xvEPA
               United States
               Environmental Protection
               Agency
               Office of Water
               Regulations and Standards
               Criteria and Standards Division
               Washington DC 20460
EPA 440 5-80-062
October 1980
Ambient
Water Quality
Criteria  for
Diphenylhydrazine

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      AMBIENT WATER QUALITY CRITERIA FOR

             DIPHENYLHYDRAZINE
                 Prepared By
    U.S.  ENVIRONMENTAL PROTECTION AGENCY

  Office of Water Regulations and Standards
       Criteria and Standards Division
              Washington, D.C.

    Office of Research and Development
Environmental Criteria and Assessment Office
              Cincinnati, Ohio

        Carcinogen Assessment Group
             Washington, D.C.

    Environmental Research Laboratories
             Corvalis, Oregon
             Duluth, Minnesota
           Gulf Breeze, Florida
        Narragansett, Rhode Island

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                              DISCLAIMER
     This  report  has been reviewed by  the  Environmental  Criteria and
Assessment Office,  U.S.  Environmental  Protection Agency,  and approved
for publication.  Mention of trade names or commercial products does not
constitute endorsement or recommendation for use.
                          AVAILABILITY  NOTICE
      This  document is available  to  the public  through  the National
Technical Information Service, (NTIS), Springfield, Virginia  22161.
                                   11

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                               FOREWORD

    Section 304  (a)(l)  of  the Clean Water Act  of  1977  (P.L.  95-217),
requires the Administrator  of the Environmental Protection Agency  to
publish criteria  for water  quality  accurately reflecting  the  latest
scientific knowledge on the kind and extent of all identifiable effects
on  health  and  welfare which  may be  expected from  the presence  of
pollutants in any body of water, including ground water.  Proposed water
quality criteria  for the 65  toxic  pollutants  listed  under section 307
(a)(l) of  the  Clean Water  Act  were  developed  and  a notice  of  their
availability was published for public  comment on March 15, 1979 (44  FR
15926), July 25,  1979 (44 FR  43660), and October 1,  1979  (44 FR 56628).
This document  is  a revision  of  those  proposed criteria  based  upon  a
consideration of  comments received from  other  Federal Agencies,  State
agencies,   special  interest  groups,  and  individual  scientists.    The
criteria contained in this document replace any previously published EPA
criteria for  the  65 pollutants.   This criterion  document  is  also
published  in satisifaction of paragraph 11 of the Settlement Agreement
in  Natural  Resources  Defense Council, et   al.  vs. Train,  8  ERC 2120
(D.D.C. 1976),  modified, 12 ERC 1833 (D.D.C.  1979).

    The term "water  quality criteria"  is used  in  two sections  of the
Clean Water Act, section 304 (a)(l)  and section 303 (c)(2).  The term has
a different program  impact  in  each section.   In section 304,  the term
represents a non-regulatory,  scientific  assessment of  ecological  ef-
fects. The criteria  presented  in  this  publication  are such scientific
assessments.   Such  water  quality criteria  associated  with  specific
stream uses when  adopted as  State water quality  standards under section
303  become  enforceable maximum acceptable  levels  of  a  pollutant  in
ambient waters.  The water quality criteria adopted in the State water
quality standards could have the same  numerical limits as the  criteria
developed  under section  304.  However, in many situations  States may want
to adjust  water quality criteria developed under section 304 to reflect
local  environmental conditions  and  human  exposure  patterns  before
incorporation  into  water quality  standards.    It  is not  until  their
adoption as part of the State water quality standards that the criteria
become regulatory.

    Guidelines  to assist the  States  in  the modification  of  criteria
presented   in   this  document,  in  the  development of  water  quality
standards,  and  in  other water-related programs of this Agency,  are being
developed  by EPA.
                                    STEVEN SCHATZOW
                                    Deputy Assistant Administrator
                                    Office of Water Regulations and Standards
                                   111

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                                ACKNOWLEDGEMENTS
Aquatic Life Toxicology:

    William A. Brungs, ERL-Narragansett
    U.S. Environmental Protection Agency
John H. Gentile, ERL-Narragansett
U.S. Environmental Protection Agency
Mammalian Toxicology and Human Health Effects:

    Joseph Borzelleca (author)
    Medical College of Virginia

    Richard Carchman (author)
    Medical College of Virginia

    Terence M. Grady (doc. mgr.) ECAO-Cin
    U.S. Environmental Protection Agency

    Bonnie Smith (doc. mgr.) ECAO-Cin
    U.S. Environmental Protection Agency

    Edward Calabrese
    University of Massachusetts

    Thomas Clarkson
    University of Rochester

    Joan Cranmer
    University of Arkansas

    Patrick Durkin
    Syracuse Research Corporation

    George C. Fuller
    University of Rhode Island

    Roy E. Albert, CAG*
    U.S. Environmental Protection Agency
Si Duk Lee, ECAO-Cin
U.S. Environmental Protection Agency

Krystyne Locke, HED
U.S. Environmental Protection Agency

Steven D. Lutkenhoff, ECAO-Cin
U.S. Environmental Protection Agency

Herbert Schumacher
National Center for Toxicological Research

Samuel Shibko
U.S. Food and Drug Administration

Carl C. Smith
University of Cincinnati

Jerry F. Stara, ECAO-Cin
U.S. Environmental Protection Agency

S.L. Schwartz
Georgetown University

Norman Trieff
University of Texas Medical Branch
Technical Support Services Staff:  D.J. Reisman, M.A. Garlough, B.L. Zwayer,
P.A. Daunt, K.S. Edwards, T.A. Scandura, A.T. Pressley, C.A. Cooper,
M.M. Denessen.

Clerical Staff:  C.A. Haynes, S.J. Faehr, L.A. Wade, D. Jones, B.J. Bordicks,
B.J. Quesnell, T. Highland, B. Gardiner


*CAG Participating Members:  Elizabeth L. Anderson, Larry Anderson, Ralph Arnicar,
Steven Bayard, David L. Bayliss, Chao W. Chen, John R. Fowle III, Bernard Haberman,
Charalingayya Hiremath, Chang S. Lao, Robert McGaughy, Jeffrey Rosenblatt,
Dharm V. Singh, and Todd W. Thorslund.
                                       IV

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                                   TABLE OF CONTENTS



                                                                     Page

Criteria Summary

Introduction                                                         A~l

Aquatic Life Toxicology                                              B-l
    Introduction                                                     B-l
    Effects                                                          B-l
         Acute Toxicity                                              B~|
         Summary                                                     B'J
    Criteria                                                         B'l
    References                                                       B~3

Mammalian Toxicology and Human Health Effects                        C-l
    Exposure                                                         C-l
         Ingestion from Water                                        C-l
         Ingestion from Food                                         C-l
         Inhalation, Dermal, and Other Sources                       C-2
    Pharmacokinetics                                                 C-3
         Absorption, Excretion, and Distribution                     C-3
         Metabolism                                                  C-3
    Effects                                                          C-3
         Acute, Subacute, and Chronic Toxicity                       C-3
         Synergism and/or Antagonism                                 C-5
         Teratogenicity                                              C-5
         Mutagenicity                                                C-5
         Carcinogenicity                                             C-6
    Criterion Formulation                                            C-12
         Existing Guidelines                                         C-12
         Current Levels of  Exposure                                  C-12
         Special Groups at  Risk                                      C-12
         Basis  and Derivation of Criterion                           C-12
    References                                                       C-16
Appendix                                                             C-19

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                              CRITERIA DOCUMENT
                              DIPHENYLHYORAZINE
CRITERIA
                                 Aquatic  Life
    The available data  for  1,2-diphenylhydrazine  indicate that acute  toxic-
ity to  freshwater aquatic life  occurs at concentrations as low as  270  pg/1
and would occur  at  lower concentrations   among  species  that are more  sensi-
tive than those  tested.   No  data are  available  concerning the  chronic  toxic-
ity of 1,2-diphenylhadrazine to sensitive freshwater aauatic life.
    No  saltwater organisms  have  been  tested with  1,2-diphenylhydrazine  and
no statement can be made concerning acute or chronic toxicity.

                                 Human Health
    For the  maximum protection  of  human  health from the  potential  carcino-
genic effects due to  exposure of diphenylhydrazine  through ingestion of  con-
taminated water  and contaminated aauatic  organisms,  the ambient water  con-
centrations  should  be  zero  based on the non-threshold assumption  for  this
chemical.  However,  zero level   may  not  be  attainable  at the  present  time.
Therefore,  the   levels  which may  result  in  incremental  increase of  cancer
risk  over  the  lifetime  are   estimated  at  10"5,  10  ,  and  10~ .   The
corresponding recommended criteria are 422  ng/1,  42 ng/1,  and 4 ng/1,  re-
spectively.   If  the above estimates are made for consumption  of  aquatic or-
ganisms only, excluding  consumption  of water, the levels  are  5.6  ug/1,  0.56
ug/1, and 0.056  ug/1, respectively.
                                      VI

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                                 INTRODUCTION

    Diphenylhydrazine  exists  as an  asymmetrical  isomer,  1,1-diphenylhydra-
zine, and  a symmetrical  isomer, 1,2-diphenylhydrazine (hydrazobenzene).   The
hydrochloride of  1,1-diphenylhydrazine  is  used as a reagent for  the  sugars,
arabinose  and  lactose  (Windholz,  1976).   1,2-Diphenylhydrazine  is used  in
the synthesis of  phenylbutazone (Wenner, 1967) and as the  starting material
in the  manufacture of benzidine,  an  intermediate in the production of  dyes
(Lurie, 1964).
    The primary method of commercial  production of this compound is  the re-
                                                    +3       +2
duction  of  nitrobenzene  by  catalysts  such  as  Fe    or   Zn     in  alkaline
solution.  8y this procedure, nearly quantitative yields are obtained  (Kirk-
Othmer, 1963).  Figure 1 depicts this process as well  as  the  by-products of
diphenylhydrazine, azobenzene and azoxybenzene.
    In  1977  the commercial  production of 1,2-diphenylhydrazine was in  excess
of  1,000   Ibs.  [Stanford  Research Institute  (SRI),  1977].   However,  this
figure  is  probably an underestimate of  the  amount  of diphenylhydrazine  that
was actually available.   Diphenylhydrazine is produced  in  several  synthetic
processes  as an  intermediate  or as a contaminant,  but  it  is not  possible to
estimate these quantities, which are probably substantial.
    The reaction  of  1,2-diphenylhydrazine  with acid results in the benzidine
rearrangement (Kenner,  1968).   This reaction  is  presented  in  Figure  2.   In
addition to benzidine,  other products  formed include  diphenyline, o-benzi-
dine, and  o-semidine.  In  the stomach,  1,2-diphenylhydrazine can  be convert-
ed  into benzidine,  a  known  human  carcinogen  [Haley,  1975;  International
Agency for Research on Cancer (IARC),  19721.
                                      A-l

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       N-N
*Azobenzene
                                                Nitrobenzene
                          Alka^ne
                            Zn
*Azoxybanzene
                                                  1,2-diphenylhydrazine
                                FIGURE  1

                  Synthesis  of 1,  2-diphenylhydrazine

                        Source:  Williams, 1959


                  *Carcinogenic
                                  A-2

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                                H
     2-diphenylhydrazine
* Carcinogenic
                                                                     *Benzidine
                                                        Diphenyline
                                     o-Benzidine
                          o-Semidine





                                              FIGURE 2



                          Benzidine Rearrangement of 1,  2-diphenylhydrazine




                                       Source:   Williams,  1959
                                               A-3

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    The  structure  and physical data  for  1,2-diphenylhydrazine  are presented
in Figure 3.
    No data  were found on  the environmental  presence or  persistence  of di-
phenylhydrazines, except for  one  report of detection in drinking  water  at a
concentration of 1  ug/1  (U.S.  EPA,  1975).  1,1-Diphenylhydrazine and 1,2-di-
phenylhydrazine have  been characterized as  slightly  soluble  and insoluble in
water, respectively (Windholz, 1976;  Bennett,  1974).  No quantitative  data
were  found  for  the water  solubilities  and  vapor  pressures  of  these  com-
pounds;  consequently,  no  predictions  can  be made about their  persistence in
water.
                                      A-4

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                          FIGURE 3
 1,2-diphenylhydrazine:  Chemical and Physical Properties
                    Source:   Weast,  1978.
Synonyms
- Hydrazobenzene
  Symmetrical diphenylhydrazine
  N, N'-Diphenylhydrazine
  N, N'-Bianiline
  1,1' -Hydrazodibenzene
Cas No.
  530-50-7
  Molecular weight = 184.24
  Melting Point = 131°C
  Boiling Point = 220°C
Solubility = Slightly soluble in water
             Very soluble in benzene, ether, alcohol
                            A-5

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                                  REFERENCES

Bennett, H. (ed.)   1974.   Concise Chemical  and  Technical  Dictionary.   Chemi-
cal Publishing Co.  Inc., New York.

Haley,  T.J.   1975.   Benzidine revisited:  A review  of  the literature  and
problems associated with the use of benzidine.

International  Agency for Research on  Cancer.  1972.   Aromatic  amines.   Mono-
graph on the evaluation of carcinogenic risk of  chemicals to man.   1:  69.

Kenner, J.  1968.  Benzidine rearrangement.  Nature.   219: 153.

Kirk,  R.E.  and  D.F.  Othmer.   1963.   Encyclopedia of  Chemical  Technology.
2nd ed.  John Wiley and Sons, Inc., New York.

Lurie,  A.P.   1964.   Benzidine.   In:  Kirk-Othmer Encyclopedia  of  Chemical
Technology.  2nd ed.  Interscience Publishers, New York.  3: 408.

Stanford Research  Institute.  1977.   Directory  of  Chemical  Producers.   Menlo
Park, California.

U.S.  EPA.   1975.   Preliminary assessment of  suspected  carcinogens  in  drink-
ing water.  Off. Tox. Subst., Washington, D.C.

Weast,  R.C.   (ed.)   1978.   Handbook   of Chemistry  and  Physics.   59th  ed.
Chemical Rubber Company Press, West Palm Beach,  Florida.
                                     A-6

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Wenner, W.   1967.  Malonlc  Acid and Derivatives.   In;  Kirk-Othmer Encyclo-
pedia of  Chemical Technology.   2nd  ed.   Interscience Publishers,  New York.
12: 857.

Williams, R.   1959.   Detoxication  Mechanisms.   John  Wiley and  Sons,  Inc.,
New York.  p. 480.

Windholz, M. (ed.)  1976.  The  Merck Index.   Merck and Co. Inc., Rahway, New
Jersey.
                                       A-7

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Aquatic Life Toxicology*
                                 INTRODUCTION
    Toxicity tests  with 1,2-diphenylhydrazine and  the  bluegill and  Daphnia
magna have  been conducted  using  static procedures, and  the results demon-
strated adverse  effects as low as  270  ug/1.   No data are available for  any
saltwater species.
                                    EFFECTS
Acute Toxicity
    The  48-hour  EC™  for Daphnia magna  and  the  96-hour  LCj.g  for  the
bluegill are 4,100  ug/1 and 270  ug/1,  respectively,  for  1,2-diphenylhydra-
zine (Table 1).
Summary
    Only  two  freshwater  species have  been  tested with  1,2-diphenylhydra-
zine.  The  range of 50 percent  effect  levels for  the  bluegill and  Daphnia
magna was 270  to  4,100  ug/1. No  data  are  available  for  1,2-diphenylhydrazine
and saltwater organisms.
                                   CRITERIA
    The available data for  1,2-diphenylhydrazine indicate that  acute  toxici-
ty  to  freshwater aquatic  life  occurs at concentrations  as  low as 270 ug/1
and would occur  at  lower concentrations among species  that  are more sensi-
tive than those tested.  No data  are  available concerning  the  chronic toxic-
ity of 1,2-diphenylhydrazine to sensitive  freshwater aguatic  life.
    No saltwater  organisms have  been  tested with  1,2-diphenylhydrazine  and
no statement can be made concerning  acute  or chronic toxicity.
*The reader is referred to the  Guidelines  for  Deriving  Water  Quality Criter-
ia for the Protection of Aquatic Life and  Its Uses in order to  better under-
stand  the following  discussion  and  recommendation.   The  following  tables
contain the appropriate data  that were found in the literature,  and  at  the
bottom of each table  are  calculations for deriving  various measures  of  tox-
icity as described in the  guidelines.
                                      B-l

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Table I.  Acut* valiM* for 1,2-dlpb*oylhy
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                                  REFERENCES
U.S. EPA.   1978.   In-depth  studies  on health  and  environmental  impacts  of
selected  water  pollutants.   U.S.  Environ.   Prot.  Agency,   Contract   No.
68-01-4646.
                                      B-3

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Mammalian Toxicology and Human Health Effects
                             EXPOSURE
Inqestion from Water
     1,2-Diphenylhydrazine  (DPH)  was not detected in the finished
water of any of 10 cities selected for a  detailed  study by U.S. EPA
(1975).  However,  the  same study demonstrated the presence of DPH
in drinking water in concentrations  up to 1 yg/1  (1 ppb).
Ingestion from Food
     There  are  no available  data identifying DPH as  a direct or
indirect food additive, or as a naturally occurring constituent of
any food.
     A bioconcentration factor (BCF)  relates the concentration of a
chemical in aquatic  animals to the  concentration  in  the water in
which they  live.   The steady-state  BCFs for  a lipid-soluble com-
pound in the tissues of various aquatic  animals seem to  be propor-
tional to the  percent  lipid in the  tissue.   Thus,  the per capita
ingestion of a  lipid-soluble chemical can be estimated  from the per
capita consumption of fish and shellfish, the  weighted  average per-
cent lipids of consumed fish and  shellfish, and a steady-state BCF
for the chemical.
     Data from a recent survey on fish and shellfish consumption in
the United  States were  analyzed  by SRI  International  (U.S.  EPA,
1980).  These data were  used to  estimate that the per capita con-
sumption of  freshwater  and  estuarine fish  and  shellfish  in the
United States  is 6.5  g/day  (Stephan,  1980).   In addition,   these
data were used  with data on the fat content of  the  edible  portion of
                               C-l

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the  same  species  to estimate  that  the weighted  average percent
lipids for consumed freshwater and estuarine fish and shellfish is
3.0 percent.
     No measured  steady-state  bioconcentration  factor  (BCF)  is
available for  lf2-diphenylhydrazine,  but the  equation  "Log BCF =
(0.85 Log P) - 0.70" can  be  used  (Veith, et al. 1979) to estimate
the BCF for  aquatic organisms that contain about 7.6 percent lipids
(Veith, 1980)  from  the  octanol/water partition  coefficient  (P) .
Based on a measured log P  value  of 2.94  (Hansch and Leo, 1979), the
steady-state bioconcentration  factor  for 1,2-diphenylhydrazine is
estimated to be 63.  An adjustment factor of 3.0/7.6 » 0.395 can be
used to adjust the  estimated  BCF  from  the 7.6 percent lipids on
which the equation  is  based  to  the  3.0  percent lipids that is the
weighted  average  for  consumed  fish  and   shellfish.   Thus,  the
weighted average bioconcentration factor for 1,2-diphenylhydrazine
and the edible portion of all aquatic organisms consumed by Ameri-
cans is calculated to be 63 x 0.395 = 24.9.
Inhalation, Dermal, and Other Sources
     Laboratory workers, workers in forensic medicine, and workers
involved in  the  manufacture  of dyes,  certain Pharmaceuticals, and
chemicals, are  subject to occupational  exposure  to 1,2-diphenyl-
hydrazine.  Both inhalation and dermal contact  are possible routes
of exposure  in these settings.  However, no experimental data are
available to quantitate  either the dermal  or   inhalation exposure
risks to this population.
                               C-2

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                         PHABMACOKINETICS
Absorption, Excretion, and Distribution
     There is little available data on the absorption or excretion
of 1,2-diphenylhydrazine by mammals.  The administration of DPH by
various routes results in systemic effects and the presence of DPH
metabolites in the urine, indicating some degree of absorption.
Metabolism
     The metabolism of 1,2-diphenylhydrazine in the rat  is present-
ed in Figure 1 (Williams, 1959).  The compound was administered to
rats orally (p.o.) (200, 400 mg/kg), intraperitoneally  (i.p.)  (200
mg/kg), intratracheally (5,  10  mg/kg}, and intravenously (i.v.)  (4,
8 mg/kg).   Urine was  analyzed chromatographically  (TLC)  and the
metabolic  scheme  shown in Figure  1  was proposed.   Benzidine was
identified  as  a  metabolite.   The  metabolites detected  were not
dependent upon the dose or the route of administration.
                             EFFECTS
Acute, Subacute, and Chronic Toxicity
     Two studies  reporting  oral  LD5Q values  for  DPH were identi-
fied.  Marhold, et al.  (1968)  used 10  male  Wistar rats and admin-
istered DPH as a 5 percent aqueous suspension.  The LD^Q value was
reported to be 959 mg/kg.  Liver damage is an important feature of
diphenylhydrazine toxicity,  particularly after  chronic exposure.
Sutton  (1967)  noted  that phenylhydrazines  cause  prominent kidney
damage and more diffuse liver damage in animals.
     No  epidemiological  studies  of  DPH  exposure have  been con-
ducted.
                               C-3

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                                                    OH
          NHAC
                   FIGURE 1
1,2-Diphenylhydrazine Metabolites in Rat Urine
            Source: Williams, 1959
                       C-4

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Synergism and/or Antagonism
     Marhold, et  al.  (1968)  demonstrated that diphenyline, a pro-
duct of benzidine  rearrangement,  acts synergistically with benzi-
dine to produce tumors.  Genin  (1975) showed synergism of  hydrazo-
benzene with  benzidine  sulfate.   Kurlyandskii,  et  al.  (1976) ob-
served that benzidine sulfate-hydrazobenzene or  benzidine  sulfate-
dianisidine  sulfate  mixtures when  administered  subcutaneously to
rats increased  the incidence of  bladder  cancer  and decreased the
latent period for  tumor  development  when compared with the  carcino-
genic activity of the individual compounds.   The  authors emphasized
the  importance  of preventing the possible  exposure of industrial
workers to combinations of 1,2-diphenylhydrazine  and benzidine dur-
ing the manufacture of benzidine sulfate.
Teratogenicity
     Pertinent data  could  not be located in the available litera-
ture on the teratogenicity of DPH.
Mutagenicity
     Sieler  (1977) studied  the  incorporation of  H-thymidine into
testicular DNA using the technique developed by  Friedman and Staub
(1976).  When DPH was administered i.p. in a dose of 100 mg/kg, an
inhibitory effect  on testicular  DNA synthesis  was observed.   In
this study, a limited number of  structurally related compounds were
assayed.  The relationship  between  the  known mutagenicity of some
of these agents agreed favorably with effects on  depressed  testicu-
lar DNA synthesis.  These authors then concluded that agents which
can depress  testicular  DNA synthesis may  have a mutagenic poten-
tial.
                               C-5

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Carcinogenicity
     Concern  over  1,2-diphenylhydrazine is  based  on several  fac-
tors:   (1) its presence in drinking water  (at concentrations up to
1 yg/1  (U.S.  EPA,  1975);  (2)  the likely  production of benzidine
from  DPH  in  the stomach  due   to  gastric  acidity  [International
Agency  for  Research  on  Cancer  (IAJRC) ,  1972];  (3) the documented
carcinogenicity  of  diphenylhydrazine  and  selected   substituted
hydrazines  (IARC,  1974);  (4)  the  increased incidence  of bladder
cancer among  workers  involved  in the  manufacture  of dyes  (Wynder,
et  al.  1963;  Anthony,  et al.  1970);  (5)  the  carcinogenicity of
azobenzene, a  metabolite  of  DPH, as well  as the  established  car-
cinogenicity of benzidine to which DPH  is  converted.
     There are several reports  on  the possible carcinogenicity of
DPH.   In a  study by  the  National Cancer  Institute (NCI, 1978),
technical grade DPH* was fed as  a dietary  admixture  to  Fischer 344
rats  and  mice (B6C3F1)  of both  sexes.   Dietary concentrations of
DPH fed to rats and mice are indicated  in  Tables 1 and  2.  In  this
study, DPH was fed to mice and  rats for 78  weeks followed by obser-
vation periods of 17 to 96 weeks and 28 to  109 weeks, respectively.
Controls consisted of 47 to 50  animals of each  sex.   The results of
this  study are summarized in Tables 1  and 2.   There were differ-
ences in the nature and organ distribution of tumors between sexes
and species.   DPH  was  carcinogenic to  Fischer 344 rats  of  both
sexes and caused significant increases  in  hepatocellular carcinoma
*mp » 120 - 124°C  (K & K labs)
                               C-6

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                                           TABLE 1

                  Carcinogenicity of  1,2-Diphenylhydrazine  in B6C3F-^  Mice*
Sex

Male


Female

1
50
50
50
50
50
50
47
50
Dosea
LD control
HD control
0.008
0.040
LD control
HD control
0.004
0.040

Treated
0
0
78
78
0
0
78
78
Weeks
Observed
Post Treatment
95
96
17
17
96
96
17
18
Effects
Hepatocellular
Carcinomas
12/50
6/48
11/47
8/46
2/47
1/50
4/39
20/43 p^O.OOl
Pulmonary
Carcinomas
5/50
5/49
1/47
0/46
2/46
3/50
3/38
2/40
*Source: NCI, 1978.
aDose = % DPH in diet.
 LD = Low dose.
 HD = High dose.
                                       C-7

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 Sex
Male
                                                        TAHLE 2

                             Carcinogenic!ty of 1,2-Diphenylhydrazine in Fischer 344 Rats*
Weeks
1
50
49
50
50
50
50
50
50
Dose3
LO control
HD control
0.008
0.03
LD control
HD control
0.004
0.010
Treated
0
0
78
78
0
0
78
78
Observed
108
109
29
28
109
109
30
29
llepatocellular
Carcinomas or
Neoplastic
Nodules
5/47
1/48
13/49 p = 0.040
37/49 p 
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or  neoplastic  nodules in male  rats  at both doses; Zymbal's  gland
squamous-cells or adrenal tumors in male rats at the high dose;  and
neoplastic  liver  nodules or mammary  carcinomas in female  rats at
the  high  dose.   Female  mice showed  an  increase in  hepatocelluar
carcinomas  only at  the  high dose.   DPH was  not carcinogenic  in
B6C3F1 male mice.
     In a study by Pliss (1974) , the carcinogenic properties of  DPH
were studied over  a  period  of 588 days  in  rats (N = 163)  and  C57
mice (N = 110).  DPH was suspended in sunflower seed oil  and admin-
istered  by  subcutaneous  (s.c.)   injection  (40  mg/wk/rat  and  5
mg/wk/mouse), and by  addition to  food  (30 mg/5  times/wk)  or appli-
cation to the skin (2 mg/3 times/wk/mouse).
     The  data  summarized in Table  3  indicate  that DPH  produces  a
wide variety of tumors in both mice and  rats.
     In contrast  to  the  studies  of NCI  (1978)  and Pliss  (1974),
Marhold, et al. (1968) and Spitz (1950) did  not find DPH  to  be car-
cinogenic.  The latter two studies were  difficult  to  interpret  due
to the lack of  specific  information on the  purity of DPH,  experi-
mental design,  or  statistical  analysis.  The  Pliss study  (1974)
should be used in a cautious manner in  indicting DPH  as  a carcino-
gen.  The author indicated that  animals had to be added to the study
in order  to replace  animals  afflicted with  a parasitic  infection.
In addition, although the tumor  incidence was given for DPH-treated
animals,  the incidence  of tumors in control  animals  was not pre-
sented except  in  the case  of the  epicutaneous administration  of
DPH.  Values  of 17 percent  vs.  22.2  percent  for  control  and  DPH
                                 C-9

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                              TABLE 3

              Carcinogenicity  of  DPH  in  Mice  and  Rats*
Species
                                             Effects
    Route
 % Tumor
Incidence
Tumors
 Mice
 Rats
    s.c.

    P.O.


epicutaneous

    s.c.
   36.6     Rhabdomyosarcoma

   50       Pulmonary adenoma,
             leukemia, liver

   22.2     Skin, lung, liver

   22.6     Uterus, mammary,
             Zymbal's gland, liver,
             spleen, lymphoid leukemia
*Source: Pliss, 1974.
                               C-10

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groups, respectively, were presented but no statistical analysis of
these incidences was given.   In  light of  the available information,
the NCI data indicates that DPH is carcinogenic.
                                 C-ll

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                      CRITERION FORMULATION
Existing Guidelines and Standards
     Existing guidelines  or  standards were not  found for 1,2-di-
phenylhydrazine.
Current Levels of Exposure
     Pertinent data could not be located in the available litera-
ture on  the  concentration of  1,2-diphenylhydrazine  in the atmos-
phere.
     1,2-Diphenylhydrazine has been found to be present in drinking
water at levels of 1 ug/1 » 1 ppb  (U.S. EPA, 1975).
     1,2-Diphenylhydrazine has not been found  to be a  natural con-
stituent of food.
Special Groups at Risk
     Manufacturers of dyes and Pharmaceuticals are subject to occu-
pational exposure.  Groups working in the laboratory  and forensic
medicine may also be subject  to  1,2-diphenylhydrazine  exposure.
Basis and Derivation of Criterion
     An  evaluation  of  the  subacute, acute  and chronic  toxicity,
with the exception  of carcinogenicity  is impossible because of
scanty  data.   Guidelines or  standards  presently do not exist  for
DPH.  Diphenylhydrazine has been shown to  produce  carcinogenic  re-
sponses  in rats  and  mice  (NCI, 1978; Pliss, 1974).  Since the  NCI
(1978)   study represents  the only report in which all  the data  can
be  analyzed,  it will  be  used as  a basis  for formulating  a  cri-
terion.
     Under  the  Consent Decree  in  NRDC v.  Train,  criteria are to
state  "recommended  maximum permissible  concentrations  (including
                               C-12

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where appropriate, zero) consistent with the protection of aquatic
organisms, human health, and recreational activities."  DPH is sus-
pected of being a human carcinogen.  Because  there  is no recognized
safe concentration for  human carcinogens,  the recommended concen-
tration of DPH  in water for maximum protection of human health is
zero.
     Because attaining a zero concentration level may be infeasible
in some cases and  in  order  to assist  the  Agency and states in the
possible future development of water quality regulations, the con-
centrations of  DPH corresponding to  several incremental lifetime
cancer risk levels have been estimated.   A cancer risk level pro-
vides an estimate of  the additional  incidence of  cancer that may be
expected  in  an exposed  population.   A risk  of  10   for example,
indicates a probability of one additional case of cancer for every
100,000 people  exposed, a  risk  of  10~6  indicates one additional
case of cancer for every million people exposed, and so forth.
     In the Federal Register notice of availability of draft ambi-
ent water quality criteria, EPA stated that  it is considering set-
ting criteria  at an  interim target risk  level  of  10   ,  10" , or
10   as shown in the  following information.
Exposure Assumptions
2 liters of drinking
water and consumption
of 6.5 g fish and
shellfish (2)
Consumption of fish
and shellfish only
             Risk Levels
   and Corresponding Criteria (1)
Q
0
 io-7
4 ng/1
 IP"6
42 ng/1
 IP"5
422 ng/1
    56 ng/1    560 ng/1  5,600 ng/1
                               C-13

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(1)  Calculated by  applying  a  linearized multistage model as dis-
     cussed in the Human Health Methodology Appendices to the Octo-
     ber 1980  Federal  Register  notice which  announced  the avail-
     ability of this document.  Appropriate  bioassay data used in
     the calculation of  the model are presented  in the Appendix.
     Since  the extrapolation model  is linear  to low  doses,  the
     additional lifetime risk is directly proportional to the water
     concentration.   Therefore, water concentrations corresponding
     to other risk levels  can be derived  by multiplying  or dividing
     one of the risk levels and corresponding water concentrations
     shown  in  the table  by factors such  as  10,  100,  1,000 and so
     forth.
(2)  Seven percent of the  DPH exposure results from the  consumption
     of aquatic organisms  which exhibit an average bioconcentration
     potential of 24.9-fold.   The  remaining 93 percent of DPH expo-
     sure results from drinking water.
     Concentration levels were derived assuming a  lifetime exposure
to various  amounts  of  DPH,  (1)  occurring from the  consumption of
both drinking water and aquatic life grown in water containing the
corresponding  DPH concentrations  and,   (2)  occurring solely from
consumption of aquatic life grown  in  the  waters containing  the cor-
responding DPH concentrations.
     Although  a total  exposure  evaluation  for DPH  is desirable
there is no data  to support a total  exposure analysis.  The cri-
teria presented, therefore, assume an incremental  risk from assumed
ambient water exposure only.
                               C-14

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     For DPH the  case  for  criterion development is based upon the
existence of carcinogenicity responses in animals  (rats and mice).
     Because of  the  lack of investigations  for other chronic and
acute responses,  no  information on  other  effects exists in either
human or  animal  systems.  Thus,  the  criterion proposed should be
considered as precautionary  until further studies can  be  used in
the overall toxicity evaluations.
                                C-15

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                            REFERENCES

Anthony, A.M.,  et al.  1970.   Tumors  of  the  urinary bladder:  An
analysis of  the  occupations of 1,030 patients  in  Leeds, England.
Jour. Natl. Can. Inst.  45: 879.

Friedman, M. and  J.  Staub.   1976.   Inhibition of mouse  testicular
DNA synthesis by mutagens and carcinogens as a potential mammalian
assay for mutagenesis.  Mutat. Res.  37: 67.

Genin, V.A.  1975.  Increase in carcinogenic activity during joint
effect of  hydrazobenzene  and  benzidine  sulfate.   Gig.  Tr.  Prof.
Zabol.  6:  28.

Hansch, C.  and  A.J. Leo.   1979.  Substituent Constants for Correla-
tion Analysis  in Chemistry and Biology.   Wiley-Interscience,  New
York.

International  Agency for  Research  on  Cancer.   1972.   Aromatic
amines.  Monograph on the  evaluation  of carcinogenic risk of chemi-
cals to man.  1: 69.

International Agency  for Research on Cancer.  1974.  Hydrazine and
its derivatives.  Monograph on the evaluation of carcinogenic risk
of chemicals to man.  4: 81.
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Kurlyandskii, B.A.,  et  al.   1976.   Experimental study on the com-
bined effect of  some  diphenylamino  derivatives  with regard to the
prevention of occupational urinary bladder growths.   Gig. Tr. Prof.
Zabol.  5: 34.

Marhold,  J.,  Jr.,  et al.   1968.   The possible  complicity of di-
phenyline in the origin of tumors in the manufacture of benzidine.
Neoplasma.  15: 3.

National  Cancer  Institute.   1978.   Bioassay of hydrazobenzene for
possible carcinogenicity.  Publication NO.  (NIH) 78-1342.

Pliss,  G.B.    1974.   Carcinogenic  properties  of  hydrazobenzene.
Vop. Onkol.  20: 53.

Sieler, J.P.    1977.   Inhibition of  testicular DNA synthesis by
chemical  mutagens  and  carcinogens.    Preliminary  results  in  the
validation of a novel short term test.  Mutat. Res.  46: 305.

Spitz, S.   1950.   The carcinogenic action  of  benzidine.   Cancer.
3: 789.

Stephan C.E.  1980.  Memorandum to J.  Stara.  U.S.  EPA.  July 3.

U.S. EPA.  1975.   Preliminary  assessment  of suspected carcinogens
in drinking water.  Rep. to Congress.  11.
                               C-17

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U.S.  EPA.    1980.   Seafood  consumption data  analysis.   Stanford



Research Institute  International,  Menlo Park,  California.   Final



rep., Task II.  Contract 68-01-3887.







Veith, G.D.  1980.  Memorandum to C.E. Stephan.  April 14.







Veith, G.D., et al.   1979.  Measuring  and estimating  the bioconcen-



tration factor of chemicals  in  fish.   Jour.  Fish.  Res. Board Can.



36: 1040.







Williams, R.  1959.   Detoxication Mechanisms.   John Wiley  and Sons,



New York.  p. 480.







Wynder, E.L.,  et  al.  1963.  An epidemiological investigation of



cancer in the bladder.  Cancer.  16:  1388.
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                             APPENDIX
              Summary and Conclusions Regarding the
            Carcinogenicity  of  1,2,-Diphenylhydrazine*
     1,2-Diphenylhydrazine is  used  primarily in dye manufacturing
industries as a precursor in  the  synthesis of benzidine.  There are
no  data  showing carcinogenic  effects  of 1,2-diphenylhydrazine in
humans.  However,  two studies have shown  that 1,2-diphenylhydrazine
is  carcinogenic in mice  and  rats via subcutaneous and oral routes
of administration.  Male rats, receiving dietary concentrations of
0.03 percent  1,2-diphenylhydrazine,  developed  hepatocellular car-
cinomas, squamous cell carcinomas of the Zymbal's gland, and adre-
nal tumors.  Female rats, receiving  0.01  percent 1,2-diphenylhydra-
zine in  the  diet,  developed neoplastic  nodules of  the  liver  and
mammary adenocarcinomas.   Female  mice,  exposed  to  0.04 percent 1,2-
diphenylhydrazine  in  the  diet,  developed  hepatocellular  car-
cinomas.
     The carcinogenic responses induced in male and  female rats and
female mice  constitute substantial  evidence that 1,2-diphenylhy-
drazine is likely to be a human carcinogen.
     The water quality criterion  for  1,2-diphenylhydrazine is based
on the induction of hepatocellular carcinoma and neoplastic nodules
in male Fischer 344  rats, exposed to a time-weighted average con-
centration of  0.008  or 0.03  percent (80  or  300 ppm) 1,2-diphenyl-
hydrazine in the diet for 78 weeks  (NCI, 1978).  The concentration
of  1,2-diphenylhydrazine  in  water calculated to keep the lifetime
cancer risk below 10"5 is 0.42 ug/1.
*This  summary  has  been  prepared  and approved  by the Carcinogens
 Assessment Group of EPA on June 15, 1979.
                               C-19

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                    Summary of  Pertinent Data







     The water quality criterion for 1,2-diphenylhydrazine is based



on the induction  of  hepatocellular carcinomas and  neoplastic nod-



ules in male Fischer 344 rats, exposed to various concentrations of



1,2-diphenylhydrazine  in the diet ad libitum  for  78  weeks (NCI,



1978) .   The criterion was calculated from the  following  parameters:





        Dose                          Incidence

     (mg/kg/day)               (no.  responding/no,  tested)



           0                              6/95



           4                             13/49



          15                             37/49







     le =  78 weeks           w =  0.380  kg



     Le = 104 weeks           R =  24.9 I/kg



     L  = 104 weeks





     With  these  parameters  the carcinogenic potency  for  humans,



q *, is 0.768  (mg/kg/day)   .  The  resulting water concentration of



1,2-diphenylhydrazine,  calculated  to  keep  the  individual  lifetime


                    — 5
cancer risk below 10   ,  is  0.42 yg/1.
                               C-20
                                           U. S. GOVERNMENT PRINTING OFFICE 1MO 720-C16/4377

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