DIPHENYLHYDRAZINE
Ambient Water Quality Criteria
              Criteria and Standards Division
              Office of Water Planning and Standards
              U.S. Environmental Protection Agency
              Washington, D.C.

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                        CRITERION DOCUMENT


                      1,2-DIPHENYLHYDRAZINE


CRITERIA


                           Aquatic Life


     For 1,2-diphenylhydrazine the criterion  to protect  freshwater


aquatic life as derived using the Guidelines  is 17  ug/1  as  a 24-


hour average and the concentration should not  exceed  38  ug/1 at


any time.


     For saltwater aquatic life, no criterion  for 1,2-diphenyl-


hydrazine can be derived using the Guidelines, and  there are


insufficient data to estimate a criterion using other procedures.


                           Human Health


     For the maximum protection of human health from  the potential


carcinogenic effects of exposure to 1,2-diphenylhydrazine through


ingestion of water and contaminated aquatic organisms,  the  ambient


water concentration is zero.  Concentrations of 1,2-diphenylhydra-


zine estimated to result in additional lifetime cancer risks rang-


ing from no additional risk to an additional  risk of  1 in 100,000

                                                              i
are presented in the Criterion Formulation section  of this  docu-


ment.  The Agency is considering setting criteria at  an  interim


target risk level in the range of 10~5, 10~~6,  or 10"? with  cor-


responding criteria of 0.4 ug/1/ 0.04 ug/1 and 0.004  ug/1/  re-


spectively.

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Introduction
_^MHMH^WB^_^V^^BB                 ^
     Diphenylhydrazine exists as an assymmetrical isomer, 1,1-
diphenylhydrazine, and a symmetrical isomer, 1,2-diphenylhy-
drazine (hydrazobenzene).  The hydrochloride of 1,1-diphenyl-
hydrazine is used as a reagent for the sugars, arabinose and
lactose (Windholtz, 1976).  1,2-Diphenylhydrazine is used in
organic synthesis (Bennett, 1974) and has a major use as the
starting material in the production of benzidine, for use in
dyes (Lurie, 1964).
     No data were found on the environmental presence or per-
sistence of diphenylhydrazines, except for one report of de-
tection in drinking water at a concentration of 1 ug/1  (U.S.
EPA, 1975).  1,1- and 1,2-diphenylhydrazines have been  char-
acterized as slightly soluble and insoluble in water, respec-
tively (Windholtz, 1976; Bennett, 1974).  No quantitative data
were found for the water solubilities and vapor pressures of
these compounds; consequently, no predictions can be made
about their presistence in water,.
     Since no data are available on levels of diphenylhydra-
zines in surface waters, ambient water levels cannot be com-
pared to the levels of these chemicals determined to be toxic
in laboratory studies.  Experimental data indicate that 1,2-
diphenylhydrazine is toxic to freshwater aquatic organisms and
reported LC50 values range from 0.27 to 4.1 mg/1 (U.S.  EPA,
1978).   Rats and mice developed a variety of malignancies
after dermal exposures to 1,2-diphenylhydrazine (Spitz, et al.
1950; Pliss, 1974).  Oral doses of 1,2-diphenylhydrazine did
                             A-l

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not produce malignancies  (Marhold, et al. 1968).  The docu-
mented carcinogenicity of 1,2-diphenylhydrazine  is of primary
concern to industrial workers, who are exposed to higher con-
centrations than other segments of the population.
     Although there are few data describing the  fate of di-
phenylhydrazines in water, the carcinogenic effects of 1,2-di-
phenylhydrazine warrant its regulation, to protect humans and
aquatic organisms from possible water-related hazards associ-
ated with this chemical.
                             A-2

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                          REFERENCES

Bennett, H., ed. 1974.  Concise chemical and technical dic-
tionary.  Chemical Publishing Co. Inc., New York.

Keinath, T.M. 1976.  Benzidine:  Wastewater treatment tech-
nology.  Prepared for Off. Water Plann. Stand. U.S. Environ.
Prot. Agency.

Lurie, A.P. 1964.  Benzidine. P. 408. Ijn  Kirk-Othmer Ency-
clopedia of Chemical Technology. 2nd ed. Vol. 3.

Marhold, J., et al. 1968.  The possible complexity of di-
phenyline in the origin of tumors in the manufacture of ben-
zidine.  Neoplasma 15: 3.

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

Spitz, S., et al. 1950.  The carcinogenic action of benzi-
dine.  Cancer Sept: 789.

U.S. EPA. 1975.  Preliminary assessment of suspected carcino-
gens in drinking water.  Off. Tox. Subs. Washington, D.C.

U.S. EPA. 1978.  In-depth studies on health and environmental
impacts of selected water pollutants.  Contract No.  68-01-  .
4646.
                             A-3                                  7

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Windholtz, M., ed. 1976.  The Merck Index. Merck and Co,
Inc., Rahway, N.J.
                             A-4

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AQUATIC LIFE TOXICOLOGY*

                       FRESHWATER ORGANISMS

Introduction

     Toxicity tests with the bluegill  and Daphnia  magna have been

conducted using static procedures, and the results demonstrated
   4
adverse effects as low as 251 ug/1.

Acute Toxicity

     The unadjusted 96-hour LC50 for the bluegill  is  270 ug/1

(Table 1), and when this concentration is adjusted for  testing
   ' V
procedures and species sensitivity a Final Fish  Acute Value  of 38

ug/1 is obtained.

     Daphnia magna is less sensitive with an  unadjusted 48-hour

EC50 of 4,100 ug/1 (Table 2).  The resultant  Final Invertebrate
  j
Acute Value of 170 ug/1 is higher than that value  for fish,  so the

latter, 38 ug/1, becomes the Final Acute Value.
*The reader is referred to the Guidelines for Deriving Water
     ;
Quality Criteria for the Protection of Aquatic Life  [43  FR  21506

(May 18, 1978) and 43 FR 29028 (July 5, 1978)] in  order  to  better

understand 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  the  calcula-

tions for deriving various measures of toxicity as described  in

the Guidelines.
                             B-l

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Chronic Toxicity

     The chronic effects of 1,2-diphenylhydrazine on Daphnia magna

have been determined  (U.S. EPA, 1978) and adverse effects were

observed at 420 ug/1  (Table 3).  No detectable effects occurred ,at

150 ug/1.  The latter  concentration is 0.037 of the 48-hour EC50

(Table 2).  A Final Invertebrate Chronic Value of 49 ug/1 is

obtained from these results and, since there are no data on

chronic toxicity to fish, plant effects, or Residue Limited

Toxicant Concentration, this concentration also becomes the Final

Chronic Value.

Residues

     No measured steady-state bioconcentration factor  (BCF) is

available for 1,2-diphenylhydrazine.  A BCF can be estimated using

the octanol-water coefficient of 870.  This coefficient is used to

derive an estimated BCF of 100 for aquatic organisms that contain

about 8 percent lipids.  If it is known that the diet of the

wildlife of concern contains a significantly different lipid

content, an appropriate adjustment in the estimated BCF should be
                                                                X
made.
                             B-2

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CRITERION FORMULATION
                     Freshwater-Aquatic Life
Summary of Available Data
     The concentrations below have been rounded  to  two  significant
figures.
     Final Fish Acute Value = 38 ug/1
     Final Invertebrate Acute Value = 170 ug/1
          Final Acute Value = 38 ug/1
     Final Fish Chronic Value = not available
     Final Invertebrate Chronic Value = 49 ug/1
     Final Plant Value - not available
     Residue Limited Toxicant Concentration  not available
          Final Chronic Value = 49 ug/1
          0.44 x Final Acute Value = 17 ug/1
     The maximum concentration of 1,2-diphenylhydrazine  is the
Final Acute Value of 38 ug/1 and the 24-hour average  concentration
is 0.44 times the Final Acute Value.  No important  adverse effects
on freshwater aquatic organisms have been reported  to be caused  by
concentrations lower than the 24-hour average concentration.
     CRITERION:  For 1,2-diphenylhydrazine the criterion to
protect freshwater aquatic life as derived using the  Guidelines  is
17 ug/1 as a 24-hour average and the concentration  should not
exceed 38 ug/1 at any time.
                             B-3

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                         Table  1    Freshwater fish acute values for 1,2-diphenylhydrazine  (U.S.  EPA,  1978)


                                                                            Adjusted
                                   Bioassay  Test      Time      LC50      LC50
           Organise                Method*   Cone.**    (fare)-     (uq/11     (ug/ll


           Bluegill.                  S        U        96        270       150
           Lepomis macrochirus
           *  S = static

           ** U  unmeasured

              Geometric mean of adjusted  values = 150  ug/1        =  38  pg/1
00
I

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09
I
tn
                         Table  2.   Freshwater invertebrate acute values for 1,2-diphenylhydrazine (U.S. EPA, 1978)
Organism
Cladoceran,
Daphnia magna
fiioassay
MetiiOQ*
S
Test
Cone .**
U
Time
(nts)
48
Adjusted
LC50 LCbO
(Uq/i) (Uq'l)
4,100 3,470
           *  S = static


           ** U = unmeasured


              Geometric mean of adjusted  values = 3,470 pg/1    A.   - 170 wg/1

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                     Table 3.  Freshwater invertebrate chronic values for 1,2-diphenylhydrazine (U.S. EPA, 1978)

                                                                 \
                                                             Chronic
                                                   Limits    Value
            Organism                     Teat*     lug/11     (yg/1)


            Cladoceran,                    LC       150-420     251
            Daphnla  magna
            *  LC    life  cycle  or  partial  life  cycle

              Geometric  mean of chronic value

              Lowest  chronic value =251  Mg/1
                                              251
Geometric mean of chronic values - 251 ug/1    -  * 49 wg/1
Cfl

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                       SALTWATER ORGANISMS

Introduction


     No acute toxicity, chronic toxicity or residue data are
     t

available for saltwater organisms and 1,2-diphenylhydrazine.
                             B-7

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CRITERION FORMULATION
                       Saltwater-Aquatic  Life
     No saltwater criterion can be derived  for  1,2-diphenylhy-
drazine using the Guidelines because  no  Final Chronic Value  for
either fish or  invertebrate species or a good substitute  for
either value is available, and there  are insufficient data to
estimate a criterion using other procedures.
                             B-8

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                    1,2-DIPHENYLHYDRAZINE



                         REFERENCES







U.S. EPA. 1978.  In-depth studies on health and environmental



impacts of selected water pollutants. Contract No. 68-01-



4646.
                              B-9

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-Mammalian Toxicology  and Human  Health  Effects
                           EXPOSURE
 Introduction
      1,2-diphenylhydrazine  (DPH)  is  a  precursor  in the manu-
 facture of  benzidine, an intermediate  in the production
 of dyes.  If2-diphenylhydrazine is used in the synthesis
 of phenylbutazone,  a  potent  anti-inflammatory (anti-arthritic)
 drug  (Wenner,  1967).
      The commercial production  of 1,2-diphenylhydrazine
 per se in 1977 was  in excess of 1000 Ibs.  (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  and a contaminant but there is no way
 of estimating  these quantities  which are substantial.
      Industrial  exposure to  1,2-diphenylhydrazine is prim-
 arily limited  to workers in  the dye  manufacturing industry
 and in the  pharmaceutical  industry.
      Derivatives of hydrazine are reported to be hepatotoxic,
 hemolytic,  convulsants, and  irritants.   They are absorbed
 from  all routes  (Button, 1967).
      The basis for  concern over 1,2-diphenylhydrazine includ-
 es:   (1) its presence in drinking water (highest concentra-
 tion  reported  was  1 pg/1  (Ippb),  U.S.  EPA, (1975); (2) the likely
 production  of  benzidine from DPH in  the stomach  due to gas-
 tric  acidity  (IARC, 1972);  (3)  the documented carcinogenicity
 of hydrazine and selected substituted  hydrazines (IARC,
 1974); (4)  the increased incidence of  bladder cancer among
 workers involved in the manufacture  of  dyes (Wynder, et al.
                             C-l

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1963); Anthony, et al. 1970);  (5) the carcinogenicity of
azobenzene, a metabolite of DPH, as well as the established
carcinogenicity of benzidine to which DPH is converted.
     The structure and physical data for 1,2-diphenylhydra-
zine are presented in Figure 1.
     The primary commercial method of production of this
compound is the reduction of nitrobenzene by catalysts such
     j-3      +2
as Fe   or Zn   in alkaline solution.  By this procedure,
one obtains nearly quantitative yields  (Kirk-Othmer, 1963).
Figure 2 depicts this process  as well as the by-products
of diphenylhydrazine, azobenzene and azoxybenzene.
     The reaction of 1,2-diphenylhydrazine with acid results
in the benzidine rearrangement  (Kenner, 1968).  This reaction
is presented in Figure 3.  In  addition to benzidine, other
products formed include diphenyline, o-benzidine, and o-
semidine.  In the stomach, 1,2-diphenylhydrazine can be
converted into benzidine, a human carcinogen  (Haley, 1975;
IARC, 1972).
                             C-2

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               Synonyms -  Hydrazobenzene
                           Symmetrical diphenylhydrazine
                           N,N'  Diphenylhydrazine
                           N  N'  - Bianiline
                           1,1'  Hydrazodibenzene
               Cas No.      530-50-7(b)
                           Molecular weight = 184.24
                           Melting Point =    131C
                           Boiling Point =    220C
               Solubility =     Slightly soluble in water
                                Very soluble in benzene, ether, alcohol
Figure 1.  1,2-diphenylhydrazine:  Chemical and Physical Properties3
(a)  from CRC Handbook - 59th Ed.
(b)  DHEW report,  1978
                              C-3

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                                         Oi
Nitrobenzene
                              Alkaline
                                  "1"^
          *Azobenzene
*Azoxybenzene
                            H H
                                              1,2-diphenylhydrazine
Figure 2. Synthesis of 1,2-diphenylhydrazine  (Williams, 1959)
*Carcinogenic
                               C-4

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                      H H
1,2-diphenylhydrazine
                                             Diphenyline
                               o-Benzidine
               o-Seraidine
Figure 3. Benzidine Rearrangement of 1,2-diphenylhydrazine
          (Williams, 1959)
*Carcinogenic
                            c-5

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Ingestion from Water and Foods
     DPH was not detected in the finished water of any of
ten cities selected for a detailed EPA study in 1975. However,
the same study demonstrated the presence of DPH in drinking
water in concentrations up to 1 jug/1 (1 ppb).
  '   There are no available data identifying DPH as a direct
or indirect food additive, or as a naturally occurring consti-
tuent of any food.
Inhalation, Dermal, and Other Sources
     Workers involved in the manufacture of dyes, certain
Pharmaceuticals and chemicals, laboratory workers, and work-
ers in forensic medicine risk occupational exposure to 1,2-
diphenylhydrazine.  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.
                     ;  PHARMACOKINETICS
Absorption, Excretion, and Distribution
     There are no available data on the absorption or excre-
tion 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.  This indicates
that it is absorbed.
Metabolism
     The metabolism of 1,2-diphenylhydrazine in the rat
is presented in Figure 4 (Williams, 1959).  The 1,2-diphenyl-
hydrazine was administered to rats orally  (200, 400 mg/kg),
i.p. (200 mg/kg), intratracheally  (5, 10 mg/kg), and i.v.
(4, 8 mg/kg).  Urines were analyzed chromatographically
                              C-6

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Route
p.o.
i.p.
i.t.a
i. v.
Dose (mg/kg)
 200, 400
 100, 200
  5, 10
  4, 8
Solvent
  oil
 DMSOb
 water
 DMSO
                                                            NH,
Figure 4. 1,2-diphenylhydrazine Metabolites  in Rat Urine
          (Williams, 1959)
 intratracheally
Dimethyl sulfoxide
                               C-7

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(TLC) and a scheme proposed  (Figure 4). Benzidine was identi-
fied as a metabolite.  The metabolites detected were not
dependent upon the dose or the route of administration.
                           EFFECTS
Acute/ Sub-acute, and Chronic Toxicity
     Two studies reporting DPH oral LD^Q'S were identified.
Marhold, et al. (1968) used  ten male Wistar rats and adminis-
tered DPH as a five percent  aqueous suspension.  The LD
reported was 959 mg/kg. In the Registry of Toxic Effects
of Chemical Substances, 1977  (RTECS) the oral LD5Q is listed
as 301 mg/kg. No details were given.  The basis for this
difference is not known. Liver damage is an important feature
of hydrazine toxicity, particularly after chronic exposure,
but Sutton (1967) noted that phenylhydrazines cause prominent
kidney damage and more diffuse liver damage in animals.
     Two rat studies were reported in Reg. Tox. Eff. Chem.
Subst.  (1977) .  There were no data available on the strain,
sex, doses, or solvents used for these studies.  One rat
study reported an oral TDLo  of 396 gm/kg. Neoplasms were
seen by 53 weeks. In the second rat study, a total dose
of 16 mg/kg DPH was administered subcutaneously and neoplasms
were reported after 52 weeks.
     Two mouse studies were  reported in Reg. Tox. Eff. Chem.
Subst.  (1977).  When 1,2-diphenylhydrazine was applied topi-
cally for 25 weeks (total dose of 5,280 mg/kg) neoplasms
were found.  Subcutaneous injection of DPH  (8,400 mg/kg)
resulted in neoplasms after  38 weeks.  These results are
presented in Table 1.
                               C-8

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                                                             TABLE 1
                                               1,2-diphenylhydrazine Toxicity Data
o
Species/Strain
Rat/Wistar
Rat/N.O.S.d
Rat/N.O.S.
Rat/N.O.S.
Mouse/N.O.S.
Mouse/N.O.S.
Sex No. Dose Solvent Cone Route T.I.a Duration
(mg/kg)
M 10 N.O.S. water 5% p.o. U>SQ 959 N.O.S.
(Suspension)
N.O.S. N.O.S. N.O.S. N.O.S. N.O.S. p.o. U>50 301 N.O.S.
N.O.S. N.O.S. N.O.S. N.O.S. N.O.S. p.o. TDLoe 53 wks.
39,600
N.O.S. N.O.S. N.O.S. N.O.S. N.O.S. s.c.C TDLo 52 wks.
16,000
N.O.S. N.O.S. N.O.S. N.O.S. N.O.S. skin TDLo 25 wks.
5,280
N.O.S. N.O.S. N.O.S. N.O.S. N.O.S. S.C. TDLo 38 wks.
Effects
N.O.S.
N.O.S.
Neoplasms
Neoplasms
Neoplasms
Neoplasms
References
Mar hold, et al.
(1968)
RTECSb
RTECS
RTECS
RTECS
RTECS
                                                                            8,400
    (a)  Toxicity  Index
    (b)  Registry  of Toxic Effects of Chemical Substances, 1977
    (c)  Subcutaneous
    (d)  Not  Otherwise Specified
    (e)  TDLo-Toxic Dose Low - the lowest dose of a substance introduced by any route, other than
        inhalation, over any given period of time and reported to produce any toxic effect in
        humans or to produce carcinogenic, teratogenic, mutagenic, or neoplastigenic effects in
        humans of animals.

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     No epidemiological studies have been carried out on
the effects of DPH in humans.
Synergism and/or Antagonism
     Marhold, et al.  (1968) demonstrated that diphenyline,
a product of benzidine rearrangement, acts synergistically
with benzidine to produce tumors. Genin  (1975) showed a
synergistic effect of hydrazobenzene and benzidine sulfate.
Kulyanskii, et al. (1976) observed 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
emphasize the importance of preventing the possible exposure
of industrial workers to combinations of 1,2-diphenylhydra-
zine and benzidine during the manufacture of benzidine-sulfate,
Teratogenicity
     There are no studies available 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).  OPH was administered  i.p. in a dose of
100 mg/kg and an inhibitory effect on testicular DNA synthesis
was observed. Seiler  implied from this study that DPH has
a mutagenic potential.
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Carcinogenicity
     There are several reports on the possible carcinogeni-
city of DPH.  The most comprehensive study was reported
in 1978 (NCI, 1978).  Technical grade DPH* was fed as a
dietary admixture to rats  (Fischer 344) and mice  (B6C3F1)
of both sexes.  Dietary concentrations  (DPH) fed  to rats
and mice are indicated in Tables 2 and  3.  DPH was fed to
mice and rats for 78 weeks followed by  observation 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  2 and 3.   There were
differences 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 at 5 pg/kg/day and 18.8 jig/kg/day;
Zymbal's gland squamous-cell tumors in  male rats  at 18.8
jug/kg/day and neoplastic liver nodules  in female  rats at
7.5 jug/kg/day.  Female mice showed an increase in hepatocelluar
carcinomas only at 3.75 /ig/kg/day. DPH  was not carcinogenic
in B6C3F1 male mice.
      120 - 124 Cf K & K labs.
                              C-ll

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o
I
t->
to
                                                  TABLE 2


                             Carcinogenicity of 1,2-diphenylhydrazine in Mice*
Mf ^ Ir r- r> C^nl. r.
Sex f
50
50
male
50
50
50
50
female
47
50
Dose Treated Observed Hepatocellular
Post treatment Carcinomas
LD control
HD control

0.75 /ig/kg/day
3.75 jug/kg/day
LD control
HD control

0.375 /ig/kg/day
3.75 /ig/kg/day
0
0

78
78
0
0

78
78
95
96

17
17
96
96

17
18
12/50
6/48

11/47
8/46
2/47
1/50

4/39
20/43 p<0.001
Pulmonary
Carcinomas
5/50
5/49

1/47
0/46
2/46
3/50

3/38
2/40
        *Data taken from NCIf  1978


         LD = Low dose

         HD = High dose

         Dose = DPH in diet

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o
I
                                                        TABLE  3


                                    Carcinogenic!ty of 1,2-diphenylhydrazine in Rats
Sex I
SO
49
male
SO
SO
SO
50
female
50
50



Effects
	 Weeks 	  
Dose Treated Observed Hepatocellular Zymbal's Gland
Carcinoma
Ld control
HD control
5 ug/kg/day
18. B ug/kg/day
LD control
HO control
3 ug/kg/day
7.5 ug/kg/day
0
0
78
78
0
0
78
78
108
109
29
28
109
109
30
29
0/47
1/48
5/49 p 0.031
31/49 p 0.001
Hepatic Neoplastic
0/47
0/50
0/50
6/50 p 0.013
0/47
1/48
2/50
7/49 p 0.007
Nodules
0/48
0/50
1/50
0/50
Pulmonary
Carcinomas
1/47
1/48
3/49
1/48
Mammary carcinomas
1/48
0/50
3/50
6/50 p<0.013
    *Data taken from NCI, 1978.


     LD = Low dose
     HD = High dose

     Dose = % DPH in diet

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     In an additional study by Pliss in 1974, the carcino-
genic properties of DPH were studied over a period of 588
days in rats (N = 163) and C 57 mice (N = 110).  DPH was
suspended in sunflower seed oil and administered by s.c.
injection (40 mg/wk/rat and 5 mg/wk/mouse), and by addition
to food (30 mg/5 times/wk), or application to the skin
(30 mg/5 times/wk/rat and 2 mg/3 times/wk/mouse).
                           TABLE 4
                                                  *
           Carcinogenicity of DPH in Mice and Rats*
                                          Effects
     Species   Route   % Tumor Incidence          Tumors
     mice      s.c.  *       36.6
               PO           50

           epicutaneous     22.2
                              r habdomyosarcoma
                              pulmonary adenoma,
                              leukemia, liver
                              skin, lung, liver
     rats
s.c.
22.6
uterus, mammary,
Zymbal's gland, liver
spleen, lymphoid leukemia
     The data summarized in Table 4 indicate that
DPH produces a wide variety of tumors in both mice and rats.
 Data taken from Pliss, 1974.
                              C-14

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     In contrast to the NCI (1978)  study and the report
by Pliss (1974), Marhold, et al.  (1968)  and Spitz (1950)
did not find any significant increase in carcinogenicity
by DPH. These latter two studies  were difficult to interpret
due to the lack of specific information on the purity of
DPH, experimental design or statistical analysis.  The Pliss
study  (1974) should be used in a  cautious manner in indicting
DPH as a carcinogen.  The author  indicates 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 is given for  DPH treated animals, the
incidence of tumors in control animals is not presented
except in the case of the epicutaneous administration of
DPH.  Values of 17 percent vs. 22.2 percent for control
and DPH groups, respectively,  are presented but no statistical
analysis of these incidences is given.  The NCI  (1978) report
stands in marked contrast to the  other published studies
on the carcinogenecity of DPH.  It represents the only reli-
able study and indicates that DPH is carcinogenic.
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                    CRITERION FORMULATION
Existing Guidelines and Standards
     No existing guidelines or standards were found for
1,2-diphenylhydrazine.
Current Levels of Exposure
     No information is available on the concentration of
1,2-diphenylhydrazine in the atmosphere.
     1,2-Diphenylhydrazine has been found to be present
in drinking water at levels of 1 ;ug/l = 1 ppb (U.S. EPA,
1975).
     1,2-Diphenylhydrazine has not been found to be a natural
constituent of food.
Special Groups at Risk
     Manufacturers of dyes and Pharmaceuticals are subject
                                                      N
to occupational exposure.  Groups working in the laboratory
and forensic medicine may also be subject to 1,2-diphenyl-
hydrazine exposure.
Basis and Derivation of Criterion
     An evaluation of the subacute, acute and chronic toxicity,
with the exception of carcinogenicity is impossible because
of only scanty data.  No current guidelines or standards
presently exist for DPH. Diphenylhydrazine has been shown
to produce carcinogenic responses 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 criterion.
     More specifically, the data on the induction of cancer
in male and female rats and female mice were chosen for
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analysis because they all had significantly increased tumor
formation following DPH treatment (i.e. dietary).  The respec-
tive criterion levels obtained from applying the standard
water quality dose extrapolation/criteria calculation method-
ology is given in Table 5.
                                TABLE 5
      1,2-Diphenylhydrazine Induction of Tumors in Mice and Ratsa
Species     Sex                    Estimated Criterion Level
                                        at 10~3 Risk0
Mouse       Female                      1.43 /ig/1
Rat         Female, liver carcinoma     1.32 jug/1
                    mammary carcinoma   1.32 /ag/1
            Male, zymbol gland tumor    5.14 >ug/l
                  liver carcinoma       0.38/ig/l
*Data taken from NCI, 1978. (Tech No. 92, 1978)
 Calculated by applying a modified "one-hit" extrapolation
model described in the Federal Register 1062-5, 1979.
It can be seen that male rats appear to have the lowest
tolerance for DPH.
     Under the Consent Decree in NRDC vs. Train, criteria
are to state "recommended maximum permissible concentrations
(including where appropriate, zero) consistent with the
protection of aquatic organisms, human health, and recreational
activities."  DPH is suspected of being a human carcinogen.
Because there is no recognized safe concentration for human
carcinogens, the recommended concentration of DPH in water
for maximum protection of human health is zero.
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     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 concentrations of DPH corresponding to
several incremental lifetime cancer risk levels have been
estimated.  A cancer risk level provides 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"  indicates one additional
case of cancer for every million people exposed, and so
forth.
     In the Federal Register notice of availability of draft
ambient water quality criteria, EPA stated that it is consid-
ering setting criteria at an interim target risk level of
10~5, 10"6 or 10~7 as shown in the table below.
Exposure Assumptions               Risk Levels and Corresponding Criteria
                                   0    10~7        10"6      IGf5
2 liters of drinking water         0     4 ng/1      40 ng/1   400 ng/1
and consumption of 18.7
grams fish and shellfish (2)
Consumption of fish and            0    .019 jug/1  0.19 jug   1.9 >ug/l
shellfish only.
(1)  Calculated by applying a modified "one-hit" extrapolation
     model described in the PR 15926, 1979. Appropriate
     bioassay data used in the calculation of the model
     are presented in Appendix I.  Since the extrapolation
     model is linear to low doses, the additional lifetime
     risk is directly proportional to the water concentration.
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     Therefore,  water concentrations corresponding to other
     risk levels can be derived by multiplying or dividing
     one of the  risk levels and corresponding water concen-
     trations shown in the table by factors such as 10,
     100, 1000 and so forth.
(2)  Twenty-one  percent of the DPH exposure results from
     the consumption of aquatic organisms which exhibit
     an average  bioconcentration potential of 29 fold.
     The remaining percent of DPH exposure results from
     drinking water.
     Concentration levels were derived assuming a lifetime
exposure to various amount 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 corresponding DPH concen-
trations.
    -. Although a total exposure evaluation for DPH is desirable
                    i
there is no data to support a total exposure analysis.
The criteria presented, therefore, assume an incremental
risk from assumed ambient water exposure only.
     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, there is no information on other effects
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.
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                          REFERENCES
Anthony, A.M., et al. 1970. Tumors of the urinary bladder:
an analysis of the occupations of 1030 patients in Leeds,
England.  Jour. Natl. Can. Inst. 45:  879.

CRC. 1978. Handbook of chemistry and physics. 59th ed. CRC
Press. West Palm Beach, Fla.

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.

Haley, T.J. 1975. Benzidine revisited: a review of the litera-
ture and problems associated with the use of benzidine and
its congeners. Clin. Tox. 8: 13.

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

International Agency for Research on Cancer. 1974. Hydrazine
and its derivatives.  Monog. on the evaluation of carcinogenic
risk of chemicals to man.  4: 81.
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Kenner,  J. 1968. Benzidine rearrangement. Nature. 219: 153.

Kulyanskii, et al. 1976.  Kirk-Othmer Encyclopedia of Chemical
Technology. 1963. 2nd ed. Vol.  3. New York: Interscience.
p. 408.

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

NCI Publication NO.  (NIH) 78-1342. 1978. Bioassay of hydrazo-
benzene for possible carcinogenicity.

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

Registry of Toxic Effects of Chemical Substances.  II: 1977.
         i
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.

Stanford Research Institute. 1977. 1977 Directory of chemical
producers, U.S.A. Menlo Park, Calif.

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Button, W.L. 1967. Heterocyclic and miscellaneous nitrogen
compounds.  Industrial Hygiene and Toxicology. Vol. II:
2171. Toxicology, 2nd ed. F.A. Patty, ed. New York: Interscierice
Publishers.

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

Wenner, W. 1967. Malonic acid and derivatives in Kirk-Othmer
Encyclopedia of Chemical Technology. 2nd ed. Vol. 12: 857.
New York: Interscience Publishers.

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

Wynder, E.L., et al. 1963. An epidemiological investigation
of cancer in the bladder. Cancer 16: 1388.
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                         APPENDIX I

            Summary and Conclusions  Regarding the
         Carcinogenicity of 1,2,-Diphenylhydrazine*

     I,2-Diphenylhydrazine is  used primarily in dye manu-

facturing 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 administra-

tion.  Male rats, receiving dietary concentrations of 0.03

percent 1,2-diphenylhydrazine, developed hepatocellular

carcinomas and squamous cell carcinomas of the Zymbal glands.

Female rats, receiving 0.01 percent  1,2-diphenylhydrazine

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

carcinomas.

     The carcinogenic responses induced in male and female

rats and female mice constitute substantial evidence that

1,2-diphenylhydrazine 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 concentration of 0.03 percent (300

ppm)  1,2-diphenylhydrazine 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   is

0.40 micrograms per liter.

*This summary has been prepared and  approved by the Carcinogens
 Assessment Group of EPA on June 15, 1979.
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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 nodules in male Fischer 344 rats, exposed to
0.03 percent (300 ppm) 1,2-diphenylhydrazine in the diet
ad_ libitum for 78 weeks  (NCI, 1978). The incidence of hepato-
cellular carcinomas and neoplastic nodules was 37/49 and
1/48 in the treated and control groups, respectively. The
criterion was calculated from the following parameters:
     nt =37                      d* = 15 mg/kg/day
     Nt = 49                       F = .0187 kg/day
     nc = 1                        R = 29
     Nc = 48                       W = 0.375 kg
     Le = 104 weeks
     le =  78 weeks
     L  = 104 weeks
Based on these parameters, the "one-hit" slope (BH) is 0.715
(mg/kg/day)  .  The resulting water concentration of 1^2-
diphenylhydrazine, calculated to keep the individual lifetime
cancer risk below 10" , is 0.40 >ig/l.
*    The dose  (expressed as mg/kg  (body weight) /day) is
based on the assumption that the amount of diet consumed
by rats each day was five percent of their body weights0
     0.05 x 0.375 kg = 0.01875 kg diet/day
     0.01875 kg diet/day x 300 mg/kg = 5.625 mg 1,2 DPH/day
     5.625 mg 1,2 DPH/day/0.375 kg = 15 mg/kg/day
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