,,.„«, FINAL DRAFT
United States ccnn riu rni*
Env.ronmental Protect.on ECAO-C1N-G034
Agency May, 1988
Research and
Development
HEALTH AND ENVIRONMENTAL EFFECTS DOCUMENT
FOR 3,3'-DICHLOROBENZIDINE
Prepared for
OFFICE OF SOLID WASTE AND
EMERGENCY RESPONSE
Prepared by
Environmental Criteria and Assessment Office
Office of Health and Environmental Assessment
U.S. Environmental Protection Agency
Cincinnati, OH 45268
DRAFT: 00 NOT CITE OR QUOTE
NOTICE
This document Is a preliminary draft. It has not been formally released
by the U.S. Environmental Protection Agency and should not at this stage be
construed to represent Agency policy. It Is being-circulated for comments
on Its technical accuracy and oollcy Implications.
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DISCLAIMER
This report Is an external draft for review purposes only and does not
constitute Agency policy. Mention of trade names or commercial products
does not constitute endorsement or recommendation for use.
11
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PREFACE
Health and Environmental Effects Documents (HEEOs) are prepared for the
Office of Solid Waste and Emergency Response (OSWER). This document series
Is Intended to support listings under the Resource Conservation and Recovery
Act (RCRA) as well as to provide health-related limits and goals for emer-
gency and remedial actions under the Comprehensive Environmental Response,
Compensation and Liability Act (CERCLA). Both published literature and
Information obtained for Agency Program Office files are evaluated as they
pertain to potential human health, aquatic life and environmental effects of
hazardous waste constituents. The literature searched for In this document
and the dates searched are Included In "Appendix: Literature Searched."
Literature search material 1s current up to 8 months previous to the final
draft date listed on the front cover. Final draft document dates (front
cover) reflect the date the document Is sent to the Program Officer (OSWER).
Several quantitative estimates are presented provided sufficient data
are available. For systemic toxicants, these Include Reference doses (RfDs)
for chronic and subchronlc exposures for both the Inhalation and oral
exposures. The subchronlc or partial lifetime RfD, 1s an estimate of an
exposure level that would not be expected to cause adverse effects when
exposure occurs during a limited time Interval I.e., for an Interval that
does not constitute a significant portion of the llfespan. This type of
exposure estimate has not been extensively used, or rigorously defined as
previous risk assessment efforts have focused primarily on lifetime exposure
scenarios. Animal data used for subchronlc estimates generally reflect
exposure durations of 30-90 days. The general methodology for estimating
subchronlc RfDs Is the same as traditionally employed for chronic estimates,
except that subchronlc data are utilized when available.
In the case of suspected carcinogens, RfDs are not estimated. Instead,
a carcinogenic potency factor, or q-j* (U.S. EPA, 1980) 1s provided. These
potency estimates are derived for both oral and Inhalation exposures where
possible. In addition, unit risk estimates for air and drinking water are
presented based on Inhalation and oral data, respectively.
Reportable quantities (RQs) based on both chronic toxldty and cardno-
genlclty are derived. The RQ Is used to determine the quantity of a hazard-
ous substance for which notification 1s required In the event of a release
as specified under the Comprehensive Environmental Response, Compensation
and Liability Act (CERCLA). These two RQs (chronic toxldty and carclno-
genldty) represent two of six scores developed (the remaining four reflect
1gn1tab1l1ty, reactivity, aquatic toxldty, and acute mammalian toxldty).
Chemical-specific RQs reflect the lowest of these six primary criteria. The
methodology for chronic toxldty and cancer based RQs are defined In U.S.
EPA, 1984 and 1986a, respectively.
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EXECUTIVE SUMMARY
3,3'-01chlorobenz1d1ne (CAS number 91-94-1) 1s a gray to purple crystal-
line solid at room temperature (Hawley, 1981). It Is available commercially
1n the form of Us dlhydrochlorlde salt (IARC, 1982). Commercial production
Involves alkaline reduction of o-chloronltrobenzene, followed by rearrange-
ment with hydrochloric acid to form 3,3'-d1chlorobenz1d1ne dlhydrochlorlde
(IARC, 1982). Bofors Nobel, Inc. In Muskegon, MI, and The Upjohn Co. 1n
North Haven, CT, are currently the only U.S. manufacturers of 3,3'-d1chloro-
benzldlne dlhydrochlorlde (SRI, 1987). 3,3'-D1chlorobenz1d1ne dlhydro-
chlorlde 1s Imported Into the United States; during 1983, 1.104 million
pounds of 3,3'-d1chlorobenz1d1ne base and salts was Imported through the
principal U.S. customs districts (USITC, 1984). Essentially 100% of all
3,3'-d1chlorobenz1d1ne (1n the form of Its dlhydrochlorlde) consumed In the
United States 1s used as an Intermediate for organic pigments (IARC, 1982;
HSD8, 1987).
In the atmosphere, 3,3'-d1chlorobenz1d1ne 1s expected to exist primarily
1n the partlculate form. 3,3'-D1chlorobenz1d1ne, In both vapor and partlcu-
late form, 1s expected to undergo rapid photolysis In the atmosphere.
Reaction of 3,3'-d1chlorobenz1d1ne vapor with photochemlcally generated
hydroxyl radicals (t, ..=10 hours) (Atkinson, 1987) may be a minor removal
mechanism. In water, this compound would undergo rapid photodegradatlon
(t 2=90 sec) 1n the surface layers of water (Banerjee et al., 1978).
This compound photodegrades to monochlorobenzldlne, benzldlne and a number
of brightly colored water-Insoluble compounds (Banerjee et al., 1978).
Beyond the reach of light penetration, this compound would rapidly adsorb
to sediment and partlculate matter where 1t Is tightly bound. Adsorption 1s
1v
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expected to proceed Initially by a rapid, but reversible, physical adsorp-
tion process followed by much slower Irreversible covalent bonding (Appleton
et al., 1978; S1kka et al., 1978). It has also been speculated that
3,3'-d1chlorobenz1d1ne may be oxidized by naturally occurring cations (e.g.,
Fe* ) found In sediments (Callahan et al., 1979). Rapid uptake and
bloaccumulatlon 1n aquatic organisms Is also expected to occur (Appleton and
Slkka, 1980; Freltag et al., 1985). Volatilization, m1crob1al degradation
and chemical hydrolysis are not expected to be Important fate processes 1n
water. In soil, 3,3'-d1chlorobenz1d1ne Is expected to adsorb tightly to
soil, and over time, Irreversibly bind with humates 1n the soil (Boyd et
al., 1984). It has also been speculated that oxidation by reaction with
cattonlc constituents of soil (e.g., Fe* , Cu* ) may also occur
(Callahan et al.. 1979; Dem1rj1an et al., 1987). If exposed to sunlight on
soil surfaces, 3,3'-d1chlorobenz1d1ne Is expected to photodegra.de rapidly.
Volatilization and mlcroblal degradation are not expected to be significant
fate processes (Callahan et al., 1979).
Exposure to 3,3'-d1chlorobenz1d1ne Is most likely to occur In occupa-
tional settings, particularly where this compound 1s manufactured or where
3,3'-d1chlorobenz1d1ne-based dyes are manufactured or used (HSDB, 1987).
The general public may be exposed to low levels of 3,3'-d1chlorobenz1d1ne
during use of paints, pigments or enamels derived from this compound. The
most probable routes of exposure are Inhalation of dusts or mists containing
this compound and dermal contact {HSDB, 1987). 3,3'-D1chlorobenz1d1ne has
been detected 1n samples of surface water, sediment, fish and Industrial
effluent (U.S. EPA, 1981, 1987b; IARC, 1982; Hauser and Bromberg, 1982).
This compound has not been detected 1n urban runoff samples collected as
part of the U.S. EPA Nationwide Urban Runoff Program (Cole et al., 1984).
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The U.S. EPA STORET Data Base Indicates that the mean concentrations of
3,3'-d1chlorobenz1d1ne In whole water, sediment (wet and dry weight basis)
and fish tissue samples collected throughout the United States are 46
vg/l, 3222 yg/kg dry wt., 0.026 and 5.8 mg/kg wet wt., respectively
(U.S. EPA, 1987b).
There are few data regarding the aquatic toxlclty of 3,3'-d1chloro-
benzldlne. A study by Appleton et al. (1978) Indicated that In bluegllls,
Lepomls macrochlrus. toxic levels of 3,3'-d1chlorobenz1d1ne accumulated
before a 3,3'-d1chlorobenz1d1ne equilibrium was reached between water and
fish. There were many mortalities when the whole body residues of fish
exposed to l4C-3,3'-d1chlorobenz1d1ne exceeded 150 ppm.
The absorption of 3,3'-d1chlorobenz1d1ne following administration of the
compound by relevant routes (I.e., Inhalation or oral exposure) has not been
studied extensively. Hsu and S1kka (1982) reported that 3,3'-d1chlorobenz1-
dlne 1s rapidly and extensively absorbed following oral administration of
the compound to rats. Melgs et al. (1954) and Susklnd (1983) reported that
the skin Is the most significant route of entry of 3,3'-d1chlorobenz1d1ne
Into the body 1n cases of occupational exposure. The half-life of dis-
appearance of a topically applied 3,3'-d1chlorobenz1d1ne dose from the
shaved backs of rats was determined to be 24.1 hours (Shah and GuthMe,
1983).
Following a single oral dose of radlolabled 3,3'-d1chlorobenz1d1ne to
rats, the principal organs In which radioactivity was found were the liver,
kidneys, lungs and spleens (Hsu and S1kka, 1982). Multiple oral
3,3'-d1chlorobenz1d1ne dosing led to tissue levels of radioactivity 3-4
times higher than the levels observed following a single oral dose, but .a
multiple dosing schedule did not result 1n substantial retention of
radioactivity (Hsu and S1kka, 1982).
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The extent of metabolism and the pathways of 3,3'-d1chlorobenz1d1ne
metabolism are not clear. Ring hydroxylatlon products of 3,3'-dlchloro-
benzldlne were not found 1n the urine of humans and dogs given an oral
3,3'-d1chlorobenz1d1ne dose (Troll, n.d.), and Shrlner et al. (1978)
suggested that chloMnatlon of benzldlne blocks ring hydroxylatlon reactions
of 3,3'-d1chlorobenz1d1ne for both electronic and sterlc reasons. Hsu and .
Slkka (1982), however, provided evidence that 3,3'-d1chlorobenz1d1ne Is
metabolized extensively In rats.
Several possible metabolites of 3,3'-d1chlorobenz1d1ne, tentatively
Identified by chromotographlc procedures, Include mono-N-acetyl 3,3'-dl-
chlorobenzldlne In the urine of monkeys (Kellner et al., 1973) and benzldlne
and some possible glucuronlde conjugates 1n the urine of rats (Aksamltnala,
1959).
Elimination of both radlolabeled 3,3'-dlchlorobenzldlne and total radio-
activity from the plasma of orally dosed rats was blphaslc showing an
Initial rapid decline followed by a slower disappearance phase (Hsu and
Slkka, 1982). Similarly, blphaslc elimination of total radioactivity was
observed In the principal organs of distribution (I.e., liver, lung and
kidney and spleen) (Hsu and Slkka, 1982).
The bile appears to be a significant route of excretion for both .
3,3'-d1chlorobenz1d1ne and Its metabolites. Experiments with rats Indicated
that ~90% of the administered radioactivity 1s excreted In the urine and
feces following an oral dose of 3,3'-dlchlorobenzldlne (Hsu and Slkka,
1982). Approximately 6554 of the administered radioactivity was excreted In
the feces and the major source of the radioactivity found In the feces
originated from the bile. HepatoblHary excretion of 3,3'-dlchlorobenzldlne
and Us metabolites also occurred 1n rhesus monkeys (Kellner et al., 1973).
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The fecal route appears to be the most significant route of elimination
of 3,3'-d1chlorobenz1d1ne and metabolites In humans (Troll, n.d.); 3,3'-d1-
chlorobenzldlne has also been reported to be present In the urine of occupa-
tlonally-exposed Individuals (Helgs et al., 1954; London and Bolano, 1986;
Slngal and Lee, 1985).
Pertinent data regarding the systemic toxlclty of 3,3'-d1chlorobenz1d1ne
following either subchronlc or chronic Inhalation exposure In humans and
animals were not located In the available literature. Stula et al. (1978)
Indicated that beagle dogs exposed to 3,3'-d1chlorobenz1d1ne orally for
periods up to 7.1 years showed signs of liver toxlclty In the form of
elevated SGPT activities. The oral L05Q of 3,3'-d1chlorobenz1d1ne 1n rats
has been reported to be -4 and 7 g/kg bw for dlhydrochlorlde salt and
3,3-d1chlorobenz1d1ne, respectively (ACGIH, 1986).
Pertinent data regarding the carclnogenlcHy of 3,3'-d1chlorobenz1d1ne
following Inhalation exposure In humans and animals were not located In the
available literature cited In Appendix A. Oral administration of 3,3'-d1-
chlorobenzldlne has been shown to produce a variety of tumors 1n rats
(PUss, 1959; Stula et al., 1975; GHswold et al., 1968), urinary bladder
and liver tumors 1n dogs (Stula et al., 1978) and hamsters (Sellakumar et
al., 1969), and hepatomas 1n mice (Osanal, 1976). Subcutaneous administra-
tion of 3,3'-d1chlorobenz1d1ne has also been demonstrated to produce tumors
1n rats (PUss, 1959, 1963).
Both additive and synerglstlc tumorlgenie effects were noted In rats
following simultaneous or sequential administration of low levels of
3,3'-d1chlorobenz1d1ne along with low levels of other carcinogens (I.e., BBN
alone or BBN, FANFT and 2-AAF sequentially (Ito et-al., 1983).
3,3'-D1chlorobenz1d1ne has also been demonstrated to function as a
transplacental carcinogen 1n mice (Golub et al., 1975).
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3,3'-D1ch1orobenz1d1ne has been suspected of being a human carcinogen
because of Us carcinogenic effects 1n animals and because H resembles
benzldlne, a known human bladder carcinogen. Evidence from three
ep1dem1olog1cal studies {Gerarde and Gerarde, 1974; Maclntyre, 1975; Gadlan,
1975), however. Is Inadequate to suggest that 3,3'-d1chlorobenz1d1ne 1s a
bladder carcinogen In humans.
3,3'-D1chlorobenz1d1ne has been demonstrated to be mutagenlc towards
Salmonella typhlmurlum 1n the Ames assay, both with and without metabolic
activation. Metabolic activation (I.e., presence of liver S-9) has, however,
been demonstrated to Increase the mutagenlclty of 3,3'-d1chlorobenz1d1ne
from 3 to 50-fold (Garner, 1975; Lazear and Louie, 1977; DeFrance et al.,
1986). 3,3'-D1chlorobenz1d1ne has also been demonstrated to be active In 1.n_
vitro assays measuring unscheduled DNA synthesis (Martin et al., 1978) and
sister chromat1d exchange (Shlralshl, 1986).
Pertinent data regarding the tetratogenlcHy of 3,3'-d1chlorobenz1d1ne
were not located In the available literature. One study (Shabad et al.,
1972) demonstrated that transplacental exposure of mice to 3,3'-d1chloro-
benzldlne had effects on the growth of embryonic kidney cells In culture.
Because 3,3'-d1chlorobenz1d1ne has been demonstrated to be a carcinogen
In animals, 1t 1s placed In EPA Group 2B, a probable human carcinogen.
Therefore, Inhalation and oral RfDs were not derived. A lack of pertinent
data on the carclnogenlcHy of 3,3'-d1chlorobenz1d1ne following Inhalation
exposure precluded the derivation of an Inhalation q,*. An oral q,* of
1.2 (mg/kg/day)"1 was derived for humans from the study of Stula et al.
(1975) In which female rats fed 3,3'-d1chlorobenz1d1ne 1n the diet (1000
ppm) over the course of a lifetime had a statistically significant Increased
Incidence of mammary adenocardnomas compared with controls. The levels of
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3,3'-d1chlorobenz1d1ne In drinking water associated with Increased lifetime
risk at risk levels of 10~5, 10~« and 10~7 are 3xlO"4, 3xlO~s and
3xlO~* mg/a, respectively. An F factor of 8.4 (mg/kg/day)"1, placing
3,3'-d1chlorobenz1d1ne 1n Potency Group 2, was also derived. Because
3,3'-d1chlorobenz1d1ne Is categorized In EPA Group B2 and Potency Group 2,
the compound has a MEDIUM hazard ranking under CERCLA. A medium hazard
ranking 1s associated with an RQ of 10. An RQ based on systemic toxlclty
was also derived from a study by Stula et al. (1978) 1n which dogs given an
oral dose of 3,3'-d1chlorobenz1d1ne (100 mg/day) over an extended period of
time (I.e., up to 7.1 years) showed signs of liver toxldty 1n the form of
elevated SGPT activities. This effect (liver toxlclty) and this dose are
associated with an RQ of 1000. Because 3,3'-d1chlorobenz1d1ne has been
demonstrated to be carcinogenic 1n animals, however, the more conservative
RQ of 10 derived from the carclnogenlclty data Is recommended.
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TABLE OF CONTENTS
Page
1. INTRODUCTION 1
1.1. STRUCTURE AND CAS NUMBER 1
1.2. PHYSICAL AND CHEMICAL PROPERTIES 1
1.3. PRODUCTION DATA 2
1.4. USE DATA 2
1.5. SUMMARY 3
2. ENVIRONMENTAL FATE AND TRANSPORT 4
2.1. AIR : 4
2.1.1. Reaction with Hydroxyl Radicals 4
2.1.2. Reaction with Ozone 4
2.1.3. Photolysis 4
2.2. WATER 5
2.2.1. Hydrolysis 5
2.2.2. Oxidation 5
2.2.3. Photolysis 5
2.2.4. M1crob1al Degradation 6
2.2.5. Volatilization 6
2.2.6. B1oaccumulat1on 7
2.2.7. Adsorption 7
2.3. SOIL 8
2.3.1. Hydrolysis 8
2.3.2. Oxidation 8
2.3.3. Photolysis 8
2.3.4. M1crob1al Degradation 8
2.3.5. Adsorption 8
2.3.6. Volatilization 9
2.4. SUMMARY 9
3. EXPOSURE 11
3.1. WATER 11
3.2. FOOD 11
3.3. INHALATION 11
3.4. DERMAL 14
3.5. SUMMARY 14
4. AQUATIC TOXICITY 15
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TABLE OF CONTENTS (cont.)
Page
5. PHARMACOKINETCS 16
5.1. ABSORPTION 16
5.2. DISTRIBUTION 17
5.3. METABOLISM 18
5.4. EXCRETION 19
5.5. SUMMARY 21
6. EFFECTS 23
6.1. SYSTEMIC TOXICITY. . . . '. 24
6.1.1. Inhalation Exposures 24
6.1.2. Oral Exposures 24
6.1.3. Other Relevant Information 24
6.2. CARCINOGENICITY 25
6.2.1. Inhalation 25
6.2.2. Oral 25
6.2.3. Other Relevant Information 33
6.3. MUTAGENICITY 35
6.4. TERATOGENICITY 38
6.5. OTHER REPRODUCTIVE EFFECTS 38
6.6. SUMMARY 39
7. EXISTING GUIDELINES AND STANDARDS 41
7.1. HUMAN 41
7.2. AQUATIC 41
8. RISK ASSESSMENT 42
8.1. CARCINOGENICITY 42
8.1.1. Inhalation 42
8.1.2. Oral 42
8.1.3. Other Routes 43
8.1.4. Weight of Evidence 43
8.1.5. Quantitative Risk Estimates 44
8.2. SYSTEMIC TOXICITY 45
8.2.1. Inhalation Exposures 45
8.2.2. Oral Exposures 46
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TABLE OF CONTENTS (cont.)
Page
9. REPORTABLE QUANTITIES 47
9.1. BASED ON SYSTEMIC TOXICITY 47
9.2. BASED ON CARCINOGENICITY 51
10. REFERENCES 54
APPENDIX A: LITERATURE SEARCHED 66
APPENDIX B: CANCER DATA SHEET FOR DERIVATION OF q]* 69
APPENDIX C: SUMMARY TABLE FOR 3,3'-DICHLOROBENZIDINE 70
X111
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LIST OF TABLES
_No^_ Title Page
3-1 Monitoring Data for 3,3'-D1chlorobenz1d1ne In Various Media . 12
6-1 Incidence of Tumors In Female Beagle Dogs Given
3,3'-D1chlorobenz1d1ne (-100% pure) Orally by Capsule .... 27
6-2 Incidence of Tumors 1n Chr-CD Rats Treated With
3,3'-D1chlorobenz1d1ne 1n the Diet 29
6-3 Incidence of Hepatomas In Male ICR/JCL Mice Fed 0.1%
3,3'-D1chlorobenz1d1ne In the Diet 31
6-4 Mutagenlclty Testing of 3,3'-D1chlorobenz1d1ne 36
9-1 Oral Toxlclty Summary for 3,3'-D1ch'lorobenz1d1ne Using
the Female Beagle 48
9-2 Oral Composite Scores for 3,3'-D1chlorobenz1d1ne Using
the Dog 49
9-3 3,3'-D1chlorobenz1d1ne: Minimum Effective Dose (MED) and
Reportable Quantity (RQ) 50
9-4 Derivation of Potency Factor (F) for 3,3'-D1chlorobenz1d1ne . 53
xlv
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LIST OF ABBREVIATIONS
2-AAF 2-Acetylam1nofluorene
BCF Bloconcentratlon factor
BBN N-butyl-N-(4-hydroxy butyl) nltrosamlne
bw Body weight
CAS Chemical Abstract Service
CS Composite score
DHBA 9,lO-D1methyl-l,2-benzanthracene
DMSO Dimethyl sulfoxlde
DNA Deoxyrlbonuclelc add
FANIFT N-(4-(5-n1tro-2-furyl )-2-th1azolyl )formam1de
HPLC High performance liquid chromatography
Koc Soil sorptlon coefficient standardized
with respect to organic carbon
K Octanol/water partition coefficient
ow
LC«io Concentration lethal to 50% of recipients
(and all other subscripted concentration levels)
LD5Q Dose lethal to 50% of recipients
MED Minimum effective dose
ppb Parts per billion
ppm Parts per million
RfD Reference dose
RQ Reportable quantity
RV. Dose-rating value
RV Effect-rating value
SGPT Serum glutamlc pyruvlc transamlnase
TLV Threshold limit value
TWA Time-weighted average
UV Ultraviolet
w/w Weight per weight
xv
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1. INTRODUCTION
1.1. STRUCTURE AND CAS NUMBER
3,3'-D1chlorobenz1d1ne Is also known as DCB, C.I. 23060; 4,4'-d1am1no-
3,3,'-d1chlorob1phenyl; and 3,3'-d1chloro-(l,V-b1phenyl )-4,4'-d1am1ne (IARC,
1982). The structure, CAS Registry number, empirical formula and molecular
weight are as follows:
Cl
CAS Registry number: 91-94-1
Empirical formula: c12HioC12N2
Molecular weight: 253.1
1.2. PHYSICAL AND CHEMICAL PROPERTIES
3,3'-D1chlorobenz1d1ne Is a gray to purple crystalline solid at room
temperature (Hawley, 1981). The reactions of this compound are similar to
those of benzldlne and other benzldlne derivatives, e.g., formation of
dlazonlum salts and acyl and alky! derivatives (IARC, 1982; Ferber, 1978).
3,3'-D1chlorobenz1d1ne 1s readily soluble In benzene, dlethyl ether, ethanol
and glacial acetic add but almost Insoluble 1n water (IARC, 1982). Selected
physical properties are as follows:
Melting point:
Boiling point:
Vapor pressure at 25°C:
Water solubility at 25°C:
Log Kow:
133°C
422°C
(estimated)
4.2xlO~7 mm Hg
(estimation based on
the formula:
TM = 0.5839 Tb)
3 mg/l
3.51
Ferber, 1978
Neely and Blau, 1985
Neely and Blau, 1985
Banerjee et al., 1980
Hansch and Leo, 1985
0087d
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03/31/88
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Conversion factor 1n air: 1 ppm = 0.0966 mg/m3 IARC, 1982
1.3. PRODUCTION DATA
3,3'-D1chlorobenz1d1ne 1s produced commercially by alkaline reduction of
o-chloronltrobenzene followed by rearrangement of the resulting hydrazo
compound with hydrochloric acid to form the dlhydrochloHde salt (IARC,
1982). The dlhydrochloMde salt 1s the commercially available form of
3,3'd1chlorobenz1d1ne (IARC, 1982). The U.S. EPA TSCA Production File (U.S.
EPA, 1977) contained no production data on 3,3'-d1chlorobenz1d1ne, but
contained the following Information on 3,3'-d1chlorobenz1d1ne
dlhydrochloMde:
Company/Location 1977 Production/Import Volume
The Upjohn Co., confidential
North Haven, CT
Sun Chemical Corp., confidential
Cincinnati, OH (Importer)
Bofors Lakeway Inc., 1-10 million pounds
Huskegon, HI
SRI (1987) listed Bofors Nobel, Inc. 1n Muskegon, MI, and The Upjohn Co. 1n
North Haven, CT, as the only current domestic manufacturers of 3,3'-d1-
chlorobenzldlne dlhydrochlorlde. Domestic production volume data for recent
years were not located 1n the available literature cited In Appendix A.
During 1983, 1.104 million pounds of 3,3'-d1chlorobenz1d1ne base and salts
was Imported through principal U.S. custom districts (USITC, 1984).
1.4. USE DATA
Essentially 100X of all 3,3'-d1chlorobenz1d1ne (1n the form of
dlhydrochlorlde salt) consumed 1n the United States 1s used as an
Intermediate for organic pigments (IARC, 1982; HSDB, 1987). At least 95
tetrazo dyes can be derived from 3,3'-d1chlorobenz1d1ne, but only 5
0087d -2- 03/31/88
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(Pigments Orange 13 and 14 and Pigments Yellow 12, 13 and 14) are currently
used 1n the United States (Boyd et al., 1984; ShMner et al., 1978). These
dyes are used 1n various paints, enamels and lacquers (Shrlner et al.,
1978). 3,3'-D1chloro- benzldlne was also used In color tests for the
presence of gold and used alone, or 1n blends with 4,4'-methyleneb1s-
(2-chloroan111ne) as a curing agent for liquid castable polyurethane
elastomers (IARC, 1982).
1.5. SUMMARY
3,3'-01chlorobenz1d1ne (CAS number 91-94-1) Is a gray to purple crystal-
line solid at room temperature (Hawley, 1981). It Is available commercially
1n the form of Us dlhydrochlorlde salt (IARC, 1982). Commercial production
Involves alkaline reduction of o-nUrochlorobenzene, followed by rearrange-
ment with hydrochloric add to form 3,3'-d1chlorobenz1d1ne dlhydrochlorlde
(Ferber, 1978; IARC, 1982). Bofors Nobel, Inc. In Huskegon, Ml, and. The
Upjohn Co. In North Haven, CT, are currently the only U.S. manufacturers of
3,3'-d1chlorobenz1d1ne dlhydrochlorlde (SRI, 1987). 3,3'-D1chlorobenz1d1ne
dlhydrochlorlde Is Imported Into the United States; during 1983, 1.104
million pounds of 3,3'-d1chlorobenz1d1ne base and salts was Imported through
the principal U.S. customs districts (USITC, 1984). Essentially 100% of all
3,3'-d1ch1orobenz1d1ne (In the form of Us dlhydrochlorlde salt) consumed In
the United States 1s used as an Intermediate for organic pigments (IARC,
1982; HSDB, 1987).
0087d -3- 03/31/88
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2. ENVIRONMENTAL FATE AND TRANSPORT
2.1. AIR
Based on a vapor pressure of 4.2xlO~7 mm Hg at 25°C, 3,3'-d1chloroben-
zldlne 1s expected to exist primarily In participate form 1n the atmosphere.
2.1.1. Reaction with Hydroxy Radicals. The rate constant for the
reaction of 3,3'-d1chlorobenz1d1ne vapor with photochemically generated
hydroxyl radicals In the atmosphere was estimated to be ~4xlO"11
cmVmolecules-sec at 25°C using the method of Atkinson (1987). Assuming
that the average ambient hydroxyl radical concentration Is SxlO9 mole-
cules/cm3, the hydroxyl reaction half-life for 3,3'-d1chlorobenz1d1ne
vapor was estimated to be -10 hours. Since only small amounts of 3,3'-d1-
chlorobenzldlne released to the atmosphere are expected to exist In the
vapor phase, the environmental significance of this reaction would be
limited.
2.1.2. Reaction with Ozone. 3,3'-01chlorobenz1d1ne Is not susceptible to
oxidation by reaction with ozone molecules 1n the atmosphere (U.S. EPA,
1987a).
2.1.3. Photolysis. 3,3'-D1chlorobenz1d1ne In methanol, ethanol or water
exhibits strong absorption of UV light wavelength 1n the environmentally
significant range (wavelength 290-340nm) (Sadtler, n.d.; Callahan et al..
1979). 3,3'-D1chlorobenz1d1ne, adsorbed onto silica gel, underwent 41.2%
degradation (based on volatile compounds and C0_ evolved 1n the
photodegradatlon process) when Irradiated with light (wavelengths >290 nm)
for 17 hours (Freltag et al., 1985). This Information combined with the
observed rapid photolysis of 3,3'-d1chlorobenz1d1ne In water suggests that
3,3'-d1chlorobenz1d1ne 1n both partlculate and vapor form would rapidly
photolyze In the atmosphere.
0087d -4- 04/01/88
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2.2. WATER
2.2.1. Hydrolysis. Chemical hydrolysis of 3,3'-d1chlorobenz1d1ne In
water 1s not expected to be environmentally significant (Mabey et al., 1981;
U.S. EPA, 1981).
2.2.2. Oxidation. Pertinent data regarding the oxidation of 3,3'-d1-
chlorobenzldlne In natural waters were not located 1n the available litera-
ture cited 1n Appendix A. UnsubstUuted benzldlne 1s rapidly oxidized by
Iron (f3) and several other naturally occurring cations that can be found In
environmental waters as solvated cations, as complexes of humlc acids and as
structural components of mlcrocrystalUne clays. Whether or not 3,3'-d1-
chlorobenzldlne would be oxidized rapidly In a similar manner depends upon
Us 1on1zat1on potential 1n relation to the lonlzatlon potential of
benzldlne Itself. Because of the two chlorine substltuents on the aromatic
rings of 3,3'-d1chlorobenz1d1ne, this compound would have a lesser tendency
to lose an electron ( I.e., to oxidize) than unsubstUuted benzldlne.
Nevertheless, 1t has been speculated that oxidation by metal cations and
other environmental electron acceptors may contribute to the degradation of
3,3'-d1chlorobenz1d1ne 1n sediments (Callahan et al., 1979).
2.2.3. Photolysis. Dilute (10~5 M) aqueous solutions of 3,3'-d1chloro-
benzldlne 1n quartz tubes at neutral pH were exposed to noonday summer
sunlight at Syracuse, NY. Irradiation resulted 1n a half-life of -90 sec
(Banerjee et al., 1978). 3,3'-D1chlorobenz1d1ne photodegraded to mono-
chlorobenzldlne, benzldlne and a number of brightly colored water-Insoluble
materials. The same Intermediate products were formed upon photolysis of
aqueous solutions under acidic conditions and aqueous solutions treated with
chlorine water (Banerjee et al., 1978). The photolysis half-life for
3t3'-d1chlorobenz1d1ne In organic solvents Is markedly longer than 1n
aqueous solutions, and the mechanism of dechlorlnatlon does not appear to
0087d -5- 03/31/88
-------
Involve simple carbon-chlorine bond homolysls. Reduced photo-reactivity In
nonaqueous solutions, along with the high K of 3,3'-d1chlorobenz1d1ne,
might lead to enhanced stability of this compound In water contaminated with
hydrocarbons (Banerjee et al., 1978).
2.2.4. M1crob1al Degradation. [l4C]-3,3'-d1chlorobenz1d1ne was Incu-
bated (21°C 1n the dark) In natural water samples obtained from a eutrophlc
lake and a mesotrophlc lake. After a 1-month Incubation period, 75% of the
original 3,3'-d1chloroben2ld1ne was detected when assayed by HPLC and no
metabolites were detected (Appleton et al., 1978; Slkka et al., 1978).
Therefore, 3-3'-d1chlorobenz1d1ne appears to resist blodegradatlon by
aquatic microbes. Slkka et al. (1978) speculated that loss of the compound
was primarily the result of adsorption to sediments or accumulation by
aquatic organisms. [The BCF for 3,3'-d1chlorobenz1d1ne In activated sludge
Is 3100 (Freltag et al., 1985).] In a blodegradatlon screening study using
activated sewage as seed. 3,3'-d1chlorobenz1d1ne at an Initial concentration
of 3 mg/l underwent 9-99* degradation 1n 28 days when yeast extract was
present at concentrations of 10-400 mg/l. Extent of degredatlon was
dependent on the concentration of yeast extract and no degradation was
observed In this absence of this additional nutrient (Brown and Laboureur,
1983). As a result, 3,3'-d1chlorobenz1d1ne was considered to be "Inherently
biodegradable" rather than "readily biodegradable". Brown and Laboureur
(1983) suggested that the yeast extract may have provided growth factors
necessary for the breakdown of this amlne or It might have been acting as a
readily degradable food source, building up a large concentration of active
bacteria, which were then able to break down 3,3'-d1chlorobenz1d1ne. In
another blodegradatlon screening study, 2.7X degradation (based on SCO-
evolved) of 0.05 mg/l 3,3'-d1chlorobenz1d1ne was observed after 5 days
Incubation In activated sludge (Freltag et al., 1985).
0087d -6- 03/31/88
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2.2.5. Volatilization. Henry's Law constant for 3,3'-d1chlorobenz1d1ne
was estimated to be ~5xlO~8 atm-m3/mo1 at 25°C based on a water solubil-
ity of 3 mg/l at 25°C and an estimated vapor pressure of 4.2xlO~7 mm Hg
at 25°C. This value for Henry's Law constant Indicates that this compound
1s essentially nonvolatile and that volatilization would not be a signifi-
cant fate process In water (Lyman et al.t 1982).
2.2.6. B1oaccumulat1on. 3,3'-D1chlorobenz1d1ne was found to be rapidly
and significantly bloaccumulated by fish and algae. Appleton and Slkka
(1980) exposed blueglll sunflsh, Lepomls macrochlrum. to water containing 5
and 100 wg/a radlolabelled 3,3'-d1chlorobenz1d1ne until reaching
equilibrium (4-7 days). Based on total 14C residues found at equilibrium,
the apparent BCF of 3,3'-d1chlorobenz1d1ne was determined to be 495-507 In
whole fish, 114-170 In the edible portion and 814-856 1n head and viscera.
Appleton and Slkka (1980) suggested the possibility that either
enterohepatlc circulation of 3,3'-d1chlorobenz1d1ne and metabolites or
covalent binding to Upoproteln may account for some of the residual
radioactivity detected. In another study, a BCF of 610 was found In Golden
Ide, Leudscus Idus melanotus. exposed to 50 yg/l radlolabelled
3,3'-d1chlorobenz1d1ne for 3 days; a BCF of 940 was found In a green algae.
Chlorella fusca. exposed to 50 »ig/l radlolabelled 3,3'-d1chlorobenz1d1ne
for 1 day (Freltag et al., 1985).
2.2.7. Adsorption. The relative distribution ratio of 3,3'-d1chloroT
benzldlne between natural aquatic sediments and water at pH 5-7 was found to
range between 26.7 and 128 (Appleton et al., 1978). Adsorption to sediments
was Inhibited by 30-50X under alkaline conditions (pH 9). Equilibration of
3,3'-d1chlorobenz1d1ne between water and sediment was generally achieved
within 24 hours and desorptlon from sediments was very low. Based on the
behavior of other aromatic amines 1t was speculated that Initial adsorption
0087d -7- 03/31/88
-------
of 3,3'-d1chlorobenz1d1ne to sediments was probably the result of physical
adsorption processes, which was then followed by covalently binding with the
sediment. Irreversible binding with sediment would explain the Increased
resistance of 3,3'-d1chlorobenz1d1ne to desorptlon and extraction observed
over time (Slkka et al., 1978).
2.3. SOIL
2.3.1. Hydrolysis. 3,3'-D1chlorobenz1d1ne contains no hydrolyzable
functional groups (Callahan et al., 1979); therefore, this compound 1s not
expected to undergo chemical hydrolysis In soil.
2.3.2. Oxidation. UnsubstUuted benzldlne 1s rapidly oxidized by Fe* ,
Al* , Cu* and a few other naturally occurring cations, which can be
found 1n soil as part of humlc add complexes and mlcrocrystalUne clays
(Callahan et al., 1979; Demlrjlan et al., 1987). Whether or not 3,3'-d1-
chlorobenzldlne would be oxidized rapidly In a similar manner 1s not
certain; however, the possibility exists that 3,3'-d1chlorobenz1d1ne may be
oxidized by catlonlc components of soil (see Section 2.2.2.).
2.3.3. Photolysis. Given that 3,3'-d1chlorobenz1d1ne photogrades rapidly
In water and when adsorbed onto silica gel (see Sections 2.1.3. and 2.2.3.)
1t Is also expected to photodegrade rapidly on soil surfaces.
2.3.4. M1crob1al Degradation. A Brookston clay loam soil was mixed with
l4C-3,3'-d1chlorobenz1d1ne (total concentrations of 4 or 40 ppm were
achieved In 2 separate batches) and Incubated 1n the dark under aerobic
conditions for 32 weeks or under anaerobic conditions for 1 year.
Cumulative 14CO_ production was ~2X after 32 weeks Incubation In aerobic
soil containing 4 and 40 ppm 3,3'-d1chlorobenz1d1ne. No radioactive
14CH. or 14COp was detected 1n the headspace gas above the anaerobic
soil after 1 year of Incubation (Boyd et al., 1984).
0087d -8- 03/31/88
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2.3.5. Adsorption. The K for 3,3'-d1chlorobenz1d1ne was experiment-
ally determined to be 16,300 1n Brookston clay loam and 33,700 In' Rubicon
sand (Boyd et al., 1984). These K values Indicate that strong physical
adsorption of 3,3'-d1chlorobenz1d1ne to soil takes place. 3,3'-01chloro-
benzldlne has also been shown to bind strongly to soil. When radlolabelled
3,3'-d1chlorobenz1d1ne (at 4 and 40 ppm concentration) were added to
Brookston clay loam, >50X of the 14C was nonextractable after the first
several weeks of Incubation. In general, at least 50% of the aromatic amines
became bound to soil 1 day to 1 week. Aromatic amines such as 3,3'-d1-
chlorobenzldlne are believed to form covalent linkages with humlc substances
1n soil, thus Immobilizing them In soil. Two different mechanisms have been
proposed for the chemical binding of amines to soil (Boyd et al., 1984).
Rapid reversible binding of primary amines with humate carbonyls Is believed
to result 1n Imlne formation. Subsequent slow Irreversible reaction thought
to represent !,4-add1t1on to qulnone rings 1s believed to occur. This
addition would result In an amlno-substltuted qulnone. The amlne group can
be further converted to heterocycllc nitrogen, which Is present In the
humate structure. These reactions may proceed enzymatlcally and chemically
(Boyd et al., 1984).
2.3.6. Volatilization. Volatilization of 3,3'-d1chlorobenz1d1ne from
soil surfaces 1s expected to be negligible since this compound binds/adsorbs
strongly to soil and has a relatively low Henry's Law constant (see Sections
2.2.5. and 2.3.3.). At the end of 1 year Incubation of labeled
3,3'-d1chlorobenz1d1ne and soil, essentially all the original 14C still
remained In the soil, demonstrating that volatile losses of
3,3'-d1chlorobenz1d1ne or Us metabolites had not occurred 1n this system
(Boyd et al., 1984).
0087d -9- 03/31/88
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2.4. SUMMARY
In the atmosphere, 3,3'-d1chlorobenz1d1ne Is expected to exist primarily
In the participate form. 3,3'-01chlorobenz1d1ne, In both vapor and partlcu-
late form, Is expected to undergo rapid photolysis In the atmosphere.
Reaction of 3,3'-d1chlorobenz1d1ne vapor with photochemically generated
hydroxyl radicals (t,._=10 hours) (Atkinson, 1987) may be a minor removal
mechanism. In water, this compound would undergo rapid photodegradatlon
(t,/2=90 sec) 1n the surface layers of water (Banerjee et al., 1978).
This compound photodegrades to monochlorobenzldlne, benzldlne and a number
of brightly colored water-Insoluble compounds (Banerjee et al., 1978).
Beyond the reach of light penetration, this compound would rapidly adsorb
to sediment and partlculate matter where 1t Is tightly bound. Adsorption Is
expected to proceed Initially by a rapid, but reversible, physical adsorp-
tion process followed by much slower Irreversible covalent bonding (Appleton
et al., 1978; Slkka et al., 1978). It has also been speculated that
3,3'-d1chlorobenz1d1ne may be oxidized by naturally occurring cations such
as, Fe found 1n sediments (Callahan et al., 1979). Rapid uptake and
bloaccumulatlon In aquatic organisms 1s also expected to occur (Appleton and
Slkka, 1980; Freltag et al.. 1985). Volatilization, mlcroblal degradation
and chemical hydrolysis are not expected to be Important fate processes In
water. In soil, 3,3'-d1chlorobenz1d1ne Is expected to adsorb tightly to
soil, and over time, Irreversibly bind with humates In the soil (Boyd et
al., 1984). It has also been speculated that oxidation by reaction with
catlonlc constituents of soil such as, Fe* and Cu* may also occur
(Callahan et al., 1979; Dem1rj1an et al., 1987). If exposed to sunlight on
soil surfaces, 3,3'-d1chlorobenz1c!1ne 1s expected to photodegrade rapidly.
Volatilization and mlcroblal degradation are not expected to be significant
fate processes.
0087d -10- 03/31/88
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3. EXPOSURE
Exposure to 3,3'-d1chlorobenz1d1ne 1s most likely to occur In occupa-
tional settings, particularly where this compound 1s manufactured or where
3,3'-d1chlorobenz1d1ne-based dyes are manufactured or used (HSDB, 1987). In
1973, 18 U.S. companies were using 3,3'-d1chlorobenz1d1ne and 166-250
workers were potentially exposed to this compound (IARC, 1982). The general
public may be exposed to 3,3'-d1chlorobenz1d1ne during use of paints, pig-
ments or enamels derived from this compound. 3,3'-D1chlorobenz1d1ne occurs
at a level of -20 ppm 1n most pigments (Lapp et al., 1981). The most prob-
able routes of exposure are Inhalation of dusts or mists and dermal contact
(HSDB, 1987).
3.1. WATER
3,3'-D1chlorobenz1d1ne has been detected 1n samples of surface water,
sediment, fish and Industrial effluent collected throughout the United
States. Monitoring data are provided 1n Table 3-1.
3.2. FOOD
Pertinent monitoring data regarding the presence of 3,3'-d1chloro-
benzldlne 1n food were not located In the available literature cited In
Appendix A.
3.3. INHALATION
Limited data were available concerning the detection of 3,3'-d1chloro-
benzldlne In air. Results of a 1970 Japanese study on worker exposure to
3,3'-d1chlorobenz1d1ne 1n a pigment manufacturing plant revealed that the
concentration of 3,3'-d1chlorobenz1d1ne In the air reached a level of 25
ug/m3 (2 ppb) within 10 minutes of charging reaction vessels and dropped
to 2 wg/ma (0.2 ppb) within 20 minutes (IARC, 1982).
0087d -11- 03/31/88
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TABLE 3-1
0
o
oo
~j
Q.
Sample Type
Total water
Sediment3
Sediment3
- Fish tissue
ro
i
Surface water
Surface water
i
Sediment/soil/
Monitoring Data for 3t3'-D1chlorobenz1dtne In Various Media
Location
United States
United States
United States
United States
Buffalo River.
Cayuhoga River,
St. Joseph's River
Sumlda R1verc
(Japan)
Love Canal
Number of
Samples
7334
952
68
650
NR
NR
NR
Concentration
46 yg/l (mean)
3222 Mg/kg dry wt.
basis (mean)
0.026 rug/kg wet wt.
basis (mean)
5.8 mg/kg wet wt.
basis (mean)
not detected**
qualitatively
Identified
qualitatively
Reference
U.S. EPA. 1987b
U.S. EPA, 1987b
U.S. EPA. 1987b
U.S. EPA, 1987b
Great Lakes Water
Quality Board. 1983
IARC, 1982
Hauser and
water
Purge wells and
seepage water
o
en
CO
oo
(Niagara Falls,
NY); 1980
near a waste
disposal lagoon
receiving waste
from the manu-
facture of 3,3'-
dlchlorobenzldlne
NR
Identified
0.13-0.27 mgA
Bromberg. 1982
IARC. 1982
-------
o
CO
TABLE 3-1 (cont.)
Sample Type
Location
Number of
Samples
Concentration
Reference
Treated effluent
from coal mining
Treated effluent
from nonferrous
metal manufacturing
United States
United States
52 (2% pos.) 3 Pg/l
18
0.2 pg/t (mean)
U.S. EPA, 1981
U.S. EPA, 1981
Treated effluent"
Urban runoff6
United States
15 United
States cities
NR
86
not detected0
not detected
U.S. EPA, 1981
Cole et al., 1984
aSed1ment samples were analyzed on either a wet or dry weight basis.
^Detection limit not reported
cTh1s River receives wastewater from several dye and pigment factories. '
, dTreated effluent from leather tanning and finishing, aluminum forming, battery manufacturing, coll
coating, foundarles, porcelain enameling, gum and wood chemicals, pharmaceutical manufacturing, organic
chemicals manufacturing/plastics, pulp and paperboard mills, rubber processing, steam electric power
plants, timber products processing.
eU.S. EPA Nationwide Urban Runoff Program Findings as of July 1982.
NR = Not reported
o
IS)
CD
CO
-------
3.4. DERMAL
Pertinent monitoring data regarding dermal exposure to 3,3'-d1chloro-
benzldlne were not located 1n the available literature cited In Appendix A.
3.5. SUMMARY
Exposure to 3,3'-d1chlorobenz1d1ne 1s most likely to occur 1n occupa-
tional settings, particularly where this compound 1s manufactured or where
3,3'-d1chlorobenz1d1ne-based dyes are manufactured or used (HSDB, 1987).
The general public may be exposed to low levels of 3,3'-d1chlorobenz1d1ne
during use of paints, pigments or enamels derived from this compound. The
most probable routes of exposure are Inhalation of dusts or mists containing
this compound and dermal contact (HSDB, 1987). 3,3'-D1chlorobenz1d1ne has
been detected In samples of surface water, sediment, fish and Industrial
effluent (U.S. EPA, 1981, 1987b; IARC, 1982; Hauser and Bromberg, 1982).
This compound has not been detected 1n urban runoff samples collected as
part of the U.S. EPA Nationwide Urban Runoff Program (Cole et al.t 1984).
The U.S. EPA STORET Data Base Indicates that the mean concentrations of
3,3'-d1chlorobenz1d1ne In total water, sediment (on either wet or dry weight
basis) and fish tissue samples collected throughout the United States are 46
vig/l, 3222 yg/kg dry wt. of sediment, 0.026 mg/kg wet wt. of sediment,
and 5.8 mg/kg wet wt. of fish tissue, respectively (U.S. EPA, 1987b).
0087d -14- 05/10/88
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4. AQUATIC TOXICITY
There are few data regarding the aquatic toxldty of 3,3'-d1chlorobenz1-
dlne. A study by Appleton et al. (1978) Indicated that 1n bluegllls,
Lepomls macrochlrus. toxic levels of 3,3'-d1chlorobenz1d1ne accumulated
before a 3,3'-d1chlorobenz1d1ne equilibrium was reached between water and
fish. There were many mortalities when the whole body residues of
l4C-3,3'-d1chlorobenz1d1ne exceeded 150 ppm.
0087d -15- 03/31/88
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5. PHARMACOKINETICS
5.1. ABSORPTION
The absorption of 3,3'-d1chlorobenz1d1ne following administration of the
compound by relevant routes (I.e., oral and Inhalation) has not been studied
extensively. Hsu and S1kka (1982) reported that 3,3'-d1chlorobenz1d1ne Is
absorbed readily following oral administration of the compound to rats. The
kinetics of the appearance of radio-activity 1n the plasma monitored 1n rats
following a single oral dose of radlolabeled 3,3'-d1chlorobenz1d1ne (40
mg/kg) In DMSO. Total plasma radioactivity reached a peak (9.4 yg
3,3'-d1ch1orobenz1d1ne equivalents/ml) 8 hours after dosing and then
declined In a blphaslc manner. The appearance and disappearance of total
radioactivity In the plasma could be described by a first-order absorption
process with a half-life of 1.59 hours for the appearance and 1.68 and 33
hours, respectively, for the two elimination process. The kinetic pattern
of unchanged 3,3'-d1chlorobenz1d1ne 1n plasma was similar to that for total
radioactivity but reached a peak (1.25 yg 3,3'-d1chlorobenz1d1ne/m8.) 4
hours after dosing and after which time 1t declined In a blphaslc manner.
The absorption of unchanged 3,3'-d1chlorobenz1d1ne was also first-order with
a reported half-life of 2.12 hours for the appearance and 5.58 and 13.59
hours for the disappearance processes.
Data regarding the excretion of 3,3'-d1ch1orobenz1d1ne and metabolites
1n the urine, bile and feces of rats given an oral dose of 40 mg 3,3'-d1-
chlorobenzldlne/kg Indicated that >90X of the administered dose was absorbed
(Hsu and Slkka, 1982).
Pertinent data regarding the absorption of 3,3'-d1chlorobenz1d1ne
following Inhalation exposure 1n humans or animals were not located In the
available literature.
0087d -16- 04/01/88
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There appears to be significant absorption of 3,3'-d1ch1orobenz1d1ne
following dermal exposure to the compound In both humans and animals. An
early study (Melgs et al., 1954) of occupational exposure to 3,3'-dlchloro-
benzldlne Indicated that the skin was the principal route of entry of this
compound 1n exposed workers. Susklnd (1983) also reported that absorption
of 3,3'-d1chlorobenz1d1ne through the skin 1s the major route of entry Into
the body. The dermal absorption of 3,3'-dlchlorobenzldlne 1n rats was
studied by Shah and Guthrle (1983), who applied 0.2 ma of "C-S.S'-dl-
chlorobenzldlne In acetone (total dose 1 mg/kg) to the shaved backs of
Fisher 344 rats. The amount of 3,3'-dlchlorobenzldlne absorbed from the
application site after 24 hours was -50% of the applied dose (half-life of
disappearance of radioactivity from the application site was estimated to be
24.1 hours).
5.2. DISTRIBUTION
The distribution of radioactivity In rats given either single or
multiple oral doses of l4C-rad1olabeled 3,3'-dlchlorobenzldlne was studied
by Hsu and S1kka (1982). Rats were given a single oral dose of either 6.4
or 40 mg 3,3'-d1chlorobenz1d1ne/kg, and various tissues were analyzed for
radioactivity 24 or 96 hours after dosing. At both dose levels and at both
sacrifice times, the liver, lung and kidney were the principal sites of
distribution of 3,3'-d1chlorobenz1d1ne-der1ved radioactivity. At the 40 mg
3,3'-d1chlorobenz1d1ne/kg dose level, the concentration of radioactivity 1n
the principal organs of accumulation reached maximum levels 12-16 hours
following dosing; maximum levels for the various organs were liver, 53.4
wg 3,3'-d1chlorobenz1d1ne equlvalents/g tissue; kidney, 36.9 yg 3,3'-d1-
chlorobenzldlne equlvalents/g tissue; and lung, 16.1 vg 3,3'-dlchloro-
benzldlne equlvalents/g tissue. The measured half-lives for the appearance
of radioactivity 1n these tissues were 3.32, 2.5 and 5.41 hours, respec-
0087d -17- 03/31/88
-------
tlvely, for the liver, lung and kidney. In the same study, multiple oral
dosing of rats with 3,3'-d1chlorobenz1d1ne (6 mg/kg/day for 6 days) led to
tissue distributions of radioactivity similar to those observed after single
doses (I.e., the highest concentrations of radioactivity were found In the
liver and kidney). The tissue concentrations of radioactivity were, how-
ever, 3-4 times higher In rats given multiple doses. Multiple oral dosing
with 3,3'-d1chlorobenz1d1ne did not result In substantial retention of
radioactivity, and Hsu and S1kka (1982) concluded that the compound had a
fairly low tendency to accumulate In the body.
The distribution of radioactivity In the tissues of rats following
dermal application of a solution of 3,3'-d1chlorobenz1d1ne in acetone (total
3,3'-d1chlorobenz1d1ne dose of 1 mg/kg) was studied by Shah and Guthrle
(1983). For most tissues the amount of radioactivity recovered 24 hours
after 3,3'-d1chlorobenz1d1ne application-was <0.1X of the dose. The liver^
however, showed the highest level of radioactivity, ~4% of the applied dose.
5.3. METABOLISM
The limited data available regarding the metabolism of 3,3'-d1chloro-
benzldlne do not provide sufficient Information to suggest a metabolic
scheme for the compound. Troll (n.d.) was unable to detect ortho-hydroxy
metabolites of 3,3'-d1chlorobenz1d1ne In the urine of humans and dogs after
oral dosing and Shrlner et al. (1978) suggested that the chlorine constitu-
ents of 3,3'-d1chlorobenz1d1ne probably block ring hydroxylatlon reactions
for both electronic and sterlc reasons. Hsu and S1kka (1982), however, have
provided evidence that 3,3'-d1chlorobenz1d1ne Is metabolized extensively In
rats. These Investigators found at least five metabolites of
3,3'-d1chlorobenz1d1ne 1n ether extracts of urine and bile samples from rats
24 hours after treatment with an oral dose of radlolabeled 3,3'-d1chloro-
benzldlne (40 mg/kg). Although the metabolites were not Identified, It was
0087d -18- 03/31/88
-------
determined that only 9 and 16% of the total radioactivity excreted In the
urine and bile, respectively, could be accounted for by the parent
compound. Therefore 3,3'-d1chlorobenz1d1ne has been demonstrated to be
metabolized extensively following absorption 1n this study.
Kellner et al. (1973) reported the presence of a compound with chromato-
graphlc properties similar to mono-N-acetyl 3,3'-d1chlorobenz1d1ne 1n the
urine of monkeys dosed Intravenously with 14C-labeled 3,3'-d1chlorobenz1-
dlne (0.2 mg/kg). Aksamltnala (1959) reported the appearance of four
transformation products In the urine of rats given repeated oral doses of
3,3'-d1chlorobenz1d1ne over a period of 7.5-8.5 months. The products of
3,3'-d1chlorobenz1d1ne metabolism. Identified only by paper chromatography,
were tentatively Identified as benzldlne and some possible glucuronlde
conjugates.
Several studies are available regarding the In vitro metabolism of
3,3'-d1chlorobenz1d1ne. Cytochrome c was reported to be Incapable of
oxidizing 3,3'-d1chlorobenz1d1ne, whereas benzldlne and several other
derivatives were oxidized (H1ra1 and Yasuhlra, 1972). Studies of the
binding of radlolabeled 3t3'-d1chlorobenz1d1ne to calf thymus ONA Indicated
that 3,3'-d1chlorobenz1d1ne reacted with ONA both In the absence and
presence of rat liver S-9 fraction. Binding 1n the presence of S-9 was
considerably higher than that observed 1n Us absence (Bratcher and S1kka,
1982).
5.4. EXCRETION
The elimination of 3,3'-d1chlorobenz1d1ne and 3,3'-d1chlorobenz1d1ne-
derlved radioactivity from the plasma and principal organs of distribution
(liver, lung and kidney) was characterized In rats treated with single oral
doses of 14C-labeled 3,3'-d1chlorobenz1d1ne (Hsu and S1kka, 1982). The
disappearance of both the parent compound and total radioactivity from the
0087d -19- 03/31/88
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plasma of 3,3'-d1chlorobenz1d1ne-treated rats was blphaslc. The elimi-
nation half-lives for total radioactivity from the plasma were 1.68 and 33
hours for the fast and slow phases, respectively. The corresponding values
for the fast and slow phases of elimination of the parent compound from the
plasma were 5.58 and 13.59 hours, respectively. The disappearance of radio-
activity from the major tissues of distribution was also blphaslc 1n nature.
Half-lives for the slow and fast phases were liver, 5.78 and 77 hours; lung,
3.85 and 43.3 hours; and kidney, 7.14 and 138.6 hours.
The bile appears to be a significant route of excretion of 3,3'-d1-
chlorobenzldlne and metabolites. Preliminary experiments with rats
Indicated that >90% of the administered radioactivity was excreted via the
urine and feces 72 hours after receiving single oral doses of radlolabeled
3,3'-d1chlorobenz1d1ne (40 mg/kg) (Hsu and Slkka, ,1982). Of this
3,3'-d1chlorobenz1d1ne-der1ved radioactivity, 64.9% was found 1n the feces
and 27.7% of the administered radioactivity was found In the urine. To
determine the source of the radioactivity appearing 1n the feces, further
experiments were performed using bile duct-cannulated rats given oral doses
of 14C-labeled 3,3'-d1chlorobenz1d1ne. Cumulative excretion of radio-
activity was monitored 1n the urine, feces and bile for up to 72 hours after
dosing. Excretion of radioactivity In the urine, bile and feces again
approached 90% of the administered radioactivity; however, -65% of the
administered dose was found In the bile within 24 hours of dosing compared
to 48% 1n the Intact rats. Excretion of radioactivity In the urine and
feces decreased to <10% of the administered dose for each route compared to
27.7% and 64.9%, respectively, 1n Intact rats. Hsu and Slkka (1982)
concluded that biliary excretion Is a significant route of elimination for
3.3'-d1chlorobenz1d1ne and Us metabolites.
0087d -20- 04/01/88
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Support for the Importance of hepatoblHary excretion 1n the elimination
of 3,3'-d1chlorobenz1d1ne Is provided by a study by Kellner et al. (1973).
Following Intravenous administration of a radlolabeled dose of 3,3'-d1-
chlorobenzldlne to rhesus monkeys, most of the administered dose was
recovered from the bile, Intestine and liver within 14 hours of treatment.
Troll (n.d.) reported that In humans 3,3'-d1chlorobenz1d1ne Is excreted
largely 1s the feces. Several Investigators have also reported detectable
quantities of 3,3'-d1chlorobenz1d1ne 1n the urine of 3,3'-d1chloro-
benzldlne processing and manufacturing workers (Melgs et al., 1954; Slngal
and Lee. 1985; London and Bolano, 1986).
5.5. SUMMARY
The absorption of 3,3'-d1chlorobenz1d1ne following administration of the
compound by relevant routes (Inhalation or oral exposure) has not been
studied extensively. Hsu and S1kka (1982) reported that 3,3'-d1chlorobenz1-
dlrie 1s rapidly absorbed and extensively distributed following oral
administration of the compound to rats. Helgs et al. (1954) and Susklnd
(1983) reported that the skin 1s the most significant route of entry of
3,3'-d1chlorobenz1d1ne Into the body In cases of occupational exposure. The
half-life of d1s- appearance of a topically applied 3,3'-d1chlorobenz1d1ne
dose from the shaved backs of rats was determined to be 24.1 hours (Shah and
Guthrle, 1983).
Following a single oral dose of radlolabeled 3,3'-d1chlorobenz1d1ne to
rats, the principal organs In which radioactivity was found were the liver*
kidneys and lungs (Hsu and S1kka, 1982). Multiple oral 3,3'-dlchlorobenzl-
dlne dosing led to tissue levels of radioactivity 3-4 times higher than the
levels observed following a single oral dose, but a multiple dosing schedule
did not result In substantial retention of radioactivity (Hsu and Slkka,
1982).
0087d -21- 03/31/88
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The extent of metabolism and the pathways of 3,3'-d1chlorobenz1d1ne
metabolism are not clear. Ring hydroxylatlon products of 3,3'-d1chloro-
benzldlne were not found 1n the urine of humans and dogs given an oral
3,3'-d1chlorobenz1d1ne dose (Troll, n.d.), and Shrlner et al. . (1978)
suggested that chlorlnatlon of benzldlne blocks ring hydroxylatlon reactions
of 3,3'-d1chlorobenz1d1ne for both electronic and sterlc reasons. Hsu and
Slkka (1982), however, provided evidence that 3,3'-d1chlorobenz1d1ne Is
metabolized extensively In rats.
Possible metabolites of 3,3'-d1chlorobenz1d1ne, tentatively Identified
by chromotographtc procedures, Include mono-N-acetyl 3,3'-d1chlorobenz1-
dlne 1n the urine of monkeys (Kellner et al., 1973) and benzldlne and some
possible glucuronlde conjugates 1n the urine of rats (Aksamltnala, 1959).
Elimination of both radlolabeled 3,3'-d1chlorobenz1d1ne and total radio-
activity from the plasma of orally dosed rats was blphaslc (Hsu and Slkka,
1982). Similarly, blphaslc mode of elimination of total radioactivity was
observed 1n the principal organs of distribution (liver, lung and kidney)
(Hsu and Slkka, 1982).
The bile appears to be a significant route of excretion for both
3,3'-d1chlorobenz1d1ne and Us metabolites. Experiments using rats
Indicated that -90% of the administered radioactivity Is excreted In the
urine and feces following an oral dose of 3,3'-d1chlorobenz1d1ne (Hsu and
Slkka, 1982). Approximately 65% of the administered radioactivity was
excreted 1n the feces and the major source of the radioactivity found 1n the
feces originated from the bile. Hepatoblllary excretion of
3,3'-d1chlorobenz1d1ne and Us metabolites also occurred In rhesus monkeys
(Kellner et al., 1973).
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The fecal route appears to be the most significant route of elimination
of 3,3'-d1ch1orobenz1d1ne and metabolites In humans (Troll, n.d.). 3,3'-d1-
chlorobenzldlne has also been reported to be present 1n the urine of occupa-
tional ly-exposed Individuals (Helgs et al., 1954; London and Bolano, 1986;
Slngal and Lee, 1985).
0087d -23- 03/31/88
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6. EFFECTS
6.1. SYSTEMIC TOXICITY
6.1.1. Inhalation Exposures. Pertinent data regarding the systemic
toxldty of 3,3'-d1chlorobenz1d1ne following subchronlc or chronic Inhala-
tion exposure In either animals or humans were not located 1n the available
literature cited 1n Appendix A.
6.1.2. Oral Exposures.
6.1.2.1. SUBCHRONIC -- Pertinent data regarding the systemic toxldty
of 3,3'-d1chlorobenz1d1ne following subchronlc oral exposure 1n either
animals or humans were not located 1n the available literature cited In
Appendix A.
6.1.2.2. CHRONIC — Six female beagle dogs were given 3,3'-d1chloro-
benzldlne (1QQ mg/day, -100% pure) by capsule 3 times/week for 6 weeks, then
5 times/week continuously for an additional 7 years (total duration = 7.1
years) (Stula et a!., 1978). Six untreated female beagle dogs served as
controls and were sacrificed after 8.3-9.0 years. Urine and blood samples
were taken once before the test began and then approximately every 6 months
during the remainder of the test. At sacrifice, a complete necropsy and
hlstologlcal examination were performed on all dogs. All six 3,3'-d1chloro-
benzldlne-treated dogs had elevated SGPT activities during the first 3 years
of treatment. SGPT activities remained elevated 1n two of four dogs that
survived the full treatment period (7.1 years).
6.1.3. Other Relevant Information. The oral LD 5Q of In albino rats
(sex and strain not specified) was reported to be -7 g/kg bw (ACGIH, 1986)
for 3,3'-d1chlorobenz1d1ne and 3.82 g/kg bw for dlhydrochlorlde salt of
3,3'-d1chlorobenz1d1ne.
0087d -24- 04/01/88
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Human Mbroblast cell cultures were found to be more sensitive than
hamster flbroblast cell cultures to the cytotoxlc effects due to
3,3'-d1chlorobenz1d1ne exposure (Casto, 1983). The LC-. following 18
hours of exposure to 3,3'-d1chlorobenz1d1ne was determined to be 250
yg/mt for hamster cells and 50 yg/ms. for human cells.
6.2. -CARCINOGENICITY
6.2.1. Inhalation. Pertinent data regarding the carclnogenldty of
3,3'-d1chlorobenz1d1ne following Inhalation exposure In either animals or
humans were not located 1n the available literature cited In Appendix A.
6.2.2. Oral. One of the first demonstrations of the carclnogenldty of
3,3'-d1chlorobenz1d1ne following oral administration 1n rats was a study by
PUss (1959). Fifteen female and 35 male outbred Rappolovo rats were fed
3,3'-d1chlorobenz1d1ne In a paste (45.3% 3,3'-d1chlorobenz1d1ne, 50% water
and 4.7% unspecified Impurities) 1n an amount that provided a dose of 10-20
mg/day. The paste was administered 6 days/week for 12 months (total dose of
4.5 g 3,3'-d1chlorobenz1d1ne/rat) and the rats were observed for life.
Control animals (130 rats) were Injected with either octadecylamlne or
methylstearylamlne and observed for 23 months. The numbers of rats that
survived were: 34 at 6 months, 29 at the time of appearance of the first
tumor (11 months) and 27 at 12 months. Twenty-three out of 50
3,3'-d1chlorobenz1d1ne-treated rats developed tumors. There were seven
tumors of the Zymbal gland, three skin tumors, seven mammary gland tumors,
two adenocarclnomas of the 11eum, three bladder tumors, three tumors of the
haematopoietic system, two tumors of the connective tissue, two salivary
gland tumors, one liver tumor and one thyroid tumor. Incidences, expressed
as the number of rats with a particular tumor type per number of rats
examined, were not provided. No tumors were found In the control group.
However, the lack of adequate controls was noted In IARC (1982).
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Fifty female and fifty male ChR-CD rats were fed 3,3'-d1chlorobenz1d1ne
1n the diet (1000 ppm 3,3'-d1chlorobenz1d1ne, purity and Impurities unspeci-
fied) for an average period of 349 days (range of 143-488 days) for females
and 353 days (range of 118-486 days) for males (Stula et al., 1975).
Control rats (50 male and 50 female) were fed a standard diet and were main-
tained under observation for up to -2 years. Six 3,3'-d1chlorobenz1d1ne-
treated rats/sex were sacrificed at 12 months and were not Included 1n the
tumor analysis. Of the remaining forty-four 3,3'-d1chlorobenz1d1ne-treated
rats of each sex, statistically significant (p<0.05) Increases 1n tumor
Incidences over those observed 1n controls were reported (Table 6-1).
3,3'-D1chlorobenz1d1ne-treated male rats had significantly Increased
Incidences of granulocytlc leukaemias, mammary adenocardnomas and Zymbal
gland carcinomas. 3,3'-D1chlorobenz1d1ne-treated female rats had a
significantly Increased Incidence of mammary adenocardnomas.
The ability of 3,3'-d1chlorobenz1d1ne dlhydrochlorlde to produce mammary
tumors 1n young female Sprague-Dawley rats (40 days old) was Investigated by
Grlswold et al. (1968). A group of 20 rats were given 10 doses of
3,3'-d1chlorobenz1d1ne dlhydrochlorlde by gastric Intubation every 3 days,
which was a total administered 3,3'-d1chlorobenz1d1ne dose of 300 mg/rat
over a 30-day period. Rats were observed for 9 months following treatment.
Fourteen 3,3'-d1chlorobenz1d1ne-treated rats survived to the end of the
9-month observation period. Negative controls were administered only with
the vehicle (sesame oil) and positive controls received a single dose of 18
mg DMBA. At necropsy and hlstologlcal examination, none of the
3,3'-d1chlorobenz1d1ne-treated rats had mammary tumors, while the Incidence
of mammary tumors was 100% In the DMBA controls and 3% In the negative
control group.
0087d -26- ~ 05/10/88
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o
GO
TABLE 6-1
incidence of Tumors in Female Beagle Dogs Given 3,3-Dlchlorobenzidine (-100X pure) Orally by Capsule3
00
Dose
100 mg/day,
for 6 weeks,
5 times /week
3 times/week
followed by
for 7 years
Duration
of Study
(years)
<7.1
Target Organ
liver
urinary bladder
Tumor Type
carcinoma
papillary transi-
tional cell carcinoma
Tumor Incidence**
(p value)0
4/5
(p<0.025)
5/5
(p<0.025)
0 (untreated controls)
Strength of study:
' Weakness of the study:
Overall adequacy:
8.3-9.0 liver or NA 0/6
urinary bladder
QUALITY OF EVIDENCE
Compound was administered by a relevant route of exposure for a sufficient
duration for tumor development.
Only six dogs were started on the study and only five survived to be at risk for
late appearing tumors; only one dose level was administered to only one sex.
Adequate
aSource: Stula et al.. 1978
DTumor Incidence expressed as number of animals with tumors/number of animals necropsied
cp value is for Fisher Exact test (one tall)
NA = Not applicable
-------
Six female beagle dogs were given a dally oral dose of 100 mg 3,3'-d1-
chlorobenzldlne (-100% pure) by administration of a capsule 3 times/week for
6 weeks, then 5 times/week for up to an additional 7 years (total duration
equal to 7.1 years) (Stula et a!., 1978). Six untreated beagle dogs served
as controls. A complete necropsy and histologlcal examination were per-
formed on all dogs at the end of the test period. One 3,3'-d1chlorobenz1-
dlne-treated dog, sacrificed \jn extremis after 3.5 years, had no tumors.
Another 3i3'-d1chlorobenz1d1ne-treated dog, sacrificed in extremis after 6.6
years, developed an undlfferentlated carcinoma of the liver and a papillary
transitional cell carcinoma of the urinary bladder. Of the four 3,3'-d1-
chlorobenzldlne-treated dogs that survived the full test period of 7.1
years, all had papillary transitional cell carcinomas of the urinary bladder
and three had hepatocellular carcinomas. None of the six control dogs had
liver or urinary bladder tumors (Table 6-2).
Syrian golden hamsters (30 male and 30 female) were fed 0.1% (w/w)
3,3'-d1chlorobenz1d1ne 1n the diet (40% dlhydrochlorlde. 60% free base) ad
libitum for their lifetime (Saff1ott1 et al., 1967). A similar group of
controls received an untreated diet. The average food Intake was 60
g/hamster/week and this led to a calculated 3,3'-d1ch1orobenz1d1ne dose of
60 mg 3.3'-d1chlorobenz1d1ne/hamster/week. 3,3'-D1chlorobenz1d1ne adminis-
tration failed to produce any significant carcinogenic effect or bladder
pathology 1n the treated hamsters. In a follow-up study by the same group
of Investigators (Sellakumar et al., 1969) a similar number of hamsters was
fed a diet containing 0.3% (w/w) 3,3'-d1chlorobenz1d1ne. At this dietary
level, 3,3'-d1chlorobenz1d1ne was reported to have Induced four transitional
cell bladder carcinomas, some liver-cell and cholanglomatous tumors and
diffuse chronic 1ntrahepat1c obstructing cholangltls. This study
0087d -28- 03/31/88
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TABLE 6-2
Incidence of Tumors 1n Chr-CD Rats Treated with 3,3-D1chlorobenz1d1ne
In the Diet3
Sex
H
F
H
Duration of
Dose Treatment^
(ppm) (days)
1000 353
1000 349
0 564
Target
Organ
blood
mammary
skin
mammary
blood
mammary
skin
Tumor Type
granulocytlc
leukemia
adenocardnoma
Zymbal gland
carcinoma
adenocardnoma
granulocytlc
leukemia
adenocardnoma
Zymbal gland
Tumor Incidence0
(p value)d
9/44 (p<0.05)
7/44 (p<0.05)
8/44 (p<0.05)
26/44 (p<0.05)
2/44
0/44
0/44
628
carcinoma
mammary adenocardnoma
3/44
Strength of study:
Weakness of study:
Overall adequacy:
QUALITY OF EVIDENCE
Compound was administered by a relevant route of
exposure to a sufficient number of both sexes of one
species for a sufficient duration.
Only one dose level was used.
Adequate
aSource: Stula et al.. 1975
bDurat1on of treatment equals the duration of the study
cTumor Incidence expressed as number of animals with tumors/number of
animals In group
-square method
0087d
-29-
03/31/88
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(Sellakumar et al., 1969) was available only as an abstract and there were
very few details given regarding length of study, actual tumor Incidences
and statistical significance.
Twenty-six male ICR/JCL mice were fed diets containing 0.1% 3,3'-d1-
chlorobenzldlne (purity unspecified) for up to 12 months (Osanal, 1976).
Eight of the 3,3'-d1chlorobenz1d1ne-treated mice were sacrificed at 6 months
and the remaining 18 at 12 months. Of 39 control mice fed the standard
diet, 5 were sacrificed at 6 months, 21 were sacrificed at 12 months and 13
were sacrificed at 18 months. The Incidence of hepatomas In 3,3'-d1chloro-
benzldlne-treated mice was 100% at both sacrifice times (Table 6-3).
Control mice killed at 6, 12 and 18 months had hepatoma Incidences (and mean
numbers of tumors) of 0, 9.5% (two hepatomas/mouse) and 38.5% (five
hepatomas/mouse), respectively.
The data base regarding carclnogenlclty of 3,3'-d1chlorobenz1d1ne In
humans 1s limited. The available ep1dem1olog1ca1 studies are summarized as
follows.
3,3'-d1ch1orobenz1d1ne Is suspected of being a bladder carcinogen In
humans based on Us structural resemblance to the known human bladder
carcinogen, benzldlne. Based on results of an ep1dem1o1og1cal study of 207
workers exposed to 3,3'-d1ch1orobenz1d1ne during the manufacture of dyes at
the Allied Chemical Corp., Haledon, NJ (Gerarde and Gerarde, 1974), the
authors concluded that 3,3'-d1chlorobenz1d1ne does not cause bladder cancer
1n humans. Individuals Included 1n the study were those who had worked with
or come 1n contact with 3,3'-d1chlorobenz1d1ne between 1938 and 1975, and
their occupational and medical histories were examined. These were complete
medical histories or follow-up reports on 175 workers (163 living and 12
0087d -30- 05/09/88
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TABLE 6-3
Incidence of Hepatomas 1n Male ICR/JCL Mice Fed 0.1%
3,3-D1chlorobenz1d1ne 1n the D1eta
Dose
0
0
0
0.1%
0.1%
Duration of Treatment*1
(months)
6
12
18
6
12
Tumor Incidence0
(p value)
0/5
2/21
5/13
8/8 (p=0.0008)
18/18 (p=3x!0~»)
Strengths of study:
Weakness of study:
QUALITY OF EVIDENCE
Natural route of exposure
Small number of animals/group; only males tested;
purity of compound not specified; only one dose tested;
high spontaneous rate of tumor development In control.
Overall adequacy: Limited
aSource: Osanal, 1976
^Duration of the study equals the duration of treatment
°F1sher Exact test performed at SRC
0087d
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03/31/88
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deceased) having exposure to 3,3'-d1chlorobenz1d1ne ranging from 1 month to
24 years with the majority exposed for <15 years.
A similar retrospective ep1dem1olog1cal study was conducted of workers
handling 3,3'-d1chlorobenz1d1ne In a plant 1n Britain (Maclntyre, 1975).
There were no cases of bladder cancer found 1n a population of 225 workers
exposed to 3,3'-d1chlorobenz1d1ne over a period of 30 years (with the
majority exposed for less than 16 years).
A problem that arises when studying occupational exposure to 3,3'-d1-
chlorobenzldlne 1s that often workers who handle 3,3'-d1chlorobenz1d1ne are
also exposed simultaneously to other known human carcinogens such as benzl-
dlne. Gadlan (1975) studied workers at the Clayton Aniline Company who had
worked with 3,3'-d1chlorobenz1d1ne or benzldlne between 1953 and 1973. The
Incidence of urinary tract tumors In workers exposed to 3,3'-d1chlorobenz1-
dlne and benzldlne was 3/14 (2 carcinomas of bladder and 1 papllloma of
bladder), whereas no bladder tumors were found 1n 35 workers exposed to
3,3'-d1ch1orobenz1d1ne only. Gadlan (1975) concluded that although the
number of workers was small 1n this study, the findings did suggest that
3,3'-d1chlorobenz1d1ne does not cause bladder tumors 1n humans.
The three ep1dem1olog1cal studies (Gerarde and Gerarde, 1974; Gadlan,
1975; Maclntyre, 1975) discussed above have been criticized by IARC (1982)
because of a number of deficiencies. All of these studies examined
relatively small cohorts of workers (the largest cohort consisted of 225
workers) and they all had limited statistical power to detect Increases In
bladder cancer. Host of the workers Included In these three studies were
exposed to 3,3'-d1chlorobenz1d1ne for <20 years and follow-up of exposed
workers In the study by Gerarde and Gerarde (1974) was <85% complete.
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Because of the high survival rate and long latency, Increased Incidences of
bladder tumor may not be apparent 1f cohorts are not followed for a long
period of time or 1f only mortality of the cohorts Is analyzed.
An explanation of the finding that 3,3'-d1chlorobenz1d1n.e does not cause
bladder tumors 1n humans but apparently 1s capable of producing a variety of
tumors In animals was proposed by Parkes and Evans (1984), who suggested
that the levels of 3,3'-d1chlorobenz1d1ne to which workers are exposed are
sufficiently low, compared with the experimental doses used In animals, so
that no carcinogenic effect 1s observed.
6.2.3. Other Relevant Information. The cardnogenlclty of 3,3'-d1chloro-
benz1d1ne following subcutaneous administration In rats was studied by Pllss
(1959). Twenty-five female and 36 male rats received weekly subcutaneous
Injections of an 8.8% suspension of 3,3'-d1chlorobenz1d1ne In glycerol at a
dose of 120 mg 3,3'-d1ch1orobenz1d1ne/rat for the first 5 months of the
study. Because of toxic effects the dose was reduced to 20 tug/rat beginning
on the 6th month. The total 3,3'-d1chlorobenz1d1ne dose over the entire
period of the study (10-11 months) was -1.62 g/rat. The animals were
observed for life. Survival was 40 rats at 6 months, 35 rats at the time of
appearance of the first tumor (7 months) and 23 rats at 12 months. Control
animals (130 rats) were Injected with either octadecylamlne or methyl-
stearylamlne and were observed for 23 months. Of the rats surviving to the
time of the first tumor, 26 (74.3%) had tumors at different sites. Ten rats
had tumors of the Zymbal gland, five had skin tumors, six had mammary gland
tumors, seven had local subcutaneous sarcomas, two had remote tumors of the
connective tissue, two had haematopoietic system tumors and one had a
salivary gland tumor. No tumors were observed In control animals.
In another study by Pllss (1963), an unspecified number of rats received
subcutaneous Injections of 3,3'-d1chlorobenz1d1ne (15-60 mg/rat) at unsped-
0087d -33- ~ 05/09/88
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fled Intervals for 10-13 months. Fifty control rats were Injected with the
vehicle alone (sunflower seed oil or glycerol) or left untreated. Of the
3,3'-d1chlorobenz1d1ne-treated rats 74% developed tumors, with skin,
sebaceous and mammary gland tumors being observed most frequently. One
tumor was observed 1n control rats.
Both additive and synerglstlc tumor1gen1c effects were noted 1n rats
given simultaneous or sequential administration of 3,3'-d1chlorobenz1d1ne,
BNN, FANFT and 2-AAF (Ito et al., 1983). Simultaneous administration of
3,3'-d1cnlorobenz1d1ne and BBN, or 3,3'-d1chlorobenz1d1ne, 8BN and 2-AAF In
the drinking water and diet of rats resulted In a significant synerglstlc
effect of these chemicals on urinary bladder carclnogenesls over that seen
when the chemicals were administered singly. Significant additive effects
on urinary bladder carclnogenesls were seen when BBN, FANFT, 2AAF and
3,3'-d1chlorobenz1d1ne were given sequentially to rats In the diet or In
drinking water (Ito et al., 1983).
3,3'-D1chlorobenz1d1ne has also been demonstrated to function as a
transplacental carcinogen (Golub et al., 1975). A group of 24 BALB/c mice
(11 female and 13 male) was treated with five subcutaneous Injections of
3,3'-d1chlorobenz1d1ne during the last week of pregnancy (2 mg/lnjectlon,
total dose of 10 mg/mouse). A control group of 30 mice was treated with
vehicle only (0.1 ml sunflower oil). All experimental animals, Including
treated animals and offspring of treated animals, were observed over a life-
time. The Incidence of tumors 1n the offspring of 3,3'-d1chlorobenz1d1ne-
treated mice was 13/24 compared with a tumor Incidence of 6/30 In the
offspring of controls. The Incidence of lymphold leukemlas was signifi-
cantly different between offspring of 3,3'-d1chlorobenz1d1ne-treated mice
(Incidence of 7/24) and control offspring (Incidence of 0/30).
0087d -34- " 05/09/88
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An In vitro method using degranulatlon of mlcrosomes has been shown to
successfully predict the carcinogenic nature of a number of compounds,
Including 3,3'-d1chlorobenz1d1ne (Jagota and Dan1, 1985; Gupta and Oan1,
1986). In this assay, rat Hver mlcrosomes are prepared by using a method
that uses sedimentation at low G force (10,000 g). Mlcrosomes prepared 1n
this manner contain a large number of rlbosomes/unlt area, and Incubation of
these mlcrosomes with known carcinogens has been shown to result In
significant degranulatlon of the mlcrosome. Incubation of the mlcrosomes
with noncarclnogens does not result 1n a high degree of degranulatlon. Of a
number of carcinogens tested, 3,3'-d1chlorobenz1d1ne was demonstrated to
cause a high percentage of mlcrosomal degranulatlon.
3,3'-D1chlorobenz1d1ne has also been demonstrated to be capable of
producing cell transformation' ln_ vitro 1n high-passage rat embryo cell
cultures (Freeman et al., 1973). 3,3'-D1chlorobenz1d1ne was active In this
assay at a concentration of 5.0 jig/mi but not at 1.0 vg/ma.
6.3. MUTAGENICITY
3,3'-D1chlorobenz1d1ne has been tested for mutagenlcHy using various
strains of Salmonella typh1mur1um (Garner, 1975; Lazear and Louie, 1977;
Anderson and Styles, 1978; Reid et al., 1984; Iba, 1986; VHhayathll et al.,
1983; Savard and Josephy, 1986; Prlval et al., 1984; OeFrance et al., 1986;
Commoner, 1976; Gentile et al., 1985). The results shown 1n Table 6-4
Indicate that 3,3'-d1chlorobenz1d1ne acts as a mutagen towards Salmonella
typh1mur1um. both 1n the presence and absence of metabolic activation (I.e.,
various liver S-9 preparations). The direct mutagenlc activity of 3,3'-d1-
chlorobenzldlne has, however, been demonstrated by several Investigators
(Garner, 1975; Lazear and Louie, 1977; DeFrance et al., 1986) to be In-
creased anywhere from 3- to 50-fold by the addition of liver metabolizing
0087d -35- ~ 05/09/88
-------
TABLE 6-4
Hutagentctty Testing of 3,3'-Dlchlorobenz1dlne
00
a.
Assay
Reverse
nutation
Reverse
nutation
Reverse
nutation
CO
' Reverse
nutation
' Reverse
nutation
Reverse
nutation
Reverse
nutation
Reverse
nutation
Indicator/
Organism
Salmonella
typhlnurlun
TA1S38
S. typhlnurlun
TA98
S. typhlnurlun
TA1535. TA1538.
TA9B. TA100
S. typhlnurlun
TA98, TA1538
S. typhlnurlun
TA98
S. typhlmurlun
TA98
S. typhlmurlun
TA98. TA98/
1.8-DNP6
S. typhlrourlun
TA98
Compound
and/or
Purity
purified
DCB
technical
grade DCB
sulfate
DCB/NR
DCB-2HC1/
NR
NR/NR
DCB-2HC1/
>98X
NR/NR
NR/NR
NR/>99X
NR/NR
Application
plate
Incorporation
plate
Incorporation
plate
Incorporation
plate
Incorporation
plate
Incorporation
plate
Incorporation
plate
Incorporation
plate
Incorporation
plate
Incorporation
plate
Incorporation
Concentration
or Dose
SO or 100
tig/plate
50 or 100
tig/plate
SO or 100 -
wg/plate
SO or 100
tig/plate
50-500
vg/plate
5-20 yg/plate
NR
10 tig/plate
1-300
nnol/plate
1-10 tig/plate
Activating Systen
fS-9 (rat liver)
±S-9 (rat liver)
»S-9 (mouse lUer)
iS-9 (nouse liver)
»S-9
»S-9
»S-9
»S-9
»S-9
.S-9
Response Comment
«•/» Direct nutagenlc activ-
ity of compound was In-
creased 50-fold by
»/» addition of liver S-9
preparation
*A Direct nutagenlc activ-
ity of compound was In-
creased by -3- to 6-fold
»A by addition of liver S-9
fraction
* NC
«• NC
«• Pretreatment of rats
with phenobarbltal en-
hanced S-9-catalyzed
mutagenlctty of DCB by
2.3-fold
» S-9 prepared from Aro-
clor-lnduced rat liver
»/» S-9 prepared from
Syrian golden hamsters
* Liver S-9 preparation
from phenobarbltone pre-
Reference
Garner. 1975
Lazear and
Louie. 1977
Anderson and
Styles. 1978
Reid et al..
1984
Iba. 1986
Vlthoyathll
et al.. 1983
Savand and
Josephy. 1986
Booth et al..
1980
treated rats was more
effective In converting
DCB to mutagen then
liver S-9 from Aroclor
1254 pretreated rats
03
00
-------
o
o
GO
TABIF 6-4 (cont.)
Assay
Reverse
mutation
Reverse
nutation
Reverse
nutation
Reverse
mutation
SCI
Unscheduled
DNA synthesis
Indicator/
Organlsn
S. typhlmurlum
TA98
S. typhlnurlun
TA98
S. typhlmurlum
TA9B
S. typhlnnirlua
TA98
S. typhlmurlua
TA1S3B
Bloom syndrome
0-lyraphoblastotd
cell line
HeLa cells
Compound
and/or
Purity
OCB-2HC1/
NR
NR/NR
NR/NR
NR/NR
NR/NR
NR/NR
NR/NR
Application
plate
Incorporation
plate
Incorporation
plate
Incorporation
plate
Incorporation
plate
Incorporation
\jn vitro cell
culture
Jn vitro cell
culture
Concentration
or Dose
10-30
nmol/plate
1-10
nmol/plate
0-?00 yg/plate
10 vg/plate
0.5-10
iig/plate
1.7xlO~« to
1.3xlfl-» N
10"« to
10"» N
Activating System
S-9 (rat liver)
»S-9 (hamster
liver)
»S-9 (hamster
liver)
»S-9 (hamster
liver)
»S-9 (from Aroclor
1254-tnduced rat
liver)
»S-9 (rat liver)
»S-9 (rat liver)
Response Comment
» NC
Reference
Prlval
et al..
»/» Direct mutagenlc acttv- Oefrance
Ity of compound was In- et al.,
creased -4-fold by addi-
tion of liver S-9 fraction
* NC
f Results were similar for
S-9 obtained from livers
of Shlstosome-lnfested
hamsters control hamsters
«• Increasing concentra-
tions of Aroclor 1254-
tnduced rat liver S-9
decreased DCB mutagenlclty
»/» SCE/cell were greater
In presence of S-9
f NC
Commoner
1976
Gentile
et al..
Gentile
et al..
1984
1986
•
1985
1985
Shlralshl.
1986
Martin
et al..
1978
NC = No comment; NR = not reported
OS
-------
enzymes. Iba (1986) reported that epoxldatlon of 3,3'-d1chlorobenz1d1ne to
form an arene oxide may be Involved In the activation of the compound.
The type of Inducer of liver enzymes used before the preparation of S-9
also appears to be Important when studying the mutagenlcHy of 3,3'-d1-
chlorobenzldlne. It has been shown that Aroclor-1254 Is a relatively poor
Inducer of the enzyme(s) responsible for 3,3'-d1chlorobenz1d1ne activation
(Garner, 1980; Booth et al., 1980) and In fact Increasing concentrations of
liver S-9 from Aroclor !254-1nduced rats have been shown to actually
decrease the mutagenlcHy of 3t3'-d1chlorobenz1d1ne (Gentile et al., 1985).
3,3'-D1chlorobenz1d1ne has also been demonstrated to be active In an in
vitro assay measuring sister chromatld exchange 1n a B-lymphoblasto1d cell
line (Shlralshl, 1986) and In an J£ vitro assay measuring unscheduled ONA
synthesis 1n HeLa cells (Martin et al., 1978).
6.4. TERATOGENICITY
Shabad et al. (1972) studied the effects of 3,3'-d1chlorobenz1d1ne
administration to pregnant mice (during last week of pregnancy) on the
embryonic kidney. Pregnant Balb/c mice were treated by subcutaneous
Injection with 8-10 mg 3,3'-d1chlorobenz1d1ne In sunflower oil and then
fragments of embryonic kidney were explanted 1n organ culture. Embryonic
kidney cultures obtained from 3,3'-d1chlorobenz1d1ne-exposed mice were shown
to have a longer survival time than control cultures and 1n addition there
was an Induction of hyperplasla of epithelial structures In the cultures
obtained from 3,3'-d1chlorobenz1d1ne-exposed mice.
6.5. OTHER REPRODUCTIVE EFFECTS
Pertinent data regarding other reproductive effects of 3,3'-d1chloro-
benzldlne were not located 1n the available literature cited 1n Appendix A.
0087d -38- 05/09/88
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&.(>. SUMMARY
Pertinent data regarding the systemic toxldty of 3,3'-d1chlorobenz1d1ne
following either subchronlc or chronic Inhalation exposure 1n humans and
animals were not located 1n the available literature. Stula et al. (1978)
Indicated that beagle dogs exposed to 3,3'-d1chlorobenz1d1ne orally for
periods up to 7.1 years showed signs of liver toxldty 1n the form of
elevated SGPT activities. The oral L05Q of 3,3'-d1chlorobenz1d1ne In rats
has been reported to be between ~4 and 7 g/kg bw (ACGIH, 1986).
Pertinent data regarding the cardnogenlcHy of 3,3'-d1chlorobenz1d1ne
following Inhalation exposure In humans and animals were not located In the
available literature cited In Appendix A. Oral administration of 3,3'-d1-
chlorobenzldlne has been shown to produce a variety of tumors 1n rats
(Pllss, 1959; Stula et al., 1975; GMswold et al., 1968), urinary bladder
and liver tumors In dogs {Stula et al., 1978) and hamsters (Sellakumar et
al., 1969), and hepatomas 1n mice (Osanal, 1976). Subcutaneous administra-
tion of 3,3'-d1chlorobenz1d1ne has also been demonstrated to produce tumors
In rats (Pllss, 1959, 1963).
Both additive and synerglstlc tumorlgenlc effects were noted In rats
following simultaneous or sequential administration of low levels of
3,3'-d1chlorobenz1d1ne along with low levels of other carcinogens (such as
BBN, FANFT and 2-AAF) (Ito et al., 1983). 3.3'-d1chlorobenz1d1ne has also
been demonstrated to function as a transplacental carcinogen In mice (Golub
et al., 1975).
3,3'-D1chlorobenz1d1ne has been suspected of being a human carcinogen
because of Us carcinogenic effects 1n animals and because 1t resembles the
known human bladder carcinogen benzldlne. Evidence from three epidemic-
logical studies (Gerarde and Gerarde, 1974; Maclntyre, 1975; Gadlan, 1975),
0087d -39- 05/09/88
-------
however. Indicates that 3,3'-d1chlorobenz1d1ne Is not a bladder carcinogen
1n humans.
3,3'-D1chlorobenz1d1ne has been demonstrated to be mutagenlc towards
Salmonella typhlmurlum In the Ames assay, both with and without metabolic
activation. Metabolic activation (I.e., presence of liver S-9) has, however,
been demonstrated to Increase the mutagenldty of 3,3'-d1chlorobenz1d1ne
from 3- to 50-fold (Garner, 1975; Lazear and Louie, 1977; OeFrance et al.,
1986). 3,3'-D1chlorobenz1d1ne has also been demonstrated to be active In in
vitro assays measuring unscheduled ONA synthesis (Martin et al., 1978) and
sister chromatld exchange (Shlralshl, 1986).
Pertinent data regarding the tetratogenlclty of 3,3'-d1chlorobenz1d1ne
/"•
were not located 1n the available literature. One study (Shabad et al.,
1972) demonstrated that transplacental exposure of mice to 3,3'-d1chloro-
benzldlne had effects on the growth of embryonic kidney cells 1n culture.
0087d -40- 05/09/88
-------
7. EXISTING GUIDELINES AND STANDARDS
7.1. HUMAN
Because ACGIH (1987) lists 3,3'-d1chlorobenz1d1ne as a suspected human
carcinogen, there 1s no TLV-TWA for 3,3'-d1chlorobenz1d1ne. 3,3'-D1chloro-
benzldlne 1s suspected of being a human carcinogen because of Its structural
resemblance to benzldlne, the known human bladder carcinogen; and because
3,3'-d1chlorobenz1d1ne has been demonstrated to be carcinogenic In experi-
mental animals {ACGIH, 1986).
7.2. AQUATIC
The data base for the aquatic toxldty of 3,3'-d1ch1orobenz1d1ne 1s
limited and guidelines and standards for the protection of aquatic organisms
were not located In the available literature cited In Appendix A.
0087d -41- 04/01/88
-------
8. RISK ASSESSMENT
8.1. CARCINOGENICITY
8.1.1. Inhalation. Pertinent data regarding the carclnogenlclty of
3t3'-d1chlorobenz1d1ne following Inhalation exposure 1n either animals or
humans were not located In the available literature cited In Appendix A.
8.1.2. Oral. In a study by PUss (1959), 50 rats (15 female and 35 male)
were fed 3,3'-d1ch1orobenz1d1ne In a paste which provided a dose of 10-20 mg
3,3'-d1chlorobenz1d1ne/day. The dose was administered 6 days/week for 12
months (total dose of 4.5 g 3t3'-d1chlorobenz1d1ne/rat) and the rats were
observed for a lifetime. Twenty-three of the rats developed a variety of
tumors.
In rats fed 3,3'-d1chlorobenz1d1ne 1n the diet (1000 ppm) for 349-353
days (Stula et al., 1975), significantly Increased Incidences of granulo-
cytlc leukemlas, mammary adenocardnomas and Zymbal gland carcinomas were
found 1n males compared with controls. In female rats treated with same
regimen, the Incidence of mammary adenocardnomas was significantly
Increased as compared with controls.
No mammary tumors were found 1n female rats treated by gastric Intuba-
tion with 10 doses of 3,3'-d1chlorobenz1d1ne over a 30-day period (total
dose of 300 mg/rat), whereas the Incidence of mammary tumors In OMBA-treated
positive controls In the same experiment was 100% (GHswold et al., 1968).
Significantly Increased Incidences of bladder carcinomas and hepato-
cellular carcinomas, as compared with controls, were found In beagle dogs
given a dally oral dose of 100 mg 3,3'-d1chlorobenz1d1ne In capsular form 3
times/ week for 6 weeks followed by 5 times/week for up to an additional 7
years (Stula et al., 1978).
0087d -42- 04/01/88
-------
No evidence of cardnogenldty was found 1n hamsters fed a diet contain-
ing 0.1% (w/w) 3,3'-d1chlorobenz1d1ne (60 mg/hamster/week) over the course
of a lifetime (Safflottl et al.. 1967). Hamsters fed a diet containing 0.3%
(w/w) 3,3'-d1chlorobenz1d1ne (dosing schedule unspecified), however,
developed transitional cell bladder carcinomas and some liver-cell and
cholanglomatous tumors (Sellakumar et al., 1969). Individual tumor
Incidences were not given.
In mice fed 3,3'-d1chlorobenz1d1ne In the diet (0.1% w/w) for up to 12
months (Osanal, 1976), the Incidence of hepatomas was 100% at both sacrifice
times (6 and 12 months), whereas control mice killed at 6, 12 and 18 months
had hepatoma Incidences of 0, 9.5 and 38.5%, respectively.
8.1.3. Other Routes. PUss (1959) reported that In rats receiving weekly
subcutaneous doses of 3,3'-d1chlorobenz1d1ne suspended 1n glycerol for 10-11
months (total dose of 1.62 g/rat), 74.3% had tumors at different sites. No
tumors were observed In control rats.
In another study, Pllss (1963) found that 74% of rats receiving subcuta-
neous Injections of 3,3'-d1chlorobenz1d1ne (15-60 mg/rat) at unspecified
Intervals for 10-13 months developed tumors, with skin, sebaceous and
mammary gland tumors being observed most frequently. One tumor was observed
1n control rats.
Three ep1dem1olog1cal studies of workers occupatlonally exposed to low
levels of 3,3'-d1chlorobenz1d1ne are Inadequate for assessment of human
carc1nogen1dty of 3,3'-d1chlorobenz1d1ne due to deficiencies such as:
small cohorts, limited statistical power and short exposure periods (Gerarde
and Gerarde, 1974; Haclntyre, 1975; Gadlan, 1975).
8.1.4. Weight of Evidence. There 1s sufficient evidence from several
studies (PUss, 1959, 1963; Stula et al., 1975, 1978; Sellakumar et al.,
1969; Osanal, 1976) that 3,3'-d1chlorobenz1d1ne functions as a carcinogen 1n
0087d -43- 05/09/88
-------
animals. Data from three ep1dem1olog1cal studies (Gerarde and Gerarde,
1974; Maclntyre, 1975; Gadlan, 1975) have several methodological limitations
(see Section 6.2.2.). hence there 1s Inadequate human evidence for cardno-
genldty. Because there 1s sufficient evidence In animals and Inadequate
evidence 1n humans, 3,3'-d1chlorobenz1d1ne 1s categorized 1n EPA Group B2 -
probable human carcinogen (U.S. EPA, 1986b).
8.1.5. Quantitative Risk Estimates.
8.1.5.1. INHALATION — Because pertinent data regarding the cardno-
genldty of 3,3'-d1chlorobenz1d1ne following Inhalation exposure were not
located 1n the available literature, an Inhalation q * for 3,3'-d1chloro-
benzldlne was not calculated.
8.1.5.2. ORAL — The study chosen for the derivation of the oral
q.j* was that of Stula et al. (1975), 1n which female rats fed 3,3'-d1-
chlorobenzldlne 1n the diet (1000 ppm) over the course of a lifetime had a
statistically significant Increased Incidence of mammary adenocarclnomas
compared with controls. This Incidence of mammary adenocarclnomas 1n female
rats (59.1%) was larger than the Incidences of granulocytlc leukemia
(20.5%), mammary adenocardnoma (15.9%) and Zymbal gland carcinoma (18.2%)
observed In male rats In the same study. The study by Stula et al. (1978)
on the cardnogenlcty of 3,3'-d1chlorobenz1d1ne 1n beagle dogs was not
considered for derivation of a q * because of the small number of dogs
(six) used In the study. The study by Osanal (1976) of the effects of
3,3'-d1chlorobenz1d1ne exposure on the hepatoma Incidence In mice was not
considered for q,* development because of the small number of control
animals and because the duration of exposure (6 or 12 months) was fairly
short compared with the Hfespan of the animal (2 years).
The oral q,* for 3,3'-d1chlorobenz1d1ne was calculated using the
multistage model developed by Howe and Crump (1982). An unadjusted (animal)
0087d -44- 05/09/88
-------
q,* of 2.3xlO~a (mg/kg/day)"1 was calculated using the Incidences of
mammary adenocardnomas 1n female rats fed either 0 or 1000 ppm 3,3'-d1-
chlorobenzldlne 1n the diet over the course of a lifetime (range of days on
test: 143-488). The exposure level of 1000 ppm 3,3'-d1chlorobenz1d1ne 1n
the diet was converted to a dose of 50 mg/kg/day using a food factor of 0.05
for rats (U.S. EPA, 1986d). Multiplication of the unadjusted q.,* by the
cube root of the ratio of human body weight (70 kg) to rat body weight (0.35
kg) (U.S. EPA, 1986d) and by the cube .of the ratio of the llfespan of the
rat (730 days) (U.S. EPA, 1986d) to the average length of the experiment
(349 days) results 1n a human q * of 1.2 (mg/kg/day)'1.
To derive the concentration of 3,3'-d1chlorobenz1d1ne In the drinking
water associated with an Increased lifetime risk of cancer at a risk level
of 10~5, the risk level was divided by the human q * of 1.2 (mg/kg/
day)"1 to give a dose of 8.3x10"' mg/kg/day. Multiplying this value by
the human body weight (70 kg) and dividing by the amount of water consumed
by an Individual each day (2 l) (U.S. EPA, 1986d) results In a concentra-
tion of 2.9x10"* mg 3,3'-d1chlorobenz1d1ne/j. 1n the drinking water asso-
ciated with a risk level of 10"5. Concentrations of 3,3'-d1chlorobenz1d1ne
1n the drinking water associated with risk levels of 10"' and 10~7 are
2.9xlO"s and 2.9x10"* mg 3,3'-d1chlorobenz1d1ne/l, respectively.
8.2. SYSTEMIC TOXICITY
8.2.1. Inhalation Exposures. Pertinent data regarding the systemic
toxiclty of 3,3'-d1chlorobenz1d1ne following subchronlc or chronic Inhala-
tion exposure In either animals or humans were not located 1n the available
literature cited 1n Appendix A. This precluded the derivation of Inhalation
RfDs. Furthermore, because there 1s sufficient evidence that 3,3'-d1chloro-
benzldlne Is a carcinogen, 1t 1s not appropriate to derive an RfD.
0087d -45- 06/06/88
-------
8.2.2. Oral Exposures. Dogs given an oral dose of 3,3'-d1chlorobenz1d1ne
(100 mg/day) over an extended period of time (I.e., up to 7.1 years) showed
signs of liver toxlclty 1n the form of elevated SGPT activities (Stula et
al.t 1978). Because 3,3'-d1chlorobenz1d1ne has been demonstrated to be
carcinogenic, subchronlc and chronic oral RfDs were not derived.
0087d -46- 03/31/88
-------
9. REPORTABLE QUANTITIES
9.1. BASED ON SYSTEHIC TOXICITY
In a previous determination (U.S. EPA, 1986c), an RQ for 3,3'-d1chloro-
benzldlne based on systemic toxldty was not derived because the data were
considered Insufficient to derive an RQ. Reevaluatlon of the data presented
In the study of 3,3'-d1chlorobenz1d1ne carclnogenlcUy 1n beagle dogs (Stula
et al., 1978) has led to the development of the RQ for 3,3'-d1chlorobenz1-
dlne presented here. Since the study by Stula et al. (1978) 1s the only
chronic study that gives toxldty Information on 3,3'-d1chlorobenz1d1ne, It
1s the only study considered for RQ development. This study 1s discussed In
Chapter 6 and 1s summarized 1n Table 9-1.
The only effect considered for RQ development 1s elevated SGPT activi-
ties In dogs (see Table 9-1), which occurred at an equivalent human dose of
3.8 mg/kg/day. Multiplication of this dose by the human body weight (70 kg)
gives an MED of 266 mg/day, which corresponds to an RV. of 1.9 (RV. =
-1.5 log MED > 5.5). An RV of 6 was assigned to the effect of elevated
SGPT, which represents clinical evidence of hepatocellular necrosis.
Multiplication of the RV (1.9) by the RV (6) gives a CS of 11. The RQ
associated with this CS 1s 1000 (Tables 9-2 and 9-3).
The basis for the derivation of this RQ based on systemic toxldty of
3,3'-d1chlorobenz1d1ne 1s weak. The study by Stula et al. (1978) 1s the
only study with chronic toxldty Information on 3,3'-d1chlorobenz1d1ne, and
only one dose level was used In this study. This dose level and the result-
Ing effect of elevated SGPT activity therefore define a free-standing LOAEL.
Because 3,3'-d1chlorobenz1d1ne has been demonstrated to be carcinogenic 1n
animals, the more conservative RQ of 10 derived from the carclnogenlcUy
data (see section 9.2) 1s recommended for the 3,3'-d1chlorobenz1d1ne.
0087d -47- 04/01/88
-------
TABLE 9-1
Oral ToxIcHy Summary for 3,3'-D1chlorobenz1d1ne Using the Female Beagle3
No. at
Start
6
Average
Weight
(kg)
9.73C
Vehicle/
Physical Purity
State
gelatin 100X
capsule
Exposure
100 mg/day. 3
times/week for 6
weeks then 5
times/week for an
additional 7 years
(total 7.1 years)
TWA - 71 mg/day d
Transformed
Animal Dose
(mg/kg/day)
7.30
Equivalent
Human Ooseb
(mg/kg/day)
3.8
Response
elevated SGPT
activities
^Source: Stula et al., 1978
^Calculated by multiplying the animal transformed dose by the cube root of the ratio of the animal body
weight to the human body weight (70 kg)
Determined from body weight data provided In the study
>dTUA dose calculated as follows:
(100 mg/day x 3 davs/7 days x 6 weeks) » (100 mg/dav x 5 days/7 days x 364 weeks)
370 weeks
e71 mg/day * 9.73 kg = 7.3 mg/kg/day
roq/day
-------
TABLE 9-2
Oral Composite Score for 3,3'-D1chlorobenz1d1ne Using the Dog*
Animal Dose
(nuj/kg/day)
Chronic
Human MED
(mg/day)
RVd
Effect
CS
RQ
7.3
266
1.87 elevated SGPT
activities
11.2 1000
*Source: Stula et al., 1978
0087d
-49-
03/31/88
-------
TABLE 9-3
3,3'-D1chlorobenz1d1ne
Minimum Effective Dose (MED) and Reportable Quantity (RQ)
Route:
Dose*:
Effect:
Reference:
RVd:
RVe:
Composite Score:
RQ:
oral
266 mg/day
elevated SGPT activities
Stula et al.. 1978
1.9
6
11
1000
0087d
-50-
03/31/88
-------
9.2. BASED ON CARCINOGENICITY
There were no data available regarding the carclnogenlcHy of 3,3'-d1-
chlorobenzldlne following Inhalation exposure. Studies considered for the
development of an RQ based on carc1nogen1c1ty were presented In Section
6.2.2. and are summarized In Tables 6-1, 6-2 and 6-3. Six female dogs given
100 mg 3,3'-d1chlorobenz1d1ne/day, 3 times/week for the first 6 weeks and
then 5 times/week for up to an additional 7 years (Stula et al., 1978)
developed papillary transitional cell carcinomas of the urinary bladder and
hepatocellular carcinomas. This study was not considered for RQ development
because of the small numbers of animals used. In another study, male and
female rats were fed 3,3'-d1chlorobenz1d1ne (1000 ppm) 1n the diet over the
course of a lifetime (range of days on test: 143-488; average days on test:
349 days) (Stula et al., 1975). 3,3'-D1chlorobenz1d1ne-exposed female rats
had a statistically significant Increased Incidence of mammary adenocarclno-
mas compared with controls, and this study was chosen for q,* and RQ
development. In another study considered for RQ development, mice were
exposed to 3,3'-d1chlorobenz1d1ne 1n the diet (0.1% w/w) for 6 or 12 months
(Osanal, 1976). The Incidence of hepatomas In the 3,3'-d1chlorobenz1d1ne-
treated mice was 100% In both groups. This study was not used for q * or
RQ development because of the small number of control animals (39 were used)
and because the length of exposure (6 or 12 months) was relatively short
compared to the lifetime of the animal.
3,3'-D1chlorobenz1d1ne has been shown to be carcinogenic 1n a number of
animal studies (see Section 6.2.), but three ep1dem1olog1cal studies
(Gerarde and Gerarde, 1974; Maclntyre, 1975; Gadlan, 1975) are Inadequate
for determining whether 3,3'-d1chlorobenz1d1ne causes cancer In humans.
Therefore, 3,3'-d1chlorobenz1d1ne Is categorized 1n EPA Group B2 - probable
human carcinogen (U.S. EPA, 1986b).
0087d -51- 05/09/88
-------
The unadjusted 1/ED,Q for 3,3'-d1chlorobenz1d1ne based on the study of
Stula et al. (1975) Is 0.15626 (mg/kg/day)'1 and was derived using the
multistage model developed by Howe and Crump (1982) (Table 9-4). Multipli-
cation of this unadjusted 1/ED,Q by the cube root of the ratio of human
body weight (70 kg) to rat body weight (0.35 kg) and by the cube of the
ratio of the Hfespan of the animal (730 days) to the average length of the
experiment (349 days) results 1n an F Factor for humans of 8.4 (mg/kg/
day)'1. This F Factor 1s the same as the one derived by U.S. EPA (1986c)
and places 3,3'-d1chlorobenz1d1ne 1n Potency Group 2. Because 3,3'-d1-
chlorobenzldlne 1s categorized 1n EPA Group B2 and Potency Group 2 the
compound has a MEDIUM hazard ranking under CERCLA. A medium hazard ranking
Is associated with an RQ of 10.
0087d -52- 03/31/88
-------
TABLE 9-4
Derivation of Potency Factor (F) for 3,3'-D1chlorobenz1d1ne
Reference:
Exposure Route:
Species:
Strain:
Sex:
Vehicle State:
Body Weight:
Duration of Treatment:
Duration of Study:
UFespan of Animal:
Target Organ:
Tumor Type:
Experimental Dose or Exposures:
Transformed Dose:
Tumor Incidences:
Unadjusted 1/ED-|Q:
Adjusted 1/ED10:
(F factor)
Stula et al.. 1975
oral, diet
rat
CHR-CD
female
3,3'-d1chlorobenz1d1ne mixed In standard
diet containing 1% added corn oil
0.35 kg
349 days
349 days
730 days
mammary gland
adenocardnoma
0 ppm
0 mg/kg/day
3/44
0.15626 (mg/kg/day)"1
8.36273 (mg/kg/day)'1
1000 ppm
50 mg/kg/day
26/44
0087d
-53-
03/31/88
-------
10. REFERENCES
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0087d -58- 03/31/88
-------
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0087d -59- 03/31/88
-------
Kellner, H.M., et al. 1973. Animal studies on the kinetics of benzldlne
and 3,3'-d1chlorobenz1d1ne. Arch. Toxlcol. 31: 61. (Cited In U.S. EPA,
1980)
Lapp, T.W., T.L. Ferguson, H. Gadberry, F. Hoffmelster and F. Hopkins.
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0087d -60- 03/31/88
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0087d -61- 03/31/88
-------
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placenta! effect of some chemical compounds on organ cultures of embryonic
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0087d -62- 03/31/88
-------
Shrlner, C.R., J.S. Drury, A.S. Hammons, L.E. Towlll, E.B. Lewis and D.M.
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Benzldlne. Oak Ridge Natl. Lab., Oak Ridge, TN. EPA-600/1-78-024. 125 p.
S1kka, H.C., H.T. Appleton and S. Banerjee. 1978. Fate of 3,3'-d1chloro-
benzldlne 1n aquatic environments. EPA-600/3-78-068.
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p. 569.
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Experimental neoplasla In rats from oral administration of 3,3'-d1chloro-
benzldlne, 4,4'-methylene-b1s(2-chloroan1l1ne) and 4,4'-methylene-b1s
(2-methylan1l1ne). Toxlcol. Appl. Pharmacol. 31: 159-175.
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0087d -63- 03/31/88
-------
Troll, n.d. Personal communication. (Cited 1n Gerarde and Gerarde, 1974)
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TSCA Inventory. OPTS, CID, U.S. EPA, Washington, DC.
U.S. EPA. 1980. Guidelines and Methodology Used In the Preparation of
Health Effect Assessment Chapters of the Consent Decree Water Criteria
Documents. Federal Register. 45(31): 79347-79357.
U.S. EPA. 1981. TreatabllHy Manual I. TreatabllHy Data. U.S. EPA,
Washington, DC. EPA-600/2-82-601A.
U.S. EPA 1984. Methodology and Guidelines for Reportable Quantity Deter-
minations Based on Chronic Toxldty Data. Prepared by the Office of Health
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Cincinnati, OH for the Office of Solid Waste and Emergency Response, Wash-
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U.S. EPA. 1986a. Methodology for Evaluating Carclnogenldty In Support of
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the Office of Health and Environmental Assessment. Carcinogen Assessment
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U.S. EPA. 1986b. Guidelines for Carcinogen Risk Assessment. Federal
Register. 51(185): 33992-34003.
0087d -64- 06/06/88
-------
U.S. EPA. 1986c. Evaluation of the Potential CarclnogenlcUy of 3,3'-D1-
chlorobenzldlne. Prepared by the Carcinogen Assessment Group, Office of
Water Health and Environmental Assessment, Washington, DC for the Office of
Emergency and Remedial Response. Office of Solid Waste and Emergency
Response. OHEA-C-073-81.
U.S. EPA. 1986d. Reference Values for Risk Assessment. Prepared by the
Office of Health and Environmental Assessment, Environmental Criteria and
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DC.
U.S. EPA. 1987a. Graphical Exposure Modeling System (GEHS). Personal
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U.S. EPA. 1987b. STORET Hater Quality Data Base. Online: Oct. 1987.
USITC (U.S. International Trade Commission). 1984. Imports of Benzenold
Chemicals and Products. 1983. USITC Publ. 1548, Washington, DC. p. 15.
Vlthayathll, A.J., C. HcClure and J.W. Myers, 1983. Salmonella/mlcrosome
multiple Indicator mutagenlclty test. Mutat. Res. 121: 33-37.
0087d -65- 06/06/88
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APPENDIX A
LITERATURE SEARCHED
This HEED 1s based on data Identified by computerized literature
searches of the following:
CHEHLINE
TSCATS
CASR online (U.S. EPA Chemical Activities Status Report)
TOXLINE
TOXLIT
TOXLIT 65
RTECS
OHM TADS
STORET
SRC Environmental Fate Data Bases
SANSS
AQUIRE
TSCAPP
NTIS
Federal Register
CAS ONLINE (Chemistry and Aquatic)
HSOB
These searches were conducted In October 1987, and the following secondary
sources were reviewed:
ACGIH (American Conference of Governmental Industrial Hyg1en1sts).
1986. Documentation of the Threshold Limit Values and Biological
Exposure Indices, 5th ed. Cincinnati, OH.
ACGIH (American Conference of Governmental Industrial Hyg1en1sts).
1987. TLVs: Threshold Limit Values for Chemical Substances In the
Work Environment adopted by ACGIH with Intended Changes for
1987-1988. Cincinnati, OH. 114 p.
Clayton, G.D. and F.E. Clayton, Ed. 1981. Patty's Industrial
Hygiene and Toxicology, 3rd rev. ed., Vol. 2A. John Wiley and
Sons, NY. 2878 p.
Clayton, G.D. and F.E. Clayton, Ed. 1981. Patty's Industrial
Hygiene and Toxicology, 3rd rev. ed.. Vol. 2B. John Wiley and
Sons, NY. p. 2879-3816.
Clayton, G.D. and F.E. Clayton, Ed. 1982. Patty's Industrial
Hygiene and Toxicology, 3rd rev. ed., Vol. 2C. John Wiley and
Sons, NY. p. 3817-5112.
0087d -66- 03/31/88
-------
Grayson, M. and D. Eckroth, Ed. 1978-1984. K1rk-0thmer Encyclo-
pedia of Chemical Technology, 3rd ed. John Wiley and Sons, NY. 23
Volumes.
Hamilton, A. and H.L. Hardy. 1974. Industrial Toxicology, 3rd ed.
Publishing Sciences Group, Inc., Littleton, MA. 575 p.
IARC (International Agency for Research on Cancer). IARC Mono-
graphs on the Evaluation of Carcinogenic Risk of Chemicals to
Humans. IARC, WHO, Lyons, France.
Jaber, H.M., W.R. Mabey, A.T. L1eu, T.W. Chou and H.L. Johnson.
1984. Data acquisition for environmental transport and fate
screening for compounds of Interest to the Office of Solid Waste.
EPA 600/6-84-010. NTIS PB84-243906. SRI International, Menlo
Park, CA.
NTP (National Toxicology Program). 1987. Toxicology Research and
Testing Program. Chemicals on Standard Protocol. Management
Status.
Ouellette, R.P. and J.A. King. 1977. Chemical Week Pesticide
Register. McGraw-Hill Book Co., NY.
Sax, I.N. 1984. Dangerous Properties of Industrial Materials, 6th
ed. Van Nostrand Relnhold Co.. NY.
SRI (Stanford Research Institute). 1987. Directory of Chemical
Producers. Menlo Park, CA.
U.S. EPA. 1986. Report on Status Report In the Special Review
' Program, Registration Standards Program and the Data Call 1n
Programs. Registration Standards and the Data Call 1n Programs.
Office of Pesticide Programs, Washington, DC.
USITC (U.S. International Trade Commission). 1986. Synthetic
Organic Chemicals. U.S. Production and Sales, 1985, USITC Publ.
1892, Washington, DC.
Verschueren, K. 1983. Handbook of Environmental Data on Organic
Chemicals, 2nd ed. Van Nostrand Relnhold Co., NY.
Worthing, C.R. and S.B. Walker, Ed. 1983. The Pesticide Manual.
British Crop Protection Council. 695 p.
Wlndholz, M., Ed. 1983. The Merck Index, 10th ed. Merck and Co.,
Inc., Rahway, NJ.
0087d -67- 03/31/88
-------
In addition, approximately 30 compendia of aquatic toxldty data were
reviewed, Including the following:
Battelle's Columbus Laboratories. 1971. Water Quality Criteria
Data Book. Volume 3. Effects of Chemicals on Aquatic Life.
Selected Data from the Literature through 1968. Prepared for the
U.S. EPA under Contract No. 68-01-0007. Washington, DC.
Johnson, W.W. and M.T. Flnley. 1980. Handbook of Acute Toxldty
of Chemicals to F1sh and Aquatic Invertebrates. Summaries of
Toxldty Tests Conducted at Columbia National Fisheries Research
Laboratory. 1965-1978. U.S. Dept. Interior, Fish and Wildlife
Serv. Res. Publ. 137, Washington, DC.
McKee, J.E. and H.W. Wolf. 1963. Water Quality Criteria, 2nd ed.
Prepared for the Resources Agency of California, State Water
Quality Control Board. Publ. No. 3-A.
Plmental, D. 1971. Ecological Effects of Pesticides on Non-Target
Species. Prepared for the U.S. EPA, Washington, DC. PB-269605.
Schneider, B.A. 1979. Toxicology Handbook. Mammalian and Aquatic
Data. Book 1: Toxicology Data. Office of Pesticide Programs, U.S.
EPA, Washington, DC. EPA 540/9-79-003. NTIS PB 80-196876.
0087d -68- 03/31/88
-------
APPENDIX B
Cancer Data Sheet for Derivation of q-j*
Compound: 3,3'-D1chlorobenz1d1ne
Reference: Stula et al., 1975
Spec1es/strain/sex: rat/Chr-CD/female
Route/vehicle: oral, diet
Length of exposure (le) = 349 days
Length of experiment (Le) a 349 days
Ufespan of animal (L) = 730 days
Body weight = 0.35 kg (assumed)
Tumor site and type: mammary adenorcarcinoma
Experimental Dose
or Exposure
1000 ppm x 0.05
(food factor)
Transformed Dose
(mg/kg/day)
50
0
Incidence
No. Responding/No.
26/44
3/44
Tested
Unadjusted q-|* = 2.3270696x10'* (mg/kg/day)'1
Human q-|* = 1.2337681 (mg/kg/day)~l
0087d
-69-
03/31/88
-------
APPENDIX C
0
0
oo
0.
1
-J
o
1
1
o
CO
Summary Table for 3,3'-D1ch1orobenz1d1ne
Species
Inhalation Exposure
Subchronlc NA
Chronic NA
Carclnogenlclty ID
Oral Exposure
Subchronlc NA
Chronic NA
Carclnogenlclty rat
REPORTABLE QUANTITIES
Based on Chronic Toxlclty:
Based on Carclnogenlclty:
Exposure Effect
NA NA
NA NA
ID ID
NA NA
NA NA
1000 ppm nummary
In diet adenocarclnoroas
1000
10
RfD or qi* Reference
"
NA -: :^~' NA
NA NA
ID ID
NA NA
NA NA
1.2 (lug/kg/day)'1 Stula
et al.,
Stula
et al.,
Stula
et al..
1975
1978
1975
CO
oo
00
NA = Not applicable; ID = Insufficient data
------- |