K 13018
            United States Environmental Protection Agency
            Office of Water Regulations and Standards
            Industrial Technology Division	
            Office of Water
April 1990
Method  1613: Tetra- through
Octa- Chlorinated Dioxins and
Furans by Isotope Dilution
HRGC/HRMS
                         Revision A

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   Introduction
  Method   1613   was  developed  by  the   Industrial   Technology
  Division   (ITD)   within   the  United   States   Environmental
  Protection  Agency's  
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Method  1613    Revision A       October  1990
Tetra- through Octa- Chlorinated  Dioxins  and Furans
by Isotope Dilution  HRGC/HRMS
       1    SCOPE AND APPLICATION

     1.1    Thisjnethod is designed to meet  the  survey
            requirements of the USEPA ITD.   The  method
            is  used to determine  the tetra- through
            octa-  chlorinated  dibenzo-p-dioxins  and
            dibenzofurans   associated  with  the  Clean
            Water Act (as  amended  1987); the Resource
            Conservation and Recovery Act (as amended
            1986); and the Comprehensive  Environmental
            Response, Compensation  and Liability  Act
            (as  amended  1986);  and  other  dioxin  and
            furan compounds  amenable  to  high resolu-
            tion  capillary  column  gas chromatography
            (HRGO/high  resolution  mass  spectrometry
            (HRHS).   Specificity is  provided  for  de-
            termination  of the  17  2,3,7,8-substituted
           polychlorinated  dibenzo-p-dioxins  (PCDD)
           and polychlorinated  dibenzofurans (PCDF).

     1.2    The method  is  based  on  EPA, industry, com-
           mercial   laboratory,  and  academic  methods
           (References  1-6).

     1.3    The compounds  listed 'in  Table  1  may be
           determined  in  waters, soils,  sludges,  and
           other matrices by this method.

     1.4    The- detection  limits  of  the method  are
           usually  dependent  on the  level  of  inter-
           ferences  rather than instrumental  limita-
           tions.   The levels  in  Table  2 typify  the
           minimum  quantities that  can be determined
           in  environmental samples using the method.

     1.5    The  GCHS portions of the method are  for
           use  only by  analysts   experienced  with
           HRGC/HRMS or  under  the  close supervision
           of  such  qualified persons.   Each labora-
           tory  that uses  this method  must demon-
           strate the ability to generate acceptable
           results  using   the   procedure  in Section
           8.2.

      2    SUMMARY OF METHOD

    2.1    Stable isotopically  labeled analogs  of  15
           of the PCDOs and  PCDFs  are added to each
           sample prior to extraction.  Samples con-
           taining  coarse  solids  are prepared  for •
          extraction by grinding or  homogenization.
          Water  samples  are   filtered and   then
          extracted with  methylene  chloride  using
          separatory funnel  procedures; the particu-
           lates from the  water samples,  soils, and
         'other finely  divided  solids are extracted
   2.2
   2.3
   2.4
  2.5
2.5.1
          using  a  combined Soxhlet extraction/Dean-
          Stark  azeotropic distillation  (Reference
          7).   Prior  to  cleanup and analysis, the
          extracts  of the  filtered water  and the
          particulates are combined.
                            37,
                              Gl4-labeled  2,3,7,8-
  After  extraction,
  TCOD  is  added to each extract  to  measure
  the  efficiency  of   the  cleanup process.
  Samples  cleanup  may include back  extrac-
  tion with acid and/or base,  and  gel perme-
  ation, alumina,  silica gel, and  activated
  carbon chromatography.   High performance
  liquid chromatography  (HPLC)  can be used
  for  further  isolation  of   the 2,3,7,8-
  isomers  or   other   specific  isomers  or
  congeners.

  After cleanup, the extract  is concentrated
  to  near  dryness.    Immediately  prior  to
  injection,   two  internal  standards   are
  added to each extract,  and a 1 uL aliquot
  of the extract  is  injected  into  the gas
  chromatograph. The  analytes are separated
 by the GC and detected by  a high reso'lu-
  tion  (>10,000)  mass  spectrometer.    Two
 exact  masses  (m/z's)  are  monitored  for
 each  analyte.    The  isotopically  labeled
 compounds  serve  to  correct   for   the
 variability of the analytical technique.

 Dioxins   and   furans  are  identified  by
 comparing GC   retention times and the ion
 abundance ratios   of the m/z's  with  the
 corresponding  retention  time  ranges  of
 authentic standards   and  the  theoretical
 ion  abundance  ratios of  the exact  m/z's.
 Isomers and congeners are identified  when
 the  retention times  and  m/z   abundance
 ratios  agree  within pre-defined  limits.
 By using  a GC  column or .cotumns capable of
 resolving the  2,3,7,8-substituted isomers
 from   all  other   tetra-  isomers,   the
 2,3,7,8-substituted isomers are identified
 when the  retention time and m/z  abundance
 ratios agree within  pre-defined  limits of
 the retention  times  and exact  m/z ratios
 of authentic standards.

 Quantitative analysis is performed by GCMS
 using'selected ion current profile (SICP)
 areas,  in  one of two  ways.

 For the 15 2,3,7,8-substituted isomers  for
which  labeled  analogs  are available  (see

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          Table  1),  the  GCHS system  is calibrated
          and  the  conpound concentration  is deter-
          mined using an isotope dilution-technique.
          Although   a  labeled  analog  of the  octa-
          chlorinated dibenzofuran  (OCDF)  is avail-
          able, using high resolution mass spectrom-
          etry ft produces an m/z that may interfere
          with the  identification  and quantisation
          of  the unlabeled  octachlorinated  dibenzo-
          p-dioxin  (OCDD).  Therefore,  this  labeled
          analog  has not been  included in the  cali-
          bration standards, and the  unlabeled  OCDF
          is  quantitated  against  the  labeled  OCDD.
          Because the  labeled analog of 1,2,3,7,8,9-
          HxCDD  is  used  as  an  internal  standard
          (i.e.,  not added before extraction of  the
          sample),  it cannot  be  used  to  quantitate
          the  unlabeled compound  by strict  isotope
          dilution procedures.  Therefore, the unla-
          beled  1,2,3,7,8,9-HxCDD   is   quantitated
          using the  average of the  responses of  the
          labeled analogs  of the other  two 2,3,7,8-
          substituted HxCDD's,  1,2,3,4,7,8-HxCDD  and
          1,2,3,6,7,8-HxCDO.   As  a  result, the con-
          centration  of  ,the unlabeled  1,2,3,7,8,9-
          HxCDD   is  corrected  for   the   average
          recovery of the other two  HxCDD's.

 2.5.2    For   non-2,3,7,8-substituted   isomers   and
          the  total  concentrations  of  all   isotners
         within  a  level   of  chlorination  (i.e.,
         total TCDO), concentrations are determined
         using response factors from  the calibra-
         tion  of labeled  analogs  at the same level
         of chlorination.

   2.6   The  quality of  the  analysis  is  assured
         through reproducible calibration and test-
         ing of  the extraction,  cleanup,  and  GCHS
         systems.

    3   CONTAMINATION  AND INTERFERENCES

   3.1    Solvents,   reagents,  glassware,  and other
         sample processing hardware may yield arti-
         facts and/or   elevated  baselines  causing
         misinterpretation  of  chromatograms  (Ref-
         erences  8-9).      Specific  selection   of
         reagents and purification  of  solvents  by
         distillation in  all-glass  systems  may  be
         required.    Where  possible,   reagents  are
         cleaned  by extraction or solvent rinse.

  3.2    Proper cleaning of glassware  is extremely
         important  because  glassware  may not  only
         contaminate the   samples,   but  may  also
         remove  the   analytes  of   interest   by
         adsorption  on the glass surface.

3.2.1    Glassware  should  be  rinsed  with  solvent .
         and  washed with  a  detergent  solution  as
         soon after  use  as is  practical.   Sonica-
         tion of  glassware containing  a detergent
         solution for approximately  30  seconds may
         aid in cleaning.  Glassware with removable
         parts,   particularly   separatory  funnels
         with    teflon    stopcocks,    must    be
         disassembled prior to detergent washing.

3.2.2    After detergent washing,  glassware should
         be immediately rinsed first with methanol,
         then with hot  tap water..   The  tap  water
         rinse  is   followed  by  another  methanol
         rinse,  then  acetone,  and  then  methyl'ene
         chloride.

3.2.3    Do not  bake reusable glassware  in  an  oven
         as a routine part  of  cleaning.   Baking may
         be  warranted  after   particularly dirty
         samples  are  encountered,  but  should  be
         minimized, as  repeated  baking  of glassware
         may  cause   active  sites   on   the glass
         surface  that  will  irreversibly  adsorb
         PCDDs/PCDFs.

3.2.4    Immediately  prior  to  use, Soxhlet  extrac-
         tion  glassware  should  be  pre-extracted
         with  toluene  for  approximately  3 hours.
         See  Section  11.1.2.3.   Separatory  funnels
         should    be    shaken    with    methylene
         chloride/toluene   (80/20   mixture)  for  2
        minutes,  drained,   and  then  shaken   with
        pure methylene chloride for  2 minutes.

  3.3   All  materials  used in  the analysis shall
        be demonstrated  to be free  from interfer-
        ences by  running  reference  matrix blanks
         initially  and  with   each   samole   set
        (samples started  through  the  extraction
        process on  a  given  12-hour shift, to  a
        maximum of  20  samples).    The  reference
        matrix blank' must  simulate,  as  closely as
        possible,  the  sample  matrix unaer test.
        Reagent  water  (Section  6.6.1)   is  used to
        simulate water   samples;   playground  sand
        (Section  6.6.2)   or  white   quartz  sand
       .(Section 6.3.2)  can be used to  simulate
        soils;  filter  paper  (Section  6.6.3)   is
        used  to  simulate  papers  and   similar
        materials; other  materials (Section 6.6.4)
        can be used to simulate  other matrices.

 3.4    Interferences  coextracted '  from   samples
        will  vary  considerably  from  source   to
        source,  depending on the  diversity  of  the
        site being sampled.  Interfering compounds
        may be  present  at  concentrations  several
        orders of  magnitude higher than  the PCDDs
        and PCDFs.    The most   frequently  encoun-
        tered   interferences    are   chlorinated-
        biphenyls,   methoxy  biphenyls,   hydroxy-
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   3.5
  4.1
4.1.1
4.1.2
  4.2
  diphenyl   ethers,   benzylphenyl   ethers,
  potynuctear   aromatics,  and   pesticides.
  Because very  low levels of  PCDDs and PCDFs
  are  measured  by this method,  the elimina-
  tion of  interferences  is  essential.   The
  cleanup steps given  in Section 12  can  be
  used to  reduce  or  eliminate  these  inter-
  ferences   and   thereby  permit   reliable
  determination  of the  PCODs and PCDFs  at
  the  levels shown  in Table 2.

  Each piece of  reusable  glassware should  be
  numbered   in   such   a  fashion   that   the
 ilaboratory  can   associate  all   reusable
  glassware with  the  processing  of a  parti-
  cular  sample.     This  will  assist  the
  laboratory in:   1)  tracking down possible
  sources of contamination   for  individual
  samples,  2)  identifying  glassware  assoc-
  iated  with  highly  contaminated  samples
 'that may  require  extra cleaning,  and 3)
 determining   when  glassware   should  be
 discarded.

 SAFETY

 The  toxicity  or  carcinogenicity of each
 compound  or  reagent used  in  this  method
 has   not    been   precisely   determined;
 however,  each  chemical  compound should be
 treated as a  potential  health  hazard.
 Exposure   to  these  compounds   Should  be
•reduced to  the lowest  possible  level.

 The 2,3,7,8-TCDD isomer has  been found to
 be acnegenic,  carcinogenic,  and  terato-
 genic in laboratory animal  studies.   It is
 soluble in  water to approximately 200 ppt
 and in organic  solvents to  0.14 percent.
 On   the    basis   of    the    available
 toxicological  and  physical  properties  of
 2,3,7,8-TCDD,  all of  the PCDDs  and  PCDFs
 should  be  handled only by highly  trained
 personnel    thoroughly    familiar    with
 handling  and  cautionary  procedures,  and
 who understand the associated risks.

 It  is recommended  that  the  laboratory pur-
 chase dilute  standard  solutions  of  the
 analytes  in  this  method.    However,   if
 primary solutions are prepared,  they  shall
 be  prepared in  a hood,  and  a  NIOSH/MESA
 approved  toxic  gas  respirator  shall-be
 worn when high concentrations are handled.

 The  laboratory  is  responsible  for   main-
 taining a  current awareness  file of OSHA
 regulations regarding the safe handling of
the chemicals specified  in this method.  A
reference   file  of data handling  sheets '
          should  also  be  made  available  to  all
          perspnnel   involved   in   these  analyses.
          Additional    information   on   laboratory
          safety can be  found  in  References  10-13.
          The  references  and bibliography at the end
          of  Reference 13  are  particularly compre-
          hensive   in  dealing   with  the   general
          subject  of  laboratory  safety.

   4.3    The  PCDDs  and PCDFs and  samples  suspected
          to  contain  these  compounds  are  handled
          using  essentially  the  same   techniques
          employed   in   handling   radioactive   or
          infectious  materials.     Well-ventilated,
          controlled    access    laboratories    are
          required.   Assistance in  evaluating  the
          health  hazards  of  particular   laboratory
         conditions  may be obtained  from certain
         consulting  laboratories  and   from   State
         Departments  of  Health  or Labor,  many  of
         which  have  an  industrial  health  service.
         The PCDDs and PCDFs are extremely  toxic  to
          laboratory animals.   Each laboratory must
         develop  a   strict  safety   program  for
         handling the PCDDs and PCDFs.  The follow-
         ing practices are recommended  (References
         2 and 14).

4.3.1    Facility --  When  finely divided  samples
         (dusts, soils, dry chemicals) are handled,
         all   operations   (including  removal   of
         samples from  sample  containers, weighing,
         transferring,   and   mixing),   should  be
         performed in  a  glove box  demonstrated  to
         be leak  tighf or  in  a  fume hood  demon-
         strated to  have adequate air  flow.   Gross
         losses   to   the   laboratory   ventilation
         system  must  not be allowed.   Handling  of
         the   dilute   solutions   normally  used  in
         analytical  and  animal  work  presents  no
         inhalation  hazards except  in the case  of
         an accident.

4.3.2    Protective equipment  -- Throwaway  plastic
         gloves, apron  or tab coat, safety  glasses
         or  mask,  and  a  glove  box  or  fume  hood
         adequate  for  radioactive  work  should  be
         utilized.    During  analytical   operations
         which may give rise to aerosols or dusts,
         personnel should wear respirators equipped
         with  activated   carbon   filters.     Eye
        protection equipment (preferably full  face
        shields) must  be worn  while  working  with
        exposed   samples   or   pure   analytical
        standards.  Latex gloves are commonly used
        to reduce exposure  of   the  hands.   When
        handling  samples  suspected  or  known  to
        contain high  concentrations of  the PCDDs
      •  or PCOFs,  an additional  set  of gloves can '
        also be  worn  beneath the latex gloves.

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   4.3.3    Training -- Workers roust be trained in the
            proper  method  of  removing  contaminated
            gloves and clothing without contacting the
            exterior surfaces.

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

   4.3.5    Confinement --  Isolated  work area,  posted
            with   signs,   segregated   glassware   and
            tools,  plastic  absorbent  paper  on  bench
            tops.

   4.3.6    Effluent vapors  -- The effluents of sample
            splitters  for  the  gas  chromatograph  and
            roughing  pumps  on  the GC/HS should  pass
            through either a column of activated char-
            coal or be bubbled through a trap contain-
            ing oil or high-boiling alcohols.

   4.3.7    Waste Handling and Disposal

 4.3.7.1    Handling   --    Good   technique   includes
            minimizing  contaminated  waste.    Plastic
            bag liners  should be used  in waste cans.
            Janitors  and  other   personnel   must  be
            trained in the safe handling of  waste.

 4.3.7.2    Disposal

4.3.7.2.1    The PCDDs  and PCDFs  decompose  above 800
            °C.    Low-level  waste such as  absorbent
            paper,   tissues,    animal    remains,   and
            plastic gloves may  be  burned in  an appro-
            priate  incinerator.     Gross  quantities
            (milligrams) should be  packaged  securely
            and disposed through commercial  or govern-
            mental  channels  which  are • capable  of
            handling extremely toxic  wastes.

£.3.7.2.2   Liquid  or soluble  waste  should  be  dis-
            solved   in  methanol    or   ethanol   and
            irradiated with  ultraviolet light  with  a
            wavelength greater than 290 nm for several
            days.  (Use f £0  BL lamps  or equivalent.)
            Analyze liquid  wastes  and  dispose  of the
            solutions when the  PCODs and PCDFs  can no
            longer be detected.

   4.3.8    Decontamination

 4.3.8.1    Personal decontamination  -- Use  any  mild
            soap with plenty of scrubbing action.-

 4.3.8.2    Glassware,   tools,    and    surfaces
            Chlorothene NU  Solvent  (Trademark  of the
            Dow Chemical  Company)  is  the  least toxic
            solvent  shown  to  be  effective.    Satis-
            factory  cleaning  may  be  accomplished  by
          rinsing  with  Chlorothene,   then  washing
          with  any  detergent and  water.    If  glass-'
          ware  is first  rinsed with  solvent,  then
          the dish  water may be disposed of  in  the
          sewer.  Given  the cost of disposal,  it  is
          prudent to minimize-solvent wastes.

  4.3.9   Laundry --  Clothing  known  to be  contami-
          nated should be collected  in  plastic bags.
          Persons who convey  the  bags  and  launder
          the  clothing  should  be  advised  of  the
          hazard  and  trained  in   proper  handling.
          The  clothing  may  be put  into  a  washer
          without contact  if the launderer  knows  of
          the potential  problem.   The washer should
          be run  through a  cycle  before being  used
          again for other clothing.

 4.3.10   Wipe tests  -- A useful method of  determin-
          ing cleanliness of  work  surfaces  and tools
          is  to wipe  the  surface  with  a  piece  of
          filter paper.   Extraction and  analysis  by
          GC can achieve a  limit of detection of  0.1
          ug per  wipe.   Less than 0.1  ug   per  wipe
          indicates acceptable  cleanliness;  anything
          higher  warrants  further cleaning.    More
          than  10 ug  on  a wipe constitutes  an acute
          hazard and  requires prompt cleaning before
          further  use  of   the  equipment   or  work
          space,  and  indicates   that  unacceptable
          work practices have been employed.

 4.3.11   Accidents --  Remove contaminated  clothing,
          immediately,  taking  precautions   not   to
          contaminate  skin  or other articles.   Wash
          exposed  skin  vigorously  and  repeatedly
          until medical attention  is obtained.

      5   APPARATUS AND MATERIALS

    5.1   Sampling   equipment   for    discrete   or
          composite sampling.

  5.1.1   Sample bottles and caps

5.1.1.1   Liquid samples  (waters,  sludges and  simi-
          lar  materials   containing  five   percent
          solids  or   less)  --  Sample  bottle,  amber '
          glass, 1.1 L minimum, with,screw cap.

5.1.1.2   Solid samples  (soils, sediments,   sludges,
          paper  pulps,  filter  cakes,   compost,  and
          similar materials that  contain more  than
          five  percent  solids)  --   Sample  bottle,
          wide mouth, amber glass,  500" mL minimum.

5.1.1.3   If   amber   bottles   are  not  available,
          samples shall be protected from light.

5.1.1.4   Bottle  caps  --  Threaded  to  fit  sample
          bottles.  Caps shall be  lined with  Teflon.
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  5.1.1.5

 5.1.1.5.1


 5.1.1.5.2
    5.1.2
     5.2

   5.2.1

     5.3

   5.3.1



   5.3.2

   5.3.3
  5.3.4


  5.3.5

5.3.5.1


5.3.5.2

  5.3.6

5.3.6.1

5.3.6.2
  Cleaning

  Bottles are detergent  water  washed, then
  solvent rinsed before use.

  Liners  ape detergent  water  washed, then
  rinsed with reagent  water (Section  6.6.1)
  and    then   solvent,    and    baked   at
  approximately 200 °C  for one  hour minimum,
  prior to-use.

  Compositing  equipment   --  Automatic - or
  manual-  compositing  system  incorporating
  glass   containers   cleaned   per   bottle
  cleaning procedure above.  Glass or Teflon
  tubing only shall be used.  If the sampler
  uses a peristaltic pump,  a minimum length
  of  compressible silicone rubber tubing may
  be  used in the pump only.  Before use, the
  tubing shall  be  thoroughly  rinsed  with
  methanol,   followed  by  repeated  rinsings
  with  reagent   water   to  minimize  sample
  contamination.   An integrating  flow  meter
  is  used to collect proportional  composite
  samples.

  Equipment  for glassware cleaning

,  Laboratory sink with overhead fume hood

  Equipment  for sample preparation

  Laboratory  fume hood of sufficient size to
  contain  the sample  preparation equipment
  listed below

 Glove box (optional)

 Tissue  homogenizer  --  VirTis  Model  45
 Macro  homogenizer   (American   Scientific
 Products   H-3515,   or   equivalent)   with
 stainless  steel  Macro-shaft  and  Turbo-
:shear blade.
             »
 Meat  grinder  --  Hobart,  or  equivalent,
 with 3-5 mm holes  in  inner plate.

 Equipment for determining  percent moisture

 Oven, capable of maintaining a  temperature
 of 110 ±5 °C.

 Oessicator

 Balances

Analytical  -- Capable of weighing 0.1 mg.

Top  loading  — Capable of weighing 10 mg.
    5.4    Extraction apparatus

  5.4.1    Water samples
5.4.1.1    pH    meter,
           electrode.
with    combination   glass
5.4.1.2   pH  paper,  wide  range  (Hydrion Papers, -ar;
          fquivalent).

5.4.1.3   Graduated cylinder, 1 L capacity

5.4.1.4   1 L  filtration  flasks with  side arm,  for
          use in vacuum filtration of water samples.

5.4.1.5   Separatory funnels  --  250,  500,  and 2000
          ml, with Teflon stop cocks.
                                                             5.4.2    Soxhlet/Dean-Stark     . (SOS)
                                                                      (Figure  1)
                                           extractor
                                                                   FIGURE 1   Soxhlet/Dean-Stark Extractor

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  5.4.2.1    Soxhtet  --  50  mm  i.d.f  300 raL  capacity
            with 500  mL  flask  (Cat-Glass  LG-6900,  or
            equivalent,  except substitute 500 ml. round
            bottom  flask  for  300   ul  flat  bottom
            flask).

5.4.2.2     Thimble -- A3  x 123 to  fit Soxhlet (Cal-
            Glass LG-6901-122,  or  equivalent).

 5.4.2.3    Moisture trap -- Dean  Stark or Barret with
            Teflon stopcock,  to fit Soxhlet.

 5.4.2.4    Heating mantle  --  Hemispherical,  to  fit
            500  mL  round  bottom flask  CCal-Glass  LG-
            8801-112,  or  equivalent).

 5.4.2.5    Variable  transformer   --   Powerstat   (or
            equivalent),  110  volt,  10  amp.

   5.4.3    Beakers, 400-500  mL

   5.4.4    Spatulas -- Stainless steel

     5.5    Filtration apparatus

   5.5.1   Pyrex  glass  wool -- Solvent extracted by
           SOS  for  three   hours  minimum.     (NOTE:'
           Baking  glass  wool may  cause active sites
           that      will      irreversibly      adsorb
           PCOOs/PCOFs.)

   5.5.2   Glass funnel — 125-250 mL

   5.5.3   Glass fiber filter paper (Whatman GF/D, or
           equivalent)

   5.5.4   Drying  column  -- 15 to 20  irm  i.d.  Pyrex
           chromatographic   column   equipped   with
           coarse glass frit or glass wool plug.

   5.5.5   Buchner funnel, 15 cm.

   5.5.6   Glass fiber filter paper for above.

   5.5.7   Pressure filtration apparatus -- Millipore
           YT30 142 HU,  or equivalent.
             »
    5.6   Centrifuge apparatus

   5.6.1    Centrifuge --  Capable  of   rotating 500  mL
           centrifuge bottles   or  15  mL  centrifuge
           tubes at 5,000 rpn minimum

   5.6.2   Centrifuge bottles  —  500 mL, with  screw
           caps,  to fit centrifuge

   5.6.3    Centrifuge tubes  --  12-15 mL, with  screw
           caps,  to fit centrifuge
     5.7   Cleanup apparatus

   5.7.1   Automated   gel   permeation   chromatograph
           (Analytical    Biochemical    Labs,    Inc,
           Columbia, MO, Model  GPC Autoprep  1002,  or
           equivalent).

 5.7.1.1   Column -- 600-700  mm x 25 mm  i.d.,  packed
           with  70  g  of  SX-3   Bio-beads   (Bio-Rad
           Laboratories,     Richmond,     i;A,      or
           equivalent).

 5.7.1.2   Syringe, 10 mL,  with Luer fitting.

 5.7.1.3   Syringe  filter   holder,  stainless  steel,
           and glass fiber  or Teflon filters (Gelman
           4310, or equivalent).

 5.7.1.4   UV  detectors --   254-nm,  preparative   or
           semi-prep flow cell:  (Isco,  Inc.,  Type  6;
           Schmadzu,  5  mm path  length;  .Beckman-Altex
           152W, 8  uL  micro-prep  flow  cell,  2   tm
           path; Pharmacia UV-1, 3 mm flow cell;  LDC
           Milton-Roy   UV-3,    monitor   #1203;    or
           equivalent).

   5.7.2   Reverse   phase  high   performance   liquid
           chromatograph

 5.7.2.1    Column oven  and  detector  --  Perkin-Elmer
           Model LC-65T  (or  equivalent)  operated  at
           0.02 AUFS  at  235  nm.

 5.7.2.2   Injector  --  Rheodyne 7120  (or  equivalent)
           with 50 uL sample  loop.

 5.7.2.3    Column --  Two 6.2  x   250  run  Zorbax-ODS
           columns   in   series   (DuPont   Instruments
           Division,  Wilmington, DE, or  equivalent),
           operated at 50 °C with  2.0 mL/min  methanol
           isocratic effluent.

 5.7.2.4    Pump -- Altex 110A  (or equivalent).

  5.7.3    Pipets

5.7.3.1    Disposable,  Pasteur,  1SO  mm  x  5  mm i.d.
           (Fisher    Scientific    13-678-6A,    or
           equivalent).

5.7.3.2    Disposable,   serological,   10  mL   (6  mm
           i.d.).

  5.7.4    Chromatographic columns

5.7.4.1    150  irni x  8 mm i.d.,  (Kontes  K-420155,  or
           equivalent)  with  coars;e~ glass  frit  or
           glass  wool plug and 250 mL reservoir.
.[
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  5.7.4.2    200 mm  x 15  mm i.d.,  with coarse  glass
             frit   or  glass  wool  plug and  250  mt
             reservoir.

    5.7.5    Oven  --  For  storage of adsorbents, capable
             of  maintaining a temperature of  130 ±5 °C.
      5.8   Concentration  apparatus
    5.8.1
  5.8.1.1
  5.8.1.2
            Rotary  _evaporator
            American   Scientific
            equivalent,   equipped
            temperature water bath.
                            Buchi/Brinkman-
                        No.   E5045-10   or
                        with   a   variable
A vacuum source is required for use  of  the
rotary  evaporator.    It must  be  equipped
with  a  shutoff  valve  at  the evaporator,
and preferably, have a vacuum gauge.

A reci.rculating water pump and chiller  are
recommended,  as  use  of  tap  water   for
cooling   the   evaporator   wastes  large
volumes   of  water   and   can   lead    to
inconsistent    performance    as    water
temperatures and pressures vary.
  5.8.1.3   Round bottom  flasks -- 100  mL  and 500 mL
            or   larger,   with   ground  glass   fitting
            compatible with the rotary evaporator.

    5.8.2   Kuderna-Oanish (K-D)

  5.8.2.1   Concentrator tube--10mL, graduated  (Kontes
            K-570050-1025,    or    equivalent)    with
            calibration   verified.      Ground   glass
            stopper  (size  19/22  joint)  is  used  to
            prevent evaporation of extracts.

  5.8.2.2   Evaporation   flask--500  mL  {Kontes   K-
            570001-0500,  or  equivalent), attached  to
            concentrator tube with  springs  (Kontes  K-
            662750-0012).

  5.8.2.3   Snyder column--three ball  macro (Kontes  K-
            503000-0232, or equivalent)."

  5.8.2.4   Boiling chips

5.8.2.4.1   Glass  or  silicon  carbide--approx  10/40
            mesh, extracted  with  methylene  chloride
            and baked at 450  °C  for one h minimum.
5.8.2.4.2
 5.8.2.5
            Teflon     (optional)--extracted
            methylene chloride.
                                      with
           Water  bath—heated,  with 'concentric  ring
           cover,    capable    of   • maintaining    a
           temperature  within +/- 2 "C,  installed  in
           a  fume hood.
 5.8.3    Nitrogen blowdown  apparatus  --  Equipped
          with  water  bath controlled at 35-40 °C (N-
          Evap,  Organomation  Associates, Inc., South
          Berlin,  HA,  or equivalent), installed in a
          fume  hood.

 5.8.4    Sample vials -- Amber glass, 2-5  mL  with
          Teflon-lined screw  cap.

   5.9    Gas chromatograph  --  Shall  have  splitless
          or on-column injection port  for  capillary
          column,     temperature     program     with
          isothermal hold, and  shall  meet all  of the
          performance  specifications  in  Section  7.

 5.9.1    GC  Column  for  PCDDs and  PCDFs  and  for
          isomer specificity  for 2,3,7,8-TCDD  --   60
          ±5 m   x  0.32  ±0.02 ran  i.d.;  0.25  urn  5%
          phenyl,  94% methyl,  1%  vinyl   silicone
          bonded phase fused  silica capillary  column
          (J & U DB-5, or equivalent).

 5.9.2    GC  Column   for  isomer • specificity   for
          2,3,7,3-TCDF —  30 ±5 m x  0.32  ±0.02 ran
          i.d.;   0.25  urn  bonded phase  fused  silica
          capillary  column   (J  &   W  DB-225,   or
          equivalent).

 5.10   Mass   spectrometer   --'28-40  eV  electron-
          impact  ionization,   shall " be  capable  of
          repetitively  selectively  monitoring   12
         exact   m/z's minimum  at  high  resolution
         (>10,000) during a period of approximately
         1  second,   and  shall  meet  all   of  the
         performance  specifications in Section 7.

 5.11    GCHS   interface  --   The mass  spectrometer
         (MS)  shall  be  interfaced  to the  GC such
         that   the end   of   the  capillary  column
         terminates  within 1  cm of  the  ion source
         but does  not intercept the electron or  ion
         beams-.

 5.12    Data   system --  Capable  of  collecting,
         recording and storing  MS data.

    6    REAGENTS  AND  STANDARDS

  6.1    pH adjustment and back extraction

6.1.1    Potassium hydroxide  --   Dissolve   20   g
         reagent grade KOH in 100 mL  reagent water.

6.1.2    Sulfuric  acid -- Reagent  grade (specific
         gravity 1.84).

6.1.3    Sodium  chloride  --  Reagent grade,  prepare
         a five percent  (w/v)  solution in reagent
         water.

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     6.2   Solution drying and evaporation

   6.2.1   Solution drying — Sodium sulfate, reagent
           grade, granular anhydrous  (Baker 3375, or
           equivalent),    rinsed   with    methylene
           chloride- (20  "mL/g),  baked  at 400  "C for
           one hour minimum,  cooled in a dessicator,
           and stored  in a pre-cleaned  glass bottle
           with screw cap that prevents moisture from
           entering.   If  after  heating the  sodium
           sulfate develops a noticeable grayish cast
           (due  to  the  presence  of  carbon in  the
           crystal matrix), that  batch of  reagent is
           not  suitable   for  use  and  should  be
           discarded.    Extraction  with   methylene
           chloride  (as  opposed  to simple rinsing)
           and baking  at  a   lower temperature  may
           produce sodium  sulfate  that is suitable
           for use.

  6.2.2   Prepurified  nitrogen

    6.3   Extraction

  6.3.1    Solvents  --  Acetone, toluene, cyclohexane,
           hexane,    nonane,    methanol,    methylene
           chloride,  and nonane:  distilled-in-glass,
           pesticide quality,  lot  certified  to  be
           free of interferences.

  6.3.2   White   quartz  sand,   60/70   mesh •--   For
           Soxhlet/Oean-Stark   extraction,   (Aldrich
           Chemical   Co,  Milwaukee     WI     Cat   No.
           27,437-9,  or equivalent).   Bake  at 450 "C
           for four  hours minimum.

    6.4    GPC  calibration   solution   --   Solution
           containing  3QO  mg/mL  corn  oil,  15 mg/ml
           bis(2-ethylhexyl)   phthalate,  1.4  mg/ml
           pentachlorophenol,  0.1  mg/mL perylene,  and
           0.5 mg/mL sulfur

    6.5    Adsorbents for sample  cleanup

  6.5.1    Silica gel

6.5.1.1    Activated silica gel -- Bio-Si I A,  100-200
           mesh  (Bio-Rad  131-1340,   or  equivalent),
           rinsed  with  methylene  chloride,  baked  at
           180 "C for one  hour minimum,  cooled in-a
           dessicator,  and stored  in  a pre-cleaned
           glass  bottle with  screw  cap that prevents
           moisture from  entering.

6.5.1.2    Acid   silica   gel   (30   percent  w/w)   --
           Thoroughly  mix  44.0   g  of  concentrated
           sulfuric  acid' with  100.0 g of  activated
           silica gel in  a clean  container.  Break up
           aggregates  with  a  stirring rod  until a
           uniform mixture  is  obtained.   Store in a
           screw-capped bottle with Teflon-lined cap.

 6.5.1.3    Basic silica gel -- Thoroughly mix 30 g of
           1N  sodium   hydroxide   with  100   g  of
           activated silica gel in a clean container.
           Break up  aggregates with  a  stirring rod
           until   a  uniform   mixture  is  obtained.
           Store  in   a  screw-capped   bottle  with
           Teflon-lined cap.

   6.5.2    Alumina   —  Either  one  of  two types  of
           alumna,  acid or basic, may be used in the
           cleanup  of sample extracts,  provided that
           the laboratory  can  meet  the  performance
           specifications for the recovery of labeled
           compounds described  in Section 8.3.   The
           same type of alumina must  be used  for all
           samples,    including .  those   used    to
           demonstrate  initial  precision and accuracy
           (Section  8.2)  and  ongoing  precision  and
           accuracy  (Section 14.5).

 6.5.2.1    Acid .alumina --  Bio-Rad  Laboratories  132-
           1340 Acid Alumina AG  4 (or equivalent).
           Activate  by  heating  to  130  °C for 12 hours
           minimum.

 6.5.2.2    Basic  alumina  --  Bio-Rad Laboratories 132-
           1240 Basic Alumina AG  10 (or equivalent).
           Activate  by heating  to  600  °C for  24 hours
           minimum.      Alternatively,  activate   by
           heating alumina  in a tuibe  furnace  at  650-
           700 "C under an  air  flow of  approximately
           400 cc/min.   Do  not  heat over 700  °C,  as
           this can  lead  to  reduced  capacity  for
           retaining  the analytes.  Store at  130  °C
           in a covered flask.   Use within  five  days
           of baking.

  6.5.3    AX-21/Celite

6.5.3.1    Activated   carbon   —   AX-21    (-Anderson
           Development   Company,   Adrian,   MI,    or
           equivalent).   Prewash  with  methanol  and
           dry in vacuo at 110  "C.
6.5.3.2
6.5.3.5
Celite   545
equivalent).
(Supelco   2-0199,    or
Thoroughly  mix 5.35  g AX-21  ,and  62.0 g
Celite 545  to  produce a 7.9% w/w mixture.
Activate  the mixture  at   130  °C  for  six
hours minimum.  Store in a dessicator.
    6.6   Reference matrices

  6.6.1   Reagent water  --  Water in which the PCDDs
          and  PCDFs and interfering  compounds  are
          not detected by this method.
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 6.6.2   High solids reference matrix  -- Playground
         sand  or  similar  material   in which   the
         PCDDs and  PCDFs  and interfering compounds
         are not  detected by this method.   May be
         prepared  by   extraction  with  methylene
         chloride and/or  baking  at  450 °C for four
         hours minimum.

 6.6.3   Filter paper  --  Gelman  type A (or equiva-
         lent) glass  fiber  filter  paper  in  whfch
         the PCDDs  and PCDFs and interfering com-
         pounds  are  not  detected by  this  methoa.
         Cut the paper to simulate the surface area
         of the paper sample being tested.

 6.6.4   Other  matrices  --  This  method  may  be
         verified on  any  matrix  by  performing  the
         tests given in Section  8.2.   Ideally,  the
         matrix  should be  free   of  the PCDDs   and
         PCDFs, but in no case shall the background
         level   of  the  PCDOs  and  PCDFs  in   the
         reference matrix exceed  three times   the
         minimum levels given  in Table 2.   If  low
         Background levels  of  the PCODs  and  PCDFs
         are present  in  the reference  matrix,   the
         spike  level   of   the   analytes   used  ;n
       .  Section 8.2 should be increased to provice
         a  spike-to-background  ratio  in  the  range
         of 1/1 to 5/1  (Reference 15).

  6.7   Standard solutions  --  Purchased as  solu-
         tions  or  mixtures  with certification  to
         their  purity,  concentration,  and  authen-
         ticity,   or  prepared  from  materials  of
         known  purity  and  composition.   If. compound
         purity  is  98  percent   or  greater,   the
         weignt may be  used without correction  to
         compute  the concentration of the standara.
         When  not being used, standards  are  stored
       ,  in the- dark at room temperature  in  screw-
       ;  capped vials  with  Teflon-lined caps.    A
         mark  is  placed on the vial  at  the  level  of
         the'solution  so  that solvent  evaporation
         loss can be detected.   If solvent  loss  has
         occurred, the  solution should  be replaced.

  6.8    Stock  solutions
6.8.1
        Preparation  --  Prepare  in  nonane per  the
        steps  below or  purchase as  dilute  solu-
        tions   (Cambridge  Isotope   Laboratories,
        Cambridge,  MA,   or equivalent).   Observe
        the  safety precautions  in  Section 4,  and
        the  recommendation  in Section 4.1.2.
6.8.2   Dissolve  an  appropriate amount of  assayed
        reference  material    in   solvent.      For
        example,  weigh  1-2 mg  of 2,3,7,3-TCDD  to
        •three  significant  figures   in  a   10   ml
        ground  glass  stoppered  volumetric  flask
        and  fill  to the  mark with nonane.   After
        the  TCDD  is  completely dissolved,  transfer
        the  solution to  a clean  15  ml vial  with
        Teflon-lined cap.

6.8.3   Stock standard solutions  should be checked
        for  signs  of  degradation  prior  to  the
        preparation  of  calibration or  performance
        test standards.   Reference standards  that
        can  be  used to determine the  accuracy  of
        calibration  standards  are available  from
        Cambridge Isotope  Laboratories.

  6.9   Secondary  standard  --  Using  stock  solu-
        tions  (Section   6.8),  prepare secondary
        standard  solutions   containing the   com-
        pounds and concentrations  shown in  Table 4
        in nonane.

 6.10   Labeled  compound   stock standard  --   From
        stock  standard    solutions   prepared   as
        above,  or from purchased  mixtures,  prepare
        this standard to  contain  the labeled  com-
        pounds  at  the  concentrations shown  in
        Table  4   in  nonane.    This  solution  is
        diluted with acetone  prior to use  (Section
        10.3.2).

 6.11    Cleanup standard  - Prepare   Cl.-2,3,7,3-
        TCDO at the concentration shown  in  Table 4
        in rionane.

 6.12    Internal  standard  --  Prepare at the   con-
        centration shown  in Table  4 in nonane.

 6.13    Calibration standards (CS1 through CSS) --
        Combine  the  solutions  in  Sections  6.9,
      .  6.10, 6.11, and 6.1,2 to  produce the   five
        calibration solutions shown in  Table  4 in
        nonane.   These solutions  permit the rela-
        tive response (labeled to unlabeled)  and
        response  factor  to be measured  as  a func-
        tion of concentration.   The  CS3 standard
        is  used   for  calibration   verification
        (VER).

6.14    Precision  and recovery  standard (PAR)  --
        Used for  determination of  initial (Section
        8.2)  and  ongoing  (Section  14.5) precision
        and  accuracy.   This solution  contains  the
        analytes and  labeled  compounds at the  con-
        centrations  listed in  Table  4  in  nonane.
        This  solution  is  diluted with  acetone
        prior to use  (Section  10.3.4).

6.15    GC  retention  time window  defining solu-
        tions --  Used to  define the beginning  and
        ending retention times for the  dioxin  and
        furan isomers.

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 6.15.1   DB-5  column window  defining standards  --
          Cambridge  Isotope  Laboratories  ED-1732-A
          (dioxins)   and   ED-1731-A  (furans),   or
          equivalent,   containing    the    compounds
          listed  in Table 5.

   6.16   Isomer  specificity test standards -- Used
          to demonstrate  isomer specificity for  the
          2,3,7,8-tetra-   isomers   of  dio in   and
          furan.

 6.16.1   Standards for the DB-5  column --  Cambridge
          Isotope  Laboratories  ED-908, ED-SJpS-C,  or
          ED-935,  or   equivalent,  containing '  the
          compounds listed  in Table  5.

 6.16.2   Standards   for   the   DB-225   column
          Cambridge  Isotope Laboratories  EF-937  or
          EF-938,  or   equivalent,  containing   the
          compounds listed  in Table  5.

   6.17   Stability   of    solutions  --   Standard
          solutions  used for  quantitative purposes
          (Sections  6.9-6.14)   shall  be  analyzed
          within  48  hours  of  preparation  and  on a
          monthly  basis  thereafter  for   signs   of
          degradation.      Standards  will  remain
          acceptable   if   the  peak  area   at   the
          quantitation   m/z   remains  within    ±15
          percent  of   the   area  obtained  in   the
          initial  analysis  of  the  standard.     Any
          standards  failing to meet this  criterion
          should   be   assayed   against    reference
          standards,  as in Section 6.8.3.,  before
          further use.

      7   CALIBRATION

    7.1   Assemble   the  GCHS  and   establish   the
          operating conditions necessary to meet  the
       •'  relative retention  time specifications  in
          Table 2.

  7.1.1   The following  GC  operating conditions  may
          be  used  for  guidance  and  adjusted   as
          needed to meet the relative  retention time
          specifications in Table 2:
          Injector tenp:  270 °C
          Interface temp:  290 °C
          Initial temp and time: 200 °C,  2 min
          Temp Program:  200-220 °C  at 5 °C/min
                         220 °C for  16 min
                         220-235 °C  at 5 °C/min ,
                         235 °C for  7 min
                         235-330 °C  at 5 °C/min

          NOTE:   All  portions  of the column which
          connect  the GC  to  the ion source  shall
          remain   at   the   interface  temperature
          specified   above   during   analysis,    to
          preclude  condensation   of   less  volatile
          compounds.

  7.1.2   Mass   spectrometer   (MS)   resolution
          Obtain  a  selected   ion current   profile
          (SICP)  of  each analyte  in  Table 4 at  the
          two exact masses  specified in Table 3  and
          at >10,000 resolving  power  by injecting an
          authentic standard of the PCDDs and PCDFs
          either  singly or as  part  of a  mixture in
          which  there  is  no  interference   between
          closely   eluted   components,   using   the
          procedure in  Section  13.

7.1.2.1   The analysis  time for PCDDs and PCDFs  may
          exceed the long-term  mass stability of  the
          mass spectrometer.  Because the  instrument
          is  operated   in  the high-resolution mode,
          mass drifts  of a  few ppm  (e.g., 5 ppm in
          mass) can have serious  adverse  effects on
          instrument  performance.     Therefore,,   a
          mass-drift  correction  is  mandatory.     A
          lock-mass ion from the  reference  compound
          (PFK)   is   used   for   tuning   the  mass
          spectrometer.     The   lock-mass   ion   is
          dependent  on  the  masses   of  the   ions
          monitored within each descriptor,  as shown
        •  in  Table  3.    The level  of the reference
          compound   (PFK)   metered   into   the   ion
          chamber during HRGC/HRMS  analyses should
          be  adjusted  so that  the amplitude of  the
          most intense  selected lock-mass'ion signal
          (regardless of the descriptor numbe--,  does
          not exceed  10  percent   of  the -fuU- scale
          deflection  for a  given  set  of  detector
          parameters.     Under   those  conditions,
          sensitivity   changes   that   might . occur
          during   the    analysis    can   ba   more
          effectively monitored.    NOTE:   Excessive
          PFK (or any other  reference substance)  may
          cause noise  problems  and contamination  of
          the ion source resulting in an increase in
          time lost in  cleaning the source.

7.1.2.2   By  using  a PFK  molecular  leak,  tune  the
          instrument to meet  the  minimum  required
          resolving  power  of  10,000  (10   percent
          valley) at m/z 304.9824  (PFK) or any other
          reference  signal  close  to  m/z  303.9016
          (from TCDF).   By  using  the peak  matching
          unit and  the PFK reference  peak, verify
          that the exact mass  of  m/z 380.:9760 (PFK)
          is within 5 ppm of the required value.

    7.2   Ion  abundance  ratios,   minimum   levels,
          signal-to.-noise   ratios,   and    absolute
          retention   times   --   Inject   the    CS1
.1
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           calibration _solution  (Table  4)  per  the
           procedure in Section 13 and the conditions
           in Table 2.

  7.2.1    Measure  the  SICP areas  for  each  analyte
           and  compute  the   ion  abundance   ratios
           specified  in  Table   3A.     Compare  the
           computed ratio  to. the  theoretical  ratio
           given 'in Table 3A.

7.2.1.1  '  The groups  of m/z's  to  be monitored  are
           shown  in   Table  3.     Each  group   or
           descriptor  shall be monitored  in  succes-
           sion  as  a  function  of GC  retention  time to
           ensure   that  all  PCDDs   and  PCDFs   are
           detected.      The   theoretical   abundance
           ratios  for  the  m/z's  are  given in  Table
           3A, along  with the control  limits  of  each
           ratio.

7.2.1.2    The mass spectrometer shall be operated in
           a  mass  drift  correction  mode, using  per-
           fluorokerosene  (PFK)  to   provide   lock
           masses.   The  lock mass for each group  of
           m/z's is shown in Table 3.   each  lock  mass
           shall be monitored and shall  not vary  by
           more  than   ±10   percent   throughout   its
           respective  retention time window.-  Varia-
           tions of the  lock   mass  by more  than  10
           percent  indicate  the  presence  of coeluting
           interferences   that    may- significantly
         1  reduce   the   sensitivity  of   the  mass
           spectrometer.  .   Re-injection  of  another
          aliquot  of  the   sample  extract   will  not
           resolve  the problem.   Additional cleanup
          of  the extract may  be  required  to remove
          the interferences.

  7.2.2   All PCDDs and  PCDFs  shall be within their
          respective  ratios;   otherwise,  the  mass
          spectrometer  shall  be  adjusted   and  this
         •test  repeated  until  the  m/z  ratios  fall
          within  the  limits  specified.    If  the
          adjustment  alters the  resolution of  the
          mass  spectrometer,   resolution  shall  be
          verified  (Section 7.1)  prior  to  repeat of
          the test.

  7.2.3   Verify that the HRGC/HRMS instrument meets
          the minimum  levels in  Table 2.  The peaks
      •  .representing  both unlabeled   and  labeled
          analytes  in the calibration standards must
          have a signal-to-noise ratio (S/N) greater
          than or   equal  to  10; otherwise,   the  mass
          spectrometer shall  be  adjusted  and  this
          test repeated until   the minimum  levels in
          Table  2 are met.
7.2.4   The  absolute   retention   time   of
                                                                                                                          "12,
                                                                                    1,2,3,4-TCOD (Section  6.12)  shall  exceed
                                                                                    25.0 minutes on  the DB-5 column,  and the
                                                                                    retention time  of   C.J.-1,2,3,4-TCDD shall
                                                                                    exceed 15.0 minutes on  the  08-225 column;
                                                                                    otherwise,   the  GC   temperature  program
                                                                                    shall  be adjusted  and this  test repeated
                                                                                    until  the  above-stated  minimum  retention
                                                                                    time criteria are met.

                                                                             7.3    Retention  time  windows  --  Analyze  the
                                                                                    window defining  mixtures  (Section  6.15)
                                                                                    using  the procedure in  Section 13 (Figures
                                                                                    2A-2D).   Table 5 gives  the  elution  order
                                                                                    (first/last) of the compound pairs.

                                                                             7.4    Isomer specificity

                                                                           7.4.1    Analyze   the   isomer   specificity   test
                                                                                    standards    (Section    6.16)    using    the
                                                                                    procedure in Section 13.

                                                                           7.4.2    Ccmoute the  percent valley between the  GC
                                                                                    peaks,  that  elute  most  closely  to   the
                                                                                    2,3,7,3-  TCDD  and  TCDF  isomers,  on  their
                                                                                    respective columns,"per Figure 3.

                                                                           7.4.3   Verify  that  the   height  of   the  valley
                                                                                   between  the most  closely  eluted  isomers
                                                                                   and  the  2,3,7,8-  isomers is  less than 25
                                                                                   percent (computed as 100  x/y  in Figure 3).
                                                                                 •   If the valley  exceeds  25 percent,' adjust
                                                                                   the  analytical  conditions and  repeat the
                                                                                   test or replace the GC  col-umn and recali-
                                                                                   brate (Section  7.2 through 7.4).

                                                                             7.5   Calibration  with   isotope   dilution
                                                                                   Isotope  dilution   is   used   for   the  15
                                                                                   2,3,7,8-substituted  PCDOs and  PCDFs  with
                                                                                   labeled compounds  added  to  the  samples
                                                                                   prior to extraction, and  for 1,2,3,7,8,9-.
                                                                                   HxCOO and OCDF  (see Section 16.1).   The
                                                                                   reference   compound   for  each   unlabeled
                                                                                   compound is  shown  in Table 6.

                                                                          7.5.1    A  calibration   curve   encompassing   the
                                                                                   concentration range  is  prepared  for  each
                                                                                   compound  to be  determined.    The  relative
                                                                                   response  (RR) (unlabeled  to  labeled)  vs.
                                                                                   concentration  in  standard   solutions   i's
                                                                                   plotted or computed  using  a  linear regres-
                                                                                   sion.    Relative  response  is  determined
                                                                                   according   to  the  procedures  described
                                                                                   below.   A  minimum of five data  points  are
                                                                                   employed for calibration.

                                                                          7.5.2    The  relative response  of  each unlabeled
                                                                                   PCOD/PCDF   and   its   labeled  analog   is
                                                                                   determined  using  the  area   responses  of
                                                                                                                          11

-------
                   "   6-MAY-88    Sir Voltage 705    Sys: DBSUS
           Sample 1 Injection 1    Group 2    Mass 303.9016
       100
       80
       60
       40-
       20-
                  1,3,6,8-TCDF
                                                    1,2,8,9-TCDF
                                                                     Norm:    3044
I
I
I
t
         25:20  26:40  28:00  29:20  30:40   32:00  33:20  34:40  36:00   37:20  38:40
                     6-MAY-88    Sir: Voltage 705    Sys: DB5US
          Sample 1 Injection 1    Group 2    Mass 319.8965
       lOOi
                                                                      Norm: '   481
                                                    1,2.8.9-TCDD
          25:20  26:40  28:00  29:20   30:40- 32:00/33:20  34:40  36:00  37:20   38:40
                FIGURE 2A  First and Last Eluted Tetra- Dioxin and Furan Isomers
12
[

[

[

te».

[

[

[

-------
                6-MAY-88    Sir: Voltage 705    Sys: DB5US
     Sample 1 Injection 1    Group 2    Mass 339.8597
    100
    80
    60
    40
    20
             1,3,4,6,8-PeCDF
                                              Norm:    652
                                               1,2,3,8,9-PeCDF
      29:20  30:40  32:00  33:20  34:40  36:00  37:20  38:40
                 6-MAY-88    Sir: Voltage 705    Sys: DB5US
      Sample 1 Injection 1    Group 2    Mass 355.8546
  100
   80
   60
  40-
  20-
                  1,2,4,7,9-PeCDD-
Norm:    503
                                                1,2,3,8,9-PeCDD
    \
    29:20  30:40  32:00  33:20  34:40  36:00  37:20  38:40
FIGURE 2B  First and Last Eluted Penta- Dioxin and Furan Isomers
                                                                            13

-------
                           6-MAY-88    Sir: Voltage 705    Sys: DB5US
               Sample 1 Injection 1    Group 3    Mass-373.8208
          100
           80
          60
          40-
          20-
Norm::    560
                    1,2,3,4,6,8-HxCDF
                                                               1,2,3,4,8,9-HxCDF
               >	L-
             39:30  40:00 40:30  41:00  41:30  42:00  42:30 43:00  43:30  44:00  44:30
                        6-MAY-88    Sir: Voltage 705   Sys: DB5US
             Sample 1 Injection 1    Group 3    Mass 389.8156
        100
         00
         60
         40
        20-
                            1,2,4,6,7,9/1,2,4,6,8,9-HxCDD
                                                                       Norm:    384
                                                            1.2,3,4,6,7-HxCDD
            39:30  40:00  40:30  41:00  41:30  42:00 42:30  43:00  43:30  44:00  44:30
                 FIGURE 2C  First and Last Eluted Hexa- Dioxin and Furan Isomers
14

-------
              6-MAY-88    Sir: Voltage 705    Sys: DB5US
  Sample 1 Injection 1    Group 4    Mass 407.7818
  100
   80
   60'
   40'
   20-
              1,2.3,4,6,7,8-HpCDF
                                       Norm:    336
1,2,3,4,7,8,9-HpCDF
     45:20   46:40   48:00   49:20   50:40   52:00   53:20   54:40   56:00   57:20
              6-MAY-88    Sir: Voltage 705    Sys: DB5US
  Sample 1 Injection 1    Group 4    Mass 423.7766
              1,2,3,4,6,7,9-HpCDD
 100,
  80-
  60-
  40
  20
                                      Norm:   282
                      1,2,3,4,6,7,8-HpCDD
     45:20   46:40   48:00   49:20   50:40   52:00   53:20   54:40   56:00   57:20
             6-MAY-88    Sir: Voltage 705     Sys: DB5US
  Sample 1 Injection  1    Group 4    Mass 441.7428
100
80
60
40'
20-
n.


' ' " I
•

^.I. . ...t. 	 ^. - 	 L - . -.-•-, 	 J
Norm: 1;
OCDF
z

I
I
V-L. 	 	 . ..
    45:20  46:40   48:00  49:20   50:40   52:00   53:20   54:40  56:00   57:20

             6-MAY-88    Sir: Voltage 705    Sys: DB5US
 Sample 1  Injection 1    Group 4    Mass 457.7377
100

 80

 60-

 40

 20
   45:20   46:40  48:00   49:20   50:40   52:00   53:20   54:40   56:00   57:20

         FIGURE 20  First and Last Eiuted Hepta- Dioxin and  Furan Isomers
                                                                                      15

-------
             3A   DB225 Column

                        21-APR-88    Sir: Voltage 705    Sys: DB225
             Sample 1 Injection 1    Group 1    Mass 305.8987
             Text: COLUMN PERFORMANCE
                                               2,3,7,8-TCDF      Norm:   3466
            100
             80
             60
             40
             20
                                   2,3,4,7-TCDF
y
I
                                                            1,2,3,9-TCDF
               16:10 16:20 16:30 16:40 16:50 17:00 17:10 17:20 17:30 17:40 17:50 18:00
                                                                                                       r
                                                                       t
                                                                       t
            3B   OB5 Column
            100
                                                                       I

                                                                       I
                                                                                                       I
                                                                                                       c
                                                                                                       c
                         22:30        24:00        25:30
                                           Time
                              27:00
  FIGURE 3  Valley between 2,3,7,8- Tetra Dioxin and Furan Isomers and Other Closely Eluted Isomers
16

-------
         both   the   primary   and  secondary  m/z's
         specified in Table 3, for each calibration
         standard, as follows:
             RR
                           (A
                              1
                             ,n
                                            l
                           (A,
         where,

         A   and A    are the areas  of  the primary
            and  secondary  m/z's  for  the unlabeled
            compound.

         A.   and A,   are the areas  of  the primary
          .
      'I
and  secondary m/z's
compound.
                                   fop  the  labeled
         C   is  the  concentration  of  the  labeled
            compound in the calibration standard.
         Cn is  the  concentration of  the unlabeled
            compound in the calibration standard.

7.5.3    To  calibrate  the  analytical  system  by
         isotope dilution,  inject a  1.0  uL aliquot
         of calibration standards  CS1 through  CSS
         (Section 6.13  and   Table  4)  using  the
         procedure in Section 13 and the  conditions
         in Table 2.   Compute the relative response
         (RR)  at each concentration.

7.5.4    Linearity -: If the  relative  response  for
         any compound  is   constant   (less  than  20
         percent coefficient  of  variation)  over  the
         5-point  calibration  range,  an  averaged
         relative response may  be   used  for  that
         compound; otherwise,  the complete  calibra-
         tion  curve for  that  compound shall  be used
         over  the 5-point calibration range.

  7.6    Calibration  by internal standard  --  The
         internal  standard  method  is applied   to
         determination   of  non-2,3,7,8-substituted
         compounds having no  labeled  analog  in this
         method,   and to  measurement  of   labeled
         compounds  for  intraLaboratory  statistics
         (Sections 8.4 and 14.5.4).

7.6.1    Response  factors  --  Calibration  requires
         the determination of  response  factors (RF)
         defined by the following equation:
        RF
Cis
                 
-------
          isotope     dilution)     and     multipoint
          calibration  curves.     Computations   of
          relative standard  deviation  (coefficient
          of   variation)   shall  be  used  to  test
          calibration  linearity.    Statistics   on
          initial   performance.  (Section   8.2)   and
          ongoing  performance  (Section 14.5)  shall
          be computed and maintained.

     8  '  QUALITY  ASSURANCE/QUALITY COMTROL
   8.1
 8.1.1
 8.1.2
 8.1.3
  8.1.4
Each  laboratory that uses  this method  is
required  to   operate  a  formal  quality
assurance  program  (Reference  16).    The
minimum   requirements  of   this  program
consist  of  an  initial  demonstration  of
laboratory capability,  analysis  of samples
spiked with  labeled compounds to evaluate
and document data quality, and analysis  of
standards and blanks  as tests of continued
performance.    Laboratory  performance  is
compared    to    established   performance
criteria  to determine  if the  results  of
analyses meet the performance characteris-
tics  of  the method.   If  the method is  to
be applied routinely  to samples  containing
high  solids  with  very   little moisture
(e.g., soils,  filter cake, compost) or  to
an  alternate   matrix,   the  high   solids
reference  matrix (Section 6.6.2) . or the
alternate  matrix (Section  6.6.4)  is  sub^
stituted  for   the   reagent  water   matrix
(Section 6.6.1)  in  all'performance tests.

The analyst shall  make an initial  demon-
stration   of   the   ability  to  generate
acceptable  accuracy   and precision  with
this  method.   This ability  is  established
as described in Section 8.2.

The  analyst is permitted  to  modify  this
method to  improve separations or lower the
costs  of measurements,  provided that all
performance  specifications are met.   Each
time  a modification is  made  to  the  method,
the analyst  is  required to repeat the pro-
cedures  in Sections  7.2  through  7.4 and
Section    8.2    to   demonstrate    method
performance.

Analyses  of blanks are required to  demon-
strate freedom from contamination (Section
3.2).    The procedures  and  criteria  for
analysis  of   a  blank  are  described  in
Section 8.5.

The   laboratory shall  spike  all  samples
with  labeled  compounds to monitor  method
performance.    This  test  is described  in
        Section 8.3.  When results of-these  spikes
        indicate  atypical  method  performance  for
        samples,  the  samples  are diluted to  bring
        method    performance   within    acceptable
        limits.    Procedures  for  dilutions,  are
        given-in  Section 16.4.

8.1.5   The  laboratory shall, on an ongoing  basis,
        demonstrate  through  calibration  verifica-
        tion and  the  analysis of the precision  and
        recovery  standard th.it the analytical sys-
        tem  is  in control.   These procedures  are
        described in  Sections 14.1 through 14.5.

8.1.6   The  laboratory shall maintain  recprds  to
        define  the quality  of data that  is  gener-
        ated.   Development, of accuracy  statements
        is described  in Section 8.4.       \

  8.2   Initial   precision   and   accuracy '.  —   To
        establish the ability to generate accept-
        able precision and  accuracy,,  the Analyst
        shall perform the following operations.

8.2.1   For  low solids  (aqueous  samples), extract,
        concentrate,   and   analyze  four  ,1-liter
        aliquots  of  reagent  water spiked with  the
        diluted   precision  and  recovery standard
        (PAR) (Sections 6.14  and 10.3.4)  according
        to  the  procedures in Sections  10  through
        13.   For an  alternate sample matrix, four
        aliquots  of  the alternate matrix  are used.
        All   sample   processing   steps, . including
        preparation    (Section    10),  '  extraction
        (Section   11),  and  cleanup  (Section  12)
        that are  to  be used  for  processing  samples
        shall be  included in  this test.

8.2.2   Using results of the  set  of four analyses,
        compute  the  average  concentration   (X)  of
        the  extracts  in ng/mL  and  the standard
        deviation  of  the   concentration .(s)   in
        ng/mL  for   each   compound,   by   isotope
        dilution   for  PCDDs  and  PCDFs   with  a
         labeled  analog, and  by   internal standard
        for   labeled  compounds.    Calculate  the
        recovery  of  the  labeled  compounds.

8.2.3    For  each  unlabeled  and   labeled  compound,
        compare  s  and  X  with  the  corresponding
         limits  for initial precision and accuracy
         in Table 7.  If s and X  for  all compounds
        meet  the   acceptance   criteria,   system
         performance is acceptable, and analysis of
         blanks    and  samples  may  begin.      If,
         however,   any  individual   s   exceeds  the
         precision limit or any  individual  X falls
         outside  the  range   for  accuracy,   system
18
                                                                                                                                 c
                                                                                                                                 I

-------
        performance   is   unacceptable   for  that
        compound.  Correct  the  problem and; repeat
        the test (Section 8.2).  The concentration
        limits  in  Table  7 for  labeled compounds
        are  based  on  the  requirement that   the
        recovery  of  each  labeled compound be  in
        the range of 25-150%.

  3.3   The laboratory shall, spike all  samples-and
        QC  aliquots   with . the  diluted   labeled
        compound  spik:ng  solution  (Sections  6.10
        and  10.3.2)  to  assess  method  performance
        on the  sample matrix.
8.3.1
8.3.2
8.3.3
   8.4
 8.4.1
 8.4.2
Analyze  each  sample   according  to   the
procedures in Sections 10 through 13.

Compute  the  percent  recovery  (R)  of  the
labeled  compounds  in the labeled compound
spiking  standard  and the cleanup standard
using    the    internal   standard    method
(Section 7.6).

The recovery of each  labeled compound must
be within 25-150%.   If  the  recovery  of any
compound falls outside of  these  limits,
method   performance   is unacceptable  for
that compound  in  that  sample.   To  overcome
such   difficulties,   water  samples   are
diluted   and   smaller   amounts   of   soils,
sludges, sediments and other matrices are
reanalyzed per Section 17.

Method  accuracy  for  samples  shall  be
 assessed and  records shall  be maintained.

After  the analysis of  five samples  of a
 given  matrix  type  (water,  soil,   sludge,
 pulp,  etc)  for which  the  labeled compound
 spiking  standards   pass    the  tests  in
 Section 8.3,  compute  the  average   percent
 recovery (R)  and the standard  deviation of
 the percent recovery  (SR)  for  the  labeled
 compounds  only.     Express  the  accuracy
 assessment as a  percent recovery interval
 from R
                   2SR  to  R
                     2SR  for each matrix.
For example," if R  = 90K and  SR  = 10% for
five  analyses   of  pulp,   the  accuracy
interval is expressed as 70-110%.

Update  the accuracy  assessment  for  each
compound in each matrix on a  regular basis
(e.g.,   after   each  5-10   new  accuracy
measurements).
    8.5    Blanks  --  Reference  matrix  blanks  are
          analyzed   to   demonstrate   freedom  from
          contamination (Section 3.2).
8.5.1   Extract  and  concentrate a  1-liter  reagent
        water  blank   (Section  6.6.1),  high  solids
        reference  matrix  blank  (Section  6.6.2),
        paper  matrix  blank  (Section  6.6.3)   or
        alternate  reference matrix  blank  (Section
        6.6.4)   with  each  sample  set  (samples
        started  through the extraction  process  on
        the  same 12-hour shift, to a maximum of  20
        samples).    Analyze the blank  itnnediately
        after   analysis   of   the   precision  and
        recovery  standard   (Section  14.5)   to
        demonstrate  freedom from contamination.

8.5.2    If any of the  PCDOs or PCDFs  (Table 1)  or
        any potentially  interfering  compound  is
         found  in blank at greater than  the minimum
         level   (Table   2),  assuming   a  response
         factor of 1  relative to the   C12-1,2,3,4-
         TCDD internal  standard for  compounds  not
         listed in Table 1,  analysis of samples is
         halted until  the  source  of  contamination
         is  eliminated  and   a   blank   shows   no
         evidence  of contamination  at  this  level.
         NOTE:     All   samples  associated  with  a
         contaminated  method   blank   must  be   re-
         extracted   and   reanalyzed   before   the
         results  may  be  reported  for   regulatory
         compliance  purposes.

   8.6   The   specifications   contained  in  this
         method can  be  met  i.f  the  apparatus  used is
         calibrated  properly and  then  maintained in
         a "calibrated  state.    The standards used
         for  calibration  (Section 7),  calibration
         verification   (Section   14.3),  and   for
         initial  (Section 8.2) and ongoing (Section
         14.5)  precision  and  recovery should  be
         identical,   so   that  the   most   precise
         results  will be obtained.  A GCHS  instru-
         ment  will  provide  the  most   reproducible
         results if  dedicated to the  settings  and
         conditions   required   for  the  analyses  of
         PCODs  and PCDFs by this method.
                                         \
    8.7   Depending  on  specific  program ^-require-
         ments,  field  replicates may be collected
          to determine the precision of  the sampling
          technique,   and   spiked   samples  may   be
          required to determine the accuracy of  the
          analysis when  the  internal standard method
          is used.
                                                               9    SAMPLE   COLLECTION,
                                                                    HANDLING
                                                                                           PRESERVATION,   AND
                                                     9.1   Collect  samples in amber  glass  containers
                                                           following  conventional  sampling  practices
                                                           (Reference  17).    Aqueous  samples  which
                                                           flow freely are collected in refrigerated
                                                                                                            19

-------
           bottles  using  automatic  sampling  equip-
           ment.  Solid samples are collected as grab
           samples using wide mouth jars.

    9.2    Maintain  samples  at 0-4  °C  in the  dark
           from   the    time   of   collection   until
           extraction.     If  residual  chlorine  is
   ~*       present  in   aqueous  samples,  add  80  mg
           sodium  thiosulfate  per liter  of  water.
           EPA Methods  330.4  and 330.5  may be u >ed to
           measure residual chlorine (Reference 18).

    9.3    Perform sample, analysis within 40  days of
           extraction.

     10    SAMPLE PREPARATION

           The  sample  preparation  process  involves
           modifying the physical   form of  the sample
           so  that   the  PCDOs  and   PCOFs   can  be
           extracted  efficiently.     In general,  the
           samples must  be  in a  liquid  form or in the
           form of finely divided solids  in order for
           efficient extraction  to  take place.  Table
           8   lists   the   phase(s)   and   quantity
           extracted  for   various   sample matrices.
           Samples  containing   a   solid   phase   and
           samples  containing  particle sizes  larger
           than  1  im   require  preparation  prior' to
           extraction.      Because   PCODs/PCOFs   are
           strongly associated  with particulates,  the
           preparation  of  aqueous  samples is  depen-
           dent on  the  solids content  of  the  sample.
           Aqueous  samples  containing  less than  one
           percent   solids   are   extracted   in  a
           separator/   funnel.     A  smaller   sample
           aliquot   is   used   for   aqueous   samples
           containing  one  percent  solids  or more.
           For samples  expected or known to  contain
           high levels of the PCOOs and/or PCDFs,  the
           smallest sample size  representative of  the
           entire  sample  should  be  used,  and  the
           sample  extract   should  be  diluted,   if
       '   necessary, per Section 16.4.

   10.1   Determine percent solids

 10.1.1   Weigh 5-10 g of  sample  (to  three  signifi-
           cant figures).into a tared  beaker.  N01£:
           This aliquot  is  used only for  determining
           the solids content of the sample,  not  i'or
           analysis of PCODs/PCDFs.

 10.1.2   Dry overnight (12  hours  minimum) at 110 ±5
           •C, and cool  in a dessicator.
10.1.3   Calculate percent solids as follows:
                                         x 100
         % solids =
         weight of sample after drying
         weight of sample before drying

  10.2   Determine particle size

10.2.1   Spread   the  dried  sample  from  Section
         10.1.2  on  a  piece  of  filter paper  or
         aluminum foil  in a fume hood or glove box.

10.2.2   Estimate the  size  of the particles in  the
         sample.     If  the   size  of  the  largest
         particles   is  greater   than  1  mm,   the
         particle size  must be reduced  to  1  mm or
         less prior  to  extraction.         :

  10.3   Preparation' of aqueous  samples containing
         one   percent   solids   or   less  '••    The
         extraction  procedure for  aqueous • samples
         containing  less  than  or  equal  to   one
         percent  solids   involves  filtering   the
         sample,  extracting  the  particulate  phase
         and the filtrate separately, and combining
         the  extracts   for  analysis.    The  aqueous
         portion  is  extracted   by  shaking  with
         methylene chloride in a separatory funnel.
         The  particulate   material   is  extracted
         using the SOS  procedure.          ;

10.3.1   Mark the  original  level of  the sample on
        - the  sample, bottle  for  reference.    Weigh
         the sample  in  the  bottle  on
-------
10.3.5
        standard  (Section  6.14)  to  2.0  mL  with
        acetone.    Spike  1.0  mL of  the  diluted
        precision  and  recovery standard  into  .the
        remaining   reagent' water aliquot.    This
        aliquot  will  serve  as  the  PAR  (Section
        14.5).'     '  '

        Assemble  a Buchner  funnel  on  top of  a
        clean  1  L  filtration  flask.    Apply  a
        vacuum to  the  flask,  and  pour  the entire
        contents  of  the  sample bottle  through  a
        glass fiber filter  (Section 5.5.4) in the
         Buchner   funnel,   swirling   the  sample
         remaining  in  the bottle  to  suspend  any
         particulates.

         Rinse the  sample bottle twice with  5 mL  of
         reagent  water  to transfer  any  remaining
         particulates onto the  filter.

         Rinse  the  any  particulates off  the  sides
         of  the Buchner  funnel with  small  quanti-
         ties of reagent water.

         Weigh  the  empty sample  bottle on a  top-
          loading  balance  to ±1  g.    Determine the
         weight  of  the  sample by difference.   Oo
          riot discard the bottle at this point.

 10.3.9    Extract  the filtrates using  the procedures
          in Section 11.1.1.

10.3.10    Extract  the particulates using the proce-
          dures in Section 11.1.2.

   10.4    Preparation of samples containing greater
          than one percent solids

 10.4.1    Weigh a well-mixed aliquot  of each sample
          (of  the   same  matrix  type)  sufficient  to
          provide 10 g  of dry  solids (based on  the
          solids  determination  in  10.1.3)  into a
          clean beaker  or glass jar.

 10.4.2   Spike   1.0  mL  of   the  diluted  labeled
          compound  spiking solution (Section 10.3.2)
           into  the  sample aliquot(s).
10.3.6
 10.3.7
 10.3.8
  10.4.3
  10.4.4
                                      i
           For  each  sample  or  sample  set  (to  a
           maximum  of  20  samples) ' to  be  extracted
           during the  same 12-hour  shift,  weigh two
           10 g aliquots of the appropriate reference
           matrix (Section 6.6) into clean beakers or
           glass jars.

           Spike  1.0  mL   of  the  diluted   labeled
           compound   spiking   solution   into   one
           reference  matrix  aliquot.    This  aliquot
           will serve  as  the blank.  Spike 1.0 mL-of
           the   diluted    precision   and    recovery
         standard   (Section   10.3.4)   into   the
         remaining reference  matrix  aliquot.   This
         aliquot  will  serve  as  the  PAR  (Section
         14.5).

10.4.5   Stir   or   tumble   and   equilibrate  the
         aliquots  for  1-2  hours.

10.4.6   Extract  the aliquots using  the procedures
         in  Section  11.

  10.5   Multiphase  samples

10.5.1   Pressure filter the  sample,  blank,  and PAR
         aliquots through Whatman GF/0 glass  fiber,
         filter paper.    If  necessary, centrifuge
         these aliquots  for  30 minutes at  greater
          than 5000 rpm prior  to filtration.

10.5.2   Discard  any aqueous  phase  (if  present).
          Remove any non-aqueous liquid (if present)
          and  reserve  for recombination  with  the
          extract  of   the   solid   phase   (Section
          11.1.2.5').   Prepare the filter  papers of
          the  sample  and  QC  aliquots  for, particle
          size   reduction  and   blending   (Section
          10.6).

   10.6   Sample grinding, homogenization, or  blend-
          ing  .. samples  with particle sizes greater
          than   1   mm  (as   determined  by  Section
          10.2.2)  are subjected  to   grinding,  homo-
          genization, " or  blending.    The  method of
          reducing particle   size to  less  than .1 mm
           is matrix  dependent.   In general, .hard
          particles  can be reduced by grinding  with
           a  mortar and pestle.   Softer particles  can
          be reduced by grinding in  a Wiley mill  or
           meat .grinder,   by  homogenization,  or  by
           blending.

  10.6.1    Each  size   reducing  preparation  procedure
           on each matrix shall be verified by run-
           ning  the  tests in  Section  8.2  before  the
           procedure is employed routinely.
                                              V
  10.6.2   The  grinding,  homogenization, or  blending
           procedures  shall be carried out  in a glove
           box or  fume hood to prevent particles from
           contaminating the  work environment.

  10.6.3   Grinding -- Tissue samples, certain papers
           and  pulps,  slurries,  and  amorphous solids
           can  be ground  in  a  Wiley mill  or heavy
           duty meat  grinder.  In some cases,  reduc-
           ing   the   temperature  of   the  sample  to
           freezing  or to dry ice or liquid nitrogen
           temperatures   can   aid  in  the  grinding
           process.    Grind the sample aliquots  from
           Section  10.4.5  or  10.5.2  in  a  clean
                                                                                                             21

-------
          grinder.  Do  not  allow the sample tempera-
          ture to exceed 50 "C.   Grind the blank and
          reference  matrix  aliquots  using a  clean
          grinder.

10.6.4    Homogeniza'tion or  blending  --  Particles
=.         that  are  not  ground   effectively,   or
          particles  greater than 1  mm in size after
          grinding,  can  ften be reduced in size by
         •high  speed  himogenization  or  blending.
          Homogenize  and/or blend the sample, blank,
          and  PAR   aliqMots  from  Section  10.4.5,
          10.5.2, or  10.£.3.

10.6.5    Extract  the aliquots  using  the procedures
          in Section  11.

     11    EXTRACTION  AND CONCENTRATION

   11.1 .  Extraction  of filtrates  --   extract  the
        " -aqueous  samples,  blanks,  and PAR aliquots
          according  to  the  following procedures.

11.1.1    Pour the filtered aqueous sample from the
          filtration flask  into a  2-L separatory
          funnel.   Rinse, the flask twice  with 5 mL
          of  reagent water and add these  rinses to
          the separatory funnel.   Add  60  mL methy-
          lene  chloride  to   the   sample  bottle
          (Section   10.3.8),    seal,and   shake  60
          seconds to rinse the inner surface.

 11.1.2   Transfer  the  solvent  to  the  separatory
          funnel and extract the sample by  shaking
          the  funnel  for  2  minutes  with periodic
          venting.    Allow  the  organic   layer  to
          separate  from  the   water   phase  for   a
          minimum of  10 minutes.   If the emulsion
          interface between layers  is more than one-
          third  the volume of  the  solvent  layer,
          employ mechanical techniques  to complete
          the phase  separation  (e.g., a glass  stir-
          ring  rod).    Drain  the methylene  chloride
          extract into  a solvent-rinsed glass funnel
          approximately one-half  full ' of   clean
          sodium sulfate.   Set  up  the  glass  funnel
          so that  it   will  drain  directly  into  a
          solvent-rinsed  StlO-mL  K-D   concentrator
          fitted  with   a  10  mL  concentrator  tube.
          NOTE:    Experience  with  aqueous  samples
          high  in dissolved organic materials (e.g.,
          paper mill effluents)  has shown that acid-
          ification    of   the    sample   prior   to
          extraction  may  reduce  the  formation  of
          emulsions.   Paper industry  methods suggest
          that  the  addition  of up   to 400  mL  of
          ethanol to a  1 L effluent  sample may also
          reduce   emulsion   formation.     However,
          studies by the Agency to  date suggest that
          the effect may be a  result  of the dilution"
11.1.4
         of the  sample, and  that  the  addition of
         reagent water may serve  the same function.
         Mechanical techniques  may still be neces-
         sary to complete the phase separation.  If
         either  of these  techniques  is utilized,
         the  laboratory must  perform  the startup
         tests described  in Section  8.2 using  the
         same techniques.

11.1.3   Extract  the  water  sample two  more  times
         using  60 mL  of fresh  methylene  chloride
         each time.  Drain each extract  through  the
         funnel  containing  the sodium sulfate  into
         the  <-D  concentrator.    After  the  third
         extraction,  rinse  the  separatory funnel
         with  at  least  20  mL of  fresh methylene
         chloride, and drain this rinse  through  the
         sodium  eulfate   into   the   concentrator.
         Repeat  this rinse at  least twice.

         The  extract   of   the   filtrate   must  be
         concentrated  before  it   is  combined  with
         the   extract   of   the  particulates   for
         further cleanup.    Add  one  or two  clean
         boiling chips  to the  receiver and  attach  a
         three-ball  macro  Snyder column.    Prerwet
         the column by  adding  approximately 1  mL of
         hexane  through the  top,.    Place  the  K-D
         apparatus in  a hot water bath  so  that  the
         entire  lower  rounded surface of  the  flask
         is bathed with steam.

11.1.5   Adjust   the   vertical'  position  of   the
         apparatus and  the  water  temperature as,
         required to  complete  th«  concentration in
         15-20  .minutes.    At   the proper  rate of
         distillation,  the  balls  of the|column will
         actively chatter but  the chambers will  not
         flood.

11.1.6   When  the  liquid  has  reached ;an  apparent
         volume of 1 mL,  remove  the  K-D  apparatus
         from  the bath and  allow  the  solvent to
         drain   and cool  for  at  least  10  minutes.
         Remove  the  Snyder  column  and rinse  the
         flask  and its  lower joint into the concen-
          trator tube  with  1-2  mL  of hexane.  A 5 mL
         syringe is recommended for this operation.

11.1.7   The concentrated extracts of the filtrate
          and the  particulates  are  combined  using
          the procedures in Section 11.2.13.

   11.2    Soxhlet/Dean-Stark extraction lof  solids --
          Extract  the  solid samples,  particulates,
          blanks, and PAR aliquots using the follow-
          ing procedure.

 11.2.1    Charge a clean extraction thimble with 5.0
          g of 100/200 mesh silic
-------
           and  100  g of quartz sand  (Section 6.3.2).
           NOTE:    Do not  disturb  the  silica  layer
           throughout the extraction  process.

 11.2.2    Place  the  thimble  in  a   clean  extractor.
           Place'  30-40 ml  of toluene in  the receiver
           and  200-250 mL of toluene  in the  flask.

 11.2.3    Pre-extract the glassware by  heating  the
           flask  until the toluene is boiling.   When
           properly adjusted,   1-2 drops  of  toluene
           per  second will  fall   from the  condenser
           tip  into  the   receiver.    Extract   the
           apparatus for three hours  minimum.

 11.2.4    After  pre-extraction,  cool and  disassemble
           the  apparatus.    Rinse the  thimble  with
           toluene  and allow to air dry.

 11.2.5    Load the wet  sample from  Sections  10.4.6,
           10.5.2,  10.6.3,  or  10.6.4, and  any  non-
           aqueous   liquid  from   Section  10.5.2  into
           the  thimble and manually mix into the  sand
           layer  with  a clean metal  spatula  carefully
           breaking  up any  large  lumps of  sample.   If
           the  material   to   be   extracted   is   the
           particulate matter  from the filtration  of
           an aqueous  sample, add the filter paper  to
        •   the  thimble also.

 11.2.6    Reassemble  the pre-extracted SOS  apparatus
           and  add  a  fresh charge of toluene to  the
           receiver  and reflux flask.

 11.2.7    Apply  power to the heating mantle to begin
           ref lu'xing.    Adjust  the   reflux   rate  to
           match  the rate  of  percolation through  the
           sand and silica beds  until  water  removal
           lessens  the restriction  to  toluene flow.
           Check  the apparatus for foaming frequently
           during  the first 2 hours of  extraction.
           If foaming  occurs,  reduce the reflux  rate
           until  foaming subsides.

 11.2.8    Drain  the water from  the  receiver at 1-2
           hours  and  8-9  hours,  or  sooner  if  the
          'receiver  fills  with   water.    Reflux  the
           sample for a  total  of  16-«24 hours.  Cool
           and disassemble the apparatus.   Record the
           total volume of water collected.

 11.2.9    Remove  the distilling  flask.   Drain  the
           water  from  the Dean Stark  receiver  and add
          'any toluene in the receiver to the  extract
           in the flask.

11.2.10    For  solid  samples,  the  extract   must  be
           concentrated to  approximately  10  mL prior
           to back  extraction. -  For   the particulates
           filtered   from  an  aqueous  sample,   the
           extract   must  be  concentrated  prior  to
           combining  with    the   extract   of   the
           filtrate.   Therefore,  add one or two clean
           boiling  chips  to  the  round bottom flask
           and   attach  a   three-ball   macro  Snyder
           column.    Pre-wet  the  column  by  adding
           approximately 1 mL of  toluene  through the
           top.   Place  the round  bottom  flask  in  a
           heating  mantle and apply heat  as  required
           to  complete  the  concentration  in  15-20
           minutes.   At  the proper  rate of  distilla-
           tion,   the  balls   of   the  column  will
           actively chatter  but  the chambers  will not
           flood.                 ''

11.2.11    When  the  liquid  has  reached an  apparent
           volume  of 10 mL,  remove the round  bottom
           flask  from the heating  mantle and  allow
           the solvent to  drain  and cool for  at least
           10 minutes.   Remove the  Snyder  column.

11.2.12    If the ex'tract  is  from  a solid  sample,  not
           the  particulates from  an aqueous  sample,
           transfer  the  concentrated extract  to a 250
           mL  separatory  funnel.    Rinse  the  flask
           with  toluene  and  add   the   rinse  to  the
           separatory  funnel.    Proceed  with  back
           extraction per  Section 11.3.

11.2.13    If  the  extract is  from the particulates
           from  an aqueous  sample,  it  must  be  com-
           bined with the  concentrated  extract  of  the
           filtrate  (Section  11.1.7)  prior  to  back
           extraction.    Assemble  the  glass  funnel
           filled  approximately  one-half   full  with
           sodium  sulfate  from Section  11.1.2  such
           that  the funnel  will  .drain  into  the  K-0
           concentrator  from Section 11.1.7  contain-
           ing  the  concentrated  methylene   chloride
           extract of the  filtrate.  Pour  the concen-
           trated toluene  extract of the particulates
           through  the sodium  sulfate  into  the  K-D
          concentrator.      Rinse   the  round-bottom
           flask  with  three  15-20  mL  volumes  of
          hexane,   and  pour  each  rinse  through  the
          sodium sulfate  into the <-D  concentrator.
          Add one or  two  fresh  boiling chips  to  the
           receiver  and  attach  the  three-ball  macro
          Snyder  column  to  the   K-D   concentrator.
          Pre-wet  the column by adding  approximately
          1 mL of  hexane to  the  top  of the column.
          Concentrate   the   cjpmbined  extract   to
          approximately  10  ml  
-------
         and lower joint with  three 5 ml  volumes  of
         hexane, and  add  each rinse to the  separa-
         tory funnel.  Proceed with back  extraction
         pep Section  11.3.

  11.3   Back extraction with  base and acid
14.3.1
11.3.2
         Spike  1.0  mL  of  the  cleanup  standard
         (Section 6.11) into the separatory funnels
         containing  the  sample  and  QC  extracts
         (Section 11.2.12 or 11.2.13).

         Partition  the extract  against  50  mL  of
         potassium   hydroxide   solution  (Section
         6.1.1).  Shake for 2 minutes with periodic
         venting into  a hood.   Remove  and discard
         the aqueous layer.   Repeat the base wash-
         ing  until  no  color  is   visible  in   the
         aqueous layer, to  a  maximum of four wash-
       ,  ings.   Minimize contact  time  between  the
        ^extract and  the base  to  prevent degrada-
         tion  of  the  PCOOs  and PCDFs.   Stronger
         potassium   hydroxide   solutions   may  be
         employed  for  back  extraction,  provided
         that  the  laboratory meets the specifica-
         tions  for  labeled  compound  recovery  and
         demonstrates  acceptable performance using
         the procedures in Section 8.2.

11.3.3   Partition  the extract against  50  mL of
         sodi.um  chloride  solution  (Section  6.1.3)
         in the  same way as with base.   Discard  the
         aqueous layer.

11.3.4   Partition  the  extract against  50  mL of
         sulfuric  acid (Section 6.1.2)  in the  same
         way as  with base.  Repeat  the  acid  washing
         until  no  color is visible in  the  aqueous
         layer,  to a maximum of  four washings.

11.3.5   Repeat  the   partitioning   against   sodium
         chloride  solution  and discard  the  aqueous
         layer.

11.3.6   Pour  each extract  through a drying  column
         containing  7  to 10 cm  of  anhydrous  sodium
         sulfate.  Rinse the  separatory funnel  with
         30-50 mL  of  toluene  and pour  through  the
         drying  column.  Collect each  extract  in  a
         500  mL round bottom  flask.    Concentrate
         and  clean up  the  samples and  QC aliquots
         per  Sections  11.4  and  12.

  11.4   Macro-concentration   --   Concentrate   the
         extracts  in  separate  100-mL  round  bottom
         flasks  on a rotary evaporator.

11.4.1   Assemole  the  rotary  evaporator  according
         to manufacturer's  instructions,  and  warm
         the  water  bath  to  45  °C.    On a  daily
         basis,  preclean  the  rotary  evaporator by
         concentrating  100  mL  of  clean extraction
         solvent through  the  system.   Archive both
         the  concentrated solvent and  the solvent
         in the catch flask for contamination check
         if necessary.   Between samples,  three 2-3
         mL  aliquots of  toluene should  be  rinsed
         down the  feed tube into a waste beaker.

11.4.2   Attach  the round  bottom flask containing
         the  sample extract to the rotary evapora-
         tor.   Slowly apply  vacuum  to  the system,
         and begin rotating the sample flask.

11.4.3   Lower  the  flask  into the water  bath and
         adjust  the speed   of   rotation  and  the
         temperature as  required to complete the
         concentration  in  15-20  minutes.    At the
         proper  rate of concentration,  the flow of
        • solvent  into the  receiving  flask will be
         steady, but no bumping or visible boiling
         of  the  extract will  occur.   NdTE:  If the
         rate of concentration is>  too fast, analyte
         loss may  r-ccur.

11.4.4   When the  liquid  in the concentration  flask
         has  reached an  apparent volume  of  2 mL,
         remove  the flask  from the  water bath and
         stop the  rotation.   Slowly and  carefully,
         admit air into the system.  Be sure not  to
         open the  valve so quickly that  the sample
         is  blown  out of  the  flask.  Rinse the feed
         ' tube with approximately  2 mL of hexane.

11.4.5   Transfer  the extract to  a vial using  three
         2-3 mL  rinses  of  hexane.    Proceed  with
         micro-concentration  and  solvent  exchange.

   11.5   Micro-conc°ntration  and  solvent  exchange

11.5.1   Toluene extracts to be subjected to  GPC  or
         HPLC cleanup are  exchanged  into methylene
         chloride.  Extracts  that are  to  be  cleaned
         up using  silica gel, alumina!,  and/or  AX-
         21/Celite are  exchanged into hexane.

11.5.2   Transfer   the  vial  containing the  sample
         extract  to a  nitrogen evaporation  device.
         Adjust  the flow of  nitrogen  so that  the
          surface  of the  solvent is   just  visibly
         disturbed.  MOTE:   A large vortex  in the
          solvent may cause analyte loss.

11.5.3    Lower the vial into a 45 °C water bath and
          continue  concentrating.

11.5.4    When the  volume of  the liquid is approxi-
          mately 100  uL,  add Z-3 mL of the  desired
          solvent  (methylene chloride or hexane) and
                                                                                                                            r
                                                                                                                             c
                                                                                                                             r
                                                                                                                             r
24

-------
           continue  concentration  to   approximately
           100  uL.   Repeat  the  addition  of  solvent
           and concentrate once more.

  11.5.5   If the extract  is  to be cleaned up by GPC
           or HPLC, adjust  the  volume of the  extract
           to   5.0   ml   with   methylene   chloride.
           Proceed with GPC cleanup (Section 12.2).

  11.5.6   If  the  extract  is  to be  cleaned up  by
           column  chromatography   (alumina,   silica
           gel,  AX-21/Celite), bring the final volume
           to  1.0  mL  with   hexane.    Proceed  with
           column cleanups (Sections 12.3-12.5).

  11.5.7   For  extracts   to   be   concentrated   for
           injection  into  the GCHS --  add 10 uL  of
           nonane to  the  vial.   Evaporate the solvent
           to the level of the nonane.   Evaporate the
           hexane in the vial  to the  level   of  the
           nonane.

  11.5.8:   Seal  the vial  and  label with the  sample
           number.  Store in the dark  at  room temper-
           ature  until ready for  GCHS analysis.

     12    EXTRACT CLEANUP

   12.1    Cleanup   may   not    be   necessary  for
           relatively  clean samples  (e.g.,   treated
           effluents,   groundwater,  drinking   water).
           If  particular   circumstances   require  the
           use of a cleanup  procedure,  the   analyst
          may use any or all of  the procedures below.
        I  or   any   other  appropriate   procedure.
          Before  using  a  cleanup  procedure,  the
          analyst must  demonstrate that the require-
          ments of  Section 8.2  can  be met  using  the
          cleanup procedure.
 12.1.1
          Gel   permeation  chromatography   (Section
          12.2)  removes  many  high molecular  weight
          interferences    that    cause   GC   column
          performance  to  degrade.    It  may be  used
          for  all  soil and sediment extracts  and may
          be   used  for   water  extracts   that   are
          expected to  contain high molecular  weight
          organic    compounds    (e.g.,     polymeric
          materials, humic  acids).
12.1.2  . Acid,  neutral,  and basic  silica gel, and
         alumina  (Sections  12.3  and 12.4) are used
         to     remove    nonpolar     and    polar
         interferences.

12.1.3   AX-21/Celite  (Section  12.5)   is used  to
         remove nonpolar interferences.
   12.1.4    HPLC  (Section  12.6)  is  used  to  provide
             specificity  for  the  2,3,7,8-substituted
             and other  PCDO  and PCDF  isomers.

     12.2    Gel permeation  chromatography (GPC)

   12.2.1    Column packing

 12.2.1.1
            Place  70-75  g of SX-3 Bio-beads  in  a 400-
            500 mL beaker.

            Cover  the beads with  methylene  chloride
            and  allow  to swell  overnight  (12  hours
            minimum).

            Transfer  the swelled beads  to the  column
            and pump  solvent through the column,  from
            bottom to  top,  at 4.5-5.5 mL/min prior  to
            connecting the column to the detector.
                          •
            After purging  the column with solvent for
            1-2 hours, adjust the column head pressure
            to 7-10  psig and purge  for  4-5  hours  to
            remove air.   Maintain a  head pressure  of
            7-10 psig.    Connect  the  column to the
            detector.
  12.2.2    Column calibration
12.2.2.1
 12.2.1.2
 12.2.1.3
 12.2.1.4
12.2.2.2
12.2.2.3
            Load  5  mL  of   the  calibration  solution
            (Section 6.4)  into the sample loop.

            Inject the calibration solution and record
            the signal  from the detector.  The elution
            pattern   will   be  corn  oil,  tjis(2-ethyl
            hexyl)    phthalate,     pentachlorophenol,
            perylene,  and  sulfur.

            Set  the  "dump  time"  to allow  >85  percent
            removal  of  the  corn oil  and  >85  percent
            collection of  the phthalate.

12.2.2.4 '   Set  the  "collect  time"  to  the peak minimum
           between perylene  and sulfur.

12.2.2.5   Verify the 'calibration  with  thevcalibra-
           tion  solution  after  every  20 extracts.
           Calibration is verified  if  the  recovery of
           the  pentachlorophenol  is  greater  than 85
           percent.    If calibration is not verified,
           the system shall be recalibrated using the
           calibration solution, and  the previous 20
           samples shall be  re-extracted and cleaned
           up using  the calibrated GPC system.

 12.2.3    Extract cleanup --  GPC  requires  that  the
           column not  be  overloaded.    The  column
           specified in  this method  is  designed  to
           handle a  maximum  of   0.5  g  of   high
           molecular   weight   material   in  a   5   mL •
                                                                                                          25

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           extract.    If  the  extract   is  known  or
           expected to  contain more than 0.5  g, the
           extract is split into aliquots for GPC and
           the  aliquots are  combined  after  elution
           from the  column.    The  residue content of
           the     extract     may    be    obtained
           gravirnetrically by  evaporating the solvent
           from a 50 uL aliquot:
12.2.3.1   Filter  the  extract
           filter  holder   to
           Load the 5.0 mL extract onto the column.
                      or  load  through  the
                      remove   particulates.
12.2.3.2   Elute  the  extract  using  the  calibration
           data   determined    in    Section   12.2.2.
           Collect  the eluate  in  a clean  400-500 mL
           beaker.
12.2.3.3
12.2.3.4
12.2.3.5
 Rinse the  sample loading  tube thoroughly
 with methylene  chloride between  extracts
%to prepare for the next sample.

 If   a   particularly   dirty   extract   is
 encountered, a  5.0 mL  methylene  chloride
 blank shall be  run through  the  system to
 check for carry-over.

 Concentrate the eluate per Section 11.2.1,
 11.2.2, and 11.3.1  or  11.3.2  for further
 cleanup or for injection into the GCHS.
  12.3   Silica gel cleanup

12.3.1   Place a  glass wool  plug in a  15  mm i.d.
         ehromatography column.   Pack the column in
         the following order  (bottom to top):  1 g
         silica gel (Section 6.5.1.1), four g basic
         silica gel  (Section 6.5.1.3),  1  g  silica
         gel,  8   g   acid  silica   gel  (Section
         6.5.1.2), 2 g  silica gel.  Tap the  column
         to settle the adsorbents.

12.3.2   Pre-rinse  the  column with  50-100  mL  of
         hexane.    Close   the  stopcock  when   the
         hexane   is  within  1  mm  of  'the   sodium
         sulfate.   Discard the eluate.   Check  the
         column for channeling.   If channeling  is
         present,  discard  the column  and prepare
         another.

12.3.3   Apply  the concentrated  extract   to   the
         column.    Open  the  stopcock  until   the
         extract  is  within  1  mm  of  the  sodium
         sulfate.

12.3.4   Rinse   the  receiver   twice  with  1   mL
         portions of hexane and apply separately to
         the column.   Elute  the PCDDs/PCDFs  with
         100 mL hexane and collect the eluate.
12.3.5   Concentrate the eluate per Section 11.4 or
         11.5  for  further  cleanup or for  injection
         into  the  HPLC or GCHS.         i

12.3.6   For  extracts  of samples  known to contain
         large  quantities   of   other   organic
         compounds (such as  paper mill effluents)
         it   may  be  advisable  to  increase  the
         capacity  of the silica  gel  column.   This
         may   be  accomplished  by  increasing  the
         strengths of  the  acid and  basic  silica
         gels.    The  acid   silica  gel   (Section
         6.5.1.2)  may be  increased  in .strength  to
         as  much  as  44% w/w  (7.9 g sulfuric acid
         added  to 10 g  silica  gel).    The basic
         silica   gel  (Section   6.5.1.3)   may   be
          increased in strength  to as  much  as 33%
         w/w  (50.mL 1H  HaOH added to  100  g  silica
         gel).  NOTE:    The  use  of stronger acid
         silica gel (44% w/w) may lead to charring
         of   organic compounds  in  some  extracts.
         The   charred  material may  retain some  of
          the  analytes and  lead  to lower recoveries
          of  PCDDs/PCOFs.   Increasing  the  strengths
          of  the acid and basic  silica  gel  may also
          require  different  volumes  of  hexane than
          those  specified   above,  to  elute   the
          analytes off the  column.   Therefore,  the
          performance  of   the  method   after  such
          modifications  must  be  verified  by  the
          procedures in Section' 8.2.

   12.4   Alumina  cleanup

 12.4.1   Place  a  glass  wool  plug in a 15 mm  i.d.
          ehromatography column.

 12.4.2   If using acid  alumina,  pack the column by
          adding 6 g acid alumina  (Section 6.5.2.1).
          If  using  basic  alumina,  substitute  6  g
          basic  alumina  (Section 6.5.2.2).   Tap the
          column to settle the adsorbents.

 12.4.3   Pre-rinse  the column  with  50-100  mL  of
          hexane.    Close   the  stopcock  when  the
          hexane is within 1 mm of the alumina.

 12.4.4   Discard  the  eluate.   Check the column for
          channeling.    If  channeling  is present,
          discard  the column and prepare another.

 12.4.5   Apply the concentrated extract-  to  the
          column.    Open   the  stopcock  until  the
          extract  is within  1  mm of the ^alumina.
                                                                                                                    r
                                                             12.4.6
                                                           Rinse  the  receiver   twice  .with  1   mL
                                                           portions  of  hexane and apply  separately  to
                                                           the  column.      Elute  the   interfering
                                                           compounds with  100  mL hexane and  discard
                                                           the eluate.
                                                                                                                    i
t
i
r
                                                                                                                               E
                                                                   C
                                                                   r
  26

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  12.4.7    The choice of eluting solvents will depend
         ',   on  the choice  of  alumina (acid  or basic)
            made  in Section 12.4.2.

12.4.7.1    If using acid alumina, elute the PCODs and
            PCOFs  from the column with 20 mL methylene
            chlorideihexane (20:80 v/v).   Collect the
            eluate.

12.4.7.2    If  using   basic alumina, elute the  PCOOs
            and  PCOFs  from  the  column  uith  20  mL
           methylene   chloride:hexane   (50:50   v/v).
           Collect the eluate.

  12.4.8   Concentrate the eluate per Section 11.4  or
           11.5 for further  cleanup or for  injection
           into the HPLC or GCMS.

    12.5   AX-21/Celite

  12.5.1    Cut both   ends from  a  10   ml  disposable
           serological  pi pet  to  produce  a  10   cm
           column.  Fire polish both ends and flare
           both ends  if desired.  Insert a glass wool
           plug at one end, then pack the column uith
           1 g of the! activated AX-21/Celite to form
           a 2 cm long adsorbent bed.  Insert a glass
         ;  wool  plug  on  top of  the bed to  hold the
           adsorbent  in place.

  12.5.2    Pre-rinse   the  column with   five  mL  of
           toluene  followed   by   2  mL   methylene
           chloride:methanol:toluene (15:4:1  v/v),  1
           mL  methylene   chloride:cyclohexane  (1:1
           v/v),  and  five mL  hexane.    If the  flow
           rate  of eluate exceeds  0.5  mL per  min,
           discard the-column.

 12.5.3    When the solvent  is  within  1  rim of  the
           column  packing, apply the sample  ex.tract
         ,  to the  column.   Rinse  the sample container
           twice  with   1  roL  portions of  hexane  and
           apply separately to the  column.   Apply  2
          mL of hexane to complete  the transfer.

 12.5.4   Elute the interfering  compounds with 2 mL
          of   hexane,    2    mL   of   methylene
          chloride:cyclohexane  (1:1  v/v),  and 2 mL
          of   methylene   chloride:methanol:toluene
          (15:4:1 v/v).  Discard the eluate.

 12.5.5    Invert  the  column and  elute  the PCOOs and
          PCOFs with  20 mL of toluene.   If carbon
          particles t.are   present   in   the  eluate,
          filter  through glass fiber filter paper.

 12.5.6    Concentrate  the eluate per Section 11.4 or
          11.5  for further cleanup  or  for injection
          into  the HPLC or GCHS.
      12.6   HPLC (Reference 6)

    12i6.1   Column calibration

 12.6.1.1    Prepare a calibration  standard containing
             the 2,3,7,8-  isomers and/or other isomers
             of interest  at a concentration of approxi-
             mately 500 pg/uL in methylene chloride.

 12.6.1.2   Inject 30 uL  of the calibration solution
             into the HPLC  and  record the  signal  from
             the detector.   Collect the  eluant for re^
            •use.   The elution order will be the tetra-
             through  octa-isomers.

 12.6.1.3    Establish  the  collect time  for  the tetra-
             isomers  and   for  the  other  isomers  of
             interest.    Following  calibration,  flush
             the   injection   system    with    copious
             quantities of  methylene' chloride,  includ-
             ing a minimum of  five  50-uL  injections
             while the detector  is monitored,  to ensure
             that residual  PCDOs and PCOFs are removed
             from the system.

 12.6.1.4   Verify the  calibration with the  calibra-
            tion  solution   after every   20  extracts.
            Calibration is  verified if the recovery of
            the PCDOs  and  PCOFs  from  the calibration
            standard  (Section   12.6.1.1)  is  75-125
            percent   compared   to   the  calibration
            (Section 12.6.1.2).   If  calibration is not
            verified, the system shall be recalibrated
            using the  calibration  solution^   and  the
            previous  20 samples  shall  be re-extracted
            and  cleaned   up  using   the  calibrated
            system.

   12.6.2    Extract cleanup -- HPLC requires  that  the
            coluwi  not  be  overloaded.    The  column
            specified in  this method  is designed  to
            handle  a  maximum of 30 uL of  extract.   If
            the extract cannot be concentrated to  less
            than 30 uL, it  is  split  into  fractions  and
            the fractions are  combined after  elution
            from the column.                  ••

12.6.2.1    Rinse the sides  of the vial twice  with  30
            uL  of methylene chloride and  reduce to 30
            uL  with the blowdown apparatus.

12.6.2.2    Inject the 30  uL extract into  the HPLC.

12.6.2.3   Elute the extract  using the calibration
           data determined  in  12.6.1.    Collect  the
           fraction(s) in a clean 20  mL   concentrator
           tube  containing 5 rnL  of  hexaneracetone
           (1:1 v/v).
                                                                                                           27

-------
12.6.2.4   If an  extract  containing greater than 100
           ng/rnL  of total  PCDO or PCDF  is encoun-
           tered,  a 30 uL methylene  chloride blank
           shall  be run through the  system to check
           for carry-over.

12.6.2.5   Concentrate  the eluate  per  Section 11.5
           for injection into the GCHS.

      13   HRGC/HRHS ANALYSIS

    13.1   Establish  the   operating  conditions given
           in Section 7.1.

    13.2   Add 10 uL of  the  internal standard solu-
           tion (Section  6.12)  to  the sample extract
           immediately prior to  injection  to minimize
           the  possibility of  loss  by evaporation,
           adsorption, or  reaction.   If  an extract  is
           to  be  reanalyzed  and  evaporation  has
        % % occurred,  do   not  add  more   instrument
           internal standard solution.   Rather, bring
           the  extract back  to its  previous  volume
           (e.g.,  19 uL> with pure nonane  only.

    13.3   Inject 1.0  uU  of the concentrated extract-
           containing the  internal  standard solution,
           using   on-colutn  or  splitless   injection.
           Start   the  GC  column  initial   isothermal
           hold   upon   injection.     Start  HS  data
           collection  after the solvent peak elutes.
           Stop data collection  after the  octachloro-
           dioxin and  furan have eluted..   Return the
           column  to   the'  initial   temperature  for
           analysis of  the next extract  or standard.

      14   SYSTEM AND LABORATORY PERFORMANCE

    14.1  -At  the beginning of  each  12-hour shift
           during which analyses are  performed, GCHS
           system  performance   and  calibration  are
           verified for   all  unlabeled  and  labeled
           compounds.   For  these  tests,  analysis  of
           the  CS3 calibration  verification  (VER1
           standard (Section 6.13  and  Table  4) and
           the   isomer  specificity   test  standards
           (Sections 6.16 and Table 5)  shall be used
           to   verify   all   performance   criteria.
           Adjustment   and/or   recalibration    (per
           Section 7)  shall  be performed  until all
           performance criteria are met.   Only after
         .  all   performance  criteria   are met  may
           samples,   blanks,    and   precision  and
           recovery standards be analyzed.
     14.2
HS resolution -- A  static resolving power
of  at  least 10,000  (10  percent  valley
definition) must be demonstrated at appro-
priate  masses  before   any   analysis  is
performed.   Static  resolving  power checks
                                                14.3.1
                                                14.3.2
                                                14.3.3
                                                14.3.4
                                                 14.3.5
                                                         must be  performed at the beginning  and  at
                                                         the end  of each  12-hour  shift  according  to
                                                         procedures  in Section  7.1.2.    Corrective
                                                         actions  must be implemented whenever  the
                                                         resolving   power   does   not   meet   the
                                                         requirement.
                                                         r
                                                  14.3   Calibration verification
Inject   the   VER   standard
procedure in Section 13,.
                                                                                         using   the
The m/z abundance ratios for all PCODs and
PCDFs shall be within t:he limits in Table
3A; otherwise, the mass spectrometer shall
be adjusted until the m/z abundance ratios
fall within the  limits  specified,  and the
verification    test    (Section   14.3.1)
repeated.   If  the adjustment  alters the
resolution of  the mass spectrometeri reso-
lution  shall  be verified  (Section 7.1.2)
prior to repeat of the verification test.

The peaks  representing  each unlabeled and
labeled compound  in the VER standard must
be  present with  a  S/M of  at  least 10;
otherwise, the mass spectrometer shall  be
adjusted   and  the   verification   test
(Section 14.3.1) repeated.

Compute    the    concentration   of   each
unlabeled  compound  by  isotope  dilution
(Section  7.5)  for  those   compounds  that
have  labeled  analogs (Table  1).   Compute
the concentration of  the labeled compounds
by  the .internal  standard method.   These
concentrations are computed  based  on the
averaged  relative  resjxsnse  'and averaged
response  factor  from the calibration data
in Section 7.

For each  compound,  compare the concentra-
tion  with  the   calibration  ; verification
limit  in  Table 7.   If  all compounds meet
the  acceptance criteria,  calibration has
been verified.  If, however,  any compound
fails,  the   measurement  system  is  not
performing properly for that  compound.   In
this  event,  prepare  a  fresh  calibration
standard  or correct  the  problem causing
the  failure   and   repeat   the  resolution
(Section  14.2) ?nd verification (Section
14.3.1) tests, or recalibrate  (Section 7).
                                                              14.4    Retention times and GC resolution
                                                             14.4.1    Retention times
r
r
                                                                                                                  r
  28
r
r
r
r
r
r

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14.4.1.1    Absolute --  The absolute  retention  times
            of  the   13C12-1,2,3,4-TCDO  and     C,,,-
            1,2,3,7,8.9-HxCBD  GCHS  internal  standards
            shall   be  within   ±15  seconds   of   the
            retention times  obtained  during  calibra-
            tion (Section 7.2.4).

14.4.1.2    Relative --  The relative  retention  times
            of unlabeled  and  labeled PCOOs and  PCDFs
            shall  be within the limits given  in  Table
            2-                ..e

  14.4.2 i   GC resolution       ^

14.4.2.1 \   Inject  the   isomer  specificity  standards
            (Section  6.16)   on    their   respective
         '   columns.

14.4.2.2    The valley height  between 2,3,7,8-TCDD and
            the other  tetra-   dioxin  isomers  at  m/z
            319.8965, and between  2,3,7,8-TCOF  and the
            other tetra-  furan  isowers at m/z 303.9016
            shall   not  exceed   25  percent  on  their
            respective columns  (Figure 3).

  14.4.3    If  the  absolute  retention  time  of  any
            compound   is   not   within   the   limits
            specified or  the 2,3,7,8-  isomers  are not
            resolved,   the  GC   is   not  performing
            properly.   In this event,  adjust  the  GC
         ,   and repeat  the verification  test (Section
            14.3.1) or recalibrate  (Section 7).

    14.5    Ongoing precision and accuracy

  14.5.1    Analyze   the   extract  'of   the   diluted
            precision  and  recovery   standard   (PAR)
            (Section  10.3.4  or   10.4.4)   prior   to
            analysis of samples from  the same set.

  14.5.2    Compute the concentration of  each PCDO and
            PCDF   by  isotope   dilution   for   those
            compounds   that   have   labeled    analogs
            (Section 7.5).   Compute  the  concentration
         :   of  each  labeled compound by the internal
            standard method.

  14.5.3    For  each  unlabeled and labeled compound,
           compare the concentration with the limits
            for  ongoing accuracy in  Table 7.   If all
           compounds meet  the acceptance  criteria,
           system   performance  is   acceptable   and
           analysis  of   blanks   and   samples   may
           proceed.  ,, If,  however,  any  individual
           concentration  falls  outside  of  the range
           given,    the    extraction/concentration
           processes are not being performed properly
         •  for that compound.   In this event,  correct
           the  problem,   re-extract  the sample  set
           (Section  10)   and  repeat   the   ongoing
         precision   and   recovery  test   (Section
         14.5).   The concentration  limits  in Table
         7  for  labeled compounds  are based  on  the
         requirement  that   the   recovery  of  each
         labeled  compound be  in  the  range of  25-
         150X.

14.5.4   Add  results  which pass the specifications
         in Section  14.5.3 to initial and  previous
         ongoing  data  for  each  compound  in  each
         matrix.    Update   QC  charts   to   form a ,
         graphic   representation   of   continued
         laboratory performance.   Develop a  state-
         ment of  laboratory  accuracy for each PCDO
         and PCOF  in  each matrix type by calculat-
         ing  the  average percent  recovery  (R)  and
         the standard deviation of percent  recovery
         (SR).  Express  the  accuracy as a  recovery
         interval  from  R -  2S.  to  R  +  2S-.    For
         example,  if  R  = 95X and  SR  *  5X,   the
         accuracy is 85-105X.

    15   QUALITATIVE DETERMINATION

         For  a  gas  chromatographic  peak  to. be
         identified as  a  PCDO or PCDF  (either a
         unlabeled or a  labeled  compound),  it oust
         meet all of the criteria in Sections 15.1-
         15.4.

  15.1    The signals for  the two  exact m/z's being
         monitored (Table 3) must be  present,  and
         must maximize  within *  2  seconds  of  one
         another.

  15.2    The signal-to-noise  ratio (S/N)  of  each of
         the two exact  m/z's must be  greater than
         or  equal  to 2.5  for a sample extract,  and
         greater than  or equal to 10  for a calibra-
         tion  standard   (see  Sections  7.2.3  and
         14.3.3).

  15.3    The ratio of the integrated  ion currents
         of  both the exact m/z's monitored  must be
         within  the limits in Table 3A.

  15.4    The relative retention  time of  the peaks
         representing     a    unlabeled   2,3,7,8-
         substiiuted  PCDO  or PCDF must  be  within
         the  limits   given   in   Table  2.      The
         retention  time  of peaks representing  non-
         2,3, 7,8-substituted  PCOOs  or PCDFs  must  be
         within    the    retention    time   windows
         established in Section 7.3.

 15.5    Confirmatory     analysis     --      Isomer
         specificity for all  of the 2,3.7,8-substi-
         tuted   analytes   cannot   be  attained   by
         analysis  on  the  D8-5 (or  equivalent)  GC
         column  alone.  .The lack of  specificity  is'
                                                29

-------
        of  greatest  concern  for  the  unlabeled
        2,3,7,8-TCDF.   Therefore,  any  sample  in
        which   2,3,7,8-TCDF   is   identified   by
        analysis  on  a  DB-5  (or  equivalent)  GC
        colum  must  have  a confirmatory analysis
        performed on a 08-225, SP-2330,  or equiva-
        lent GC colum.   The operating  conditions
        in  Section  7.1.1   way   be adjusted  for
        analyses on  the second GC  colum, but the
        GCHS  must meet the mass  resolution  and
     >   calibration specifications  in Section 7.

 15.6   If  any gas ehromatographic peak meets the
        identification  criteria   in Sections 15.1,
        15.2,  and  15.4,  but does not meet the  ion
        abundance  ratio criterion  (Section  15.3),
        and is not a  labeled analog, that  sample
        must be analyzed on a second GC  colum, as
        in  Section 15.5 above.   Interferences  co-
      , eluting in  either  of  the  two  m/z's  may
        •cause  the  ion abundance  ratio to fall  out-
        side  of the  limits in  Table  3A.    If  the
        ion abundance ratio of  the peak  fails to
        meet  the criteria  on the second GC colum,
        then  the peak does not represent a POD or
        PCOF.    If the 'peak does meet  all  of  the
        criteria  in  Sections  15.1-15.4  on  the
        second  GC  colum,  then  calculate   the
        concentration  of  that   peak   from   the
         analysis on the second  GC colum, accord-
         ing to the procedures in Section 16.

  15.7    If  any  gas   ehromatographic  peak  that
         represents a  labeled analog does not meet
         all  of  the   identification  criteria  in
         Sections  15.1-15.4  on  the  second  GC
         colum,   then   the  results  may  not  be
         reported    for    regulatory   compliance
         purposes  and  a new aliquot  of the sample
         must be extracted and analyzed.

    16   QUAHTITATIVE DETERMINATION

  16.1    Isotope dilution  --  By  adding  a  known
          amount of  a  labeled  compound  to  every
          sample prior  to extraction,  correction for
          recovery  of the  unlabeled compound can  be
          made   because  the  unlabeled compound and
          its  labeled analog exhibit similar effects
          upon   extraction,  concentration,  and gas
          chronatography.    Relative  response  
-------
16.3
16.4
         The   concentration   of   the   unlabeled
         compound in  the  solid phase of the sample
         is computed using the concentration of the
         compound in  the  extract and the weight of
         the solids (Section 10), as follows:
       Concentration
       in solid (ng/Kg)"
                               
         where,
         V   is the extract volume in mL.
         W  is the sample weight in Kg.

         The   concentration   of   the   unlabeled
         compound  in  the  aqueous  phase  of  the
         sanple is computed using the concentration
         of  the compound  in  the  extract  and the
         volune of water extracted (Section 10.3),
         as follows:
                               ex
                                    Vex>
         Concentration     _   (C
         in aqueous phase  ~-
         (pg/L)                     Vs

         where,
         V   is the extract volune in mL.
       •  V  is the sample volune in liters.

  16.5   If  the  SICP  areas  at  the quantitation
         m/z's for any compound exceed the calibra-
       ,  tion range of the system; a  smaller sample
         aliquot is extracted.

16.5.1  .For aqueous samples containing one percent
         solids  or  less,  dilute  100  mL,   10  mL,
         etc., of  sample  to 1  liter with reagent
         water and extract per Section 11.

16.5.2   For  samples containing  greater  than  one
         percent  solids,   extract  an  amount  of
         sample equal  to 1/10, 1/100, etc., of the
         amount  determined   in   Section  10.1.3.
         Extract per Section 10.4.

16.5.3   If  a  smaller  sample  size  will  not  be
         representative   of   the   entire  sample,
         dilute the  sample extract by  a factor of
         10,   adjust  the   concentration  of  the
         instrument  internal  standard to 100 pg/uL
         in the extract, and analyze an aliquot of
         this  diluted   extract  by  the   internal
         standard method.

  16.6   Results are reported to three significant
         figures  for  the   unlabeled and   labeled
         isomers  found  in all  standards,  blanks,
         and  samples.    For  aqueous  samples,  the.
         units  are  pg/L;  for  samples   containing
         greater  than one' percent  solids  (soils.
         sediments,   filter   cake,  compost),   the
         units are ng/Kg based on  the dry  weight of
         the sample.

16.6.1   Results   for  samples  which   have   been
         diluted  are reported at  the least  dilute
         level at  which the areas at the  quantita-
         tion  m/z's  are  within  the   calibration
         range (Section 16.5).

16.6.2   For  unlabeled  compounds  having a  labeled
         analog,  results are reported at  the  least
         dilute  level  at  which   the  area  at  the
         quantitation m/z  is within the  calibration
         range  (Section   16.5)   and  the   labeled
         compound  recovery  is  within   the  normal
         range for the method (Section 17.4).

16.6.3   Additionally,  the total concentrations  of
         all  isomers  in  an  individual  level   of
         chlorination   (i.e.,   total   TCOD,   total
         PeCDD, etc.) are  reported to three  signi-
         ficant figures  in units of  pg/L,  for both
         dioxins  and furans.   The total  or  ng/Kg
         concentration  in  each  level  of  chlorina-
         tion  is  the sun  of  the concentrations  of
         all  isomers   identified  in  that  level,
         including    any   non-2,3,7,8-substituted
         isomers.

    17   ANALYSIS OF COMPLEX SAMPLES

  17.1   Some samples may  contain  high   levels  (>10
         ng/L;  >1000 ng/Kg)  of  the compounds  of
         interest,   interfering  compounds,   and/or
         polymeric materials.    Some  extracts will
         not  concentrate  to  10  uL   (Section  11);
         others may  overload the  GC  column  and/or
         mass spectrometer.

  17.2   Analyze  a smaller  aliquot  of  the  sample
         (Section  16.4)  when the  extract  will  not
         concentrate  to 20  uL  after all cleanup
         procedures have been exhausted.

  17.3   Recovery  of  labeled   compound, spiking
         standards --  In  most  samples,  recoveries
         of the  labeled compound spiking  standards
         will  be  similar   to  those  .from reagent
         water   or   from   the  alternate   matrix
         (Section 6.6).   If  recovery is outside  of
         the   25-150%  range,   a  diluted   sample
         (Section 16.4) shall  be analyzed.  If  the
         recoveries of  the labeled compound spiking
         standards   in   the  diluted   sample   are
         outside  of  the  limits (per  the  criteria
         above),  then  the  verification   standard
         (Section  14.3)   shall  be   analyzed   and
         calibration verified (Section 14.3.4).   If
         ithe calibration cannot  be verified,  a  new
                                                                                                         31

-------
                                                                                                                             L
          calibration  oust  be  performed   and  the
          original  sample extract  reanalyzed.    If
          the   calibration  is  verified  and   the
          diluted  sample does  not meet  the  limits
          for  labeled compound  recovery,  then  the
          method does  not apply to the sample being
          analyzed   and   the  result   may  not   be
          reported    f r    regulatory   compliance
          purposes.

     18   METHOD PERFOHANCE

          The  performance  specifications   in  this
          method are based on the  analyses of  more
          than  400  samples,   representing   matrices
          from at  least  five industrial  categories.
          These   specifications   will   be  updated
          periodically  as more  data  are  received,
          and each time the procedures  in the  method
        " tare revised.

       REFERENCES

      1   Tondeur,  Yves,  "Method  8290:  Analytical
          Procedures   and  Quality  Assurance   for
          Multimedia   Analysis   of  Polychlorinated
          Dibenzo-p-dioxins  and  Oibenzofurans   by
          High-Resolution  Gas   Chromatography/High-
          Resolution Hass Spectrometry",  USEPA EMSL,
          Las Vegas, Nevada, June 1987.

      2   "Measurement  of  2,3.7,8-Tetrachlorinated
          Dibenzo-p-dioxin (TCOO) and 2,3,7,8-Tetra-
          chlorinated  Dibenzofuran (TCOF)  in  Pulp,
          Sludges, Process  Samples and Waste-waters
          froa Pulp  and Paper Mills",  Wright  State
          University, Dayton, OH 45435, June 1988.

      3   "NCASt Procedures  for  the Preparation  and
          Isomer Specific Analysis  of Pulp  and Paper
          Industry   Samples   for  2,3,7,8-TCDO   and
          2,3,7,8-  TCOF", National Council  of  the
          Paper Industry for Air and Stream Improve-
          ment,  260 Madison  Avenue,  Hew  York,  NY
          10016,  Technical Bulletin  No.  551.  Pre-
          release Copy, Julv 1988.

      4   "Analytical    Procedures    and   Quality
          Assurance  Plan  for  the  Determination  of
          PCDD/PCOF  in Fir.h«,  USEPA,   Environmental
          Research    Laboratory,    6201    Congdon
          Boulevard, Ouluth, HN 55804, April 1988.

      5   Tondeur, Yves,  "Proposed  GC/MS  Methodology
          for  the Analysis  of  PCDDs  and  PCDFs  in
          Special   Analytical    Services    Samples",
          Triangle   Laboratories,    Inc.,     801-10
          Capitola  Or, Research  Triangle  Park,  NC
          27713,  January 1988;  updated by  personal
          communication September 1988.
 6   Lamparski,   L.L.,   and   Hestrick,   T.J.,
     "Determination  of  Tetra-,  Hexa-,  Hepta-,
     and Octachlorodibenzo-p«dioxin  Isomers in
     Particulate Samples at  Parts per Trillion
     Levels",  Analytical  Chemistry.  52:  2045-
     2054, 1980.

 7   Lamparski,   L.L.,   and   Nestrick,   T.J.,
     "Novel    Extraction    Device .   for    the
     Determination  of  Chlorinated  Dibenzo-p-
     dioxins (PCDDs)  and  Dibenzofurans (PCDFs)
     in     Matrices     Containing     Water",
     Chemosphere. 19:27-31, 1989.  !

 8   Patterson,  D.G.,  et.   al.  "Control   of
     Interferences  in  the  Analysis  of  Human
     Adipose    Tissue    for    2,3,7,8-Tetra-
     chlorodibenzo-p-dioxin".     Environmental
     Toxicological Chemistry. 5: 355-360,  1986.

 9   Stanley,  John  S.,  and  Sack,  Thomas  M.,
     "Protocol  for  the Analysis  ;of  2,3,7,8-
     Tetrachlorodibenzo-p-dioxin    by    High-
     Resolution     Cas     Chromatography/High-
     Resolution Mass Spectrometry", USEPA EMSL,
     Las Vegas, Nevada 89114, EPA 600/4-86-004,
     January 1986.

10   "Working  with  Carcinogens",  OHEW,   PHS,
     CDC,  NIOSH,  Publication  77-206,  August
     1977.

11   "OSHA Safety and Health Standards, General
     Industry" OSHA  2206,  29 CFR 1910, January
     1976.

12   "Safety      in     Academic     Chemistry
     Laboratories",  ACS  Committee  on Chemical
     Safety, 1979.

13   "Standard Methods  for  the  Examination of
     Water  and Wastewater",  16th  edition  and
     later  revisions, American Public  Health
     Association,    1015    15th    St,    N.U.,
     Washington,  DC  20005,  46:  Section  108
     (Safety),  1985.

14   "Method 613 --  2,3,7,8-Tetrachlorodibenzo-
     p-dioxin",  40  CFR  136  (49  FR  43234),
     October 26, 1984, Section 4.1.

15   Provost,    L.P.,   and    Elder,    R.S.,
     "Interpretation of Percent Recovery Data",
     American Laboratory.  15: 56-83, 1983.

16   "Handbook of Analytical Quality Control in
     Water and Wastewater  Laboratories",  USEPA
     EMSL, Cincinnati, OH  45268, EPA-600/4-79-
     019, March 1979.
r
r
t
r
32
c
r
L
L
r
r

-------
17   "Standard  Practice  for  Sampling  Water",
     ASTM Annual  Book of Standards, ASTM, 1916
     Race Street,  Philadelphia,  PA 19103-1187,
     1980.

18   "Methods   330.4   and   330.5   for  Total
    ', Residual  Chlorine", USEPA,  EMSl,  Cincin-
     nati,  OH  45268,  EPA  600/4-70-020,  March
     1979.

19   Barnes, Donald G.,  Kutz.  Frederick U., and
     Baltimore,  David P.,  "Update  of Toxicity
     Equivalency  Factors CTEFs)  for Estimating
     Risks   Associated   with   Exposures   to
     Mixtures  of Chlorinated  Dibenzo-p-Oioxins
     and   Dibenzofurans   (CDDs/CDFs)",   Risk
     Assessment  Forua,  USEPA, Washington,  DC
     20460, February 1989.
                                                                                                      33

-------
                                                    Table 1
         POLYCHLORINATED DIBENZOOIOXINS AND FURANS DETERMINED  BY  ISOTOPE DILUTION AND  INTERNAL  STANDARD
               HIGH RESOLUTION GAS CHROMATOGRAPHY (HRGO/HIGH  RESOLUTION MASS  SPECTROMETRY  (HRMS)
PCODs/PCOFs (1)
Isomer/Congener
2,3,7,8-TCOD
*
Total-TCDO
2,3 7,8-TCDF
Totul-TCOF
1,2,3,7,8-PeCDD
Total-PeCOO
1,2,3,7,8-PeCOF
2,3,4,7,8-PeCOF
Total-PeCDF
1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCOD
1,2,3/7,8,9-HxCOD
Total-Hx'COD
1,2,3,4,7,8-HxCDF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9- HxCDF
2,3,4,6,7,8-HxCOF
Total -HxCDF
1,2,3,4,6,7,8-HpCOD
Total-HpCOD
1 ? ^ L ft 7 8-HnCDF
I ,£,.?, *»,Q,/ ,o n|A»ur
1 ? ^ L 7 8 9-rioCOF
•i&t^t^t ' t°i* npuur
. Total-HpCDF
OCOD
OCOF
(1) Polychlorinated dioxins

CAS Registry
1746-01-6
*
41903-57-5
51207-31-9
55722-27-5
40321-76-4
36088-22-9
57117-41-6
57117-31-4
30402-15-4
39227-28-6
57653-85-7
19408-74-3
34465-4608
70648-26-9
57117-44-9
72918-21-9
60851-34-5
55684-94:1
35822-46-9
37871-00-4
67562-39-4
55673-89-7
38998r75-3
3268-87-9
39001-02-0
and furans
TCDD " Tetrachlorodibenzo-p-dioxin
PeCOD 3 Pentachlorodibenzo-p-dioxin
HxCDD * Hexachlorodibenzo-p-dioxin
HpCOD => Heptachlorodibenzo-p-dioxin
OCDD a Octachlorodibenzo-p-dioxin

Labeled Analog
"c12-2,3,7.8-TCDD
37Cl4-2,3,7,8-TCOD

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

13C12-1,2,3,7,8-PeCOD

13C12-1,2,3,7,8-PeCDF
13Cl2-2,3,4,7,8-PeCDF
.
•]3C12-1,2.3,4,7,8;HXCDD
13C12- 1,2,3,6,7,8-HxCOD
13C12- 1,2,3, 7,8,9- HXCODC2)

13C12-1,2,3,4,7,8-HxCOF
13Cl2-1,2,3,6,7,8-HxCOF
13C12-1, 2,3, 7,8,9- HxCOF
13C12-2,3,4,6,7,8-HxCDF


13C12-1,2,3,4,6,7,8-HpCDD



12
13C12-OCOD '
none

TCDF = Tetraehlor'odibenzofuran
PeCDF = Pentachlorodibenzofuran
HxCDF = Hexachlorodibenzofuran
HpCDF 3 Heptachlorodibenzofuran
OCDF = Octaehlorodibenzofurari

CAS Registry
76523-40-5
85508-50-5

89059-46-1

109719-79-1

109719-77-9
116843-02-8

109719-80-4
109719-81-5
109719-82-6

114423-98-2
116843-03-9
116843-04-0
116843-05-1

109719-83-7

109719-84-8
109719-94-0

114423-97-1







<2>  Labeled analog is  used as an internal  standard and therefore is not  used for quantitation of  the  native
     cotrpound.
                                                                                                                                r
                                                                                                                                r
                                                                                                                                r
 r
 r
 r
 r
 r
r
 r
r
r
                                                                                                                               •4,
 34

-------
                                                    Table 2
                            RETENTION TIMES AND MINIMUM LEVELS FOR PCDDs AND  PCDFs
                                                                                          Minimum Level (1)
Compound .
Compounds using C12* 1,2,3,
Native Compounds
2,3,7,8-TCDF
2,3,7,8-TCDD
1,2,3,7,8-PeCDF
2;3,4,7,8-PeCDF
1,2,3,7,8-PeCDO
Labeled Compounds
l3C1.-2,3,7,8-TCDF
IT 12
13C12-1,2,3,4-TCDD
13C1,-2,3,7,S-TCDD
T7 12
•i/Cl4-2,3,7,8-TCDD
13C12-1,2,3,7,8-PeCOF
13C12-2,3,4,7,8-PeCDF
13C12-1,2,3,7,8-PeCDD
Compounds using C12~ 1,2,3,
Native Compounds
1,2,3,4,7,8-HxCOF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3, 4,7,8- HXCDD
1,2,3,6,7,8-HxCDD
1,2,3, 7,8, 9- HxCDD
1,2,3,4,6.7,8-HpCDF
1,2,3,4,6,7,8-HpCDD
1,2,3,4,7,8,9-HpCDF
OCDD.
'. OCDF
Labeled Compounds
13C12-1,2,3,4,7,8-HxCDF
13C12- 1,2,3,6,7,8-HXCDF
13Cl2-1,2,3,7,8,9-HxCDF
13C12-2,3,4,6,7,8-HxCDF
13C12-1,2*3,4,7,8-HxCOD
13C12- 1 ,2, 3,6,7,8-HxCDD
13C,_- 1,2,3, 7,8, 9-HxCOD
1*1 1 £
^C12-1,2,3.4,6,7.8-HpCDF
"c12-1,2,3,4,6,7.8-HpCOO
13C12-1, 2,3,4,7 8,9-HpCDF
13C12.-OCDD
Retention
Time
Reference
4-TCDD as internal standard

13C12-2,3,7,8-TCDF
13C12-2,3,7,8-TCDD
13C12-1,2,3,7,8-PeCDF
13C12-2,3,4,7,8-PeCDF
13C12-1,2,3,7,8-PeCDD

13C12-1,2,3,4-TCDD
3C12-1,2,3,4-TCDD
J3C12-1. 2.3.4- TCOD
J3C12- 1,2.3, 4-TCDD
]3C12-1,2,3,4-TCDO
"C12-1, 2,3,4- TCDD
13C12-1,2,3,4-TCDD
7, 8, 9- HxCDD as internal standard

13C12-1,2,3,4,7,8-HXCDF
13C12-1,2,3,6,7,8-HxCDF
13C12-1,2,3,7,8,9-HxCDF
3C12-2,3,4,6,7,8-HxCOF
13C12-1, 2,3,4, 7,8-HxCDD
13C,,-1,2,3,6,7,8-HxCDD
n ib
•3C12-1,2,3,6,7,8-HxCDD
]3C12-1,2,3,4,6,7,8-HpCDF
]3C12-1,2,3,4.6,7,8-HpCDD
13C12-1,2,3,4,7,8,9-HpCOF
3C12-OCDD
I3c12-ocoo

13C12-1.2,3,7,8,9-HxCOD
13C12- 1,2,3,7,8,9- HxCDD
13C12-1, 2,3,7.8,9- HxCOD
13c'12-1, 2,3,7,8,9- HxCOO
13C12- i2,3,7,8,9-HxCDD
13C,_- ,2,3,7.8,9-HxCOD

C12- ,2,3,7,8,9-HxCDD,
13C12- ,2,3,7,8,9-HxCDD
13C12- ,2,3.7.8,9-HxCDO
13C12- ,2,3,7,8,9-HxCDO
13C12-1, 2,3,7,8,9- HxCDD
Relative
Retention
Time


0.993 -
0.993 -
0.918 -
0.999 •
0.987 -

0.931 -
1.000 •
0.993 -
1.002 -
1.091 -
1.123 -
1.134 -


0.936 -
0.973 -
0.937 -
0.999 -
0.999. -
0-.992 -
0.936 -
0.930 -
0.986 -
0.896 -
0.996 -
0.995 -

0.947 •
0.940 •
0.993 -
0.971 -
0.974 -
0.975 -
1.000 -
0.953 -
1.023 -
1.024 -
1.050 -


1.009
1.009
1.076
1.001
1.016

0.994
1.000
1.036
1.013
1.371
1.408
1.428


1.015
1.025
1.068
1.001
1.001
1.009
1.016
1.022
1.016
1.079
1.005
1.013

0.992
1.006
1.017
1.000
1.002
1.006
1.000
1.172
1.125
1.148
1.275
Water
pg/L
ppq


10
10
so
so
50










50
50
50
50
50
50
50
50
50
50
100
100












Solid
ng/kg
ppt


1
1
5
5
5










5
5
5
5
5
5
5
5
5
5
10
10












Extract
pg/uL
PPO


0.5
. 0.5
2.5
2.5
2.5










2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
5.0
5.0









'


(1)  Level at which the analytical system will give acceptable SICP and  calibration.
                                                                                                           35

-------
                                      Table 3
DESCRIPTORS, MASSES, M/Z TYPES, AND ELEMENTAL COMPOSITIONS OF THE CODs AND  CDFs (1)
Descriptor Accurate
Number 	 m/z (2)
" 1 292.9825
303.9016
305.8987
315.9419
317.9389
319.8965
321.8936
327.8847
"*• 330.9792
»
331.9368
333.9339
375.8364
2 339.8597
341.8567
351.9000
353.8970
354.9792
355.8546
357.8516
367.8949
369.8919
409.7974
3 373.8208
375.8178
383.8639
385.8610
389.8157
391.8127
392.9760
401.8559
403.8529
430.9729
445.7555
36
m/z
Type Elemental Composition
Lock C7 F11
" C12H435CI4°
M+2 C12 H4 35C1337CI 0
% «4 ^ °
% H, 35d3 37Cl 0
H . C12 H4 35cl4 °2
M+2 C,2 H4 35C13 37Cl 02
H C12H437C1402
QC C7 Fn

"l^^Cl^ClO,
M+2 C12 H4 35C15 37Cl 0
M+2 - C12 H3 35C14 37Cl 0
H+4 C12 H3 35C13 37C12 0
H+2 13C12 H, 35C14 37Cl 0
M+4 " 13C12 H3 35C13 37C12 0
Lock C0 F13 .
H+2 C12 H3 35Ct4 37Cl 02
H+4 C12 H3 35d3 37C12 0,
H+2 13C12 H3 35C14 37Cl 02
H+4 . 13C12 H, 35C13 37C12 O,
"« '12 "3 35'16 37CI °
H+2 C12 H2 3SClg 37Cl 0
H+4 C12 H2 35C14 3 C12 0
H 13C12 H2 35C16 0
:H+2 13c12 H2 35ci5 37ci o
H+2 C,, H2 35C15 37Cl O,
H+4 C12 H2 35C14 37C12 02
.Lock C9 F15
H+2 13C12 H2 35C15 37Cl 02
H+4 13C12 H, 35C14 37C12 O,
QC C, F13
H+4 C12 H2 35C16 37C12 0

Compound
PFK
TCDF
TCDF
TCDF<4)
TCDF<4)
TCDD
TCDD
TCDD (5)
PFK
TCDDC4)
TCDDC4)
HxCDPE
PeCOF
PeCDF
PeCDF(4)
PeCDF<4)
PFK
PeCOD
PeCOD
PeCDOriJ
PeCDD(4)
HpCOPE
HxCDF
HxCDF
HXCDFC4)
HxCOF(4)
HxCBD
HxCDD
PFK
HxCDD<4:>
HXCDOC4)
PFK
OCDPE
H^^BJJH
Primary
: m/z?

! Yes

Yes

Yes



Yes


Yes

Yes


Yes

Yes


; Yes

Yes

Yes
' |

Yes



••^H

-------
                                                Table 3  (continued)
                DESCRIPTORS,  MASSES,  M/Z  TYPES,  AND ELEMENTAL COMPOSITIONS OF THE CODs AND CDFs (1)
Descriptor Accurate m/z
Number ' m/z (2) Type



















(1)
(2)


(3)






4 " 407.7818 M+2
409.7789 M+4
417.8253 H
419.8220 M+2
423.7766 M+2
425.7737 M+4
430.9729 Lock
. 435.8169 . M+2
437.8140 M+4
479.7165 M+4
5 441.7428 M+2
442.9728 Lock

443.7399 M+4
457.7377 M+2
459.7348 M+4
469.7779 M+2
471.7750 M+4
513.6775 M+4 '
From Reference 5
Nuclidic masses used:
H = 1.007825 C 3 12.00000
0 a 15.994915 35Cl = 34.968853
Compound abbreviations:
Chlorinated dibenzo*p-dioxins
TCDD 3 Tetrachlorodibenzo-p-dioxin
PeCDD = Pentachlorodibenzo-p-dioxin
HxCDD = Hexachlorodibenzo-p-dioxin
HpCDD 3 Heptachlorodibenzo-p-dioxin
OCDD = Octachlorodibenzo-p-dioxin
Elemental Composition
C12 H 35C16 37Cl 0
C12 H 35C15 37C12 0
13C12 H 35C17 0
13C12 H 35C16 37Cl 0
C12 H 35C16 37Cl 02
C12 H 35C15 37C12 02
C9F17
13C,_ H 35Cl, 37Cl 0-
\£. O c.
13. „ 35 . 37 .
C12 H C15 C12 °2
C12 H 35C17 37C12 0
c12 35d7 37ci o
C,n F,_
10 17
C 35Cl 37Cl 0
C12 C16 C12 °
C 35Cl 37Cl 0
C12 C17 Cl °2
C 35Cl 37Cl 0
C12 C16 C12 °2
13C12 35C17 37Cl 02
13 35 37.
C12 C16 C12 °2
C 35Cl '37Cl 0
C12 C18 C12 °


13C = 13.003355
37Cl = 36.965903

Compound
(3)
HpCOF
HpCDF
HpCDF(4)
HpCDF(4)
HpCDO
HpCDD
PFK
HpCDO(4)
HpCDD(4)
NCDPE
OCDF
PFK

OCDF
OCDD
OCDD
OCDD (4)
OCDD(4)
DCOPE


F = 18.9984


Primary
m/z?
Yes

Yes

Yes


Yes


Yes



Yes

Yes







Chlorinated diphenyl ethers
HxCOPE »
HpCDPE =
OCDPE =
NCDPE =
DCOPE =
Hexachlorodiphenyl ether
Heptachlorodiphenyl ether
Octachlorodiphenyl ether
Nonachlorodiphenyl ether
Decachlorodiphenyl ether

*



     Chlorinated dibenzofurans
        TCDF     =     Tetrachlorodibenzofuran
        PeCDF    =     Pentachlorodibenzofuran
        HxCDF    a     Hexachlorodibenzofuran
        HpCDF    =     Heptachlorodibenzofuran
Lock mass and QC compound
   PFK      =    Perfluorokerosene
<4)  Labeled compound
                               37
<5)  There is only one m/z for   Cl,-2,3,7,8-TCDD (cleanup standard).
                                                                                                            37

-------
                        Table 3A
   THEORETICAL  ION ABUNDANCE RATIOS AND CONTROL LIMITS

 No. of         m/z's
Chlorine       Forming   Theoretical  Control Limits(l)
 Atoms	Ratio	Ratio	Lower   Upper

 •4  <2>        H/H+2       0.77        0.65    0.89
   5            M+2/H+4      1.55        1.32    1.78
   6            H+2/H+4      1.24        1.05    1.43
   6  (3)        H/H+2       0.51        0.43    0.59
   7    •        H+2/H+4      1.05        0.88    1.20
   7  (4)        H/H+2       0.44        0.37    0.51
   8            H+2/H+4      0.89        0.76    1.02

(1)  Represent  + 15X windows around  the theoretical  ion
                                                                                                                             L*
                                                                                                                             r
                                                                                                                             r
                                                                                                                             r
                                                                                                                             c
     abundance ratios.                                                                             :                           I"
(2)  Does not apply to   Cl,-2,3,7,8-TCDD  (cleanup
     standard).
(3)  Used for 13C-HxCDF  only.
(4) .Used for 13C-HpCDF  only.
 38
                                                                                                                             r
                                                                                                                             L
                                                                                                                             C
                                                                                                                             c

-------
                                                     Table 4
                 CONCENTRATIONS OF SOLUTIONS CONTAINING LABELED  AND  UNUBELED  PCDOS AND PCDFS  ••
                                           STOCX AND SPUING SOLUTIONS
                                   Labeled
           Compound
                                    Stock
                                 Solution (1)
                                    (ng/iBL)
   Labeled
  Coinpound
   Spiking
Solution <2)
   (ng/iK.)
    PAR
   Stock
Solution (3)
   (ng/irt.)
   Cleanup
  Standard
   Spiking
Solution <4)
   
  Internal
  Standard
  Spiking
Solution (5)
  (ng/mt)
Native CDOs and CDFs
             2,3,7,8-TCDO
             2.3.7.8-TC8F
          1,2,3,7.8-PeCOO
          1,2.3.7,8-PeCOF
          2,3.4,7,8-PeCDF
        1,2.3,4,7,8-HxCDD
        1.2,3,6,7.8-HxCDD
        1.2.3,7,8,9-HxCOO
        1,2,3.4,7,8-HxCDF
        1,2,3,6,7,8-HxCDF
        1,2,3.7,8,9-HxCDF
        2,3,4,6,7,8-HxCDF
      1,2.3.4,6,7,8-HpCOO
      1,.2,3.4,6,7,8-HpCDF
      1,2,3,4.7,8,9-HpCOF
                      OCDF
 Labeled  CDOs  and CDFs
       13C12-2.3,7,8-TCOO
       13C12-2,3,7,8-TCDF
     13C12-1.2,3.7.S-PeCDO
.    13C12-1,2,3,7,8-PeCOF
-    13C12-2,3,4,7,3-PeCDF
   13C12-1,2,3,4.7,8-HXCDO
   ]3C12-1,2,3,6,7,8-HxCOO
   13C12-1,2,3,4.7.8-HxCDF
   13C12-1,2,3,6,7,3-HxCOF
   13C12-1,2,3,7,8,9-HxCOF
   13C12-2,3,4.6,7,8-HxCDF
 ]3C12-1,2,3,4,6,7,8-HpCDO
 ]3C12-1,2,3,4,6,7,8-HpCDF
 13C12-1,2,3,4,7,8,9-HpCDF
                13C12-OCDD
 Cleanup  Standard
       37Cl4-2,3,7.8-TCDD
 Internal Standards
       13C12-1,2,3,4-TO»
   13C,2-1.2,3,7.8,9-HXCDO
                                     100
                                     100
                                     100
                                     100
                                     100
                                     100
                                     100
                                     100
                                     100
                                     100
                                     100
                                     100
                                     100
                                     100
                                     200
                      40
                      40
                     200
                     200
                     200
                     200
                     200
                     200
                     200
                     200
                     200
                     200
                     200
                     200
                     200
                     400
                     400
                                                                                      0.8
                                                                                                     200
                                                                                                     200
(1)  Section 6.10 - prepared in nonane and diluted to prepare spiking solution.
(2)  Section 10.3.2 - prepared fron stock solution daily.
(3)  Precision and Recovery (PAR) standard.  Section 6.14 - prepared in nonane and diluted to prepare spiking
     solution in Section 10.3.4.
(4)  Section 6.11 - prepared in nonane.
(S)  Section 6.12 - prepared in nonane.
                                                                                                            39

-------
                                             Table 4 (continued)
                CONCENTRATIONS OF  SOLUTIONS CONTAINING LABELED AND UNLABELED PCDDS AND PCDFS
                                    CALIBRATION AND VERIFICATION SOLUTIONS
	 Compound 	 .
Native CDDs and CDFs
2,3,7,8-TCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDD
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,4,7,8-HxCDD
1,2,3, 6,7, 8-HxCDD
1, 2,3,7,8,9- HxCDD
1,2.3,4,7,8-HxCDF
1,2,3,6,7,8-HxCDF
"1,2,3',7,8,9-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDD
1,2,3,4,6,7,8-HpCOF
1,2,3,4,7,8,9-HpCDF
OCDD
OCOF
Labeled CODs and CDFs
13C12-2,3,7,8-TCDD
13C12-2,3.7,8-TCOF
]3C12-1,2,3,7,S-PeCDD
]3C12-1,2,3,7.8-PeCDF
13C12-2,3,4,7,8-PeCOF
13C12-1,2,3,4,7,8-HxCDD
13C12-1,2,3,6.7,8-HXCOD
13C12-1,2,3,4.7,8-HxCDF
13C12-1,2,3,6,7,8-HXCDF
13C12-1,2,3,7,8,9-HXCOF
13C12-2,3,4,6,7,8-HxCDF
13C12-1,2,3r4,6,7,8-HpCOD
]3C12-1,2,3,4,6,7,a-HpCDF
13C12-1,2,3,4,7.8,9-HpCDF
13C12-OCDD
Cleanup Standard
37Cl4-2,3,7,8-TCOD
Internal Standards
13C12-1,2,3,4-TCDD
13C12-1,2,3,7,8,9-HxCDD
CS1

0.5
0,5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
5.0
5.0

100
100
100
100
100
100
100
100
100
100
100
100
100
100
200

0.5

100
100
CS2
(ng/mL)

2
2
10
10
10
10
10
10
10
10
10
10
10
10
10
20
20

100
100
100
100
100
100
100
100
100
100
100
100
100
100
200

2

100
100
VEIU6)
CS3
(ng/mL)

10
10
50
50
50
50
50
50
50,
50
50
50
50
50
50
100
100

100
100
100
100
100
100
100
100
100
100
100
100
100
100
200

10

100
100
CS4
(ng/mL)

40
40
200
200
200
200
200
200
200
200
200
200
200
200
200
400
400

100
100
100
100
100
100
100
100
in
100
100
100
100
100
200

40

100
100
CSS
(ng/mL)

200
200
1000
1000
1000
1000
1000
1000
1000
: 1000
1000
1000
1000
: 1000
1000
2000
2000

: 100
100
', 100
100
100
100
100
: 100
100
100
100
100
100
100
200

200

100
; 100
L<
[
                                                                                                                            r-
(6)  Section 14.3 -  calibration verification (VER) solution.
 40

-------
                         Table 5
 GC RETENTION TIME WINDOW DEFINING STANDARD MIXTURES AND
        ISOMER SPECIFICITY TEST STANDARD MIXTURES

DB-5 Column GC Retention Time Window Defining Standard
(Section 6.15)
Congener         First  Eluted	Last Eluted
TCDF
TCDD
PeCDF
PeCDO
HxCDF
HXCOD
HpCDF
HpCOD
1,3,6,8-
1,3,6,8-
1,3,4,6,8-
1,2,4,7,9-
1,2,3,4,6,8-
1,2,4,6,7,9-
1,2,3,4,6,7,8-
1,2,3,4,6,7,9-
1.2.8,9-
1.2,8,9-
1,2,3,8,9-
1.2,3,8,9-
1,2,3,4,8,9-
1,2,3,4,6,7-
1,2,3,4.7,8,9-
1,2,3,4,6,7,8-
DB-5 TCDD Isomer Specificity Test Standard
(Section 6.16.1)
                  1,2,3,4-TCDD          1,2,3,7-TCDD
                  1,2,7,8-TCDD          1,2,3,8-TCDD
                  1,4,7,8-TCDD          2,3,7,8-TCDD

DB-225 Column TCDF Isomer Specificity Test Standard
(Section 6.16.2)
                  2,3,4,7-TCOF
                  2,3,7,8-TCDF
                  1,2,3,9-TCDF
                                                                                                           41

-------
REFERENCE
2,3,7,8-TCDD
2.3.7,8- TCOF
1,2,3,7,8-PeCDD
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCOD
1,2,3,7,8,9-HxCDD
1,2,3,4,7,8-HxCDF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCOF
2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDD
1,2.3,4,6,7,8-HpCOF
1,2,3,4,7,8,9-HpCDF
OCDD ,
OCDF

Table
COMPOUNDS FOR QUAHTITATION OF
Reference Compound
13C12-2,3,7.8-TCDD
13C12-2.3,7.8-TCDF
13C112-1,2,3.7.S-S*eCDF
13C12-2,3,4,7.8-feCDF
13C12-1. 2.3,4,7,8- 1XCOD
13C12-1,2,3,6,7,8-HXCDD
(1)
13C12-1,2,3f6f7,8-HxCDF
13C12-2,3,4,6,7,S-HXCDF
l3Cl2-1,2.3,4,6,7,8-HpCDD
13Cl2-1,2,3.4,6,7,8-HpCDF
13C12-1,2.3,4,7,8,9-HpCDF
13C,,-OCDD
11 '*
"C^-OCDD

6
NATIVE AND LABELED PCDDS AND
Labeled PCDDs and PCDFs
13C12-2,3,7.8-TCOD
13C12-2,3,7.8-TCOF
13C12-1.2,3.7.8-PeCDD
13Cl2:1.2,3,7,8-PeCDF
13C12-2,3,4,7.8-PeCDF
13Cl2-1.2.3f4.7,8-HxCDD
13C12-1.2.3,6.7.8-HXCOD
13C12-1.2,3,7,8,9-HXCDD
13C12-1,2,3,4,7,8-HXCDF
13C12-1, 2,3,6. 7,8;HXCDF
13C12-1,2,3,7.a,9-HxCDF
13C12'2,3,4,6,7,8-HXCDF
13C12-1.2,3,4,6,7.8-HpCDO
13C12-1f2,3,4,6,7.8-HpCDF
13C12-1,2.3,4,7,8,9-HpCDF
13C12.OCDD
37Cl4-2.3,7.8-TCDD
	 *„ »k_ ^^r -1 ?_3.4
PCDFS
Reference Compound
13C12-1,2,3,4-TCDD
13C12-1,2,3,4-TCDO
13C12-1,2,3,4-TCDD
13C12-1,2,3,4-TCDD
13C12-1,2,3f4-TCDD
13C12-1,2,3,7,8,9-HXCDD
13C12-1,2,3,7,8,9-HXCDD
13C12-1.2.3.7J,8,9-HXCOD
13C12-1,2,3,7,8,9-HXCDD
13Cl2-l,2.3,7,8f9-HxCDD
13Cl2-1,2,3,7,a,9-HxCDD
13C12-1,2,3,7,8,9-HXCDD
13C12-1,2,3,7,8,9-HxCDD
13C12-1,2,3,7r8,9-HxCOD
13C12-1,2,3,7,a,9-HxCDD
13C12-1,2,3.7,8,9-HxCDD
13C12-1;2,3,4-TCDD
.7.8-HxCDD and C---
(1)  1,2,3,7,8,9-HxCDD  is
     1,2,3,6,7,8-HxCDD.
quantified using the average  responses for the   Cl2
                                                                                                                            r
                                                                                                                            r
                                                                                                                            r
                                                                                                                            c
   42

-------



THIFID REVISION



>


TO METHOD

. i :


1613 PERFORMANCE S
Tafate 7 :
; ACCEPTASCE CRnSSZA. FOK PERFORHMiC£ tESTS
: T«t
Cooe. CU G .
Compound Cng/flt) Cng/itJ-
2,3,7.8-rGDD
2.3,7,8-rCDF
1.2r3,7,S-PeCDO
1,2.3.7,8-PeCDF
2,3.4.7,8-PeOJF
1 ,2,3,4,7,8-HxCDO
1, 2,3.6. r,S-Hxa»
1. 2^5,7,8. 9-H*n»
1,2,3,4,7.3-axCOF .
1 ,2^.6.7.8-HXCDF
1,2.3,7,8,9-lfcCJF
1 -2 .? •£ A T X.UrSTTln
; 10 1.1
10 0.9
,50 3.6
=50 3.4
ISO 4.2;
-50 6.7
50 3.9
;SO 7.0
SO S-5 '
ISO 3.0
-50 2.9
SO 4.2
'cn T0 r
IF* C2> :
* OFR{23
8.3 - 11.8 6,9 - 13.
8.4 - 13.2 ' 6.9 - 15.
41.4 - 56.8 35.7 - £S.
43.0 - SS.O 35,7 - 69.
«2-7 - 61.5 34.3 - 72.
40.8 - 67.1 36-S - 70.
42.9 - S7.S 42^ - F 100
^12-l»2,3,7,8-PeCDO 100
^ci2-t.2,3,7.8-PeCOF 100
"Cl2-2.3;4,r,8-Pe£3»F 100
*fCl2-!.2,3;4,7.8-Hxa» 100
ci2-i,2A6/7,a-atfeo 100
'HC12-1,2,3,4,7^-H>S»F 100
Ci2-1,2;3A7,8-aF 100
Ct2"1«2^t^7J8-9*fi3rf!DF 100
13 •• ••
^ Ct2-2A4,6,7.8-HtfOF 100
Ci2rl^^,4,6,7,S-8paiO 100
^2-l,2,3,4,6,7,8^pO»F 100
^2"1'2>3**'7*?'9"HPa)£: 1°°
C12~OCDO 200
	 	 L 	
. 1S.»
20-S
3Z.1 •
as •
25,1
24^-,
31.4
19.9
15.1 .
1&.X
**°
17.3
20.9
23^
22.9
4SJ5
-
CU All specificariaos are sivw as eotKantrsiHons
C23 s = standard devfatfcn of tiJe cofa-~^— «^' 	 ~-
coopountfe in IPR and OPS eliquits.
8.2^ and 14^,3J.
Re^sed 2/28/92

•*a<^% «x^.iu«t~ A
ere based on

t i .
25.0 -
-25JJ -
25.0 -
25.0 •
25-0 -
• 2510 -
' 25.0 -
: 25-0 -
25.0 -
9? it
O.w -
25.0 -
25.0 -
25.0 -
25.0 -
5Q.O -
Z.3 -
in tie f Ire
= av«r»s« <
requTresMnt


150.0:-
: 150.0;
! 150.0;
iso.o:
i 150,0-'
; iso.o.
150-0 ;
150.0 i
1SO.O;
• _
150.0
; iso.o •
; 150.0:
150.0 ;
150.0 i
300.0!
15.0 :
fl extract or :
aocentrarioa,
s far Labeled
i

25,0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25JJ -
2S.« -
25.0 -
_^
2SJ3 _ -
25.0 -
2SJJ -
2S.O -
2S.O -
50,0 -
3LS.-
150.6
isp,o
150.0
150.0
• 150.0
: isb-d
iso.o;
150.0:
' 750.0
: " J
. 159.0
150-0
150.01
150.01
150.oi
'30Q.oi
I5_oi
standcfX* SoLut.ioiw
Concentration iis
ecepouad recovery


. i
:
: . :
82.0 -
71.0 -
62.4 -
6&.7 r
SS.O -
. 81,8 -
79JS -
71.4 -
65.3 -:

75.4 -
73^5 -
63.7 -'
S5.1 -
60.O -
94^ -
7.1 -:.
118.0
129.0
137.6
133,3
12S.O
118.2
120.7
IZS^
134.7

124^6
126,6
T45^
113.5
140.0
«2.S
12.9
ifts for labeled
erf 25-150K CSeccfons



, • .*
                                                                                                 -  59,2
 40^  -
 79.7  -  125,«
'71^  -  -B9.7

-------
                                                                                                                         L
                                                  Table S
                           SAMPLE PHASE AND QUANTITY EXTRACTED FOR VARIOUS MATRICES
Sample Matrix (1)
. SINGLE PHASE
Aqueous
Example
Drinking water
Percent
Solids
<1
Phase
(2)
Quantity
Extracted
1000 mL
      Solid



      Organic



   MULTIPHASE
      Liquid/Solid

       ..Aqueous/solid
        Organic/solid


      Liquid/Liquid
        Aqueous/organic
        Aqueous/organi c/
        solid
                         Groundwater
                         Treated wasteuater

                         Dry soil
                         Compost
                         Ash

                         Waste solvent
                         Waste oil
                         Organic polymer
                          Wet  soil
                          Untreated effluent
                          Digested municipal sludge
                          Filter cake
                          Paper pulp
                          Tissue
                          Industrial sludge
                          Oily waste
                          In-process effluent
                          Untreated effluent
                          Drum waste

                          Untreated effluent
                          Drum waste
                                                         1-30
                                                             1-100
                                                                               Solid
Organic
                                                                           Solid
                                                                                Both
                                                                          Organic
Organic
& solid
10 g


10 g





10 g





10 g



10 g



10 g
                                            i_
(1)
(2)
The exact matrix may be vague for sons  samples.  In general, when the CDDft andi CDFs are |"'
multiphasa system in which one of the phases is water, they will be preferentially dispersed in or adsorbed
on the alternate phase,  because of their low solubility in water.
Aqueous samples are filtered  after spiking with labeled analogs.  The filtrate and  the material trapped on
the filter are extracted separately, and then the extracts are combined for cleanup  and analysis.
 44
              *U.S.  GOVERNMENT  PRINTING  OFFICE:  1991—517-003/47034

-------