3,7,8-TCDD,  PCB's such  as Aroclor  1254,
and DDE* for this  purpose.   The latter compounds yield mass  spectral  peaks
that  are  very  near  the  mass  of  the  TCDD's major  ion (Aroclor  1254  m/e
321.8679, DDE m/e 321.9290, 2,3,7,8-TCDD m/e 321.8936).

     In  order to obtain ions of approximately equal  intensity from all  these
compounds,   however, the quantities  of  PCB and DDE must be quite large  rela-
tive to  the quantity  of TCDD's.   Figure  10 shows  a  typical mass  fragmen-
togram  obtained  during  this  investigation  in analyses  of  two   mixtures  of
2,3,7,8-TCDD  and DDE and  a  mixture of Aroclor 1254,  2,3,7,8-TCDD,  and DDE.
On the basis  of  the data  shown  in  Figure 10, the dynamk: resolution of the
mass spectrometer is  calculated to  be  14,000 with 20  percent valley defini-
tion.

     The data on sample 2 which were  described  above  were based on monitor-
ing only m/e 320 and m/e 322  in the  mass spectrum  of  TCDD's.   Our earlier
experience   had shown  that the low levels  of TCDD's that are  usually found in
environmental samples  (low ppt) permit monitoring of  no more than four mass
peaks for  a single sample injection,  even with the sophisticated  step-scan
techniques   developed  in  Brehm  Laboratory.   In  this  instance,  however,  the
level  of TCDD's  (40  ppm) in sample 2 was very  high and it  was  feasible to
obtain an  actual mass  spectral  scan as  this component  of the sample eluted
from the gas chromatograph.

     Therefore the  MS-30  Mass Spectrometer was set up  in the normal magnetic
scanning mode, and  an aliquot  of the  extract of sample 2 was injected into
the GC.   At the  appropriate  retention  time,  the  mass  spectrum of the eluted
component  was  scanned.   Before  this,  we  obtained similar mass  spectra of  a
solution containing  10  ng  of  authentic  2,3,7,8-TCDD  standard  and  of  a
solvent  blank (benzene).    The instrumental  parameters applicable to  the
scans are as follows:
* As  previously noted,DDE  is a  degradation product of  the  pesticide DDT.


                                    37

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to
CO
                                                                             AROCLOR 1254
                                                                                                 lOOpg
                                                                                            .— 2,3,7,8-TCDD
                                                2,-3,7,8-TCDO
                                                               f-70ng  ODE
                           lOOpg
                       2,3,7,8-TCDD
                           m/e 319.8966x ^ m/e 319.9196
                                                  m/e 321.8936     m/e 321.9290
                                     NO. 1                  NO. 2
                                                                                                70ng  DDE
m/e 321.8936^  ^ m/e 321.9290
           NO. 3
                               Figure 10.   Mass fragmentograms  using GC-MS-30  of mixtures of
                                       2,3,7,8-TCDD  with  other chlorinated compounds.

-------
     Scan rate:
10 sec/decade,  beginning 190 sec.  after
 sample injection
     Mass range of scan: m/e 130 to m/e 350

     Mass resolution:     1:1000
     GC retention time
      for TCDD:
195 sec.
     Other parameters:    Same as described in Section 3

     The  relative  intensities  of  the more  prominent mass  spectral  peaks
recorded in these  runs  are listed in Table 9.   The mass  spectra obtained for
the  2,3,7,8-TCDD  standard  and  for  the  extract  of  sample 2  are shown  in
Figures  11  and 12.  These  spectra  obviously agree quite well.   There  is  no
doubt that the  unknown  component  in sample 2 is a TCDD isomer and  that  it is
present  in a  high concentration.   Apparently some  components of the  extract
of sample  2,  other  than  the TCDD's,  also contribute to  m/e   194, 257,  and
259,  but these are not of concern here.
                                    39

-------
TABLE 9.   RELATIVE INTENSITIES OF MAJOR IONS OBSERVED
                 IN MASS SPECTRAL SCANS
m/e
326
324
322
320
318
259
257
194
161
160
10 ng
2,3,7,8-TCDD
standard
10
50
100
80
30
23
34
18
21
17
Solvent blank
0
0
0
0
0
0
0
0
4
4
10 pi of
EPA sample 2 extract
(out of 2000 Ml total)
12
48
100
80
25
47
48
30
25
20
                          40

-------
ATTENUATION-
.1
m/e324 — '^
m/e 322

ATTENUATION
_uJ
1C
A

:
.
m/e 324--""^
m/e 322
)
^JL . A.VULU . J i . ^ , . i. , J
^^ m/e 318 m/e 259 '
m/e 320
J
J
MASS SPECTRUM OBTAINED FROM 10 ng of 2,3,7,8-TCDD STANDS
100 ATTENUATION: 10
. i . ..Lllil.i.l . -. 	 «i

jl
u
IR[

1
LL_ J
^ — m/e 318 m/e 259-^
m/e 320
, .
^ m/e 257
)

il.jjiL-,
^m/e 257
MASS SPECTRUM OBTAINED FOR A PORTION (5y£ OUT OF 10 m£ EXTRACT) OF SAMPLE 2
Figure 11.  « Mass spectra from scans of 2,3,7,8-TCDD standard and
            sample 2 (mass range m/e 330 to m/e 250).

-------
              ATTENUATION:  10
               J.1.1.  .,....   . .... I il
                                                                 ill   . .   .Jlllj.ll!
              m/e
                                  1                             JL
                                 194                       161^^160

     MASS  SPECTRUM  OBTAINED  FOR  A PORTION (5 y£ OUT OF 10 mi EXTRACT) OF  SAMPLE  2.
rv>
               ATTENUATION:   1
m/e
                                                       uL
                                                                       lull
                                                194                      161


                          MASS SPECTRUM OBTAINED FROM  10 ng of  2,3,7,8-TCDD  STANDARD
                       Figure 12.  Mass spectra from scans of 2,3,7,8-TCDD standard and

                                    sample 2 (mass range m/e 250 to m/e 150).

-------
                                  SECTION 5

                       CONCLUSIONS AND RECOMMENDATIONS
     As a  means of  assessing  the levels  of the extremely  toxic TCDD's  in
process streams, wastes,  and  sediments  from the manufacture of  chemicals, a
method was developed  that proved to  be  applicable to  about 70 percent of the
industrial  waste  sample  types  examined  in  this study.   These   sample  types
are typical  of those  that would be  collected  in  a  routine chemical  plant
survey.

     The analytical  methodology implemented  in  this  study  is summarized  in
the following five principal steps:

     I.   Preparation  of  a spiked and  nonspiked aliquot of each sample  in
          liquid extractable form (organic phase).

     2.   A sample clean-up procedure that includes  acid and base washes  to
          remove the bulk of the sample  matrix.

     3.   An additional  sample separation step using  liquid chromatography.

     4.   Screening of samples for  detectable  levels of-TCDD's with a  low-
          resolution  GC-QMS  system.   This  step  is   repeated with a spiked
          sample if positive levels  of TCDD's are detected.

     5.   Confirmation and quantification  of the level  of TCDD's by  analysis
          of the samples with a high-resolution GC-MS-30 system.

     There  are  four  major advantages with the implementation of  this  method:

     1.   The  procedure   offers  a relatively  rapid   method  for  qualitative
          screening of a wide  variety  of materials   for possible  contamina-
          tion by TCDD's,  through the use of low-resolution mass spectrome-
          try  (GC-QMS  showed  a  MDC  of  1  ppb or less  in  50 percent of the
          samples).

     2.   Only  samples  in which  the initial screening shows TCDD's  need  be
          confirmed  by  use  of  GC  with  high -resolution  mass   spectrometry
          (minimum resolution 1:10,000).

     3.   Analysis  by  high-resolution  mass  spectrometry yields  extremely
          high  sensitivity as  well  as  specificity.   The need  for both  is
          indicated  by  the finding of  minimum  detectable concentrations
          below 100 ppt in more than  half the samples  tested.
                                   43

-------
     4.   The  method warrants a high level of confidence owing to the use of
          an  internal  standard and  application  of the  four-ion monitoring
          technique.   Recovery  of  37Cl4-2,3,7,8-TCDD  from  spiked  samples
          indicates  a  recovery range of  40 to  98  percent for  the  method.
          Further,  by  a  procedure  in  which  the quantity  of  native-TCDD1s
          detected   is  proportionately  related   to  the  quantity of  37C14-
          2,3,7,8-TCDD  added,  the  data may  be   automatically  corrected  for
          recovery.

     Although  the procedures  outlined  here are  acceptable for  analysis  of
many  industrial  process   samples,   they  are  not applicable  to all  sample
types.    Among  those  examined  in this  study,  the samples  that  could  not  be
suitably  analyzed are of  two types.   First are  those of biological  origin,
primarily wood  and  woodlike products.  It  is  probable that for such samples
an  acid  digestion step is  needed to effectively destroy cellular walls  and
release  any  residue  of TCDD's.  Earlier work at Brehm Laboratory on wood and
other  biological  materials confirms  the effectiveness  of  such  an  approach.

     The  other  type of sample  not  amenable to the method  is  more  difficult
to  characterize.   Samples  of  this  type formed emulsions in  the preparation
phase  that  could not  be  resolved.   Use of several common  emulsion-breaking
techniques  such  as   addition   of  excess  solvent,   did  not  alleviate  this
problem.   Unfortunately,  owing to the small number of samples  of this type,
no  further information  was  obtained.   Additional  work on such  samples would
be desirable.
                                   44

-------
                                 REFERENCES
Albro,  P.  W. ,  and B.  J.  Corbett.   1977.  Extraction  and  Clean-up of Animal
Tissues  For Subsequent Determination  of Mixtures  of  Chlorinated Dibenzo-p-
dioxins and  Dibenzofurans.  Chemosphere, 7:381.

Baughman,  R. ,  and M.  Meselson.   1973a.  An  Analytical  Method for Detecting
TCDD  (Dioxin):   Levels  of TCDD  in  Samples  From Vietnam.   Environmental
Health Perspectives, 5:27.

Baughman,  R. ,  and  M.   Meselson.   1973b.   An  Improved  Analysis  for Tetra-
chlorodibenzo-p-dioxin.   In Chlorodioxins  -  Origin and Fate, E.  H.  Blair,
ed.    Advances  in  Chemistry,   Series  120,   American  Chemical  Society,
Washington,  D.C.

Bertoni,  G. ,  et  al.   1978.  Gas  Chromatographic Determination  of 2,3,7,8-
Tetrachlorodibenzodioxin  in the Experimental  Decontamination  of  Seveso Soil
by Ultraviolet Radiation.  Anal. Chem.,  50(6):732-735.

Blair,  E.  H. ,  ed.   1973.   Chlorodioxins  -   Origin  and  Fate.   Advances  in
Chemistry, Series 120, American Chemical Society, Washington,  D.C.

Buser,  H.  R.  1976.   High Resolution  Gas  Chromatography  of  Polychlorinated
Dibenzo-p-dioxins and Dibenzofurans.  Anal. Chem., 48:1553-1557.

Buser, H.  R.   1977.   Determination of 2,3,7,8-Tetrachlorodibenzo-p-dioxin in
Environmental  Samples   by  High-Resolution   Gas   Chromatography  and  Low-
Resolution Mass Spectrometry.  Anal. Chem., 49:918-922.

Buser, H.  R. ,  and H.  P.  Bosshardt.   1976.   Determination of  Polychlorinated
Dibenzo-pdioxins  and   Dibenzofurans   in  Commercial  Pentachlorophenols  by
Combined GC-MS.  Journal of the AOAC, 59(3):562.

Crummett,  W.  B. ,  and  R.  H.   Stehl.    1973.    Determination  of  Chlorinated
Dibenzo-p-dioxins  and  Dibenzofurans   in Various  Materials.   Environmental
Health Perspectives, 5:15.

Dow Chemical Co.   1980.  Science, in press.

Edmunds,  J.  W. ,  D.  F.  Lee,  and C.  M. L.  Nickels.    1973.   Pestic.  Sci.,
4:101.
                                   45

-------
 Elvidge,   D.   H.     1971.    The   Gas-chromatographic   Determination   of
 2,3,7,8-Tetrachlorodibenzo-p-dioxin   in   2,4,5-Trichlorophenoxyacetic   Acid
 (2,4,5-T),  2,4,5-T  Esters  and  2,4,5-Trichlorophenol.    Analyst  (London),
 96:721-727.

 Erk, S. D. ,  M.  L.  Taylor, and T.  0.  Tiernan.   1978.   Environmental  Monitor-
 ing in  Conjunction with  Incineration of Herbicide Orange at  Sea.   Proceed-
 ings of  the 4th National  Conference  and Exhibition on  Control  of  Hazardous
 Material Spills, pp.  226-231.

 Erk, S.   D. ,  M.  L.  Taylor,  and  T.  0.  Tiernan.    1979.   Determination  of
 2,3,7,8-Tetrachlorodibenzo-p-dioxin  Residues  on  Metal  Surfaces  by  GC-MS.
 Chemosphere, 8(1):7-14.

 Fee, D.  C.  ,  et al.    1975 .   Analytical  Methodology  for Herbicide  Orange
 Volume  II.   Determination of USAF  Stocks.   Aerospace Research  Laboratories
 Technical  Report TR75-0110 Volume II.

 Firestone, D. ,  et  al.   1972.   Determination  of  Polychlorodibenzo-p-dioxins
 and Related  Compounds   in Commercial  Chlorophenols.   Journal  of the  AOAC,
 55(l):85-92.

 Gross,  M.  L.   1978.   Personal  communication.

 Harless,   R.  L.   1976.    Presentation  given at TCDD  workshop  held  at  the
 Universita Di  Milano  Institute Di Farmacologia, Milano, Italy.

 Harless,  R.  L. and  A. Dupuy.   1979.   Personal communication.

 Hughes,  B.  M.,  et  al.   1975  .   Analytical  Methodology  for Herbicide  Orange
 Volume  I.   Determination  of Chemical  Composition.   Aerospace  Research  Labor-
 atories  Technical Report TR75-0110 Volume I.

 Lee,  D. ,  et  al.,  eds.    1973.    Environmental  Health  Perspectives, Experi-
 mental  Issue No.  5, September.

 McConnell, E.  E., J.  A. Moore,  J. K.  Haseman,  and  M.  W. Harris.  1978.   The
 Comparitive  Toxicity of  Chlorinated  Dibenzo-p-dioxins  in Mice  and  Guinea
 Pigs.  Toxicol.  Appl. Pharacol.,  44:335-356.

 Nestrick,  T. ,  L. Lampaski, and  R.  Stehl.   1979.   Synthesis and  Identifica-
 tion  of   the  22  Tetrachlorodibenzo-p-Dioxin  Isomers  by   High   Performance
 Liquid     Chromatography   and    Gas     Chromatography.      Anal.     Chem.,
 51(13):2273-2281.

 Nicholson,  W.  and  J.  Moore,  eds.   Health   Effects  of Halogenated Aromatic
 Hydrocarbons.  Annals of the New  York  Academy  of Science.   1979.  Vol.  320.

O'Keefe, P.  W. ,  M.  S. Meselson, and R. W. Baughman.  1978.  Neutral Clean-up
 Procedures  for  2,3,7,8-Tetrachlorodibenzo-p-Dioxin  Residues  in  Bovine  Fat
and Milk.  Journal of the  AOAC, 61:621-626.
                                   46

-------
Solch,  J.,  et  al.   1978.   Development  of GC-MS  Methodology  for  Assaying
Bovine  Tissue  for  Hexa-,  Hepta-,  and Octachlorodibenzo-p-dioxin  Content.
Proceedings  of  the  26th  Annual  Conference  on  Mass Spectrometry  and Allied
Topics, pp.  52-54.

Solch,  J., et al.   1979.   A  Unique Scan  Circuit  for Use  in  Multiple Ion,
GC-High Resolution  MS Determination of Picogram Quantities of Chlorodioxins.
Proceedings  of  the  27th  Annual  Conference  on  Mass Spectrometry  and Allied
Topics.  In  Press.

Taylor, M.  L.,  B.  M. Hughes, and T.  0. Tiernan.   1974a.   GC-MS Procedures
for  Characterization  of  Herbicide  Orange.   Disposal  of Herbicide  Orange,
USAF Environmental Health Laboratories, Brooks AFB, Texas.

Taylor, M.  L. ,  B.  M. Hughes, and T. 0. Tiernan.   1974b.  Preliminary Results
Obtained  with New  GC-MS Procedures  Developed  for  Determining Tetrachloro-
dibenzo-p-dioxin  in  Chlorophenoxy Herbicides.   Dioxin  Planning Conference,
EPA, Washington, D.C.

Taylor, M.  L.,  B.  M.  Hughes,  and T. 0.  Tiernan.   1974c.   Techniques  for
Analysis of  Dioxin in Chlorophenoxy Herbicides.  Status  of  Herbicide Orange
Disposition,  USAF  Environmental  Health  Laboratories,   Brooks  AFB,  Texas.

Taylor, M.  L.,  T.  0.  Tiernan,  and B. M.  Hughes.   1974.   USAF Analytical
Methodology   Developed  for  Characterization  of   Herbicide  Orange,  EPA,
Washington, D.C.

Taylor, M.  L.,  T.  0. Tiernan,  and B. M.  Hughes.   1975.   Analytical  Tech-
niques  for Determination of Chlorodioxins.   Proceedings,-1975 International
Controlled Release Pesticide Symposium, pp. 401-406.

Taylor,  M.   L.,  J.  G.  Solch,   and  T.   0.  Tiernan.    1979.   Advances  in
Analytical  Methodology  for  Ultratrace  Analysis  of  Tetrachlorodibenzo-p-
dioxin, Presented  at  the  Dioxin  Implementation  Plan  Collaborators Meeting,
January.

Taylor, M.  L.,  et  al.   1973.   Determination of Trace  Quantities of Chloro-
phenoxy-Type  Herbicides  and Related Chlorinated Residues in Soil Using GC-MS
Techniques.   Proceedings  of the  21st Annual Conference on Mass Spectrometry
and Allied Topics, pp. 336-339.

Taylor, M.  L.,  et  al.   1975.   Determination  of Tetrachlorodibenzo-p-dioxin
in  Chemical  and   Environmental  Matrices.    Proceedings  of  the  23rd Annual
Conference on Mass Spectrometry and Allied  Topics, pp.  337-340.

Taylor, M.  L.,  et  al.    1976.    Levels  of Tetrachlorodibenzo-p-dioxin  in
Environmental and   Biological  Samples  as  Determined by  Gas Chromatography-
High  Resolution  Mass Spectrometry.   Proceedings  of   the  24th  Annual  Con-
ference on Mass Spectrometry and Allied Topics, p. 595.
                                   47

-------
 Tiernan,  T.  0.   1975b.   Applications of  Mass  Spectrometric Techniques  for
 Pesticide Characterization,  and Monitoring Environmental Effects.   Interna-
 tional  Controlled Release Pesticide Symposium.

 Tiernan,  T.  0., M.  L.  Taylor, and  B.   M.  Hughes.   1975.   Measurement  of
 Tetrachlorodibenzo-p-dioxins in  USAF  Herbicide Stocks  and  in  Environmental
 Samples.    Sixth   Annual   Symposium  on   Environmental  Research,   Edgewood
 Arsenal,  Maryland.

 Tiernan,  T.  0.,  et al.   1979.   Methodology  for  Gas  Chromatographic-High
 Resolution   Mass Spectrometric   Determination  of  Chlorodioxins  in  Complex
 Samples,  llth Ohio Valley Chromatography Symposium.

 Webber,  T.   J.   N.,  and  D.  G.   Box.   1973.  The  Examination of  Tetrachlor-
 vinphos   and  its   Formulations   for  the  Presence  of  Tetrachlorodibenzo-p-
 dioxins  by  a  Gas-Liquid  Chromatographic  Method.   Analyst (London),  98:181.

 Williams,  D. T., and B.  J.  Blanchfield.   1971.   Thin  Layer Chromatographic
 Separation   of   Two   Chlorodibenzo-p-dioxins   From   Some   Polychlorinated
 Biphenyls and  Organochlorine Pesticides.   Journal of  the AOAC,  55:1429-1431.

 Williams,  D.  T.,  and  B.  J. Blanchfield.   1972.   Screening Method for  the
 Detection  of  Chlorodibenzo-p-dioxins  in  the  Presence  of   Chlorobiphenyls,
 Chloronaphthalenes,  and Chlorodibenzofurans.   Journal of the AOAC,  55:93-95.

 World  Health Organization,  1977.  IARC Monograph on  the Evaluation of Car-
 cinogenic  Risk of  Chemicals to  Man.   Some Fumigants,  the  Herbicides 2,4-D
 and   2,4,5-T,    Chlorinated  Dibenzodioxins  and   Miscellaneous  Industrial
 Chemicals.   Lyon, France,  Vol. 15,  August.

 Wright  State University.   1976.   Annual   Report  on  U.S.  EPA  Contract  No.
 68-01-1959.   The Analysis  of Environmental Samples  for TCDD Utilizing High
 and  Low  Resolution  Gas-Liquid  Chromatograph-Mass Spectrometry.   EPA Office
 of Special Pesticide  Reviews, Washington,  D.C.

 Yelton,  R.   0.,  M.   L.   Taylor,   and  T.  0.  Tiernan.   1977.   Ultrasensitive
 Method for  Determination  of  Chlorodioxins  in  Commercially Produced  Phenols.
 Proceedings  for  the  25th  Annual Conference on Mass  Spectrometry and Allied
Topics, p. 595.
                                   48

-------
                                 APPENDIX A
         BASIC PRINCIPLES OF GAS CHROMATOGRAPHY, MASS SPECTROMETRY,
                            AND COMBINED SYSTEMS
GAS CHROMATOGRAPHY (GC)

     Gas chromatography  is  a special  form of  chromatography  that  is used to
separate the  components  of chemical mixtures.   Several  excellent  references
describe the  technique  in  detail  (Dal  Nogare and  Juvet 1962;  Littlewood
1970; Jones 1970;  Ambrose 1971).   In gas chromatography  the  mobile phase is
a  gas  and  the  stationary phase  is  either a  liquid or  a solid,   hence  the
terms  gas-liquid  chromatography  and  gas-solid chromatography.   Gas-liquid
chromatography  entails  the  use of a  separation device,  which is  a  column
containing  the  liquid  phase  (typically a   high-boiling  organic  silicone
polymer) distributed  on  a  highly  inert solid support.   Figure Al  depicts  a
typical gas chromatograph.

     The column  is maintained  in  an  oven,  in which the  temperature  can be
controlled precisely;  through the  column is  passed an inert,  high-purity gas
(e.g.,  helium),  called the carrier gas.  The carrier gas is the mobile phase
and  the organic  silicone  polymer  is  the   liquid  phase.   Typically,  the
samples are introduced into the column in 0.1 to 10 ul  amounts with a micro-
syringe through  an injection port, which is  a heated (100°  to 250°C)  inlet
system equipped  with  a silicone septum.  The sample is  vaporized immediately
upon injection,  and  The  inert carrier gas passing through the injection port
sweeps  the  volatilized,   injected  sample  out  of the  injection  port and into
the gas chromatographic  column.   The volatilized constituents  of  the sample
migrate through  the  column at varying rates  because  of variations  in  the
physical and  chemical properties  of  each component, such  as boiling point,
absorptivity,  and  solubility.   The components are  thus  separated  and emerge
(elute) from  the column  at different times.    In  some samples the  components
are highly  similar and are not effectively separated or may necessitate the
use  of extraordinary  chromatographic  procedures.   More  commonly, however,
the  components   of  a  chemical  mixture  can  readily  be separated  by fairly
simple  gas  chromatographic techniques.

     As each  separated component  elutes from the gas chromatographic column,
it  is  detected  by one  or more of  several  types  of detectors.   Among  the
widely  used detectors  are flame ionization,  thermal  conductivity,  and elec-
tron capture  detectors.   Other,  more specific, types of  detectors are also
used in  conjunction with  gas chromatography;  in particular,  the  mass spec-
trometer has  been  used extensively.   A discussion  of the principles of mass
spectrometry follows.
                                     49

-------
               FINE ADJUSTMENT
                    VALVE
      DRYING TUBE
KX1
9                                     MANOMETER OR
                                    PRESSURE GAUGE
                  SAMPLE  INJECTOR-
/"S PRESSURE
yj REGULATOR
     CYLINDER CONTAINING
        CARRIER GAS
FLOWMETER
                                             DETECTOR
                                                               COLUMN
                                             THERMOSTAT
              Figure Al.   Apparatus for gas chromatography
                                   50

-------
MASS SPECTROMETRY (MS)

     Mass spectrometry  is  described  in  detail  in several references  (Beynon
1960; McLafferty  (ed.)  1963;  Kiser 1965;  Roboz  1968;  McFadden  1973).  Figure
A2  is  a  schematic diagram  of  a  typical mass  spectrometer;  the  principal
components of  such  a  system  are (1) an inlet system  (2) an ion source,  (3)
an  accelerating  system, (4)  an analyzer  system,  (5) a detector,  and (6) a
data acquisition  system.   The  functions  of these  components  are  described
briefly.

     The  inlet system  is  the means of introducing the sample into  the  ion
source of the  mass  spectrometer.   Inlet devices in  common use  include heated
direct insertion  probes and  heated  gas inlet  systems  (batch  inlets), which
are coupled to the  mass spectrometer through a  restricted  fixed or variable
orifice,   often called  a "leak."   In  recent  years the  gas  chromatograph  has
been  used  often  to   introduce  the  sample and  is  coupled  to  the  mass
spectrometer—hence the term "coupled GC-MS."

     Because   the   ion   source,   the accelerating   lens system,   the  mass
analyzer, and  the  detector of the mass  spectrometer are all maintained  under
vacuum by a pumping system,  the inlet  system must  admit the sample (and  the
carrier  gas  of a  gas chromatograph) into  the  spectrometer at  such a rate
that the  pumping  system maintains the specified  internal operating pressure
of the instrument.

     The  ion  source  (shown  schematically in Figure  A3) is typically  main-
tained at pressures of  10  3mm and lower (10  6mm) and  at temperatures of 100°
to  250°C.   The source  is  the  region  in  which  ions  are generated  from  the
volatile sample molecules  admitted through  the  inlet  system.   The  ionization
of molecules in the gas phase  is  effected by bombarding them  with electrons
emitted  from a hot metal wire  or  ribbon  (the  filament) and drawn  through a
set of slits for  collection  at an anode or  electron trap.   The energy of  the
electrons is controlled by the potential  difference between the filament  and
the trap.   As  these  energetic  electrons  either strike or  pass close to  the
sample molecules,  ionization  occurs,  producing  a molecular  ion that usually
is  fragmented  further to yield  other ions  of  smaller  mass.   The  ion source
produces both  positively charged and negatively charged ions,  and many mass
spectrometers  in use today are designed to detect both types.

     The  ions  produced  are  electrically  forced  out  of  the ion  source  and
into the  accelerating lens system, which  generally imparts  several kilovolts
of energy to the ions, which then enter the mass analyzer section.

     The  purpose  of  the mass  spectrometer  analyzer  is  to  separate the  ions
according to  their mass:charge  ratios.   Various  types of  analyzer systems
are in use  today,  and the type  of analyzer  usually provides the  descriptive
name  for each mass  spectrometer  system.    Thus  there are,  for  example,
quadrupole  mass  spectrometers,  single-focusing  magnetic  deflection  mass
spectrometers,   time-of-flight mass  spectrometers,  and  double-focusing  mass
spectrometers.   Each  of these systems is  characterized by a distinct mode of
ion separation, and each provides different capabilities.
                                     51

-------
         SAMPLE
        RESERVOIR
INLET
                         TO  VACUUM
                                                                 OSCILLOGRAPH
 MOLECULAR
   LEAKS
    ION SOURCE
IONIZING ELECTRON BEAM

  ACCELERATING REGION
                                                                     RESOLVING
                                                                       SLIT
                    ANALYZER
                      TUBE
                                     RESONANT
                                     ION BEAM
                                            MAGNET
Figure A2.   Schematic diagram  of a Nier 60° sector mass spectrometer.
                                      52

-------
en
CO
                IONIZATION CHAMBER-
                                         •REPELLER
                                               ^FILAMENT
r-ELECTRON SLIT

     r-FIRST ACCELERATING
       SLIT       (-SECOND ACCELERATING SLIT
                                                   '   IONIZING  REGION
                  MOLECULAR LEAK-J
                            ELECTRON  BEAM -I
                                           ANODE —'
       ION ACCELERATING
            REGION
                     Figure A3.  Electron-impact ion source and ion accelerating system.
                                      Source:  Merritt and Dean   1974.

-------
      The ability  of a mass  spectrometer  to  effect a separation of adjacent
 mass peaks  (that  is,  to  resolve these  peaks) depends  upon  the analyzer.
 Resolution is defined  by  the equation,  R = M/AM,  where M  is the mass of the
 first peak  in a  doublet  and AM is the difference  in  the  masses  of the two
 peaks.   An  increase in the  value  of  R  (denoting an increase in resolution)
 indicates  an  increase  in the  ability  to  distinguish between  very nearly
 identical  masses.   Of  the  several  mass  spectrometers mentioned, the double-
 focusing  type  affords the   greatest  mass  spectral  resolution,  sometimes
 exceeding  100,000.   At this  degree  of  resolution,  masses  appearing at m/e
 99,999  and  m/e  100,000 would  be  distinguishable.   An instrument capable of
 such high  resolution is of course very  complex and  expensive and thus would
 be used only  when  such high  resolution  is mandatory for effective analysis.
 In contrast,  a  quadrupole mass spectrometer  is  much simpler to operate and
 less expensive  but  can  provide  only low  resolution (m/Am  = 500  to  1000
 typically).

      Detection of  the   ions  that  have  been  separated is  accomplished  most
 often by use  of  an electron multiplier,  of which,  again,  various  types are
 in use.   An electron multiplier produces  current amplification of 103 to 108
 with very  low  noise  level   and   with  negligible  time  constant or  signal
 broadening.   The  amplified analog  signal  resulting  from the ion impacting on
 the  electron  multiplier  is  finally  routed  to one  of  several  possible  data
 acquisition  devices; among those often used  are  the ocillographic recorder,
 the  analog recorder,  a  pulse  counting  device,  or  the digital computer.

      The  data from  a mass spectrometer  consist, in  the analog  format,  of a
 spectrum  of  peaks  (the mass  spectrum).   The position of  each peak  on the
 horizontal  axis  of a  graphic  display indicates  its m/e  ratio whereas the
 amplitude  of  each  peak indicates  the number of  ions  (or  abundance)  of  that
 m/e.  The data may  also be  displayed digitally  in tabular form.

      If  more  than one  compound enters the mass spectrometer at a given time,
 then  the masses  detected  are  generally  attributable to any  or all  of the
 compounds.    Because it  is  difficult,  and  sometimes impossible,  to interpret
 the  mass spectra  obtained for  mixtures of organic compounds, there is great
 advantage  in  admitting  the compounds separately.   Thus  a  gas chromatograph
 is used  to  introduce the separated components of a mixture sequentially into
 the  mass  spectrometer.   Following  is a simplified  description  of  a coupled
 GC-MS system.


GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC-MS) SYSTEMS

     In  considering the coupling  of  the  gas  chromatograph to  a  mass spec-
 trometer, one  should recall  that  the source, analyzer, and  detector  of the
 spectrometer   are   all   typically   maintained  at  pressures  below  10  5mm.
Therefore,   unless  the  mass   spectrometer is  equipped  with  a very  high-
capacity pumping  system,  the  gaseous effluent  from a gas  chromatographic
column cannot  be  admitted  directly to the mass  spectrometer  source because
this would increase the pressure to a level  that would prevent satisfactory
operation.   Therefore,  coupling is  generally achieved by  use  of  an inter-
                                     54

-------
mediate  device  to  reduce the  rate  of flow  of the  sample  and carrier  gas
stream.  For  this  purpose several  types of devices (called "separators1,1)  are
used  to achieve  partial  separation  of the  carrier  gas  (typically  helium)
from  the gaseous  sample molecules.   Among  these devices are  (1) a  porous
barrier  or  effluent  splitter,  (2)  a jet/orifice  separator,  and  (3)  a
molecular separator  that includes  a  permeable membrane.   Some  gas  chromato-
graph/mass spectrometer  systems  feature a  direct coupling of the  gas chrom-
atograph with the  mass spectrometer  by means of a very  high  capacity  pumping
system.

     A  system that  couples  a  chromatograph  with a  mass  spectrometer is  a
very  powerful  analytical  tool,  the  only system that can  provide  definitive
analysis of  complex chemical  mixtures.   The separation capabilities  of  the
gas  chromatograph  are complimented  by the  inherent  specificity and  sensi-
tivity of the mass spectrometer.   During analysis of a complex mixture,  the
components are separated gas  chromatographically, each  eluted component then
passes  through  the  interface  (separator)  and  into  the  mass  spectrometer,
which  provides  and  records  a mass  spectrum.   Typically,  the  analysis of  a
mixture  could yield  several  hundred  mass spectra, each  containing  100 to  200
mass peaks.    Therefore,  the  computer is an ideal means  of acquiring the mass
spectra,  reducing  the  data   (converting  the  acquired  data  to actual  mass
spectra  by  comparison with  calibrated  reference files),  and displaying  the
data.   The  minicomputer  is an  essential  component of  a modern GC/MS system
because  the  analyses  generate such  sizable  quantities of  data.   Use of  a
minicomputer  can  afford  other  advantages;  for example, the computer can be
programmed  to  control  the  mass  spectrometer  so   that  it  monitors  only
selected masses typical  of the compounds of interest.   The computer also can
be  programmed to  allow monitoring   of different masses- (corresponding  to
different compounds) at different gas chromatographic retention times.
                                     55

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                                  APPENDIX A

                                  REFERENCES


 Ambrose,  D.   1971.   Gas  Chromatography.   Van Nostrand,  New York.

 Beynon,  J.  H.    1960.   Mass  Spectrometry  and  Its  Applications  to  Organic
 Chemistry.   Elsevier,  Amsterdam.

 Dal  Nogare,  S.   and  R.   S.  Juvet.   1962.  Gas  Chromatography:   Theory  and
 Practice.   Wiley-Interscience,  New York.

 Jones,  R.  A.  1970.  An  Introduction  to  Gas-Liquid Chromatography.   Academic
 Press, New York.

 Kiser,  R.  W.  1965.  Introduction  to  Mass Spectrometry and Its Applications.
 Prentice-Hall, Englewood  Cliffs,  New Jersey.

 Littlewood,  A.  B.  1970.  Gas  Chromatography.   2nd Ed., Academic  Press,  New
 York.

 McFadden,  W.  1973.   Techniques  of Combined  Gas  Chromatography/Mass  Spec-
 trometry.  Wiley-Interscience,  New  York.

 McLafferty,  F. W. ,  ed.   1963.  Mass Spectrometry of Organic  Ions.   Academic
 Press, New York.

 Merritt, W.  H. ,  Jr. , and J.  Dean.   1974.   Instrumental Methods of Analysis.
 5th Ed.  Van Nostrand, New York.

 Roboz,  J.    1968.   Introduction  to Mass Spectrometry.  Wiley-Interscience,
New York.
                                     56

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                                 APPENDIX B

               OTHER INSTRUMENTAL METHODS FOR DIOXIN ANALYSIS
     Most of  the  current technology for detection of  TCDD's  is  based  on  gas
chromatography  and/or  mass spectrometry.   However,  a variety of  other less
specific  techniques   have   been   used  including  ultraviolet  spectroscopy
(Pohland  and  Yang 1972),  electron spin  resonance  spectroscopy,  and low-
temperature  phosphorescence  emission spectroscopy (Baughman 1974).  None  of
these methods provide both  the  high sensitivity and  selectivity  needed  for
analysis of most environmental samples.

     A  resin  sorption technique  using XAD-2 resin has achieved a detection
limit of 1  ppt for TCDD's in water; because this technique required a large
quantity of  sample  for  extraction,  however,  extension  to  other types  of
samples is unlikely (Junk 1976).

     Another  technique  uses  PX21  powdered  charcoal   suspended  on  shredded
polyurethane  foam  as   the  sorbant (Huckins, Stalling, and  Smith 1978).   The
TCDD's were  eluted from the charcoal column  by  use  of a  50 percent solution
of  toluene   in  benzene  and  finally were  detected by electron-capture  gas
chromatography.   To enhance  selectivity,   an alumina column  chromatography
step is usually included after elution from the charcoal  column.   The  detec-
tion limit of this method ranges from 10 to 100 ppb.

     Thin-layer  chromatography  has also  been  used  for  the detection  of
TCDD's  (Williams   and  Blanchfield  1971).   Two-dimensional development with
two  different  solvents  is  used  to increase  selectivity.   The spot  corre-
sponding to  2,3,7,8-TCDD is  removed from the  plate,  extracted with benzene,
and  detected  by  electron -capture gas chromatography.   This  method  has
achieved a detection limit in the low ppm region.

     Steam distillation  has  also  been  tried (Storhen  1971), but was suitable
only for levels of TCDD's in the range  of  1 to 3 ppm and lacked  the  selec-
tivity needed to avoid interferences.

     Recently  analytical methods  involving  chemical  ionization  mass  spec-
trometry with  negative ions  have been  published.  An  early communication by
Hunt and  co-workers (Hunt,  Harvey, and Russel  1975) reported  a  signal-to-
noise ratio  of  50  from a 2-pg direct-probe  insertion sample using oxygen as
the reagent  gas.   A sensitivity 25  times higher than  the  direct-probe  inser-
tion method  is  reported for electron impact ionization.   Hass et al.  compare
the  relative  sensitivities of  various chemical  ionization modes, including
those of  positive-ion  versus  negative-ion modes  with methane, oxygen,  and
                                     57

-------
mixed  methane/oxygen  as  reagent gases  (Mass  1978).   Positive-ion  chemical
ionization  affords  the  greater  sensitivity,   but  does  not  produce  ions
indicative of the molecular weight.
                                    58

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                                 APPENDIX B
                                 REFERENCES
Baughman, R. W.  1974.  Ph.D. Thesis, Harvard  University,  Cambridge,
     Massachusetts.

Mass, J. R., et al.  1978.  Anal. Chem.,  50:1474.

Huckins, J. N., D. L. Stalling,  and W.  A.  Smith.   1978.   Journal  of the
     AOAC,  61:32.

Hunt, D. F., T. M. Harvey, and J. W.  Russel.   1975.   J.C.S.  Chem.  Comm.,
     Vol. 151.

Junk, G. A., et al.  1976.  J. Am. Water  Works Assoc.,  68-218.

Pohland, A. E., and G. C. Yang.   1972.  J.  Agric.  Food  Chem.,  20:1093.

Storhen, R. W., et al.  1971.  Journal  of the  AOAC,  54:218.

Williams, D. T., and B. J. Blanchfield.   1971.   Journal  of the AOAC,
     55:93-95.
                                      59

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                                  APPENDIX C

                               LITERATURE REVIEW

      This appendix is a compilation of references on dioxin analysis cate-
 gorized by sample matrix.   The categories are given below:

      Air                           Hexachlorobenzene

      Biological  tissue             Insecticides

      Blood                         Milk or cream

      Commercial  chlorophenols      Plant material

      Fats or oils                  Soil

      Fish and crustaceans           Urine

      Flue Gas                      Water

      Fly ash                       Wipe samples

      Grain                         Wood

      Herbicide formulations

Air

Oswald,  E.   1979.  Toxicology Research Projects Directory,  Vol.  04,  Iss.  07.

Biological Tissue

Baughman,  R., and M.  Meselson.   1973.   Environmental  Health Perspectives,
      5:27.

Bradlaw,  J.  A., et al.  1975.  Proceedings  of Society of  Toxicology  Meeting,
     Williamsburg, Virginia,  March.

Freudenthal, J.   1978.  In:   Dioxin:   Toxicological  and Chemical  Aspects,  F.
     Cattabeni, A. Cavallaro,  and G. Galli, eds.   Spectrum  Publications,
      Inc., New York,  Chapter  5:43-50.

Hass, J.  R., et al.   1978.  Anal. Chem.  Vol.  50.

McKinney, J.  D.   1978.  In:   Chlorinated  Phenoxy  Acids and  Their  Dioxins.
     Ecol. Bull., 27:53-66.

                                     60

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0-Keefe, P. W.  1978.  In:  Dioxin:  Toxicological  and  Chemical  Aspects.   F.
     Cattabenl, A. Cavallero, and G. Galli,  eds.  Spectrum  Publications,
     Inc., New York, Chapter 7:59-78.

Oswald, E.  1979.  Toxicology Research Projects  Directory,  Vol.  04,  Iss.  07.

Rose, J. Q., et al.  1976.  Toxicol. Appl. Pharmacol.,  36:209.

Shadoff, L. A., and R. A. Hummel.   1978.  Biomed Mass Spectrom,  5(1):7-13,
     January.

Tiernan, T. 0.  1976.  EPA Contract No. 68-01-1959.  December.

Woolson, E. A., R. F. Thomas, and P. D. J. Ensor.   1972.   J.  Agric.  Food
     Chem., 20:351.

Woolson, E. A., et al.   1973.  Advanced Chemistry Series.

Young, A.  L.  1974.  Report No.  AFATL-TR-74-12,  Air Force  Armament
     Laboratory,  Eglin Air Force Base, Florida.

Blood

Hummel, R.A.  1977.  J.  Agric. Food Chem., 25:1049-1053.

Oswald, E.  1979.  Toxicology Research Projects  Directory,  Vol.  04,  Iss.  07.

Commercial Chlorophenols

Blaser, W. W., et al.  1976.  Anal. Chem., 48:984.

Buser, J.  R.  1975.  J.  Chromatography, 107:295.

Buser, J.  R., and H. P.  Bosshardt.  1976.  Journal  of  the  AOAC,  59:562.

Crummet, W. B., and R. H. Stehl.  1973.   Environmental  Health Perspectives,
     5:15.

Firestone, D., et al.  1972.  Journal of  the AOAC,  55:85.

Higginbotham, G.  R., et  al.  1968.  Nature  (London), 220:702.

Lamberton, J., et al.  1979.  J. Amer. Ind.  Hyg.  Assoc.,  40:816-822.

Langer, H. G., et al.  1971.  162nd Meeting, ACS, Washington, D.C.,  Pest.
     Sec., No. 83.

Micure, J. P., et al.  1977.  J. Chromatogr. Sci.,  7:275.
                                     61

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 Pfeiffer, C.   1976.  J. Chromatogr. Sci., 14:386.

 Pfeiffer, C.  D.,  T. J.  Nestrick, and C. W. Kocher.  1978.  Anal. Chem.,
      6:800.

 Fats or Oils

 Campbell, J.  C. ,  and L. Friedman.  1966.  Journal of the AOAC, 49:824.

 Firestone, D.   1976.  Journal of the AOAC, 59:323-325.

 Firestone, D.   1977.  Journal of the AOAC, 60:354-356.

 Higginbotham,  G.  R., et al.   1967.   Journal of the AOAC, 50:874.

 Horwitz,  W.,  ed.   1975.  Official Methods of Analysis of the Association of
      Official  Analytical  Chemists,  Association of Official Analytical
      Chemists, Washington, D.C., 12th Ed., Sect.  28.118, pp.  511-512.

 Hummel,  R. A.   1977.   J.  Agric.  Food Chem., 25:1049-1053.

 Kocher,  C. W., et al.   1978.   Bulletin of Environmental Contamination and
      Toxicology,  19:229.

 O'Keefe,  P. W., M.  S.  Meselson,  and R.  W.  Baughman.   1978.  Journal of the
      AOAC, 61:621-626.

 Ress, J.  R., G. R.  Higginbotham, and D. Firestone.  1970.  Journal  of the
      AOAC, 53:628-634.

 Shadoff,  L. A., et  al.   1977.  Annali  di Chimica, 67:583.

 Shadoff,  L. A., and R.  A.  Hummel.   1978.   Bio.  Mass  Spec., 5:7.

 Williams,  D. T.,  and B. J. Blanchfield.  1971.   Journal of the AOAC,
      54:1429-1431.

 Williams,  D. T.,  and B. J. Blanchfield.  1972.   Journal of the AOAC,
      55:93-95.

Williams,  D. T.,  and B. J. Blanchfield.  1972.   Journal of the AOAC,
      55:1358-1359.

 Fish  and Crustaceans

Baughman, R.  W.,  and M. Meselson.   1973.   166th Nat.  Meeting,  ACS,
      Chicago,   Abstract  Pest.,  55.

Baughman, R.  W.,  and M. Meselson.   1973.   Environmental Health Perspectives,
      Expt.  Issue 5, 27-35.
                                     62

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Baughman, R. W.  1974.  Ph.D. Thesis, Harvard  University,  Cambridge,
     Massachusetts.

Fukuhara, K., etal.  1975.  J. of Hvg. Chem.,  21:318.

Gross, M. L.  1978.  Personal communication, November.

Lamparski, L. L., T. J. Nestrick, and R.  H.  Stehl.   Anal.  Chem.,
     51(9):1453-1458.

Shadoff, L. A.,  and R. A. Hummel.  1975.   170th Nat.  Am.  Chem.  Soc.  Meeting,
     Chicago, Ab. Anal., Vol. 80.

Shadoff, L. A.,  et al.  Bull. Environ. Contain.  Toxicol.   In press.

Flue Gas

Frigerio, A., and M. C. Tagliabue.   Impianti  Incenerimento Rifuite Solidi:
     Prelievo, Anal. Controllo  Effluenti,  [conv.];  59-71.

Fly Ash

Buser, H. R., H. P. Bosshardt,  and C. Rappe.   1978.   Chemosphere,  2:165.

Grain

Hummel, R. A.  1977.  J. Agric. Food Chem.,  25:1053-1099

Isensee, A. R.,  and and G. E. Jones.  1971.  J.  Agric.  Food Chem., 19:1210.

Shadoff, L. A.,  and R. A. Hummel.  1978.   Biomed Mass  Spectrom,  5(1):7-13,
     January.

Herbicide Formulations

Brenner, K. S.,  K. Muller, and  P. Sattel.  1972.  J.  Chromatography, 64:39.

Brenner, K. S.,  K. Muller, and  P. Sattel.  1974.  J.  Chromatography,
     90:382-387.

Buser, H. R., and H. P. Bosshardt.   1974.  J.  Chromatography, 90:71.

Crummett, W. B., and R. H. Stehl.  1973.   Environmental  Health Perspectives,
     5:15.

Edmunds, J. W.,  D. F. Lee, and  C. M. L. Nickels.  1973.   Pestic.  Sci.,
     4:101.

Elvidge, D. A.   1971.  Analyst  (London),  96:721.
                                      63

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 Hackins, J. N., D. L. Stalling, and W. A. Smith.  1978.  Journal  of  the
      AOAC, 61:32.

 Hohmstedt, B.  1978.   In:  Dioxin:  Toxicological and Chemical Aspects.   F.
      Cattabeni, A. Cavallaro, and G. Galli, eds.  Spectrum Publications,
      Inc., New York,  Chapter 3:13-25.

 Hughes,  B. M., et al.  1975.   Natl. Tech. Inform. Serv.,  AD-A011, 597:Vol.
      1.

 Polyhofer, K.  1979.   Levensm Unters Forsch.  168(1):21-4, January.

 Ranstad, T.,  N.  H. Mahle, and R. Matalon.  1977.  Anal. Chem., 49:386.

 Rappe, C., H. R.  Buser,  and H.  P.  Bosshardt.  1978.   Chemosphere, 5:431.

 Shadoff, L.  A.,  etal.   1978.  Anal. Chem., 50(11):1586-1588.

 Tiernan, T.  0.   1976.  EPA Contract No. 68-01-1959,  December.

 Tiernan, T.  0.,  M. L. Taylor, and B. M. Hughes.   1975.  Proceedings  1975
      International Controlled Release Pesticide Symposium.

 Vogel, H.,  and R.  D.  Weeren.   1976.   Anal.  Chem., 280:9.

 Woolson, E. A.,  R.  F. Thomas, and P. D. Ensor.  1972.  J. Agric.   Food Chem.,
      20:351.

 Hexachlorobenzene

 Villanueva, E.  C., et al.   1974.   J. Agric.  Food Chem., 22:916.

 Insecticides

 Elvidge, D. A.  1971.  Analyst,  96:721.

 Storherr,  R.  W., et al.   1971.   Journal of  the AOAC,  54:218.

Webber,  T. J. N.,  and D.  J. Box.   1973.  Analyst (London),  98:181.

Woolson,  E. A., R.  F. Thomas, and  P.  D. J.  Ensor.   1972.  J.  Agric.   Food
     Chem., 20:351.

Shadoff,  L. A., et al.  Bull. Environ.  Contam. Toxicol.   In press.

Shadoff,  L. A., and R. A.  Hummel.   1978.  Biomed Mass Spectrom, 5(1):7-13
     January.
                                      64

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Milk or Cream

Baughman, R., and M. Meselson.  1973.   Environmental  Health  Perspectives,
     Exp.   Issue 5, 27-35.

Baughman, R. W.  1974.  Ph.D. Thesis, Harvard  University,  Cambridge,
     Massachusetts.

Hummel, R.  A.  1977.  J. Agric. Food Chem.,  25:1049-1053.

Plant Materials

Buser, H. R.  1977.  Anal. Chem., 49:918.

Buser, H. R.  1978.  Monogr. Giovanni Lorenzini  Found.;  Vol  1,  In:   Dioxin:
     lexicological and Chemical Aspects,  27-41.

Di Domenico, A., et al.  1979.  Anal Chem;  51(6):735-740.

Hummel, R.  A.  1977.  J. Agric. Food Chem.,  25:1049-1053.

Shadoff, L. A., and R. A. Hummel.  Biomed Mass Spectrom, 5(1):7-13,  January.

Soil

Bertoni, G. , et al.  1978.  Anal. Chem.,  6:732.

Buser, H. R.  1977.  Anal. Chem., 49:918.

Buser, H. R.  1978.  Monogr. Giovanni Lorenzini  Found.;  Vol  1,  In:   Dioxin:
     lexicological and Chemical Aspects,  27-41.

Camoni, I.  1978.  J. of Chromatography,  153:233-238.

Di Domenico, A., et al.  1979.  Anal Chem.,  51(6):735-740.

Gross, M. L.  1978.  Personal communication, November.

Hummel, R.  A.  1977.  J. Agric. Food Chem.,  25:1049-1053.

Kearney, P. C., E. A. Woolson, and C. P.  Ellington.   1972.   Environ.  Sci.
     Technol., 1017.

Nash, R. G.  1973.  Journal of the AOAC,  56:728.

Shadoff, L. A., and R. A. Hummel.  1975.   170th  National American Chemical
     Society Meeting, Chicago, Illinois,   Abst.  Anal.,  80.

Shadoff, L. A., et al.  Bull. Environ.  Contain. Toxicol.   In  press.
                                     65

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 Shadoff,  L.  A.,  and R.  A.  Hummel.   1978.   Biomed Mass Spectrom, 5(1):7-13,
      January.

 Widmark,  G.   1971.   Tracer Cosmos, a Realistic Concept in Pollution
      Analysis.   In:   International Symposium on Identification and
      Measurement of Environmental  Pollutants, B.  Westley, ed.  National
      Research Council  of Canada,  Ottawa,  p.  396.

 Woolson,  E.  A.,  et  al.   1973.'  Advances in Chemistry Series, 120:112.

 Urine

 Oswald, E.   1979.   Toxicology Research Projects Directory, Vol. 04, Iss.  07.

 Water

 Junk, G.  A.,  et  al.   1976.   J.  Am.  Water  Works Assoc., 68:218.

 Shadoff,  L.  A.,  and  R.  A.  Hummel.   1978.   Biomed Mass Spectrom, 5(1):7-13,
      January.

 Wong, A.  1978.   EPA Contract No.  68-03-2678, July.

 Wipe  Samples

 Di Domenico, A.,  et  al.  1979.  Anal.  Chem.,  51(6):735-740.

 Erk,  S.  D., M. L. Taylor,  and T. 0.  Tiernan.   1979.   Chemosphere,  8(1):7-14.

Wood

Hass, J. R., et  al.   1978.   Anal.  Chem.,  50:1474.

 Levin, J.  D., and C. A.  Nilsson.   1977.   Chemosphere,  7:443.
                                     66

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                                   TECHNICAL REPORT DATA
                            (Please read Instructions on the reverse before completing)
1. REPORT NO.
   EPA-600/2-80-157
                              2.
                                                           3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE
  Dioxins:  Volume  II. Analytical  Method
            For Industrial  Wastes
             5. REPORT DATE
                JUNE 1980 ISSUING  DATE.
             6. PERFORMING ORGANIZATION CODE
7.AUTHOR(s)  y. 0. Tiernan,  M.  L.  Taylor, S. D. Erk,
  J. G. Solch, G. Van  Ness,  and  J.  Dryden
             8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
                                                           10. PROGRAM ELEMENT NO.
   The Brehm Laboratories and Department of Chemistry
   Wright State University, Dayton,  Ohio  45435
                                                               1BB610
             11. CONTRACT/GRANT NO.

              Contract No. 68-03-2659
12. SPONSORING AGENCY NAME AND ADDRESS
 Industrial  Environmental Research Laboratory
 Office  of  Research and Development
 U.S.  Environmental Protection Agency
 Cincinnati,  OH 45268
                                                           13. TYPE OF REPORT AND PERIOD COVERED
              Final. 10/78 to  3/79
             14. SPONSORING AGENCY CODE
                 EPA/600/12
15. SUPPLEMENTARY NOTES

   Volume II  of a three-volume  series  on  dioxins
16. ABSTRACT
  The  overall  objective of this  research project was to develop  a  unified analytical
  approach  for use in quantifying  ppt levels of tetrachlorodibenzo-p-dioxins (TCDD's)
  in various chemical wastes.  Waste samples from plants manufacturing trichloro-
  phenol,  pentachlorophenol, and hexachlorophene, and from  plants  processing wood
  preservatives were provided  by the EPA.

  The  extraction procedure developed for isolating the TCDD's  from the various types
  of sample matrices is fully  described.  Analysis was accomplished using highly
  specific  and sensitive coupled gas chromatographic-mass spectrometric (GC-MS)
  methods.   Both low and high  resolution MS  techniques were employed.  This method-
  ology is  also described in detail.  The procedures presented in  this report were
  acceptable for most of the industrial  process samples provided.   TCDD's were detected
  and  quantitatively determined  in several of the samples at levels in the ppt to ppm
  range.   One sample, identified as a trichlorophenol stillbottom, was found to con-
  tain 40  ppm TCDD's.  This method was not applicable for wood or  woodlike products
  and  difficulties were also encountered with some samples  that were susceptible
  to emulsion formation in the preparation stages.
17.
                                KEY WORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
                                              b.IDENTIFIERS/OPEN ENDED TERMS
                             COSATI l-ield/Group
  Organic chemicals
  Pesticides
  Chemical analysis
  Industrial wastes
Dioxins;  2,3,7,8-TCDD
Analytical  chemistry
Hazardous waste disposal
07C
07D
13B
18. DISTRIBUTION STATEMENT


    RELEASE TO PUBLIC
19. SECURITY CLASS (ThisReport)
  Unclassified
                                                                         21. NO. OF PAGES
  79
20. SECURITY CLASS (This page I
  Unclassified
                           22. PRICE
EPA Form 2220-1 (Rev. 4-77)   PREVIOUS EDITION is OBSOLETE
             *U.S. GOVERNMENT PRINTING OFFICE: 1981--657-165/0008
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