FINAL
             Umted States                               ECAO-CIN-426
             Environmental Protection                          Fahnuru
             Agency                                 reuruary.
IvEPA      Research and
             Development
             DRINKING WATER CRITERIA DOCUMENT
             FOR TOXAPHENE
             Prepared for

             OFFICE OF DRINKING WATER
             Prepared by

             Environmental Criteria and Assessment Office
             Office of Health and Environmental Assessment
             U.S. Environmental  Protection Agency
             Cincinnati, OH  45268

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                                  DISCLAIMER

    This  document  has  been  reviewed  In  accordance  with  the  U.S.  Environ-
mental  Protection  Agency's   peer   and administrative   review  policies  and
approved  for  publication.   Mention of  trade  names  or commercial  products
does not constitute endorsement or recommendation for use.
                                       11

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                                   FOREWORD  •


    Section  1412  (b)(3)(A)  of  the  Safe  Drinking Water  Act,  as  amended  In
1986,  requires  the Administrator  of the Environmental Protection  Agency  to
publish  maximum  contaminant level  goals  (MCLGs)  and  promulgate  National
Primary  Drinking  Water   Regulations  for  each  contaminant,   which,   in  the
judgment of  the Administrator,  may  have  an adverse  effect on public health
and  which  1s known  or anticipated  to  occur  1n  public  water  systems.   The
HCLG  1s  nonenforceable and  1s  set  at  a  level  at which no known  or  antici-
pated  adverse  health effects   In   humans  occur  and which   allows   for  an
adequate margin  of  safety.  Factors considered  In setting the  MCLG  Include
health effects data and sources  of exposure other than drinking water.

    This document provides  the  health  effects  basis  to  be  considered  In
establishing  the  MCLG.  To  achieve  this  objective,  data  on pharmacoklnetlcs,
human  exposure,  acute and  chronic  toxldty  to animals and humans,  epidemi-
ology and mechanisms  of toxldty  are evaluated.   Specific  emphasis Is placed
on  literature  data  providing  dose-response   Information.  Thus,  while  the
literature search  and evaluation performed  1n support of this  document  has
been comprehensive, only  the reports considered most  pertinent  In  the deri-
vation of  the MCLG are cited 1n  the document.  The  comprehensive  literature
data base  1n support   of  this document  Includes  Information published  up  to
1984;  however,  more   recent  data  may  have   been  added   during the  review
process.

    When adequate health  effects  data exist.  Health  Advisory  values  for less
than   lifetime  exposures  (1-day,   10-day  and   longer-term,   -10X   of   an
Individual's  lifetime) are  Included  1n  this document.  These  values  are  not
used 1n  setting the MCLG,  but  serve as  Informal  guidance to  municipalities
and  other  organizations   when  emergency  spills  or  contamination  situations
occur.
                                                 Michael B. Cook
                                                 Director
                                                 Office of Drinking Water
                                      111

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                             DOCUNENT DEVELOPMENT
Annette M. Gatchett, B.S., Document Manager
Environmental Criteria and Assessment Office, Cincinnati
U.S. Environmental Protection Agency

Helen H. Ball, M.S., Project Officer
Environmental Criteria and Assessment Office, Cincinnati
U.S. Environmental Protection Agency
Authors

Shane Que Hee, Ph.D.
Martha Radlcke, Ph.D.
University of Cincinnati
Department of Environmental Health
Cincinnati, OH
Reviewers
                     M.S.
Randall J.F. Bruins,
Frank M1nk, Ph.D.
Environmental Criteria and
  Assessment Office, Cincinnati
U.S. Environmental Protection Agency

William Marcus, Ph.D.
Jennifer Orme, M.S.
Office of Drinking Water
Washington, DC

Fumlo Matsumura, Ph.D.
Pesticide Research Center
Michigan State University
East Lansing, MI
(Contract 68-03-3234; Eastern
  Research Group)
William E. Pepelko, Ph.D.
Carcinogen Assessment Group, OHEA
U.S. Environmental Protection Agency
Washington, DC

James WUhey, Ph.D.
Sir Frederick Banting Research Center
Bureau of Chemical Safety
Ottawa, Ontario
Canada
(Contract 68-03-3234; Eastern
  Research Group)

James 0. Pierce
University of Southern California
Cooperative Agreement
No. CR 813569-01-0

Editorial Reviewers

Erma Durden, B.S.
Judith Olsen, B.A.
Environmental Criteria and
  Assessment Office, Cincinnati
U.S. Environmental Protection Agency
Document Preparation
Technical Support Services  Staff:   C.  Cooper, P.  Daunt,  C.  Fessler
B. Zwayer, Environmental Criteria and Assessment Office, Cincinnati
                                                                      K. Mann,
                                      1v

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                              TABLE  OF  CONTENTS
  I.   SUMMARY ...........................      i.i

 II.   PHYSICAL AND CHEMICAL PROPERTIES ...............     II-l

      PRODUCTION AND USAGE .....................     II-8
      SPECTROSCOPIC PROPERTIES ...................     II-9
      ANALYSIS ......  .....................     11-12
      SUMMARY ...........................     II-15

III.   TOXICOKINETICS ........................    III-l

      ABSORPTION ............... ' ...........    III-l
      ORAL .............................    III-l
      INHALATION ..........................    III-5
      DERMAL ............................    III-5
      DISTRIBUTION .........................    TII-5
      DISTRIBUTION IN ANIMAL TISSUES ................    III-6

           Birds ......................  .....    III-6
           Mammals .........................    III-9

      DISTRIBUTION IN HUMAN TISSUES .  ...............    111-19
      METABOLISM ..........................    111-19
      ELIMINATION .........................    111-25
      SUMMARY ...........................    111-31

 IV.   HUMAN EXPOSURE ........................     IV-1

  V.   HEALTH EFFECTS IN ANIMALS ..................      V-l

      ACUTE AND SUBACUTE TOXICITY ...........  .  .....      V-l
      CHRONIC TOXICITY .......................      V-18
      ENZYME EFFECTS ........................      V-26
      TERATOGENICITY AND REPRODUCTIVE  EFFECTS ...........      V-37

           Mammals .........................      V-37
           Birds ..........................      V-44

      MUTAGENICITY .........................      V -47
      CARCINOGENICITY .......................      V-50
      SUMMARY ..... .....................  .      V-65

 VI.   HEALTH EFFECTS IN HUMANS ...................     VI-1

      ACUTE TOXICITY ........................     VI-1
      SUBCHRONIC AND CHRONIC TOXICITY  ...............     VI-5
      SUMMARY ......................  .....     VI-7

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                                                                       Page

 VII.  MECHANISMS OF TOXICITY	    VII-1

       INTRODUCTION	    VII-1
       ENZYMATIC EFFECTS 	    VII-2
       MUTAGENIC ACTIVITY	    VII-7
       SUMMARY	    VII-8

VIII.  QUANTIFICATION OF TOXICOLOGICAL EFFECTS 	   VIII-1

       INTRODUCTION. ...  	   VIII-1
       NONCARCINOGENIC EFFECTS 	  .  	   VIII-6
       QUANTIFICATION OF NONCARCINOGENIC EFFECTS 	  ,   VIII-12

            Derivation of 1-Day HA 	   VIII-12
            Derivation of 10-Day HA	VIII-13
            Derivation of Longer-Term HA 	   VIII-14
            Assessment of Lifetime Exposure and Derivation
            of a DWEL	VIII-14

       CARCINOGENIC EFFECTS	VIII-14
       QUANTIFICATION OF CARCINOGENIC EFFECTS	VIII-20
       EXISTING GUIDELINES, RECOMMENDATIONS AND STANDARDS	VIII-23
       SPECIAL GROUPS AT RISK	VIII-26

            Sensitive Subpopulatlon	VIII-26
            Interactions 	   VIII-27

  IX.  REFERENCES.	     IX-1

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                               LIST OF TABLES

No.                                  Title                            Page

 II-l   Nomenclature, Indexing Terms and Synonyms Currently
        Used for Toxaphene	     II-3

 II-2   Identified Toxic Components of Toxaphene	     II-4

 II-3   Physical Properties of the Degradation Products
        of Toxaphene. .	     11-6

 II-4   Chemical Information Regarding Toxaphene Mixtures 	     11-10

 II-5   Examples of Analytical Methods for Toxaphene	     11-13

III-l   Absorption Data  for All Routes for Toxaphene	    III-2

III-2   Distribution of  Toxaphene Continuously Administered
        Orally to Black  Ducks from Birth by Mixing Toxaphene 1n
        Propylene Glycol,  which Constituted IX of the Duck Mash .  .    111-10

III-3   Percent Uptake of  Radioactivity 1n Various Rat Tissues
        and Organs Following a Single Dose of "Cl-Toxaphene
        (20 mg/kg) Administered 1n 0.5 ml Peanut 011/Green
        Acacia	    111-12

III-4   Distribution of  Radioactivity 1n Various Rat Tissues
        and Organs at Day  14 Following a Single Dose of
        36Cl-Toxaphene,  14C-Toxaphene, Toxicant A and B
        Administered 1n  Corn 011 by Stomach Intubation	    III-14

III-5   Distribution and Metabolism of Toxicant B (51775-36-1)
        at 7 and 72 Hours  After Stomach Intubation of Sprague-
        Dawley Rats at 3.1 mg/kg bw	    111-16

III-6   Distribution of  Radioactivity 1n Various Female Sprague-
        Dawley Rat Tissues and Organs Following a Single Dose of
        2.6 ing 14C-Toxaphene/kg bw Dose to Four Pregnant Rats
        Administered In  Olive 011 (0.1 mi) by Oral Gavage 	    111-18

III-7   Identified Metabolites of Toxaphene Components	    111-22

III-8   Elimination of 14C-Toxaphene, Toxicant A and
        Toxicant B Administered to Sprague-Dawley Rats
        by Stomach Intubation 1n Corn 011 Carrier 	    111-28

III-9   Experimental or  Calculated Half-Life of Toxaphene
        Elimination 1n Various Species	    111-32

  V-l   Acute Oral Toxlclty of Technical Toxaphene
        (CAS RN 8001-35-2) to Laboratory Mammals.	      V-2

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No.              '                    Title                            Page
  V-2   Acute Dermal LDsg Values for Toxaphene 1n Laboratory
        Mammals	      V-5
  V-3   Acute Toxlclty of Technical  Toxaphene Components
        Administered IntraperHoneally to Mice	      V-6
  V-4   Residue Concentrations In Dead Birds  and Animals	      V-10
  V-5   Acute and Subchronlc Oral Toxldty of Toxaphene	      V-13
  V-6   Chronic Toxlclty of Toxaphene to Laboratory Mammals
        at Low Dietary Levels	      V-19
  V-7   National Cancer Institute Chronic Feeding Study 	      V-22
  V-8   Incidence of Tumors In Male  Rats Fed  Toxaphene	      V-53
  V-9   Incidence of Tumors In Female Rats Fed Toxaphene	      V-54
  V-10  Incidence of Tumors 1n Male  Mice Fed  Toxaphene  In
        the Diet	      V-56
  V-ll  Incidence of Tumors In Female Mice Fed Toxaphene
        1n the Diet	      V-57
  V-12  Survival of Mice Fed Toxaphene 1n the Diet	      V-59
  V-13  Hepatocellular Tumor Incidence In B6C3F1 Mice Fed
        Toxaphene In the Diet	      V-61
  V-14  Variation 1n Doses of Toxaphene Used  1n NCI (1979)
        Study of CarclnogenlcHy	      V-69
 VI-1   Case Studies of Toxaphene Poisoning 1n Humans 1n which
        Ingestlon Was the Primary Route of Entry	     VI-3

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                             LIST OF  ABBREVIATIONS
ATP
bw
CNS
DNA
dw
DWEL
GC
GI
HA
l.p.
1.v.
LD50
LOAEL
MEL
MFO
NADPH
NOAEL.
NOEL
ppm
RfD
s.c.
TLC
TLV
Toxicant A
Toxicant A-l
Toxicant A-2
Toxicant Ac
Toxicant B
TWA
ww
Adenoslne trlphosphate
Body weight
Central nervous system
DeoxyMbonuclelc add
Dry weight
Drinking water equivalent level
Gas chromatography
Gastrointestinal
Health advisory
IntrapeMtoneal
Intravenous
Lethal dose to 50% of the recipients
Lowest-observed-adverse-effect level
Minimal effect level
Mixed function oxldase
N1cot1nam1de adenlne dlnucleotlde phosphate dehydrogenase
No-observed-adverse-effect level
No-observed-effect level
Parts per million
Reference dose
Subcutaneous
Thin-layer chromatography
Threshold limit value
Subfractlon of toxaphene separated chromatographlcally
A toxic component of Toxicant A
A toxic component of Toxicant A
A contaminant of Toxicant A
Subfractlon of toxaphene separated chromatographlcally
Time-weighted average
Wet weight
                                      1x

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                                  I.   SUMMARY

    Toxaphene  1s  a  broad  spectrum,  chlorinated  terpenold  pesticide  Intro-
duced  In  the  United States as a  contact  Insecticide  In 1948.   Together  with
methoxychlor,  toxaphene  has  been the  chief  replacement for DOT  when  use of
DOT was banned.  On  May  25,  1977, the U.S.  EPA Issued a notice of rebuttable
presumption  against registration  and continued  registration  of  pestlcldal
products  containing  toxaphene.   A further notice  was  Issued  1n  the Federal
Register  In  1982.   Registration  for  use after  December  31,  1983 for scabies
treatment  of  beef  cattle  and sheep  1n  vat  dips and  spray dips  was  still
allowed  under  specified  conditions.   Toxaphene  use   to  control  sporadic
Infestations  of  army worms,  cutworms and grasshoppers  on cotton,  corn  and
small grains was also permitted  under specific  conditions.   Uses for control
of  mealybugs  and  pineapple  gummosIs  moths   on pineapples,  and weevils  In
bananas are  allowed only  In the Virgin  Islands  and  Puerto R1co.   Sale  and
distribution of existing toxaphene  stockpiles  was  allowed  until December  31,
1986 with  the use of toxaphene  under limited  conditions  only.   In addition
to  the  previous  uses,  It may be  applied  to  soybeans  and peanuts for sickle-
pod control.   Its sale and  distribution  for  use as an  Insecticide 1n no-till
corn and dry and southern  peas was  permitted  until December 31, 1986.   These
administrative decisions were made  as a result of the  toxic effect of  toxa-
phene on wildlife,  aquatic  organisms,  nontarget organisms,  and the potential
oncogenldty of toxaphene In humans.

    A  variety of  guidelines or  standards  have  been  published  to protect
against toxaphene  1n aquatic media.   The national Interim primary drinking
01990                                1-1                             02/25/87

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water  standard  Is  5  yg/i.   The  FOA mandated  level   for  bottled water  Is
also 5 yg/l.  These standards are based upon organoleptlc effects.

    The  water  quality criterion  for  the  protection  of  human  health  was
originally  set  at  0.5  wg/l  for  a  10"5  cancer  risk   level.   This  cri-
terion was  subsequently  revised  to  7.1 ng/l  for  a risk of 10"5.   If  con-
sumption  of aquatic  organisms  1s considered  alone, the  corresponding  risk
level  1s  7.3 ng/l.   These criteria  were based  on the  Induction  of  hepato-
cellular tumors In mice.

    Toxaphene  1s  a mixture  {average  molecular weight  414) of at  least  177
compounds,  mostly chlorinated  camphenes,  which  has  been used  as  a  broad
spectrum  contact  pesticide  (e.g.,  In  soybean and  cotton  fields,  and  for
ectoparasites).   It Is  produced  Industrially by the  reaction  of camphene  and
chlorine  1n  carbon  tetrachlorlde  solvent   1n  the  presence of  ultraviolet
light  to  produce  a  product  of  highly variable  quality.  Toxaphene  can  be
provided as  a 40X dust concentrate,  as a liquid  formulation  containing  10%
xylene, or  as  emulslflable  concentrates.    It  1s extensively  co-formulated
with other pesticides  (e.g., DDT, parathlon,  endrln).

    The vapor  pressure,  densUy,  solubility  and  other  physical  properties
vary according  to the quality of the product.  Toxaphene Is  nonpolar  and Is
soluble  In  nonpolar  solvents;   It  1s  less  soluble In polar  solvents.   It
adsorbs to  soils, and partitions readily Into  n-octanol.  Because toxaphene
1s aliphatic,  It  Is a  poor  absorber  of ultraviolet light above wavelengths
of 250  nm.   Gas  chromatography  with  electron  capture  detection  or  chemical
01990                                1-2                             02/25/87

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 1on1zat1on  mass  spectrometrlc detection  are  commonly used to  measure  toxa-
 phene  levels  In  biological  samples.   In a  few  cases,  'H- and  13C-nuclear
magnetic resonance techniques have  been  used  to assess  the "fingerprints"  of
 toxaphene fractions.

    Toxaphene  readily  dehydrochloMnates  on  heating above  120°C,  but  the
toxaphene subfractlons  Toxicants  A  and B appear  to  be  stable  on  gas  chroma-
tography.   DehydrochloMnatlon  also occurs  In  the presence of alkali  (used
as an  analytical  method),  of reduced hematln, or  Iron  compounds.   Reductive
dechlorlnatlon  occurs  on   ultraviolet   light  Irradiation  (230-290  nm)  of
toxaphene In organic  solvents, preferentially at the C. and Cr  positions.

    Most of  the analytical  techniques  used  to detect  toxaphene  metabolites
have  employed  thin-layer   chromatography  as  the   end  step.  Toxaphene  1n
environmental samples  and  tissues  often does  not have  the  chromatographk
pattern  of  a  toxaphene  standard  so that  qualitative  Identification by gas
chromatography  occurs  primarily   through  retention time;  quantltatlon  1s
generally  accomplished  by   the  method  of  alkaline  dehydrochlorlnatlon,
extraction and chromatography (Gomes method,  detection  limit  1  ng).   Metabo-
lites  are generally  Isolated by  thin-layer  chromatography  or  liquid  chroma-
tography.

    Though no quantitative  data  are available, the  symptoms of toxicosis  as
well as  the  presence of residues and metabolites  after exposure  1n animals,
birds  and humans  Indicate that toxaphene can be  absorbed  through  the  skin,
the lungs, and the gut.
01990                                1-3                             02/25/87

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    In  White  Leghorn chickens,  accumulation of toxaphene residues  occurs  at
doses  of  >12.5 mg  toxaphene/kg  bw/day.   In ducks,  brain  residues  were
detected at concentrations >10  ppm  1n  the feed.   Extensive  dechloMnatlon  of
toxaphene  by  chlorinated organic residues  occurs  mostly  1n the  fat.   Doses
less  than  those resulting 1n accumulation  can be tolerated for  months  with
IHtle effect.

    In mammals,  storage  of  toxaphene 1n  the fat  has  been reported  In  sheep,
steers  and  dairy  cows.   Accumulation  of  toxaphene  1n  sheep  and  steers
occurred at  administered concentrations  between  10  and 100 mg  toxaphene/kg
feed/day.   In  contrast,  dogs appear to  store  toxaphene  derivatives  In  the
brain.   In male or  female  rats, fat  levels  of  toxaphene  were  relative  to
concentrations  >21  ppm  In  the  diet.   Studies  using   labeled  toxaphene
revealed  rapid  dechlorlnatlon   and  subsequent  elimination  of  toxaphene.
Toxicant A  and Toxicant  B  at doses <19  mg toxaphene/kg  bw administered  by
gavage to male  rats  using corn  oil  as  a carrier.   Toxaphene derivatives  were
measured  1n  the  blood,  fat,   liver   and   kidneys.   When  male  rats  were
chronically dosed  at 2.4 mg  toxaphene/kg bw/day, plateau levels were  found
In the  liver  and brain after 1, 3  and  6  months.   Transplacental  transfer  as
well  as  blood-brain  barrier  transfer occurred In rats.  The  adrenal  gland,
carcass, cecum  and abdominal  fat of female  rats  contained  toxaphene deriva-
tives 3  days  after  an acute  dosing with 2.6  mg  toxaphene/kg  bw.  Extensive
dechlorlnatlon  and rapid elimination of  metabolites  also occurred  in  guinea
pigs, hamsters, rabbits,  mice and monkeys.
01990                                1-4                             02/18/87

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    In  rats,  successful  detoxification  appeared to require NAOPH and oxygen.
Toxicants  A  and B undergo reductive dechlorlnatlon, dehydrochlorlnatlon, and
vicinal  chloride elimination.   Involvement  of  the MFC.  system  was  suggested
by  the observation  of Type  I   binding  spectra with  the  hepatic  cytochrome
P-450  of  rats,  mice,  sheep  and  rabbits, as well as the demonstrated enhanced
toxldty  to  mice of toxaphene 1n  the presence  of  plperonyl butoxlde.  There
also  appeared  to  be  a  detoxification  pathway dependent  upon  glutathlone.
Toxicant  8 resisted  metabolism  more than Toxicant C.   Glucuronlde  (major)
and sulfate  (minor) conjugates   have been  detected as  metabolites.   Toxicant
C also produced one primary  and four   secondary alcohols,  whereas  reductive
dechlorlnatlon  predominated  for  Toxicant  B.   These  data  suggested  that
toxaphene  components  were  not  necessarily metabolized  at  the  same  loci  In
the mlcrosomal  system.

    Elimination  of  products derived from  toxaphene  has  been  demonstrated to
occur  In  the  feces, urine and expired  air of  rats.   The fecal  route appears
to be  slightly more predominant than  the urinary route.   Milk  and  eggs may
also  contain  residues.   The  species order  of decreasing  elimination  effi-
ciency  for Toxicant B Is as  follows:   monkey,  rat,  hamster > mouse,  rabbit,
guinea pig > chicken.

    Acute  oral  exposure  to  toxaphene  gave  LD.-  values ranging  from 7.5-10
mg/kg  bw  1n  female monkeys  to  75-500  mg/kg   1n  rabbits.   The  LD5Q values
resulting  from  dermal  application  of  toxaphene  are  generally  higher  than
those  observed  after  oral  exposure; they range from  -250  mg/kg bw  to -4000
mg/kg  bw  with  both  values  being  reported for rabbits.   The vehicle used with
the  pesticide  may  affect   the  toxic  response.   Specific  components  of

01990                                1-5                             02/25/87

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 technical  grade  toxaphene are  more toxic  than  the complex mixture  and,  as
 reported  for  mice,   L05Q values   for  several  components   varied  from  2.5
 to >100  mg/kg bw.   Brain levels   of  toxaphene  may  be  Indicative of  acute
 tox1c1ty.   In  swine,  >4 mg/kg ww  (brain  tissue)  constituted a lethal level,
 and 2 mg/kg was associated with clinical signs of toxldty.

    Subchronlc oral doses  of  toxaphene  In  laboratory animals resulted In few
 clinical  signs  of poisoning.   Lethality  responses  varied  In  mice fed  1280
 ppm toxaphene  In  the diet for  6 weeks.  No  observable  adverse  effects  were
 seen  In  rats  following  1ngest1on  of  feed containing  189 ppm  toxaphene..   At
 concentrations <189  ppm In the  diet,  subcllnlcal  lesions  such  as  decreased
 bile  production  and  questionable  liver  pathology  were reported.    In  mice
 receiving 100  or  200  ppm toxaphene 1n  the  diet,  humoral antibody production
 (IgG  antibody  formation)  was  suppressed;  however,  cell-mediated   Immune
 responses were not affected.

    Inhaled  toxaphene mist  concentrations as  high  as  500 mg  toxaphene/m3
 for 3 weeks  caused  no mortality 1n rats and  rabbits;  but at 12  mg toxaphene
dust/m3 for 3 months, mortality occurred 1n rats, dogs and guinea pigs.

    Long-term exposures  of animals to  dietary levels of toxaphene are  sum-
marized  In  Chapter  V.    In  a  number   of  studies,  no adverse  effects  were
reported  among the  parameters  monitored  In each  experiment.   In  several
 lifetime  studies  In rats, no effects  were  reported  for dietary concentra-
tions  between  10  and 100  ppm toxaphene;  other studies  at  100 ppm toxaphene
 level   reported  liver pathology.    The  lowest dietary level  producing  liver
damage  In  rats was  5 ppm for  2  years.   In  addition to liver  pathology,  a


01990                                1-6                             02/25/87

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variety  of  adverse  effects  were  reported  from  both  Vn  vivo  and in  vUro
studies  and  Included  kidney  pathology,  decreased  bile  flow,  decreased
survival,  Inhibition  of  Intestinal   transport   of  glucose,  Inhibition  of
gluconeogenesls.   Inhibition  of   mltochondrlal   enzymes,   Inhibition   of
Nah/Kf-ATPase   brain  and   kidney  activity,   and   Inhibited   Mg2f-ATPase
activity In mouse kidney, liver and brain.

    Toxaphene  Induced  the  mlcrosomal  MFO  system;  treatment  of  animals  j_n
vivo with a cytochrome P-450 system Inhibitor Increased toxaphene toxldty.

    No  effects   on  parental  animals   or  offspring  were  noted  1n a  throe-
generation study  In  which  Sprague-Oawley rats were  fed  dietary  levels  of  25
or 100  ppm  toxaphene.   In  a study  using  CO  rats,  maternal  mortality  was 31%
at  35  mg  toxaphene/kg  bw.   There  was  also  a dose-related  decrease  1n
maternal weight  gain and fetal weight  (15,  25 or 35  mg/kg  bw).   No  adverse
effects were reported  In the offspring of mixed-strain white rats dosed with
4 mg/kg bw during  the entire pregnancy.  In  hamsters given the  same  dose,
toxaphene was reported to be teratogenlc but  the anomaly was not Identified.

    More sensitive endpolnts In  rats were examined by several  Investigators.
Toxaphene, at  12 mg/kg  bw  administered  orally  to  pregnant rats, depressed
chollnesterase  activity  In  fetal cardiac   neural   structures  and  delayed
cardiac  neural   differentiation.    At   25 mg/kg  bw,   the  only   significant
differences noted  were decreases  1n alkaline  phosphatase  activity and  total
protein  1n  the  kidney.   There was no effect  at 12.5 mg/kg  bw  on  the rat
01990                                1-7                             02/25/87

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kidney.   In  two  separate  experiments  there  was  IHtle evidence of behavorlal
abnormalities  1n  the  offspring of toxaphene treated  dams  except  for  delayed
righting  ability  (6 mg/kg and 50 vg/kg bw).

    Toxaphene  given  to pregnant mice  at  15 and  25  mg/kg bw  had  no  adverse
effect  on  parameters  examined  In  offspring.    However,  a  dose  of  35  mg
toxaphene/kg bw  Induced encephaloceles In  five  Utters of 90  treated  dams.
In  a  5-  to  6-generat1on  study  where  toxaphene  (25  ppm) was  given  1n  the
diet,  little  or  no adverse  effects  were  observed In mice.   No  effects  were
noted  on  the  anatomical  development  of  the  fetal  guinea pig  from pregnant
females given  15 mg/kg bw from days 21-35  of gestation.

    Toxaphene  appears  to  have a  low  teratogenlc potential unless  doses  are
large enough to  Induce  maternal  toxIcHy.   In  the studies reported here,  the
lowest  dose  producing an  effect was  50  vg/kg bw  (rats)  given  In the  diet
from day  5  of  gestation through termination of  the  study.  Effects Included
significant  decreases  1n  overall  swimming  ability  and  righting  reflex  of
young pups when  toxaphene was  administered  before postnatal  day 16.  In  this
study,  however,  the  precise  dose  to  the  offspring  Is uncertain  since  they
will receive toxaphene transplacentally, 1n the milk and In the feed.

    Toxaphene  1s  mutagenlc   1n  the  Salmonella/mlcrosomal reverse mutation
assay using  strains  TA98 and  TA100.   Results  of assays  of  toxaphene  compo-
nents  for their  mutagenlc potential  Indicated that  mutagenlclty  resided  in
the polar fraction.   Mutagenlclty was  decreased  by  the  addition  of  a  liver
mlcrosome, S9, an  active  HFO (these results support  the  Jm  vivo observation
that  Inhibitors   of   cytochrome  P-450  Increased  toxldty).   The  mutagenlc
activity  of commercial preparations varies.
01990                                1-8                             02/18/87

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    Studies  of  toxaphene cardnogenkHy are  reported  In  Chapter V.   In  an
NCI carcinogenic study many  doses were  lowered because  of  the  overt  toxlclty
seen  1n  rats and mice  Ingesting  feed containing toxaphene.   The concurrent
numbers  of  control  animals  were  low (10/sex/group) and historical  controls
were used 1n order  to match  the  number  of treated animals  per  group  (50/sex/
group).   The range  of doses  given  to the animals  made H difficult  to  use
time-weighted  averages  of doses  as  a  basis  on which  to  estimate  risk  for
humans.   It  was  concluded that under  the conditions of the bloassay,  toxa-
phene  was  carcinogenic   1n male  and  female B6C3F.  mice,  as evidenced  by  an
Increased Incidence of hepatocellular carcinomas 1n  a dose-related manner.

    In a  separate  study using B6C3F,  mice of  both  sexes,  fed 7, 20  and  50
mg  toxaphene/kg  diet,  the carclnogenlcUy  of toxaphene was demonstrated  at
much lower doses.   After 18  months  of  toxaphene  Ingestlon  and 6  months on a
control  diet,   significantly  Increased  Incidences  of  hepatocellular  car-
   t,
dnomas  were  detected   1n males.   The  response 1n females  was less  pro-
nounced.  Another study  reported only  one tumor,  a  subcutaneous neurosarcoma
In rats exposed to 1000 ppm toxaphene 1n the diet.

    Based on the positive carcinogenic  evidence of  toxaphene from  the  NCI
mouse  and  rat  studies and the  other mouse study,  1t  can  be  concluded that
toxaphene  1s carcinogenic 1n at least  two  species  of laboratory  animals.
Using  the EPA  criteria  for classifying cardnogenldty  data,   the  animal
evidence would be considered   "sufficient".
01990                                1-9                             04/02/87

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    Toxaphene  has  caused  convulsions and nausea  In  humans  when exposure has
occurred  by  Ingestlon,  skin  contact  or  Inhalation.   The  acute  oral  L05Q
was  estimated  from accidental  poisonings  to  vary  between 29  and  100  mg
toxaphene/kg  bw.   Mixtures of  toxaphene  with  DOT (2:1,  by  weight) are more
acutely  toxic  than either  component alone.  The  estimated  hazardous dermal
dose  (applied  to  the  skin) 1s  -660  mg/kg bw for  liquid toxaphene.  Allergic
bronchopneumonla  has   been  observed  after acute  Inhalation  of  toxaphene
sprays.  Women  have been  shown to suffer  a significant Incidence of chromo-
somal aberrations In lymphocytes.

    Dermal  doses  of 300 mg/day  for  30 days, and  Inhaled doses  of 0.4 mg/m3
for  10  minutes on each  of   15  days  and  250  mg/m3  for   30  minutes  (-41
mg/day) on each of  13 days caused no atypical subjective or  clinical effects.

    Studies  In  the  workplace  are  confounded  because  exposure  to  many
chemicals had occurred  1n  all  the  reported epidemlologic  studies.  There are
no  porphyrlnogenlc  or  sympathotonlc   effects  mentioned  In   the  available
literature.

    Human  data  are  Inadequate to  assess  the  carcinogenic   potential  for
toxaphene.

    Animal  studies  were used  to derive  the 1- and 10-day HAs  as  a result of
the  lack of  human data.   The  recommended 1-day  health advisory  (HA)  for
toxaphene  In  drinking  water  1s 0.5 mg/l for  a  10  kg child.   A  NOAEL  of  5
mg/kg bw based  upon the onset  of convulsions In dogs receiving a  single dose
of  toxaphene  was  used to  derive  this  HA.  The  recommended  10-day  HA for
01990                                1-10                            02/18/87

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toxaphene  In  drinking water  Is  0.04 mg/i for  a  10 kg child.  A  LOAEL  of 4
mg/kg  bw  based upon  minimal  kidney  and liver pathology  1n  dogs  was  used to
derive this MA.

    There  are no  acceptable  studies   1n  the available  literature  for  the
derivation of a longer-term HA or a  lifetime DWEL.

    The recommended  criterion for chronic exposure  to  toxaphene  1n drinking
water, not  considering  Intake from  other  sources,  Is  3.1, 0.31  and  0.031
vg/4,   to   maintain   Individual   cancer   risk  less  than   10~4,   10~5   and
10~6,  respectively.   This  criterion Is  based  upon  an   Increase  1n  hepato-
cellular   tumors  1n male  mice fed 7,  20 and 50  ppm toxaphene In  the  diet.
The  sufficient  level  of  carcinogenic  evidence  In  experimental  animals,
together  with  the  Inadequate  level of  human  evidence meets the IARC Group 28
criteria   for  weight  of  evidence  that  a  chemical   Is  likely  to  be  a  human
carcinogen.  The assignment of a  Group  28 designation by IARC means that the
chemical    1s   probably  carcinogenic   1n  humans.    Applying  the  criteria
described  In  the  U.S.  EPA's  guidelines  for  assessment  of  carcinogen  risk,
toxaphene may  be classified as Group B2:   probable  human carcinogen,  meaning
there  1s   Inadequate evidence  from human studies and sufficient evidence from
animal studies.
01990                                1-11                            05/12/87

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                     II.   PHYSICAL AND CHEMICAL  PROPERTIES

    Toxaphene  Is  a  broad spectrum,  chlorinated  terpenold  pesticide.  Intro-
duced  as  a  contact  Insecticide  Into  the  United  States  In 1948  (Brooks,
1974).   Together  with methoxychlor,  1t was  and has been  the chief  replace-
ment  for  DOT  after  DOT was banned.   On May  25, 1977,  the U.S.  EPA  Issued  a
notice  of rebuttable presumption  against  registration and  continued  regis-
tration   of  pestlddal  products  containing  toxaphene  (Federal   Register,
1977).   A further  notice  was  Issued  In the  Federal  Register (1982).   Regis-
tration  for use after December  31,  1983  for  scabies  treatment of beef  cattle
and  sheep In  vat  dips  and spray dips was  still  permitted  under  specified
conditions.  Toxaphene  use to  control  sporadic Infestations of army  worms,
cutworms  and  grasshoppers  on  cotton,   corn  and  small   grains   was  also
permitted  under  certain  restrictions  (Federal  Register,  1982).   The  use  of
toxaphene to control mealybugs,  gummosls moths  on  pineapples, and  weevils  In
bananas  1s allowed only 1n  the  Virgin Islands and  Puerto  R1co.   Depletion  of
existing  toxaphene   stockpiles  was  allowed   until  December  31,  1986  under
limited  conditions   only.   In  addition  to   the  previous  uses,   It  may  be
applied  to  soybeans  and  peanuts for slcklepod  control.   Its sale and  dis-
tribution  for  use as  an  Insecticide  In  no-till corn  and dry  and  southern
peas was  permitted until  December 31, 1986  (Federal Register, 1982).   These
administrative  decisions   were   made  as  a  result  of  the  toxic  effect  of
toxaphene on wildlife, aquatic  organisms,  nontarget  organisms, and potential
oncogenlclty of toxaphene 1n humans.

    Toxaphene can be  produced  Industrially by the  reaction between technical
grade  camphene  and  chlorine   (In  carbon  tetrachloMde  solvent)   In  the


02000                                II-l                             02/25/87

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presence of  ultraviolet  light  (mercury arc radiation) for  15-30  hours  until
the  final  product  has  a chlorine content  between  67  and  69% (SHtlg,  1977).
The chemical quality of the final product 1s highly variable (IUPAC,  1979).

    The many synonyms of toxaphene are  provided  In Table  II-l.   Toxaphene Is
known  as   chlorophene  or   polychlorocamphene   In  the   Russian   literature
(Melnlkov,  1971).  Synonyms, chemical names and  Indexing  terms  for  the  major
toxic  components  of  toxaphene  are  given  1n Table  II-2.   The  most  toxic
Ingredients  to mammals  1n  technical  toxaphene are Toxicant  A  and Toxicant  B
(Table  II-2).  each  consisting  of  -2-6X  of the mixture  (Metcalf,  1981;
Khalifa et al.,  1974).  Toxicant A consists of  three  toxic components,  Toxi-
cant A-l  (CAS  No.  58002-18-9) and  Toxicant A-2 (CAS No.  58002-19-0),  and  a
contaminant, Toxicant  Ac  (CAS  No.  66860-80-8)   (Chandurkar  et  al.,  1978).
Synonyms,  chemical  names  and  Indexing  terms  for  the major metabolites  and
degradation products of toxaphene are provided 1n Chapter  III.

    Since  the  technical grade  product  1s produced by  free radical  reactions
Initiated  by ultraviolet  light, toxaphene  Is   a  complex  mixture  of  poly-
chlorinated  camphenes  and  bornanes with  an  average  empirical  formula  of
C,nHinClQ,   and  an average  molecular   weight of  414.   In  fact,  over  177
 i u  i u  o
Incompletely characterized components  have been  separated (Holmstead  et al.,
1974).   Technical   grade   toxaphene  1s  an amber,  waxy  solid with  a  mild
terpene odor, a  softening  range  of 70-95°C, a vapor  pressure of 0.17-0.40 mm
Hg at  25°C,  and  a  density  of  1.66  g/ml  at 27°C  (Brooks,  1974).   The  solu-
bility  1n  water  has been  variously  reported as  -3 mg/i  (Brooks,  1974)  and
0.5  mg/i  at 25°C  (Paris  et al..  1977).   Differing  values for  vapor  pres-
sure,  solubilities and other  physical  constants  are to be  expected  since


02000                                II-2                            02/25/87

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                                  TABLE II-l
    Nomenclature,  Indexing  Terms  and  Synonyms Currently Used for Toxaphenea
       CAS RN 8001-35-2°
           Empirical  Formula:
Toxaphene (DOT)C (SCI & 9CI)d«e
Agrlclde maggot killer (f)
Alltex
ATHox
Camphechlor
Camphochlor
Chem-Phene
Chlorinated camphene
Chlorinated camphene, tech
Chlor chem t-590
Compound 3956
Estonox
ENT 9,735
Fasco-Terpene
Genlphene

Gy-Phene
Hercules 3956
            Kamfochlor
            M  5055
            Mellpox
            Motox
            NCI-C 00259
            Octachlorocamphene
            Penphene
            Phenaclde
            Phentox
            Polychlorcamphene
            Polychlorocamphene
            PCHK
            Strobane T
            Synthetic 3956
            Technical chlorinated
            camphene  (67-69%) chlorine)
            Toxadust
            Toxafeen  (Dutch)
            Toxakll
            Toxaphen  (German)
 Source: SANSS data base, June 1983
bChem1cal Abstracts Service Registry Number
C0epartment of Transportation
dE1ghth Collective Index, Chemical Abstracts
eN1nth Collection Index, Chemical Abstracts
02000
II-3
02/03/87

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o
ro
O
                                                                            lABlt 11-2

                                                             Identified loxtc Components of loxaphene
          CAS RN
             tv
                                      Cheated! Abstracts
                               (9CI)
         Synonyas
    Reference
         51775-36-1
         C10HnCl7
         52819-39-3
         57028-55-4
         57981:- 30-3
         58002-18-9
         CIOHIOC'8


         58002-19-0
         66860-80-8
         70940-13-5
Btcyclo|2.2.1)heptane.2.2.5.6 tetrachloro
1.7.7.-trls(chloro«elhyl)-.<5 endo.6 exo)
Blcyclo(2.2.1]heptane.2.3.3.5.6-pentachloro-
7.7-bls(chloroM>thyl)-l (dtchloro«ethyl)-
(2-endo.5-exo.6-exo)-

Blcyclo[2.2.1]heptane.2.2.5.6 tetrachloro
1,7-bts(chloroMthyl) -7-(dtchloroaethyl) -
Btcyclo|2.2.1)heptane.2.5.6-trlchloro-3.
3-bls(chloroaethyl)-2-(dlchloroM>thyl)
(exo.exo.exo)-

Blcyclo)2.2.1]heptane.2.2.5.6-tetrachloro-
1.7-bts(chloroaethyl 7 (dlchloro«ethyl)-
(5-endo.6-exo.7-antl)-

Blcyclol2.2.1]heptane.2.2.5.6 tetrachloro
1.7-bls(chlor«Mthyl-7-(dlchlorowtbyl)-
(5-endo.6-exo.7-syn)-

Blcyclo[2.2.1Jheptane.2.3.5.6 tetrachloro-
7-(chloroMthyl)-l-7-bl$(dtchlor(MKthyl)-
(2-endo.3-exo.5-endo.6-exo.7-syn)-

Blcyclo[2.2.1Jheptane.2.3.3.5.6-pentachloro-
2.2.S endo.6 exo.8.9.10
heptachlorobornane;
loxaphene toxicant B
loxaphene toxicant C*;
2-endo.3.3.5.6-exo.8.9.
10.10-nonachlorobornane

Toxic fraction A;  2.2.
5-endo.6-exo.8.9.9.10-
octachlorobornane

2.5.6-6X0.8.8.9.10-
heptachlorodlhydro-
caophene

2.2.5 endo.6 exo.8.8.9.10
octachlorobornane;
loxaphene toxicant A-l

2.2.S endo.6-exo.8.9.9.10
octachlorobornane;
Toxaphene toxicant A-2

Toxaphene toxicant Ac
loxaphene toxicant  C4;
?,3,3-endo:5:6-exo,a,9
10.10-nonachlorobornane
Clark and Natsuwira. 1979;
Saleh and Caslda. 1978;
Saleh et al.. 1979;
Chandurkar and NatsuMira.
1979a.b

Chandurkar and Matsuaura.
1979a.b
Clark and HatsuMira. 1979
Swanson et al.. 1978
Pollock and Kllgoie.  1980
Pollock and Ktltjore.  1980
Chandurkar et al..  1978
Clark and NalsuMira.  1979
         •Proposed structures have not been confirmed                            ,

          CAS RN = Chemical Abstracts Service Registry Nunber; Nf -- Holecular Fornula; 9C1 -- Ninth Collective Index.  Chealtal  Abstracts
O
CJ

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toxaphene  Is  a  complex  mixture and the quality of  the  commercial  product  Is
so  variable.   Thus,  the  lower  chlorinated components  will  be  more  water
soluble, and  more  volatile than those with a higher  chlorine  content.   Odor
detectlbUHy by  one  Individual  of  toxaphene  dissolved In  water has  been
found to be 0.14 mg/i at 60°C (Slgworth,  1965).

    Selber  (1982)  noted  the  occurrence  of   differential   volatilization.
Toxaphene  Is  mlsdble  In  acetone, benzene,  carbon  tetrachlorIde,  ethylene
dlchlorlde,  toluene  and xylene;  It  Is soluble  (g/100  mt at  27°C)   In  tur-
pentine  (350-400),  kerosene  (>280),  and  fuel  oil   (250-275)   (Brooks,  1974).
Solubilities  In alcohols such as  Isopropanol (15-18)  and  95X ethanol  (10-13)
are generally lower.  The  heat  of  fusion  1s  0.39 cal/g;  the  specific  heat  Is
0.258 cal/g/°C at  41°C;  the  viscosity Is 1.4 poise at  100'C (Brooks,  1974).
The octanol/water coefficient was  variously found to  be  -3300  (Paris  et al.,
1977) and  825 (Sanborn  et al.,  1976).   The  diffusion  coefficient of  toxa-
phene  1n  water  at  15°C  has   been   estimated  to  be  5.6x10"*  cm2  sec"1
(Velth and  Lee,  1971).   The  KQC  value at 25°C  Is  98,600 (McDowell   et  al.,
1981).  The physical properties of these  products are  provided In Table II-3.

    Toxaphene  dehydochloMnates  when heated  above  120°C,  or  exposed  to
ultraviolet light (mercury arc) or Intense sunlight  (Brooks,  1974).

    Toxaphene (20 mg  of  a  90%  toxaphene/lOX xylene liquid)  when  placed In  a
sealed  tube  for  15  minutes will  produce different  IR  spectra at  250°C;
volatilization occurs above  125°C  (Kennedy et al.,  1978).  When  heated In  an
open  crucible  In  a  muffle   furnace   for  45 minutes,  the  toxaphene/xylene
02000                                11-5                            02/25/87

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o
g TABLE 11-3
o
Physical Properties of the Degradation Products of Toxicant Bd
CAS RN Purity mp Retention Ttmeb *»N1-EC Response Molecular
(X) (°C) (minutes) Relative to Hlrexc Formula
57981-29-0 99 155-156 14.00 23 C10H12C16
64618-63-9 99 107-108 11.15 23 C10H10C16

7 65620-64-6 90 146-148 13.23 13 CloHl2n6
o>
66157-70-8 99 100-101 8.45 11 ClOHllc15


Diagnostic
CI/NS Peaks
M-C1. M-HC1.
M-C1-HC1
HO. H-C1,
H-HC1
H-C1-HC1
H-C1. M-HC1,
M-C1-HC1
M*l. M-C1.
H-HC1
    aSource:  Saleh and Caslda. 1978
    bThe retention time of  Nlrex on an open  tubular  column at 200°C was 45.50 minutes; the retention  time  of
     toxicant B was 20.41  minutes, and Its relative EC-response was 51.
    CEC = Electron Capture;  Nlrex response Is taken to be 100.
    CAS RN -  Chemical Abstracts Service Registry Number
o
CO
CD

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mixture showed a weight  loss  of  94.2X at  400°C  and 99.9% at 1000°C.   Notice-
ably  different  gas  chromatograms were  noted  at 300°C for  the  open  crucible
experiments.  At  400°C,  the major gaseous products were  tentatively  Identi-
fied  as hydrochloric  add,  chlorine,  phosgene,  carbon tetrachlorlde,  chlori-
nated allphatlcs containing two  carbons,  carbon dioxide,  carbon monoxide  and
water.   In  catalytic  hydrodechlorlnatlon, the gemlnal   chlorines  are more
readily attacked than  the other  chlorines  (Kranlch  et al.,  1978).   Thus,  <2%
of  toxaphene  degradation  products  containing  more  than  five   chlorines
remained  after  19  hours of  reaction  (at  100'C,  50 bar  pressure);  a 10 g
toxaphene  1n ethanol  per gram  N1  on  Kleselguhr  was used  as a  catalyst.
Toxicants  A  and  B  appear to be  stable  under  the  conditions  of  gas  chromato-
graphy (Khalifa et al., 1974).

    DehydrochloMnatlon reactions are accelerated  In  the  presence  of  a-lkalls
and  Iron  compounds  (Brooks,  1974),  that   1s, when toxaphene Is refluxed  In
alcoholic  potassium  hydroxide (Archer  and  Crosby,  1966; Gomes, 1977;  Crist
et al., 1980),  or  when toxaphene 1s  In the presence  of  hematIn/sodium  thlo-
sulfate (Khalifa et al.,  1976).

    The colorlmetMc  technique  formerly  used depended upon  the reaction  of
toxaphene  with  dlphenylamlne  and zinc  chloride at  205°C to give a  chromo-
phore with ax    of 640 nm (Graupner  and  Dunn.  1960).
              max

    Studies  on  Toxicant  B were  reported  by Saleh  and Caslda (1978).   Toxi-
cant  B undergoes  reductive  dechlorlnatlon at  the germinal  dlchloro  group  to
yield   2-endo,5-endo,6-exo,8,9,10-hexachlorobornane   and    2-exo,5-endo,6-
exo,8,9,10-hexachlorobornane 1n many systems.


02000                                II-7                             02/25/87

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Production and Usage
    Toxaphene  production   In  the United  States was  18.4 Mkg  In 1978,  and
accounted  for  more  than  13%  of  total cyclic  pesticide production.   Three
manufacturers who  reported their production or  sales  figures for that  year
were  Tenneco Chemicals,  Inc.,  Hercules,   Inc.  and  Vertac,  Inc. Vlcksburg
Plant  (USITC, 1979).   The  United States toxaphene consumption for the  years
1978,  1979 and 1980  was 15.5,  13.5  and 13  Mkg,  respectively  (FAO, 1982).   In
1980,  -1  Mkg  was applied  to  soybean  acreage.   This  1980 usage  represented
23.8%  of  the  total  Insecticides  applied to this crop  (Elchers and SerleUs,
1982).  During  fiscal  year 1980  toxaphene  was  also used by  the  U.S.  Forest
Service for  tick and  lice control   on  3030 cattle.  A  total of  1607  acres
(650 ha) were Involved In  this process (USOA,  1981).

    Primary utilization occurs 1n agricultural  crop  applications, especially
cotton  (IUPAC,  1979).   It  1s  also  extensively  used for  the  control  of  exo-
parasltes   on  livestock.   The  application  rates   vary   from 1.2-9.6  kg/m3
water  (IUPAC,  1979).   The  cumulative  world  usage  of toxaphene from  1946  to
1974 was -450,000 tons {IUPAC, 1979).

    Technical grade  toxaphene  can be supplied as a  40%  dust  concentrate  and
as a  viscous liquid  containing  10%  xylene  (Brooks,  1974),   with  the  liquid
being  vulnerable  to  degradation  by  metals,   such  as   Iron  and aluminum.
Emulslf1able concentrates  containing 4,  6  or 8  Ib.  of  toxaphene/U.S.  gallon
(0.48,  0.72  or  0.96  kg/i)  are   formulated   usually   with  xylene,   0.5%
eplchlorohydrln  and  an appropriate  emulslfler  (1f water  Is the carrier),
Uettable  powders  (40%  toxaphene  content),  dusts (5-20%)  and  granules  (5  and
20%)  are  also  used  (Brooks,  1974;  IUPAC, 1979).   Toxaphene has also  been

02000                                11-8                            02/25/87

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coformulated  with  other   pesticides  [e.g.  DOT,  Maneb,  Z1neb,  dioxathlon,
dlazlnon,  malathlon,  methylparathlon,  endrln,  endosulfan,  Isobornyl  thlo-
cyanoacetate  plus related  toxaphene terpenolds  (ThanH'e),  pyrethrlns,  and
plperonyl  butoxlde]  (Brooks,   1974).    A   selected   list  of  recently  used
coformulatlons  Is  provided 1n  Table II-4.   In  addition  to such  uses  toxa-
phene  has  been used  as  a protective agent  to reduce phytotoxlc  effects  of
such  other  pesticides  as phosphorothlolates  (Aller and  Hansen,  1981),  to
provide broad  spectrum pestlddal  properties  In  combination  with  such herbi-
cides  as  benazolln (Anonymous,  1982),  and to  produce  more potent  1nsect1-
ddal mixtures. I.e. with AmHraz (Kerry and Welghton, 1979).

Spectroscoplc Properties
    Toxaphene  components  are  not aromatic and,  therefore, would  be  expected
to absorb light at wavelengths below 250 nm.

    Infrared  analysis  1s   still  used for  qualitative  Identification.   There
are  prominent  absorption bands  1n  the  6-8 ym  region  and  the  specified
absorbence at  7.2 vin of  0.0177  (maximum)  1s utilized (Brooks,  1974).   The
Infrared  spectrum of toxaphene was  reported by  Archer  and Crosby  (1966).
Since  toxaphene Is  a  complex  mixture, mass spectroscopy  (MS)  of  the mixture
Is  not  diagnostic.   However,  upon  gas  chromatography/mass  spectroscopy
(GC/MS), the  presence of  the  most  toxic components  can be  found  1f  adequate
resolution can  be  obtained on the  gas chromatographlc column.   The  electron
Impact  and  chemical  1on1zat1on mass  spectra  of Toxicant  A  and B  (see Table
II-2)  were  recorded  1n  Holmstead  et al.  (1974).  Because  the  compounds  are
02000                                11-9                            02/25/87

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                                                                           1ABLE II 4

                                                        Cheated! Information Regarding loxaphene Mixtures
          CAS RN
                                 Molecular forMula
                                                                             ChcMtcal Na«es
                                                                                                                                 SynonyMS
o
u>
OD
        8073-68-5


        8077-22-3





        37271-99-1


        37272-06-3
39295-90-4


39295-93-7



39295-94-8


39400-89-0


39474-75 4



50641-56-0


55267-24-8


58386-60 0
                       C4HgN202S-Unknown
                       C|4H9C1.,.CHH,UIIOSPS
                       Unknown
                       C8HiQNOsPS-Unknown
                                Unknown
                               Unknown
                               C)4HMM04PS.Unknown
                                          .C5H,2M03
                                PS2-Unknown
                                C)oH)4MOi,PS. Unknown
                                c16H15cl3°2*Unknown
                                Cj0H)906PS2-Unknown
HhanlMldothlotc actd.N ((d*lnocdrbonyl)oxy)-.
•ethyl ester. «»xt. with toxaphene (9C1)

Phosphorothlotc acld.O.O-
-------
                                                                        1AHI t  11 -4 (cunl.)
rsj
           CAS HN
  Molecular foraula
                                                                                      Chemical Nanes
                                                                                                          Synonym
 O
 CJ
 CO
         58703-96-1




         58934-38-6



         64034-61-3

         67872-25-7




         70028-45 4


         70162-18-4



         73755-37 0




         7/5I8-58-?



         77518-59-3



         77837-51-5



         77837-52 6
C,4H,Cl5.C,0Hi4IIOs
PS-Unknown
CH4M20-Unknown
C8H|oMOsPS-Unknown
C4H7Br2Cl204P-
Unknown

         Unknown
«0SPS.Unknown
c25H22c1N03*Unknown
Phosphorothlulc actd.O.O-dtethyl-0-(4-n1tro-
phenyl)ester. •III. with toxaphene and 1.1'-
(2.?.^-lrlcbloroethylldene)bls(4-chloroben2ene)
(9C1)

Cyclohexane.l.2.3.4.5.6-hexachloro-.(l alpha.
2 alpha. 3 beta. 4 alpha. 5 alpha. 6 beta)-.
•Ixt. with toxaphene (9CI)

Urea. alxt. with toxaphene (9CI)

Phosphorothlolc acid. 0.0 dlethyl 0 (3.5.6
trtchloro 2 pyrtdtnyl)eiter. atxt. with 0.0
dimethyl-0-(4-nltrophenyl)phosphorothloate
and toxaphene (9C1)

Phosphoric acld.l.2-dlbroMo-2.2-dtchloroethyl
dl«ethyl ester. *lxt. with toxaphene (9C1)

Nethanl«)da«tde. M' <2.4 dl«ethylphenyl) M
(112.4 -dlaethylpheny 1) l>lno)aethyl) -N-Mthyl -.
•lit. with toxaphene (9C1)

Phosphorothlolc acid. 0.0 dlaethyl 0 (4-nltro-
phenyl)ester. alxt. with M' (4 chloro 2-
•ethylphenyH-M.N-dtaethylaethanlMtdaatde
and toxaphene (9CI)

Phosphorothlolc acid. 0-ethyl-S-propy1-0-
(2.4.6-trlchlorophenyl)ester. «lxt. with
toxaphene (9CI)

Phosphorothtolc acid. 0-|4 bro«o 2-chloro-
phenyl)-0-ethyl-S-propyl ester, alxt.  with
toxaphene (9CI)

Ben/enedcetIc acId.4 -chloro-aIpha-(1 -
•ethylethyl)-.cyano(3-phenoxyphenyl)iMthyl
ester. «lxt.  with toxaphene (9C1)

Cyc lopropanerar boxy lie acld.3-(2.2-dlchloro-
etheny1)-2.2 dlaelhy1 -.(3-phenoxypheny1)
•ethyl ester, ntxt. with toxaphene (9C1)
Hexatox
                                                                                                     TXL
loxaphene-naled
•txture
         CAS RN =- Che«lcdl Abstracts Service Registry Number; 9CI -- Ninth Collective Index. Chenlca)  Abstracts

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allcycllc,  the mass  electron  Impact  spectra  have  extremely  weak or  non-
existent  parent  Ion  peaks,  the  major  Intense  peaks  being  In  the low  m/e
range.   Thus,  chemical  1on1zat1on MS after  GC  1s more  useful  than electron
Impact MS (Holmstead et al., 1974).

    Although  capillary  GC  showed Toxicant A  was  one peak,  90 and 270  MHz
'H-NMR   spectroscopy   revealed  1t  to  consist  of   two   major   components
(Matsumura  et  al.,  1975).   The  structure of  Toxicant  Ac  was deduced  pri-
marily through  Its 'H-NMR spectra  (Chandurkar  et  al.,  1978),  since  Us  gas
chromatographlc properties were similar to those of components A-l  and  A-2.

    Saleh and  Caslda  (1978)  recorded  the chemical   1on1zat1on/mass  spectra
(CI/MS)  of  the major In  vitro  and U> vivo degradation products of  Toxicant B
(see  Chapter  III).  The chemical  1on1zat1on/mass  spectrometrlc  (CI-MS)  data
can be found In Table III-7.

    The  'H-NMR  spectra  of  Toxicant  C  and  Us  major metabolite  (CR  70459-
31-3) (Table III-7) was reported by Chandurkar and Matsumura (1979b).

Analysis
    The  major  recent  analytical  methods  for  toxaphene  In  a  variety  of
matrices are  summarized  In Table  II-5.   Most of the  methods  employ  extrac-
tion  from   the  suitably  prepared  substrate,  a  Florlsll  clean-up  step,  and
electron  capture/gas  chromatography  (EC/GC),   or  GC/MS.   In the case  of
tissue analysis,  a silica  gel clean-up  step  1s also often  utilized  before
EC/GC or GC/MS (Haseltlne  et  al.,  1980).   A  nonpolar phase  like  OV-101  Is
generally utilized for gas chromatography.

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                 1ABLE  II 5




ExaMples of Analytical  Methods for Toxaphene
o
0
o
















l~i
1
CJ















G»
00
Substrate

PrlMry sludge
(20 g)




Duck tissues
(brain, liver.
blood, eggs and
carcasses)
Soil (1 g)


Bovine deftbrtnated
whole blood
circulated through
a per fust on
apparatus (2 at)






HI Ik (20 M)


Butter (1 g fat)


Heat (10 g)




Method

Extraction; concentration; florlsll coluwi cleanup; EC/GC





Extraction; florlstl coluwi cleanup; silicic acid separa-
tion; EC/GC (Gel peraeatton chroaatography for blood and
young carcasses)

Florlsll extraction and cleanup;
Dehydrochlorlnatlon with KOH;
GC
I. Agitation with H?0 and hexane;
centrifuge; hexane extraction;
GC

II. Agitation with 88X f orate acid
and hexane; solvent phase shaken
with I^COj; hexane extraction; GC
HI. Agitation with 88X foralc acid;
florlsll extraction and cleanup;
GC

5X Dlethyl ether -hexane elutton; GPC solvent Isolation;
acid oxidation; hexane extraction; florlsll coluwi
cleanup; EC/GC
5X Olethyl ether -hexane elutlon; GPC solvent Isolation:
acid oxidation; hexane extraction; florlsll coluwi
cleanup; EC/GC
5X Dtethyl ether -hexane elutlon; GPC solvent Isolation;
acid oxidation; hexane extraction; florlstl coluwi
cleanup; EC/GC


Concentration Recoveries Reference
Added
50 Mg 92»27X Rodrlquei et al.. 1982
84.2X
91»7X
15 t,g 89»28X
89»19X
93»22X
10-20 ppa 113X Haselttne et al.. 1980
(average) :


1.0 Mg 84. 9X Crist et al.. 1980
0.5 1,9 71. 4X
0.1 M9 71. IX
20 tig/at 73. 4X Nalortno et al.. 1980



20 Mg/*t 71.7X


20 Mg/*i 103.4X

0.3-15 Mg/at 96. 8-102. IX
(range)
0.5-5.0 ppa 78-88X Boshoff and Pretortus.
(range) 1979

0.5-5.0 ppa 79-86X
(range)

0.5 5.0 ppa 76-79X
(range)




-------
    Since  toxaphene  1s  a  complex  mixture,  the  chromatographlc  pattern  of
standards may  not match  that  of  the residual  toxaphene remaining In environ-
mental  samples,  since differential  volatilization,  sorptlon,  solub1T1zat1on
or  mlcroblal  degradation  may  have  occurred  that  changed   the  diagnostic
pattern.   However,   certain  components  of  toxaphene  are more toxic  than
others; these  components  can  be  selectively quantHated  by the methods given
In  Table  II-.5.   GC/MS   techniques  are  Invariably  the  ultimate  forms  of
analysis  after a EC/GC  screening  step,  with  chemical  lonlzatlon MS  being
preferred since electron Impact MS produces no or very weak molecular  Ions.

    Some of  the  older methods for detecting  toxaphene  such as total  organic
chloride, coloMmetrlc  methods,  or  bloassays  using  Insects,  were  generally
nonspecific.   Since  toxaphene Is  an extremely complex mixture,  and not very
heat  stable,  the question  of thermal  degradation  on  GC columns  remains  a
vexing  one.   Although  there  are  three  major chromatographlc  peaks  (Gomes,
19.77),  the  use  of  a flash  heater  filled with  various reagents In the Injec-
tion  port  produced  substantial  changes  1n   the  chromatogram  (Mlnyard  and
Jackson,  1965).   Treatment of  toxaphene  residues  with  alcoholic  potassium
hydroxide  followed   by   column  chromatography  (detection  to  1  ng)  causes
dehydrohalogenatlon with  the  appearance  of  one large single peak (Archer and
Crosby, 1966;  Gomes, 1977; Crist  et al.,  1980).   Recoveries  from soil  are
claimed to  average  between  71 and 85X at  levels  from 0.1-1 ppm toxaphene In
soil, although the method did not  appear to be useful for rat adipose tissue
(Crist et al., 1980).
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    Reviews  by  Selber  (1982) and  Pollock  and KHgore  (1978a)  highlight  the
difficulties  of separating  toxaphene  from  other chlorinated  hydrocarbons,
and pesticides.

    Analysis  for  toxaphene  metabolites has  Involved  extraction  techniques
followed  by  thin-layer  chromatography  (TLC)   (Chandurkar  and  Matsumura,
1979a,b).   Pollock   and   Kllgore   (1980)  used  TLC   to  circumvent  artifacts
caused by thermal degradation on GC columns.

Summary
    Toxaphene  Is  a  mixture  (average  molecular  weight  414) of at  least  177
compounds,  mostly  chlorinated  camphenes,  which  has been  used  as  a  broad
spectrum  contact  pesticide  (e.g., In  soybean  and  cotton  fields,  and  for
ectoparasites).  It Is produced Industrially  by the  reaction  of camphene  and
chlorine  (In carbon tetrachloMde solvent)   1n  the  presence  of  ultraviolet
light to  produce a  product  of highly variable  quality.   Toxaphene can  exist
as a  40% dust  concentrate,  a 9:1   liquid mixture  containing 10% xylene  or  as
emulslflable concentrates.   It  1s  extensively coformulated  with other  pesti-
cides (e.g., DDT, parathlon, endrln).

    The  vapor  pressure,  density,  solubility and  other  physical  properties
vary according  to the  quality of   the product.  Toxaphene Is  nonpolar  and  so
Is mlsdble  In nonpolar  solvents, but less  soluble In  polar  solvents.   It
adsorbs  avidly  to  soils  and partitions readily  Into n-octanol.   Toxaphene,
being aliphatic,  Is  a  poor  absorber  of ultraviolet  light  above wavelengths
of 250 nm.   Gas chromatography with electron capture or  chemical 1on1zat1on
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mass  spectrometMc  detection  are the most commonly  used  analytical  methods.
'H- and  13C-nuclear magnetic  resonance  techniques  may  be  used  to  assess
the "fingerprints" of toxaphene fractions.

    Toxaphene readily  dehydrochloMnates  when heated above 120°C, but  Toxi-
cants  A  and  B  appeared stable  In  gas chroroatography.   Dehydrochlorlnatlon
also  occurs  In  the  presence  of  alkali  (used as  an analytical method),  of
reduced hematln,  and of Iron  compounds.  Reductive  dechlorInatlon occurs  on
exposure to  ultraviolet light In the  radiation  range (230-290 nm)  of  toxa-
phene In organic solvents,  preferentially  at  the  C« and  C? positions.

    Most of  the analytical techniques  to detect  toxaphene metabolites  have
utilized thin-layer  chromatography  as  the end  step.  Toxaphene  In  environ-
mental samples  and  tissues often does  not  have  the  chromatographlc  pattern
of  a  toxaphene  standard  so  that  recognition by  gas chromatography  occurs
primarily by  retention  time,  and quantHatlon (detection  to  1  ng)  Is  gener-
ally  accomplished by the method  of  alkaline  dehydrochlorlnatlon,  extraction
and  chromatography   (Gomes,  1977).   Metabolites  are generally Isolated  by
thin-layer  chromatography or liquid  chromatography.
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                             III.  TOXICOKINETICS

    This  chapter  refers  only  to data relevant  to  warm-blooded vertebrates.
Residue estimation  In  tissues  after  toxaphene exposure 1s generally based on
the  comparison  of  EC/GC  traces  of a test  sample  with the  pattern  of stan-
dards.  Systematic  errors  In residue  levels  may exist because of the differ-
ential behavior  of the >177 components  of  toxaphene  relative  to  metabolism
can render such comparisons Inappropriate.

Absorption
    Toxaphene  1s   absorbed  through  the  skin  (especially  1f  mixed  with
xylene),  the  lung,  and the gut  (IUPAC,  1979;  IARC,  1979).   The  rates  of
absorption have  not been  documented.  The available data are  summarized  In
Table III-l.

Oral
    One  Incident   that  Illustrates  toxaphene absorption  Involved  toxaphene
poisoning  of  the  Tule Lake Basin of Northeastern California  and  Southern
Oregon (Keith, 1965).   Toxaphene,  -2.2 kg/ha, was  used between 1956 and 1963
In areas  on and  off the Tule  Lake and Lower Klamath Refuges,  with heavy use
occurring  between  1957  and  1960  (>1000 ha  treated/year)   In  Modoc  County.
Though  many   pesticides.   Including  DOT,  were also  used 1n  the  area,  the
wildlife  bird mortalities coincided  with the significant use  of  toxaphene.
Toxaphene was found 1n the carcasses and  tissues  of dead birds In 1960-1961
(0.3-15  mg/kg  whole  carcasses  or   organ  composites)   (see   Chapter V  for
discussion on residue  levels  as  linked to  mortality).   Keith (1965)  also
reported  on the mortality  of other birds (Robinson, 1959) In Nebraska and In
Montana upland that were associated with toxaphene application.
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                                 TABLE  III-l

                 Absorption Data for All Routes  for  Toxaphene
Absorption
Route
Oral




Species
birds
birds
bald
eagle
birds
range
birds
Exposure
2.2 kg/ha
unknown
unknown
unkown
unknown
Effect
kills In N.E. California
and southern Oregon;
residues In carcasses
kills 1n Nebraska
and Montana; residues
In carcasses
death; residues 1n
carcasses
kill; residues In
carcasses
kill; residues In
carcasses
Reference
Keith, 1965
Robinson, 1959
Barbehenn and
Relchel, 1981
Pollock and
Kllgore, 1978a
HcEwen et al . ,
1972
             birds



             raccoon


             horse


             pig
unknown



unknown


unkown


unknown
kills 1n California,
South Dakota, Texas
and Arizona

kill; residues In
carcass

poisoning, residues
poisoning; residues
             cattle
various
poisoning; residues
In tissues, milk
U.S. EPA, 1977
U.S. EPA, 1977
Mount and
Oehme, 1981

Buck et al.,
1976

Mount et al.,
1980

D1P1etro and
Hallburton,
1979

Claborn et
al., 1963

Zwelg et al.,
1963
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            III-2
                                02/25/87

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                             TABLE III-l  (cont.)
Absorption
  Route      Species    Exposure
                               Effect
                                         Reference
Oral (cont.) cattle
           various
Oral
Inhalation
Dermal
humans
             humans
animals

humans
humans


animals



humans



pig



animals
unknown
           unknown
various

unknown
                        various
                        various
unknown
(Undane/
toxaphene)

various
                        various
             poisoning;  residues  In
             tissues,  milk
three fatal; two
poisoning
poisoning from
contaminated fish;
8 fatalities;
44 poisonings

1059 values

bilateral hllar adeno-
pathy with fine mlHary
lesions on lungs; acute
pneumonia; blood
eos1nophH1a; serum
globulins Increased

no toxaphene residues
(<0.1 mg/kg blood)

L050
Illness
                                     nervous  system
                                     disorder
             1050 values
Steele et al.,
1978

Steele and
Mount, 1980

Steele et
al., 1980

McGee et al.,
1952

U.S. EPA, 1977

U.S. EPA,
1977,1980
see Table V-l

Warrakl, 1963
                                       U.S.  EPA,  1980
Boots Hercules
Agrochemlcals,
Inc., n.d.

Pollock, 1958
                          D1P1etro and
                          Hallburton,
                          1979

                          see Table V-2
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    Toxaphene residues  have  been  measured 1n the  tissues  of  dead  wild  birds
(U.S. EPA,  1979a).   The carcass/brain residue ratio  In bald  eagles  was  6.7,
similar  to  that  of  ds-nonachlor,  PCBs,  oxychlordane and ds-chlordane  In
the same  species  (Barbehenn  and Relchel, 1981).   The  ubiquity  of  the occur-
rence  of  toxaphene   residues   In  birds  Implies   that toxaphene  absorption
occurred by the oral  and  possibly percutaneous  routes.  In fact,  Pollock and
KHgore  (1978a)  attributed two bird  kills  to  eating of  contaminated  fish,
and direct  toxaphene  exposure.  McEwen  et  al.  (1972)  noted no change In  bird
numbers 1 week after  spraying  toxaphene on a range  land,  but during the 2nd
week  a  significant decrease  In bird numbers  was  observed and several  bird
carcasses contained 0.1-9.6 mg  toxaphene/kg  ww.  Live birds  In the  area had
carcass residues  of  around 0.4-1.0  mg/kg ww.   Other  cases  of bird kills are
summarized  1n Table III-l.

    Poisoning Incidents  In domestic animals-also  demonstrate  that  absorption
can occur  by  the  oral  route  (see Table  III-l).   The U.S. EPA (1977)  docu-
mented  38  specific cases  of toxaphene exposure  to  wildlife and  livestock
between 1966  and  1975,  Including 10  for cattle,  3  for  swine, 2  for  horses
and 1  for  sheep.   Animal oral  L05Q values  In toxlclty  experiments  depend
on  the  vehicle;   I.e..  readily  absorbed  oils  like corn  oil  and   peanut  oil
elicit  low  LD5Q  values,  whereas  carriers  not readily  absorbed   like  kero-
sene yield higher LD5Q values (U.S.  EPA, 1980) (see Chapter V).

    Evidence  for  oral absorption  In  humans  resulting  In  toxaphene poisoning
1s summarized 1n Table III-l  and treated fully 1n Chapter VI.
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Inhalation
    The  relevant  evidence for absorption  by  the Inhalation route  Is  summa-
rized 1n Table IH-l.

Dermal
    Data In  animal  experiments  (IUPAC,  1979)  Indicate that  toxaphene  can  be
absorbed through  the skin.  However, the  toxldty  1s generally an  order  of
magnitude  less  than  acute oral  ID™  values  using  the  same  carrier.   The
LD5Q  values  are  also  very  vehicle-dependent  (see  Chapter V).   The  values
for the  rabbit  range from <250  mg/kg for toxaphene  applied 1n  solution  to
<10QO-2000 mg/kg for toxaphene applied as a dust.

    01P1etro and  HalVburton  (1979)  reported an  episode of  toxaphene toxico-
sis In  swine after  topical application  36 hours earlier  with -10  times  the
prescribed  toxaphene  dose  (300  mJU  of  61X  toxaphene   stock   In  4 i   of
H_0) for sarcoptlc mange.

Distribution
    Since there are  over 177  components of toxaphene  that  do  not  metabolize
In the same manner,  nor  at the  same  rate;  their Identification can be diffi-
cult as  a result  of the  change  of   the  diagnostic  EC/GC  chromatogram  from
that of  a  reference  chromatogram.   This  problem has been observed  1n  milk
analysis (Cairns  at  al.,  1981) and  analysis  of bird organs  (Haseltlnc  et
al., 1980).  One  solution 1s  to dechloMnate the remaining toxaphene.   This
type of  analysis  has been performed  by Chandurkar and Matsumura (1979b)  for
Toxicants B  and C  In rats and by Saleh  and Caslda  (1978)  for Toxicant B  In
rats.    Another  method  1s  to study  the metabolism  of only  the more  toxic
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components   of   toxaphene.    This   has   been  performed  by  Chandurkar  and
Matsumura  (1979b), and  Saleh  and  Caslda  (1978).   A  third  approach  Is  to
utilize  radlolsotopes.   This   has  been  done  by  Bush  et  al.   (1978)  using
broiler chickens,  by  Chandurkar and  Matsumura  (1979a),  Ohsawa et al.  (1975),
and  Crowder  and  Dlndal  (1974)  1n rats.   However,  chromatographlc  confirma-
tion  Is  still  necessary  for  the labeling experiments  since  the position  of
the  label defines  the significance  of  the results.   Unfortunately,  no single
study has compared all methods  simultaneously.

     In general,  toxaphene appears  to be  rapidly metabolized  and Us  metabo-
lites quickly excreted In most species,  with  fat  as  the  preferred  tissue  of
storage (IUPAC,  1979).

Distribution 1n Animal Tissues
    Birds.   A  number  of  studies   In   avlan  species  such  as  ring-necked
pheasants (Genelly and Rudd,  1956a)  and White  Leghorn chickens  (Bush  et al.,
1977, 1978), suggest  that  birds can  metabolize toxaphene  adequately,  as long
as the doses  are not   too high  and  they can  tolerate  steady Ingestlon of the
chemical   for  a  period of  months  without serious effects at  the lower  doses
(Genelly and Rudd,  1956b; Page et al.,  1978), with the exception of  altered
cartilaginous  structures  (Page et  al.,  1978).    Young  white   pelicans  and
young  water-fowl  are  more  susceptible  to  toxaphene  (Keith,  1966)  (see
Chapter V).

    Concentrations of 0, 25, 100,  200  and 300 ppm toxaphene 1n  the  form  of
dietary mash  were fed to  adult ring-necked pheasants  (10/sex)  for up  to  94
days  (Genelly and Rudd,  1956a).  The  mean doses for 100 and 300 ppm levels


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were  4.74 and  9.12 mg  toxaphene/blrd/day,  respectively.   Organic  chlorine
residues  were  found 1n fat for all  doses  >4.74  mg/b1rd/day,  and 1n  liver at
the  9.12  mg/b1rd/day  dose.   One  bird  at  the  4.74 mg/b1rd/day  dose  showed
high residues  In the testes.

    Toxaphene  at 0,  0.5,  5, 50 or 100 ppm diet  was  fed  to  five groups of 90
1-day-old  female White Leghorn chickens  per  group for up  to  34 weeks (Bush
et  al.,  1977).  The  pesticide,  as  Tox-SOl-6,  containing  59%  active  Ingre-
dient, was  first dissolved  Into  corn oil and  then Into a formulated diet.
The  level of  toxaphene  In  excisable  adipose  tissue  of  8-week-old  birds
Increased with Increasing dietary toxaphene as described by the following:

             mg toxaphene/kg body  fat =  3.4 F  +  33.5    r2 = 0.725

where F 1s the level of toxaphene In the  feed  In  ppm.  This amount  accounts
for only  50% of the  total  toxaphene Ingested.  Accumulation  In 32-week-old
birds was less than  In  8-week-old  birds  despite  continued exposure.   Bush et
al. (1979) suggested  that  White Leghorns were able  to metabolize or excrete
toxaphene at  a rate  comparable  with dally  Intake.   Residues  were  found In
the eggs of bird receiving dietary levels >5 ppm.

    Toxaphene-»*Cl   was  utilized  In  a study  by Bush et al.  (1978)  to  assess
the distribution  of the  labeled  pesticide In broiler chickens.  Concentra-
tions of  0,  0.22,   0.40,  2.16  and 3.82  ppm  toxaphene-3*Cl  were administered
to  1-day-old  unsexed  broiler  chicks (three  replicates  of 30  birds  each).
Feed  and  water were   supplied  ad  libitum  under  a   regime   of  continuous
Incandescent  light.

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    For  toxaphene  (T)  In  tissue (mg/kg ww)  versus toxaphene  In  feed  {ppm)
after  8  weeks of  continuous  feeding  (F),  the following  relationships  were
found:

    Adipose:                  T = 4.081  F + 0.765            r2 = 0.96
    Gizzard:                  T = 0.1588 F * 0.0140           r2 = 0.7484
    Heart:                    T = 0.1659 F * 0.0565           r2 = 0.9305
    Kidney:                   T = 0.0942 F *• 0.0400           r2 = 0.7512
    Liver:                    T = 0.0355 F + 0.0625           r2 = 0.4528
    Breast Muscle:             T = 0.0472 F > 0.0018           r2 = 0.6269
    Leg Muscle:               T = 0.1307 F * 0.0241           r2 = 0.8669

    The Intercepts  In these equations are probably not different from zero.

    Only  In  adipose  tissue  did appreciable  amounts  of toxaphene  accumu-
late, but this still represented only 5% of the consumed toxaphene.

    One-year-old black  ducks  of  Anas rubMpes were  fed  dietary levels  of 0,
10 or 50  ppm  toxaphene  with  the  toxaphene dissolved In a volume of propylene
glycol equivalent  to  IX of  the  total diet (Haseltlne et  al.,  1980).   After
hatching  of  eggs,  the  residues  1n  ducklings  fed  on the  same  diets  as  their
mothers were  monitored  at  days  14 and  84  post-hatching.  At day 14,  the
ducklings contained  6.3 and  20.8 mg toxaphene/kg bw  for  the   10  and  50 ppm
dietary  levels,  respectively.   At  day  84,  female  ducks  contained 7.7  and
28.0 mg/kg  carcass.   GC chromatograms  of  body residues were  similar  to the
toxaphene standards.
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    Haseltlne  et  al.  (1980)  also  performed  a  one-generation  study  on  the
distribution  of  toxaphen_fi  In black  ducks  (15  pairs/group).   Toxaphene  was
administered  to  newly  hatched  ducks  (F.  generation)  continuously  over  19
months 1n propylene  glycol  admixed  so that  the carrier constituted IX of the
duck  mash  feed.   All  ducklings obtained  from  the Fl   and  Fl   breedings
                                                        3D
were  fed  the  toxaphene  containing diets until  12  weeks  of age.  The techni-
cal  toxaphene  concentrations   1n  the diet were  0,  10 or  50  ppm.   Toxaphene
residues  1n  liver,  brain,  blood  and carcass  were  quantified  by  direct  com-
parison of  the total area  under  the GC  curve with  the  total  area  under  the
standard  toxaphene  curve (Table  III-2)  or by  comparing  the area  under..two
characteristic peaks  with  the area  under  the  same peaks 1n  the  standard.
The  two  methods  did  not agree,  Indicating  that differential  metabolism of
the toxaphene  components  had  occurred.  Liver  residues  for  the 50 ppm group
at month  19 constituted  only 12%  of  the cumulative  dietary  levels.  Brain
residues were  higher  \n  males  (1-3  mg/kg ww)  than 1n females (-0.5 mg/kg ww)
exposed to  50  ppm.   Blood  levels of  the 50  ppm group were not significantly
higher than  In the  10 ppm  dosed  birds.  No brain  residues were noted at the
10 ppm level.  Carcass  levels were  higher  for  the 50 ppm treatment compared
with 10 ppm.

    Mammals.   Lackey (1949a)  administered doses  of  4  g toxaphene  1n  corn
oil/day 1n  a  gelatin capsule  to  dogs  and  determined that brain tissues  (and
not fat) at day 106  after treatment showed Increased organic chloride levels
relative  to  unexposed animals.   This  was  Interpreted by  the  author  to  sig-
nify a storage of  toxaphene or  chlorinated derivatives  1n the brain but not
In the  fat.   In sheep,  steers and  dairy  cows, storage  of  toxaphene 1n fat
did  occur  on  repeated  exposure  (D1ephu1s  and  Dunn,  1949;  Conley,  1952).

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I—«
I—*
*—•I
                                                      1ABLE  III-2

                 Distribution of Toxaphene Continuously Administered Orally to Black Ducks from Birth
                   by Nixing  Toxaphene  In  Propylene  Glycol,  which Constituted IX of  the  Duck  Nash*
Dose (ppra
toxaphene In
the diet)
10





50





Nean Concentration (rag toxaphene/kg
Age
Week 2

Week 12

Nonth 19

Week 2

Week 12

Nonth 19

Gender
N/F

N/F

M (6)
F (6)
N/F

N/F

H (6)
F (6)
Generation
Fla
Fib
Fla
Fib
FO
FO
Fla
Fib
Fla
Fib
FO
FO
Carcass
6.3*0.3
3.1*1.3
7.7*0.9
3.U0.2
3.1*0.6
3.0±0.04
20.8*3.0
7. OM. 4
28.0*3.5
11.6i3.4
28.0*5.7
11.U1.4
Liver
NH
NN
NN
NN
0.40*0.08
0.40.*0.07
NN
NN
NN
NN
6.1*1.3
2. 2^0. 4
Brain
NN
NN
NN
NN
<0.1
<0.1
NN
NN
NN
NN
1.4*0.6(5)
0.68±0.45
ww) In

Blood
NN
NN
NN
NN
0.14*0
0.18M)
NN
NN
NN
NN
0.50*0
0.29^0

I


.01
.02




.16
.03
     •Source:  Haseltlne et al.. 1980

     NN = Not  measured
CO

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Feeding  toxaphene at  10 ppm  In  the diet  of  sheep  and  calves  for  28  days
revealed  no  detectable storage In  the  fat (Claborn,  1956).   Feeding cattle
25 and  100 ppm  1n the diet  for 16 weeks resulted In the appearance of. 12 and
38  mg  toxaphene/kg  omental  fat,  respectively.   Eight  weeks after  feeding
stopped,  storage  In  the 100  ppm  diet group  diminished  to  0.5  mg/kg  ww.
Dermal  spraying  (0.5X solution)  led to the appearance of toxaphene residues
In cattle  fat  of 2.5 mg/kg ww after a  single  spray (Claborn,  1956).   Analy-
sis of  fat  4 weeks after 1-2  sprayings of  0.5X solution  at 2-week Intervals
showed  toxaphene present 1n renal  fat.   Six  weeks  after  the  last spraying,
no  toxaphene could  be detected  In  fat.    Roberts  and Radeleff  (1960)  sug-
gested  that  meat from toxaphene-treated swine  1s  safe for  human consumption
If the  animals  are slaughtered at least 6  weeks  after being sprayed  once or
twice with a 0.05X solution of toxaphene.

    Clapp et al.  (1971)  reported  the results  of a 12-week feeding study with
12 male and  12  female rats  provided with  feed  containing 0,  2.33, 7, 21, 63
and 189  ppm toxaphene.   TLC  was used  to  assess  levels  1n omental  fat  and
liver.   Levels  In  .the  liver were  greatest  at   week  4,  but  fat  levels
increased continuously, although male rats  tended to have maximum fat levels
at week 8.   Below the 21 ppm  dose,  residues  1n  fat,  liver  and  carcass  were
similar.  Above  this dose, fat residues were proportional  to dose.

    Crowder  and  Dlndal  (1974)  studied  the distribution  of 20  mg technical
36C1-toxaphene/kg  bw administered  orally   by  stomach  Intubation  1n  0.5 ma.
of a peanut  oil/gum  acacia  solution  to  30-day-old Holtzman albino rats.   The
uptake of the label  In  various  tissues  from 3 hours to 20 days  (Table III-3)
showed that  the  highest  concentrations  In  most  tissues occurred  1n 12 hours.


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o
ro
                                                                             TABLE 111-3


                                         Percent  Uptake of Radioactivity In Various Rat Tissues and Organs following a  Single

                                          Dose of »*Cl-Toxaphene (20 mg/kg) Administered In O.S ML Peanut Oil/Green Acacia*
Tissue
Blood Supernatant
Blood Cells
StoMch
Liver
Kidney
fat
Testes
Brain
Muscle
Saall Intestine
First 2 CM
Last 2 CM
Large Intestine
Esophagus
Splten
X Tissue Total
Feces
Urine
X Excretion Total
Total X Recovered

0.125
0.64
3.1
3.7
0.33
O.OS
0.14
0.02
0.03
0.93

0.06
0.10
0.19
0.04
0.04
9.4



9.4

0.25
1.2
0
19
1.1
0.13
0.15
0.08
0.06
1.6

0.34
0.34
0.60
0
0.06
24.3



24

0.50
2.4
0
77
2.3
0.43
0.86
0.28
0.23
5.3

0.34
0.28
1.2
0.04
0.08
90.9



91

1
1.3
0.06
2.0
0.50
0.10
0.57
0.06
0.05
1.3

0.05
0.13
0.19
0.03
0.05
6.4
24
1.5
25
31

2
0.60
0.90
0.63
0.31
0.03
0.31
0.04
0.04
0

0.01
0.01
0.08
0.01
0.02
3.0
7.5
3.2
11
14
Days
3
0.36
2.6
0.61
0
0.03
0.18
0.03
0
0.65

0
0.15
0.02
0.01
0
4.6
1.3
2.9
4.1
8.7
Post-Dosing

456
0
0
0.
0.
0.
0.
0.
0
2.

0
0
0.
0.
0.
3.
1.1 1.
2.4 1.
3.5 2.
6.
.
-
39
01
01
18
03
-
4

-
-
03
02
03
1
1 1.2
8 1.2
9 2.4
0

7
0.18
0
0.16
0
0
0.02
0.02
0
0.40

0
0
0.04
0
0.24
1.1
0.69
1.2
1.8
2.9

8 9
0.09
1.1
0.12
0.48
0.03
3.7
0.06
0.01
0.14

0.84
0
0
0.03
0 0.06
6.6
0.27 0.31
0.54 0.72
0.8 1.0
7.6

20
0.06
1.2
0
0
0 ,
0.03
0
0
o.ai

0.09
0
0
0
0
2.2




           •Source:   Crowder and Dlndal. 1974.
o
*•
\
CD

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All  blood levels  of  labeled  toxaphene  peaked after  3  days.   The  data  may
Indicate  the distribution  of  toxaphene metabolites  1f  metabolism  Is  fast,
rather than  the  distribution of  toxaphene  Itself.   In addition,  If extensive
dechlorlnatlon occurred  the  labeled  Cl  detected  might represent  Inorganic Cl
rather than  organic CT.  The  former appeared to  be  the case for  the  feces
and  urine.   It  appears  that the 36C1  1s  rapidly secreted with  the stomach
acids within 0.50  days  after dosing,  which  Implied that  extensive metabolism
had occurred.

    Male  Sprague-Dawley   rats  (250-290 g)  were  administered   the  3«C1-
labeled compounds  by stomach Intubation  at  0  and -13 mg  toxaphene/kg bw with
0.15  ml  corn  oil  as  the  vehicle  and   with  another  0.10  ml  as  rinse
(Ohsawa  et   al.,  1975).   14C-toxaphene  was  similarly   administered at  0,
8.5,  12.7 and  19.0 mg/kg.   In the  same study 14C-labeled Toxicants  A  and B
were  administered  to  200-225  g  rats  at  0 or 0.84  and   0  or  2.6  mg/kg  bw,
respectively,  using 0.25  mi  corn  oil  as  the  vehicle   and  also  as  rinse.
The  results  given  In Table  III-4  show that toxaphene and Toxicants  A  and B
were  rapidly eliminated  (high excretion percent)  as metabolites,  and  that
toxaphene did  not  remain  as such In  tissues  to any  great  extent (14C- and
36C1-toxaphene levels  were  not  comparable).   Most of  the label  resided In
blood, fat,  liver   and  kidneys.  The  36Cl-results are  generally consistent
with  those  of  Crowder  and  Olndal (1974)  although detectable levels  In  the
blood and muscle were  not observed.   At  day  14  between   2.2 and  6.6% of  the
3«Cl-label  should  be  found  In  the  tissues  (Crowder  and Olndal,  1974)  and
>53X  should  have  been  excreted.  Since 59X  of  the  19  mg  14C-toxaphene/kg
dose  of  toxaphene  was  excreted to day  14  1n  the Ohsawa  et al.  (1975) study,
results  of  both studies  are  compatible.   Toxicant  A and  B appeared  to be


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o
I\J
o

o
                                                 TABLE  1114


     Distribution  of  Radioactivity  in  Various  Rat  Tissues  and Organs  at  Day  14 Following a Single Dose

     of a'Cl-Toxaphene, 14C-Toxaphene,  Toxicant A and B Administered In Corn Oil  by Stomach Intubation3
Tissue
Blood
Liver
Kidney
Fat
Bone
Brain
Heart
Lung
Nuscle
Spleen
Testes
Urine0
Feces0***
Breath »«C02C
X Excreted0

19 rag/kg
NR
0.30
NR
0.78
NR
NR
NR
NR
NR
NR
NR
31.8
27.1
NR
58.9
14C-Toxaphene
12.7 mg/kg
NR
NR
NR
NR
NR
NR
NR
NR
NR
NR
NR
25.4
31.7
NR
57.1

8.5 mg/kg
0.14
0.12
0.17
0.52
0.02-0.09
0.02-0.09
0.02-0.09
0.02-0.09
0.02-0.09
0.02-0.09
0.02-0.09
21.3
34.7
1.2
57.2
a*Cl-loxaphene
14.2 mg/kg
NR
NR
NR
NR
NR
NR
NR
NR
NR
NR
NR
49.1
26.9
NR
76.0
Levels (mg
Toxicant A
NR
NR
NR
NR
NR
NR
NR
NR
NR
NR
NR
28.3
47.8
1.8
68.5
label/kq uw)
Toxicant Bb
0.07
0.12 '
0.09
NR
<0.03
<0.03
<0.03
<0.03
<0.03
<0.03
<0.03
26.7
47.8
0.7
75.2
in
\
CD
aSource: Ohsawa et al.. 1975


bAt day 9


cPercent cumulative


dMethanol soluble fraction only


NR = Not reported

-------
quickly  dechlorlnated.    l4C-Tox1cant   8   was   hardly   detectable  In  most
tissues at day 9 except for blood, liver and kidney {Ohsawa et al., 1975).

    These results are  somewhat  different  from  those  described 1n the earlier
report of Clapp et  al.  (1971)  In  that  no deposition  1n fat was observed, but
the  TLC  technique  utilized  by  the  latter  authors may  not  have  been  as
specific as  the  techniques utilized  by Crowder  and  Dlndal  (1974)  and  Ohsawa
et al. (1975).

    Hale albino  Sprague-Dawley rats  were  separately administered toxaphene
(13  mg/kg   bw);   14C-toxaphene  (1.5  mg/kg  bw)  and  3.1  mg/kg bw  each  of
Toxicant B;  2-exo,5-endo,6-exo,8,9,10-hexachlorobornane  (CAS RN 57981-29-0);
and   2-endo,5-endo,6-exo,8,9,10-hexachlorobornane  (CAS  RN   65620-64-6);   a
mixture of the latter  two  compounds  (0.52/0.95  mg/kg bw, respectively) using
soybean oil  (0.15  mi)  as  a  vehicle for  administration by  stomach  Intuba-
tion;  and  0.1  mi  for  the rinse  (Saleh  and   Caslda,  1978).   GC  analysis
confirmed  that  all  of  these  compounds  underwent  dechlorlnatlon, and  that
chromatograms derived  from  samples  of  fat did  not  resemble  the toxaphene
standard.   This  was not the  case for  the  liver and feces.  The  amounts  of
Toxicant 8 and Us  metabolites  1n fat  and liver at 7 and 72 hours  after oral
Intubation are provided In Table  III-5.

    Peakall  (1979)  administered an acute single  dose of 120 mg toxaphene/kg
bw orally  1n a gelatin  capsule to male rats.   Four  animals were  sacrificed
at 1,  5  and  15  days.  The liver residues were  2.3,  4.2  and  5.7 mg/kg ww,
respectively.  The  residues 1n  brain were  below  detection  limits, 2 and 2.7
mg/kg ww,  respectively.   When  fed 2.4  mg  toxaphene/kg  bw/day,  four  animals


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o
o
o
                                                           TABLE  II1-5

                            Distribution and Metabolism of Toxicant B (51775-36-1) at 7 and 72 Hours
                                After Stomach Intubation of Sprague-Oawley Rats at 3.1 rog/kg bw*
uQ/kg ww of CAS RN
Tissue
Fat

Liver

Tine (hours)
post dosing
7
72
7
72
Number
of Rats
3
5
3
5
51775-36-1
453*285
335±44
8.6*4.1
17. 3H1. 7
65620-64-6
8±?
13*5
5.9H.9
2.0^0.6
57981-29-0
14il3
34±8
10.2i2.3
9.2i7.0
1
64618-63-9
15*9
14j;2
3.4±0.9
0.8±0.6
          'Source: Saleh and Caslda. 1978
o

00

-------
were  sacrificed  at  1,  3  and 6 months.   The "Mver residues were 28, 26 and 26
rag/kg  ww,  respectively.   Brain  residues  were  9.3,  13  and  12  mg/kg  ww,
respectively.  These  results  are  suggestive  that steady  state plateay levels
can be reached 1n liver and brain 1n chronic studies.

    In another study,  virgin  female  Sprague-Oawley  rats  (318+34 g) were used
and  pregnant animals  were  acutely  administered 2.6 mg  14C-toxaphene/kg bw
1n  olive  oil  (0.1  ml)  as  a  vehicle  for  oral gavage  (Pollock  and  Hill-
strand, 1982).   Rat chow and  water were available ad libitum.  The amount of
label  1n  various  organs,  excreta,  fetus  and  umbilical  cord  was  measured.
The results  are  provided 1n Table III-6.   The urinary excret.lon of the label
by  pregnant animals  up to day  5   compared  well with  that  of  nonpregnant
females  (Pollock and  Hlllstrand, 1982).   The  fecal  excretion (28.3%)  was
slightly  lower  than that found  1n  nonpregnant  animals  (35.7%).   The levels
In  the  fat  for  pregnant animals  at  day 5 (7.1-7.8  mg toxaphene/kg  ww)  also
agreed well  for  levels  1n  nonpregnant  animals after 7  days  (6.4  mg  toxa-
phene/kg  ww).   All  other tissues contained <10% of the level  In  fat.   The
carcass,  cecum,  small  Intestine  and adrenal  gland   contained  levels  >0.5 mg
toxaphene/kg ww  at  day  5.  There were Indications  that  metabolites resided
In  fat  deposits  and  1n  the  fetus,  but these were  not  specifically Identi-
fied.  Transplacental  transfer  (<1%  of  the administered  dose) also occurred,
as  did  blood-brain  barrier  transfer.   From  Table III-6, the adrenal gland,
carcass,  cecum,  abdominal   fat,  GI  tract contents,  kidney,  large  Intestine,
liver, ovaries and  small Intestines  each  contained  over  1 mg toxaphene/kg ww
at  day 1.   At  day 3,  only  the  adrenal  gland,  carcass,  cecum, abdominal fat,
GI  tract  contents  and  small  Intestines  contained  residues  >0.5  mg  toxa-
phene/kg ww.


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                                 TABLE III-6
        Distribution of_Rad1oact1v1ty 1n Various Female Sprague-Dawley
           Rat Tissues and Organs Following a Single Dose of 2.6 mg
                14C-Toxaphene/kg bw Dose to Four Pregnant Rats
              Administered 1n OUve 011 (0.1 mi) by Oral Gavaged
                                    14C-labeled Material (pg Toxaphene
                                       Equivalent/kg Wet Weight)  at
Tissue
Adrenal gland
Brain
Carcassb
Cecum
Fat (abdominal)
Fetus
GI tract0
Heart
Kidney
Large Intestine
Liver
Lungs
Ovaries
Placenta
Small Intestine
Spleen
Stomach
Umbilical
Uterus
Ur1ned
Fecesd'e
Day 1
1092
185
1532
6354
7208
84
6534
349
1195
2757
1210
735
1054
191
6890
171
709
207
154
(8.9+2.0)
(9.3+4.8)
Day 3
577
63
588
1225
8474
30
1995
90
317
370
397
413
k 175
70
629
49
327
145
99
18.3
23.8
Day 5
625-731
57-62
489-924
1012-1136
7107-7845
27-29
NM
71-92
303-347
204-428
363-384
339-407
469-484
47-56
486-1185
50-73
180-351
134-155
56-216
22.0
28.3
aSource: Pollock and Hlllstrand, 1982
^Contains blood and subcutaneous fat
cContents of GI tract at termination
^Percent cumulative for four animals per day
emethanol soluble radioactivity
NM = Not measured
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Distribution  In Human Tissues
    Apart  from the  Instances  noted  In  the  absorption  section,  no  residue
data,  or metabolites  of  toxaphene  have  been  found  1n  human  tissue,  even
though  44  cases  (Including deaths)  Involving exposure of humans  were  docu-
mented  between  1966 and 1975  In  the United States  (U.S. EPA,  1977).   In an
occupational  study,  exposure of  54  workers  primarily  by the  Inhalation or
dermal  routes  did  not generate detectable levels of  toxaphene  1n the  blood
(>0.1 mg/kg  blood)  1n spite of the  fact that 49  of  the  personal air  samples
showed detectable amounts of toxaphene (U.S. EPA, 1980).

    The  above  findings  signify  that  toxaphene may either be  quickly  metabo-
lized and  transported In another  form In  humans, or  that no  absorption  has
occurred.  In  view  of the  many human  and  animal  poisoning  Incidents  Involv-
ing  toxaphene,  H  1s more  likely  that toxaphene  1s  metabolized quickly  and
then transported.

Metabolism
    Toxaphene  undergoes  fast  reductive  dechlorlnatlon,  dehydrochlorlnatlon
and  hydroxylatlon   1n mammals.   Little   Is  known  about  the  mechanism  of
transport.

    Toxaphene was thought  to  be slowly detoxified In  such  animals as sheep,
steers and cows  by  excretion of ethereal  sulfate and "glucuronate" (Conley,
1952).   Crowder   and   Dlndal   (1974)  In   their  distribution   study   of
"Cl-toxaphene reported  1n the section  on "Distribution In  Animal Tissues"
observed  much  redistribution  of   »*C1   label   with  time.    Host  of  the
3*Cl-labeled  toxaphene  was excreted  (see  Table  III-3)  within  6-7 days  and
little remained In the tissues.

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    Ohsawa  et  al.  (1975)  confirmed the observations  of  Crowder  and  Dlndal
 (1974)  with 3«Cl-toxaphet)e  (see  Table  III-4).   The only  Identified  metabo-
 lite  was  chloride  Ion  that  appeared  almost  entirely   In  the  urine  and
 accounted  for  about half  of 3*Cl-toxaphene eliminated.   The  dechlorlnatlon
 occurred  mostly  within two  days.    14C-toxaphene  distribution  as  well  as
 that  of  Toxicants A and  B showed  only  a  small  amount unmetabollzed  In  the
 feces  (3X  for  14C-toxaphene)  and   that  the  metabolites  probably  Included
 acidic  materials  partially  or  completely  dechloMnated,  and  14CO?   (2%  of
 14C-toxaphene).   The ease  of dechlorlnatlon  appears   to  be  shared  by  all
 components of toxaphene, even Toxicants A and B.

    Khalifa et  al.  (1976)  examined  the reactions of toxaphene  and Toxicants
 A  and B  with   two  Iron  (II)  protoporphyrln  systems,  hematln  reduced  with
 sodium thlosulfate and  mlcrosomal cytochrome P-450  reduced with NADPH.   They
 found that  toxaphene reacts with reduced  hematln 1n neutral  aqueous  medium
 to cleave  about  half of  the C-C1  bonds,  yielding derivatives  with  shorter
retention times on  gas  chromatography and of reduced  sensitivity  for  detec-
 tion  by  electron capture.   The  system  also  converts   Toxicants  A and B  to
products  formed  by  reductive  dechlorlnatlon,   dehydrochlorlnatlon   and  a
combination  of  these   reactions.    Extensive  metabolism  of  toxaphene  and
Toxicants A and B by rat liver mlcrosomes required NADPH.

    From these  results  It  Is  clear  that  many  toxaphene components, Including
Toxicants A and B, are  converted  by one  or  both  of these Iron (II) protopor-
phyMns   to  more polar   derivatives  (as  evidenced by TLC).  The  studies  with
02010                               111-20                           02/25/87

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 reduced  hematlh  establish that  toxaphene  undergoes  extensive dechlorInatlon
 and  that the  products  from  Toxicants  A  and  B  determined  by  GC-CI-MS  are
 formed by  two  or more of the  following:   reductive  dechlor1nat1on,  dehydro-
 chlorlnatlon and vicinal chloride elimination.

    Toxaphene  has  been shown  to yield  type  I binding  spectra  with  hepatic
 cytochrome  P-450  of  rats,  mice,  sheep  and  rabbits,  which suggests  that
 toxaphene  may  serve  as  a  substrate  for  the  hepatic mlcrosomal MFO system
 (Kulkarnl et al., 1975).  Type II binding has not been observed.

    In another  rat metabolism study,  Chandurkar  and  Matsumura  (1979a)  found
 that  dechloMnatlon  and  oxldatlve  degradation  by  MFO  Involving  cytochrome
 P-450 are active degradation mechanisms for toxaphene.

    Toxaphene metabolism  1n  chickens  and  6  species  of  mammals  (guinea  pig,
 hamster,  rabbit, mouse, rat,  monkey)  Involves  extensive dechloMnatlon;  and,
 1n the case of rats,  ~50% of  the dose 1s excreted as chloride 1on (Saleh and
 Caslda, 1979; Saleh et al., 1977, 1979).

    Saleh  and  Caslda  (1978)  showed  that  the major  products of Toxicant  B
 produced by  rat  liver mlcrosomes are CAS RN  57981-29-0 and  65620-64-6  In  a
 2:1.  Table  III-7  lists  synonyms, chemical names and  Indexing  terms  for the
major  metabolites  and  degradation  products of  Toxicant B.   These  products
were  not detected  under  aerobic conditions  or  under aerobic  or  anaerobic
 conditions  1f  NADPH was absent.  These products were  also  found  (see Table
 III-5) 1n  fat,  liver and feces  1n  addition to CAS  RN 64618-63-9 after  oral
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                                                                             1ABIE  111-7

                                                           Identified Metabolites of loxaphene Components
o
to
o
           CAS RH
             NT
Chemical Abstracts Indexing lerm (9C1)
                Synonym
                                                                                                 •nls
                                                                                                                                Reference
          $7981 WO
o

o
*.
\
GO
          64618-63-9
          CIOMIOC'6
          6S620-64-6
          66157-70-8

          C10H11C15
           70459-31 3
Blcyclo[2.2.1)heptane.2.3.6 trtchloro
l.7.7.-trls(chloromethyl) ,(2-exo.3 endo.
6-exo)-
                           or
                           2-exo.S-endo.6-exo.8.9.10-hexachloro-
                           bornane
Blcyclo(2.2.1]hept-2-ene,2.5.6-trIchloro-
1.7.7-trts(chloromethyl)-.5 (endo.6 exo)-
                           or
                           2.5-endo.6-exo.8.9.10-hexachloroborn-
                           2.3-cne
Blcyclo(2.?.l)heptane.?.3.6. trlchloro 1.
7.7 trts(chloromethyl) ,(2-exo.3 endo.
6-endo)-
                           or

                           2-endo.S-endo.6-6X0.8.9.10-hexachloro-
                           bornane
Trlcyclol2.2.1.02.*]heptane.3.4-dlchloro-
1.7.7-trls(chloromethyl)-.

or
2.5-endo.8.9,10-pentachloroiricyclene

Blcyclo)2.2.1Jheptane.2.3.5.5 tetrachloro
7.7-bts(chloromethyl)-l-(d1chloromethyl)-
(exo.exo)-
or

3.3.5.6-exo-8.9.10.10-octachlorobornane
                                                                             Reductive dechlor(nation of toxaphene
                                                                             toxicant B In: reduced heaatln system.
                                                                             bovine rumen flutd. rat liver micro-
                                                                             somes . and rats Jn vivo
                                                                             fecal metabolite of toxaphene toxicant
                                                                             B In: chicken, mouse, rat. hamster.
                                                                             guinea pig. rabbit and monkey

                                                                             Reductive dehydrochlorInatton of
                                                                             toxaphene toxicant B In: reduced
                                                                             hemattn system and rats \n vivo
                                                                             Fecal metabolite of toxaphene toxicant
                                                                             B In: chicken, mouse, rat. hamster.
                                                                             guinea pig. rabbit and monkey

                                                                             Reductive dechiorInatton of toxaphene
                                                                             toxicant B In: reduced hemattn system.
                                                                             bovine rumen fluid, rat liver micro-
                                                                             somes, and rats In vivo
                                                                             Fecal metabolite of toxaphene toxicant
                                                                             B In: chicken, mouse, rat. hamster.
                                                                             guinea pig. rabbit and monkey

                                                                             Reduced hemattn conversion of
                                                                             toxaphene toxicant B
                                                                             loxaphene toxicant C metabolite In rat
                                                                             liver In vitro
Saleh and Caslda. 1978
                                                                                               Saleh et  al..  1979
Saleh and Caslda. 1978
                                                                                               Saleh  et  al.  1979
Saleh and Caslda. 1978
                                                                                               Saleh  et  al..  1979
Saleh and Caslda. 1978
Chandurkar and Hatsumura.  1979b
          CAS RN = Chemical Abstracts Service Registry Number; Nf -- Molecular lornula; 9CI -- Ninth Collective Index.  Chemical  Abstracts

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administration  by  gavage of 3.1 mg Toxicant  B/kg  bw to male albino Sprague-
Oawley strain rats.   The-rat1o  of  CAS RN 57981-29-0 to 65620-64-6 was 1.8 In
fat,  and  1.7 1n  the  liver 7 hours after  administration'.   At  72  hours,  the
ratios were  2.6 and 4.6,  respectively, and  2.5  1n  the feces.   At  least 7.4%
of  the administered dose was eliminated after 72  hours 1n rats by reductive
dechlorlnatlon  at  the  germinal dlchloro  group.   The chromatograms  of  the
residues  In  fat  almost  conformed to  the toxaphene  standard but chromatograms
of  liver  and feces samples  did not.   The latter samples,  however,  did con-
tain toxaphene components.

    Chandurkar and  Matsumura  (1979a)  reported that  In  the  presence of NAOPH,
the   liver   mlcrosomal   fraction   of   rats  metabolized  3*Cl-toxaphene  and
14C-toxaphene after 2 hours  at  37°C  to the  extent of 22  and  9.5%,  respec-
tively.   In  the  presence of glutatMone  (GSH),  the  extent  of  metabolized
3*C1  and  14C toxaphene were 11  and  4.8%,  respectively;   and  with  cystelne
as  a  cofactor,  the respective values were 10 and  3.6%.  Inhibitors  of NADPH
dependent   mixed-function   oxldases   and   of   glutath1one-S-transferases
Inhibited conversion  of  toxaphene  to polar  water-soluble  metabolites.   The
results depended  on which fraction was utilized from  rat  liver homogenates.
For  example, one  NAOPH  dependent  mixed-function  oxldase  Inhibitor  caused
100%  Inhibition  of conversion  by  a  lOO.OOOxg supernatant  fraction  compared
with  53%  utilizing a  20,000xg  supernatant  fraction from  the  same  homogen-
ate.   When  Toxicants   B  (Gig)   and  C   (Clg)   were  Investigated   In  the
20,000xg  supernatant  under the  same  conditions as  toxaphene,  the  extents of
metabolism were ~73  and  56%,  respectively  In  the  presence  of  NAOPH;  the
respective figures  for  glutathlone were  31  and 47%.   Whereas  the extent of
Toxicant  C metabolism In  the presence of NAOPH was complete  within 1 hour,


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 the  rate of Toxicant  8  conversion was constant  with  time to 3  hours.   The
 control  showed  an  ability  to  metabolize  Toxicant B  and  C without  added
 NADPH, Toxicant  C  being  more prone 1n  this  regard.  TLC  then EC/GC revealed
 that  different  components  In toxaphene were metabolized by  the  NADPH system
 as compared  with the  GSH system.  Most of  these  metabolites  were more polar
 than  the parent compounds  {TLC  data),  some  being hydroxyllc  (esterlf1cat1on
 data), and  some being  still chlorinated  (EC/GC  data).   Treatment  of  water
 soluble  14C-toxaphene  metabolites with B-glucuron1dase  1n  the  presence  of
 NADPH revealed  an  Increase of  16.IX  1n ether extractable  material originally
 present  as  glucuronldes.  Smaller  Increases of  7.8 and  8.6X were  shown  In
 the  presence  of  B-sulfatase  and  dilute  hydrochloric  add,  respectively.
 Addition  of  UDP-g1ucuron1c  add to  a  toxaphene/NADPH  system  Increased  the
 extent of  metabolism  8-fold, but  not 1n  the additional presence  of  a NADPH
 dependent mixed-function oxldase Inhibitor.  These results  show  that ox 1 da-
 tive  metabolism Is Important  for  toxaphene.   The  water-soluble  metabolites
 of GSH-fort1f1ed Incubates are  probably GSH-conjugates  formed Involving loss
 of a chlorine from the original  toxaphene components.

    In another  report  of  Chandurkar  and Matsumura  (1979b),  using  the  same
 20,000xg  supernatant  system   fortified   with   NADPH   (see   Chandurkar  and
Matsumura, 1979a), the major dechlorlnated  metabolite of  Toxicant C In vitro
was  shown  to  have  a  molecular   formula   of  c-|QHincl8  ^MS  data).   This
 compound has been  designated as 3,3,5,6-exo-8,9,10,10-octachlorobornane (CAS
 RN 70459-31-3)  (see Table III-7).  Oxidation  products  of Toxicants  B and C
were  not tertiary  alcohols,  but  4  products  of  Toxicant C  were secondary
alcohols,  and  one  was  a  primary  alcohol.  No Toxicant   B  metabolites  were
 primary  alcohols.   None  of the  secondary alcohols  were dlols.   NADPH caused


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 more Toxicant B  metabolism than for Toxicant  C,  but no alcohol metabolites
 of  Toxicant  B could  be-found, thus being  suggestive of  the occurrence  of
 reductive dechloMnatlon rather than hydroxylatlve dechlorlnatlon.  This may
 Indicate different  loci  for metabolism  of  Toxicants B  and C.

     Pollock  and  Hlllstrand  (1982)  reported  that the  metabolites measured  In
 the  fetus  are   apparently  different  from   those  measured   In  the  fat   of
,Sprague-Dawley dams based on TLC analysis.  No  metabolite  Identification was
 reported.

 Elimination
     In   a  study  where White  Leghorn  chickens  were   fed  5,  50 or  100 ppm
 toxaphene 1n  the diet for  34  weeks (Bush  et   a!..  1977),  the half-life  of
 toxaphene 1n  adipose  tissue was 42,  25  and 20 days, respectively.

     To  determine the  length of time necessary for  depletion of  36Cl-toxa-
 phene residues from  adipose tissue of broiler  chickens,  15 birds  from each
 of  the  replicates  (45  birds/treatment group)   were  fed  toxaphene-frce feed
 from week 6  onwards  (Bush et  al.,  1978).   The  half-life  of  toxaphene  In
 adipose  tissue was found to be 2.66, 2.76,  2.47 and 2.5 weeks  for  birds fed
 0.22,  0.40,   2.16 and  3.82 ppm toxaphene In  the  diet,  respectively.  Data
 were obtained from three birds, each sacrificed at Intervals of 2  weeks for
 10 weeks.

     Black ducklings were  fed  dietary  levels of 0, 10 and 50  ppm  technical
 grade toxaphene  In  propylene  glycol  (1%  of a  commercial  duck feeder  mash)
 until  the egg laying  season  (1976) was  over   (Haseltlne  et  al.,   1980).   A


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select  number  of  eggs  (Fg  generation)  were allowed  to hatch,  and the  F,
generation ducklings were_s1m1larly exposed  up  to  the  next  egg laying season
(1977).   In  1976,  the  six eggs  from  the FQ generation ducks  samples  con-
tained  4.4^0.4  and 23.5^1.9 mg/kg ww  for the 10 and  50 ppm concentrations,
using total area  GC estimation.   In 1977,  the respective figures for the two
levels were  Increased  to  5:6^0.6 and  29.5+2.0 mg/kg ww.  In 1977  the levels
In both  the  eggs  and  adult carcases  were similar  except  1n  females fed  50
ppm  toxaphene.   Carcass  levels for females  were 11.U1.4 compared  with  egg
levels of 29.5^2.0.  The  Investigators suggest  that  residues are not concen-
trated  In eggs,  since  eggs contained  a similar  toxaphene load as  carcasses.
Both adults and  young birds  can  metabolize toxaphene and excrete enough  that
little accumulation takes  place In the organs.

    The  half-life of  14C- or  3*Cl-labeled  toxaphene   In  rats  after  single
oral   doses  appears to  be  less  than  a  week,  with  most of  the elimination
occurring 1n  the urine and  feces  (Crowder  and  Olndal,  1974;  Ohsawa  et  a "I.,
1975).  Only  a  small  portion  of  the  urine and  fecal  metabolites  are elimi-
nated as  glucuronld-e  or  sulfate  conjugates  (Chandurkar, 1977),  In  contrast
to early findings by Conley (1952).

    Ohsawa et  al.  (1975)  found  that 49%  of  »*Cl-label was  eliminated  In
the  urine 14  days after  administration  of  14.2 mg  3«Cl-toxaphene/kg  bw
(see  Table  III-4);  In the same  time  25.4% of  the  label  from  a  12.7  mg
14C-toxaphene/kg  bw dose was  eliminated 1n  the  urine.    The  bulk of  the
"Cl-label (90%) was   chloride 1on.   After  14   days,  27% of  "Cl-label  was
found 1n  the  feces.  Thus,  1n 14 days,   76% of 3*Cl-label   was  found 1n the
urine  and  feces.   Subfractlonated   toxaphene   gave  essentially  the   same


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 results.   Unmetabollzed  toxaphene comprised 3.4% of  the  feces  residues,  and
 around  27% 1n  urine and-feces appear  to  be organic*  that are dechloMnatlon
 products.  Between  5 and 10% are completely dechlorlnated.

    The  data for  Toxicants  A  and  8  are compared with  that  for  14C-toxa-
 phene  In  Tables  III-4 and III-8.  In  general,  Toxicants  A  and  B appear more
 readily  eliminated  but their dosages  were  lower than  for  toxaphene  Itself,
 and only  one animal was  tested.   Unmetabollzed  compound constituted only  8.1
 (Toxicant  A) and 2.6%  (Toxicant B) of fecal residues,  respectively.

    In  a  similar  set of  experiments  (Saleh  and  Caslda,   1978),  a  1.5  mg
 14C-Tox1cant B  (CAS No.  51775-36-1)/kg bw  dose was  given  to Sprague-Dawley
 rats  by  stomach  tube with  soybean  oil  as  the  vehicle.   Only  0.2^0.1%  of
 Unmetabollzed Toxicant B was  found  In the  feces  at  0-72  hours, along with
 2.U0.6,   5.3+1.6   and  1.0*0.3%  of  the  administered  label   as  CAS   RN
 65620-64-6,  57981-29-0 and 64618-63-9,  respectively.   The combination CAS RN
 65620-64-6  and  57981-29-0 constituted 7.4% of  the  administered label,  and
 constituted  products   of  reductive  dechlor1nat1on  at the  germinal  dlchloro
 group.  When a mixture of  CAS  RN 65620-64-6 and 57981-29-0 (0.95/0.52 mg/kg)
was administered  to  the  same  strain of  rat,  the  feces  contained  45-47% of
 the Unmetabollzed  compounds  collected up  to  72 hours.   The  feces  contained
many peaks of shorter  retention time than those 1n  toxaphene standards.

    Chandurkar and  Natsumura (1979a) also  utilized four  male Sprague-Dawley
 albino  rats  to  administer a dose  of 15 mg 14C-toxaphene/kg  bw  by  a stomach
 tube  (0.25 ml  of corn oil).   Fifty-seven percent  of  the label  was excreted
 In  the  feces.   Only 9% of the administered dose  was  found  1n the urine.   On


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                                  TABLE  III-8
     Elimination of 14C-Toxaphene, Toxicant A and Toxicant B Administered
       to Sprague-Qawley Rats  by  Stomach Intubation  1n  Corn  011  Carrier3
Label Eliminated
(Percent of Administered Dose)
Toxaohene
Sample
Urine
Feces
Breath
1*C02
Totals
Time After
Treatment
(days)
0-1
1-2
2-14
0-1
1-2
2-14
0-1
2-14

(8.5 mg/kg)
3 rats
10.6
6.3
4.4
34. 7b
0.8
0.4
57.2
(19 mg/kg)
2 rats
11.3
11.8
8.7
27. lb
NM
NM
58.9
Toxicant A
(0.84 mg/kg)
1 rat
18.9
6.0
3.4
27.4
6.0
5.0
0.8
1.0
68.5
Toxicant 8
(2.6 mg/kg)
1 rat
15.7
6.2
4.8
32.3
9.7
5.9
0.5
0.2
75.2
aSource: Ohsawa et al.t 1975
blt  Is  not  clear  1f  these  figures  are  totals  for  days  0-14  or  the  same
 measurement for each time after treatment.
NM = Not measured
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 the average,  8.9%  of  the  urine metabolites  and 0.7% of the fecal metabolites
 were  glucuronldes; 9.5%_jof  the  urinary  and  7.5X  of the  fecal  metabolites
 were  add-hydrolyable  products;  only a small  percentage  (0.7%)  of  the  total
 fecal metabolites  are sulfate conjugates.

    A  single  spray treatment  of cows with  2  quarts of  0.5%  toxaphene  sus-
 pended  1n  water did not  produce detectable toxaphene  In  the  milk  (Lelghton
 et  al.,  1952).  Spraying cows  twice  at  3-week  Intervals  with 2  quarts  of
 0.5%  toxaphene  emulsion or suspension  resulted  In  peak  levels  of  0.51-0.82
mg/i  milk  1  or 2  days  after  the  second  spraying.    Twenty-one days  after
 spraying had  ceased,  the toxaphene  concentrations  returned  to  preexposure
 levels  (Claborn,  1956).  Spraying  cows  under exaggerated  conditions  (twice
dally for  21  days  with 57 mg of  a  2X oil  solution  of toxaphene) resulted  In
peak  residues  of  0.11-0.50 mg/l  milk after -3  days.   Twenty-one  days  after
 spraying ceased,   the  concentration diminished  to  0.06 mg/i  milk  (Claborn,
1956).

    Keating  (1979) reported  on  toxaphene  residues  In  cow  milk   from  cows
dipped  once  or twice  weekly   1n  AU1k [0.25% w/w  toxaphene plus  0.03% w/v
D1oxath1on (Delnav)].   Residues  ranged  from 27-45 mg/kg  milk fat,  with  maxi-
mum residues  found one  day after dipping.   When  dipping was discontinued for
a cow dipped  In AH1k once weekly,  the levels decreased  to 5  mg/kg milk fat
after  19  days.  Even  at  32  days,  toxaphene  could  still  be  detected  (3.1
mg/kg).

    Pollock and Hlllstrand (1982)  found  that excretion  In urine  and  feces
 for pregnant  rats  (Sprague-Oawley)  1s very  similar  to  that In virgin female


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-------
rats,  although  there  was  a  large  weight  difference.   The  results  for
pregnant  females  are provided In Table  III-6, which  Indicate  a half-life of
-5 days.

    Toxaphene  was  administered   to  cows  continuously  at  Increasing  dally
concentrations  ranging from  1.7-12  ppm  diet.   Toxaphene levels  were  first
detected  1n  the milk  of  cows In the  4  ppm group after  3  weeks.   Excretion
equalled  7-8 mg  toxaphene/l  milk 1n  the 12 ppm  group.  When  exposure  was
discontinued,  the  concentration  1n  milk  diminished  promptly  (Shaw,  1947).
In  cows  fed  contaminated hay  for  16 weeks,  the  toxaphene  levels  )n  milk
(50-410 mg  toxaphene/l)  were  proportional to the  amount In  the  diet,  with
the  average   excretion  ranging  from 2.3-18.2 mg  toxaphene/l  milk  (Bateman
et al.,  1953).   Claborn  (1960)  fed  four  groups of  three cows  each  20,  60,
100  and  140  ppm  toxaphene   In  the  diet,  respectively.   The  excretion  was
proportional   to  the  toxaphene concentration.   Once exposure ceased, residues
diminished  to  13-17% within  1   week,  followed by  a  slow excretion  phase
probably related to the mobilization of fat reserves.

    Cows were  fed  0-20 ppm  toxaphene  1n the diet for  77 days; milk  samples
were  analyzed  twice  weekly  (Zwelg  et  al.,  1963).   The results  confirmed
previous  findings  at  higher  feed  levels  that  toxaphene  residues  declined
rapidly  after  toxaphene  exposure   had   terminated.    Residues  ranged  from
0.043-0.179   mg   toxaphene/l   milk,    and   were   concentration-dependent.
Following cessation  of exposure, residues 1n milk  decreased  to undetectable
levels after  2  weeks  for  levels  lower  than 10 ppm toxaphene.   For the 20 ppm
level, residues  were  detected  30 days  after  feeding  contaminated diet  was
halted.
02010                               111-30                           02/04/87

-------
    In  another  study  Involving  12  cows  fed  5  ppm  toxaphene  1n  the  diet
(Cairns  et  al., 1981),  toxaphene metabolites and  components were  found  In
milk  fat  with  the  EC/GC chromatograms  being  enriched In  the  components  of
shorter GC retention time.   Six  milk  samples  contained residues estimated  by
CI/SIM/GC/MS  to range  from between  7  and 280  mg/l  milk.   An EC/GC  study
using  toxaphene standards  estimated  the range  to  be 10-620 mg  toxaphene/l
milk.  The  noncorrespondence of  residues  to  Intake may  reflect the time  of
sampling  since  this  was  not  specified.   However,  this does  Illustrate  that
the  analytical  methods  for  toxaphene   may   yield   different  answers  when
enrichment or depletion of  selected compounds  1n toxaphene occurs.

    A  summary  of elimination  half-life  data   for  various animal  species  1s
provided  1n Table III-9.  Saleh  and  Caslda (1979)  reported that quick metab-
olism  and  elimination  occurs  1n  rats,  mice,  guinea  pigs,  hamsters, rabbits
and  chickens,  with  the  monkey  metabolizing  toxaphene most  extensively  and
rapidly,  and  chickens  the  least  and  slowest.   The  order  of  decreasing  rate
of  metabolism  1s  as  follows:   monkeys,  rats,  hamsters  >  mice,  rabbits,
guinea pigs > chickens  for  a dose of  Toxicant  B at 3 mg/kg bw.   The data for
toxaphene In Table  III-9 are In  general  agreement  with  the elimination  data
for Toxicant B, with half-lives being <1  week  for all mammals, Including  man.

Summary
 .   Though  no  precise   quantitative  data  are  available,  the symptoms  of
toxicosis as well as the presence of  residues  and  metabolites after exposure
1n animals, birds and humans Indicate  that toxaphene can  be absorbed through
the skin (especially 1f mixed with xylene), the lung, and the gut.
02010                               111-31                           02/04/87

-------
                                                TABLE  III-9



              Experimental or Calculated Half-Life of Toxaphene Elimination In Various  Species
Animal
Broiler chicken
(Hubbard-Hubbard)
Cattle
Cow
~ Rat (Holtzman)
i
Hale rat
(Sprague-Dawley)
Male rat
(Sprague-Dawley)
Male rat
(Sprague-Dawley)
Female rat
( Sprague-Dawley )
o Humans
^>
o
Dosea
(carrier)
0.22-3.82°
100° for 16 weeks
57 mg of 2X oil
solution sprayed
twice dally for
21 days
20 (peanut oil/
green acacia)
8.5-19
(corn oil)
0.52-0.95
(soybean oil)
15 mg
(corn oil)
2.6
(olive oil)
Catfish fillet
52 mg/kg fillet
Compound
»*C1 -label
Unlabeled
Unlabeled
•*Cl-label
"Cl-label
••Cl-label
Toxicants A.B
"Cl-label
Toxicant B
"Cl-label
"Cl-label
Unlabeled
Route Half-Life
(days)
adipose (18.2H)
adipose 5-6(calc.)
milk 21
feces; 6-7
urine
feces; <14
urine
feces 3
feces 4-5
feces; 5
urine
blood 6-7
Reference
Bush et al.. 1978
Claborn. 1956
Claborn. 1956
Crowder and Dlndal.
1974
Ohsawa et al.. 1975
Saleh and Caslda.
1978
Chandurkar and
Hatsumura. 1979a
Pollock and
Hlllstrand. 1982
U.S. EPA. 1978
CD
aln mg toxaphene/kg body unless specified otherwise



bppm diet

-------
    In  birds  (ring-necked  pheasants,  ducks  and  leghorn  layers),  toxaphene
can  be tolerated  for  months  with  the most  sensitive effect  being  altered
cartilaginous structures.   Extensive  dechlorlnatlon of toxaphene occurs with
chlorinated  organic  residues  occurring  mostly  In  the fat.   White  Leghorns
were able  to metabolize  or.  excrete  toxaphene  at a rate comparable with dally
Intake.  In ducks, brain residues appeared at doses >10 ppm In the feed.

    In mammals,  storage  of  toxaphene 1n the  fat  has  been reported In sheep,
steers  and dairy  cows.   Accumulation  In  cattle  occurred  at  exposures  to
25-100 ppm toxaphene In the  feed  for 16 weeks.   In  contrast,  dogs  appeared
to  store   toxaphene  derivatives preferentially   1n  the   brain.   In  male  or
female rats, fat  levels  of  toxaphene  were  relative to concentrations  >21 ppm
1n  the diet.   Studies with labeled toxaphene  revealed  quick dechlorlnatlon
and subsequent elimination  of toxaphene,  Toxicant  A  and  Toxicant  B  at doses
<19 mg toxaphene/kg  bw administered  by  gavage to male rats using corn oil as
the  carrier.   Toxaphene derivatives  resided  In  the  blood,  fat, liver  and
kidneys.   When  male rats  were  chronically dosed  with   2.4  mg  toxaphene/kg
bw/day, plateau  levels  were  found  In  the  liver  and  brain after 1,  3 and 6
months.   Transplacental  transfer  as well  as  blood-brain barrier  transfer
occurred  1n  rats.   The  adrenal gland,  carcass,  cecum and abdominal  fat  of
female rats  contained  toxaphene derivatives  3  days after  acute dosing with
2.6 mg toxaphene/kg bw.  Extensive  dechlorlnatlon and rapid elimination  of
metabolites  occurred  also   1n  guinea  pigs,   hamsters,   rabbits,  mice  and
monkeys.   No  residue data  or metabolites  of  toxaphene  In humans  have been
reported 1n the available literature.
02010                               111-33                           02/25/87

-------
    In  rats,  successful  detoxification appears to require  NADPH  and  oxygen.
Toxicants  A  and  B  underwent reductive  dechloMnatlon,  dehydrochloMnatlon,
and  vicinal  chloride  elimination.   Involvement of  the MFO  system  Is  sug-
gested  by  the observation of Type  I  binding spectra with  the  hepatic  cyto-
chrome  P-450  of  rats,  mice,  sheep and rabbits, as well  as  the  enhanced tox-
Idty  to  mice  of toxaphene  In  the presence  of  plperonyl butoxlde.   There
also appeared,  to be a detoxification  pathway  dependent on GSH.   Toxicant  B
resisted metabolism more  than  Toxicant C.  Glucuronlde (major)  and  sulfate
(minor) conjugates  have  been detected  as metabolites.  Toxicant  C  also pro-
duced  one  primary and  four  secondary  alcohols  whereas  reductive  dechlorlna-
tlon predominated for  Toxicant  B.   These data  suggest that toxaphene  com-
ponents are  not  metabolized necessarily at  the same loci  In  the mlcrosomal
system.

    Elimination  of  products  derived from toxaphene has  been  demonstrated  to
occur  In   the  feces,   urine  and  1n  expired air  of   rats.   The  fecal  route
appears to be  the more predominant route than  In  urine.   Milk  and eggs also
showed  residues.  The  species order of decreasing  elimination  efficiency for
Toxicant B Is  as follows:  monkey, rat, hamster > mouse,  rabbit, guinea pig
> chicken.
02010                               111-34                           02/25/87

-------
                              IV.   HUNAN  EXPOSURE







    This  chapter  will  be  submitted  by  the  Science  and  Technology  Branch,



Criteria and Standards Division, Office of Drinking Hater.
02020                                 IV-1                            02/25/87

-------
IV.  SOURCES OF HUMAN EXPOSURE - TOXAPHENE








     This chapter summarizes the available data on use,



environmental fate, and occurrence to characterize the



potential for exposure to toxaphene.  A more extensive



discussion of the information on toxaphene in the environment



is presented in the draft document entitled "Occurrence of



Pesticides in Drinking Water, Food and Air" (U.S.EPA, 1987).








     Humans may be exposed to toxaphene from a variety of



sources, including drinking water, food and ambient air. •



Individual exposure to toxaphene will vary widely based on



factors such as where a person lives, works and travels,



and what a person eats.  Intake of toxaphene will be affected



by age, weight-and lifestyle.








     The Exposure Estimates section of this chapter presents



available information on the range of human exposure to and



intake of toxaphene from drinking water, food and ambient



air.  It is not possible to provide an estimate of the number



of individuals experiencing specific exposures from these



sources.  However, this section provides some insight into



the sources' relative contributions.
                             IV-1

-------
A.  Use/Environmental Fate








     Toxaphene is a pesticide that was widely used during the



1960s and 1970s primarily on agricultural crops and livestock



In 1974, domestic production of the compound was estimated



at 103 million pounds, with approximately 74 million pounds



applied for agricultural uses (U.S. EPA, 1977).  By 1982,



domestic usage had declined to about 16 million pounds, with



only 5.9 million pounds used on field crops (USDA, 1983).



In November of the same year, EPA published in the Federal



Register its intent to cancel or restrict registrations for



products containing toxaphene.  All major uses of toxaphene



were cancelled (47 FR 53784)(USEPA 1982).   In addition, all



existing stocks of the compound could be used in certain



specified situations until December 31, 1986.  Formulations



unsuitable for conversion to uses in any of the specified



situations were discontinued after December 31, 1983.



Toxaphene is no longer commercially available (Berg, 1986).








     Once released into the environment, toxaphene is very



persistent in both soil and surface waters.  Toxaphene is



relatively insoluble in water and binds readily with soil,



consequently it is fairly resistant to leaching from the soil



column.  Biodegradation is not a factor in removing toxaphene
                             IV-2

-------
from aerobic soils, with reported half-lives of up to 20



years.  However, biodegradation in anaerobic soils can remove



up to 50 percent of the compound in 6 weeks.  Volatilization is



the only important removal process for toxaphene in shallow



soils.  The persistence of low levels of toxaphene in surface



waters over several years suggests that volatilization,



biodegradation, hydrolysis, and adsorption to sediments are



not rapid removal processes (Callahan et al., 1979).  Toxaphene



is a mixture of a number of chlorinated compounds; most of



these compounds are expected to bioaccumulate.








B.  Occurrence








Drinking Water








     Under the interim drinking water regulations, all



community drinking water suppliers are required to monitor



for toxaphene.  Since the regulations took effect, no system



has reported a violation of the 5 ug/L interim standard (FRDS,



1984).








     The 1978 National Rural Water Survey, as well as several



regional surveys, provide information on the occurrence of



toxaphene in public drinking water supplies.  According to
                             IV-3

-------
available information/ toxaphene concentrations did not



exceed the minimum quantification limit of 0.17 ug/L in the



71 groundwater systems tested (U.S. EPA, 1984).  Regional



studies of toxaphene in the 1970's have generally reported



that toxaphene, when it was detected, did not occur at levels



greater than 0.1 ug/L (U.S. EPA, 1987).  Current toxaphene



concentrations in drinking water supplies are believed to be



even lower than in the 1970's, due to the restrictions placed



on the use of the compound in 1982 and its later removal from



production.
Diet
     Although toxaphene is no longer used, the pesticide is



still expected to occur at low levels in many foods because



of its past widespread use and persistence in the environment.



Toxaphene has been detected in the FDA market basket surveys



(FDA, 1986).  Surveys conducted between 1982-1985 reported



finding daily intakes of toxaphene for 25-30 year old males



and females of 0.576 and 0.369 ug/day, respectively.  Dietary



levels are expected to decrease in the future because of the



discontinuation of the use of toxaphene.
                            IV-4

-------
Air
     According to U.S. EPA (1980) and Arthur et a)..  (1976),



particularly high toxaphene concentrations were reported,  in



several agricultural regions of the southern United  States in



the early and mid 1970's, with levels ranging up to  1.747 ug/m



The highest reported toxaphene level was 8.7 ug/m^ in Arkansas



in 1970 (Kutz et al., 1976).   In the 1970's, toxaphene was



widely used as an agricultural insecticide, and the  data from



the 1970's reflect this use.   Due to the discontinuation of



toxaphene in the last 5 years, ambient air levels today are



likely to be very low.








C.  Exposure Estimates








     The following table summarizes the current exposure



levels of toxaphene in drinking water, food and air.  The



drinking water levels are based on the range of the  levels



found in the regional studies in the 1970's.  The estimated



daily intake levels were made based on the assumption of an



intake of 2 liters per day of drinking water.  Current levels



are expected to be lower.  The dietary exposures are taken



from an average of several total diet studies taken  from



different regions of the United States from 1982 to  1985.
                          IV-5

-------
This number is a reasonable estimate of the average dietary

intake of adult males between the ages of 25-30 years old.

Some individuals would be expected to have a higher intake

than the average.

                             IV-5

              Exposure Estimates for Toxaphene
                      Reported Exposure
                      Levels (low-high)
                        Est imated
                       Adult Intake
Drinking Water

Diet

Air
0-<0.1 ug/L
Negligible
0-<0.2 ug/day

 0.6 ug/day

 Negligible
     The current available information on occurrence of

toxaphene is insufficient to determine the national distribution

of intake by any of the three routes.  However, EPA believes

that intakes from diet will generally be less than 0.6 ug/day

and that air exposure to toxaphene is expected to be negligible.

If toxaphene does occur in drinking water at levels of more

than a few tenths of a ug/L, it is likely to be the major

source of intake.  However, for the majority of individuals

in the United States diet is the major source.
                           IV-6

-------
D.  REFERENCES
Arthur, R.D., J.D. Cain, and B.F. Barrentine.  1976.
     Atmospheric levels of pesticides in the Mississippi
     delta.  Bull. Environ. Contain. Toxicol. 15 (2 ): 129-134 .

Berg, G.L. (ed.).  1986.  Farm chemicals handbook.  Meister
     Publishing Co., Willoughby, OH.

Callahan,  M., et al.  1979.  Water-related environmental
     fate of 129 priority pollutants.  Final Report.  EPA-
     44014-79129a.  Office of Water Planning and Standards.
     U.S.   Environmental Protection Agency, Washington, D.C.

FDA.  1986.  Food and Drug Administration.  Memorandum from
     E. Gunderson, Division of Contaminants Chemistry, Center
     for Food Safety and Applied Nutrition, Washington, D.C.
     to Dr. Paul S. Price, Office of Drinking Water, U.S.
     Environmental Protection Agency,  November 6, 1986.
     Washington, D.C.

FRDS.  1984.  Federal Reporting Data System.  Computer
     printout, dated April 4, 1984, containing data on organic
     chemical MCL violations, FY 1979-1983.  U.S. Environmental
     Protection Agency, Washington, D.C.

Kutz, F.W., A.R. Yobs, and H.S.C. Yang.  1976.  National
     pesticide monitoring programs.  In:  R.E. Lee (ed.).
     Air Pollution from Pesticides and Agricultural Processes.
     CRC Press, pp. 95-136, Cleveland, OH.

USDA.  1983.  U.S. Department of Agriculture.  Inputs.
     Outlook and situation.  Washington, DC:  Economic Research
     Service, U.S. Department of Agriculture.  IOS-2.

U.S. EPA.   1977.  U.S. Environmental Protection Agency.
     Toxaphene:  Position Document 1.  Washington, D.C.
     Special Pesticide Review Division, U.S. Environmental
     Protection Agency.  EPA/SPRD-80/55. Washington, D.C,

U.S. EPA.   1980.  U.S. Environmental Protection Agency.
     Ambient water quality criteria for toxaphene.
     EPA-440/5-80-076. Office of Water Regulations and
     Standards, U.S.  Environmental Protection Agency.
     Washington, D.C.
                             IV-7

-------
U.S. EPA.  1982.  U.S. Environmental Protection Agency.
     Decision document on toxaphene.  Office of Pesticide
     Programs, U.S. Environmental Protection Agency,
     Washington, D.C.  November 29, 1982.  47 FR 53784.

U.S. EPA.  1984.  U.S. Environmental Protection Agency.
     Rural water survey.  Computer data provided by
     Department of Sociology, Cornell University, Ithaca,
     NY.

U.S. EPA.  1987.  U.S. Environmental Protection Agency.
     Occurrence of pesticides in drinking water, food and
     air.  Office of Drinking Water, U.S. Environmental
     Protection Agency, Washington, D.C.
                              IV-8

-------
                         V.   HEALTH EFFECTS  IN  ANIMALS
Acute and Subchronlc Toxldtv
    Current  studies  of acute toxldty resulting from exposure  of  experimen-
tal animals  to toxaphene can  be  divided Into two main groups:  exposure  to
unfractlonated  (technical  grade)   toxaphene  and exposure  to specific  toxa-
phene components  or  subfractlons,  e.g.,  Toxicants  A and 8.   Tables  V-l,  V-2
and V-3 summarize  the L0c0  values  observed  1n each  of   these  experimental
situations.   Greater  than  10-fold  differences 1n  toxldty  have been  docu-
mented  for various  toxaphene  fractions or components that  differed  from each
other 1n chemical  composition,  polarity  and  solubility  (Pollock  and  KHgore,
1978a,b).

    Information  on the  acute  oral toxldty  of  unfractlonated  toxaphene  to
laboratory animals  1s  summarized  In Table V-l.  In cases  of acute  Intoxica-
tion,  toxaphene  like most  chlorinated  hydrocarbon Insecticides, appears  to
act as  a CMS  stimulant.   However, unlike  DOT,  toxaphene  does  not  signifi-
cantly affect conduction 1n  the rat superior  cervical ganglion  (WhHcomb  and
Santoludto,  1976).  Effects  of toxic exposures  In humans  (hypersens1t1v1ty,
tremors  and convulsions) are  similar to  those  observed  In  both  rats  and dogs
(Lehman, 1951).   Along with  convulsions, hyperreflexla  has  also been  noted
In dogs  (Lackey, 1949a), and rats  (Boyd  and  Taylor, 1971).

    It  Is  Important  to note that  humans who  Ingest a protein-deficient diet
may represent  a susceptible  population.  Rats fed a protein-deficient  diet
were more  susceptible  to toxaphene poisoning than rats fed  laboratory  chow,
LD5Q,  80+19  and  220*33 mg/kg  bw, respectively   (Boyd  and  Taylor,  1971).
02030                               V-l                              02/14/85

-------
o
co
o
                                                     TABLE  V-l

                Acute Oral  ToxicIty of Technical Toxaphene  (CAS RN 8001-35-2)  to  Laboratory Namnals
            Species
       Vehicle
                                                              (mg/kg bw)
                            Reference
   Rats:
     Unspecified strain
     Wtstar. male,
     (3-4 weeks. 50-60 g)
     low protein diet
     optimal diet

     Sherman, male.
     (>90 days. >175 g)

     Sherman, female.
     (>90 days. >200 g)
     Sprague-Dawley. male
     (24 days. 70 g)

     Unspecified strain.
     male
o
ro
CD
en
   Mite
corn oil
cottonseed oil
peanut oil


peanut oil

peanut oil

corn oil

olive oil (l.p.)


unspecified


corn oil


unspecified oil
   60
  220 t 33a
   80 * 19
  293 *• 31a
Lehman. 1948; Boots Hercules
Agrochemlcals. Inc.. n.d.
Boyd and Taylor. 1971
90(67-122)b      Galnes.  1960


 80(70-91)b      Galnes.  1960

   40            Shelanskl  and Gellhorn.  n.d.

  120-125        Shelanskl  and Gellhorn.  n.d.

  228            Pollock  et al..  1983


  270            Kuz'mlnskaya et  al.. 1980
  112            Boots  Hercules  Agrochemlcals.
                 Inc..  n.d.

   80            Rico.  1961

-------
                                                  TABLE V-l  (cont.)
o
ro
o
CO
Species Vehicle
Nice
Swlss-Uebstci , male corn oil
Cats peanut oil
unspecified oil
Dogs corn oil
kerosene
peanut oil
deodorized kerosene
< Rabbits peanut oil
^ deodorized kerosene
Guinea pigs corn oil
kerosene
unspecified oil
Hamsters:
Syrian Golden, male unspecified

L050
(rag/kg bw)

112
25 40C
100
20-25
>400
>30C
>450C
75-100c
250-500C
270
365
80

200

Reference

Fattah and Crowder. 1980
Treon et al.. 1950
Rico. 1961
Lackey. 1949a
Lackey. 1949a
Treon et al., 1950
Treon et al.. 1950
Treon et al.. 1950
Treon et al.. 1950
Boots Hercules Agrochemtcals,
Inc.. n.d.
Boots Hercules Agrochenilcals.
Inc.. n.d.
Rico. 1961

Cabral et al.. 1979
rvi
er
GO
       and  female.  6 weeks

     Calves:
       Mixed-breed.  136-232 kg
60X solution,
unspecified. (Cooper
Tox Emulslftable
Concentrate) diluted
with 2 l water
62
Steele et al.. 1980

-------
                                                  TABU V-l  (cont.)
IV)
o
Species
Cattle
Sheep
Goat
Horse
Monkey: female
Pheasant:
Unspecified strain
Mallard:
Anas platyrhynchos
Hen:
Unspecified strain
Vehicle
grain
xylcne
xylene
unspecified
peanut oil saline
(l-v.)

unspecified

unspecified

unspecified
L°50
(nig/kg bw)
144
200
200
80C
7.5-10

40

71

100
Reference
Boots Hercules Agrochemlcals.
Inc., n . tl .
Boots Hercules Ayrochemlcals,
I nc . , n . d .
Boots Hercules Agrochenitcals.
Inc., n.d.
Rico. 1961
Treon et al.. 1950

Netcalf. 1981

Hudson et al.. 1972

Rico. 1961
     aStandard error of the mean
     b95X confidence Interval
     cN1n1mum lethal dose
oo
-j

-------
                                  TABLE V-2

         Acute Dermal 1050 Values for Toxaphene In Laboratory Mammals
Species
Rats
Sherman, male
(>90 days, >175 g)
unfasted
Sherman, female,
(>90 days, >175 g)
Rats
Rats, male
Rabbits
Vehicle
xylene
xylene
xylene
unspecified
dust
L050
(mg/kg bw)
1075
(717-1613)*
780
(600-1014)*
930
940
-4000
Reference
Galnes, 1960,
1969
Galnes, 1960,
1969
Boots Hercules
Agrochemlcals,
Inc., n.d.
Ku?'m1nskaya
et al., 1980
Boots Hercules
Rabbits
peanut oil
-250
Agrochemlcals,
Inc., n.d.

Boots Hercules
Agrochemlcals,
Inc., n.d.
*95X confidence Interval
02030
         V-5
                   02/12/85

-------
o
Gi
O
                                                      TABLE V-3

               Acute Toxlctty of Technical Toxaphene Components Administered IntraperHoneally to Ntce
        Toxaphene  Component
             (CAS  RN)
Species/Strain
Vehicle
                                                                     1050
                                                                  (mg/kg bw)
Reference
i
0>
2.?.5-endo-6-exo-8.
9,10-heptachloro-
bornane
(51775-36-1)

2.2.S-endo-6-exo-8.
9,10-heptachloro-
bornane
(51775 36-1)

2.2.S-endo-6-exo-8.
9,9,10-octachloro-
bornane
(58002-19-0)

Technical toxaphene
                                  mice/Albino
                                  male
                                  mice/
                                  unspecified
                                  mice/
                                  unspecified
                                  mlce/SwIss-
                                  Uebster. male

                                  rats/Sprague-
                                  Dawley. male
                    unspecified
                    unspecified
                    unspecified
                    dimethyl
                    sulfoxlde

                    olive oil
                    75        Saleh  et  al..  1977
                     6.6       Khalifa  et  al..  1974
                     3.1       Khalifa  et  al..  1974
              33(27-41)*       Pollock  and  Kllgore. 1980
                   228        Pollock  et al..  1983
     *95X confidence  limits
o<
V.
00

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Clinical  signs  of  depression  and stimulation  of  the CNS  were  the  same  In
both groups;  however,  signs  appeared earlier and at  lower  concentrations  1n
protein-deficient  rats.   Pathological effects  Included  cloudy  swelling  and
congestion of the  kidneys, fatty  degeneration  and  necrosis  of  the  liver,  and
decreased  spermatogenesls.   Mehendale  (1978)   reported  that  toxaphene  (100
ppm In the diet for 8 days) Inhibited hepatoblHary function In rats.

    The  acute  dermal  toxldty  of  toxaphene   Is  summarized  1n  Table  V-2.
Toxaphene  appears  to  be  somewhat  less  toxic when  administered  dermally
although  toxldty  appears to  be  carrier-dependent.   In rats  the ratio  of
dermal  to  oral  LD5Qs   1s  12  (1075/90)  for   males   and  9.8  (780/80)  for
females  (Galnes,  1960,   1969).   Dermal  LDc-s  for  rats  range  from  780-1075
mg/kg bw (Galnes, 1960,  1969;  Boots Hercules Agrochemlcals,  Inc., n.d.).

    A  40% technical  toxaphene  dusi  (3-4  g/ma)  killed  around half of  an
exposed  group  of  rats   after  1  hour  (Boots  Hercules Agrochemlcals,  Inc.,
n.d.).   Assuming  that  a 200 g  rat  Inhales  200 ml  of  air  per minute  and
absorbs  100% of  the  Inhaled dose,  a 3 g/m3   concentration  for 60  minutes
would equal 72 mg/kg bw.

    Repeated  (14 applications) administration  of a 20% solution  of  toxaphene
1n  kerosene   to  the eyes  of rabbits  (0.01 ml)  and  guinea  pigs   (0.05  ml)
caused mild   Irritation  to  the eyelids  with  loss  of hair  of  the  eyelids.
There was  no Injury to  the eye and  condition  on the  Hds  cleared  completely
In 10 days (Boots Hercules Agrochemlcals, Inc., n.d.).
02030                               V-7                              02/14/85

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    Hyde  et al.  (1978)  compared  the  acute electrophyslologlcal  effects  of
representative  organochloMne  and  organophosphate Insecticides on laboratory
rats.   Concurrent  neurological,   respiratory  and  cardiac   Involvement  were
primary  concerns.   Each  of   14  male  Sprague-Dawley  rats   (240-280 g)  was
equipped  with  dural  electrodes   to  record   spontaneous  cortical  electro-
encephalograms.   The  animals  were  given a minimum  of  7 days to recover from
electrode  Implantation  and  were  randomly  assigned  to  1   of  7  treatment
groups,  2  rats/group.   Treatments consisted  of  separate  lethal  1.p.  Injec-
tions  of each  of  three organochlorlne  pesticides,   Including  70 mg  toxa-
phene/kg  bw and  three  organophosphates  emulsified  1n a Tween-80  1soton1c
saline  solution.    Controls  received  0.5  ml  of  the  vehicle only.   Total
percentage changes  1n  cardiac  and  respiratory  rates during exposure  to toxa-
phene  (expressed  as  percentage change  from  pretreatment to late stage  of
toxlclty) were  ECG rates,  +-17.5 and respiration,  +56.5.  The distortion  of
spontaneous  cortical  potentials recorded during  toxaphene  exposure  revealed
a cerebral sensitivity.

    Toxaphene  1s  used extensively  for  external parasite  control  In  domestic
animals.   The  maximal  nontoxlc   concentration  for  topical  application  In
adult  swine 1s a 1.5X  solution.   01P1etro and  Hallburton  (1979)  reported
signs  of  a  CNS disorder In  about 40  of a herd  of around  150  feeder  pigs.
Thirty-six hours  earlier  the  affected  animals  had been treated for sarcoptlc
mange  with  a   toxaphene  mixture  applied at  -10  times   the manufacturer's
labeled  dosage   (300  ml  of   61%  toxaphene  stock  In  4 i   H?0).   One  day
after  the  treated pigs were sprayed with warm water,  severely affected pigs
had Improved markedly.   By day  5 the animals appeared  normal.
02030                               V-8                              02/14/85

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    Deaths and abnormal behavior  In  a  herd  of  50  crossbred  beef  cows  In  good
body  condition  were  attributed  to  an  Ingestlon of  toxaphene-contamlnated
feed  (Braun  et  al.,   1980).   Levels  of  11-35  mg  toxaphene/kg  hay  were
detected 1n four samples from pastures where the animals  grazed.

    With  the  objective of  determining  tissue  levels  of  toxaphene  following
exposure  to  a single  oral  dose of  toxaphene,  Steele  et al.  (1980)  exposed
heifer  calves  ranging   1n weight  from 136-232 kg  to  the following doses  In
mg/kg bw  (number of animals  exposed  to  each dose 1n parentheses):   50  (6),
100  (7),  150  (6).   Toxaphene  originally available  as  a  61% solution  was
diluted  with   -2 l  of  water.  Surviving  animals were   sacrificed  after  7
days.   In  the low dosage group, there were  2/6  deaths  after 4 days;  In  the
100 mg/kg  exposed group  there  were  6/7 deaths within  4.5-42 hours, and  In
the high dose  group, there  were 5/6  deaths  within 21  hours  to 5  days.   Clin-
ical signs of  toxIcHy  Included apprehension and  hyperexcHabUHy,  followed
by  anterior,   then  posterior, muscle  faslculatlons.   Terminal  stages  were
characterized  by  generalized  muscle  twitching and clon1c-ton1c  convulsions.
When  toxaphene tissue  levels  were  determined,  1t  was  observed  that  while
kidney and brain  toxaphene  levels  were not  dlagnostkally significant,  liver
residues  of  toxaphene  >4  mg/kg ww  were associated with lethality for  the
7-day observation period  (Steele  et al.,  1980).   By  contrast, brain  levels
of toxaphene may be Indicative of  acute  toxldty; as  a  result of  Inadvertent
dermal exposure  In swine, >4  mg/kg ww  In the brain was  found to  constitute a
lethal  level,  and  2  mg/kg was  associated with  clinical  signs  of  toxldty
(OlPletro and  Hallburton, 1979).  A  summary  Is  provided  In  Table  V-4.   Acute
Intoxication of horses  has  also been reported  following  Ingestlon  of  alfalfa
contaminated with DDT and toxaphene  (level  not  reported)  (Nazarlo  and  Capel-
laro, 1980).
02030                               V-9                              02/14/85

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                                   TABLE V-4

               Residue Concentrations 1n Dead Birds  and  Animals
         Animal
                             Organ     Residue Range3
                                         (mg/kg ww)
                         Reference
Female bat,  Tadarlda
brasH1ens1s Hexlcana)
Rabbit (2/31 )b
Deer (3/22)b


Heifer calves (6/7)b
Swine
                            carcass
                            carcass
                            carcass
                            liver
                            brain
1.2-12.4
1.7-8.7
4 mg/kg was
critical level for
acute exposure

4 mg/kg was
critical level for
acute exposure
                      Geluso et  al.,
                      1981
Causey et al.,
1972

Causey et al.,
1972

Steele et al.,
1980
DIPIetro and
Hallburton, 1979
Bald eagle (8/49)b
( 20/49 )b
(7/50)b
(ll/50)b
(13/69)b
(22/69)b
Great blue heron (9/35)
(6/36)b
Cattle egret (3/3)b
(2/3)b
Great egret (1/1 )b
(1/1 )b
brain
carcass
brain
carcass
brain
carcass
brain
carcass
brain
carcass
brain
carcass
0.13-1.2 (1975) Kaiser et al.,
0.06-2.5 (1975) 1980
0.05-0.31 (1976)
0.05-0.55 (1976)
0.06-2.7 (1977)
0.06-0.88 (1977)
0.17-0.82 Ohlendorf
(1972-1978) et al., 1981
0.11-0.50
(1972-1978)
0.28-0.36 (1978)
0.11-0.16 (1978)
0.54 (1978)
0.58 (1978)
aRange In positives

bPos1t1ves 1n total birds and animals examined
02030
                                    V-10
                              02/14/85

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    The  acute  toxldty of  toxaphene  components In mite  Is listed  In  Table
V-3.  In male Sprague-Dawley 24-day-old rats,  the  less  polar  toxaphene  frac-
tion, Rf 0.35-0.85, and  a more  polar  fraction,  R   0-0.57,  1s Included  In
this  table  and  exhibited  LD5Q  values of 217  and  266 mg/kg bw, respective-
ly,  compared with   toxaphene,  R,  0-0.85,  LD5Q  228  mg/kg.   The  route  of
administration was  1.p.,  and  the  solvent  used  1n  the  toxldty  determination
was olive oil (Pollock et al.,  1983).

    In a  comparative study certain polychlorobornanes  that  are known  to  be
components of toxaphene  have  been examined for  structure-toxldty  relation-
ships (Saleh et al., 1977;  Saleh  and  Caslda,  1979);  using  2,2,5-endo, 6-exo.
8,9,10-heptachlorobornane  as  a  standard with a  relative  toxldty  of  100
(LDcQ 1.p.  75 mg/kg bw  1n male  albino mice).  An  enhanced acute  toxldty
was associated with the  Introduction of  a  single  chloro group In  the  8- or
9-pos1t1on, and  to  a lesser extent  In  the  5-exo-position.  Pretreatment  of
mice with  plperonyl butoxlde  (which  modifies  cytochrome P-450-med1ated  oxl-
datlve  or  reductive detoxification  mechanisms)  was  associated  with  lower
acute LD5Q  values  (2- to 8-fold)  for  the heptachlorobornane compound and  a
mixture  of  Its  8-  or 9-monochloro  derivatives.   Acute toxldty was  reduced
relative to  the  standard  when  monochloro groups  were Introduced   1n either
the 3-exo or  I0-pos1t1ons,  but  the toxldty of  the  10-chloro  derivative  was
also enhanced 2- to 8-fold by  pretreatment  of mice with plperonyl  butoxlde.
Dehydrochlorlnatlon  of  the hexachlorobornenes  with  apparent removal of  the
6-chloro  group   yielded   a  hexachlorobornane  with only  slightly  Increased
acute toxldty relative to the parent compound.
02030                               V-ll                             02/14/85

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    LD5Q  values  for  20-29 g  male  Swiss-Webster   mice  were  compared  for
toxaphene and  for  three  toxaphene fractions  dissolved 1n  dimethyl sulfoxlde,
and  administered  1.p.   Only  one  fraction,  with  an  L05Q of  20 mg/kg  bw
(19-21, 95X  confidence  Intervals),  was more  toxic  than  toxaphene (33 mg/kg)
Itself (Pollock and Kllgore, 1980).

    Table V-5  summarizes the effects  of  acute and  subchronlc  oral  adminis-
tration of toxaphene  to  laboratory  mammals.   Except for convulsions  observed
1n dogs given  10 mg/kg bw/day  (Lackey,  1949a),  few of the exposures  detailed
In Table V-5 resulted 1n clinical signs of toxaphene poisoning.

    Lackey (1949a)  exposed  dogs of an unreported breed and sex both acutely
and subchronlcally  to  toxaphene.   Dosing was performed by  stomach tube with
the toxaphene  dissolved  1n  corn  oil  or  kerosene  for  acute  experiments  and
corn oil administered as a  single dose dally 1n a gelatin capsule for longer
term  exposures.   None of  the  three  animals  receiving a  single dose of  5
mg/kg bw 1n  corn oil  developed  convulsions  as compared with 4/5 treated with
10 mg/kg.  At  doses >15  mg/kg  the convulsions were accompanied by Increasing
mortality.    Of  those treated  with  toxaphene  dissolved  1n   kerosene,  the
minimum  dose  at   which  animals  had  convulsions  was  75  mg/kg; none  were
recorded for two  dogs  dosed with  25 mg/kg  or  one dog  receiving 50 mg/kg.
These effects may actually Indicate a possible vehicle effect.

    Dally treatment with 5  mg/kg toxaphene resulted  1n  the onset of convul-
sions  after  a  few  days.  For  subchronlc treatment  the  dosage was therefore
decreased to 4 mg/kg bw.  Two  dogs  were treated dally at  this level  for  44
days  and  two  for  106 days.   One  dog treated  for 106  days   and both  dogs


02030                               V-12                             02/14/85

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                                                                             TABLE V 5



                                                          Acute and Subchronlc Oral Toxtctty of Toxaphene
C3 	
CO
O
Species
Nice, albino and
wild strains
Albino rats
Rats
Rats
Rats. SherMan.
* Mile and feMale.
- 100,
Rats
Rats. Osborne-
Nendel
Rats. Osborne-
Nendel
NuMber of Vehicle
AnlMals
5-18 diet
12N. 12F diet
at each
level
12N. 12F diet
unknown NS
6N. 6F diet
at each
level
4 diet
SN. SF acetone
at each
level
SN. SF acetone
at each
level
Duration
Several weeks
or Months
4. 8 and 12
weeks •
12 weeks
7 Months
2-9 Months
8 days
6 weeks
6 weeks
Dose
(Mg/kg/day or
ppM In the diet)
SO Mg/kg/day
(250-480)*
2.33
7.0
21.0
63.0
189.0
189*
1.2-4.8 Mg/kg/day
SO and 200*
100*
160*
320*
640*
1280*
2S60*
1280*
2S60*
SI 20*
Response
Changes In blood cheMlstry
and urine protein
No effect on weight.
liver cell histology
No apparent adverse effects
Temporary change In blood
cheMlstry
Questionable liver pathology
Decreased bile production
No deaths and Mean weight gains
unaffected (1280). 2 deaths
(both F) and Mean weight gains
of survivors not adversely
affected (2S60)
Two deaths (IN. IF) at 2S60
Reference
BluMler. 197S
Clapp et al. ,
1971
Clapp et al..
1971
Crebenyuk. 1970
Ortega et al..
19S7
Hehendale. 1978
NCI. 1979
NCI. 1979
CO

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                                                                          1ABU  tf-S  (cont.)
CJ
O
Species
Nice. B6C3F1
Dogs


Nuaber of Vehicle
AnlMls
5H. 5F acetone
»t each
level
3 corn oil
S corn oil
2 corn oil
Duration
6 weeks
1 exposure
1 exposure
44 days
Dose
(•g/kg/day or
PPM In the diet)
40*
80*
160*
320*
640*
1280*
S wi/kg/day
10 ag/kg/day
4 Mg/kg/day
Response
? deaths (1 H. 1 F) and mean
weight gains unaffected (320).
6 deaths (4 H. 2 f ) (640)
No convulsions
Convulsions In 4/5 animals
Questionable liver pathology:
Reference
NCI. 1979
Lackey. 1949a
Lackey. It49a
Lackey. 1949a
                                                corn oil
106 days
4 ag/kg/day
renal tubular degeneration;
sporadic convulsions

Questionable liver pathology:
renal tubular degeneration;
sporadic convulsions
Lackey. 1949a
           •pp» toxaphene In the diet
           NS > not specified; N - male; f - feaale
 CO

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 treated  for  44 days lost weight  during  the  course of exposure.  Convulsions
 were  reported  to  occur  tfnly  occasionally.   There were no changes In complete
 blood  counts  throughout  the  experiment  and  no gross  pathological  changes
 were  found.   Upon  hlstopathology  the  liver  showed   generalized  hydropic
 degenerative  changes,  which  appeared  to  be  reversible,  and  degenerative
 changes  In the parenchyma.

    In  both  the  Lackey  (1949a)  study  (using  dogs)  and  the  Ortega  et  al.
 (1957)  study  (using  rats)  changes  1n  liver  histology were  noted  1n  the
 animals  dosed  with  toxaphene.   Morphologically,  these  changes  appeared  as
 vacuoles of  plasma  with occasional red blood  cells  appearing  within hepatic
 cells.   This condition,  referred to as  hydropic accumulation,  Is  distinct
 from  fatty degeneration.  In  neither  rats  nor  dogs was hydropic accumulation
 associated with the destruction  of hepatic  cells.  Ortega et  al. (1957) also
 noted occasional  masses of  red  blood cells Invading  the  cytoplasm  of liver
cells   In  areas   of   centrolobular-cell   hypertrophy  and  marg1nat1on  was
 observed In  50% of  the  rats  fed  a diet  containing 200 ppm toxaphene from 2-9
 months.  Similarly, 27% of rats  fed  50  ppm showed this  same effect.   No con-
 trol rats were examined however.

    In  addition  to  liver  damage,  Lackey   (1949a)  documented  widespread
 degeneration of  the  tubular epithelium of  the  kidney,  occasionally  accom-
 panied  by  Inflammation  of  the  pelvis  of  the  kidney  for  dogs  fed  4 mg/kg
 bw/day  for 44  and 106  days.   Similar pathological  changes  were seen In dogs
 and rabbits  who survived  prolonged dermal  exposures  [200-600  mg/kg bw for up
 to  32  days  (dogs); 100-1000 mg/kg bw for  up  to  30  days (rabbits)]  to toxa-
 phene  (Lackey,  1949b).   For  dogs, the lowest lethal dose was  600  mg toxa-


 02030                               V-15                             02/26/87

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phene/kg  bw;  for  rabbits,  the lowest  lethal  dose  was  600  mg/kg bw when dis-
solved  In mineral  oil  or dimethyl  phthalate,  with  the highest, nonlethal dose
being  500 mg/kg  bw when applied  as a  dust  for  5-14  applications  {Lackey,
1949b).   Ortega  et  al.  (1957),  however,  did not   note  any  pathological
changes attributable  to  toxaphene  1n the  kidneys  of  rats  fed  a 50 or  200 mg
toxaphene/kg diet  for up to 9 months.

    Allen et  al.  (1983)  studied  the  effects  of  Ingested toxaphene  on  the
Immune  response of  female  Swiss-Webster  mice.   A total  of  130  8-week-old
mice were given  feed  containing 10,  100  or  200  ppm toxaphene for  8 weeks
(acetone  solvent).   Humoral  antibody production, IgG  antibody formation,  was
suppressed  1n  mice receiving  the  100  and  200  ppm doses.   Cell-mediated
Immune responses were not affected by toxaphene  treatment.

    Clapp et  al.   (1971)  fed  technical  grade  toxaphene to  groups  of  12 male
and 12  female  albino rats (144  total) 1n  their diets for   4,  8- and  12 weeks
at levels of  0, 2.33, 7, 21, 63 and 189 ppm.   Four  animals  from  each group
were sacrificed  at each  time  Interval.   Two  male rats died  during  the test
period  but  their  deaths  did  not appear  to be  caused  by toxaphene levels In
the diets.  Toxaphene  had no measurable adverse effects on physical  appear-
ance,   gross  pathology, weight  gain or  liver cell histology  of  any  of  the
experimental animals.

    Cattle and  sheep have  been  fed  toxaphene  at  concentrations  as  high as
320 ppm In  hay  for 134-151  days.   Central  nervous  stimulation with muscle
tremors occurred  In  steers  at the  highest  dose, but not In sheep  receiving a
similar  dose.   There  appeared  to  be no   abnormal  hlstopathology or  blood
chemistry (Boots Hercules Agrochemlcals,  Inc., n.d.).

02030                               V-16                             02/26/87

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    Guinea  pigs  suffered 73X  mortality when  solutions  of 20%  toxaphene  \n
mineral oil,  equivalent  to  a dose of  332  mg toxaphene/kg bw/day,  was admin-
istered for 14 days (Boots Hercules Agrochemlcals, Inc., n.d.).

    Feeding  studies  were conducted  to estimate  the maximum  tolerated doses
of  toxaphene  In Osborne-Mendel  rats  and  B6C3F1  mice  for a  National Cancer
Institute  study  of cardnogenlclty (NCI,  1979).  Toxaphene was  dissolved  In
acetone and  added  to the feed.   Corn  oil  was also added  as  a dust suppres-
sant  1n  an  amount  equal  to  2%  of  the  final   weight  of feed.   Treatment
groups, consisting of  5  male and  5  female animals  1n  each group,  were given
food  with or  without toxaphene   for  6 weeks,  and they  were  observed  for
another 2 weeks.

    For rats,  toxaphene  was added  to  the  feed  In  2-fold Increasing concen-
trations,   ranging  from 160-2560  ppm  feed.  A second  study was  performed  on
male and  female  rats  at  toxaphene levels  ranging from 1280-5120 ppm to con-
firm the results and to extend the concentration  range of the  first study.

    At  1280 ppm  1n the first and  second  studies, there  were  no deaths among
rats and  mean weight  gains  of both  sexes were  comparable to controls  (see
Table  V-5).   This  Is a  NOAEL  for mortality  over a 6-week period.   At  2560
ppm, two  female  rats  died In the  first study but mean weights of the surviv-
ors were  not  adversely affected.   During  the  second  study,  one male and one
female  died  at  2560 ppm.   On  the basis  of these results, the  low and high
doses  for  the chronic studies were  set at 1280  and 2560  ppm for  males, and
640 and 1280 ppm for females.
02030                               V-17                             02/26/87

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    In  the  same 1979 NCI  subchronlc  study, mice were  given  feed containing
40-1280  ppm  toxaphene 1n_  the  diet.   Four  males and two  females  died  at  640
ppm,  and  one male and one  female  given  320 ppm died.  Mean  weight  gains  of
mice  given 320  ppm were comparable with those  of controls.   On  the  basis  of
these results the  low and  high doses  for  the chronic studies were set at  160
and 320  ppm  for males and  females, respectively.   The  160  ppm diet  dose  can
be  regarded  as  the  NOAEL   In  mice  with  respect  to mortality over  a  6-week
period.

    Toxaphene aerosols  1n  the  form of dusts are more  toxic  to  rats  than  In
the  form of  mists.   M1st  concentrations  as  high as  500  mg  toxaphene/m3
caused no mortality In rats  and  rabbits  over a 3-week period.   However,  no
rats  survived dust concentrations of  250  mg/ma for 1  week.   Rats,  dogs  and
guinea  pigs  were  killed  at  12  but  not   at  4  mg dust/m3  over a  3-month
period  of exposure.   Some  surviving  female  rats  exhibited  slight  focal
hepatic cell  necrosis (Boots Hercules Agrochemlcals, Inc., n.d.).

Chronic ToxIcUy
    Long-term exposures to  low dietary levels  of toxaphene  are summarized  In
Table V-6.  All  studies  note  some form of  liver pathology  1n rats  at levels
>100  ppm  1n  the diet.   At  this same  dose,  cytoplasmlc  vacuollzatlon similar
to that  seen  following  subchronlc oral exposure was  noted  by Kennedy at  al.
(1973).  Lehman (1952) also reported  fatty  degeneration of  the liver 1n rats
fed  100  ppm.   With  a  25  ppm diet, Fltzhugh and  Nelson  (1951)  observed
Increased  liver weight  with  minimal liver  cell  enlargement.   Unpublished
studies  on  rats,  dogs and monkeys  by  Boots  Hercules  Agrochemlcals,  Inc.
(n.d.) are In general agreement with  the above published reports.

02030                               V-18                             02/14/85

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                                                          TABLE V-6
0
IVi
o
co
o
Chronic Toxlclty of
Duration
Species of Feeding
(number of animals)
Toxaphene to Laboratory
Toxaphene
Concentration
(ppm diet)
Rats. 3 generations 25
Sprague-Dawley (42 weeks for FQ
(64N. 128F. and 39 weeks for 100
weanling) Fj and F2>


to



o
Rats lifetime

Rats lifetime


Dogs 2 years
2 years
25
100
25
100
(20N. 20F)
1000-1500
(20M. 20F for
each dose)
5,10.20
200
(IF. IN)
Hammals at Low Dietary Levels
Response Reference
No effect Kennedy et al..
1973 |
Liver pathology
No effect Lehman. 1952
Liver pathology
Liver pathology Fltzhugh and
Nelson. 1951
No effects
Slight liver damage. CNS
stimulation
No effect on organ Boots Hercules
weights, histology Agrochemicals.
or clinical tests Inc.. n.d.
Moderate liver degen-
eration
03
in

-------
                                                   1ABLE  V  6  (cont.)
ru
O
Duration
Species of Feeding
Dogs (6) 1360 days
(3.7 years)
up to
1260 days
Toxaphene
Concentration
(ppro diet)
200* (4)
(5 mg/kg/day)
400* (2)
(10 mg/kg/day)
Response Reference
Liver necrosis Boots Hercules
AgrochemUals.
Inc.. n.d.
One death after 33 days
     Bobvhtte quail
     Collnus vlrglnlanus
     male
     Unite Leghorn
     Chicks, female
138-171 days
up to 50 weeks
10.0
                                                      50.0
 0.5
                                                       5.0



                                                      50.0

                                                     100.0
138 days SOX errors In
behavioral tests

171 days-same as
controls

Occasional deformation
of the cartilaginous
region of the keel.
Increased growth of
cartilage.

Sternal deformation and
mild nephrosls at 5. 50
and 100.

Renal lesions at 50. 10Q

Decreased body weight
Kreltzer. 1980
Bush et al.,
1977
S   'Administered In capsules containing toxaphene In corn oil
\
£   H -- male; F = female
\
03

-------
    Kuz'mlnskaya  and Ivan1tsk11  (1979,  1981)  showed  that  a single  dose to
rats,  one-half  of  the LD5Q  of  toxaphene (120  mg/kg  bw),  decreased  the
amount  of adrenaline  1n adrenal  glands  and  Increased adrenaline  and  nor-
adrenallne  1n  the  urine  during the  first  24 hours  after  administration of
the  pesticide.   A  chronic  study  was   Initiated   using  albino  male  rats
(200-250 g)  that  were  given dally doses  of  toxaphene (2.4  mg/kg bw) up  to 6
months.  The number  of  animals  per group  was not reported.   Chronic adminis-
tration altered  catecholamlne  metabolism  and breakdown.  There were disturb-
ances  1n  catecholamlne  metabolism In toxaphene  treated  animals  1n the brain
and the heart  that  may  account  for the clinically observed damage to the CNS
and the myocardium.

    The NCI  (1979)  conducted  a chronic  study  with  Osborne-Mendel  rats  and
B6C3F1 mice  to determine  the  possible carclnogenldty  of  toxaphene.   Toxa-
phene  was  added  to feed  as  described,.  1n  the  subchronlc  effects  section.
Fifty animals  of  each  gender constituted a  treatment group of  rats or mice.
Ten untreated  animals  of each gender were  matched  controls and  data from 45
                                         *
or 40  untreated  animals from  similar bloassays  were  pooled  for statistical
evaluations.

    Groups  of  50 mice/sex  were  administered   toxaphene  for 80 weeks  and
observed until  sacrifice at 90-91  weeks  (NCI,   1979)  (Table  V-7).  Low-dose
males and  females received 160  ppm  In  the  feed  for  19  weeks  followed by 80
ppm for 61 weeks;  high-dose  males  and females,  320 ppm  for 19 weeks, follow-
ed by  160 ppm for  61  weeks.  TWA doses  were  99 or  198 ppm  1n  the diet for
both males  and- females.  Mean  body  weights attained  by high-dose male  mice
were  lower  than  those  of  matched controls, but  weights  of  other  exposure


02030                               V-21                             02/26/87

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                                   TABLE  V-7

               National -Cancer Institute Chronic Feeding Study3
               Duration    Toxaphene   Time-Weighted
   Species    of Feeding Concentration Average Doseb
               (weeks)    (ppm diet)     (ppm diet)
                                         Response
  High-dose
Females

  Low-dose


  High-dose
19
61

19
61
19
61

19
61
Rats,
Osborne-Hendel
  High-dose
 2
53
25

 2
53
25
 160
  80

 320
 160
  99


 198
 160
  80

 320
 160
  99


 198
1280
 640
 320

2560
1280
 640
                                           556
1112
Mean body weights
unaffected

Mean body weights
adversely affected;
several animals died
before week 19; dose-
related decrease In
survival
Mean body weights
unaffected

Mean body weights
unaffected; several
animals died before
week 19; dose-related
decrease 1n survival
            Mean body weights
            unaffected
HyperactlvHy (week 2);
generalized body
tremors (week 53);
mean body weights
unaffected; no dose-
related decrease 1n
survival
02030
                   V-22
                                         02/26/87

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                               TABLE  V-7  (cont.)
               Duration    Toxaphene   Time-Weighted
   Species    of Feeding Concentration Average Dose''
               (weeks)    (ppm diet)     (ppm diet)
                                         Response
Females

  Low-dose


  High-dose
55
25

55
25
 640
 320

1280
 640
 540
1080
Mean body weights
adversely affected

Mean body weights
adversely affected;
generalized body
tremors (week 53); no
dose-related decrease
In survival
aSource: NCI, 1979

bT1me-weighted average dose
             Ifdose In ppm x no. of weeks at that dose)
                z(no.  of weeks receiving each dose)
02030
                   V-23
                                         02/26/87

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groups  were essentially  unaffected by  toxaphene.   Several animals  died In
the  high-dose  groups  before  week  19  when doses  were  lowered.   Following
this, the exposed  mice  were  generally  comparable with controls 1n appearance
and  behavior  during  the  first year of  the study.  During  the  second year,
abdominal dlstentlon  was  observed  1n  all  dosed groups  but  predominantly In
the  high-dose  males.  Other  clinical  signs  Included  alopecia,  diarrhea,
rough hair  coats  and  dyspnea.   From weeks  60-76 the low-dose males appeared
hyperexcltable.   After  week 75  there  were dose-related  differences  1n sur-
vival.  Because of  the  varied  dose  regimens,  valid NOAEL data cannot be cal-
culated.

    Groups  of  50  rats of  each  gender  were  administered  toxaphene  for  80
weeks  (NCI,  1979)  and  then  observed  until  survivors  were sacrificed  at
108-110 weeks  (see Table  V-7).   Low-dose  males  were  given 1280  ppm 1n the
feed  for  2  weeks,  640 ppm for 53 weeks  and 320 ppm for  25 weeks, which was
reported as a TWA  of  556  ppm.   High-dose males were administered 2560 ppm In
the feed  for  2  weeks, 1280 ppm  for 53 weeks and  640  ppm for 25 weeks, or a
TWA, of  1112 ppm.   Low-dose  females  received 640 ppm 1n the feed for 55 weeks
and 320 ppm for 25 weeks, for a  TWA of  540 ppm.  High-dose females received
1280  ppm  1n the feed  for  55 weeks, and  640  ppm for 25  weeks,  for  a TWA of
1080 ppm.

    Mean body weights of  the low-  and high-dose female rats were lower than
those of  the  matched  controls  throughout most  of  the bloassay study period,
whereas weights  of  low- and  high-dose  males  were  essentially unaffected.
During  the  first  16  weeks,  dosed males were  generally comparable with con-
trols  1n  appearance  and   behavior, with  the  exception  of  high-dose  males

02030                               V-24                             02/26/87

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 that  appeared  hyperactive  during  week 2 when  the  doses for male  rats  were
 reduced.  At week  53,  tn-e concentration  of  toxaphene In the feed was reduced
 because a majority of  the  high-dose males  and  females  developed generalized
 body  tremors.   Dose-related decreases  1n  survival  rates were  not  observed.
 Clinical signs  usually associated  with aging were  observed  earlier  In dosed
 rats  than  1n  controls and  were  as  follows:   alopecia, diarrhea,  dyspnea,
 pale  mucous  membranes, rough hair coats, dermatitis, ataxla,  leg paralysis,
 eplstaxls,   hematurla,   abdominal   dlstentlon   and   vaginal   bleeding.   Two
 females, one high-dose and one low-dose,  had  Impaired  equilibrium.   Again,
 no valid NOAEL data 1s available for this study.

    Bush et  al.  (1977) added graded  levels of toxaphene In corn  oil  to the
 diets of  female White Leghorn  chicks  from  1  day of age  In  order  to achieve
 doses of  0,  0.5,  5,   50  and 100 ppm  diet.   Each  treatment consisted  of  90
 randomly selected  birds'(30 birds  1n  each  of  three  replicates).   Levels  up
 to 100  ppm  diet did not  produce symptoms of  toxldty throughout the 50-week
 study.  Mortality  was <5X  In  all groups.   Body weights at 6  and  30 weeks
were  significantly decreased when  the  birds were fed 100 ppm.   There were no
 significant  treatment-related   changes  1n  heart,  liver,  gizzard or  kidney
weights at  4 or 8  weeks.  Necropsy of  31-week-old birds fed  5,  50 and 100
ppm  toxaphene  1n  the  diet  revealed sternal deformation  resembling  osteoma-
 lada.  Occasional  keel  deformation  Involving  the cartilaginous tissue  as
well  as an  apparent Increase 1n  growth  of  cartilage was found In  birds fed
0.5  ppm toxaphene  In  the  diet.   Further  studies  were  planned to determine
whether the  backbone  of  the chicken  as well  as  other  skeletal  structures
were  affected  by   toxaphene.   Hlstopathologlcal  examination  of  organs  of
 31-week-old  birds  showed  mild nephrosls  of  the kidney In birds  fed toxaphene


 02030                               V-25                             02/26/87

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at  5,  50 and  100 ppm.   Other  observations  Included cysts,  occasional  cel-
lular casts  In the renrt  tubules  and accumulation of a brown  granular  pig-
ment 1n the cytoplasm of some tubular epithelial cells.

    Kreltzer  (1980) determined  whether  behavioral effects  1n birds  would  be
produced  by  toxaphene  at  levels  below  those  that  produced  overt  signs  of
Intoxication.  Adult  male  bobwhlte quail Collnus  v1rq1n1anus were  fed  toxa-
phene dissolved  In  propylene glycol  and  blended Into the  feed at  levels  of
10 and 50 ppm.  An equal  amount  of propylene glycol  was  added to  the feed  of
controls.  There  were four controls  and  four  birds  per  treatment  level  For
each of  two  tests (with different pairs of  patterns)  that  measured perform-
ance  on  nonspatlal   discrimination  reversal  tasks.   Birds  were  Initially
exposed at 3  days of  age for 138 and 171 days  before tests 1 and  2, respec-
tively.   The  treated  birds 1n  test  1  had  SOX more errors  than the controls
(p<0.02).   There  were  no  significant   differences   In  the performances  of
birds fed the  two toxaphene levels.  In test 2,  after 171  days of exposure,
treated birds  performed  as  well  as  the  controls,  Indicating  an accomodatlon
to the chemical.

Enzyme Effects
    Toxaphene appears  to  Induce  the  mlcrosomal  MFO system as  evidence by the
shortening of  pentobarbHal  sleeping time  at 50  mg  toxaphene/kg  bw (Schwabe
and HendUng,  1967).   A  dose  of  100  mg toxaphene/kg bw  has been  shown  to
decrease  the barbiturate  sleeping  time  by  -20% for up  to  20   days  after
exposure (Ghazal, 1965).  Toxaphene  will also  Initiate MFO  activity 1n  whole
liver homogenates from  rats pretreated  with  toxaphene  (Klnoshlta  et  al.,
1966).  Effects  1n  male rats were more pronounced than  females.   Toxaphene

02030                               V-26                             02/26/87

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also appears  to  stimulate  liver  mlcrosomal  metabolism of  estrone and testos-
terone.   For  example, an  1.p.  dose  of  120 mg/kg  bw to  male rats  caused  a
484%  Increase 1n  testosterone  metabolism by 5  days  after  dosing and  a  19%
decrease  1n  plasma  testosterone,  the  latter  level  returning to  control
levels  by  the 15th day  after  dosing  1n spite of  elevated  mlcrosomal  enzyme
activity  by  532%  (Peakall,  1976).   For  female  rats  pretreated at  25  mg/kg
bw/day  for  7  days,  estrone metabolism  Increased by  155%  (Welch  et  al.,
1971).  Since  toxaphene  1s also known to bind to  mammalian cytochrome  P -450
to give a  typical  type I difference spectrum  (Kulkarnl et  al.,  1975),  these
effects may be attributed to Induction of hepatic cytochrome P-450.

    The toxlclty  of Toxicant B  to  mice Is  Increased  by  a  factor of  2-8  by
administration  of   plperonyl  butoxlde  that Inhibits  the  cytochrome  system
(Saleh  et  al., 1977;  Turner  et al.,  1977),  which Indicated  to the authors
that cytochrome  P-450-med1ated  detoxification mechanisms were  Important  to
explain the mode of action  of  toxaphene  toxldty (see Chapter VII).   The rat
liver mlcrosomal system  also mediates  reductive  dechloMnatlon of  Toxicant  A
and B  (Khalifa  et  al., 1976; Saleh and  Caslda,  1978).  NADPH and  relatively
anaerobic  conditions  were  required.   However,   significant nonenzymlc  reac-
tion  1n these  systems  may  also occur  as  demonstrated  by  the ability  of
reduced hematln  to cause  reductive  dechlorlnatlon (Saleh  and  Caslda,  1978,
1979).   The metabolites  of these nonenzymlc systems  have already  been  Iden-
tified  In Chapter  III, and  reductive  dechlorlnatlon  and  dehydrochlorlnatlon
occur.   Germinal and  vicinal halogenated  aliphatic organlcs are particularly
susceptible.
02030                               V-27                             02/26/87

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    Trottman  and   Desalah   (1980)   fed   male  rats   (Sprague-Dawley  175 g;
5-6/group)  diets  containing 0,  50,  100,  150 and 200  ppm toxaphene  In  the
diet  for  14 days  causing  Increased liver  weights  at  the 200 ppm  dose  and
decreased thymus weight  at  the 150 and  200  ppm dose.   In addition,  enhance-
ment occurred for  liver  hydroxylatlons of pentobarbltal  1n a  dose dependent
manner, and  for aniline and  ethylmorph1ne-N-demethylase activity In  a  non-
dose-dependent  manner   1n  the   postmltochondrlal   supernatant   fraction.
Decreased   sleeping  time   following   pentobarbltal    administration   also
occurred,  but was not dependent on dose.   Hepatic mlcrosomal  protein content
was Increased 20-35% 1n  comparison with  control  values.   In  the liver mlcro-
somal   fraction,  the activity  of  NAOPH-cytochrome c-reductase  was Increased
20-50% at doses >150  ppm.   Cytochrome P-450  levels were Increased 15-27% at
doses  >50 ppm.  There were no  effects  on the activity of NADPH-dehydrogenase
or  of  the levels  of cytochrome  b..   The  binding of  both aniline and'hexo-
barbltal  to cytochrome P-450  was  Increased by  55-210%  at all  doses.   All of
these   results show  that  specific  functions  of the  MFO  system  of  the liver,
I.e.,  cytochrome P-450 and  NAOPH  cytochrome  c-reductase  (the  sites  that  are
sensitive to  aniline  and hexobarbltal  Interactions),  are affected  by toxa-
phene.

    A  toxaphene formulation was  subfractlonated on  a silica  gel  column  Into
64  fractions  of varying polarity  (Pollock  et  al.,  1983).  The  most highly
toxic   components  to Sprague-Oawley  rats  were  found 1n the  nonpolar  frac-
tions.  Young  Sprague-Dawley  rats  (24  days;  70+6  g) were administered  two
fractions,  one  constituting 73%  (the  nonpolar  fraction)  and  the other  27%
(the polar  fraction)  at  25 mg/kg  bw as well as  unfractlonated toxaphene at
doses   of  0,  5,  25 and 100  mg/kg bw  by  1.p.  Injection  using olive  oil


02030                               V-28                             02/26/87

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carrier.  Liver weights  Increased  after  administration  of  toxaphene  doses  of
25 mg/kg  bw or more  1n_a  dose  dependent manner, as also  did  the  cytochrome
P-450   level,  amlnopyrlne   demethylase   activity   and   aldrln  epoxldatlon.
Mlcrosomal  protein  was  Increased  only  at the  100 mg/kg  dose.  The  protein
content  In  the postmltpchondMal  supernatant was  not affected  by  toxaphene.
Both  polar   and  nonpolar  fractions   caused  the  same   results  as  toxaphene
alone.   The "no  significant  Inductive  effect"  level  for  1.p.  administered
toxaphene Is between the 5 and 25 mg/kg dosages.

    Other  enzyme  systems   are  affected by  toxaphene.   One  major  system
Involves  the metabolism of sugar  to  produce high energy  compounds.   At  330
vM,  toxaphene  Inhibited the  in vitro  activity of mltochondrlal  sucdnoxl-
dase  In  beef heart  by 76% and  the  NADH-ox1dase  system  by 96%  (Pardlnl  et
al.,  1971).   At  100 yM,  the  active transport  of glucose  through  Isolated
mice  Intestine was  Inhibited  (Guthrle et al.,  1974).   Doses  of  1.2  mg toxa-
phene/kg bw/day for  6  months or an acute 120 mg toxaphene/kg bw dose to rats
altered   lactate   dehydrogenase   (LOH)   spectra   1n  the   liver   and   blood
(Kuz'mlnskaya  and  Alekhlna, 1976).   In  the liver,  the acute dose  caused  a
45% Inhibition of total  LDH activity  for 15  days postexposure.   Increases  In
the Isozymes,  LOH-1, LDH-2 and LOH-3  were  observed, LDH-4  only  transiently
decreasing and LOH-5 not at all.   In  the blood, the acute dose affected only
LDH-1.  Chronic  exposures  decreased  total  LOH  activity  1n the blood  (30%)
and  liver  (45%).   In  the   liver,  LOH-3  Increased after  3 months and  LOH-5
decreased after  6  months.   In the  blood,   LDH-1  and  LDH-2  activities  were
depressed and LDH-5  activity elevated after  6 months.   Depressed  LDH activi-
ties,  were  noted  1n  the  liver  (17%),  kidneys (18%)  and serum  (20%) of  rats
given  35  mg toxaphene/kg bw/day for  6 months  (Gertlg  and Nowaczyk,  1975).


02030                               V-29                             02/14/85

-------
 In  the  kidney,  there was a 17X  decrease  1n  LDH-1.   Decreases  of  50% In both
 serum  alkaline  phosphata_se and  liver  glutamate dehydrogenase  also  occurred
 after the 6-month exposure.

    Peakall  (1979),  who  gave  male rats an acute  120 mg toxaphene/kg bw dose
 by  capsule,  showed  that levels  of  pyruvate  and lactic add  In blood plasma
 at  1, 5 and  15  days  were not  affected.  This was so even when rats given 1.2
 mg  toxaphene/kg  bw/day  were observed  at  1,  3 and 6  months.   A steady-state
 level of  toxaphene  residues 1n  the  liver occurred after 1 month  of chronic
 dosage;  In  the  brain,  this took  1-3  months.  The  author  Implied  that  the
 rats had  to be  stressed  1n some way  to  obtain the  results  of Kuz'mlnskaya
 and Alekhlna  (1976)  and Gertlg  and  Nowaczyk (1975).   Peakall  himself,  how-
 ever, gave no data on strain of rat,  weight,  gender or age.

    Technical grade  toxaphene was  used  1n  two studies  by Srebocan  et  al.
 (1978,   1980a)   on  the  enzymatic  regulation of  carbohydrate  metabolism  In
poultry.  In  the 1978  study,  groups  of  1-day-old N1ck  chick  cockerels  (10
 birds)  were  fed  for  2 weeks on  a  formulated diet  containing  0, 0.1, 0.5,  1,
 5,  10,  50 or 100 ppm  toxaphene  (using sunflower oil  to  dissolve  the pesti-
cide and  to mix H  with  the  diet).    Inhibition of  enzymes of  the gluconeo-
genlc pathway 1n the  liver  was observed  at  1 ppm.   Enzymes  Involved 1n pyru-
vate metabolism  were  Inhibited  between 20 and  50%  (pyruvate  carboxylase,  EC
6.4.1.1J  and phosphoenolpyruvate  carboxyklnase,  EC  4.1.1.32).   There  was
not, however, a  dose response.   Fructose-l,6-d1phosphatase  (EC 3.1.3.11) and
 glucose-6-phosphatase  (EC  3.1.3.9) activities,  and  glucose  levels  were not
 significantly affected  at  p<0.05.   Pyruvate levels  were Increased  by toxa-
 phene  1ngest1on.  Based  on studies  with DDT,  the  authors   suggested  that


 02030                               V-30                             02/14/85

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 the  effects  of  a  number  of  chlorinated  pesticides  were  adrenocortkal ly
mediated.   In  the  Srebocan  et   al.  (1980a)  study,  the  same  experimental
protocol was  utilized except that  toxaphene  was  given at  5 ppm  1n  the  diet
separately  or  1n combination with  heat or  starvation stress  together  with
the  appropriate  controls.  While toxaphene  alone caused an  accumulation  of
pyruvate  (40%),  heat  stress  alone  also caused  an accumulation  of  pyruvate
(60%)  and  Increased  the  activities of  fructose-1,6-d1phosphatase  (15%)  and
glucose-6-phosphatase  (17%);  the  combined  heat  and  toxaphene  treatment
caused accumulation of pyruvate  (17%) and  Increased  the  activity  of  glucose-
6-phosphatase  (17%).   The  activities  of pyruvate carboxylase,  phosphpenol-
pyruvate  carboxyklnase,   and  the  level of  glucose were  not  significantly
affected by  any  of  the treatments.   For starvation  stress/toxaphene experi-
ments, all  the  toxaphene  treatments (5 and 10  ppm diet)  lowered  the activi-
ties  of  all  the above enzymes  and Increased  the accumulation  of  pyruvate
(70-80% excess).  However control results were  not consistent  with those for
the 5 ppm dose already quoted above.   Starvation  stress  caused accumulations
of  pyruvate  (20%)  and  enhanced   fructose-1,6-dlphosphatase  activity (100%).
The  combination  experiments  led  to  Inhibition  of   glucose-6-phosphatase
(35-40%) and  glucose depletion   (35-40%)  for  the 5  and 10  ppm  doses,  and
Inhibition  of  pyruvate  carboxylase (40%)  and phosphoenolpyruvate  carboxy-
klnase  (60%).   Fructose-1,6-dlphosphatase  levels   1n the toxaphene and  toxa-
phene/starvatlon  experiments were  not  different from  controls, whereas  a
large elevation  occurred  from starvation stress  alone.  These results  Imply
that  stringent quality assurance of the environment and diets must  be  main-
tained during toxlcologlcal studies.
02030                               V-31                             02/14/85

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    In  an  abstract,  Desalah et al.  (1979)  reported  that  male Sprague-Oawley
rats  fed 0,  25,  50  and" 75  ppm  toxaphene  In  the  diet  for  8  weeks  showed
marked  dose-dependent  Increases  1n  liver  homogenate  glucose-6-phosphatase
and fructose-l,6-d1phosphatase  In the liver mlcrosomal  fraction,  both being
maximally  stimulated  by 70%.   It  was  concluded that toxaphene  affected  the
process of gluconeogenesls.

    Alekhlna  and  Kuz'mlnskaya (1980) determined LDH activity and  changes  In
LDH Isoenzymes  In the  liver  and myocardium of  "white"  male rats (200-250 g),
dosed  perorally with  2.7  mg toxaphene/kg  bw/day,  percutaneously  at  9.4  mg
toxaphene/kg  bw,  and  a  combination  group,  all   treatments  being  over  4
months.  LDH  activity  In  myocardium after  4 months  Increased  11-34% regard-
less of  the mode  of  application,  though  the combination group gave the high-
est  Increase  compared  with  the  single  treatments  (16-18%).   LOH  activity
Increased  (32%) 1n  the liver only for the combination  treatment.   When pro-
vided  orally,  toxaphene caused  Increases  In LDH-1  (65%) and a decrease  In
LOH-5  (27%);  1n  the  myocardium,   decreases occurred In  LOH-3   (35%)  and  In
LDH-4  (70%).   When  provided  percutaneously,  Increases  1n  liver LOH-3 (64%)
and LDH-4  (81%) were  found  as well  as  In myocardium (18 and 62%, respective-
ly).  LDH-5 was decreased  (28%) only 1n  the  liver.   For  the  combined treat-
ment,  the LOH-5 content 1n  the  liver remained  decreased (25%),  but LOH-3 and
LDH-4 were  Increased  by 24  and 140%,  respectively.  Although  these changes
In myocardium were  postulated by  the authors to  be  due to toxaphene residue
deposition, this was not demonstrated directly.
02030                               V-32            '                 02/14/85

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    Male albino  Sprague-Dawley  rats  (300-350  g,  8/group)  treated  l.p.  with a
single  40  mg  toxaphene/kg  bw/day  1n vegetable  oil  carrier (0.5  ml)  showed
decreased  (19%)   plasma   cholesterol  only  60  days   posttreatment,  with  no
effect  on  plasma  trlglycerldes,  normalized  liver  weights, mlcrosomal  pro-
tein, or cytochrome P-450 (Ishlkawa et al., 1978).

    In  another series  of experiments, Kuz'mlnskaya et  al.  (1980)  determined
the oral and dermal toxlclty of  toxaphene 1n male rats  separately  and  then
Investigated concomitant  application  of  oral  and dermal  doses.   Endpolnts  of
toxlclty Included  the  activity  of  marker enzymes of  mitochondria,  mlcrosomes
and lysosomes  1n  fractions  of liver  homogenates.  Toxaphene applied In  vivo
Increased  1n  24  hours  the  activity  of   the liver mltochondrlal  enzymes  suc-
clnlc dehydrogenase  175% and cytochrome  oxldase 1000%.   Mlcrosomal  glucose-
6-phosphatase  activity  was  Increased   41%  and  lysosomal  add  phosphatase
activity  was  Increased   46%.   The   dose  (or   doses)  that  produced   these
responses  was  not  clearly   Identified  nor was  the  vehicle  reported.   The
doses (mg/kg bw)  toxaphene  used In this  study  were  as  follows:   oral,  67.5,
38.5 and 6.75;  dermal, 235,  134  and 23.5.  The greatest  Increase 1n  enzyme
activity was seen 24 hours after application.

    Hehendale  (1978)  reported  a  decrease  of  biliary  flow  and  decreased
l4C-1m1pram1ne excretion  from  the  liver  of  male Sprague-Dawley  rats  dosed
at  100  ppm toxaphene  In  the diet for 8 days.   Trottman and  Desalah  (1979)
studied  ATPase activities  1n the  brain,  kidney  and  liver  of male  ICR  mice
(30 g each)  in vivo and  \n vitro.   For  the  in vivo studies,  toxaphene  was
provided at  0, 10,  25  and  50 mg/kg  bw/day over 3 days by  corn  oil  (0.3  ml)
for 4-6  mice  1n  each group.  Enzyme  activities  were  measured  36 hours  after


02030                               V-33                             02/14/85

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the  last  dosage.  For  Jm vitro  studies,  toxaphene at  7.5,  15.0 and  30  WM
Inhibited  brain  (52-56%)  and  kidney  (42-46%)  Naf/Kf-ATPase  activities,
and   the   ol1gomydn-sens1t1ve  Mg2f-act1v1ty   In   brain   (53-61%),   kidney
(37-65%)  and  liver   (20-62%).   The  Inhibition of  Ol1gomyc1n-sens1t1ve  Mg2t-
ATPase  activity was  dose-dependent  1n  the  kidney and  liver  preparations.
The   ol1gomydn-1nsens1t1ve  Mg  f-ATPase   activity  was   also  Inhibited  In
brain  (29-45%), kidney  (47-50%) and  liver  (24-51%),  the  Inhibition  being
almost  dose-dependent  for  brain and  liver.   For  In  vivo  studies,  Na*/Kf-
and  ol1gomyc1n-sens1t1ve  and  Insensitive  Mg *-ATPase  activities  In  toxa-
phene-treated  mouse  brain  were  not  altered  significantly.  Kidney  ATPases
were  significantly  decreased   (down  to  42%  for  Na*"/Kf-ATPase  activity;
35%   for  ol1gomydn-1nsens1t1ve  Mg *-ATPase)  In  a  dose-dependent   manner
except  for  ol1gomyc1n-sens1t1ve  Mg  -ATPase.   A  dose-dependent  decrease  In
the  ollgomycln-sensUlve Mg *-ATPase  activity  (down  to  35%)  was found  In
the   liver,  but,  1n contrast   with  the  kidney, kall   doses   Increased  the
ol1gomyc1n-1nsens1t1ve component  (over 60%).   The differences between the  In
vivo  and  ^n  vitro results  for  the  brain  may arise  from  the  fact that  toxa-
phene may  be metabolized  extensively 1n  in  vivo  studies  before  1t  reaches
the  liver (perhaps  by  reduced  hematln   or  another nonenzymatlc  process),
kidney  or brain.   Rapid  metabolism may  also  account  for  the effects  on
ATPases 1n kidney  and liver.   The  crucial presence or  absence of  toxaphene
after addition  In the \t± vitro or ^n vivo studies was  not directly measured.
The  Naf/K*-ATPase  Is  postulated to  be  Involved  In  active 1on  transport;
the  ol1gomydn-sens1t1ve  Mg *-ATPase  In  mitochondria   Is  believed  to  be
Involved  1n  oxldatlve phosphorylatlon.   The Interference  of  toxaphene  with
the  production  of  ATP  may cause  an  Impairment  of transport processes In the
liver and kidney, and may be related to Impairment of biliary excretion.
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    In  another  study  (Trottman  and  Oesalah,  1983),  adult male Sprague-Dawley
rats  (175-200 g;  6/grouji)  were  fed 0,  50, 100,  150 and 200 ppm toxaphene In
powdered   chow   for   8  weeks.    Toxaphene  Inhibited   NaVKf-  and   Mg2f-
ATPases  of  synaptosomes  in  vitro  In  a  dose-dependent  manner.   The  IC.Q
values  were  about  30  yM  toxaphene  for   Naf/Kf-ATPase   and  15  yM  toxa-
                 M
phene  for  the  Mg *-ATPase.   Fifty  percent  Inhibition  of  ouabaln  and  dopa-
mlne  binding occurred  around  150 and  200  yM toxaphene,  respectively.   For
^n  vivo studies,  no  dose  responses were  observed  for  decrease  In  enzyme
activities  (30-40X decreases  for  doses  >100  ppm) even  though  all  ATPase
activities were decreased.   In fact, there  were no  significant  changes  for
ID.  v^vo  studies  Involving  ouabaln  and  dopamlne  binding  to  synaptosomes.
Toxaphene was postulated  by the authors to  have metabolized rapidly  J_n vivo
and not  reached the brain  to  explain  the difference  In the in  v 1 vo  and in
vitro  results.   Naf/K*-ATPase  has  been  shown   to  be  a receptor  of  ouabaln
and perhaps  dopamlne.   Toxaphene  Itself Interferes  in vitro but not in vivo,
and caution  therefore  must be exercised  1n  the  use  of  in vitro  tests  1n
place  of  in  vivo tests.   Fattah  and Crowder (1980)  also  tested  the  effects
of  toxaphene in vivo and in  vitro  on  the plasma membrane  ATPase  of  kidney,
brain  and  liver from  mice.  Male  Swiss Webster mice  (35  per  treatment  and
control  groups) were  administered  by  oral   gavage an  LD5Q (112  mg/kg  bw)
dose of  toxaphene  1n  corn oil, or  corn oil  alone.   Kidney ATPases were more
sensitive  to toxaphene  than   the  liver  and  brain enzymes.   The  Na*/K*-
ATPase  was  Inhibited  significantly only  1n  the  kidney,  and the Mg f-ATPase
was Inhibited significantly 1n  all  three tissues.   Twenty  mice were used to
study  the  effects of  hi  vitro toxaphene  exposure  of  tissues.   Each  tissue
was  tested  with  four  concentrations  of  toxaphene,  10~«,  10~5, 10"6  and
02030                               V-35                             02/14/85

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TO"7 M,  and  two  controls.    Three  samples  for  each treatment  were run  In
each  of  the  three  tissues.   As observed  following  in  vivo  exposure,  toxa-
phene  significantly   Inhibited  the   Naf/K*-ATPase   activity  only   In   the
kidney  homogenates,  In   this  case  with  a  dose  effect.   Mg2*-ATPase  was
significantly reduced  1n  all tissues;  In  the Hver   homogenates  only  10"4  M
toxaphene significantly Inhibited activity.

    Parvu et  al.  (1980)  reported In an abstract that toxaphene given  orally
to white Ulstar rats at 4 mg/kg  bw/day for 30 days caused I1p1d accumulation
In  the  liver,  depletion  of free  fatty  adds,   and an  Increase  of "liver
cholesterol.

    Toxaphene may also affect VHamln  A  storage.   A  concentration of  100  ppm
toxaphene  In  the  diet  for 72 days  to 28-day female  weanling rats  (Phillips
and Hatlna,  1972)  was  followed  by  mating  with untreated males.  At  20 days
postpartum,  liver  Vitamin A  levels  relative to controls were  depressed  by
11% (p<0.01) In newborn rats, but not 1n dams.

    Alterations In clinical  chemistry  have  also been  seen In  subchronlc oral
toxaphene exposures.   Mice  with no clinical  signs of Intoxication  evidenced
consistent  Increases   1n  serum  add  phosphatase, glutamlc  pyruvlc  trans-
amlnase  and gamma-glutamyl   transpeptldase  activities,  along with  Increased
neutrophll  counts  and  changes 1n urine  protein  (Blumler, 1975).  At  a much
lower dally dose, rats had only  a  transient Increase  1n serum alkaline phos-
phatase during the  fifth  month  of  1ngest1on and  showed  no variation  In urine
hlppurlc  add (Grebenyuk,   1970).    Increases 1n  all  of the  above  enzyme
activities  are  consistent  with  the  mild   liver  pathology   associated with
subchronlc  toxaphene exposure.
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 Teratoqenldtv and Reproductive Effects
    Hamma1s.   The  multtgeneratlon  reproductive  effects  of  toxaphene  and
 two  other  pesticides were  Investigated  by  Kennedy et al.  (1973).   two con-
 trol groups and  two  test  groups  of Sprague-Dawley albino rats consisted of 8
 male and 16 female animals  each.   The treated groups  started receiving 25 or
 100  ppm toxaphene 1n  the diet  (adjusted  to contain  2X corn oil)  added to
 commercial  chow  at  28  days  of  age.   FQ  parental animals  were exposed  to
 toxaphene  for  42  weeks  and  then  sacrificed.    F,  and  F.  parents  were
 exposed  for  39  weeks and  then  sacrificed.   Animals were first  mated  at  100
 days of age.   First  litters  were  reduced  to  10/Htter  on postpartum  day 5
 and  retained  for 21  days  until  weaning.  After a  10-day rest,  the parental
 animals were mated again  to obtain second  Utters.   At weaning of the second
 Utters, 8  males and  16  females  were  randomly  selected from each  group as
 parental  animals for  the  next   generation.   This  procedure was  continued
 through  three  successive 2-l1tter  generations.   Toxaphene  had  no  effect on
 Utter  size, pup survival or  weanling body  weights.  Parental  animals at  100
 ppm  showed  liver changes consisting of slight  cytoplasmlc  fatty vacuollza-
 tlon In 63% of animals examined.   This  change was not observed at 25 ppm and
was not  accompanied  by adverse  effects  on  growth, mortality, organ weights
 (liver, kidney,  spleen, gonads,  heart,  brain,  adrenal  glands,  thyroid gland)
or  among  the corresponding   organ-to-body  weight  ratios  or  reproductive
capacity.   No  differences  were  observed among  test  and  control  groups  of
F- parental  animals  with  regard  to  total,  free and  esterlfled cholesterol
concentrations,  hematologlc  parameters,  urine  analysis  and  other  clinical
chemical  findings.    Tumor  Incidence was  not  affected and  there  was  no
evidence  of  teratogenldty   among  test progeny.   The  no-effect  level  was
judged  to  be  25  ppm 1n the diet for toxaphene  (1.25  mg/kg  by assuming food
 Intake equal to 5X of body weight).

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    Chernoff and  Carver  (1976)  used rather high doses  of  toxaphene  to study
 fetal  tox1c1ty  of  the pesticide 1n rats.   CO rats were  administered  toxa-
 phene  1n  corn oil  (0.1  ml)  by  gastric Intubation  on  days  7 through  16  of
 gestation at doses  of 15,  25 or  35  mg/kg  bw.   The number of  rats  per  group
 were  as  follows:   controls,  33  rats;  15  and  25  mg/kg,  39  rats/group;  35
 mg/kg,  16  rats.    At 35  mg  toxaphene/kg,  toxldty  was  evidenced  by  31X
 maternal mortality.   There  was  also a  dose-related  reduction  In  weight gain
 of dams (p<0.001) at  15  and  25  mg/kg/day  and  fetal weight  (p<0.001)  at 25  mg
 toxaphene/kg/day.   Even  though  there  was  significant  maternal   toxldty  In
 all treated groups, there were  no dose  related changes  1n  fetal  mortality  or
 1n the occurrence of anomalies.

    The  rat fetus  was  shown  to contain  a  small  amount of  14C-toxaphene
when  assayed  5  days  after  an  oral  dose  of 2.6  or 6.5 mg/kg maternal  body
weight onv day 15  of gestation (Pollock  and HUlstrand,  1982).  Fetal  tissues
contained 28  yg/kg wet  weight;   In  comparison,  maternal   fat  contained  7.48
mg/kg tissue.

    Mixed-strain  white  rats  and golden  hamsters  were used  by   Martson  and
Shepel'skaya (1980b)  to  evaluate the embryotox1c1ty of  toxaphene.  Rats were
given 4  mg  toxaphene/kg bw/day by  gastric  Intubation  during the period  of
organogenesls (days 6-15) and during the  entire pregnancy  (1-31  days)  and  to
hamsters on days 7-11 and  1-15  of  gestation.   In rats,   no  adverse  effects
were  noted  on  development,  fetal  weight,  ratio of males  to  females,  or  the
number of fetal  deaths.   In hamsters,  the  only toxaphene-related effect  was
reported as an Increase In an unidentified developmental anomaly.
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    Badaeva  (1979,  1981}  examined  the effects  of toxaphene on chollnesterase
activity  1n  cardiac neural  structures  of  dams and  future  offspring  on days
14 and  20  of  gestation.   A group of 12 pregnant animals was perorally admin-
istered dally  doses  of 12 mg toxaphene/kg  bw.   It  was  found that the number
of dead embryos was  higher when  toxaphene  was  administered throughout gesta-
tion  than  when administered  between the 6th and 14th day of gestation.  When
given  throughout  gestation, the  amount of  toxaphene  found In  the  maternal
heart  was  higher   on  the  20th  day  of  gestation  (14  yg/g),   uterus  (9.7
yg/g),  brain  and  spinal  cord   (4.3  yg/g)  than   on   the  14th  day.   The
number  of  pups per  Utter  (6-8)  was less  1n  treated groups  than 1n  control
groups  (10-11);  birth  weight of  pups  was  depressed and  Increased mortality
was noted  1n  the  first week postpartum.   In treated embryos,  chollnesterase
activity along the myocardlal vessels  and  cells of  the  atrloventrlcular node
was almost absent  at 14  days of gestation;  at  20 days,  there  was a  delay In
differentiation of  fetal  cardiac neural elements and a  continued depression
of chollnesterase  activity.  On  the  20th  day, 50-60%  of  the cells  of  the
cardiac ganglion of  pregnant treated animals exhibited  decreased cytoplasmlc
chollnesterase activity.

    Toxaphene was  one  of   five compounds administered to pregnant 90-day-old
CD rats during the  period  of organogenesls  (7-16 days  of gestation),  and the
effects on organ differentiation were determined In day  21  fetuses  (Kavlock
et al., 1982).   Two dose   levels  of  toxaphene  1n corn oil,  12.5  and  25 mg/kg
bw/day, administered perorally to a group of  five  animals,  had  no effect on
the average  number  of Implants,  fetal mortality  or fetal  weight.   No soft
tissue  abnormalities  were,  observed.   There was  no statistical  difference
between  the  following parameters   1n  treated  and  untreated rats:   brain


02030                                V-39                             02/14/85

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weight,  total   DNA  and  total  protein;  lung weight  and  primary  surfactant
materials;  liver  weight "and total glycogen; and  kidney  weight.   There was a
statistically  significant  decrease  1n  alkaline phosphatase  activity  of  the
kidney  at  25 mg  toxaphene/kg bw  and 1n total  protein 1n  the kidney  at both
doses.

    Behavioral  effects of  low levels of toxaphene,  and Us toxic components,
Toxicants A  and B,  were studied  1n  juvenile albino rats exposed perlnatally
(Olson et al.,  1980).  Toxaphene,  or Toxicant  A or  B, was given dally 1n the
feed,  starting  on day 5 of gestation.  Pups were  exposed to dietary levels
after weaning  until  the  study was terminated at  3  months postpartum:  toxa-
phene  50 yg/kg  bw,  Toxicants  A  and  B,  2.0 yg/kg bw.    Because  sufficient
supplies of  Toxicant A could  not  be  procured.  Toxicant B was substituted for
Toxicant A  when supplies were  depleted (rats  were  40 days  old).   Offspring
from  12  mothers made up  three  treatment groups  of 16 each  and  one  control
group  of  15.  Behavioral  testing  began when the offspring  were  7  days old.
Two  different  behavioral  test  periods were studied:  early  development with
pups  being   tested  on postnatal  days  7  through 17  (swimming  and  righting
reflex); motivational,  learning  and retention  tests  on  days 70  through  90
(symmetrical  maze).    In  their   early  development,  treated animals  showed
retarded maturation  based  on their  performance  In  overall  swimming  ability
1n comparison  with  control animal performance  (p<0.0001  on  day  10).   By day
16 all  groups  displayed normal  swimming ability.   Inferior  swimming  ability
may reflect  an  ability of  toxaphene  compounds  to affect  early functional and
behavioral  development  of  the  CNS  In   the   Immature   rat.   Toxaphene-fed
animals  constituted  the only  group  to  exhibit  overall  retarded  righting
ability  1n  tests conducted from  days  7 through 17.   On  day  15  the  order  of


02030                                V-40                             02/14/85

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decreasing superiority was  as  follows:   controls,  Toxicant  A,  Toxicant 8 and
toxaphene (p<0.0001) with each  group  being  significantly superior  to the one
following.   Inability  of the  Immature  rat  to metabolize the  simple mixture
may account  for  the  Increased  toxlclty  of toxaphene.   The test of motivation
revealed no  significant  differences between any of the  groups.   In the maze
retention test.  Toxicant A  animals had  no  difficulty  1n  learning  the test
problems but. were  Inferior  to the  other  groups  In retaining that knowledge.
Behavioral changes were  observed  at dosages  far  lower than  those used  In any
previously reported study.

    In another  behavioral  study,  Crowder et al.  (1980)  also reported a sig-
nificant  difference  In  the  length of  time for  treated animals  to become
positive In  the  righting reflex.    In the Crowder  et  al. (1980)  study, three
pregnant rats  received  dally  oral  doses of 6 mg/kg  bw 1n 0.1 mi  corn oil
from  day 7  of  gestation  until  parturition.   Testing  methods  Included  a
simple two-choice maze,  motor  skills, an open field test and other behavior-
al tests.

    Investigators  In  Russia  found  no  effect  on  behavioral   responses  of
1-month-old  pups  exposed   during   days  6-15  of  gestation  (Hartson  and
Shepel 'skaya,  1980a).    The  dose  was  reported  as  the 0.1  L05Q  given  by
gastric  Intubation  to  six  pregnant  animals;  however,  the LD50   for  the
mixed strain  rats  was not  given.  In this  same  study,   6  male  and 6  female
rats  received  4  mg toxaphene/kg  bw by  gastric  Intubation dally for 10 weeks
before  mating.   Hales  and  females  receiving   toxaphene  were  bred  with
untreated  rats.   At  the  age  of  2  months,  offspring  were monitored  for
behavioral response.   There were  no  toxic effects  observed In the offspring


02030                               V-41                             02/14/85

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as  judged  by  locomotion,  grooming and rearing  activities.   The  authors  con-
cluded  that  toxaphene did  not have  major  gonadotoxlc effects.   There  was,
however,  an  Increase  In  the  number   of   sperm  with  abnormal  morphology
(Martson and ShepeVskaya, 1980b).

    OlPasquale  (1977)  examined the effects  of  toxaphene on  fetal  guinea pig
development.   In  this study,  toxaphene  was administered orally  to pregnant
females  at  a dose  of  15  mg/kg bw from  day 21  to  day 35 of  gestation.   No
effects  were  noted on  the  anatomical development  of the fetus.   There was
some  fetotoxldty  as  shown  by  adverse effects  on collagen-related structures
of  the  fetus.   This was  attributed  to a functional  deficiency  of vitamin C
related  to MFO  Induction.  Maternal  guinea  pigs showed a slight loss of  body
weight,  but  no effects  attributable to   toxaphene  exposure  were seen  on
maternal liver weight or mortality.

    CD-I  albino mice  were  administered  toxaphene  In corn  oil  by  gastric
Intubation during  the period  of  organogenesls, days 7  through  16 of  gesta-
tion  (Chernoff  and Carver,  1976).  The  number  of  mice per  group and  dally
doses of toxaphene  were as  follows:   75  mice,  0 toxaphene;  26 mice, 15 mg/kg
bw; 45  mice,  25 mg/kg bw; 90 mice,  35 mg/kg bw.  Mice were  killed on  day 18
of  gestation  and  fetuses weighed and prepared for  soft tissue  or skeletal
analysis.  In the  35 mg/kg  group  there was  8% maternal mortality.  There was
a dose-related  (p<0.01)  reduction 1n the average maternal weight  gain  and a
dose-related  Increase  1n  the  I1ver-to-body  weight  ratio.   There were  no
significant dose-related  responses  In fetal mortality,  fetal weight,  number
of  caudal  or  sternal ossification  centers,  or  Incidence  of supernumerary
02030                               V-42                             02/14/85

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 ribs.  Five Utters  from  dams  receiving the  high  toxaphene  close had  one  or
 more  fetuses  with encephaloceles;  none were observed  1n controls or  other
 treated groups.

    KepHnger et  al.  (1970)  conducted a 5-6  generation  study  of five pesti-
 cides  Including  toxaphene and  combinations  of  pesticides using  Swiss  white
 mice.   Offspring  were  exposed  in utero.  by  the mother's milk, and  1n  the
 diet after pups were  weaned.  Toxaphene (25  ppm 1n  the diet) was fed  to mice
 during  the  entire study Including mating, gestation  and  lactation.   Animals
 not  used  for subsequent  breeding were  sacrlfled  for chemical  analyses  and
 histology.  Eighteen  120-day-old  mice,  4 males  and 14  females,  made  up each
 group.  For each  successive generation,  parental groups  were remated  to pro-
 duce  second  Utters  and the next parental groups were  selected from second
 Utters when 120  days  of age.   Little or no  adverse effect was noted  through
 five generations  of mice fed 25  ppm  toxaphene In the  diet Including the fol-
 lowing  parameters  for each generation:   Utter size, embryonic  death,  sur-
vival and  body  weight.  There were  no  statistically  significant differences
between several Indices calculated by the  Investigators  Including viability,
 lactation, survival  and reproduction.   H1stolog1cally there was  evidence  of
fatty  changes   In  the  liver,  especially  In the  central zone.  There  was
 little or no damage to the brain.

    Twenty-eight compounds of known  teratogenlc  potential  were assayed by  an
In vivo  teratology screening  procedure  (Chernoff and  Kavlock,  1982).   Toxa-
phene In corn oil  was  administered by oral gavage  (75 mg/kg  bw) to 25 gravid
CD-I   mice  on days 8-12 gestation,  the period  of  organogenesls.  Two  mice
died  before  term  and  only  11   were pregnant.    Dams were  allowed  to  give


 02030                               V-43                             02/14/85

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birth, and  Utter  size and weight  on  postpartum days  1 and 3 were  compared
with  concurrent  controls.   Results  Indicated significantly  reduced  maternal
weight change  and  reduced fetal weights  on  postpartum day 1.   There was  no
difference  1n  the  body weights  of  treated versus control  pups on  postpartum
day 3.

    Allen et al.  (1983)  studied the effect of  toxaphene on  Immune  responses
of  offspring   of  mice exposed  orally 3  weeks   before  breeding,  and during.
gestation and  lactation.  Toxaphene  In  acetone  was  mixed  Into  ground  rat
pellets  to  attain  a  concentration  of 10, 100  or 200  ppm.  Transplacental/
lactation exposure to  toxaphene produced  some degree of Immunosuppresslon  In
the pups, even though overt  toxlclty of the  Insecticide was not apparent  at
the lower dose.  At  10 ppm there was  a  significant depression of  macrophage
phagocytosis (p<0.05)  but  no  effect on hypersensUlvHy responses  and  anti-
body  tlters  to bovine serum  albumin.   Relative  degree  of  Immunosuppresslon
was  greatest   In   macrophages  followed  by  humoral  Immunity;  cell-mediated
Immunity was least affected.

    Birds.  The effects  of  toxaphene have  been examined  1n a  number  of
bird  species.   White  leghorn  female chicks, 1  day  of  age,  were   fed  diets
containing  0,  0.5, 5, 50  and  100  ppm toxaphene  (Bush  et  al., 1977).   Each
treatment group  contained 90  randomly  selected  hens   that  were  bred  at  23
weeks  of  age.   Even  at  100 ppm, toxaphene  did not  significantly  alter  egg
production,  natchabllHy  or  fertility.   Arscott et  al.  (1976)  added  0,  10
and 100  ppm toxaphene to the feed  of  white  leghorn layers  (48/group) for  24
weeks.   Except for a  slight decrease  1n egg production,  no adverse effects
were observed  on fertility, hatchabllHy and  survival  of progeny.


02030                               V-44                             02/26/87

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    Three  groups  of  15 pairs of  1-year-old  black  ducks  (Anas rubrlpes) were
 fed ad  libitum w1h commercial duck  breeder  mash coated  with  0,  10 or 50 ppm
 toxaphene  beginning 90  days  prior to laying  (Mehrle et al, 1979).  Toxaphene
 was  dissolved  In propylene glycol  equivalent  to IX  of  the   total  diet.
 Reproductive  success   of   all  birds  was  followed  throughout   the  breeding
 season.  Toxaphene did not  affect reproduction or  survival.   Ducklings were
 fed starter  mash  containing  the  same concentration  of  toxaphene as  the diet
 the  parents   received.    Duckling   growth,   as  Indicated  by   weight,  was
 decreased  significantly 5 and  14 days  after hatching  In the   group  fed  50
 ppm, and was  Increased  significantly at  28 and 42  days after hatching 1n the
 group fed  a  diet  containing 10  ppm toxaphene  In  the  diet.   Duckling weight
 was not  altered  by toxaphene In measurements  made more  than 42 days  after
 hatching.   Backbone  development  was  Impaired by  toxaphene  within  14  days
 after  hatching and  collagen  was decreased  significantly  1n   the  cervical
 vertebrae  of ducklings  fed 50  ppm.   In  contrast to the effects  on vertebrae,
 toxaphene exposure did not alter  tibia development.

    Forty-five  pairs  of 1-year-old  black  ducks (Anas rubrlpes)  were placed
 on a diet  of duck breeder mash  containing 0, 10 or 50  ppm  toxaphene (Heinz
and  Flnley,   1978).   Toxaphene   was  dissolved  1n  propylene glycol  before
 Incorporation  Into the  feed  at 1 part/99  parts feed.   About  3  months later,
hens began to  lay eggs and  the  eggs were  Incubated.  Ducklings  were brooded
by hens  and  ate  a diet of duck  starter mash  containing  the  same concentra-
 tions of toxaphene as  those  fed  the  hens.   At 5 days  of age, ducklings were
 tested   for avoidance  behavior.   There  were  no  significant differences  1n
avoidance  behavior between control ducklings and  those treated  with  10 or 50
 ppm toxaphene.

 02030                               V-45                             02/14/85

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     In  a  similar  study conducted over a  19-month  period,  which Included two
breeding  seasons,  three Tets  of  15  pairs  of black ducks (Anas rubrlpes) were
given 0,  10  or  50  ppm toxaphene  In  dry mash (Haseltlne et al., 1980).  Toxa-
phene was  dissolved In propylene glycol  before Incorporation  Into  the feed
at  IX  of  feed  by  weight.   Adult survival was  not affected,  but  the weights
of  treated males were  depressed  during the summer  months.   At the end of the
second  breeding season females Ingesting  10 or  50 ppm had larger livers than
controls.  Drakes  fed 50  ppm In the  diet exhibited  elevated  brain weights
that were not  apparent   1n  50  ppm females.   In  both  years  of  the study,
breeding  pairs  fed  50 ppm  toxaphene  In  the  diet appeared  hyperactive but
caging  facilities   did not  allow  for  testing  of   this  possibility.   Eqg
production,  fertility, hatchabllHy,  eggshell  thickness, growth and survival
of  young   did  not  vary   significantly with  toxaphene  1ngest1on   In  either
breeding  season.   The  mean number of  days  required  to complete a  clutch was
lower 1n  birds  fed toxaphene than  In  controls.   Clutches  o'f  hens  fed 50 ppm
toxaphene  showed  Improved hatching success  In  the second year  of  the study
compared with' the first year.

    Toxaphene was  Injected at  levels of  0.1, 0.5, 1.0,  5.0  and 10 mg/kg egg
weight   Into  groups of 20  fertile Nick chick eggs on  day  0  of  their Incuba-
tion until day  14  (Srebocan et  al.,  1980b).  The Injection  was made 1n the
air  sac and contained 0.1 ml of the  toxaphene  solution In  sunflower oil.
The  following  assays  were carried  out  on  nine  or  more  embryo  Hvers  per
treatment:   pyruvate,  pyruvate  carboxylase,  phosphoenolpyruvate  carboxy-
klnase, fructose-1,6-d1phosphatase,  glucose-6-phosphatase and  glucose.  The
effect   on carbohydrate  metabolism  was  similar   In  all  groups.   Increased
pyruvate  concentration, decreased glucose concentration and  decreased enzyme

02030                               V-46                             02/26/87

-------
activities,  particularly  1n  the  embryos  Injected  with  the  higher  doses,
Indicated  that  toxaphene"  significantly  Inhibited the  gluconeogenlc  pathway
In  the  embryonic  liver.   Lower  reproductive  success  might   also  cause
embryonic death or a lowered vitality of hatched chicks.

    Hoffman and  Eastln  (1982)  determined the effects  of externally  treating
mallard  (Anas platyrhychos)  eggs  with   formulations  and concentrations  of
toxaphene  similar  to  those In  field applications.   Toxaphene,   In  aqueous
emulsion  applied  on  day  3 after  the  eggs were  placed  In  an  Incubator,
resulted In significant  mortality  (p<0.01)  even at one-half the  field  level
of  application   (12.5  Ib/acre  at  100 gal/acre);  the  IC™  was   108  Ib/acre
(121  kg/hectare).    At   concentrations   greater   than   the  LC5Q  there  were
abnormal survivors that  exhibited dislocation of  Joints.   Treatment  on  day 8
at  5  times the  field  level  resulted 1n embryotoxlc  effects  Including  53%
mortality,  a  reduction  In growth,  and  an  Increased  Incidence   of  abnormal
survivors (18%), some with joint defects.  When  toxaphene was applied  In an
oil vehicle either at day  3  or  day  8,  the LC^s were  greater than  for  the
concentrated  formulation  (66  Ib/A).  Treatment on  day 8  resulted  In  growth
reduction (p<0.05).  Higher concentrations  resulted  1n brain,  bill  and  Joint
defects.  Exposures  up  to 5  times the  field level  did not  grossly  affect
cervical vertebrae as revealed by alizarin red staining.

Hutaqenldty
    The  National   Toxicology  Program  (NTP,  1983)  reported  toxaphene  to  be
mutagenlc when assayed  by  the Salmonella/mlcrosomal  reverse mutation  assay.
Hill (1977),  summarizing tests  done by Litton Blonetlcs  for Hercules,  Inc.,
Indicated that  toxaphene  was directly  mutagenlc  only  for  Salmonella  typhl-
02030                               V-47                             02/14/85

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muMum  strains  TA98   (which  detects  frameshlft  mutagens)  and TA100  (non-
specific).  By  contrast,-a  "high  temperature"  toxaphene  (high boiling compo-
nents) was mutagenlc only when  rat  Hver  preparations  (S9)  were Incorporated
1n  the  assay  to  provide  for  metabolism of  the  compounds  to biologically
active forms.   Hooper  et al.  (1979)  used Salmonella mutagenlclty  assays  to
Identify  toxaphene  components  of  potential   carcinogenic  hazard.   TA100
proved to  be  the most  sensitive  Indicator  of toxaphene mutagenlclty  and  H
was  determined  that mutagenlclty  resided 1n the polar  fraction.   Mutagenlc
activity was  reduced  by 50%  upon  addition  of rat  or  mouse  S9  to  the assay
plates.   A major  toxic  component of  toxaphene. Toxicant  B,  was not mutagenlc
for  any of  five Salmonella  strains encompassing three DNA  sHes.   Treatment
of toxaphene with ethanollc  KOH 1n molar ratios of  1:1  or  1:10 for 24 hours
at 25°C  resulted  In dechlorlnatlon  of the  major  
-------
    Griffin and  H111  (1978)  described use of  an  in  vitro  DNA breakage a-ssay
utilizing  purified  covalent  closed circular ONA  molecules  of plasmld ColEl.
Breakage  rates  were determined  after  analysis by alkaline  sucrose gradient
centrlfugatlon.   Breakage  rates  significantly  greater   than  control  were
obtained  with  three mutagenlc  alkylatlng  agents  and  4/11  pesticides tested
(dexon, dlchlorvos, malathlon  and  methyl  parathlon).   Toxaphene Incubated at
0.1  mg/mt  1n  hexane with  plasmld  DNA  for  an  unspecified  amount  of  time
caused no  Increase In breaks.

    Epstein et  al.  (1972) used  a  modified dominant lethal assay  In  mice to
evaluate  the  mutagenlc  potential of  a  variety of chemical  agents  Including
toxaphene.  In  this  study,  four groups of male  ICR/Ha Swiss  mice  were given
toxaphene  either IntraperHoneally  (single doses  of  36  or  180   mg/kg)  or
orally (five  doses  of  8 or 16  mg/kg/dose).   After  dosing,  the treated males
were mated  to throe untreated  females/week  over  an 8-week  period.  Females
were dissected 13 days  following the  midweek  of their  caging with  the males.
Based on measurements of  early  fetal  deaths  per pregnancy  and the  percentage
of  females with  early   fetal  deaths,  there  was  no  significant  difference
between control  and  toxaphene  treated groups.  Thus,  1n this strain  of mice
toxaphene  apparently  does  not  produce  chromosomal  abnormalities  In  sperm
that preclude zygote development.

    Sobtl  et  al. (1983) assayed organochloMne pesticides  for  their  ability
to  cause  sister  chromatld  exchanges  (SCE)  1n a human lymphold cell  line,
LAZ-007,   of B  cell  origin.    Significant  Increases 1n  numbers of SCE  were
observed   with   toxaphene   concentrations   of   TO"5  and    10"* M.    SCE
frequency,  however,  did  not  reach  the  level  of 2 times  control  frequency


02030                '               V-49                             02/14/85

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that has  been  recommended  as the criterion  for  a  positive  response  (Latt  et
al.,  1981).   Samosh  (T974)  reported   Increased  frequency  of  chromosomal
aberrations  1n  lymphocyte  cultures  obtained from  eight  women  occupatlonally
exposed  1n  a spraying  operation  to 2  kg/ha (13.IX  1n  exposed vs.  1.6%  \n
control).    Aberrations  consisted   of   acentric  fragments   and   chromatld
exchanges.   This  1s by  contrast  to a  U.S.  EPA  (1978)  study  that  found  no
significant  differences  In  rates  of chromosomal  aberrations   In  leukocytes
between  groups  of  Individuals  occupatlonally  exposed  to  toxaphcne  and
unexposed groups.

Cardnoqenlclty
    The most definitive  studies of toxaphene carc1nogen1c1ty  were  performed
by Tracor JHco  Co.  under  contract  from the National  Cancer Institute  (NCI,
1979)  1n  spite  of  the  fact  they were  not  conducted 1n accordance  with  NCI
guidelines  (control  groups 'contained  only 10 animals  each;  pair-feeding  was
also  not  done).   In  this   study  both   genders  of  Osborne-Mendel   rats  and
B6C3F1  mice  of  age 35  days were used.   Each experimental  group contained  50
animals  of   each  gender, and   the  pesticide was  added to  the  diet as  an
acetone solution,  2% corn  oil  also  being  added as  a  dust  suppressant.   The
high and low male  rat diets  Initially contained  2560 and 1280  ppm toxaphcne,
respectively, and  the corresponding diets for females  contained  1280 and  640
ppm, respectively.   In  the  case of  mice the corresponding  high  and  low dose
figures were 320 and 160 ppm  diet  for  both sexes.  Owing  to  overt  toxlclty
these  concentrations  were  later  lowered.    In  male rats  the  high  dose  was
lowered to  1280  ppm at  2 weeks, and to  640  ppm  at  55  weeks after Initiation
of the  study.   The low  dose was  similarly  lowered  to 640  ppm  diet after  2
weeks and 320 ppm  55 weeks after  feeding had begun.   In the case of females,


02030                               V-50                             02/26/87

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both  the doses  were halved  after  55  weeks,  and  for  both  sexes  pesticide
treatment  was discontinued  after  80 weeks.   Subsequently the  animals  were
fed  control  diets without corn  oil  for 20 weeks and then  control  diets  for
an  additional 8 weeks.   In  male and female mice  both  doses were  halved  19
weeks  after  treatment was Initiated; toxaphene  treatment  discontinued after
80  weeks,  and animals were  fed  control diets without  corn oil for  7 weeks
then diets with 2% corn oil for an additional 3-4 weeks.

    Animals  that  died during  the  study,  and  also all  animals  surviving  at
the  termination  of  the  study,  were  submitted  to  pathologic  evaluation  of
major  tissues, major  organs and  all  gross  lesions.   In  the male rats, 90% of
the high  dose,  94% of the low dose and  all  of the  control  group lived until
at  least  week 52  of the study.  In females  96%  of  the  high dose,  92% of  the
low dose and  all  10  control  animals  survived beyond the 52nd week.  Although
none of  the  tumors qbserved  1n  treated  animals  were  uncommon for  the animal
strain  used,  certain tumors  and  hyperplastlc   lesions   were   present  with
higher  Incidence  In  the  treated animals.  These Included  thyroid  folUcular
cell adenomas  and  carcinomas  (7/41  low-dose, 9/35  high-dose  and 1/7  control
males;  1/43   low-dose, 7/42  high-dose  and 0/6  control  females)  and  hyper-
plaslas (3/41  low-dose, 3/35  high-dose  and 0/7 control  males; 5/43 low-dose,
3/42 high-dose and 0/6 control  females).  The  thyroid folUcular-cell hyper-
plaslas were  observed only In  the  treated  animals  and were at relatively low
Incidences.   Therefore  It could not  be concluded  that  Increased  Incidences
of neoplasms  or  prollferatlve lesions were a result  of toxaphene  treatment.
These  authors regarded  adenomas  as neoplastlc lesions;  however,   this  is
controversial.   Taking  thyroid  folllcular   cell  adenomas  and  carcinomas
02030                               V-51                             02/14/85

-------
 together,  a  statistically  significant  Increase  was found  for  the high-dose
 group  compared  with  th.fi  matched  controls  for  both  male  and  female  rats
 (Tables  V-8  and  V-9,  respectively).  In comparison with  values  from histor-
 ical  controls  from  the same  laboratory,  statistical  significance  was  also
 found  for  the  above lesions.  In  the  female  rats  there  was also a statisti-
 cally  significant  Increase  1n  the cumulative  Incidence  of  tumors of  the
 pituitary  (chromophobe  adenomas, chromophobe carcinomas  or adenomas)  In the
 high dose  compared  with the control group, although  H  1s  warranted only to
 refer  to  the carcinomas as neoplastlc  lesions.   Due  to  the high spontaneous
 Incidences  seen   In  controls,  NCI  does  not  conclude  that an  association
 exists  between   toxaphene  administration  and  an  Increased  Incidence  of
 pituitary  tumors.  From the  above  study the  authors concluded that toxaphene
 administration was associated with an Increase In thyroid tumor Incidence.

    Following  an   Independent   examination   of  h1sto1og1cal  preparations,
 Reuber (1979)  came  to  similar qualitative conclusions,  reporting 10,  15 and
 28% thyroid  adenoma and 6,  18 and  18% thyroid carcinoma  Incidence 1n control
male rats  or 1n  groups  Ingesting  the  low  or  high dose of toxaphene. respec-
 tively.   In   females,   the  corresponding  figures  were   6,  18  and  19%  for
 adenoma,   and 10, 9  and  20% for  carcinoma  Incidence.   In  addition,  Reuber
 (1979)  claimed   Increases   In   benign   and   malignant  tumor  Incidence  In
 endocrine  organs,  reproductive  system  and   mammary  glands   In  toxaphene-
 treated  rats.    Until   differences  between  Reuber's   criteria  and  those  of
 others are resolved,  however, 1t will  be  difficult to draw conclusions from
 his findings.
02030                               V-52                             02/14/85

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ro
CJ
O
                                                                                 1ABIE V-8

                                                              Incidence of lumors In Hale Rats fed loxaphene*
Dose
(ppm diet)

0 Hatched
Low
High
Number
Initial

10
SO
SO
of Animals
Examined

9
47
45
Skin Hemato-
potetlc

0/10 0/10
2/50(4X)d 0/SO(OX)
1/4S(2X) 2/45J4X)
Kidney Pituitary

0/9 0/7
1/45(2X) 0/42
1/4S(4X) 1/31 (3X)
Adrenal
HAL1GNAN1
0/9
1/4K2X)
0/37
Thyroid

0/7
1/41(2X)
2/35 (W)
Prostate

0/9
0/37
1/35 (3X)
Liver'1 Husculo-
skeletal

1/9(1 IX) 0/10
6/44 (14X) 0/50
4/45(9X) 1/45(2X)
Heso-
thelloma

0/10
0/50
0/45
Nervous
System

0/9
0/50
0^45
Totalc

0
7
10
      0 Hatched
       Low
       High
             10
             SO
0/10
0/9
                                                          0/9
             BENIGN
l/7(14X)e    2/9(22X)   l/7(14X)f
2/7(29X)9    2/9(22X)
47     2/50(4X)   3/4S(7X)  2/45(4X)   1/42J2X)*    4/41(10X)n
                                      12/42(29X)9
                                                                             0/9
0/37
                                                                                             1/26(4X)J
             SO        4S     1/4S(2X)   3/42(7X)   2/4S(4X)    4/31(13X)9   3/37(8X)n  7/3S(20X)f*k  0/3S
                                                                           1/37I3X)1
          0/9
                                                                                                                     0/44
                                                                                                                     0/4S
0/10
                                                          0/50
0/10
                                                                                         0/50
0/9
                                                                                                              2/SO(4X)   27
                                                           1/4S(2X)   1/4S(2X)  4/4S(2X)   20
 00
       'Source:  NCI.  1979
        Liver  *  neoplasttc nodules
       clotal  =  total anlMls with  tuw>rs
       tf	Muaber of anlaals  bearing  iumor  stated	
        NuHber of  anlMls with  tissue stated  examined Microscopically
        folltcular  cell  adenoma
       ^Chromophobe adenoma
        Cortical  adenoma
C-cell adenoma
Adenoma of parathyroid
Adenoma » carcinoma Incidence significant (Eisner  exact  text,  p-0.008)  compared with matched controls
Pheoc hr omoc y t oma

-------
                                                                                 TABLE V 9

                                                             Incidence  of  Tiwors  In Female Rats Fed Toxaphene*
IVJ
o
OJ
0 Niwber of Anlaals


Dose Skin Lung Liver Pituitary Adrenal Thyroid MaMury Uterus Ovary Kidney Tolalb
(ppadlet) Initial Examined

0 Hatched 10 10 0/10 0/9 1/10J10X) 0/8
Low SO 49 l/50(2X)e 0/46 4/42(10X)d 0/41
MALIGNANT
0/8 1/6(17*) 0/10 0/9 0/8 0/8 1
0/44 0/43 1/50(2*) 1/41(2*) 0/40 0/49 4
ifl
      High
0 Matched
Low
High
              SO
                    10
                    SO
                    so
49
10
49
49
                     1/42(2X)<

3/49(6X)   1/48(2X)  4/40(10X)    l/39(3X)f       2/43(5*)    0/42

                                             BENIGN

0/10       0/9       0/10         1/8(13X)9       0/8        0/6
                                  2/8(25*)'

0/50       0/46      1/46(2*)"   15/49(37*)'      3/44(7*)J    1/43(2*)
                                                                                                         3/49(6*)      0/45
                                                                                                         1/10(10X)
                                                                                                         2/50(4X)9    9/50°
                                                                                                         l/SO(2X)k
                                                                                                        10/50(20*)!
                                                                                                    1/36(3X)   0/481
                                                                                                                                  0/8
                                                                                                              0/18
                                                                                                                                   1/50(2X)  0/49
                                                                                                                                                       28
                               1/49(2X)    0/48      1/40|3*)h    18/39(46*)'     4/43(9X)J   7/42(17X)n    1/49(2X)«     1/45«2X)J    0/36      1/48(2X)   36
                                                                4/39(10X)f>«                             2/49(4X)k     5/45(llX)«
                                                                                                        10/49(20*)'
       'Source:  NCI.  1979
       bTotal  -  total antMls with tuaors
       c	Number of antaals bearing tuaor stated	
       NiMber of  anlaals with  tissue stated examined Microscopically
       dNeoplasttc nodule In the  liver of low dose
       eHepatocellular caret now  In the liver of low dose
       *lotal  tumor  Incidence of  tuwtrs In pituitary (p»0.012. Cochran-Arattage test) coapared with pooled  controls
       "Bile duct  haMrtoM
       ^Chroaophobe  ademxu
       JCorttcal adnoaa
       kflbroM
       ^Flbroadenoaa
       Pancreatic llpoaa
       "Eolllcular cell adenoM  Incidence significant (p=0.021. fisher exact test) compared with pooled  controls
       °StroMl polyp
CO

-------
    In  the  case of  mice,  greater  toxldty  of toxaphene was  reported  (NCI,
1979).  As  In  the  case "of rats,  survival  1n  treated  groups  was  high;  90-98%
of  treated  and control animals  survived  beyond the 52nd week of  the  study.
Of  the  tumors  appearing  In  treated  animals,  none were observed  In  greater
Incidence compared  with controls with  the  exception  of those  In  the  liver.
Hepatocellular carcinomas  occurred  1n treated mice only, with  Incidences  of
69% and 98%  In males  (Table  V-10) at  the  low and high doses, and 10% and 69%
In  females   (Table  V-ll)   at  the  low and  high  doses,  respectively.   These
neoplasms were  not observed  In  control animals  of either  sex,  but  hepatic
nodules  were  observed  1n  20%   of  matched-control  males,  though  not  In
females.  On  the basis of these  findings  the authors  concluded  that  toxa-
phene  caused Increases  In  the   Incidence  of  hepatocellular  carcinomas  and
neoplastlc nodules, and hence was carcinogenic In B6C3F1 mice.

    Reexam1nat1on of  the   hlstologlcal  sections  prepared In  the  above  study
led Reuber  (1979)  to  come to qualitatively similar,  but quantitatively dif-
ferent  conclusions.   In this  regard  he reported  liver  carcinoma  Incidences
of 20,  78 and  100% In male  mice  Ingesting  zero,  low or high  doses  of  toxa-
phene and 0,  31  and  89%  respective  Incidences 1n  females.   In  addition one
male mouse  Ingesting  the  low concentration developed  an anglosarcoma  of the
liver, and one female  an adenocardnoma of  the salivary gland.  In addition,
Reuber reported  Incidences of  0,  12 and  31% of stromal cell carcinoma 1n the
uterus  of  females   In  control,  low-dose and high-dose  groups,  respectively,
the  latter  Incidence  being  the  only statistically significant  result.   No
mention was  made of these carcinomas  1n the original NCI (1979) report.
02030                               V-55                             02/26/87

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O
CO
                                                                         1ABIE V 10
                                                 Incidence of  luaors  In Hale Nice  fed loxaphene In  the  Diet'
 I
en
Dose
(•g/kg diet) Initial Examined
0 Hatched 10 10
Low 50 50
High 50 46
Blood Liver Total*
HAL1GNAN1
0/1 Od 2/10(20X) 0
2/50(4X) 6/49(1 2X)e 37
34/49 (69X)r
0/50 45/46(98X)f 45
Lungc

1/10(10X1
1/49(2X)
2/46(4X)
fye
BENIGN
0/10
1/50(2X)
0/50
Totalb

1
2
2
      •Source: NCI. 1979
      bToUl . total an tails with tutors
      cAlveolar/bronchlolar adenoaa
      «•	Nuaber of anlaals bearing iumor stated	
       NiMber of anlMls with tissue stated exaatned Microscopically
      "12K were described Is have "p.ecplasttc nodules.* assuaed to be Ml!«nani. 69X hepatocellular circlnona  and  ?X
      'incidence of hepatocellular  carcinomas  In low and high  dose  groups significant (p<0.001. fisher exact test) compared with Matched or pooled
       controls.
CO

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INJ
o
00
o
                                                                           1ABLE V 11


                                                  Incidence of Tutors In female Mice fed loxaphene In the 0»eta
Dose
(•g/kg diet)
0 Hatched
Low
High
'Source: NCI.
blotal . total
Number of
Initial
10
SO
SO
1979
antMls with
AnlMls
HeMtopolettc Liver Total0 Salivary Mammary Uterus Harder Ian Totalb
Examined System Gland
I
ML1GNAN1 BENIGN
10 0/10* 0/9 0 0/8 0/10 0/10 0/10 1
49 1/50(2X) S/49(10X)d 6 1/46(?X) 1/SO(2X) 2/48(4%) 0/SO 4
13/49(?7X)d
49 0/SO 34/49(b«)*.f 34 0/47 0/SO 0/44 1/50(2X) 1
6/49(1 W)«.f

tumors
c Number of antMls bearing tumor stated
         NuMber «f anlaals with tissue stated exanlned Microscopically


        d27X were reported to have "neoplastlc nodules.*  (ascribed Malignant) and 10X hepatocellular carclnona


        e!2X were reported to have "neoplastlc nodules.*  and 69X hepatocellular carclnotu


        'incidence of hepatocellular carctnotaas significant (p<0.001. fisher exaci  iesij coapared with aatched or pooled tontro! groups
 oo
 tn

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    In addition  to  the previous study, a  number  of  other  much less complete
studies  on  toxaphene  carcinogenic1ty  were conducted.   In a  study conducted
by the U.S. FDA  (Nelson,  1949),  groups of 12 male and 12 female rats assumed
to  be  of  the  Osborne-Mendel  strain   Ingested  0,  25,  100,  400 or  1600  ppm
toxaphene 1n the diet for 107 weeks.   Further  groups  of  5 male and 5 female
rats wore  given 0,  40,  200  or  1000  ppm  toxaphene  and sacrificed  after  56
weeks.    Tissues  were sectioned only  from the first set of  groups,  however,
and Incomplete  histology  was performed on other  groups.  Only  one  neoplasm
was noted,  a  subcutaneous neurosarcoma In a male rat fed the  1000  ppm do-se
level,  although  liver hyperplasla  and hyperplastlc  nodules were reported.
Thyroid hyperplasla was also noted In  treated animals, especially  In males.

    In the  groups  constituting the  107-week experiment,  2/3  female  and  2/2
male rats   Ingesting  1600 ppm  toxaphene  developed  hepatic   carcinomas;  1/3
female rats  In  each  of  the 400  and  1600 ppm  groups  developed hyperplastlc
nodules.   No Incidence of either  of  the above tumors was observed In control
or other  treatment groups (Nelson, 1949).

    A  further  study  demonstrating  a  statistically-significant  Increase  In
hepatic  neoplasms was also  done  (Litton  Blonetlcs,  Inc.,  1978).  Fifty-four
weanling  B6C3F1  mice were randomly  assigned to each  of  four groups.  Toxa-
phene  (X-16189-49),  as well as  the  dose   levels, were selected and provided
by Hercules Inc.  The animals were administered toxaphene  at 0, 7, 20 and 50
ppm levels  1n  the  diet  for  18  months and  surviving  mice were kept  on  the
control  diet  for  another 6  months.   All  survivors  were  killed  24 months
after  the  Initiation  of  treatment.   Table  V-12   shows   the   effects  of
toxaphene on survival.   The  survival  In  both  treated  and  control  groups was
similar.
02030                               V-58                             02/26/87

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                                  TABLE V-12
                  Survival  of  Mice  fed  Toxaphene  1n  the  01eta

Sex


Hale



Female




Dose
(ppra)

0
7
20
50
0
7
20
50

No. at
Start

54
54
54
54
54
54
54
53



1-26
0
0
0
lb
0
0
1
0
Weeks -
1n a Hor

27-52
0
0
1
0
0
0
1
0
Died or Killed
1bund Condition

53-78
1
0
3
0
0
2
2
0



79-105
10
8
6
7
10
7
8
9
aSource: Litton B1onet1cs, Inc., 1978
bM1ss1ng
02030
V-59
02/26/87

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    The  hlstopathologlcal  evaluation  data  presented  1n  Table V-13  shows  a
statistically significant-Incidence  of hepatocellular tumors (hepatocellular
carcinomas and hepatocellular adenomas)  In  male  mice fed  50 ppm toxaphene In
the diet as compared with  the controls.   However,  hepatocellular  tumor Inci-
dence was not statistically significant 1n females as compared with controls.

    Toxaphene was  administered  to  50 weanling ARS Golden  Syrian  hamsters of
each  sex  and each  dose at  levels  of 0,  100,  300  and  1000 ppm  based  on  a
subchronlc  study  (LHton  B1onet1cs,  Inc.,  1979).   The   hamsters,  obtained
from  the Sprague-Oawley  laboratory  In  Madison, HI, were  randomly  assigned to
each  group.   The toxaphene was mixed  1n  corn oil and blended  with  the food
that  was  available ad  libitum.   The  animals  were observed  during  a 15-day
acclimation period  before Initiation  of  the study  and  dally  afterward  for
general condition  and  mortality  Treatment  was  continued for  18  months  for
the females  and  21.5 months for the  males  who showed a  high survival  at 18
months.

    Animals  were  observed  until   spontaneous   death,  unless  moribund,  or
surviving  until  termination of  the  experiment.   Moribund  animals  and those
surviving  until  termination were  sacrificed.   All  animals  were  necropsled.
The postmortem   observations  Included  a  thorough  external  examination  and
collection of major organs and  representative  tissues from all animals.  All
collected  tissues  from  control  and high-dose animals, and target  organs and
gross   abnormalities  from  the  low- and  Intermediate-dose  animals,  were
evaluated  h1stopatholog1cally.   Blood smears were taken  from all  animals at
sacrifice, but were not  examined unless  necessary for definition  of specific
lesions or diseases.  A  compound-related  lowered body weight was  observed 1n


02030                               V-60                             02/26/87

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                                  TABLE V-13



   Hepatocellular Tumor Incidence In B6C3F1 Mice fed Toxaphene In the Diet*
Hepatocellular Tumor Incidence
Dose
(ppm)
0
7
20
50
Hales
10/53 (19%)
10/54 (19%)
12/53 (23X)
18/51 (35X)
Females
2/53
2/53
4/53
6/52
(4%)
(4%)
(8%)
(12%)
*Source: Litton B1onet1cs, Inc., 1978
02030
V-61
02/26/87

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high-dose  males until  12 months  of  study.   Similar  effects were  noted  In
males  receiving 300 ppm "from  months  2-7 and  for  a brief  period  In females
receiving  1000  ppm.   At terminal sacrifice, an  Increase  1n liver  weight was
observed  1n  males  at  the  1000 ppm dose, which  correlated  to the  pathologic
finding  of  megahepatocytes In 6  of  21  males  observed.   A  decrease  In heart
weight  was  noted  In males  at both  the 300  and  1000 ppm dose  levels.   A
decrease In the thyroid gland weights of the 1000 ppm females was also noted.

    Only the  liver  changes were supported  by  hlstopathology  and were there-
fore  judged  to  be   treatment-related.   These  liver  changes  are  similar  to
those  accompanying   an  Induction  of  liver  enzymes  after  administration  of
chlorinated hydrocarbons  and  other  compounds.   Microscopic  evaluation of the
tissues  did  not  reveal  any   Incidence  of  a  tumorlgenlc effect  related  to
toxaphene.   Even when  the  Incidence  of   lymphoretlcular  neoplasms  of  all
types  was  combined, no  difference was  detected between  controls  and dosed
animals.  Under conditions of this study, toxaphene was not carcinogenic.

    It  Is  likely that  the  maximum tolerated  dose was  not  employed  In these
studies  (Litton  Blonetlcs,  Inc.,  1979).    In   the  subchronlc   study,  the
maximum  tolerated  dose  estimate  was  based  on  evidence of  liver  toxldty
only.   No  hamster  at  any dose  level  died during  the  study;  no  changes  In
general  appearance, condition or  behavior among  experimental animals  was
observed;  and  measurements  of  mean  body   weights  and  food  consumption
Indicated no  compound-related change.  In  the chronic  study, a dose-related
decrease  In  body weight was  observed,  but H was  temporary.   The weight  of
treated  animals  equaled or surpassed that  of  controls by  the  end of 1 year
for all  treatment groups.   Treated  animals  also  survived  for a longer period


02030                               V-62                             02/26/87

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 than  controls.   A  comparison  of  the  number  of  animals  1n each  treatment
 group  surviving  to  termination  of  the  experiment  using  an   X2  test  For
 homogeneity  revealed  significant  differences  for both  males   (p<0.05)  and
 females (p<0.001).

    Intragastrlc Injection  of  toxaphene 1n sunflower oil solution  )n  a  dose
 of  50  mg/kg  bw twice-weekly  for  10 weeks  was reported  to give  no  excess
 tumor  Incidence  In  mice  (18 g  Initial  weight)  (Oldenko et  al.,  1978).   The
mouse  strain  was apparently  Indeterminate  (stated  as  nonlinear),  and  both
 sexes  were  used.   Although the  sites  of the  tumors  1n  each  sex  were  not
 reported, the  "tumor formation frequencies"  of  12.5X for treated,  and  8.9%
 for control groups  were reported  to be  not  significantly  different 60 weeks
after  the  start of the  experiment.   Of  toxaphene-treated  mice,  8  bore
pulmonary adenomas,  1  adenocarclnoma,  and  1  lymphogranulomatosls.   In  the
control  group,  five  mice had  pulmonary adenomas.   In  a  parallel  study  In
rats, animals of 80  g weight  were  Injected twice-weekly for  16  weeks  with 80
mg  toxaphcne/kg bw  1n  sunflower   oil.   No  primary  data were  presented  but
toxaphene  Injection was  reported not   to  Increase   the  frequency  of  tumor
formation compared  with controls,  or to  Induce differences  In  tumor  distri-
bution relative  to  gender.  Both control  and  treated  animals  were reported to
develop leukemia and lung tumors by the age of 2.

    In a  study  on  the effects of  toxaphene on spontaneous  and BaP-1nduced
cardnogenesls  In  female A/J mice  (TMolo et al.,  1982), the  pesticide  was
added to  the diet In a  fashion such  that control diets contained 5% corn  oil
and experimental diets  5% corn oil and  100  or  200 ppm technical grade toxa-
phene.  Mice were 9  weeks old  on  Initiation  of  the study, and were  maintain-
ed  on  these  diets   for  12 or  20  weeks.  Following the  12-week  experiment

02030                               V-63                             02/26/87

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essentially  no  tumors  of  any  kind  were  found  In  the  forestomach,  lung,
liver, glandular stomachr  Intestinal  tract  or  spleen  from control  or experi-
mental groups.   When mice were  Intubated with  3  mg BaP  In 0.25 ml  corn  oil
on  days  7 and 21  by oral  Intubation,  100  ppm toxaphene  In  the diet  had no
effect on  BaP-1nduced  forestomach  tumorlgenesls,  but 200  ppm caused a stat-
istically  significant decrease  In  tumor  Incidence.   In  the case of the lung,
200 ppm was without  effect,  but 100  ppm resulted  In decreased tumor  (presum-
ably  adenoma)  Incidence slgnlfIcant.ly compared with  BaP alone.   Concurrent-
ly, the same dietary concentrations  of  toxaphene  were found to significantly
Increase control and BaP-1nduced hydroxylase activities  In the liver,  but to
reduce them 1n lung (TMolo et al., 1982).

    In  the 20-week  portion  of this  experiment  BaP-treated mice  Ingested
toxaphene  at  100 ppm 1n  the diet; a  significant  decrease 1n  the number of
lung  tumors  per  mouse  was   observed  compared with  mice  treated  with  BaP
alone.  In mice treated with the pesticide  alone, the  Incidence of  tumors
was low and  Identical   to  untreated  controls.   The  Incidence of  forestomach
tumors  In  the animals  receiving  BaP  and toxaphene was  reported  to  be  too
large to  quantify  accurately.  Similar effects were  seen on  BaP-hydroxylase
activity  after  20 weeks  of  toxaphene  Ingestlon to  those observed after  a
12-week feeding period  (TMolo et al., 1982).

    In order to  assess  the carcinogenic  activity of  toxaphene In  vitro,  the
effect  of  the  pesticide  at  concentrations  of 0.3-100  yM  on  C3H10T1/2CL8
cell  transformation  Induced  by  4   yM  BaP  was   Investigated  (Nesnow  and
Leavitt,  1979).   Of  a  number of  agents  tested,  toxaphene was  least  active
but was reported to Increase  the  BaP Induced  transformation frequency  In  a
dose-related manner.   No primary data were presented.
02030                               V-64                             02/26/87

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Summary
    Acute  oral  exposure of mammals  and  birds to  toxaphene  resulted  1n.LD5Q
values  ranging  from  7.5-10  mg/kg bw  In  female monkeys  to  75-500 mg/kg  In
rabbits  (see Table  V-l).   The  LD5Q values  resulting  from dermal  applica-
tion  of  toxaphene  are generally  higher  than  those  observed  after  oral
exposure  and range  from -250  mg/kg bw  to  -4000 mg/kg,  both values  being
reported  for rabbits  (see Table  V-2).   Toxaphene  fractions  or  components
differed   1n  chemical   composition,   polarity   and   solubility   leading  to
>10-fold  differences  In toxldty.   The  vehicle  1n  which  the pesticide  Is
solublUzed  for administration  also appears  to  have an effect on  the toxic
response.   Specific  components  of  technical toxaphene are  more toxic  than
the  complex mixture,  and  as  reported  for  mice,  ID,-  values  for  several
components  varied  from  2.5 to  >100  mg/kg bw (see Table V-3).  Brain  levels
of toxaphene may be  Indicative  of acute  toxldty  (see  Table  V-4).   In  swine,
>4 mg/kg  wet weight (brain tissue)  constituted a lethal  level, and 2 mg/kg
was associated with clinical signs of toxldty.

    Subchronlc oral doses of  toxaphene 1n laboratory animals resulted  In few
clinical  signs  of  poisoning.   Responses  varied  from  lethality  In  several
mice given  1280 ppm In  the diet  for  6 weeks to no apparent adverse effects
In rats  following  1ngest1on of  feed containing 189 ppm toxaphene  (see Table
V-5).  At doses <189 ppm In the diet,  subcllnlcal  toxldty was reported such
as  decreased bile  production  and   questionable  liver pathology.   In  mice
receiving 100 or 200  ppm toxaphene  1n the diet,  humoral  antibody  production
(IgG  antibody  formation)   was   suppressed;   however,   cell-mediated   Immune
responses were not  affected.
02030                               V-65                             02/26/87

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    Toxaphenc  aerosols  In  the  form  of  dusts  are  apparently more  toxic  to
rats  than  toxaphene  mists.  M1st  concentrations  as  high  as  500  mg  toxa-
phene/ma  for   3  weeks  caused  no mortality  1n  rats  and  rabbits;  at  12  mg
toxaphene  dust/ma  for   3  months,   mortality   occurred   1n   rats,  dogs  and
guinea pigs.  These results may be a function of particle size.

    Long-term .exposures  of animals   to  dietary  levels  of  toxaphene  are sum-
marized 1n  Tables  V-6 through V-ll.   In  a number of studies  (see Table V-6),
no  adverse effects  were  reported  among  the   parameters  monitored  1n  each
experiment.   In  several  lifetime studies  In  rats,  no effects  were reported
for doses  between  10  and  100  ppm toxaphene 1n the diet;  other studies at 100
ppm  reported   liver  pathology.    The  lowest  dietary  level  producing  liver
damage  In  dogs was  5 ppm  In  the diet  for  2  years.   In addition  to  liver
pathology,  a  variety  of adverse  effects  are reported from  both  J[n vivo and
in  vitro   studies:    kidney  pathology,  decreased   bile  flow,  decreased
survival,   Inhibition  of  Intestinal  transport  of glucose,   Inhibition  of
gluconeogenesls,   Inhibition   of   mitochondria!   enzymes,   Inhibition   of
                   *                                      *5
Na'/K'-ATPase   brain   and   kidney   activity    and   Mg   -ATPase   activity
Inhibited  1n mouse kidney, liver and brain.

    Toxaphene  Induces the  mlcrosomal MFO system, and  treatment  of an animal
Ifl  v^vo  wlth  a  cytochrome  P-450  system  Inhibitor   Increases   toxaphene
toxlclty.

    In  a   3-generatlon  study  with  25 or  100   ppm  toxaphene In  the  feed  of
Sprague-Dawley  rats,  no effects  on parental animals  or  offspring  could  be
detected.   With CD rats, maternal mortality  was 31% at 35 mg toxaphene/kg bw
02030                               V-66                             02/26/87

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and  there was  a  dose  related  decrease  1n  maternal weight  gain  and  fetal
weight  (15,  25 or  35 mg/kg  bw).   No adverse  effects  were reported  In  the
offspring  of  mixed-strain  white  rats  given  4 mg/kg  bw  during  the  entire
pregnancy.  In  hamsters  given the same dose,  the  authors  reported toxaphcne
to be teratogenlc but did not Identify the anomaly.

    More  sensitive  endpolnts  In  rats  were examined by several  Investigators.
Toxaphene  (oral),  at   12  mg/kg   bw  given  to  pregnant  rats,  depressed
chollnesterase  activity  1n  fetal  cardiac   neural   structures  and  delayed
cardiac  neural  differentiation.   At  25  mg/kg bw,  of  the many  biochemical
parameters examined,  the only significant  difference was  noted  in the kidney
(decrease  In alkaline phosphatase  activity and total protein).   There was no
effect  at 12.5 mg/kg bw on  the rat  kidney.   In two  separate  experiments,
there  was  little  evidence  of   behavorlal   deficits   In  the  offspring  of
toxaphene  treated'dams  except for retarded  righting ability  (6  mg/kg and 50
ug/kg bw).

    Toxaphene,  given  to pregnant  mice  (15  mg/kg and  25 mg/kg  bw)  had  no
adverse effect  on parameters  examined 1n offspring.  In  this  same study  the
high dose  (35 mg/kg bw)  Induced  encephaloceles 1n five  Utters  of 90 treated
dams.   A  5-6  generation study  with  toxaphene  (25 ppm)  In the  diet resulted
In  little or  no adverse   effect  In  mice.   No  effects   were  noted  on  the
anatomical development  of  the fetal  guinea  pig from pregnant  females given
15 mg/kg bw from days 21-35 of gestation.
02030                               V-67                             02/26/87

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    Toxaphene  appears  to have a  low teratogenlc potential unless  doses  are
high  enough  to Induce maternal toxldty.  In  the  studies  reported  here,  the
lowest  dose  producing an  effect  was  50  yg/kg  bw  (rats)  given  In  the  diet
from  day  5 of gestation  through  termination of the  study;  effects Included
significant  decreases  1n  overall  swimming  ability  and righting ability  of
young pups when toxaphene was administered before postnatal day 16.

    Toxaphene  Is  mutagenlc   1n  the  Salmonella/mlcrosomal  reverse  mutation
assay using  strains  TA98 and  TA100.   Results of assays of  toxaphene  compo-
nents for  their  mutagenlc potential  Indicated  that mutagenlclty resided  In
the polar  fraction.   Mutagenlclty  was decreased by  the addition  of  active
MFOs, S9;  this agrees  with the in  vivo  observation  that Inhibitors of cyto-
chrome  P-450  Increased toxldty.   The mutagenlc  activity of  commercial prep-
arations varies.

    Studies of toxaphene cardnogenlclty are reported  1n  Tables  V-8 through
V-14.    In  one study with  cardnogenlclty  as the endpolnt  (NCI,  1979),  many
doses were lowered  because of overt  toxldty  1n the  rats  and mice  Ingesting
toxaphene  containing  feed.  The  concurrent  numbers  of  control  animals  were
low (10/sex/group)  and  historical controls  were used  1n  order  to  match  the
number  of  treated animals per group  (50  of each  sex/group).  The  range  of
doses given  to  the  animals  makes  1t difficult to  use THAs  of doses as  a
basis on  which to estimate  risk  for humans  (Table V-14).   It  was  concluded
that, under  the  conditions  of  the bloassay,  toxaphene Is  carcinogenic  In
male  and  female  B6C3F1  mice,  causing  Increased  Incidences  of hepatocellular
carcinomas 1n  a  dose-related  manner.  The results also revealed that toxa-
phene 1s carcinogenic for the  thyroid of male and female Osborrne-Mendel rats.


02030                               V-68                             04/02/87

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                                  TABLE  V-14

              Variation 1n Ooses  of  Toxaphene  Used  In NCI  (1979)
                           Study  of  Cardnogenldty
                                       Time-Weighted
Species              Sex                Average  Dose            Range of Dose
                                       (ppm  In feed)            (ppm In feed)
Rata

Mouseb

male
female
male
female
male
female
1112
556
1080
540
198
99
640-2560
320-1280
640-1280
320-640
160-320
80-160
Increased  Incidence  of  thyroid  tumors  1n  both males  and females  at  the
 high doses.

Increased  Incidence  of  hepatocellular  carcinoma  In  males and  females  at
 both doses.
02030                               V-69                             02/26/87

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In  another  study  with  B6C3F1 jnlce  of both  sexes,  using 7,  20  and  50  ppm
toxaphene In  the  diet,  the cardnogen1c1ty of toxaphene  was  demonstrated  at
these much lower  doses.  After  18  months  of  toxaphene Ingestlon  and 6 months
on  a  control  diet,  there  were  Increased  Incidences  of  hepatocellular
carcinoma 1n males.  The response 1n females  was  less pronounced.

    Reports from  the  U.S.  FDA study (Osborne-Mendel  rats) Included  one  sub-
cutaneous neurosarcoma  In  a  male rat given 1000 mg  toxaphene/kg  diet,  liver
hyperplasla and hyperplastlc nodules, and thyroid hyperplasla.

    A significant  Increase  1n hepatocellular  tumors was  detected  In  mice  In
two separate  studies.   When  adenomas and carcinomas  are  combined,  a  statis-
tically  significant  Increase In  thyroid  tumors  was  noted  In both male  and
female rats along with  a significant  Increase In the cumulative  Incidence  of
pituitary tumors  1n  female  rats.   These findings are  considered  sufficient
evidence  to  classify  toxaphene as  an  animal  carcinogen.   Using the  IARC
criteria  for  classifying  the carcinogenic evidence  with  "sufficient" animal
data,  toxaphene  1s ranked  1n the IARC  2B  group,  meaning that  toxaphene  Is
probably  carcinogenic  1n  humans.   Classification  according  to  the   EPA
guidelines for carcinogens classifies  toxaphene  1n  group B2:   Probable Human
Carcinogen,  meaning   that  there  1s  sufficient  evidence of  cardnogenldty
from animal  studies and Inadequate or no data from ep1dem1olog1c  studies.
02030                               V-70                             04/02/87

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                         VI.   HEALTH  EFFECTS  IN  HUMANS
Acute Tox1c1ty
    Toxaphene, like most  Insecticides,  Is  poisonous  by  swallowing,  skin  con-
tact,  or  Inhalation  (Boots  Hercules Agrochemlcals,  Inc.,  n..d.).  The  fol-
lowing properties  are cited  1n Boots Hercules  Agrochemlcals,  Inc.,  undated.
Emulsions or  solutions  In  vegetable  oils  facilitate  skin  or  gastrointestinal
absorption.   Solutions  1n  kerosene  are  hazardous,  but  absorption 1s  much
more  "erratic."   When toxaphene Is  1n  the form of a powder,  dermal  absorp-
tion  Is  negligible,  but Inhalation  exposure  of resplrable particles may  be
considerable.  Toxaphene poisoning In humans  1s manifested  by  diffuse  stimu-
lation of  the central nervous system resulting 1n  salivation,  restlessness,
hyperexc1tabH1ty,  muscle  tremors  or   spasms,  generalized convulsions,  and
sometimes loss of  consciousness.  Nausea  and  vomiting may  follow oral  1nges-
tlon.  Clonlc  convulsions  may  also  occur and  can  be prevented  by  barbitu-
rates  (I.e.  sodium pentobarbHal) administered  preferably  by  1.v.  Toxaphene
has a  fairly long duration of  action  (I.e.  over several hours)  and a  long-
acting barbiturate  such as phenobarbltal  can  be used after  Initial  control
of convulsions.   In lethal cases, convulsions continue  until  death,  which  Is
ultimately caused by respiratory failure.

    The  IUPAC  (1979)  has  estimated  an  acute oral LD,Q  dose  of  60 mg  toxa-
phene/kg bw.  Hayes (1975) reported  a level  of  78 ppm 1n the liver  of  a  dead
person poisoned  with  toxaphene as  well as  10  ppm In blood.   Conley  (1952)
estimated  an acute lethal dose  to  be   between 2 and  7  g/person  or  29-100
mg/kg for a 70 kg man.
02040                               VI-1                             02/14/85

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    Fatal Intoxications of humans have  been  caused  by  "cotton  dust"  contain-
ing 5X  DOT,  10% toxapherre and  40X sulfur.  The "subacute  fatal  human  dose"
for this mixture Is  reported  to  be  0.75 g  DDT  * 1.5 g toxaphone (Heyridrlckx,
1960).

    Several   cases   of  death  from  toxaphene  poisoning  have  been  recorded
(Hayes,   1963).   A  9-month-old  child poisoned  with a  2:1  mixture  of  toxa-
phene:DOT  died  after   convulsions   and respiratory  arrest.  Toxaphene  was
detected 1n  the liver  (7.9 ppm), kidney (6.8  ppm)  and brain  (14  ppm).   The
ratio of toxaphene:DDT  In  the brain and liver  was  10:1,  and  In the  kidneys,
3:1  (Haun  and  Cueto,   1967).    Four   other  cases  of  acute  poisoning  In
children, 3  of  them fatal, have  been  reported (McGee  et al.,  1952).   Table
VI-1 summarizes  the  findings  of  this study.  No residue  data  were reported.
Recovery  of  humans  from  poisoning occurred  at  doses  of 10-47  mg/kg bw.
Vomiting was  Induced so 1t Is  not clear  how much toxaphene was  absorbed.

    Poisoning by  prolonged skin  contact  with  a Undane/toxaphene  solution
has been  reported  by  Pollock (1958).    The  patient was  semiconscious,  dis-
oriented  and  stuporous,  nauseous  and  had  vomited  many  times.    Severe
epigastric pain,   severe  aches,  and  pains  of the  extremities  were  also
reported. Eventually  stiff neck, dehydration,  and Irregular  pulse  develop-
ed.   Treatment  was  with  penicillin,   chloromycetln  and  digitalis.    The
atypical symptoms  may  be  related to the  presence  of  Undane  or  the  dermal
portal  of  entry.   No  residue  data were  given.   Hayes  (1963)  estimated  a
hazardous dermal dose  to be  46  g.   For a  70  kg man,  this  Is  equivalent to
660 mg/kg or  about 10 times that of an oral dose.
02040                               VI-2                             02/25/87

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o
ro
O
 I
CJ
                                                                              1ABIE VI-)


                                   Case Studies of Toxaphene Poisoning In Humans In which Ingest ton Has the Primary Route of Entry*
Case No.
Sub)ect(s)
Mature of
toxaphene
Dose
Time to react
symptoms
Symptoms

Outcome
Time of death
or recovery
1 2
Male. 2 years Male. 4 years
8 Moths
Max Emulsion In
water
Unknown Unknown
7 hours 2 hours

Convulsions Convulsions
2-5 Minute
Intervals

Death Death
9.S hours 6 hours

3
Hale. 1 year
S Months
60X in
solvents
Unknown
MS

Convulsions
Intermittent

Death
11 hours

4
Male. 2 years
20X In solution
Unknown
MS

Convulsions. Intermittent
•lid cerebral excitement;
aimless jerking motion
and excessive muscular
tensions of extremities.
marked pharyngeal and
laryngeal spasms; labored
respiration; cyanosis
Recovery
12 hours

5
Female. 49 years
Female. 20 years
Female. 16 years
Female. 12 years
Residue of spray
In food
9.5 47 mg/kg
1.5-4 hours

Nausea; vomiting
convulsions

Recovery
12 hours

6
Hale, adult
Male, young
i
Residue of spray In
food
Unknown
4 hours

No nausea; spontaneous
vomiting; convulsions.
Jerking and transitory
movements; muscular
rigidity; periods of
unconsciousness;
amnesla(?)
Recovery
24 hours

           •Source: HeGee et al.. 1952


           NS ~ Not specified
 'CO

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    Allergic  bronchopneumonla  was  Immediately  observed  In  two workers  who
had  just  used  toxaphene, sprays  (Warrakl,  1963).  One  had  been  exposed  to
other  pesticides  for   4  years  and  showed  marked  bilateral  hllar.  lymph-
adenopathy  with  fine mlHary  opacities  heavily distributed over  both  lungs
which,  after   treatment  with  dlhydrostreptomycln  and  1son1az1d,  regressed.
The  other  man also  showed  similar  symptoms,  and no  past  pesticide  exposure
was  evident apart  from that  to  toxaphene.  The  m111ary shadows  regressed
when  the  man  was  treated with streptomycin,  1son1az1d, prednlsone  and  cor-
tisone.   Both  cases evidenced  the  following:  sudden  exertlonal  dyspnea,
tachycardia and  weakness  with  low blood pressure; extensive mlHary shadows.
In  both  lungs;  blood  eos1noph111a; high  serum globulins; reversibility  of
X-ray  shadows,  symptoms and physical  signs to  normal  on  treatment with  cor-
tisone under cover of antltuberculosls drugs;  and acute pneumonia symptoms.

    Eight women  working In  an area that  had  been  sprayed  with  2  kg  toxa-
phene/ha  by aircraft  showed  a  higher  Incidence  of  chromosome  aberrations
(acentric fragments  and chromatld exchanges), as  observed  In  lymphocyte  cul-
tures,  compared  with an  unspecified  number  of control  Individuals:   13.1%
vs. 1.6X (Samosh, 1974).

    The sequelae  observed after  a pesticide fire In  which  -80 fire  fighters
were  potentially  heavily  exposed  to  toxaphene  amongst  many  pesticides
(Mellus and Schulte, 1981)  were  generally  vague (tension,  depression,  hyper-
exc1tab1l1ty,   fatigue,  and  confusion  were significant relative  to the  24*
fire  fighters  who reported  acute symptoms  of nausea,  dizziness,  headache  at
the time of the fire).
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Subchronlc and Chronic Toxlcltv
    Application  of  an aerosol spray  containing  toxaphene to  the  skin  of  50
human  subjects  dally for 30  days  at  a  dose of 300 mg/day  produced  no  toxic
manifestations.   Fifty  human volunteers  who  Inhaled  0.4 mg/m3  of toxaphene^
aerosol  for  10  minutes  a  day  for  15  days had  no subjective  or objective
effects.   Assuming  alveolar ventilation  of  5.4  i/m1nute  the  dally  dose
equalled  -0.02 mg.   A  mist  containing  250 mg/m3  toxaphene was  Inhaled  by
25  humans  for  30 minutes each day (-41  mg/day)  for 13 days  and they showed
no evidence of local or systemic toxic manifestations  (Shelanskl, 1947).

    Many  studies  exist that  Involve long-term  toxaphene exposure to workers.
Most of  them do  not  detail  exposure data, Involve many other pesticides,  and
do not  attempt  to correlate  serum  levels  of toxaphene  or Us metabolites  to
known  symptoms  of toxaphene so  that  the  results,  while suggestive,  are very
often  not  definitive.   Examples of  studies  of this kind follow.   One  was a
health  study  detailing an  Increased Incidence of  lung  cancer  (10  observed
vs. 0.54  expected) among  285 pesticide  applicators (Barthel, 1976).   Another
was a  survey  Involving  321  exposed and  46  nonexposed  males   1n Kuwait  In-
which  the  260  exposed  to   organochloMnes  reported  Increased  sputum  and
hyperreflexla  compared  with  controls  (El-Desouky  et  al.,  1978).   A  third
study was a nonsignificant  case-control  study  of  the  association between  the
yearly  rate  of fatal aplastlc anemia and the  use of many  pesticides In  the
USA between  1950 and 1975  (Wang  and Grufferman,  1981);  a  fourth Investiga-
tion was  a cancer morbidity study  (Barthel,  1981) In  1658 male workers of at
least  5  years  experience as  agricultural  plant  protection  workers  or  plant
protection  agronomists  In  the  German  Democratic  Republic   between  1948  and
1972.  Toxaphene  exposures  occurred after 1954.   An  Incidence  of 169 mallg-


02040                               VI-5                             02/14/85

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nant  neoplasms  was  found.   The ones with  a  significant  excess compared with
the  general   population^were  bronchial  carcinoma,  59  (35%)  to  42  (24%),
respectively, of  which 22  were undlfferentlated and  small-cell  carcinomas,
14 were squamous  epithelium  carcinomas,  3 were  adenocarclnomas,  2  were solid
carcinomas  and  one  was an  alveolar  cell carcinoma.   The  average  latency
period for the  lung  cancers  was calculated to be 17^6 years (6-29  years) and
these  cases  had  been  exposed  on  the  average  for  14+5  years  (6-23  years).
Smoking was  not  a significant  factor  1n  these  cases.   Toxaphene was  thought
not to be a significant factor  1n these Incidences.

    In a  survey  of  199 employees  who worked  or  had worked  with  toxaphene
between  1949 and  1977,  with  exposures  ranging  from 6 months  to 26  years-
(mean 5.23 years), 20  employees died,  one with  cancer of the colon.   None of
these deaths could be attributed to toxaphene (IARC, 1979).

    No Increased  porphyrln  production was  found 1n the urine  of  45  workers
exposed  dally to  a  variety of  pesticides,  predominantly  parathlon,  toxa-
phene, DDT and dleldrln.  The  qualitative method used was  capable  of  detect-
ing  0.4  yg  porphyr1ns/ml.    No   correlations  were  found  between  serum
pesticide  levels  and  urinary  excretion  of  ALA,  PBG  and  CCA.   Parathlon
depressed plasma  or  red cell  ChE or  both In  five workers.   These  findings
suggest  that ordinary  occupational  exposure to the  pesticides noted  above
has no strong porphyrlnogenlc or sympathotonlc effects (Embry et al.,  1972).
02040                               VI-6                             02/14/85

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Summary
    Toxaphene has  caused  convulsions  and nausea  In  humans  when  exposure has
occurred  by  swallowing,  skin  contact  or  Inhalation.   The acute  oral  L0,n
has been  estimated In  one  study  to be 29-100 mg/kg,  and  60 rng toxaphene/kg
bw  In  another.   Mixtures  of  toxaphene  with DOT  (2:1,  by  weight)  are  more
acutely  toxic  than either  component  alone,  the  subacute  fatal  human doses
being 0.75 g  DDT and 1.5 g  toxaphene.   The  liver,  brain and kidney can show
residues.  The  estimated  hazardous dermal  dose  Is -660 mg/kg  bw for liquid
toxaphene.  Allergic  bronchopneumonla has been observed after  acute  Inhala-
tion of toxaphene sprays.

    Dermal doses  of  300 mg/day for  30 days, and  Inhaled  doses  of  0.4 mg/m3
for  10 minutes  on  each   of  15  days,  and  250  mg/ma  for  30  minutes  (-41
mg/day) on each of 13 days caused no atypical subjective or  clinical effects.

    Studies  In  the  workplace  are  confounded  because   exposure  to  many
chemicals occurred 1n all  the  reported studies.   There are  no porphyrlnogen-
1c or sympathotonlc effects mentioned  1n the available literature.

    Human data are Inadequate  to  assess the human carcinogenic potential for
toxaphene.
02040                               VI-7                             02/25/87

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                         VII.  MECHANISMS OF TOXICITY
Introduction            —•
    Toxaphene toxldty  1s  Influenced  by  many  factors:   age,  gender,  species,
state  of  mlcrosomal-enzyme  systems,  presence  of  starvation  or  of  other
chemicals  that  may  affect  cytochrome  P-450  levels,  and the  type  of  carrier
utilized In toxldty experiments (see Chapter  V).

    In  all  cases the  obvious acute  effects  In  humans and  animals  (saliva-
tion,  hyperexcltabUHy,   behavioral   changes,  muscle  spasms,   convulsions)
Indicate that  the central  nervous  system  Is  the  chief target system.   The
mechanism  of  the  effect   1s  still  unknown.   Other  target  organs  affected
acutely  Include  the   kidneys  (cloudy  swelling,  congestion,  renal  tubular
degeneration),  the  liver  (fatty  degeneration,  necrosis,  Inhibition  of  bile
flow  and  biliary function),  the  testes  (decreased  spermatogenesls)  and  the
skeleton  (altered  cartilaginous   structures   In  birds  and  In  guinea  pig
fetuses).

    At  least  In  Swiss-Webster  albino mice  (Saleh  et  al.,  1977;  Saleh  and
Caslda,  1979),  enhanced  acute toxldty  (oral  LD5Q values) was  associated
with  Introduction of  a single chloro-group In the 8-  or 9-pos1t1on  and  to  a
lesser  extent   1n  the  5-exo-pos1t1on.   A  decrease  In  toxldty  for  hepta-
chlorobornanes  was  noted  for single  chloro-groups  In  the 3-exo or  10-posl-
tlons.   For  the nonachlorobornanes,  additional  chloro-groups In  the 3-exo.
10-posltlons  or 1n  the  8,  10-posltlons  caused  compounds   to  be  much  less
acutely  toxic.   The hexachlorobornane produced  by  removal  of a chlorine  at
the 6-pos1t1on  from the heptachlorobornane was not  much more  toxic  than  the
heptachlorobornane Itself.


02050                               VII-1                            02/12/87

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Enzvmatlc Effects
    Khalifa  et  al.  (197tr)  and Chandurkar and  Matsumura (1979a)  found  that
toxaphcne and Toxicants  A and B could be  reductlvely  dechlorlnated or dehy-
drochloMnated quickly  or their vicinal  chloride eliminated  by  liver  mlcro-
somal  preparations  In  vitro  as well  as  nonenzymlcally  by  reduced  hematln
(Khalifa  et  al.,   1976).   Clearly,  certain  toxaphene  components  will  be
metabolized  faster than  others.   |£ vitro metabolism  of toxaphene by  Iso-
lated  mlcrosomal  preparations  from  mice  required  NAOPH  and  relatively
anaerobic conditions  (Khalifa et al., 1976).   The  fact  that  toxaphcne  gave
type  I binding  spectra  with the  hepatic  cytochrome  P-450  of  rats,  mice,
sheep  and rabbits  Is  suggestive  that  toxaphene  components are substrates  for
the  hepatic  mlcrosomal  mixed-function   oxldase  system  (Kulkarnl  et  al.,
1975).  This  Is  also supported by  the following:   the  toxlclty  of toxaphene
can be potentiated  by  plperonyl  butoxlde (Turner  et  al., 1977);  toxaphene
causes shortened pentobarbUal  sleeping  times  In  rats  (Schwabe and Wendllng,
1967;  Trottman  and  Desalah,   1980);  shortened N-methyl  cyclohexenylymethyl
barbiturate  sleeping  times  In rats  have  been  measured   (Ghazal,  1965);
transient decreases   In  (Peakall,   1976)  and  estrone  (Welch  et al.,  1971)
metabolism;  enhanced  mlcrosomal  enzyme activity (Peakall, 1976;  Trottman  and
Desalah,  1980; Pollock  et al.,  1983);  Increases for pentobarbUal  hydroxyla-
tlon  (Trottman and Desalah, 1980),  amlnopyrlne  demethylase  and aldrln  epoxl-
datlon (Pollock et al.,  1983) and  the level of cytochrome  P-450 (Pollock et
al.,  1983)  and  NAOPH  cytochrome  c-reductase  (Trottman  and  Desalah,  1980)
being  dose-dependent.   The "no significant hepatolnductlve  effect" level  for
1.p.  administered  toxaphene  Is between  5  and  25 mg/kg bw  for these enzymes
(Pollock et al.,  1983).
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    Saleh  and Caslda  (1978)  showed  that  the major  "products  of  Toxicant  8
produced  by  rat  liver Tnlcrosomes  under  anaerobic  conditions  were CAS  RN
57981-29-0  and  65620-64-6 In a  2:1  ratio; these products  were  not  produced
under  aerobic  or  anaerobic conditions  1f  NADPH  was  absent.   These  products
were  found directly  1n  fat,  liver  and feces  of adult  male  Sprague-Oawley
rats  as  well  as  CAS RN  64618-63-9  after  a  gavage  dose of 3.1  mg  Toxicant
B/kg  bw.   Chandurkar and  Matsumura  (1979a)  reported  that  In addition  to  a
pathway  requiring NAOPH,  there  appeared  to  be  a  pathway utilizing  gluta-
thlone, as  found  by EC/GC product  analysis and  Inhibitor  experiments  using
3*C1- and  14C-toxaphene,  and  Toxicants B  and  C as  substrates.   In  addi-
tion,  the  NADPH  pathway  metabolized  Toxicant  C   much more  rapidly  than
Toxicant  B.   Glucuronldes,  sulfonates  and  add  hydrolysable  conjugates  were
also  found.   The  addition of  UDP-glucuron1c  acid to  toxaphene/NADPH  system
Increased  the extent  of  metabolism by  8-fold, but not  when   Sesamex  (an
Inhibitor  of  NADPH)  was  also  present,  also showing  probable  Involvement  of
the glucuronlc  add  metabolic  pathway.   The  water  soluble  metabolites  found
when  glutathlone  (GSH)  was  added were thought  to  be  GSHconjugates  formed
Involving  loss of a  chloride.  Chandurkar  and Matsumura  (1979a)  also Identi-
fied the major dechlorlnatlon  product of Toxicant C  to be CAS RN 70459-31-3,
and  4  secondary  alcohols   (hydroxyls  at  C2t  C~,   C^, C,  positions)  and
one primary alcohol.  Oxidation  products  of  Toxicant  B were not alcohols.
Thus,  different  components  In  toxaphene,  not surprisingly,  are metabolized
differently.   Ionic-chlorine Is  the  major excretion product. In  both  feces
and urine  (Crowder and Dlndal,  1974;  Ohsawa et al.,  1975), and small amounts
of  urine  and  fecal  metabolites are   glucuronldes   or  sulfate  conjugates
(Chandurkar,  1977).  The  small  amounts  of  conjugates  may support the  obser-
vation  by Mehendale (1978)  that toxaphene  Inhibits  hepatob111ary function,
at least  In rats.
02050                               VII-3                            02/25/87

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    Mehrle  et  al.  (1979) proposed  that  the skeletal effects  In  birds  could
be explained by a  functional  deficiency  of  vitamin  C 1n the backbone result-
ing from  shunting  vitamin  C to the  liver  to  enable the organism  to detoxify
toxaphene  In  the   liver  by  mixed  function  oxldase  activity,   as  Is  also
proposed  for the "broken back  syndrome"  caused  by toxaphene for  fish (IUPAC,
1979).  Birds  capable  of  synthesizing vitamin C  1n  the liver  (e.g., ravens,
sparrows)  should  not  be  so  susceptible as  those  that cannot (e.g.  ducks,
geese, quail,  turkeys, doves,  pigeons).  This has not  yet  been shown experi-
mentally however.

    DIPasquale  (1977)  showed  that T5  mg toxaphene/kg  bw/day  (day  21  to  day
35  of  gestation)   Increased   collagen-containing  structures  In  guinea  pig
fetuses,  and  this  was  thought  to  be  due  to   a  functional  deficiency  of
vitamin C related  to maternal  mixed-function oxldase Induction.

    How the  vitamin  C  regulatory  shunt  Interacts with  the hepatic  NADPH  and
glutathlone  systems  1n  the  metabolism of toxaphene  1s  still  unknown,  and It
1s unclear  whether there  1s  a link  between  events  1n the liver  and  those
Induced 1n the central nervous system.

    Delchmann  and   KepHnger   (1970)  noted that  pretreatment  of  rats  with
aldrln  or  dleldrln  raised  96-hour  ID.g   values  2-fold,   and 3-fold  after
pretreatment with  p,p-DOT.   Since  these compounds  are all known  to  Induce
hepatic mixed  function oxldase activities,  this may mean that  compounds that
Induce  this  system  will  lower the  acute  toxldty of  toxaphene.   This  was
also so when toxaphene was  administered  together  with  parathlon,  dlazlnon or
02050                               VII-4                            02/12/87

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 trlthion  with  which  toxaphene  Is  often co-formulated (KepHnger and  Dekh-
 mann,  1967).   The  potenilatlon  of  toxaphene  toxlclty  with  piperonyl  butoxlde
 (Turner  et  al.,  1977;  Saleh and  Caslda,  1978)  and  perhaps  the effect  of
 starvation  (Boyd  and Taylor, 1971;  Srebocan  et  al.,  1980a) may  be  mediated
 through  inhibition  of the activity  of  mlcrosomal  enzymes  although  this  has
 not been shown experimentally.

    Sgbacute administration  of   toxaphene  also appears  to affect  enzymatic
 regulation  of  carbohydrate  metabolism  In  young poultry  (Srebocan  et  al.,
 1978,  1980b).   Thus,  pyruvate:COp  Upase  (ADP) and GTP:oxaloacetate  car-
 boxylase   (transphosphorylatlng)   In  cockerel  livers  were   significantly
 decreased  by  doses  >5  mg/kg diet  after  2 weeks of  exposure.   The  authors
 evoked  adrenocortlcal  mediation as  the mechanism.   The  activities of  beef
 heart  mltochondrlal  sucdnoxldase  and  NAOH-ox1dase  were   Inhibited in  vitro
 t>y  330  yM  toxaphene  (Pardlnl  et  al.,   1971).    GuthMe  et  al.   (1974)
 reported  that  at  100  yMr the  active  transport  of  glucose  through  Isolated
mouse  Intestine was  Inhibited.   The  in  vitro  Inhibition of brain, kidney and
          f*
liver   Na/K-ATPases   (IC5Q«   30   WM)   and   o11gomydnsens1t1ve   (icco»
                           H
15  yM)  and  Insensitive  Mg *-ATPase activities  from male  mice also  Impli-
cates  an  Inhibition  of  membrane  transport  systems  (Trottman and  Oesalah,
1979,  1983;  Fattah and  Crowder,  1980).   Subacute blood  chemistry  phenomena
Include:   Increased  serum add  phosphatase,  glutamlc  pyruvlc  transamlnase,
gamma-glutamyl  transpeptldase activities,  and  Increased neutrophll  counts
for dosed mice  (Baumler, 1975).  This  behavior  Is  consistent  with  mild liver
pathology.
02050                               VII-5                            02/25/87

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    The major  enzymlc  effect  caused  by chronic  exposure appears to be unbal-
anced  sugar  metabolism.-At a dose  to  rats  of  35 mg toxaphene/kg bw/day for
6  months  (Gertlg and  Nowaczyk,  1975),  depressed  (17-20%) LOH  activities
occurred  1n  the  liver,  kidney and  serum,  as  well as  decreased  (50%)  serum
alkaline  phosphatase  and  liver  glutamate dehydrogenase.   Peakall  (1979)
showed  that  levels  of pyruvlc  and  lactic  acids In  blood  plasma were  not
affected  even  after 6  months at  a  dose of  2.4 mg  toxaphene/kg  bw/day  and
postulated that  the effects  on  the LOH  system came  from "rats  stressed  1n
some way."   However,  Desalah  et  al. (1979)  found dosedependent Increases  In
liver  mlcrosomal  glucose-6-phosphatase  and  fructosel,6-d1phosphatase after 8
weeks  of  dosing  Sprague-Oawley rats with doses  of 0,  25,  50 and  75 mg toxa-
phene/kg  diet.   This  was  Indicative of changes  In  the process of gluconeo-
genesls.   Alekhlna and  Kuz'mlnskaya  (1980)  found   that  LOH  activities  1n
myocardium  Increased   after  4 months   of  dosing  "white rats"  with 2.7  mg
toxaphene/kg bw/day, and  In the  liver  only  after both oral  and percutaneous
exposure  (9.4  mg/kg  bw/day).   Ishlkawa et al.  (1978)  reported  that a single
l.p. treatments of 40  mg  toxaphene/kg  bw/day caused  decreased plasma choles-
terol  at  day  60  with  no  effects on  plasma  trlglycerldes, liver  weights,
mlcrosomal protein or  cytochrome P-450  In  old  Sprague-Dawley rats.  No dose-
related   effects   were   observed  on  brain  NaVKf-ATPases   or  ollgomycln-
sensltlve and  -Insensitive Mg *-ATPases  for  doses  of  0,  50,  100,  150  and
200 mg  toxaphene/kg  chow for  8 weeks  to  adult  Sprague-Oawley rats, a 30-40%
decrease  being observed  for doses  above 50  mg/kg diet.  Since in vitro tests
showed  dose-effect relationships,  H  was  postulated  that  1n vivo toxaphene
metabolized  before It reached the brain.   This was  evident also for  those
enzyme  activities  In  the  brain  of  male  ICR rats dosed at  0.  10  25  and  50
mg/kg bw/day over  3 days,  though the activities of kidney ATPases except for
02050                               VII-6                            02/26/87

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 the  ol1gomyc1n-sens1t1ve  Mg f-ATPase  were  decreased   In  a  dose-dependent
 manner   as  was   liver-  ollgomycIn-sensitive   Mg2*-ATPase   (Trottman   and
 Desalah,  1979).   Parvu  et  al. (1980) observed  that  4 mg toxaphene/kg bw/day
 for  30  days  to  Wlstar  rats caused I1p1d accumulation In the Hver, depletion
 of free fatty adds, and Increased liver cholesterol.
    These  data  reveal  that  the  major  target  organs  for chronic  toxldty
effects  are,  1n order of  decreasing  Importance:   liver  (probably  by  Imbal-
ances  1n sugar metabolism and ATPase  activity  depression),  kidney (probably
by  kidney  ATPases),  heart  (catecholamlne  metabolism distortion;  probable
ATPase   effects),   the   central   nervous  system  (catecholamlne  metabolism
distortion;  generalized  tremors  1n  chronically dosed  Osborne-Hendel  rats),
and the  Immune  system (premature  aging effects  In  mice and rats).  In  birds,
the skeleton  Is probably  as  Important  a  target  organ as the  liver  (though
this may also  occur  In young  mammals)  because of the Inability of some birds
to synthesize  vitamin C  In the liver  necessary  for  detoxifying the adminis-
tered toxaphene.

Hutagenlc Activity
    Toxaphene,  and  specifically  a  polar  subfractlon,  Is  a  direct-acting
mutagen.  It  Is  not clear  from  the  literature what the Identities are  of the
compounds  1n  the  polar  subfractlon,  or whether,  like most of  the nonpolar
components, they are metabolized  so quickly  that  In  fact  metabolites  are the
direct-acting  mutagens.    Clarification  of  this  mechanism  Is   an  Important
research need.   Toxaphene has  been  found to be carcinogenic  1n  two  animal
species,  Osborne-Mendel   rats  and  B6C3F1  mice  In  studies  described  In
Tables  V-8  to V-ll (NCI,  1979).   The molecular events  and  the  role  of the


02050                               VII-7                            02/26/87

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polar fraction have  not  been formulated.  While rats  tend  to show malignan-
cies of the endocrine  system  (the  thyroid  and  the  pituitary 1n females) mice
showed cancers  only  In  the  liver.  This  does  suggest the  link  between  the
liver and central nervous system could be by effects on the endocrine system.

Summary
    Figure VII-1  summarizes  the mechanisms  underlying the  acute  to chronic
effects of toxaphene.
02050                               VII-8                            02/25/87

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                            f»t
                            t^Cftl
            ton-polar cMponwu
            *.|. TMlCMIt* M.C
                                                 I.   C«trt1 iwovi tjtlM tffKU
                     t.  (IdMf
                     1.  lt»» 4

                        taut
                     I.
                                f«t«M»
                                                                      *•

                                                                 tak*-*.^
                                                  [ tlnUttloM c«n>n»U» j-
r»»t w
     UkOl
  llwr*
                            (rmlrw .
                            N«iO. »1U«1n C
                                 Mm
teduert k«Mt1n
froducts M t r««
rtduetta* dtdil
                                                                   M •
                                                             dtdilortattlM,
                                                          dilwld* «1talMt1on,
                                                           *    l«i. kjrdr«jrlttt»t
                                      NtU6o11Ui «ur««
                                      «• f»t
                                   k1l« flw k«t
                                      jdviulv
                             ftft •llBlMtlW (UK-
                             lift* »1tt1« 1 M* 111
                             MMr»1 ip*rt trm 1« ktrdi).
                             Cklvld* 1» «J» ncrtttw
                                                         ulfttM vt i
                 T
                       S*«prt«
                               ATTutt (MdXMt
                               (rmturt mtM)
                               Oltttrt*
                                                      SMkdirwIc/dirwU tfTtcU
            (§Ot1d»t1f1idJ_
                            Dtr«et-4CtlH
                                                   1.
                        Wt tff*ct«4 *.|. tfrv,
                        ktrt, e«i»4l iwe«t tyttn.
                        lyitM, kltfnty, MdecrlM ifiitt* (T)
                           l* t«IUB. AtUtOi
                               C«nc*rt AMTVM
                               nuiunr
                               f»ti)
                                    tol«)
                               tolMtU i
                            ICMCV M t Mtol
                            tgCltt «tf <• M
                                       FIGURE  VII-1

     Summary of  Mechanisms Underlying Acute  to  Chronic Effects of  Toxaphene

a!0  mg/kg  diet  for  sheep,   steers,  cows;  4.74 ing/female ring-necked  pheas-
 ant/day;   12.5  mg/kg  bw/day  for   female  White Leghorns; 21 mg/kg  diet  for
 rats,  2.4  mg/kg  bw/day causes  residue  steady state 1n  liver  after 1  month
 and 3 months for  brain

bNo  significant  Inductive   effect  on  affected  liver  enzymes  1s  between   5
 and 25 mg/kg bw for rats.
cl-3 mg/kg bw/day

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              VII-9
                            07/26/84

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                VIII.  QUANTIFICATION OF TOXICOLOGICAL EFFECTS

Introduction
    The  quantification  of  toxicologlcal  effects of  a chemical  consists  of
separate  assessments of  noncarclnogenlc  and  carcinogenic  health  effects.
Chemicals  that  do not  produce  carcinogenic  effects  are  believed to  have  a
threshold dose below which  no adverse,  noncarclnogenlc health  effects  occur,
while carcinogens are assumed to act without a threshold.

    In  the  quantification   of   noncarclnogenlc  effects,  a  Reference  Dor
(RfD),  [formerly  termed  the  Acceptable Dally  Intake (ADI)]  1s  calculated.
The RfD  1s an estimate  (with  uncertainty  spanning perhaps an  order  magni-
tude)   of a  dally  exposure  to the  human  population (Including  sensitive
subgroups) that  1s  likely  to be  without an  appreciable  risk  of deleterious
health effects  during  a  lifetime.   The RfD  1s derived  from  a  no-observed-
adverse-effect   level   (NOAEL),   or   lowest-observed-adverse-effect   level
(LOAEL),  Identified  from a  subchronlc  or   chronic  study, and divided  by  an
uncertainty factor(s)  times a  modifying factor.   The RfD  1s calculated  as
follows:
     IfD . 	'"0>a °r L°"L'	«,A9
           [Uncertainty Factor(s) x Modifying Factor]   	
    Selection of the uncertainty  factor  to  be  employed  In the calculation of
the RfD  1s based  upon  professional judgment,  while considering  the  entire
data  base  of  toxicologlcal  effects for  the  chemical.   In order  to  ensure
that  uncertainty  factors are  selected  and applied  1n  a  consistent  manner.
02060                                VIII-1                          02/26/87

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 the  U.S.  EPA  (1986a)  employs  a  modification  to the guidelines  proposed  by

 the National Academy of-Sc1ences  (NAS, 1977, 1980) as follows:


 Standard Uncertainty Factors (UFs)

        Use a  10-fold  factor when extrapolating from valid experimental
        results from studies using prolonged exposure to average healthy
        humans.   This  factor  1s  Intended  to account for  the variation
        1n sensitivity among the members of the human population.  [10H]

        Use an  additional  10-fold factor when  extrapolating  from valid
        results  of  long-term  studies  on  experimental  animals  when
        results of  studies of  human  exposure  are not available  or  are
        Inadequate.  This  factor  1s Intended to  account  for  the uncer-
        tainty  1n  extrapolating  animal  data   to the  case  of  humans.
        [10A]

        Use an  additional  10-fold  factor  when  extrapolating  from  less
        than chronic  results  on  experimental  animals when there  Is  no
        useful  long-term  human  data.   This  factor  1s  Intended  to
        account  for the  uncertainty  In extrapolating  from  less  than
        chronic NOAELs to chronic NOAELs.  [10S]

        Use an  additional  10-fold  factor  when  deriving  an  RfD  from a
        LOAEL  Instead  of  a NOAEL.   This factor  Is  Intended  to account
        for  the  uncertainty   In  extrapolating   from  LOAELs  to  NOAELs.
        [10L]

Modifying Factor (MF)

        Use  professional  Judgment  to  determine  another  uncertainty
        factor  (MF) that Is  greater  than zero  and less  than or equal  to
        10.   The  magnitude  of  the  MF  depends  upon the  professional
        assessment  of  scientific  uncertainties of  the  study  and  data
        base not  explicitly treated above, e.g.,  the  completeness  of
        the overall  data  base and the  number  of species  tested.   The
        default value for the MF  1s 1.
    The  uncertainty factor  used  for  a  specific risk  assessment  Is  based

principally  upon   scientific   Judgment   rather  than  scientific  fact  and

accounts  for   possible   1ntra- and  Interspedes  differences.   Additional

considerations  not  Incorporated  In  the  NAS/ODW  guidelines  for  selection of

an  uncertainty factor  Include the  use  of  a  less  than  lifetime  study  for

deriving  an RfD,  the  significance of  the adverse  health  effects  and  the

counterbalancing of beneficial effects.
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VIII-2
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    From  the RfD,  a Drinking  Water  Equivalent  Level  (DUEL) can  be calcu-
 lated.   The  DUEL   represents  a  medium  specific  (I.e.,  drinking  water)
 lifetime  exposure  at which  adverse,  noncarclnogenlc health  effects  are not
 anticipated  to  occur.   The  DUEL  assumes  100% exposure  from  drinking water.
 The DWEL  provides  the noncarclnogenlc health  effects  basis  for  establishing
 a  drinking  water  standard.   For  Ingestlon  data,  the  DWEL  1s derived  as
 follows:
                       (RfD)  x (Body weight  In kg)             /B
                      —    	                  — =  	 mci/8.
                      Drinking Water Volume  In i/day   	  "
where:
        Body weight = assumed to be 70 kg for an adult
        Drinking water volume = assumed to be 2 i/day for an adult

    In addition  to the RfD  and  the DWEL, Health  Advisories  (HAs)  for expo-
sures  of  shorter  duration  (1-day,  10-day  and longer-term)  are  determined.
The  HA values  are used  as  Informal  guidance  to municipalities  and other
organizations when  emergency spills or contamination  situations  occur.   The
HAs are calculated using an  equation  similar to  the  RfD  and DWEL; however,
the NOAELs  or  LOAELs  are  Identified from acute or  subchron'ic  studies.   The
HAs are derived as follows:
                   HA   (NOAEL or
                          (UF) x (	 i/day)      	

    Using the above equation,  the  following drinking water HAs are developed
for noncarclnogenlc effects:
    1.  1-day HA for a 10 kg child Ingesting 1 I water per day.
    2.  10-day HA for a 10 kg child Ingesting 1 I water per day.
    3.  Longer-term HA for a 10 kg child Ingesting 1 l water per day.
    4.  Longer-term HA for a 70 kg adult Ingesting 2 l water per day.

02060                                VIII-3                          02/26/87

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    The  1-day  HA calculated  for  a  10  kg  child  assumes  a  single  acute
exposure  to  the chemical and  1s  generally  derived from a  study  of <7  days
duration.  The  10-day  HA assumes  a limited exposure period of 1-2  weeks  and
Is generally  derived  from a study of <30 days  duration.  The  longer-term HA
Is  derived for  both  the  10  kg  child and  a  70 kg  adult and  assumes  an
exposure  period  of  ~7  years  (or  10% of  an  Individual's  lifetime).    The
longer-term  HA   Is  generally  derived  from a  study of  subchronlc  duration
(exposure for 10X of animal's lifetime).

    The U.S.   EPA categorizes the  carcinogenic potential of a chemical,  based
on the overall we1ght-of-ev1dence, according  to  the following scheme:

        Group  A: Human  Carcinogen.   Sufficient   evidence  exists  from
        epidemiology  studies   to  support  a  causal association between
        exposure to the chemical  and human cancer.
        Group  B: Probable  Human  Carcinogen.   Sufficient  evidence  of
        carclnogenlclty  1n  animals with  limited   (Group  81)  or  Inade-
        quate (Group 82)  evidence 1n humans.
        Group  C:  Possible  Human   Carcinogen.    Limited   evidence  of
        cardnogenldty 1n animals 1n the  absence  of  human data.
        Group 0:  Not  Classified  as to  Human Cardnogenldty.   Inade-
        quate human and  animal evidence of cardnogenldty or  for  which
        no data are available.
        Group  E:  Evidence   of   Noncardnoqen1c1ty  for  Humans.    No
        evidence  of  cardnogenldty  In   at  least  two  adequate  animal
        tests  In different  spedes  or 1n  both  adequate ep1demtolog1c
        and animal studies.
    If toxlcologlcal evidence  leads  to the classification of the  contaminant
as a  known,  probable or  possible human  carcinogen,  mathematical  models  are
used  to   calculate  the  estimated  excess  cancer   risk  associated  with  the
Ingestlon  of  the  contaminant  In  drinking water.   The  data  used In  these
02060                                VIII-4                          02/26/87

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estimates  usually  come  from  lifetime  exposure  studies  using animal.   In
order  to  predict  the MsV for humans  from  animal data,  animal  doses  must be
converted  to equivalent human  doses.   This  conversion  Includes  correction
for  noncontlnuous  exposure,  less  than lifetime studies  and  for  differences
1n  size.   The .factor  that  compensates  for  the size difference Is  the  cube
root of  the  ratio of the animal and  human  body weights.   It  Is assumed  that
the  average  adult  human  body weight 1s 70  kg and  that the  average  water
consumption of an adult human 1s 2 l of water per  day.

    For  contaminants  with  a  carcinogenic  potential,   chemical  levels  are
correlated with  a carcinogenic  risk  estimate  by  employing a  cancer  potency
(unit  risk)  value  together with  the  assumption  for lifetime  exposure  from
Ingestlon of water.   The  cancer  unit  risk  1s  usually derived from  a  linear-
ized multistage model with a 95X  upper confidence  limit providing  a low  dose
estimate; that  Is,  the true risk  to humans, while  not  Identifiable,  Is  not
likely  to exceed   the  upper  limit   estimate  and,   1n  fact,  may   be  lower.
Excess cancer risk  estimates  may  also be calculated  using other models  such
as  the one-hit,  Welbull,  loglt  and  probH.   There  Is  little basis  In  the
current  understanding of  the  biological  mechanisms Involved  1n   cancer  to
suggest that any  one  of these models  Is  able to predict risk more  accurately
than any other.   Because each model  Is based upon  differing assumptions,  the
estimates derived for each model can differ  by several orders of magnitude.

    The  scientific  data  base used to calculate  and support  the  setting of
cancer  risk  rate  levels  has  an  Inherent  uncertainty  that  Is  due  to  the
systematic and random errors In scientific  measurement.   In  most  cases,  only
studies  using  experimental  animals  have  been performed.   Thus,  there  1s


02060                                VIII-5                          02/26/87

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uncertainty  when  the  data  are  extrapolated  to  humans.   When  developing
cancer  risk  rate levels,' several  other  areas of uncertainty  exist,  such  as
the  Incomplete  knowledge concerning  the health  effects  of contaminants  In
drinking  water,  the  Impact  of   the  experimental  animal's  age,  sex  and
species,  the  nature  of  the  target organ  system(s) examined  and  the  actual
rate of  exposure  of the  Internal  targets In  experimental  animals  or  humans.
Dose-response data  usually   are available  only  for  high  levels of  exposure
and  not  for  the lower  levels  of  exposure closer to where a standard  may  be
set.   When  there  1s   exposure  to  more than   one  contaminant,  additional
uncertainty results  from  a lack of  Information  about possible  synerglstlc  or
antagonistic effects.

Noncardnoqenlc Effects
    The  effects  of  acute exposure to  toxaphene,  In  humans as  In  animals,
Include  salivation,  hyperexc1tab1l1ty,  behavioral  changes, muscle  spasms,
and  convulsions,   Indicating  that  the  CNS  1s  the  critical  target  organ
system.  In contrast,  the liver and  the  Immune  system appear to be the crit-
ical target sites In  chronic or subchronlc  exposure.  Central  nervous  system
effects  characteristic  of acute  Intoxication appeared during  chronic  expo-
sure only at dietary  concentrations greatly  1n  excess  of  those causing liver
damage In rats (Kennedy et a!., 1973).

    Studies of  the  effects  of long-term  exposure  of workers  to  toxaphene
have been  reported.  These   are of  little  utility  for standard development,
however,  because  exposure generally was  poorly  defined  and often was  to a
mixture  of  pesticides,  and   because no  effort was  made  to  correlate  effects
with plasma or tissue levels of toxaphene or  Us metabolites.


02060                                VIII-6                          02/26/87

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    A  few  controlled  subacute studies of human  toxaphene  exposure  have been
reported.  None  of  these-resulted In any observed  toxldty.   Application of
an aerosol spray  containing  toxaphene  to  the  skin of 50 human subjects dally
for 30 days at a  dose of 300 mg/day produced no toxic manifestations.  Fifty
human  volunteers  who  Inhaled  0.0004 mg/i  of  toxaphene  aerosol  for  10
minutes/day for 15  days  had  no subjective or  objective effects.   Twenty-five
humans  Inhaling  a  mist  containing  0.25  mg   toxaphene/i  of  air  for  30
minutes each  day for  13 days showed  no  local  or  systemic  toxic  manifesta-
tions (Shelanskl, 1947).

    Most acute  studies  to  date  have  Involved  attempts  to define  the LD,Q
for a  variety  of  species.  Lackey (1949a) also  determined the threshold for
Initiation of  convulsions In  dogs.   Single  doses  of toxaphene were admin-
istered In corn oil by  stomach tube  at doses  varying from 5-50 mg/kg bw In 5
mg/kg .Increments.   No convulsions or mortality occurred  In  three  dogs dosed
with 5 mg/kg.  Convulsions  developed In  4/5  exposed to  10 mg/kg.   Mortality
was first  detected  at  the  15 mg/kg  dose (2/8)  with  6/8  developing convul-
sions.  The  LD5Q was  estimated   to  be  -25 mg/kg  bw.   If  repeated  doses  of
toxaphene at 5 mg/kg  bw  were  given,  convulsions  would develop regularly, but
only after several  days.  Four dogs dosed dally  with  4  mg/kg  bw,  2 dogs for
44 days and 2 for 106 days,  survived although convulsions occurred occasion-
ally.   In  the subchronlc  exposures, hydropic degeneration  was  seen  In the
liver with degenerative changes 1n the tubular epithelium of the kidneys.

    Olson  et  al.  (1980)  reported effects at much  lower  doses.   Behavioral
effects were  noted  1n offspring  of  rats  given dally  doses  of  50  yg/kg from
day 5 of gestation  until  70-90 days  postpartum.   Since the effects were gen-
erally transient  and  appeared  to  result  from  temporary delays In maturation,

02060                                VIII-7                          02/26/87

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 It  Is  uncertain whether they can be  classified  as  adverse.   Secondly,  since
 the  offspring  were  expoied  In  utero, by  nursing  and by  toxaphene  added  to
 the  feed,  1t 1s  uncertain  what  actual exposure levels were.   Studies such as
 those of Shaw (1947) Indicate that toxaphene may be concentrated 1n the milk.

     There  are  a number of  subchronlc and chronic  studies of  toxaphene tox-
 Iclty  1n  animals.   Many of  these document changes  1n  activities  of  various
 enzymes, such as  lactate  dehydrogenase Isozymes,  serum alkaline phosphatase,
 and  tissue  ATPases  (Gertlg and  Nowaczyk,  1975; Oesalah  et al., 1979;  Trott-
 man  and  Desalah,  1979; Alekhlna  and  Kuz'mlnskaya,  1980).  Although  some  of
 these observations  are suggestive of  an effect on  gluconeogenesls,  they  are
 of doubtful significance since  there  1s  no direct  proof that  glucose metabo-
 lism Is actually altered, at least In rats (Peakall, 1979).

    There  1s   some   evidence  that  suggests  that   Upld metabolism  may  be
 affected  by toxaphene  exposure.   Ishlkawa  et  al.   (1978)  reported  that  a
 single  l.p.  treatment  of  old  Sprague-Oawley  rats  with 40  mg toxaphene/kg
 bw/day  resulted  In  a  decrease  In plasma  cholesterol  60 days  later  with  no
 effect on plasma tMglycerldes  or on  liver weight.   On the other hand,  Parvu
 et al.  (1980)  observed  that  administration of  4  mg/toxaphene/kg bw/day  to
Ulstar rats for 30  days caused  llpld  accumulation  In the liver, depletion of
 free fatty adds, and an Increase In  liver cholesterol.

    Recently,  Allen et al.  (1983) concluded  that  the  Immune  system  Is  at
 least as  sensitive   to  the Ingestlon  of  toxaphene as  the liver.   Toxaphene
 was  added  to the  diet of  female  Swiss Webster mice at concentrations  of  10,
 100  and  200 ppm  for  8 weeks.    Among groups  of  23-26 animals each,  liver
 weights were significantly  (p<0.05) Increased  1n  the 100 and  200 ppm groups.

 02060                                 VI11-8                          02/26/87

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Among groups of  10  animals  each,  IgG  antibody  tHers  were also significantly
(p<0.05) decreased  at  the  two  higher  doses.   Cell-mediated  Immune responses,
however, were  not  affected.   Offspring of females fed  toxaphone  In  the diet
from 3 weeks before mating  until  weaning  were  also tested at 8 weeks of age.
The cell-mediated  Immune response was  suppressed  1n  the offspring of the 100
ppm  group,  while  the phagocytlc  ability of  macrophages was  significantly
reduced  even   In  the   10  ppm  group.   The actual  toxaphene  dose cannot  be
estimated  accurately   In   the   latter  study   since  the  offspring  received
toxaphene transplacentally,  1n the  milk  and  possibly  even  In  the feed.   It
does suggest,  however, that neonates may be at greater  risk than adults.

    In most studies the critical target  organ In rats  and  dogs  chronically
exposed  to  toxaphene  was  the  liver.   At  dietary  concentrations of  100  ppm
toxaphene In  the  diet  and above,  all  studies document  some form  of  liver
pathology In- exposures  lasting from  2  years  (In  rats  and dogs)  to  lifetime
(In rats).   Toxaphene at 25  ppm 1n  the diet  fed to rats over their lifetimes
caused  an   Increase  1n  liver   weight  with  minimal  liver  cell  enlargement
(FUzhugh and  Nelson,  1951).   Over  shorter periods of  time  (up to 12 weeks),
dietary  toxaphene  levels of  up to 189 ppm had  no  measurable effects on male
or female albino rats  (Clapp et al., 1971).

    The  NCI  (1979) conducted  a chronic  study  with  Osborne-Mendel  rats  and
B6C3F1 mice to  determine  the possible cardnogenldty  of  toxaphene  added  to
the diet.  Each  treatment group consisted of  50 rats  or mice of each gender,
and 10 untreated animals of  each  gender were  matched  controls with data from
40  or  45 untreated animals from similar bloassays  pooled  for  statistical
evaluations.

02060                                VIII-9                          02/26/87

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    Groups  of  50  mice  of  each  gender  were  administered  toxaphene  for  80
weeks  and observed  untTT sacrifice  at  90-91  weeks  (NCI,   1979).   Low-dose
males  and females  received 160 ppm  In  the diet for 19 weeks  followed by 80
ppm for  61  weeks;  high-dose males and  females  received  320  ppm for 19 weeks
followed  by 160  ppm  for  61  weeks.   The  TWA doses were 99 or 198 ppm for both
males and females.   Mean  body  weights of  high-dose male mice were lower than
those  of  matched controls but  weights of  other  dose  groups  were essentially
unaffected  by toxaphene.   Several  animals  died before  week  19 when doses
were lowered.  Following  this,  the  dosed  mice were generally comparable with
controls  In  appearance  and behavior  during  the  first  year  of the  study.
During  the   second   year,  abdominal  detention was  observed  In   all  dosed
groups,  but  predominantly  1n  the  high-dose males.   Other  clinical  signs
Included  alopecia,   diarrhea,  rough  hair  coats,  and  dyspnea.  From weeks
60-76  the low-dose males appeared hyperexdtable.  After week 75  there were
dose-related differences  1n survival.

    Groups  of 50  rats  of  each  gender  were  administered  toxaphene  for  80
weeks  (NCI,  1979)   and  then  observed  until  survivors  were   sacrificed  at
108-110  weeks  (see  Table  V-7).   Low-dose males  were  given 1280 ppm  in the
diet for  2  weeks, 640  ppm for  53 weeks  and 320 ppm for 25 weeks for a TWA of
556 ppm.   High-dose  males  received  2560  ppm  In  the  diet for  2  weeks, 1280
ppm for  53  weeks and 640 ppm  for  25 weeks for  a  TWA  of  1112 ppm.   Low-dose
females  received 640 ppm 1n the diet for  55 weeks and 320  ppm for 25 weeks
for a  TWA of  540 ppm.   High-dose  females  received 1280 ppm for 55  weeks and
640 ppm for 25 weeks for a TWA of 1080 ppm.
02060                                VIII-10                         02/26/87

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    Mean body  weights  of the low- and high-dose  female  rats  were lower than
those of the  matched controls throughout most of  the  study,  whereas weights
of  low-  and  high-dose males  were  essentially unaffected.  During  the  first
16 weeks,  dosed  males  were generally comparable with  controls  in appearance
and  behavior,  with  the  exception  of  high-dose males  that  appeared  hyper-
active during  week  2 when  the doses for  male rats were lowered.  At week 53,
the concentration of toxaphene  In  the  diet  was  reduced because a majority of
the  high-dose  males and  females  developed  generalized body  tremors.   Dose-
related  decreases  In  survival  rates  were  not  observed.   Clinical  signs
usually  associated  with  aging  were observed earlier  In dosed  rats  than  in
controls:  alopecia,  diarrhea, dyspnea,  pale mucous  membranes, rough  hair
coats,  dermatitis,  ataxla,  leg paralysis,  eplstaxls,  hematurla,  abdominal
distentlon and vaginal  bleeding.   Two  females, one  high-dose and  one  low-
dose, had  Impaired equilibrium.

    In  summary,  chronic   feeding  studies suggest  that  there are  no  marked
species differences  In sensitivity among rats,  mice and  dogs,  although mice
may be  slightly  more sensitive than rats to  frank effects  of toxaphene.  On
chronic  exposure,  a dietary  level  of about  25 mg/kg  diet  appears to  be  a
NOAEL,  and a  dietary level  of  40 mg/kg  diet  to  be  a LOAEL, with  liver
pathology  being  the  critical  toxldty  endpolnt.   Chronic  dietary  levels
above  100  mg  toxaphene/kg   diet  are  likely to  be  associated with  frank
effects.
02060                                VIII-11                         02/26/87

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Quantification of Noncardnoqenlc Effects
    Derivation  of  1-Dav  HA.   While  kidney  and  liver  pathology  as well  as
Immune effects are the  critical  endpolnts  for  chronic  exposure to toxaphene,
the  nervous  system  Is  generally  more  sensitive  to   acute   exposure.   The
available data  suggest  a  rather  sharp threshold  for nervous  system effects.
Lackey (1949a)  reported convulsions 1n dogs exposed dally  to  toxaphene  at 5
mg/kg/day after several days, while 4 mg/kg/day  Induced few convulsions  even
with  much  longer exposures.   No convulsions  were Induced  \r\ dogs after  a
single  dose  of  5  mg/kg  bw, while  10  mg/kg  Induced  convulsions  1n  4/5
animals.   The Lackey (1949a)  study  Is the  only one available that accurately
defines  a   NOAEL  for  very short-term exposures.  Dogs  appear  to be  more
sensitive  to the  acute  toxic  effects  of  toxaphene  than  other  laboratory
species  (see Table  V-l)  with   reported  t-D50s  as 1ow as  20-25 mg/kg  bw.
There  1s  little evidence  that  humans are  more sensitive  to  toxaphene  than
dogs.  IUPAC  (1979)  has  estimated  an oral L05Q  dose  1n  humans  of  60  mg/kg
bw, while  Conley  (1952)  estimated  the acute lethal dose In  humans  to  range
from 29-100 mg/kg.  Finally,  the  nervous  system 1s the critical endpolnt for
acute  toxldty  of  toxaphene.   There  1s  no evidence from the  Lackey (1949a)
study  or from the variety  of teratology studies  conducted  that  very  short-
term  exposures  at  the  threshold level  for  convulsions result 1n  kidney  or
liver  pathology.  Using the  NOAEL  from  the Lackey (1949a)  study,  the  1-day
HA Is derived as follows:
                  1  day HA child  =                  a  °'5  mg/l
where:
        5 mg/kg = NOAEL for a single oral exposure to dogs (Lackey, 1949a)
        10 kg   = weight of protected Individual (child)

02060                                VIII-12                         02/26/87

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        100     = uncertainty factor appropriate for use with a NOAEL  from
                  animal data and to protect sensitive members of the
                  human population
        1 i/day = assumed volume of water consumed dally by a 10 kg child
It 1s recommended that 0.5 mg/i for children be accepted as the 1-day  HA.

    Derivation of  10-Day  HA.  The  majority  of subacute exposures  have  been
conducted to  evaluate the reproductive or teratologlc  effects  of  toxaphene.
Chernoff  and  Carver  (1976)  reported  maternal   toxldty  of rats and mice  at
all doses  used (15-35  mg/kg)  during  exposure  from days  7-16  of  gestation.
Kavlock  et  al.  (1982)  found  Increased  total   protein  In  fetuses  from  dams
exposed  to  12.5  and  25 mg/kg/bw on days 7-16  of gestation  In  rats.   Dally
doses  of  12  mg/kg  In  rats  on days  1-2  and  6-14  of gestation  resulted  In
smaller  litters   and  decreased  chollnesterase  activity   along  myocardlal
vessels  and  cells   of   the  atrloventMcular   node   (Badaeva,  1979,   1981).
Toxaphene administered  by  Intubation 1n  doses  of  4  mg/kg/bw to rats  and
hamsters  on  days  7-11 or  1-15  of gestation had  no  effect upon development,
fetal   weight,  ratios of  males  to  females, the  number of  fetal  deaths  or
maternal  toxldty  (Martson and Skepel'skaya 1980b).  The  NOAEL for maternal
toxldty  as  well as  standard reproductive  and  teratologlcal  parameters  in
laboratory animals  appears  to  be 4 mg/kg bw.   Results  of  the Lackey  (1949a)
study,  however,   suggested the NOAEL  may  be slightly  lower  In  dogs  than  In
rodents.  Minimal  kidney  and liver  pathology  were reported  In  dogs  exposed
to 4 mg/kg bw/day for up  to  44 days.   It Is uncertain 1f these effects occur
with  10 days of  exposure.   Occasional convulsions  were also  noted  at  this
dose level.
02060                                VI11-13                         02/26/87

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    Using  a  LOAEL  of  4 mg/kg  bw/day  from  the  Lackey  (1949a)  study,  the
 10-day HA  1s  derived as  follows:
                 10-day HA child = 4 ""I71"* x 10 ^ - 0.04
                                     1000 x 1  I
where:
        4 mg/kg = LOAEL from oral exposure to dogs (Lackey, 1949a)
        10 kg   = weight of protected Individual (child)
        1000    = uncertainty factor appropriate for use with a LOAEL from
                  animal data and to protect sensitive members of the
                  human population
        1 l     = assumed volume of water consumed dally by a 10 kg child
    It  Is  recommended that the  10-day  HA of 0.04 mg/l  for  children derived
from the Lackey (1949a) study be accepted.

    Derivation of  Longer-Term HA.   There are  no  acceptable studies  in  the
available literature for the derivation of a longer-term HA.

    Assessment of  Lifetime Exposure and  Derivation  of  a  DMEL.   There  are
no acceptable  studies 1n  the available  literature  for the  derivation  of  a
lifetime OWEI.

Carcinogenic Effects
    Although several  epidemiology  studies have  been  conducted  attempting to
correlate  cancer   Incidence   with  exposure  to  organochlorlne  pesticides
(Barthel, 1976,  1981;  El-Oesouky  et  al., 1978; Wang  and  Grufferman,  1981),
toxaphene was  not  thought to  be a significant factor  1n the results  of any
of these  studies.   In  a  survey of  199  employees who worked  or  had  worked

02060                                VIII-14                         02/26/87

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with  toxaphene  between  1949  and  1977  with  exposures  ranging from 6 months to
26  years  (mean 5.23  yejrs),  20 employees  died,  one  with  cancer  of  the
colon.   None  of the  deaths  could be  attributed to  toxaphene  (IARC,  1979).
On  the  basis  of the available Information,  the  evidence  for  carclnogenldty
of  toxaphene 1n humans must be classified as Inconclusive.

    Several  Investigations  of the carcinogenic  potential  of  toxaphene  have
been  carried out  1n mice and rats.  While  Interpretations  of  the results of
these studies have  been  subject  to challenge,  the  conclusion  Is that  chronic
exposure  to  toxaphene can Induce hepatocellular  carcinoma  In  both mice  and
rats and shows some evidence of carcinogenic activity at  other sites as  well.

    The  most  thorough of  these  studies were  performed  by Tracor  JHco  Co.
under contract  to  the  NCI   (1979).   The  diets and  the  adjustments made In
toxaphene concentrations  In  the diets  have been described above  under  non-
carcinogenic  effects.   Treatment  was  discontinued  after  80  weeks and  the
studies   were  terminated  10-11 weeks  later  (mice)  or 28  weeks  later  (rats).
All animals  that  died  during the studies  and  all animals surviving at  the
termination of  the  studies were  submitted  for  pathologic  evaluation.  Of  the
male  rats, 90%  of  the high-dose, 94% of the low-dose and all  of the  control
group lived until at  least week  52 of the  study.  Of the female rats, 96% of
the high-dose,  92% of the low-dose  and all 10 controls  survived  beyond  the
52nd  week.   Although none  of  the  tumors   observed  1n  treated  animals  were
uncommon for the animal  strain  used,  certain tumors  and  hyperplastlc  tissues
were  present with  higher Incidence  1n  the treated  animals.   These Included
thyroid  folUcular  cell  adenomas and carcinomas,  and hyperplaslas.  Thyroid
folUcular cell adenomas and carcinomas combined  were  found  to  be elevated
02060                                VIII-15                         04/02/87

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statistically  In  both  male and female high-dose  groups  compared  with either
concurrent  or  h1stor1caT~controls from  the  same laboratory  (see  Tables  V-8
and  V-9).   In  the  female  rats  there was  also  a  statistically  significant
Increase 1n the cumulative  Incidence  of  tumors  of the pituitary (chromophobe
carcinomas  or  adenomas)  In  the  high-dose compared  with the  control  group.
The  authors  concluded  that  toxaphene administration was associated  with  an
Increase 1n  thyroid tumor  Incidence.  However,  It  should be  noted  that  the
adenomas Included 1n these evaluations are not classifiable as neoplasms.

    Following an  Independent  examination of  these hlstologlcal preparations,
Reuber  (1979)  also concluded that  toxaphene  administration  was  associated
with  an  Increase  In tumor  Incidence  even greater than  that  reported  by  NCI
(1979).  Since It 1s uncertain why  his  Interpretation of the tissues differs
from  the  original  NCI  Interpretation,   1t  Is  difficult to  draw  conclusions
from this study.

    In the  mice as  In  the rats,  survival  1n  treated groups  was high, 90-98%
of treated  and  control  animals surviving beyond  the  52nd  week of the study.
Of the  tumors  appearing  In  treated  animals,  none  were observed  In greater
Incidence compared  with controls with the  exception of those  In  the liver.
Hepatocellular  carcinomas  occurred  In treated mice  only,  with Incidences  of
98% and 69% In males  (see Table  V-10) at the high and low doses,  and 69% and
10%  1n  females (see  Table V-ll)  at  the high  and   low  doses,  respectively.
These  neoplasms  were  not  observed   In  control  animals of  either  sex,  but
hepatic nodules were  observed  In 20% of  the matched-control  males,  though
not  In  females.   On the basis of  these  findings, the authors concluded that
toxaphene caused  Increases  In  the  Incidence  of hepatocellular  carcinomas,


02060                                 VIII-16                         02/26/87

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and  hence,  was  carcinogenic  In  B6C3F1  mice.  Upon  examining the  tissues,
Reuber  (1979)  came  to a "qualitatively similar conclusion, although  again  he
reported a greater tumor Incidence than NCI (1979).

    In  addition  to  the previous study, a  number  of  other  much less  complete
studies  on  toxaphone  cardnogenlcHy  were conducted.   In a  study  conducted
by the  FDA  (Nelson,  1949), groups of  12  male and 12 female  rats assumed  to
be of  the  Osborne-Mendel strain were  fed  0,  25,  100,  400 or  1600 ppm  toxa-
phene  1n the  diet  for 107 weeks.   Further,  groups of  5  male and  5  female
rats were  fed 0,  40,  200 or  1000 ppm and  then  sacrificed  after 56  weeks.
Tissues were  sectioned only from  the  first set  of groups;  however,  an  Incom-
plete  histology  was  performed  on  other   groups.   Among  the second set  of
animal  groups  killed after 56  weeks,  only one neoplasm was  noted,  a  subcu-
taneous  neurosarcoma  In  a male  rat  given  1000  mg toxaphene/kg.  although
liver  hyperplasla  and hyper.plastlc  nodules  were  reported.    Thyroid  hyper-
plasla was also noted In treated animals,  especially In males.

    In  the  groups  used  In the  107-week  experiment, 2/3 female and  2/2  male
rats Ingesting 1600  mg toxaphene/kg  developed hepatic  carcinomas;  1/3  female
rats  1n each  of  the 400 and 1600  mg/kg   dose groups  developed  hyperplastk
nodules.  No  Incidence of  either  of  the above tumors was observed In control
or other treatment groups.

    Another  study  In mice demonstrated a  statistically  significant  Increase
In hepatic  neoplasms  (LHton  Blonetlcs,   Inc.,  1978).   Fifty-four  weanling
B6C3F1  mice  were  randomly  assigned  to  each of  four  groups.   Toxaphene
(X-16189-49),  as well  as the  dose  levels,  were  selected  and provided  by


02060                                VIII-17                         02/26/87

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Hercules  Inc.   The animals  were  administered toxaphene at  0,  7, 20  and  50
ppm  levels  In the  diet -for  18  months and  surviving  mice were  kept  on  the
control  diet  for  another  6  months.   All  survivors  were killed 24  months
after  the  Initiation  of  treatment.   Table  V-12  shows  the  effects   of
toxaphene on  survival.   The survival 1n both  treated  and  control groups  was
similar.

    The  hlstopathologlcal  evaluation  data  presented  In Table  V-13 shows  a
statistically significant  Incidence  of  hepatocellular  tumors (hepatocellular
carcinomas and hepatocellular adenomas) 1n male  mice fed  50  ppm toxaphene In
the diet as compared with  the controls.   However,  hepatocellular  tumor Inci-
dence was not statistically significant 1n females as compared with controls.

    Taken together,  the  results  of  these  studies  (LHton  Blonetlcs,  1978;
NCI, 1979) demonstrate that  toxaphene,  when  administered  chronically to mice
at  moderate- to   high-dose   rates,   Induces  hepatocellular  carcinomas   in
Osborne-Mendel rats and 1n  B6C3F1 mice, and  may  Induce tumors at other sites
as well.

    In another LHton  Blonetlcs  (1979) study, toxaphene was administered  to
50 weanling ARS  Golden Syrian hamsters of  each  sex and each dose at  levels
of  0,  100,  300  and 1000  ppm based on a  subchronlc  study.   The hamsters,
obtained  from the  Sprague-Oawley  laboratory  1n  Madison,  MI,  were  randomly
assigned  to  each  group.   The toxaphene  was mixed  In corn  oil  and blended
with  the food  that was  available  ad libitum.    The  animals  were  observed
during a  15-day  acclimation period  before Initiation  of the study and dally
02060                                VI11-18                         02/26/87

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afterward  for  general condition  and mortality  Treatment  was  continued for
18  months  for  the  females  and 21.5 months  for  the males  who  showed  a high
survival at 18 months.

    Animals  were  observed  until  spontaneous  death,   unless  moribund,  or
surviving  until  termination of  the experiment.   Moribund  animals  and those
surviving  until  termination were  sacrificed.   All  animals were  necropsled.
The  postmortem  observations   Included, a thorough  external  examination  and
collection of major  organs  and representative  tissues  from all  animals.  All
collected  tissues  from control and high-dose  animals,  and  target  organs and
gross  abnormalities   from  the  low- and  Intermediate-dose  animals,  were
evaluated  hlstopathologlcally.   Blood smears were  taken  from all  animals  at
sacrifice, but were  not  examined  unless  necessary  for  definition  of specific
lesions or diseases.   A  compound-related lowered body  weight was  observed  In
high-dose  males  until  12  months  of study.   Similar  effects were  noted  in
males  receiving  300 ppm from  months 2-7 and  for  a brief  period  In females
receiving  1000 ppm.   At  terminal  sacrifice, an  Increase  In liver  weight was
observed  In males  at  the  1000 ppm dose, which  correlated  to the  pathologic
finding of megahepatocytes 1n  6  of 21  males  observed.   A  decrease In heart
weight was noted  In males  at both  the 300  and  1000  ppm dose  levels.   A
decrease 1n the thyroid gland weights of the 1000 ppm females was  also noted.

    Only the liver  changes were supported  by  hlstopathology and  were there-
fore  Judged  to  be  treatment-related.   These  liver changes are  similar  to
those  accompanying   an  Induction  of liver  enzymes after  administration  of
chlorinated hydrocarbons and  other compounds.   Microscopic evaluation of the
tissues  did  not  reveal  any   Incidence  of  a  tumorlgenlc  effect  related  to


02060                                VI11-19                         02/26/87

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 toxaphene.   Even  when  the  Incidence  of  lymphoretlcular  neoplasms  of  all
 types  was  combined, no ^difference was  detected  between controls  and  dosed
 animals.  Under conditions of this study, toxaphene was not carcinogenic.

    It  Is  likely that the maximum tolerated dose was  not  employed In  these
 studies  (LHton  B1onet1cs,   Inc.,  1979).    In   the   subchronlc  study,  the
 maximum  tolerated  dose  estimate  was  based  on  evidence  of  liver  toxldty
 only.   No  hamster  at  any dose  level  died during  the study; no  changes  In
 general  appearance, condition  or  behavior  among  experimental  animals  was
 observed;  and  measurements  of  mean  body  weights  and  food  consumption
 Indicated no  compound-related  change.   In the chronic  study,  a  dose-related
 decrease  1n  body weight was observed,  but H was  temporary.  The  weight  of
 treated  animals  equaled  or surpassed  that  of  controls by  the end  of  1  year
 for all  treatment groups.  Treated animals  also  survived for a longer  period
 than  controls.   A  comparison   of  the  number  of  animals   In  each  treatment
group  surviving  to termination   of  the  experiment   using  an  X2  test  for
homogeneity  revealed  significant  differences  for both  males  (p<0.05)  and
 females  (p<0.001). '

Quantification of Carcinogenic Effects
    Since the  results  of two  bloassays  (NCI,  1979;  LHton B1onet1cs,  Inc.,
1978)  were  positive for  cancer Induction,  estimated risk  levels  for  toxa-
phene  In drinking  water  can  be  calculated using  a  linearized  multistage
model  as discussed  1n  the  appendices  to  the October  1980 Federal Register
notice   regarding   the  availability  of  Water  Quality  Criteria  Documents
 (Federal  Register,   1980).   Both  studies  are  considered  suitable for  the
02060                                VIII-20                         02/26/87

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derivation of  a  criterion.   The Litton 81onet1cs study was  selected  because
H allowed derivation of -a  slightly  more  conservative  criterion.   The animal
data used 1n the derivation are as follows:

          Dose             Incidence of Hepatocellular  Carcinoma  In Male  Mice
       (mq/kq/dav)         .	(No. Responding/No.  Tested)	
          0.0                                    10/53
          0.91                                   10/54
          2.6                                    12/53
          6.5                                    18/51
          Length of exposure =                   18  months
          Length of study »                      24  months
          Duration of Lifetime =                 24  months
          Body weight =                          0.030  kg
    With  these parameters,  the  carcinogenic potency  factor  for   humans  q  *
1s  1.131  (mg/kg/dayr1.    The  upper-limit  unit  risk  estimates  from  the
animal data are  derived  from a linearized multistage nonthreshold extrapola-
tion model  which 1s 'currently programmed as  GLOBAL  83.    Justification  for
Us  use   Is  presented  1n  EPA's  Guidelines  for  Carcinogen Risk  Assessment
(U.S. EPA,  1986b).   While recognizing that  alternative statistical modeling
approaches exist  [e.g.,  One-hH, Welbull, Log-ProbH,  and  LogH  models,  and
maximum  likelihood estimates],  the  range  of  risks described by using any of
these modelling  approaches  has  Uttle biological  significance   unless  data
can  be  used  to  support   the selection of  one model  over  another.   In  the
Interest  of  approach consistency and  of  providing  an upper bound estimate
for  the  potential  cancer  risk, the Agency recommends  the use of  the  linear-
ized multistage model.  EPA  considers  this model  and resulting risk estimate
to be an  upper limit value  1n the sense  that the true  risk  Is  unlikely to be
higher and  may be  lower  even zero.   An  established  procedure  does  not  yet
02060                                VIII-21                         12/07/87

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exist  for  making  "most likely" or  "best"  estimates  of  risk  within the range
of  uncertainty derived  by  the upper  and lower  limit  values.   In  order  to
derive a  drinking  water  concentration of  toxaphene  calculated  to  keep Indi-
vidual  risk below  10~5,  but  not   taking  Into  account  1ngest1on  of  seafood
or  Intake  of  toxaphene  by  other  sources  or  other  routes,   the  following
equation can be used.
              Hater  Concentration  =   10 '  x  70  kq	 =  0.31  vg/i
                                    1.131 mg/kg  x  2  I
where:
        10~5        = risk Level
        70 kg       = body weight of adult human
        1.131 mg/kg = carcinogenic potency factor (human q-)*)
        2 l         = dally water Intake for an average adult
    For  risk  levels  of  10~4,  10~s  and  10~6,  the corresponding  exposure
levels are 3.1, 0.31 and 0.031 vg/l, respectively.

    In summary,  toxaphene  has been shown to  Induce  cancer  1n two species of
animals,  while  evidence  for  cancer   Induction  1n  humans   Is  Inconclusive.
Toxaphene  1s,  therefore,  classified  as  an  animal carcinogen,  IARC  Category
2B  ranking,   meaning   that  toxaphene  1s  probably  carcinogenic   In  humans.
Applying  the  criteria  described  1n EPA's   guidelines  for  assessment  of
carcinogenic  risk  (U.S. EPA,  1986b),  toxaphene  may be classified  as  Group
B2:  probable  human  carcinogen,  meaning  there  1s  Inadequate  evidence  from
human studies and sufficient evidence from animal  studies.
02060                                VIII-22                         05/12/87

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Existing Guidelines. Recommendations and Standards
    Standards  for  toxaphene  1n air, water and  food  have  been established or
recommended  by  many  groups.   However,  most  of  these  standards  were  set
before the results  of  the  NCI  (1979)  or LHton Blonetlcs  (1978) bloassays of
toxaphene for  cardnogenldty  were  available.   On  May  25, 1977, the U.S. EPA
Issued a notice  of  rebuttable  presumption against  registration and continued
registration  of  pestlddal products containing  toxaphene (Federal  Register,
1977).   A  further  notice  was  Issued  1n  the  Federal  Register  (1982)  where
registration  for  use  under specified conditions was required after December
31,  1983 for  scabies  treatment of  beef cattle  and  sheep.   Depletion  of
existing  toxaphene stockpiles  was  allowed  until  December  31, 1986  under
limited  conditions  only.   Its sale and  distribution for  use as an Insecti-
cide  In  no-till  corn  and dry and southern  peas was  also  permitted  until
December 31,  1986 (Federal Register, 1982).
    The  ACGIH  (1977a)  established  a  TWA  value of  500 wg/m*  for  toxaphene
In the  air  of  the working  environment.   The ACGIH  (1977b)  based  this  stan-
dard  on unpublished  acute  and  chronic  toxlclty  studies  conducted  1n  the
1950s and on comparisons  of the toxlclty of  toxaphene  with DOT and llndane.
In  addition,   this  group   set  a  tentative  short-term exposure  limit  for
toxaphene of 1.0 mg/m* (ACGIH. 1977a).

    The  national  Interim  primary drinking water standard  for  toxaphene  Is  5
yg/l  (U.S.  EPA,  1976).  This  standard  1s  based  on  the  reported  organo-
leptlc  effects of  toxaphene  at concentrations  >5  yg/l   (Slgworth,  1965).
A  safe  level   of  25  w9/i was  also  calculated  based  on minimal  or  no
effects  1n rats after  they  were  fed toxaphene at  a concentration of 10 mg/kg


02060                                VIII-23                         04/06/87

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 In the diet, which was  estimated  to  give  an average dally dose of 1  mg/kg bw
 (Lehman, 1965).  This latter calculation used the following assumptions:

         weight of rat =                       300 g
         dally food consumption of rat =       50 g
         weight of average human adult =       70 kg
         average dally water Intake for man =  2 I
         safety factor =                       500
         dietary Intake =                      trace (assume zero)

    From these assumptions, the maximum safe  dally  dose  for  humans  was  esti-
mated  to  be 3.4  yg/kg/bw (U.S.  EPA,  1976).   It  should be noted,  however,
that the assumption  of  50 g dally food consumption  for  a  300  g  rat  1s  prob-
ably  excessively  high.   The  FDA  standard  of  5   yg/t for  bottled  water
(Federal Register, 1979)  1s  Identical  to  the U.S.  EPA's standard for drink-
Ing water.

    The National Ambient Mater Quality  Criterion  for the protection  of  human
health Is based upon  levels that  may  result In  Incremental  Increases In can-
cer  risk  over  the   lifetime.   For   risk  levels  of  10~5,  10'* and  10~7,
the  corresponding  criteria  are  7.1,  0.71  and  0.07   yg/l,  respectively
(U.S. EPA,  1980;  Federal Register,  1980).   If  the  above  estimates  are  made
for  consumption  of aquatic organisms  only, excluding consumption  of water,
the  levels  are 7.3,  0.73  and  0.07  wg/l,  respectively.  The criterion  was
derived  from  the  development  of hepatocellular  carcinomas  and neoplastk
nodules  In  B6C3F1  male  mice   given   several  doses  of  toxaphene  (Litton
B1onet1cs,  1978).
02060                                VIII-24                         04/02/87

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    The  NRC  (1977)  estimated  the RfO  of  toxaphene  for  man  at  1.25
This was  based  on  a study by FHzhugh  and Nelson  (1951),  In  which  rats  evi-
denced Increased liver weight and  hepatic  cell  enlargement  after  exposure  to
toxaphene at 25 ppm  In  the  diet  for  2 years.   In their estimation NRC  (1977)
assumed  the  dally  dose 1n  rats  during the FHzhugh  and  Nelson  (1951)  study
was equivalent  to  1.25 mg/kg/bw,  and  the  application of a safety  factor  of
1000 was  appropriate.   Then, assuming a human  body weight  of  70 kg and  a
dally  water  consumption  of 2 l,  NRC  (1977)  set  the  suggested NOAEL  from
water  at 8.75  yg/l  (assigning   20% of  the   total   RfO  to  water)  or  0.44
     (assigning IX of the total  RfO to water).
    The U.S.  EPA  criterion  for protection of freshwater  life  was  determined
to  be  0.013  yg/l as  a  24-hour  average,  and  the  concentration  should  not
exceed  1.6 vg/i  at  any time.   For  saltwater  aquatic  life  the  criterion
1s  0.019   vg/l   as   a  24-hour  average  with  a   maximum of   0.120  yg/l
(Federal Register, 1980;  U.S. EPA, 1980).

    The  International Joint  Commission  of  the  United  States  and  Canada
(1977)  has  recommended  a  water  standard  of 0.008  yg/l for  the  protection
of  aquatic  life.   This  standard  1s  based on the  study by Mayer  and  Mehrle
(1976)  In  which  they found that  toxaphene  at   0.039  yg/i caused  a  signif-
icant  Increase  1n mortality and  a  significant  decrease 1n growth  In brook
trout  fry  over a 90-day period.   The  standard of 0.008  yg/l  1s  obtained
by applying a safety  factor of 5.
02060                                VIII-25                         04/02/87

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Special Groups at Risk
    Sensitive  Suboooulatlon.   Children  may form  a  sensitive  subpopulatlon
for the toxic  effects  of  toxaphene.   Most  fatal  cases of toxaphene poisoning
have  Involved  children (Hayes,  1982).  It  Is possible  that  this  1s  due  to a
greater  Incidence of  high level  exposure  such  as  1ngest1on  of  toxaphene
solutions.   However,   there  Is  also  some  evidence  for  Increased  suscepti-
bility  1n  young  animals.   OffpsMng  of  rats fed only  50  wg/kg  bw/day,  from
3 weeks before mating  until  weaning  age,  showed delayed behavioral  develop-
ment  (Olson et al., 1980).

    Inadequate diet  may  result  1n   Increased  susceptibility to  toxaphene.
Central nervous  system effects  appeared earlier and  at lower concentrations
1n  rats  fed a protein deficient diet  (Boyd and Taylor,  1971).   Toxaphene-
Induced  Inhibition  of  enzymes  Involved  1n  carbohydrate  metabolism  was
greater In fasted than In fed chicks  (Srebocan et al., 1980a).

    Individuals with a less  Indudble xenoblotlc metabolizing  enzyme  system
may constitute a  sensitive subpopulatlon.  The  Importance  of the cytochrome
P-450  mediated detoxification  mechanism was Indicated  by the  2-  to  8-fold
Increase  1n  the  toxldty of Toxicant  B  1n mice  after treatment  with  the
cytochrome P-450  Inhibitor plperonyl  butoxlde (Saleh  et al.,  1977;  Turner et
al., 1977).

    Since  toxaphene  1s a  complex  mixture of mostly  chlorobornanes, differing
solubilities may  affect  the  actual   concentration  of the more  toxic  compo-
nents  1n drinking water.   The overall solubility of toxaphene Is quite low.
02060                                VIII-26                         04/02/87

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Brooks  (1974)  reported  a  mean   of  -3  mg/l,  while   Paris  et  al.  (1977)
reported  a  solubility of  0.5  mg/l at 25°C.   Almost all  of the  toxlclty  In
toxaphene 1s  due  to  Toxicants  A and  B (Hayes,  1982).   Toxicant  A, the larger
of  these  two fractions.  Is  less  polar  than the entire mixture  {Pollock  and
Kllgore,  1980)  Indicating  Its  solubllty  1n  water  1s  probably  less than toxa-
phene as  a  whole.   Toxicant B 1s  more polar and  thus  possibly  more  soluble,
but makes  up only -1% of  the  total  volume.  Thus,  1t   1s unlikely that  dif-
fering  solubilities  will  constitute  a major factor In  the  toxlclty  of toxa-
phene 1n water.  However,  since the  polar  fractions  are mutagenlc (Hooper  et
al., 1979)  the  more  water  soluble  components may  be  Important  1f mutagenesls
Is related to cancer.

    Toxaphene has  been detected  1n  drinking  water;  27/58  samples contained
toxaphene with  2  samples at  levels  >0.050 ppb (U.S.  EPA,  1977).  Toxaphene
In  drinking  water   has  not  been  detected In  some  environmental   surveys
(Schafer et al., 1969; Schulze et  al., 1973; Mattraw,  1975),  although It  has
been detected (up to  0.41  ppb)   1n  drinking  water after toxaphene  spraying
(Nicholson, 1964).  Toxaphene  In rain (up  to 0.5  ppb)  has also  been  detected
(Williams and Bldleman,  1978; Harder  et al.,  1980;  Munson,  1976;  Bldleman
and ChMstenson,  1979),  and this  exposure  route may be Important where  rain
water 1s utilized as drinking water.

    Interactions.   Induction of   hepatic  mlcrosomal  MFO  systems appears  to
account  for  mosy  of  the  Interactions   of  toxaphene  with  other  compounds.
In rats  pretreated  with  aldrln  or  dVeldrln and  evidencing  Increased liver
0-dealkylase  and 0-demethylase  activities,  toxaphene  96-hour  LD5Q  values
were  -2  times  higher  (Indicating decreased  toxldty)   than those   of  rats


02060                                VIII-27                         04/02/87

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given  no  pretreatment.    Similarly,   pretreatment  with  p.p'-DDT,  a  known
Inducer  of  hepatic  mlcrosomal  MFO,  resulted  1n  a 3-fold  Increase In  the
96-hour LD50 of toxaphene in rats (Delchmann and KepHnger,  1970).

    When  administered by  oral   Intubation  to  rats,  equltoxlc  combinations
(LD5Qs)  of  toxaphene with  parathlon,  dlazlnon or  trlthlon  were  less  toxic
than  would  be  expected,  based  on  the assumption  of  addHWe  toxldty  and
similar action (KepHnger and Delchmann, 1967).

    Toxaphene 1s commonly formulated with methyl parathlon, usually  In  a  2:1
mixture,  respectively,  for  control   of  certain   Insect  pests  of  cotton.
Chlordlmeform  Is  often  added   to  the  first  formulation  for  additional
control.  Combinations of  these  pesticides  were given to male  Swiss  mice by
gastric  Intubation;  25  mg  toxaphene/kg  bw  1n  0.16 ml  corn  oil;   12.5  mg
methyl parath1on/kg  *  25 mg  toxaphene/kg;  12.5 mg methyl parath1on/kg  <• 25
mg toxaphene/kg  *  3.25  mg  chlord1meform/kg (Crowder and WhUson,  1980).   In
these excretion-retention  studies, mortality  occurred only  In  groups receiv-
ing formulations containing methyl  parathlon  all  within  3 hours  of  dosing.
It appeared  that  methyl  parathlon toxldty  was slightly enhanced by chlor-
dlmeform  and  slightly  lowered  by  toxaphene  but   the  differences were  not
significant.

    The  possible  Interaction  of  methyl  parathlon and  toxaphene was  also
examined  on the  behavior   of  offspring  of  Sprague-Oawley rats  exposed  to
pesticides  on  days  7   through  15  of  pregnancy   (Crowder  et  al.,  1980).
Pregnant  rats  were given 1.0 mg/kg  bw methyl parathlon or 1.0  mg/kg methyl
parathlon  +  2.0  mg/kg  toxaphene  by  oral  gavage  1n  0.1  ml  corn  oil.

02060                                VIII-28                         04/02/87

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Toxaphen* alone  was  given from day 7  until  parturition  at  6 mg/kg/day.  Rat
pups  exposed In  utero  to these  doses of  the  pesticides alone  or  combined
demonstrated  few significant changes  1n  learning ability  as measured  by  a
simple  two-choice maze,  motor  skills or  behavior.   The   only  significant
change was  In  the additional time required  (4.3  days)  for  methyl parathlon-
toxaphene treated Utters to develop the righting reflex.

    Trlolo  et  al. (1982)  Investigated 1n  A/J  mice  the  effects  of  Ingested
toxaphene on  the Incidence  of lung tumors  Induced by  oral  administration of
benzo(a)pyrene  (BaP).   Groups  of 9-week-old  mice  (7-48/group)  were  fed  a
diet  containing  toxaphene (1n corn oil) at  100 ppm  for  12  weeks  or  200 ppm
diet  for  20 weeks.   Two  doses of  BaP (3  mg  each)  were given by  the oral
route:   the  first BaP dose  on  day 7  after  the Initiation  of  the toxaphene
diet, and  the  second dose on  day 21.  Nice were sacrificed after  12  or 20
weeks* and  prepared   for  tumor  analysis  and   analysis   of   BaP  hydroxylase
activity.  Mice  that  received  toxaphene 1n the diet alone,  or  toxaphene and
BaP,  showed an  Increase 1n  BaP  hydroxylase  activity  In   the  liver  and  a
decrease 1n enzyme activity  1n  the  lung.   Inhibition of  lung BaP hydroxylase
activity was  paralleled   by  a  reduction  In  BaP-1nduced  lung tumors  In mice
fed toxaphene.

    Over a period of  60  days  Bogachuk and Fllenko (1978) studied the effects
of  vertical  vibration,  toxaphene,  and a combination  of these  physical  and
chemical  factors In  80  Immature  Inbred male  rats  of  the Hlstar  and August
strains.  Rats  were  divided Into  four groups  consisting of  10  rats  of each
strain;  group  1, control; group  2,  vertical vibration  (frequency  50 hertz,
amplitude  1.25  mm,  30  m1n/day);  group  3,  0.01 L05Q   toxaphene  perorally


02060                                VI11-29                         04/02/87

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 (L050  not given);  group  4,  combined  action  of  vibration and  toxaphene  at
 the  same  levels as  In  groups  2 and 3.  Experiments were  conducted  on  every
 group at  the  same time each day for 60  days  and  covered  the  period  of  rapid
 growth of  the  animals.  Decreased  body weight, kidney  weight,  volume,  linear
 dimensions and thickness  of  the cortex were  observed 1n  the  three treated
 groups.   The  changes   were  most  pronounced  1n  the  group  exposed to  the
 combination of  toxaphene  and  vibration; the combined  effects  appeared  to  be
 additive.

    Mount  et al.  (1980) reported an outbreak  of toxaphene poisoning  1n  10  of
 15 crossbred sows  1n mldgestatlon.  The animals  ate feed  that was  poured  on
 the  wet  floor   of  a holding  pen  treated a  month earlier  with  a  commercial
 spray containing 45X toxaphene  and  2%  Undane.  Within 30 minutes,  10  of the
animals developed  an acute nervous  disorder.   Two  sows died  within 2  hours
 from  the  onset  of  clinical   signs.    In   the  others,   severity  of   signs
diminished without  treatment  after 2.5  hours.  Toxaphene  and  Undane  levels
 (mg/kg wet weight)  found  1n the tissue  were:   cerebrum,  2.0  and 0.02;  cere-
bellum, 4.0  and 0.03;  and  serum  (hemolyzed),  0.3  and 0.002,  respectively.
Although  the  levels of Undane  1n the  tissues were below levels  associated
with clinical  Intoxication,  the presence of Undane may  have  contributed  to
the severity of the  signs.

    Starvation  may  cause  greater mortality to  toxaphene  than  optimal  diets.
Thus,  rats fed a  protein-deficient diet  showed acute  oral  LD.g values  of
80+19 mg/kg  bw, whereas  for  animals  fed  standard  laboratory  chow  the LD5Q
was  220*33 mg/kg.   The LD5Q  of rats  fed a  high protein  diet  was  293*31  mg
 toxaphene/kg  bw,   Indicating  a  protective  effect  of  dietary  protein.   CNS

 02060                                VIII-30   •                     04/02/87

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effects  appeared  earlier and  at  lower concentrations  than  In control  rats
(Boyd and  Taylor,  1971).  Toxaphene (5 or 10  ppm In  the diet),  administered
to chicks  after 24  hours without  food,  caused  lower  activities of glucose-6-
phosphatase,  pyruvate  carboxylase  and  phosphoenolpyruvate carboxyklnase  1n
1-day-old  N1ck  chick   cockerels   than  did  toxaphene   or  starvation  alone
(Srebocan  et  al., 1980a).   Heat stress  In  poultry did not appear  to Interact
significantly with  toxaphene  (Srebocan  et  al.,  19806).   Based  on  the absence
of toxaphene-lnduced changes  1n plasma  levels  of  pyruvlc and  lactic adds  In
unstressed  rats,  Peakall  (1979)  Implied  that  rats   had  to  be "stressed  In
some way"  to  obtain depressed blood LDH activity  as  claimed  by  Kuz'mlnskaya
and Alekhlna  (1976) and Gertlg and Nowaczyk (1975).

    The  toxldty  of Toxicant B to  mice Is  Increased  by a factor  of  2-8  by
administration  of  plperonyl  butoxlde  (Saleh  et  al.,  1977;  Turner et  al.,
1977), Indicative of the Importance of  cytochrome P-450 mediated  detoxifica-
tion mechanisms.   This  would  Indicate  that any  chemical that depressed  or
enhanced  mlcrosomal function  might affect  the  ultimate toxldty  to  toxa-
phene.   It has  been  postulated  that  1n  young  white  Leghorn chickens  the
detoxification  systems  are  not  yet  fully  Induced,  making younger  animals
more susceptible than  older  ones  (Bush  et  al.,  1977).  Thus,  age may also  be
an Interacting  variable,  a  fact also noted  during behavioral  testing  (Olson
et al., 1980).
02060                                VIII-31                         04/02/87

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                                IX.   REFERENCES

ACGIH  (American  Conference of  Governmental  Industrial Hyglenlsts).   1977a.
TLVs: -Threshold  Limit  Values  for  Chemical Substances and Physical  Agents  1n
the Workroom Environment with Intended Changes  for 1977.   Cincinnati,  OH.

ACGIH  (American  Conference of  Governmental  Industrial Hyglenlsts).   1977b.
Documentation of Threshold Limit Values, 3rd ed.   Cincinnati,  OH.

Alekhlna,  S.M.   and  U.  A.  Kuz'mlnskaya.   1980.   Isoenzymatlc  spectrum  of
lactate  dehydrogenase  1n rat  Hver  and  myocardium tissues  as  affected  by
polychlorocamphene.  Ukr. B1okh1m. Zh.  52:483-485.

Allen,  A.L.,  L.D.  Koller,  and  G.A.  Pollock.   1983.    Effect  of  toxaphene
exposure  on  Immune  responses  In mice.   J. Toxlcol.  Environ.  Health.   11:
61-69.

Aller,  H.E.,  and C.D.  Hansen.   1981.  Safened  phosphorothlolate  pestlddal
compositions.  U.S. Patent 4251523.

Anonymous.  1982.  Herblcldal mixtures.  Res. Disci.  219: 253-254.

Archer,  T.E.  and  O.G.  Crosby.   1966.   Gas  chromatographlc  measurement  of
toxaphene  In milk,  fat, blood,  and  alfalfa  hay.  Bull.  Environ.  Contain.
Toxlcol.  1:  70-75.
02070                               IX-1                             02/26/87

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Arscott,  G.H.,  S. Mlchener  and  O.D. Bills.   1976.   Effect of  toxaphene  on
the performance of wh1te_leghorn  layers  and  their  progeny.   Poult.  Scl.   55:
1130-1131.

Badaeva.  L.N.   1979.   Effect of  some pesticides on  chollnesterase  activity
In the cardiac neural elements of  pregnant animals and  their  fetuses.  Arkh.
Anat. Glstol. EmbMol.  76(4): 68-71.

Badaeva,  L.N.   1981.   Experimental study of the postnatal  neurotoxlc  effect
of chloroorganlc pesticides.  Folia Morphol.   29: 113-114

Barbehenn, K.R.  and  H.L.  ReUhel.   1981.   Organochlorlne  concentrations  1n
bald eagles:  Brain/body  I1p1d relations  and hazard evaluation.   J.  Toxlcol.
Environ. Health.  8:  325-330.

Barthel,  E.   1976.   High  Incidence  of   lung  cancer  1n  persons  with  chronic
professional   exposure to  pesticides In agriculture.  Z.  Erkr.  Assn.-dry.
146:  266-274.  (Cited 1n IARC, 1979)

Barthel,  E.   1981.   Increased risk of lung cancer on pesticide-exposed  male
agricultural  workers.   J. Toxlcol. Environ.  Health.   8:  1027-1040.

Bateman, G.Q., C. Blddulph,  J.R.  Harris, et  al.   1953.  Transmission  studies
of milk  of dairy cows fed  toxaphene-treated hay.   J. Agrk. Food  Chem.   1:
322-334.
02070                               IX-2                             02/26/87

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Baumler,  W.   1975.   Nebenwerkungen  von Toxaphene  auf  Mause.   Anz.  Schaed-
llngsk. Pflanzenschutz. Umweltschutz.  48:  65-71.

Bldleman, T.F.  and E.J.  Chrlstensen.   1979.   Atmospheric removal  processes
for h1gh-molecular-we1g*it organochloMnes.   J.  Geophys.  Res.  84:  7857-7862.

Bogachuk, G.P.  and V.E. Fllenko.  1978.   Effect  of general  vertical  vibra-
tion  and  polychlorocamphene  on the  kidneys  of  Inbred  rats.   Arkh.  Anat.
Glstol. EmbMol.  75(7): 24-27.

Boots  Hercules  Agrochemlcals,  Inc.   n.d.  Boots Hercules Toxaphene  Insecti-
cide Summary of Tox1colog1cal Investigations.   Bulletin  T-1050.

Boshoff, P.R. and  V.  PretoMus.  1979.  Determination of toxaphene  In  milk,
     *
butter and meat.  Bull. Environ. Contam. Toxlcol.   22:  405-412.

Boyd,  E.M.  and  F.I.  Taylor.  1971.  Toxaphene  toxldty  In protein-deficient
rats.  Toxlcol.  Appl.  Pharmacol.  18: 158-167.

Braun,  R.K.,  F.C. Neal  and H.A.  Nelson.   1980.   Acute death and  abnormal
behavior In beef cattle  resembling toxaphene poisoning.   Fla.  Vet.  J.   9(3):
15-16.

Brooks,  G.T.    1974.   Polychloroterpene  Insecticides  {toxaphene).    JJK
Chlorinated  Insecticides,  Vol. 1, Technology  and  Application.  CRC  Press.
p. 205-210.
02070                               IX-3                             02/26/87

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Buck, W.B., G.D.  Osweller and  G.A.  Van  Gelder.   1976.   Insecticide  levels  In
tissues  associated  with _tox1c1ty:   a literature review.  Vet.  Hum.  Toxlcol.
23: 34-42.  (Cited 1n Mount and Oehme, 1981)

Bush, P.B.,  3.T.  K1ker,  R.K.  Page,  et  al.  1977.   Effects of  graded  levels
of  toxaphene on poultry residue  accumulation,  egg  production,  shell  quality,
and hatchabllUy 1n white leghorns.   J.  AgMc.  Food Chem.   25:  928-932.

Bush, P.B.,  M.  Tanner, J.T. K1ker  et al.   1978.  Tissue residue studies  on
toxaphene 1n broiler chickens.   J.  Agrlc.  Food  Chem.   26:  126-130.

Cabral,   J.R.P.,  f.  Raltano, T.  Mollner,   S. Bronczyk  and P.  Shublk.   1979.
Acute toxlclty  of  pesticides  1n hamsters.  Toxlcol.  Appl. Pharmacol.   48:
A192.

Cairns,  T., E.G. Slegmund and  J.E.  Froberg.  1981.   Chemical  'lonlzatlon  mass
spectrometrlc examination  of metabolized  toxaphene  from milk  fat.   Blomed.
Mass Spectrom.   8: 569-574.

Causey,   K.,  S.C.  Mclntyre,  Jr., and  R.W. Rlchburg.   1972.   OrganochloMne
Insecticide residues  1n quail,  rabbits, and deer from  selected  Alabama  soy-
bean fields.  J. Agrlc. Food Chem.   20:  1205-1209.

Chandurkar, P.S.   1977.   Metabolism of toxaphene components 1n  rats.   Ph.D.
Thesis,  Univ. Wisconsin,  Madison, HI.
02070                               IX-4                             02/26/87

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Chandurkar,  P.S.  and  F.  Matsumura.   1979a.  Matabollsm  of  toxaphene  com-
ponents 1n rats.  Arch. Environ. Contam.  Toxlcol.   8:  1-24.

Chandurkar,  P.S.  and  F.  Hatsumura.   1979b.  Metabolism  of  toxicant  C  of
toxaphene 1n rats.  Bull.  Environ. Contam. Toxlcol.   21:  539-547.

Chandurkar, P.S., F. Matsumura  and T.  Ikeda.   1978.   Identification  and  tox-
1c1ty  of  toxicant  Ac, a  toxic  component  of  toxaphene.   Chemosphere.    7:
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02070                               IX-8                             02/26/87

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02070                               IX-15                            02/26/87

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02070                               IX-19                            02/26/87

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02070                               IX-20                            11/30/87

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02070                               IX-21                            02/26/87

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Peakall,  O.B.   1979.   Ef_fect  of toxaphene on pyruvlc and  lactic  add  levels
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02070                               IX-22                            02/26/87

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Pollock,  R.H.   1958.   Toxaphene-llndane  poisoning  by cutaneous  absorption.
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02070                               IX-23                            02/26/87

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Saleh,  M.A.,  u.V.  Turner and  J.E.  Caslda.   1977.   Polychlorobornane com-
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02070                               IX-24                            02/26/87

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02070                               IX-26                            02/26/87

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02070                               IX-27                           02/26/87

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02070                               IX-28                            02/26/87

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02070                               IX-29                            02/26/87

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02070                               IX-30                            02/26/87

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