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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 1n this document
and the dates searched are Included 1n "Appendix: Literature Searched."
Literature search material Is 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 1s sent to the Program Officer (OSWER).
Several quantitative estimates are presented provided sufficient data
are available. For systemic toxicants, these Include Reference doses (RfOs)
for chronic and subchronlc exposures for both the Inhalation and oral
exposures. The subchronlc or partial lifetime RfD Is 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 Hfespan. 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 1s the same as traditionally employed for chronic estimates,
except that subchronlc data are utilized when available.
In the case of suspected carcinogens, a carcinogenic potency factor, or
q-j* (U.S. EPA, 1980), Is 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. An RfD may also be derived for the noncardno-
genlc health effects of compounds that are also carcinogenic.
Reportable quantities (RQs) based on both chronic toxlclty and carclno-
genlclty are derived. The RQ Is used to determine the quantity of a hazard-
ous substance for which notification Is 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
IgnltabllHy, reactivity, aquatic toxlclty, and acute mammalian toxldty).
Chemlcal-spedf 1c 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.
111
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EXECUTIVE SUMMARY
4,4'-Methylene-b1s(2-chloroan1l1ne) 1s a solid at room temperature. It
Is slightly soluble In water and Is soluble in most common organic solvents
(Sax and Lewis, 1987; Wlndholz et al., 1983). According to U.S. EPA TSCA
production file, three companies produced or Imported between 2.01 and 20.1
million pounds of 4,4'-methylene-b1s(2-chloroan1l1ne) In 1977 (TSCAPP,
1989). U.S. production of this compound ceased by 1980. Since 1980, all
4,4'-methylene-b1s(2-chloroannine) used In the United States Is Imported
from Japan (Ward et al., 1987). 4,4'-Methylene-b1s(2-chloroannine) Is used
as a curing agent for both Hquld-castable polyurethane elastomers and epoxy
resins (Flshbeln, 1984; Sax and Lewis, 1987).
4,4'-Methylene-b1s(2-chloroan1l1ne) 1s expected to exist predominantly
In the partlculate form In the ambient atmosphere. Pertinent data regarding
the atmospheric fate of 4,4'-methylene-b1s(2-chloroan111ne) were limited In
the available literature. Deposition of both partlculate and adsorbed
4,4'-methylene-b1s(2-chloroan1l1ne) 1s expected to be the dominant fate
process 1n the atmosphere. The gas-phase reaction of 4,4'-methylene-b1s-
(2-chloroanlllne) with ozone Is not expected to be significant and the gas
phase reaction with photochemlcally produced H0» Is expected to be rapid
for the small proportion of this compound existing In the vapor phase. If
released to water, 4,4'-methylene-b1s(2-chloroan111ne) Is expected to adsorb
strongly to sediment and suspended organic matter. It may moderately
bloaccumulate In fish and aquatic organisms. Neither hydrolysis nor
volatilization to the atmosphere 1s expected to be significant. Conflicting
data on the blodegradatlon of 4,4'-methylene-b1s(2-chloroan1lIne) under
aerobic conditions were found; therefore, Its fate by this process 1s
1v
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unknown. 4,4'-Methylene-b1s(2-chloroan1l1ne) Is expected to adsorb strongly
to son. It may form covalent bonds with the active sites of the soil. A
single study suggests that aerobic blodegradatlon In soil may occur after a
short Induction period. 4,4'-Methylene-b1s(2-chloroan111ne) Is not expected
to volatilize from the soil surface to the atmosphere.
Limited data are available regarding exposure to 4,4'-methylene-b1s-
(2-chloroan1l1ne). It appears that occupational exposure by dermal contact
will predominate; however, Inhalation of partlculate 4,4'-methylene-b1s-
(2-chloroan1l1ne) Is also possible. High levels of this compound have been
found on Indoor surfaces where 1t Is stored or used commercially. The lack
of ambient monitoring data on levels of 4,4'-methylene-b1s(2-chloroanal1ne)
does not allow the determination of the level of human exposure. The
monitoring data located In the literature are concerned with the levels of
4,4'-methylene-b1s(2-chloroan1l1ne) near a major manufacturing site no
longer producing this compound. Thus, populations that reside near facili-
ties that use 4,4'-methylene-b1s(2-chloroan111ne) may be exposed to the
compound. Sufficient data could not be located to accurately predict levels
of exposure to the general population.
No data were located regarding the environmental toxldty of
4,4'-niethylene-b1s(2-chloroan1l1ne).
Excretion data on rats Indicate that 4,4'-methylene-b1s(2-chloroanH1ne)
Is rapidly absorbed from the gastrointestinal tract (Farmer et al.t 1981).
It has been shown that 4,4'-methylene-b1s(2-chloroan1l1ne) can be absorbed
through the skin of rabbits (E.I. DuPont de Nemours and Company, Inc.,
1977), dogs (Hanls et al., 1984) and humans (OsoMo et al., 1986). In the
dog study, absorption was estimated at 2.4-10% of the applied dose over a
24-hour exposure period. Absorbed 4,4'-methylene-b1s(2-chloroannine) 1s
-------
rapidly distributed throughout the body. Highest levels, regardless of
route of administration, are located In the liver and fat, but no organ or
tissue appears to preferentially accumulate or retain 4,4'-methylene-b1s-
(2-chloroan1lIne) or Its metabolites.
Farmer et al. (1981) determined that both low and high doses of 4.4'-
methylene-b1s(2-chloroanH1ne) were extensively metabolized by the same
metabolic systems 1n rats. They showed that the major urinary products were
conjugates of several metabolites and that only 1-2X was excreted unchanged
In the urine. In the urine of humans with known exposure to 4,4'-methylene-
b1s(2-chloroan1llne), however, only unmetabollzed compound was Identified In
the urine and the Investigators concluded that Important species differences
exist In the metabolism of the compound. Ijn vitro studies, however, Indi-
cate that liver mkrosomal preparations from rats and humans blotransform
4,4'-methylene-b1s(2-chloroannine) to the N-hydroxy, 6-hydroxy and benz-
hydrol derivatives (Morton et al., 1986).
Based on accidental human dermal exposure to 4,4'-methylene-b1s-
(2-chloroanHlne), the half-life In a human was estimated as -23 hours
(Osorlo et at., 1986). Studies using rats (Farmer et al., 1981; Tobes et
al., 1983) Indicate that excretion 1s rapid after Intravenous,
Intraperltoneal or oral administration. Fecal excretion exceeds urinary
excretion roughly by a factor of two regardless of route of administration.
i
A distribution study reported that 32% of the dose given to dogs was located
In the bile 24 hours after Intravenous treatment, Indicating that biliary
excretion Is Important In the elimination of 4,4'-methyelene-bls(2-chloro-
anlllne).
The carcinogenic effects of 4,4'-methylene-b1s(2-chloroan1llne) admin-
istered In the diets of mice, rats and dogs have been the subject of several
studies. Results have shown that administration 1n the diet to CD-I mice
v1
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produced increased Incidences of hepatomas In females compared with controls
(RussMeld et al., 1975) (see Table 6-1). When given 1n the diet to male
and female Charles River CD rats, 4,4'-methylene-b1s(2-chloroan1l1ne)
produced statistically significant Increased Incidences of lung adeno-
cardnomas compared with controls (Stula et al., 1975) (see Table 6-2).
Komm1nen1 et al. (1979) reported that dietary 4,4'-methylene-b1s(2-
chloroanlUne) given to male Charles River Sprague-Dawley rats for 18 months
followed by 6 months of observation led to Increased Incidences of pulmonary
adenomas and adenocarclnomas, mammary adenocardnomas, Zymbal gland carcino-
mas and hepatocellular carcinomas compared with controls (see Table 6-3).
Four of five female beagle dogs given 4,4'-methylene-b1s(2-chloro- aniline)
In gelatin capsules for <9 years developed urinary bladder tumors while no
control dogs developed such tumors (Stula et al., 1977) (see Table 6-4).
4,4'-Methylene-b1s(2-chloroan1l1ne) has been shown to be mutagenlc or
genotoxlc 1n bacterial and yeast assays, with or without mlcrosomal
activation (Takemura and Sh1m1zu, 1978; Ho et al., 1979; Shlmlzu et al.,
1982; McCann et al., 1975; Ichlnotsubo et al., 1981; Ho et al., 1979). It
failed to Induce sex-linked recessive lethal mutations In Drosophlla
melanogaster (Ho et al., 1979), and 1t did not produce chromatld aberrations
or SCE 1n Chinese hamster ovary cells or human leucocytes (Galloway et al.,
1985; Ho et al., 1979) (see Table 6-5). It was reported to produce
unscheduled DNA synthesis 1n rodent and rabbit hepatocytes (McQueen et al.,
1981, 1983; McQueen and Williams, 1982; MoM et al., 1988).
Occupational exposure to 4,4'-methylene-b1s(2-chloroan1l1ne) has been
associated with a reversible form of hematurla, but exposures were not
precisely quantified (Mastromatteo, 1965). Signs of systemic .toxlclty from
oral administration of 4,4'-methylene-b1s(2-chloroan111ne) In animals
-------
Included the following: liver Injury In female beagle dogs administered 7.3
mg/kg/day for 9 years (Stula et al., 1977); high mortality In female CD-I
mice at dietary concentrations of 2000 ppm for 18 months followed by 6
months on a normal diet (RussMeld et al., 1975); decreased body weight
gains In CD-I male rats administered 500 or 1000 ppm for 18 months followed
by 6 months of observation (Russfleld et al., 1975); decreased survival time
In CR Sprague-Dawley rats administered 4,4'-methylene-b1s(2-chloroan1l1ne)
In the diet at 0, 250, 500 and 1000 ppm for 18 months and observed for 6
months (Kommlnenl et al., 1979) and liver effects Including hepatocytome-
galy, fatty change, necrosis, bile duct proliferation and flbrosls In
Charles River CD rats administered 1000 ppm In the diets for <2 years (Stula
et al., 1975).
In vitro studies with rat liver cells (S1lk et al., 1989), and dog and
human bladder explant cultures (Stoner et al., 1987) have led to the
Isolation of 4,4'-methylene-b1s(2-chloroan1l1ne)-DNA adducts, suggesting
that 4,4'-methylene-b1s(2-chloroan1l1ne) can bind to DNA 1n these species.
Three of the adducts were common to dog and human cell cultures.
Because of positive evidence regarding the cardnogenlclty of
4,4'-methylene-b1s(2-chloroanH1ne) when administered orally to rats, mice,
and dogs, and when administered subcutaneously to rats, the compound was
assigned to U.S. EPA Group 82: probable human carcinogen. A human q,* of
l.SxlO"1 (mg/kg/day)"1 was derived for oral exposure to 4,4'-methylene-
b1s(2-chloroan1!1ne) and was also adopted for Inhalation exposure. An air
concentration of 5.4x10"" mg/m3 would be associated with Increased
cancer risk of IxlO"5 and a concentration of 2.7xlO~3 mg/8. In drinking
water would be associated with Increased cancer risk of IxlO"5. An RQ for
cardnogenlclty of 100 was based on the Incidence of lung tumors In rats In
studies by Stula et al. (1975) and Kommlnenl et al. (1979).
-------
Pertinent data regarding the developmental and reproductive toxlclty of
4,4'-methylene-b1s(2-chloroan1l1ne) were not located In the available
literature.
An RfD for subchronlc and chronic oral exposures of 0.0007 mg/kg/day was
based on the LOAEL that resulted 1n liver Injury and bladder Inflammation In
dogs treated for 9 years at 7.3 mg/kg/day (Stula et al., 1977). An RQ of
1000 for chronic (noncancer) toxldty was based on Increased mortality In
male rats given 4,4'-methylene-b1s(2-chloroan1l1ne) In the diet for 18
months (Kommlnenl et al., 1979).
1x
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TABLE OF CONTENTS (cont.)
Page
4. ENVIRONMENTAL TOXICOLOGY 11
4.1. AQUATIC TOXICOLOGY 11
4.1.1. Acute Toxic Effects on Fauna 11
4.1.2. Chronic Effects on Fauna 11
4.1.3. Effects on Flora 11
4.1.4. Effects on Bacteria 11
4.2. TERRESTRIAL TOXICOLOGY 11
4.2.1. Effects on Fauna 11
4.2.2. Effects on Flora 12
4.3. FIELD STUDIES 12
4.4. AQUATIC RISK ASSESSMENT. . 12
4.5. SUMMARY 12
5. PHARMACOKINETCS 13
5.1. ABSORPTION 13
5.2. DISTRIBUTION 14
5.3. METABOLISM 16
5.4. EXCRETION 19
5.5. SUMMARY 20
6. EFFECTS 22
6.1. SYSTEMIC TOXICITY 22
6.1.1. Inhalation Exposure 22
6.1.2. Oral Exposure 22
6.1.3. Other Relevant Information 25
6.2. CARCINOGENICITY 27
6.2.1. Inhalation 27
6.2.2. Oral 28
6.2.3. Other Relevant Information 35
6.3. MUTAGENICITY 37
6.4. DEVELOPMENTAL TOXICITY 41
6.5. REPRODUCTIVE TOXICITY 41
6.6. SUMMARY 41
7. EXISTING GUIDELINES AND STANDARDS 44
7.1. HUMAN 44
7.2. AQUATIC 44
x1
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TABLE OF CONTENTS (cent.)
Page
8. RISK ASSESSMENT 45
8.1. CARCINOGENICITY 45
8.1.1. Inhalation 45
8.1.2. Oral. 45
8.1.3. Other Routes 46
8.1.4. Weight of Evidence 46
8.1.5. Quantitative Risk Estimates 46
8.2. SYSTEMIC TOXICITY 49
8.2.1. Inhalation Exposure 49
8.2.2. Oral Exposure 49
9. REPORTABLE QUANTITIES 53
9.1. BASED ON SYSTEMIC TOXICITY 53
9.2. BASED ON CARCINOGENICITY 57
10. REFERENCES 59
APPENDIX A: LITERATURE SEARCHED 71
APPENDIX B: CANCER DATA SHEET FOR DERIVATION OF A q-|* USING
GLOBAL86 74
APPENDIX C: SUMMARY TABLE FOR MOCA 77
APPENDIX D: DOSE/DURATION RESPONSE GRAPH(S) FOR EXPOSURE TO
4,4'-METHYLENE-BIS(2-CHLOROANILINE) 78
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LIST OF TABLES
No. Title Page
5-1 Time-Dependent Tissue Distribution of Radioactivity In
Rats After Intravenous Administration of 0.49 mg/kg [14C]
Methylene-b1s(2-chloroan1l1ne) 15
5-2 Tissue Distribution of Radioactivity 1n Rats and Dogs After
Administration of [l4C]Methylene-b1s(2-Chloroan1l1ne) .... 17
6-1 Number of Tumors In Charles River CD-I Mice Fed Diets
Containing 4,4'-Methylene-b1s(2-Chloroan1l1ne) for 18
Months and Observed for an Additional 6 Months 29
6-2 Incidence of Tumors In Charles River CD Rats Fed 4,4'-
Methylene-b1s(2-Chloroan1l1ne) 1n the Diet for <2 Years ... 31
6-3 Incidence of Tumors In Male Charles River Sprague-Dawley
Rats Fed Diets Containing 4,4'-Methylene-b1s(2-chloroan1-
Une) for 18 Months then Observed for 5 Additional Months . . 33
6-4 Incidence of Tumors 1n Female Beagle Dogs Given Capsules
Containing 4,4'-Methylene-b1s(2-Chloroan1l1ne) for <9 Years . 36
6-5 Genotoxldty Testing of 4,4'-Methylene-b1s{2-Chloroan1l1ne) . 38
9-1 Oral Toxldty Summary for 4,4'-Methylene-b1s(2-Chloro-
anlUne) 54
9-2 Composite Scores for 4,4'-Methylene-b1s(2-Chloroan1l1ne). . . 55
9-3 4,4'-Methylene-b1s(2-Chloroan1l1ne): Minimum Effective
Dose (MED) and Reportable Quantity (RQ) 56
9-4 Derivation of Potency Factor (F) for 4,4'-Methylene-
b1s(2-ChloroanH1ne) 58
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LIST OF ABBREVIATIONS
AEL Adverse-effect level
BCF B1oconcentrat1on factor
bw Body weight
CAS Chemical Abstract Service .
CS Composite score
DMSO Dimethyl sulfoxlde
ONA Deoxyr1bonucle1c acid
PEL Frank effect level
GOT Glutamlc oxaloacetlc transamlnase
GP1 Glutamlc pyruvlc transamlnase
Koc Soil sorptlon coefficient standardized
with respect to organic carbon
Kow Octanol/water partition coefficient
1050 Dose lethal to 50% of recipients
LDH Lactate dehydrogenase
LED Lowest effective dose
LOAEL Lowest-observed-adverse-effect level
NOAEL No-observed-adverse-effect level
ppb Parts per billion
ppm Parts per million
MTD Maximum tolerated dose
RfD Reference dose
SCE Sister chromatld exchange
TLV Threshold limit value
TWA Time weighted average
UV Ultraviolet
xlv
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1. INTRODUCTION
1.1. STRUCTURE AND CAS NUMBER
4,4'-Methylene-b1s(2-chloroan1l1ne) Is known by the synonyms bis amlne
b1s(3-chloro-4-am1nophenyl)methane, b1s(4-am1no-3-chlorophenyl)methane and
methylene-b1s-ortho-chloroan1l1ne; the trade names Cuamlne MT, Curalln M,
Curene 442 and Cyanaset; and the acronyms CL-MDA and DACPM (Chemllne, 1989).
The structure, CAS number, empirical formula and molecular weight are given
below:
Cl Cl
CAS Registry number: 101-14-4
Empirical formula: ci3Hiocl2N2
Molecular weight: 267.16
1.2. PHYSICAL AND CHEMICAL PROPERTIES
4,4'-Methylene-b1s(2-chloroan1l1ne) 1s a solid at room temperature. It
1s slightly soluble In water and 1s soluble In dilute adds, ether, alcohol,
methylethylketone, acetone, esters and aromatic hydrocarbons (Sax and Lewis,
1987; Wlndholz et al., 1983). Selected physical and chemical properties of
4,4-methylene-b1s(2-chloroan1l1ne) are as follows:
Melting point:
Hater solubility
at 24°C:
Log Kow:
Vapor pressure at 25°C:
Conversion factors
at 25°C:
110°C
139 mg/l
3.94
1.32xlO"8 mm Hg
1 mg/m3 = 9.16x10~2 ppm;
1 ppm = 10.9 mg/m3
Wlndholz et al., 1983
Voorman and Penner,
1986
U.S. EPA, 1987a
U.S. EPA, 1987b
0237d
-1-
09/05/89
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1.3. PRODUCTION DATA
This compound was produced or Imported In 1977 by Polyester Corpora-
tion of Southampton, NY; Andersen Development Co., of Adrian, MI; and E.I.
Oupont and De Nemours and Co., of Deepwater, NJ. The total production
volume was between 2.01 and 20.1 million pounds (one plant's production
volume was listed as confidential) (TSCAAP, 1989). These companies ceased
production by 1980. Since 1980, all 4,4'-methylene-b1s(2-chloroan111ne)
used In the United States has been Imported from Japan (Ward et al., 1987).
It has been estimated that 200-400 U.S. firms are engaged In the production
of products cured with 4,4'-methylene-b1s(2-chloroan1l1ne) (Ward e-t al.,
1987).
4,4'-Methylene-b1s(2-chloroan1l1ne) was produced commercially In the
United States by the condensation of formaldehyde with two equivalents of
ortho-chloroanlUne (Flshbeln, 1984).
1.4. USE DATA
4,4'-Methylene-b1s(2-chloroan1l1ne) Is perhaps the most widely used
curing agent for both epoxy resins and Hquld-castable polyurethane elas-
tomers suitable for molded mechanical articles and for potting and encapsu-
lating purposes (Flshbeln, 1984; Sax and Lewis, 1987).
1.5. SUMMARY
4,4'-Methylene-b1s(2-chloroan1l1ne) Is a solid at room temperature. It
Is slightly soluble In water and 1s soluble 1n most common organic solvents
(Sax and Lewis, 1987; Wlndholz et al., 1983). According to U.S. EPA 1SCA
production file, three companies produced or Imported between 2.01 and 20.1
million pounds of 4,4'-methylene-b1s(2-chloroan1l1ne) 1n 1977 (1SCAPP,
1989). U.S. production of this compound ceased by 1980. Since 1980, all
4,4'-methylene-b1s(2-chloroan1l1ne) used In the United States Is Imported
0237d -2- 11/16/89
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from Japan (Ward et al., 1987). 4,4'-Methylene-b1s(2-chloroan1l1ne) Is used
as a curing agent for both I1qu1d-castable polyurethane elastomers and epoxy
resins (Flshbeln, 1984; Sax and Lewis, 1987).
0237d -3- 09/05/89
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2. ENVIRONMENTAL FATE AND TRANSPORT
2.1. AIR
The estimated vapor pressure of 4,4'-methylene-b1s(2-chloroan1l1ne),
1.32xlO"8 mm Hg at 25°C (U.S. EPA, 1987b), suggests that this compound
win exist partially 1n the vapor phase, but predominantly In the
partlculate form 1n the ambient atmosphere (Elsenrelch et a!., 1981).
2.1.1. Reaction with H0». Using a method of Atkinson (1985), the rate
for the gas phase reaction of 4,4' -methylene-b1s(2-chloroanH1ne) with
photochemically produced H0« can be estimated at 1.22xlO~10 cm3/
molecule-second. If the average atmospheric H0» concentration 1s 1x10*
molecules/cm3, then a half-life of 0.132 days can be calculated (Atkinson,
1985). However, 4,4'-methylene-b1s(2-chloroan1Hne) 1s expected to exist
predominately 1n the partlculate form In the ambient atmosphere and only
small amounts will exist 1n the vapor phase. Partlculate 4,4'-methylene-
b1s(2-chloroanH1ne) 1s expected to be less chemically reactive. The actual
rate of destruction of total 4,4'-methylene-b1s(2-chloroan1l1ne) by photo-
chemlcally produced HO- will therefore be considerably slower.
2.1.2. Reaction with Ozone. Quantitative data regarding the gas-phase
reaction of 4,4'-methylene-b1s(2-chloroan1l1ne) were not located In the
available literature cited In Appendix A. In water, 4,4-methylene-bls-
(2-chloroan1l1ne) 1n water has been reported to undergo <1% reaction with
ozone after 130 minutes (Fochtman and Elsenberg, 1979). Therefore, the gas
phase destruction of 4,4'-methylene-b1s(2-chloroan111ne) by the reaction
with ozone Is not expected to be a significant process.
2.1.3. Photolysis. Pertinent data regarding the direct destruction of
4,4'-methylene-b1s(2-chloroanH1ne) by photolysis were not located In the
available literature dted In Appendix A. Generally, aromatic amines adsorb
0237d -4- 11/16/89
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UV light at wavelengths >290 nm, which 1s the UV region of the electro-
magnetic spectrum found In the atmosphere. Therefore, 4,4'-methylene-b1s-
(2-chloroan1l1ne) may be a candidate for direct photochemical degradation 1n
the atmosphere.
2.1.4. Physical Removal Processes. Quantitative data regarding the
physical removal of 4,4'-methylene-b1s(2-chloroan1l1ne) from the atmosphere
were not located In the available literature cited In Appendix A. The water
solubility of 4,4'-methylene-b1s(2-chloroan1l1ne), 13.9 mg/l at 24°C
(Voorman and Penner, 1986), suggests that rain washout may occur. Dry
deposition of adsorbed partlculate 4,4'-methylene-b1s(2-chloroanH1ne) Is
also expected to be a significant removal process.
2.2. WATER
2.2.1. Hydrolysis. An experimental rate constant for the hydrolysis of
4,4'-methylene-b1s(2-chloroan1l1ne) at neutral pH has been determined as
<9xlO"8 I/hour, which 1s a half-life of >800 years. Under addle condi-
tions, an experimental rate of 2.9xlO~* l/mole-hour equals a half-life of
4000 years at pH 5 (Ellington et al., 1988). Thus, hydrolysis of
4,4'-methylene-b1s(2-chloroan1l1ne) 1s not expected to be a significant fate
process.
2.2.2. Oxidation. Pertinent data regarding the chemical oxidation of
4,4'-methyleneb1s(2-chloroan1l1ne) 1n water were not located In the
available literature cited In Appendix A.
2.2.3. Photolysis. Pertinent data regarding the photolytlc destruction
of 4,4'-methylene-b1s(2-chloroanH1ne) in water were not located In the
available literature cited 1n Appendix A.
2.2.4. H1crob1al Degradation. In a screening test using settled domestic
wastewater sludge under aerobic conditions, the author reported that
0237d -5- 11/16/89
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4,4'-methylene-b1s(2-chloroan1l1ne) was not biologically dlsslmllated
effectively after the sixth subculture and enrichment process (Tabak et a!.,
1980). In another static test using a settled domestic wastewater Inoculum,
2.0 mg/8. of 4,4'-methylene-b1s(2-chloroan11Ine) was not biologically
decomposed effectively under aerobic conditions. When the authors utilized
a continuous biological reactor with the same Inocula, 4,4'-methylene-b1s-
(2-chloroan1l1ne) was reduced from 2.02 to 0.09 mg/a, In 24 hours, which
the authors reported as loss from blodegradatlon (Fochtman and Elsenberg,
1979). However, no suitable control experiment was performed and It Is
possible that the loss of 4,4'-methylene-b1s(2-chloroan1l1ne) was due to
adsorption to the sludge rather than to blodegradatlon. Others have
reported that 4,4'-methylene-b1s(2-chloroan1l1ne) can concentrate In the
sludge of waste water treatment plants (Parr 1s et al., 1980).
Pure cultures of Norcardlopsls sp. and Bacillus megaterlum degraded
4,4'-methylene-b1s(2-chloroan111ne) to products resulting from N-acylat1on
or N-hydroxylat1on of the aromatic nitrogen (Yoneyama and Matsumura, 1984).
2.2.5. Bloconcentratlon. The BCF of an organic compound Is directly
related to Us K . Using the, regression equation log BCF = 0.76 log
KQW - 023 (Bysshe, 1982), a value of 581 can be calculated for
4,4'-methylene-b1s(2-chloroannine) using an estimated KQW of 3.94 (U.S.
EPA, 1987a). This value suggests that 4,4'-methylene-b1s(2-chloroan1lIne)
Is expected to moderately bloaccumulate In fish and aquatic organisms.
2.2.6. Adsorption. Since 4,4'-methylene-b1s(2-chloroan111ne) has a
strong affinity for soil (Section 2.3.2.), this compound Is expected to
significantly adsorb to sediment and suspended organic matter.
2.2.7. Volatilization. Using the bond contribution method of Mine and
Mookerjee (1975), a Henry's Law constant of 4.06X10"11 atm-mVmole can
0237d -6- 11/16/89
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be estimated for 4",4'-methylene-b1s(2-chloroan1l1ne). This value suggests
that volatilization from water to the atmosphere will not be an Important
process (Thomas, 1982); the volatilization rate will be so slow that
4,4'-methylene-b1s(2-chloroan1l1ne) will be expected to concentrate In
solution as the water evaporates.
2.3. SOIL
2.3.1. H1crob1al Degradation. 4,4'-Methylene-b1s(2-chloroan1l1ne) under-
went mkroblal degradation to b1s(2-chloroanH1ne)methone, the corresponding
benzophenone analog, In Hoytvllle soil under aerobic conditions after a
3-day Induction period. No rate Information was presented In this study and
complete mineralization of 4,4'-methylene-b1s(2-chloroanil1ne) to carbon
dioxide did not occur (Voorman and Penner, 1986).
2.3.2. Adsorption. 4,4'-Methylene-b1s(2-chloroan1l1ne) rapidly binds to
the soil matrix and probably forms covalent adducts with the active sites of
soil particles (Voorman and Penner, 1986). Generally, aromatic amines are
known to covalently bond to soil (Parrls, 1980). When 14C-labeled
4,4'-methylene-b1s(2-chloroan1l1ne) was applied to Hoytvllle soil at a
concentration of 4 mg/kg, only 30% of the original radioactivity was
extracted after 24 hours (extraction efficiency at time zero: 85-95%). At
an application rate of 40 mg/kg, -45% of the applied material was recovered
after 24 hours. The loss was not a result of blodegradatlon since the
Induction period 1n these experiments was 3 days (see Section 2.3.1.)
(Voorman and Penner, 1986). From the Freundllch constant (Kp=354) presented
In this paper, a K of 4600 can be calculated (Lyman, 1982). This KQC
value suggests that 4,4'-methylene-b1s(2-chloroan111ne) will display slight
mobility In soil (Swann et al., 1983).
0237d -7- 11/16/89
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2.3.3. Volatilization. The vapor pressure of 4,4'-methylene-b1s-
(2-chloroanlllne), 1.32xlO"8 mm Hg at 25°C (U.S. EPA, 1987b), In combina-
tion with Us strong adsorption to soil (Voorman and Penner, 1986), suggests
that volatilization from the soil surface to the atmosphere will not be
significant.
2.4. SUMMARY
4,4'-Methylene-b1s(2-chloroannine) Is expected to exist predominantly
1n the partlculate form 1n the ambient atmosphere. Pertinent data regarding
the atmospheric fate of 4,4'-methylene-b1s(2-chloroan1l1ne) were limited In
the available literature. Deposition of both partlculate and adsorbed-
4,4'-methylene-b1s(2-chloroanH1ne) Is expected to be the dominant fate
process In the atmosphere. The gas-phase reaction of 4,4'-methylene-b1s-
(2-chloroanlllne) with ozone 1s not expected to be significant and the gas
phase reaction with photochemlcally produced H0« 1s expected to be rapid
for the small proportion of this compound existing In the vapor phase. If
released to water, 4,4'-methylene-b1s(2-chlorban111ne) Is expected to adsorb
strongly to sediment and suspended organic matter. It may moderately
bloaccumulate In fish and aquatic organisms. Neither hydrolysis nor
volatilization to the atmosphere 1s expected to be significant. Conflicting
data on the blodegradatlon of 4,4'-methylene-bls(2-chloroan111ne) under
aerobic conditions were found; therefore. Us fate by this process Is
considered uncertain. In soil, 4,4'-methylene-b1s(2-chloroanll1ne) Is
expected to adsorb strongly to soil. It may form covalent bonds with the
active sites of soil particles. A single study suggests that aerobic
blodegradatlon 1n soil may occur after a short Induction period. 4,4'-
Methylene-b1s(2-chloroan1l1ne) Is not expected to volatilize from the soil
surface to the atmosphere.
0237d -8- 11/16/89
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3. EXPOSURE
3.1. WATER
At the site of a major producer of this compound near Adrian, MI,
4,4'-methylene-b1s(2-chloroanH1ne) was found 1n the Industrial lagoon
sediment at a minimum concentration of 1600 ppm (dry weight). It was
detected In the lagoon effluent water at a concentration of 250 ppb, and In
deep well water on the manufacturing site at a concentration of 1.5 ppb.
Surface runoff water at the plant contained 1 ppb of 4,4'-methylene-b1s-
(2-chloroan1l1ne). At the Adrian, MI, sewage treatment plant, the Influent
and effluent water had a detectable, but not quantifiable, amount of
4,4'-methylene-b1s(2-chloroanH1ne). The activated sludge from this treat-
ment plant had an estimated concentration of 18 ppm (dry weight) (Parrls et
al., 1980).
3.2. FOOD
Pertinent data regarding 4,4'-methylene-b1s(2-chloroan1l1ne) In fish
were not located In the available literature cited 1n Appendix A.
3.3. INHALATION
The lack of air monitoring data precludes the determination of worker
exposure to 4,4'-methylene-b1s(2-chloroan1l1ne) (Ward et al., 1987).
Occupational exposure by Inhalation of airborne dust can occur during the
transfer of 4,4'-methylene-b1s(2-chloroan1l1ne) from the container In which
1t was shipped (Schulte et al., 1988).
3.4. DERMAL
Although monitoring data are lacking, the most serious route of exposure
to 4,4'-methylene-b1s(2-chloroan111ne) Is believed to be through the skin
(Schulte et al., 1988). In a study attempting to correlate 4,4'-methylene-
b1s(2-chloroan1!1ne) air levels with Its concentration In the urine of
0237d -9- 09/05/89
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workers at a plant where H was used commercially, only 15% of the samples
were above the limits of analytical detection, 0.01 mg/m3. The concentra-
tion In the urine of workers, however, ranged from 70-1500 yg/l. This
was Interpreted as a result of absorption through the skin (Schulte et al.,
1988; Ward et al., 1987). Wipe samples of the work surfaces at facilities
using 4,4'-methylene-b1s(2-chloroan1l1ne) showed surface concentrations
<15,000 yg/100 cm2 In areas where 1t was stored or used (Ward et al.,
1987).
3.5. OTHER
The area surrounding an Adrian, MI, production facility of 4,4'-methyl-
ene-b1s(2-chloroanH1ne) had soil levels of this compound ranging from
1.6-200 ppm as monitored from a variety of samples (Flshbeln, 1984).
3.6. SUMMARY
Limited data are available regarding exposure to 4,4'-methylene-b1s-
(2-chloroan1l1ne). It appears that occupational exposure by dermal contact
will predominate; however, Inhalation of partlculate 4,4'-methylene-b1s-
(2-chloroan1l1ne) 1s also possible. High levels of this compound have been
found on Indoor surfaces where It Is stored or used commercially. The lack
of ambient monitoring data on levels of 4,4'-methylene-b1s(2-chloroanal1ne)
does not allow the determination of the level of human exposure. The
monitoring data located In the literature are concerned with the levels of
4,4'-methylene-b1s(2-chloroan1l1ne) near a major manufacturing site no
longer producing this compound. Thus, populations that reside near facili-
ties that use 4,4'-methylene-b1s(2-chloroan1l1ne) may be exposed to the
compound. Sufficient data could not be located to accurately predict levels
of exposure to the general population.
0237d -10- 09/05/89
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4. ENVIRONMENTAL TOXICOLOGY
4.1. AQUATIC TOXICOLOGY
4.1.1. Acute Toxic Effects on Fauna. Pertinent data regarding the
effects of acute exposure of aquatic fauna to 4,4'-methylene-b1s(2-chloro-
anlllne) were not located In the available literature dted 1n Appendix A.
4.1.2. Chronic Effects on Fauna.
4.1.2.1. TOXICITY ~ Pertinent data regarding the effects of chronic
exposure of aquatic fauna to 4,4'-methylene-b1s(2-chloroan1l1ne) were not
located In the available literature dted In Appendix A.
4.1.2.2. BIOACCUMULATION/BIOCONCENTRATION Pertinent data regarding
the bloaccumulatlon/bloconcentratlon potential of 4,4'-methylene-bls-
(2-chloroanlllne) In aquatic fauna were not located In the available
literature cited 1n Appendix A.
4.1.3. Effects on Flora.
4.1.3.1. TOXICITY Pertinent data regarding the toxic effects of
exposure of aquatic flora to 4,4l-methylene-b1s(2-chloroan1l1ne) were not
located 1n the available literature dted 1n Appendix A.
4.1.3.2. BIOCONCENTRATION « Pertinent data regarding the bloconcen-
tratlon potential of 4,4'-methylene-b1s(2-chloroan1l1ne) In aquatic flora
were not located In the available literature dted 1n Appendix A.
4.1.4. Effects on Bacteria. Pertinent data regarding the effects of
exposure of aquatic bacteria to 4,4'-methylene-b1s{2-chloroan1l1ne) were not
located In the available literature cited In Appendix A.
4.2. TERRESTRIAL TOXICOLOGY
4.2.1. Effects on Fauna. Pertinent data regarding the effects of
exposure of terrestrial fauna to 4,4'-methylene-b1s(2-chloroan111ne) were
not located 1n the available literature dted In Appendix A.
0237d -11- 09/05/89
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4.2.2. Effects on Flora. Pertinent data regarding the effects of
exposure of terrestrial flora to 4,4'-methylene-b1s(2-chloroan1l1ne) were
not located In the available literature cited In Appendix A.
4.3. FIELD STUDIES
Pertinent data regarding the effects of 4,4' -methylene-b1s(2-chloro-
anlllne) on flora and fauna 1n the field were not located In the available
literature dted 1n Appendix A.
4.4. AQUATIC RISK ASSESSMENT
No data were located regarding the effects of exposure of freshwater
fauna and flora to 4,4'-methylene-b1s(2-chloroan1l1ne). Acute studies with
representatives from eight families of freshwater fauna and at least three
chronic studies and one bloconcentratlon study with freshwater fauna and
flora are needed to develop a freshwater criterion by the method of U.S.
EPA/OWRS (1986).
Pertinent data regarding the effects of exposure of marine fauna and
flora to 4,4' -methylene-b1s(2-chloroanH1ne) were not located In the avail-
able literature cited In Appendix A. Acute studies with representatives
from eight families of marine fauna and at least three chronic studies and
/
one bloconcentratlon study with marine fauna and flora are needed to develop
a saltwater criterion by the method of U.S. EPA/OWRS (1986).
4.5. SUMMARY
No data were located regarding the environmental toxlclty of
4,4' -methylene-b1s(2-chloroanH1ne).
0237d -12- 09/05/89
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5. PHARMACOKINETICS
5.1. ABSORPTION
Farmer et al. (1981) administered to female LAC:Porton rats a single 10
mg/kg gavage dose of methylene-labeled [l4CJ4,4'-methylene-b1s(2-chloro-
anlllne) In arachls oil. Urine and feces were collected for 48 hours.
Urinary excretion accounted for 23.8% and fecal excretion for 59.5% of the
dose of radioactivity. Urinary excretion accounted for 21.IX of the dose at
24 hours. Radioactivity In several tissues accounted for -2.5% of the
dose... These Investigators also reported that fecal excretion accounted for
69.1% of the dose of radioactivity from a 1 mg/kg Intraperltoneal Injec-
tion. They concluded that gastrointestinal absorption was rapid.
Chin et al. (1983) studied the percutaneous absorption of [14C]methyl-
ene-labeled 4,4'-methylene-b1s(2-chloroan1l1ne) by exposing organ cultures
of human neonatal foreskin to [1*C]4,4l-methylene-b1s(2-chloroan1Hne)
bearing surfaces for <4 hours and monitoring the penetration of radio-
activity Into the cultures as a function of time and temperature. Foreskin
Integrity was Insured by hlstologlcal examination. Results showed that
[l4C]4,4'-methylene-b1s(2-chloroan1l1ne) was absorbed quickly and progres-
sively through the skin and that the rate of absorption was temperature
dependent. Thin-layer chromatography confirmed that 4,4'-methylene-
b1s(2-chloroanH1ne) was not metabolized by the foreskin organ cultures.
Accidental exposure data discussed In Section 5.4. (Osorlo et al., 1986)
provide qualitative evidence for percutaneous absorption In humans.
Following cutaneous application of -10.5 mg [14C]methylene-labeled
4,4'-methylene-b1s(2-chloroan1l1ne) in 0.5 ma. acetone to a 25 cm2 area
of shaved dog skin, radioactivity In whole blood or plasma was not detected
In the 24-hour monitoring period (Manls et al., 1984). At the end of 24
0237d -13- 11/16/89
-------
hours, urine contained 1.3% of the dose of radioactivity, bile contained
0.62% and skin at the application site contained 90%. By comparing
excretion following cutaneous and Intravenous administration, the authors
estimated that 2.4-10% of the administered dose was absorbed Into the
systemic circulation 1n 24 hours.
A study with rabbits showed that 4,4'-methylene-b1s(2-chloroan111ne) can
be absorbed dermally 1n this species as well (E.I. DuPont de Nemours and
Company, Inc., 1977). No other details of this study are available.
5.2. " DISTRIBUTION
A study measuring tissue distribution of [14C]4,4'-methylene-bls -
(2-chloroan1line) after a single Intravenous administration to rats was
conducted by Tobes et al. (1983). Twenty-one rats were lightly anesthetized
with ether and Injected with 0.49 mg/kg [14C]4,4'-methylene-b1s(2-chloro-
anHlne). After 10 minutes, 1-, 4- and 16-hour blood samples were taken,
and 5-6 rats/time period were anaesthetized with ether and sacrificed;
samples from selected tissues were excised, weighed and analyzed for total
radioactivity. Results are shown 1n Table 5-1. Because radioactivity was
seen In several tissues 10 minutes after administration, the authors
concluded that distribution occurred very quickly. Levels In adlposa and
skin higher at 1 hour than at 10 minutes suggested a shift in distribution
to organs of higher llpld content. Higher levels In the small Intestine
suggested enteric absorption of biliary excretion products. Analysis of
radioactivity levels In fractions of rat liver cells 1 hour after Intra-
venous administration Indicated that the activity was evenly distributed
within the cells.
Farmer et al. (1981) conducted a study to Investigate the distribution
of [J4C]4,4' -methylene-b1s(2-chloroan1Hne) In female LAC:Porton rats
0237d -14- 09/05/89
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TABLE 5-1
Time-Dependent Tissue Distribution of Radioactivity 1n Rats After
Intravenous Administration of 0.49 mg/kg [l4C]4,4'-Methylene-b1s-
(2-Chloroan111ne)
Tissue
Adipose
Adrenals
Bladder
Brain
grey
white
Intestine
large
small
Kidney
Liver
Lung
Muscle
Ovaries
Pancreas
Skin
Spleen
Stomach
Thyroid
Uterus
Blood
Mean
10 minutes
0.064
0.385
0.066
NR
NR
0.070
0.225
0.185
0.496
0.249
NR
0.194
0.204
0.080
0.087
0.158
NR
NR
0.090
Tissue Concentration, % kg
Time After Administration
1.0 hour 4
0.183
0.114
0.020
0.028
0.049
0.059
0.264
0.135
0.246
0.170
0.032
0.049
0.051
0.126
0.033
0.047
0.038
0.031
0.033
.0 hours
0.145
0.007
0.027
NR
NR
0.012
0.300
0.092
0.169
0.126
NR
0.040
NR
0.038
NR
0.041
NR
NR
0.027
dose/gb
16.0 hours
0.041
0.041
0.005
NR
NR
0.010
0.016
0.061
0.107
0.074
0.026
NR
0.006
NR
0.016
NR
NR
0.010
aSource: Tobes et al., 1983
bT1ssue concentrations are expressed as % kg dose/g to normalize differ
ences In animal weights.
NR = Not reported
0237d
-15-
11/16/89
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weighing 150 g. Two rats per dose group were given 1 mg/kg methylene-
labeled [l4C]4,4'-methylene-b1s(2-ch"loroan1l1ne) In arachls oil Intraperl-
toneal or 10 mg/kg orally. Manls et al. (1984) studied the distribution of
methylene-labeled [14C]4,4' -methylene-bls(2-chloroanH1ne) In groups of
four male, beagle-type mongrel dogs following Intravenous or cutaneous
administration of 10 mg 1n propylene glycol (Intravenous) or acetone (cuta-
neous). The results of both studies are shown 1n Table 5-2. For all routes
of administration, the liver and fat tissues showed the most radioactivity.
After 'Intravenous administration, the disappearance of radioactivity from
the blood was blphaslc, with a volume of distribution of 244 i for the
four dogs (Manls et al., 1984). Twenty-four hours after cutaneous adminis-
tration, amounts of radioactivity 1n the tissues were 10-20 times lower than
after Intravenous administration; 90% of the administered radioactivity was
located In the skin at the Injection site.
4,4'-Methylene-b1s(2-chloroan1l1ne) has been detected within human
erythrocytes (Williams, 1979). Because of their -120-day lifetime, they may
serve as storage depots for the compound.
5.3. METABOLISM
Farmer et al. (1981) (see Section 5.1.) partially Identified the metabo-
lites of [l4C]4,4'-methylene-b1s(2-chloroanH1ne) 1n the urine of rats
collected 5 days after treatment by IntraperHoneal Injection. Results
showed that 95-97% of the radioactivity In the urine of rats given 13 or 100
mg/kg [14C]4,4'-methylene-bls(2-chloroan1!1ne) was In the form of very
polar compounds, while 1-2% was tentatively Identified as 4,4'-methylene-
bls(2-chloroan1Hne). Two other less polar compounds were Isolated, but not
Identified. Deconjugatlon of the very polar metabolite fraction with a
sulphatase and glucuronldase mixture resulted In the Isolation of several
0237d -16- 09/05/89
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TABLE 5-2
Tissue Distribution of Radioactivity In Rats and Dogs After
Administration of [l4C]Methylene-b1s(2-Chloroan1lIne)
Tissue
Bile
Fat
Kidney
Liver
Lung
Sklnc
Blood
Feces
Urine
Total
Percentage
Rata
Oral
(10 mg/kg)
NR
0.22
0.16
1.91
0.07
NR
0.14
49.5
23.75
85.75
of Administered
1.p.
(1 mg/kg)
NR
0.43
0.26
2.27
0.18
NR
0.20_
69.1
29.45
101.89
Dose of Radloactlvl
Doqb
1.V.
(10 mg)
32
0.18
0.18
3.1
NR
NA
NR
NR
46
81.46
ty
p.c.
(10 mg)
0.62
NR
0.02
0.28
NR
90
NR
NR
1.3
92.22
aFarmer et al., 1981
bManis et al., 1984
cS1te of Injection
NR = Not reported; NA = not applicable; 1;p. = Intraperltoneal; 1.v. =.
Intravenous; p.c. = percutaneous
0237d -17- 09/05/89
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metabolites, one of which was Identified as 4,4'-methylene-b1s(2-chloro-
anlllne). Conjugated and unconjugated 4,4'-methylene-b1s(2-chloroan1l1ne)
constituted 3-6% of urinary radioactivity. Further analysis with glucuronl-
dase alone led the authors to conclude that the larger portion of the
conjugates were sulphates. Thus, It appeared that the low dose and high
dose of 4,4'-methylene-b1s(2-chloroan111ne) were extensively metabolized In
the rat by the same metabolic systems. The rapid urinary excretion of
radioactivity after treatment (Section 5.4.) suggested that metabolism was
rapid.. Farmer et al. (1981) also analyzed the urine of humans known to be
exposed to 4,4'-methylene-b1s(2-chloroan111ne). Unchanged parent compound
was Identified at levels <1500 nmol/J. (0.4 mg/l), but the metabolites
most prevalent 1n rat urine were not located 1n the human urine. The
Investigators concluded that Important species differences exist In the
metabolism of 4,4'-methylene-b1s(2-chloroan111ne) by rats and humans.
Manls and Braselton (1984) Identified the major 4,4'-methylene-b1s-
(2-chloroanlllne) metabolite In the urine of dogs In the Manls et al. (1984)
study as 5-hydroxy-3,3-d1chloro-4,4-d1am1nod1phenylmethane-5-sulfate, the
sulfate conjugate of a reactive Intermediate.
Morton et al. (1986) Investigated the formation of 4,4'-methylene-b1s-
(2-chloroan1l1ne) metabolites by Incubating rat or human liver mlcrosomes
with [l4C]4,4'-methylene-b1s(2-chloroan1l1ne) and appropriate cofactors
and then extracting and analyzing the products. Results showed that the
major product In both species was the n-hydroxy derivative, formed at rates
of 335 and -500 pmol/m1n/mg of protein In rats and humans, respectively.
Pretreatment of rats with phenobarbltal (but not with 3-methylcholanthrene)
resulted In a 4- to 8-fold Increase In the rate of formation of this
metabolite. This compound Induced unscheduled DNA synthesis In dog
0237d -18- 11/16/89
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urothellal cells. These findings support the possibility that
4,4'-methylene-b1s(2-chloroan1l1ne) Is metabolized by liver enzymes to a
compound that may be carclnocenlc to humans. Other shared metabolites
Included the benzhydol and the 6-hydroxy derivatives of 4,4'-methylene-bls-
(2-chloran1l1ne). The former was formed at rates of 82 and 60 pmol/mln/mg
of protein and the latter at 92 and 21 pmol/m1n/mg of protein In rats and
humans, respectively.
5.4. EXCRETION
When a worker 1n a 4,4'-methylene-b1s(2-chloroan1l1ne) production plant
was accidentally exposed to molten 4,4'-methylene-b1s(2-chloroanH1ne) on
his chest, abdomen, and extremities, urine levels of 4,4'-methylene-b1s-
(2-chloroan1l1ne) were monitored for 9 days and the half-life was determined
to be -23 hours, assuming a one-compartment model (Osorlo et al., 1986).
The amount of 4,4'-methylene-b1s(2-chloroanH1ne) In the urine of a man, who
was acddently exposed to the compound when hot liquid was sprayed over his
face and In his mouth, was measured for 20 days (Hoseln and Van Roosmalen,
1978). Data showed that the compound was excreted rapidly for the first 18
hours after exposure; urinary levels 2 weeks later were negligible.
Four female Sprague-Dawley rats were given 0.49 mg/kg [14CJ4,4'-
methylene-b1s(2-chloroan1!1ne) Intravenously. Twelve hours after dosing,
35.5% of the radioactivity had been excreted; by 24 hours, 79.4% had been
excreted, and by 48 hours, >90% of the administered radioactivity had been
excreted In the urine or feces. Fecal excretion predominated, accounting
for 73.4% of the dose at 48 hours (Tobes et al., 1983).
Farmer et al. (1981) (see Section 5.1.) measured the excretion of
radioactivity In feces and urine of rats for 48 hours after treatment with
[l4C]4,4'-methylene-b1s(2-chloroan1l1ne) at 1 mg/kg by Intraperltoneal
0237d -19- 11/16/89
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Injection or 10 mg/kg by gavage. Fecal excretion predominated, accounting
for 69.1% of the dose after IntraperUoneal administration and 59.5% after
oral treatment. Urinary excretion accounted for 29.5 and 23.8% after 1ntra-
perltoneal and oral treatment, respectively. Urinary excretion following
oral treatment was -90% complete at 24 hours.
Hanls et al. (1984) measured distribution of radioactivity 1n dogs 24
hours after Intravenous and percutaneous administration of [14C]4,4'-
methylene-b1s{2-chloroan1l1ne). Following Intravenous administration, 32%
of the administered dose was In the gallbladder bile and 46% was In the
urine, suggesting that biliary excretion may be an Important route of
elimination for 4,4'-methylene-b1s{2-chloroan1l1ne).
5.5. SUMMARY
Excretion data 1n rats Indicate that 4,4'-methylene-b1s(2-chloroan1l1ne)
1s rapidly absorbed from the gastrointestinal tract (Farmer et al., 1981).
It has been shown that 4,4'-methylene-b1s(2-chloroan1l1ne) can be absorbed
through the skin of rabbits (E.I. OuPont de Nemours and Company, Inc.,
1977), dogs (Manls et al., 1984) and humans (OsoMo et al., 1986). In the
dog study, absorption was estimated at 2.4-10% of the applied dose over a
24-hour exposure period. Absorbed 4,4'-methylene-b1s(2-chloroanH1ne) Is
rapidly distributed throughout the body. Highest levels, regardless of
route of administration, are located In the liver and fat, but no organ or
tissue appears to preferentially accumulate or retain 4,4'-methylene-bls-
(2-chloroanlllne) or Us metabolites.
Farmer et al. (1981) determined 1n rats that both low and high doses of
4,4'-methylene-b1s(2-chloroan1l1ne) were extensively metabolized by the same
metabolic systems. They showed that the major urinary products were conju-
gates of several metabolites and that only 1-2% was excreted unchanged 1n
0237d -20- 09/05/89
-------
the urine. In the urine of humans with known exposure to 4,4'-methylene-
b1s(2-chloroanH1ne), however, only unmetabollzed compound was Identified In
the urine and the Investigators concluded that Important species differences
exist In the metabolism of the compound. Iji vitro studies, however, Indi-
cate that liver mlcrosomal preparations from rats and humans blotransform
4,4'-methylene-b1s(2-chloroanH1ne) to the N-hydroxy, 6-hydroxy and benz-
hydrol derivatives (Morton et al., 1986).
Based on accidental human dermal exposure to 4,4'-methylene-b1s-
(2-chloroanlllne), the half-life In a human was estimated as ~23 hours
(Osorlo et at., 1986). Studies using rats (farmer et al., 1981; lobes et
al., 1983) Indicate that excretion 1s rapid after Intravenous, IntrapeM-
toneal or oral administration. Fecal excretion exceeds urinary excretion
roughly by a factor of two regardless of route of administration. A
distribution dog study reported that 32% of the dose was located In the bile
24 hours after Intravenous treatment. Indicating that biliary excretion Is
Important In the elimination of 4,4'-methylene-b1s(2-chloroan1l1ne).
0237d -21- 11/16/89
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6. EFFECTS
6.1. SYSTEMIC TOXICITY
6.1.1. Inhalation Exposure.
6.1.1.1. SUBCHRONIC Pertinent data regarding systemic toxldty
associated with subchronlc Inhalation exposure of humans or animals to
4,4'-methylene-b1s(2-chloroan1l1ne) were not located 1n the available
literature cited 1n Appendix A.
6.1.1.2. CHRONIC -- Llnch et al. (1971) conducted a cohort study of
62 workers at a 4,4'-methylene-b1s(2-chloroan1l1ne) manufacturing plant [31
were exposed to 4,4'-methylene-b1s(2-chloroan111ne) and 31 were not exposed]
to determine whether there was any evidence of chronic systemic disease In
the group. The length of 4,4'-methylene-b1s(2-chloroan1l1ne) exposure time
ranged from 6 months to 16 years. Although attempts were made to measure
concentrations of 4,4'-methylene-b1s(2-chloroanH1ne) vapor and dusts in the
air, the results were variable and unreliable. No differences were seen
regarding systemic Illnesses, urinary tract pathology, deaths or work
absenteeism. In another group of 178 workers who had worked with
4,4'-methylene-b1s{2-chloroan1l1ne) (but not for at least 10 years) there
were no differences In general health status compared with the entire plant
population. The authors concluded that no abnormal health effects In these
workers could be attributed to 4,4'-methylene-bls(2-chloroan11Ine) exposure.
In an earlier occupational study, Mastromatteo (1965) reported that
reversible hematurla had occurred In workers exposed to 4,4'-methylene-bls-
(2-chloroan1l1ne). Exposure levels, however, were not precisely quantified.
6.1.2. Oral Exposure.
6.1.2.1. SUBCHRONIC Pertinent data regarding systemic toxldty
associated with subchronlc oral exposure of humans or animals to
0237d -22- 09/05/89
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4,4'-methy1ene-b1s(2-chloroan111ne) were not located In the available
literature cited 1n Appendix A.
6.1.2.2. CHRONIC -- Stula et al. (1977) administered 4,4'-methylene-
b1s(2-chloroan1l Ine) (purity 90%) In gelatin capsules at 100 mg/day to a
group of six female purebred beagle dogs, 3 days/week for 6 weeks and then 5
days/week for <9 years. The average dally dose, calculated from data
provided by the Investigators, was 7.3 mg/kg. Another group of six dogs
that did not receive 4,4'-methylene-b1s(2-chloroan1l1ne) served as negative
controls. Dogs were weighed weekly and clinical blood chemistry and urine
parameters were analyzed regularly. After 9 years of treatment, surviving
dogs were sacrificed and necropsled. Major tissues were examined for gross
and microscopic lesions. There were no treatment-related effects on
mortality or body weights. Toxic signs noted In treated dogs Included
folUcular cystitis, Increased GPT (which usually Indicates liver Injury)
and liver nodular hyperplasla.
Russfleld et al. (1975) administered 4,4'-methylene-b1s(2-chloroan1l1ne)
In the diets to groups of Charles River CD-I mice (25 males and 25 females/
group) and male Charles River CD-I rats (25/group) for 18 months; the mice
and rats were observed for an additional 6 months. Treated animals were fed
a commercial diet supplemented with concentrations of 500 and 1000 ppm
(rats) or 1000 and 2000 ppm (mice). Control animals received the commercial
diet without the 4,4'-methylene-b1s(2-chloroan1l1ne). After 18 months of
treatment, all animals were given commercial diets without 4,4'-methylene-
b1s(2-chloroan111ne) for 6 months. Surviving animals were sacrificed,
necropsled, and major organs were subjected to hlstopathologlcal examina-
tion. Animals were weighed at regular Intervals during the treatment and
food consumption was monitored. Data for these parameters were not
0237d -23- 11/16/89
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reported. Results showed that 4,4'-methylene-b1s(2-chloroan1l1ne) treatment
had no effect on the survival of the rats. At the end of 18 months of
treatment, 96% of the controls and 80% of the treated rats survived. At the
end of 20-22 months, 55% of each group survived. At the end of the
treatment, body weights of the rats 1n the low-dose group averaged -50 g
lower than those of controls, while body weights of the high-dose group
averaged 100 g lower than those of controls; these differences persisted
until the end of the study. There were no "striking differences" among
treated groups of rats and controls In the Incidence of noncancer gross and
hlstopathologlcal lesions. Survival of female mice receiving the high
concentration of 4,4'-methylene-b1s(2-chloroan1l1ne) appeared affected by
treatment, since 14/25 mice 1n this group survived the study compared with
21/25 of the low-dose group and 20/25 controls. Of the male mice In the
study, 18/25 controls, 13/25 In the low-dose group and 20/25 In the
high-dose group survived. No treatment-related effects on body weight gain
In the mice were evident. Treated mice exhibited a lower Incidence and
Intensity of amyloldosls than controls.
Komm1nen1 et al. (1979) assessed the effect of 4,4'-methylene-b1s-
(2-chloroan1l1ne) administered In the diets to Charles River Sprague-Oawley
male rats on survival, body weight gain and hemoglobin and hematocrlt
parameters. Groups of 50-100 rats were given semi-purified nutritionally
adequate diets containing 0, 250, 500 or 1000 ppm of 4,4'-methylene-b1s-
(2-chloroanlllne) for 18 months, after which they received the basal diet
without 4,4'-methylene-b1s(2-chloroan1l1ne) for an additional 6 months.
Food consumption, body weights, hematocrlt and hemoglobin measurements were
made periodically, either on Individual rats or on 10 rats/group. Urine
samples from 10 rats were collected periodically and urine was analyzed for
volume and specific gravity. All rats that died before the conclusion of
0237d -24- 11/16/89
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the study were autopsled. All that survived until the end of the study were
sacrificed and autopsled, and gross lesions and major organs were examined
microscopically. Results showed that survival time decreased as dietary
amounts of 4,4' -methylene-b1s(2-chloroan111ne) Increased. Mean survival
times were 88.9, 86.6, 80.4 and 65.3 weeks for the controls to highest dose
groups, respectively. Differences In survival between controls and the 500
ppm dose group, and controls and the 1000 ppm dose group were statistically
significant (p<0.01 and p<0.001, respectively). The mean body weight gain.
of rats In the 1000 ppm group was lower than that of rats 1n the other
groups after 8 weeks of 4,4'-methylene-b1s(2-chloroan1l1ne) administration
until the end of the study. Food consumption was usually <11% different
from that of control rats. Body weight gain of rats 1n the other groups was
similar to that of the control rats throughout the study. HematocrH and
hemoglobin values were slightly less 1n the 1000 ppm group than In controls,
but the values were within normal ranges observed In this laboratory.
Noncancer results of pathological examinations were not reported.
A study by Stula et al. (1975), In which 4,4'-methylene-b1s(2-chloro-
anlUne) was given to groups of 50 male and 50 female Charles River CO rats
at dietary concentrations of 0 or 1000 ppm for <2 years, reported liver
changes Including hepatocytomegaly, fatty change, necrosis, bile duct
proliferation and flbrosls 1n rats that received 4,4'-methylene-b1s-
(2-chloroan1l1ne). Although statistical analysis was not performed, days on
test to 50% survival and the average number of days on test suggest that
survival was reduced In treated rats of both sexes compared with controls.
6.1.3. Other Relevant Information. A worker In an 4,4'-methylene-bls-
(2-chloroan1l1ne) production plant was exposed to 4,4'-methylene-bls-
(2-chloroanlHne) when he was accidentally sprayed with molten
4,4'-methy1ene-b1s(2-chloroan1l1ne) while cleaning a clogged delivery line.
0237d -25- 11/16/89
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Exposure to the chest, abdomen and extremities occurred over several
seconds. According to the report, no 4,4'-methylene-b1s(2-chloroan111ne)
was Ingested. After the 4,4'-methylene-b1s(2-chloroan1l1ne) had been gently
washed from the skin, the worker was described as having slight erythema and
a burning sensation. Laboratory results revealed normal renal and liver
function tests, no methemogloblnemla and no protein or red blood cells 1n
the urine. Urinary levels of 4,4'-methylene-b1s(2-chloroan1l1ne) were 1707
ppb 9 hours after exposure, but were nondetectable after 11 days. In this
worker, no acute effects from the 4,4'-methylene-b1s(2-chloroan1l1ne)
exposure were observed (Osorlo et al., 1986).
According to L1nch et al. (1971), human exposure to 4,4'-methylene-b1s-
(2-chloroanlllne) In sufficient amounts may cause cyanogenlc effects.
Employees In 4,4'-methylene-b1s(2-chloroan11Ine) production plants have had
urinary levels of 4,4'-methylene-b1s(2-chloroan1l1ne) as high as 25 mg/a.
without other observable symptoms of exposure (Linen et al., 1971).
One study reported the oral LD5Q In male rats as 750 mg/kg (Miller and
Sherman, 1965), but no other details of this study are available. Another
report gave the approximate lethal dose for rats as 1000 mg/kg; signs of
toxlclty Included polyuMa, cyanosis, weakness and pallor. Doses of 200
mg/kg every day for 10 days resulted In cyanosis, pallor, growth depression
and blood and urine abnormalities, but not mortality (Relnke, 1963).
Salamone (1981) reported an IntraperHoneal LD5Q of 64 mg/kg for
4,4'methylene-b1s(2-chloroan1l1ne) 1n DMSO 1n mice (gender not specified)
observed for <7 days after treatment.
S1lk et al. (1989) conducted a study to measure the binding of
4,4'-methylene-b1s(2-chloroanH1ne) to rat liver DNA Ui vivo and binding of
0237d -26- 11/16/89
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the N-hydroxylated derivative jji vitro. For the j_n vivo experiments, male
Wlstar rats were given an 1ntraper1toneal Injection of radlolabeled compound
1n DMSO. After 24 hours, the rats were sacrificed and the livers removed
for DNA extraction, Isolation and purification. For j_n vitro experiments,
rat liver slices were Incubated for 2.5 hours at 37°C with radlolabeled
compound. The DNA was then extracted and purified. Results showed that
three DNA adducts formed when 4,4'-methylene-b1s(2-chloroan1l1ne) was
Injected Intraperltoneal Into rats or when 1t was Incubated with rat liver
slices. One of the adducts seemed to result from n-hydroxylatlon of
4,4'-methylene-b1s(2-chloroan1l1ne); the other two were not Identified. In
a study designed to compare DNA binding and DNA-adduct formation of
4,4'-methylene-b1s(2-chloroan1l1ne) 1n explant cultures of human and dog
bladder, Stoner et al. (1987) found that 4,4'-methylene-b1s(2-chloroan1l1ne)
does bind to DNA of both species. The amount of binding was related to the
concentration of 4f4'-methylene-b1s(2-chloroannine) used. Of several
4,4'-methylene-b1s(2-chloroannine)-DNA adducts Identified, three were found
to be formed 1n both species.
6.2. CARCINOGENICITY
6.2.1. Inhalation. Linen et al. (1971) conducted a cohort study of 62
workers (see Section 6.1.1.2.) at a 4,4'-methylene-b1s(2-chloroan1l1ne)
manufacturing plant [31 were exposed to 4,4l-methylene-b1s(2-chloroan1l1ne)
and 31 were not exposed] to determine whether there was any human carcino-
genic potential. The length of 4,4'-methylene-b1s(2-chloroan1l1ne) exposure
time ranged from 6 months to 16 years. No difference In the Incidence of
malignant tumors In the two groups was observed.
NIOSH (1986, 1987) diagnosed two cases of bladder cancer (the
theoretical "expected number" of bladder cancer cases Is 0.39) Involving
nonsmoking men under 30 years of age among a cohort of 370 workers exposed
0237d -27- 07/23/90
-------
to 4,4'-methylene-b1s(2-chloroan1l1ne) who submitted to a urine cytology
screening examination. The screening test revealed neither positive nor
suspicious cases. The first case was Identified by the examination of an
Individual who was diagnosed with low-grade Intermittent hematuMa. The
diagnosis of this case provided the Impetus to offer cystoscoplc examination
to workers with atypical cells or hematurla on the cytology examination and
to an equal number of workers who had highest exposures to 4,4'-methylene-
b1s(2-chloroan1llne). The second case was diagnosed during the cystoscoplc
examination of 41 workers. NIOSH (1987) expressed concern because
4,4'-methylene-b1s(2-chloroan1l1ne) 1s structurally similar to benzldlne,
which Is known to cause bladder cancers In humans, and because animals
exposed to 4,4'-methylene-bls(2-chloroan1l1ne) developed bladder tumors.
6.2.2. Oral. Russfleld et al. (1975) (see Section 6.1.2.2.) gave
4,4'-methylene-b1s(2-chloroan1l1ne) In the diets to groups of 25 male and 25
female Charles River CD-I mice and 25 Charles River CD-I male rats.
Animals, 4-6 weeks old at the beginning of the study, were fed a commercial
diet for 2 weeks, then were fed the commercial diet supplemented with
concentrations of 500 or 1000 ppm (rats) or 1000 or 2000 ppm (mice)
4,4'-methylene-bls(2-chloroan1l1ne) for 18 months. Control animals received
the commercial diet without 4,4' -methylene-bls(2-chloroanH1ne). After 18
months of treatment, all animals were given commercial diets without
4,4'-methylene-b1s(2-chloroan1l1ne) for 6 months. Surviving animals were
then sacrificed, necropsled and major organs examined hlstopathologlcally.
Animals were scored for tumor formation at the end of the study. Of the
female mice, 0/20 controls, 9/21 at the low dose and 7/14 at the high dose
had hepatomas. Results are shown In Table 6-1. The differences between the
numbers of hepatomas seen In the treated females compared with the numbers
0237d -28- 11/16/89
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TABLE 6-1
Number of tumors In Charles River CD-I Mice Fed Diets Containing
4,4-Methylene-b1s{2-chloroan1llne) for 18 Months and Observed
for an Additional 6 Months3
Sex
F
F
F
M
M
Concentration
(ppm)
0
1000
2000
0
1000
2000
0
1000
2000
0
1000
2000
0
1000
2000
Tumor Type
hepatoma
hemangloma
hemanglosarcoma
hemangloma
hemanglosarcoma
Incidence
of Tumors
0/20
9/21b
7/14b
0/20
0/21
4/14
0/20
0/21
2/14
0/20
2/21
5/14
0/20
1/21
3/14
QUALITY OF EVIDENCE
Strength of study: The compound was administered to both sexes at two dose
levels and was 97% pure. Natural route of exposure;
adequate duration of exposure
Weakness of study: Small number of animals per dose group; MID may have
excluded
Overall adequacy: Adequate
\
aSource: RussHeld et al., 1975
bSlgn1f1cantly different from control (p<0.01)
0237d -29- 09/05/89
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seen In the controls were statistically significant (p<0.01). There also
appeared to be an Increased Incidence of hemanglomas and hemanglosarcomas In
treated mice, compared with controls. While the numbers of vascular tumors
observed l.n male and female treated mice were greater than those In the
controls, they were not greater than those observed In historical controls
and were not considered significant by the authors. Mice that received
4,4'-methylene-b1s(2-chloroan1l1ne) In their diets had smaller Incidences
of amyloid tumors than did controls (14/38 controls, 7/34 low dose, 4/34
high dose); no explanation for this decrease was given (Russfleld et al,
1975). Of the rats that survived, 0/22 controls, 1/22 at the low dose, and
4/19 at the high dose had hepatomas; 0/22 controls, 3/22 at the low dose and
4/19 at the high dose had adenomatosls of the lung. The authors Indicated
that the difference between the Incidence of tumors 1n the treated groups
and that of controls was not statistically significant. Other neoplasms
seen In treated rats but not the controls Included a tumor of the ear duct
and adenocardnomas of the lung, stomach, small Intestine and urinary
bladder.
Stula et al. (1975) studied the effects of 4,4'-methylene-b1s(2-chloro-
/
aniline) (-95% pure) on tumor formation In groups of 50 male and 50 female
Charles River CD rats when given at concentrations of 0 or 1000 ppm In the
diet for <2 years. After 1 year of treatment, 6 rats/group were sacrificed
and necropsled. All rats were necropsled, either at time of death during
the study or at terminal sacrifice, and 30 organs were sampled for hlsto-
loglcal examination. Results (Table 6-2) showed that treated rats had a
higher Incidence of lung adenocardnoma than controls. Adenomatosls was
observed after 1 year of treatment. In rare cases, a pleural blphaslc tumor
accompanied the lung tumors. The Incidence of the lung adenocardnomas was
statistically significant (p<0.05) compared with controls In both sexes.
0237d -30- 11/16/89
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TABLE 6-2
Incidence of Tumors 1n Charles River CD Rats Fed 4,4-Methylene-bls-
(2-Chloroan1l1ne) 1n the Diet for <2 Years*
Sex '
F
F
M
M
Concentration
(ppm)
0
1000
0
1000
Tumor Type Incidence
of Tumors
lung adenocardrioma 0/44
21/44
0/44
27/44
p Value
NA
p<0.05
NA
p<0.05
QUALITY OF EVIDENCE
Strengths of Study: Chemical 95% pure, adequate number of animals per
group; natural route of exposure, adequate duration of
exposure, MTD reached
Weakness of Study: Compound administered at only one dose level .
Overall Adequacy: Adequate
*Source: Stula et al., 1975
NA = Not applicable
0237d -31- 11/16/89
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Liver tumors were also observed In treated rats of both sexes at Incidences
greater than In controls; however, the differences were not statistically
significant. The Incidence of other types of tumors In the treated rats was
not different from the Incidence 1n control rats.
A study by Kommlnenl et al. (1979) assessed the effect of 4,4'-methyl-
ene-b1s(2-chloroanH1ne) given orally on tumor formation In Charles River
Sprague-Oawley male rats. The rats, 5 weeks old, were divided Into four
treatment groups consisting of 50-100 animals. Each group received
different dietary amounts of 4,4'-methylene-b1s(2-chloroan1!1ne). Dietary
concentrations of 0, 250, 500 or 1000 ppm of 4,4' -methylene-b1s(2-chloro-
anlUne) were given to the rats for 18 months. After this time, the regular
diet without 4,4'-methylene-b1s(2-chloroan1l1ne) was fed for an additional 6
months. Every 4 weeks, Individual rats were scored for size and location of
palpable masses. All rats that died before the conclusion of the study were
autopsled. All that survived until the end of the study were sacrificed and
autopsled. Gross lesions and major organs were examined microscopically.
Results of the carclnogenlcHy study are shown 1n Table 6-3. Increased
Incidences of pulmonary adenomas and adenocardnomas, mammary adenocard-
nomas, Zymbal gland carcinomas and hepatocellular carcinomas were seen In a
dose-related fashion and were attributed to administration of 4,4'-methyl -
ene-b1s(2-chloroanH1ne). Metastasis of these neoplasms to other organs,
such as kidneys, pituitary gland and pancreas, was also noted.
Stula et al. (1977) administered 4,4'-methyelne-b1s(2-chloroanll1ne)
(90% pure) at 100 mg/day In gelatin capsules to a group of six female
purebred beagle dogs on 3 days/week for 6 weeks and then on 5 days/week for
<9 years (average dally dose, 7.3 mg/kg). Another group of six dogs that
did not receive 4,4'-methylene-b1s(2-chloroan1l1ne) served as controls.
0237d -32- 09/05/89
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o
CO
j
Q.
TABLE 6-3
Incidence of Tumors In Male Charles River Sprague-Dawley Rats Fed Diets Containing
4,4-Methylene-b1s-(2-Chloroan1l1ne) for 18 Months then Observed for 5 Additional Months3
I
CO
CO
I
er>
CD
CO
Concentration Tumor Type
(ppm)
0 lung adenocar1nomasd
250
500
1000
0 all primary lung
250 neoplasms
500
1000
0 mammary adenocarclnomas
250
500
1000
0 Zymbal gland carcinomas
250
500
1000
0 hepatocel lular carcinomas
250
500
1000
Incidence**
(X)
0
14
17
62
1
23
37
70
1
5
11
28
1
8
7
22
0
3
4
36
Incidence0
(number bearing tumors/
number necropsled)
0/100
14/100
20/75
31/50
1/100
23/100
28/75
35/50
1/100
5/100
8/75
14/50
1/100
8/100
5/75
11/50
0/100
3/100
3/75
18/50
p Value
(compared with control)
NA
p<0.001
p<0.001
p<0.001
NA
p<0.001
p<0.001
p<0.001
NA
NS
p<0.01 and >0.001
p<0.001
NA
p<0.05 and >0.01
NS
p<0.001
NA
NS
NS
p<0.001
-------
o
ro
CO
TABLE 6-3 (cont.)
QUALITY OF EVIDENCE
The compound was administered at three dose levels. Adequate numbers of animals per
group; natural route of exposure; adequate duration of exposure, adequate survival,
MTD reached
Compound administered to male rats only; purity of compound not reported
Adequate
Strengths of study:
Weakness of study:
Overall adequacy:
aSource: Kommlnenl et al., 1979
bOata provided by Investigators
wcObta1ned by multiplying Incidence data expressed as percent provided by Investigators by the number
i necropsled and rounding to nearest Integer
^Includes bronchlolar-alveolar cell carcinomas
NA = Not applicable; NS = not stated
o
IT)
o
in
CO
-------
After 8.3 or 9 years of treatment, five surviving dogs of the treated group
and all control dogs were killed and necropsled. Major tissues were
subjected to hlstopathologlcal examination. Results are shown In Table
6-4. After 8.3-9 years of treatment, four of five treated dogs, but no
controls, had carcinomas of the urinary bladder.
6.2.3. Other Relevant Information. A group of 25 male and 25 female
Wlstar rats was given a low protein diet containing 0.1% 4,4'-methylene-b1s-
(2-chloroanHlne) for 500 days; a control group of rats was given a similar
diet 'without 4,4'-methylene-b1s(2-chloroannine) (Grundmann and Stelnhoff,
1970; Stelnhoff and Grundmann, 1971). After 500 days, all rats received the
low-protein diet without 4,4'-methylene-b1s(2-chloroan1l1ne) for the
remainder of their lives. The cumulative dose 1n the treated group was
estimated at 27 g/kg. Among treated rats, 23 males and 20 females died with
tumors; 22 and 18, respectively, were liver tumors. Thirteen treated rats
(8 males, 5 females) had primary lung tumors and 10 of these also had
hepatomas. Two mammary adenomas were additionally reported In the group.
The average survival times were 730, 565 and 535 days for controls, treated
males and females, respectively.
When 94% pure 4,4'-methylene-b1s(2-chloroan1l1ne) as a suspension
(vehicle not reported) was given to 17 male and 17 female Wlstar rats by
weekly subcutaneous Injections of 500 or 1000 mg/kg (total dose of 25 g/kg
bw), 9 rats developed liver cell carcinomas and 7 developed primary lung
carcinomas; 22 rats died with a total of 29 malignant tumors. Thirteen
malignant tumors, Including 1 lung tumor, developed 1n 50 control rats that
survived <1040 days. No malignant tumors of the liver were seen (Stelnhoff
and Grundmann, 1971).
0237d -35- 11/16/89
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TABLE 6-4
Incidence of Tumors 1n Female Beagle Dogs Given Capsules Containing
4,4-Methylene-b1s(2-chloroan1l1ne) for <9 Years3
Dose
(mg/kg/day)
0
7.3b
Tumor Type
urinary bladder carcinoma
Tumor Incidence
0/6
4/5
Strengths of Study:
Weakness of Study:
QUALITY OF EVIDENCE
Chemical was 90% pure; natural route of exposure;
adequate duration of exposure
Compound administered at one dose level; only female
animals used; small number of animals
Overall Adequacy: Inadequate
aSource: Stula et al., 1977
bl!me weighted, expanded average dose for dogs given 100 mg capsule 3
days/week for 6 weeks, then 5 days/week for 9 years (calculated from data
provided by Investigators)
0237d
-36-
09/05/89
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6.3. MUTAGENICITY
Results of mutagenldty testing of 4,4'-methylene-b1s(2-chloroan111ne)
are shown 1n Table 6-5. It has been reported to be positive In the
following bacterial and fungi tests: (1) In reverse mutation assays with
Salmonella typhlmuMum strains TA98 and TA100, with and without mlcrosomal
activation (Takemura and Sh1m1zu, 1978; Ho et al., 1979; Sh1m1zu et al.f
1982; McCann et al., 1975; Brooks and Dean, 1981); (2) 1n a reverse mutation
test with Escher1ch1a coll (Matsushlma et al., 1981); (3) In a phage
Induction test with E.. coll (Thomson, 1981); (4) 1n an E.. coll DNA repair
assay with mlcrosomal activation (Ichlnotsubo et al., 1981); (5) In a
rec-assay with Bacillus subtms (Kada, 1981); (6) In m1tot1c gene
conversion and mltotlc aneuploldy tests with Saccharomyces cerevlslae (Sharp
and Parry, 1981; Parry and Sharp, 1981); and (7) 1n a yeast mutation assay
with and without activation (Ho et al., 1979). It has given mixed responses
In assays with cells from more complex organisms. It was positive In cell
transformation assays with Syrian hamster embryo and Syrian hamster kidney
cells (Casto, 1980; Purchase et al., 1978} and Balb/3T3 cells (Dunkel et
al., 1981). It failed to Induce sex-linked recessive lethal mutations In
Drosophlla melanogaster (Ho et al., 1979). It did not produce chromosomal
aberrations or sister chromatld exchanges (SCE) In Chinese hamster ovary
cells or human leucocytes (Galloway et al., 1985; Ho et al., 1979). It was
reported to produce unscheduled DNA synthesis (DOS) 1n rodent and rabbit
hepatocytes (McQueen et al., 1981, 1983; McQueen and Williams, 1982; Mori et
al., 1988).
In comparative studies with hamster embryo cells and human male embryo
lung cells (Casto, 1983), hamster cells were more sensitive to the lethal
effects of 4,4'-methylene-b1s(2-chloroan1l1ne), the ID values
(yg/mi) being 45 and 270, respectively, after 2 hours of treatment.
0237d -37- 07/23/90
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TABIE 65
Genotoxlclty Testing of 4,4'-Hethy1ene-b1s(2-ch1oroan111ne)
CO
OL.
t
a
00
i
o
to
o
in
\
CD
U3
Assay
Reverse
nutation
Reverse
nutation
Reverse
nutation
Reverse
nutation
Reverse
nutation
ONA repair
Reverse
mutation
Prophage
Induction
Rec -assay for
DNA damage
Indicator /Organism
Salmonella
typhlmurlum
TA98
TA100
S. typhlmurlun
^strain not
specified)
S. typhlmurlum
TA1535
TA1537
TA1538
TA98
TA100
TA92
S. typhlmurlun
TA98
TA100
S. typhlmurlun
TA100
Escherlchla coll
E. coll
WP28/r
HP2uvrA
WP2uvrA/pKH101
E. coll
Lambda-sensitive
Bacillus subtil Is
HHrec* and
M«5rec-
Appllcatlon
NR
NR
prelncubatlon
plate
Incorporation
NR
plate
Incorporation
spot test
prelncubatlon
plate
Incorporation
liquid
suspension
filter disc
test
Concentration Activating Response
or Dose System
NR S-9
-
*
NR mlcrosomal »
0.2-2000 vg/plate
7s- -
7s-
»S »
7s- *
»S -
10-1000 yg/plate »S-9
*
»S-9
NR none »
NR S-9 »
NR
»S-9 -/NT
»S-9
7S -9 *
1000 or 2000 yg/mt iS-9 »
1 mg/dlsc »S-9 »
.5-9
Comment
NC
NC
Vehicle probably OHSO;
lowest effective dose
not reported
0.53 reversions/)!?,
2.74 reverslons/pg
2.7 reverslons/pg
NC
Vehicle was OHSO
Vehicle was DHSO
S-9 harvested from
livers of rats.
yellowtatl fish and/or
Japanese clams; tested
In spores rather than
negative stage
Reference
Takemura and
SMrnliu. 1978
Ho et al..
1979
Brooks and
Dean. 1981
Shlmliu
et al.. 1982
HcCann et al. ,
1975
tchlnotsubo
et al.. 1981
Hatsushlma
et al.. 1981
Thomson, 1981
Kada, 1981
-------
TABLE 6-5 (cont.)
^^
f\J
0»
-w
0.
1
CO
us
i
07/23/90
Assay
Yeast
mutation
Nilotic gene
conversion
Nilotic
aneuploldy
Sex-linked
recessive
lethal
Unscheduled
DMA synthesis
Unscheduled
DNA synthesis
Unscheduled
ONA synthesis
Unscheduled
DNA synthesis
Unscheduled
ONA synthesis
Unscheduled ONA
synthesis and
cytotoxlcl ty
Chromosomal
aberrations
SCE
SCE
Cell trans-
formation
Indicator/Organism
NR
Saccharomyces
cercvlslae J01
S. cerevtslae
D6
Orosophlla
melanogaster
rabbit
hepatocytes
rat
hepatocytes
rat
hepatocytes
mouse
hepatocytes
hamster
hepatocytes
rat
heapatocytes
human
leucocytes
Chinese
hamster
ovary
Syrian
hamster
embryo/SA7
Application Concentration
or Dose
NR NR
prelncubatlon 10 pg
plate
Incorporation
prelncubatlon S \iq
plate
Incorporation
NR NR
18-hour 10~« to 10~« N
Incubation
20-hour 10"» to 10"" H
Incubation
18-hour 10"« to 5x10"« H
Incubation
18-hour 10~« to 5xl(T4 M
Incubation
18-hour 10~» to 5xlO~« H
Incubation
2- to 18-hour 10'« to 10~» M
Incubation
NR NR
26-hour NR
liquid
Incubation
2- or 18-hour 5-20 vg/mt
liquid
Incubation
Activating
System
none
mlcrosomal
S-9
S-9
NA
NA
NA
NA
NA
NA
NA
t
S-9
NA
Response Comment
* NC
+
f Vehicle was DMSO;
concentration was LEO
t Concentration was LEO
NC
t Weak response
4 NC
* NC
+ NC
« NC
» Cells monitored for
release of LDH and
GOT
NC
NC
* NC
Reference
Ho et al.,
1979
Sharp and
Parry. 1981
Parry and
Sharp, 1981
Ho et al..
1979
McQueen
et al.. 1983
Mori et al..
1988
McQueen
et al.s 1981
McQueen
et al.. 1981
McQueen
et al.. 1981
McQueen and
Williams. 1982
Ho et al..
1979
Galloway
et al.. 1985
Casto. 1980
-------
TABLE 6-5 (cent.)
0
CO
S Assay
Cell trans-
formation
Cell trans-
formation
Indicator/Organism
Syrian
hamster
kidney
Balb/3T3
Application Concentration Activating Response -.Comment
or Dose System
liquid NR S-9 * NC
Incubation
3-day liquid 0.01-1.0 pg/ml NA » NC
Incubation
Reference
Purchase
et al.. 1978
Dunkel et al..
19B1
NR =- Not reported; NC = no comment; NT = not tested
o
i
CO
>v
o
-------
Hamster cells were also more sensitive to ONA breakage by 4,4'-methylene-
b1s(2-chloroanH1ne) by the same order of magnitude as that for the L05Q
values.
6.4. DEVELOPMENTAL TOXICITY
Pertinent data regarding the developmental toxldty of 4,4'-methylene-
b1s(2-chloroanH1ne) were not located 1n the available literature cited In
Appendix A.
6.5. REPRODUCTIVE TOXICITY
Pertinent data regarding the other reproductive effects of 4,4'-methyl-
ene-b1s(2-chloroan1!1ne) were not located 1n the available literature dted
In Appendix A.
6.6. SUMMARY
The carcinogenic effects of 4,4'-methylene-b1s(2-chloroan1l1ne) admin-
istered 1n the diets of mice, rats and dogs have been the subject of several
studies. Results have shown that administration In the diet to CD-I mice
produced Increased Incidences of hepatomas 1n females compared with controls
(RussMeld et al., 1975) (see Table 6-1). When given 1n the diet to male
and female Charles River CD rats, 4,4'-methylene-b1s(2-chloroan1l1ne) pro-
/'
duced statistically significant Increased Incidences of lung adenocarcl-
nomas compared with controls (Stula et al., 1975) (see Table 6-2).
Komm1nen1 et al. (1979) reported that dietary 4,4'-methylene-b1s(2-chloro-
anlUne) given to male Charles River Sprague-Dawley rats for 18 months
followed by 6 months of observation led to Increased Incidences of pulmonary
adenomas and adenocarclnomas, mammary adenocarclnomas, Zymbal gland carcino-
mas and hepatocellular carcinomas compared with controls (see Table 6-3).
Four of five female beagle dogs given 4,4'-methylene-b1s(2-chloroan1l1ne) in
gelatin capsules for <9 years developed urinary bladder tumors while no
control dogs developed such tumors (Stula et al., 1977) (see Table 6-4).
0237d -41- 07/23/90
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4f4'-Methylene-b1s(2-chloroannine) , been shown to be mutagenlc In
the Salmonella reverse mutation assay, w';- and without mlcrosomal activa-
tion (Takemura and Shlmlzu, 1978; Ho et al., 1979; Sh1m1zu et al., 1982;
McCann et al., 1975), 1n an E_. coll DNA repair assay with mlcrosomal activa-
tion (Ichlnotsubo et al., 1981) and 1n a yeast mutation assay with and
without activation (Ho et al., 1979). It failed to Induce sex-linked
recessive lethal mutations 1n DrosophUa melanogaster (Ho et al., 1979), and
1t did not produce chromatld aberrations or SCE 1n Chinese hamster ovary
cells or human leucocytes (Galloway et al., 1985; Ho et al., 1979), (see
Table 6-5).
Occupational exposure to 4,4'-methylene-b1s(2-chloroan1l1ne) has been
associated with a reversible form of hematuMa, but exposures were not
precisely quantified (Mastromatteo, 1965). Signs of systemic toxldty,
which was due to oral administration of 4,4'-methylene-b1s(2-chloroan1l1ne)
1n animals, Included liver Injury 1n female beagle dogs administered 7.3
mg/kg/day for 9 years (Stula et al., 1977), high mortality 1n female CD-I
mice at dietary concentrations of 2000 ppm for 18 months followed by 6
months on a normal diet (Russfleld et al., 1975), decreased body weight
gains 1n CD-I male rats administered 500 or 1000 ppm for 18 months followed
by 6 months of observation (Russfleld et al., 1975), decreased survival time
1n CR Sprague-Dawley rats administered 4,4'-methylene-b1s(2-chloroan1l1ne)
1n the diet at 0, 250, 500, and 1000 ppm for 18 months and observed for 6
months (Kommlnenl et al., 1979), and liver effects Including hepatocytome-
galy, fatty change, necrosis, bile duct proliferation and flbrosls In
Charles River CD rats administered 1000 ppm 1n the diets for <2 years (Stula
et al., 1975).
0237d -42- 07/23/90
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lH vitro studies with rat liver cells (S1lk et al., 1989), and dog and
human bladder explant cultures (Stoner et al., 1987) have led to the
Isolation of 4,4'-methylene-b1s(2-chloroan1l1ne)-DNA adducts, suggesting
that 4,4'-methy1ene-b1s(2-chloroan111ne) can bind to DNA 1n these species.
Three of the adducts were common to dog and human cell cultures.
0237d -43- 07/23/90
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7. EXISTING GUIDELINES . /.HOARDS
7.1. HUMAN
The ACGIH (1988) adopted a TLV-TWA of 0.02 ppm (-0.22 mg/m3).
Provided that skin contact 1s avoided, this value win probably prevent
systemic poisoning (ACGIH, 1986). Because 4,4'-methylene-b1s(2-chloro-
anlllne) may cause cancer In humans, probably In the liver or bladder,
4,4'-methylene-b1s(2-chloroan1l1ne) 1s also designated an Industrial
substance suspect of carcinogenic potential to humans. OSHA (1989) estab-
lished, an 8-hour TWA of 0.02 ppm with a skin notation as a final rule for
occupational exposure to 4,4'-methylene-b1s(2-chloroan1l1ne). This regula-
tion Is Intended to protect against material health Impairments and bladder
cancer.
It was recommended that 4,4'-methylene-b1s(2-chloroan1l1ne) exposure to
employees In the workplace be limited to 3 yg/m3 determined as a TWA
concentration for a <10-hour workshlft, 40-hour workweek, over a working
lifetime (NIOSH, 1978).
7.2. AQUATIC
Guidelines and standards for the protection of aquatic life from
exposure to 4,4-methylene-b1s(2-chloroan1l1ne) were not located 1n the
available literature cited In Appendix A.
0237d -44- 07/23/90
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8. RISK ASSESSMENT
8.1. CARCINOGENICITY
8.1.1. Inhalation. Unch et al. (1971) reported no Increase 1n the Inci-
dence of tumors In a cohort of 31 workers exposed to 4,4'-methylene-b1s-
(2-chloroan1l1ne) 1n a manufacturing plant compared wHh 31 nonexposed
workers 1n the same plant. NIOSH (1986, 1987), however, located two cases
of tumor of the urinary bladder 1n preliminary studies of a cohort of 37Q
exposed workers who were screened with a urine cytology examination. The
tumors were not diagnosed by the cytologlc examination but by cystoscopy
performed because of Intermittent hematurla (the first case) or because of
high exposure (the second case).
8.1.2. Oral. Russfleld et al. (1975) administered 4,4'-methylene-b1s-
(2-chloroan1l1ne) In the diets to Charles River CD-I mice of both sexes and
Charles River CD-I male rats. Groups of 25 rats or 25 male and 25 female
mice were administered a commercial diet supplemented with concentrations of
500 and 1000 ppm (rats) or 1000 and 2000 ppm (mice) 4,4'-methylene-b1s-
(2-chloroan1l1ne). Results (see Table 6-1) Indicate that treated female
mice had a statistically Increased Incidence of hepatomas, compared with
controls. The authors Indicated that the difference between the Incidence
of tumors 1n the treated groups of rats and the Incidence of tumors In the
controls was not statistically significant.
Stula et al. (1975) studied the effects of 4,4'-methylene-b1s(2-chloro-
anHlne) on tumor formation In groups of 50 male and 50 female Charles River
CD rats when administered at concentrations of 0 or 1000 ppm 1n the diet for
<2 years. Results (see Table 6-2) showed that treated rats had a
significantly higher Incidence of lung adenocardnomas than controls. Liver
tumors were also observed 1n treated rats of both sexes at Incidences
0237d -45- 07/23/90
-------
greater than 1n controls; however, th erence was not statl: ocally
significant. A study by Kommlnenl et >1979) assessed the effect of
dietary 4,4l-methylene-b1s(2-chloroan1l1np; .r; tumor formation 1n Charles
River Sprague-Dawley male rats. Increased Incidences of pulmonary adeno-
cardnomas, mammary adenocarclnomas, Zymbal gland carcinomas and hepato-
cellular carcinomas were seen (see Table 6-3); they were attributed to
administration of 4,4'-methylene-b1s(2-chloroan1l1ne). Stula et al. (1977)
administered 4,4'-methylene-b1s(2-chloroan1l1ne) In gelatin capsules to a
group -of 6 female purebred beagle dogs, 3 days/week for 6 weeks and then 5
days/week for <9 years and reported that 4/5 treated dogs, but none of the
controls, had carcinomas of the urinary bladder (see Table 6-4).
8.1.3. Other Routes. 4,4'-Methylene-b1s(2-chloroan1l1ne) In saline
administered to rats by subcutaneous Injections was associated with the
development of liver cell carcinomas and primary lung carcinomas (Stelnhoff
and Grundmann, 1971).
8.1.4. Weight of Evidence. The small cohort study by L1nch et al. (1971)
and the preliminary reports by NIOSH (1986, 1987) were lacking sufficient
data to evaluate the cardnogenlcHy of 4,4'-methylene-b1s(2-chloroan1l1ne)
to humans. Sufficient evidence exists regarding the cardnogenlcHy of
4,4'-methylene-b1s(2-chloroan1l1ne) when administered orally to rats, mice
and dogs, and when administered subcutaneously to rats. According to the
U.S. EPA (1986b) classification scheme, 4,4'-methylene-b1s(2-chloroan1l1ne)
can be placed In U.S. EPA Group B2: probable cardnogenlcHy for humans.
8.1.5. Quantitative Risk Estimates.
8.1.5.1. INHALATION -- Pertinent data were not located regarding the
cardnogenlcHy of Inhalation exposure of animals to 4,4'-'methylene-b1s-
(2-chloroan1l1ne). The occupational studies by NIOSH (1986, 1987) and Linen
0237d -46- 07/23/90
-------
et al. (1971) are not useful for cancer risk assessment. Dietary studies
have reported the development of liver tumors 1n female mice and male rats
(Russfleld et al., 1975; Komm1nen1 et al., 1979), lung tumors In male and
female rats (Stula et al., 1975; Kommlnenl et al., 1979) and urinary bladder
tumors in female beagle dogs (Stula et al., 1977). Liver and lung tumors
have been reported 1n rats following subcutaneous administration (Stelnhoff
and Grundmann, 1971). These data suggest that 4,4'-methylene-b1s(2-chloro-
anlUne) may be carcinogenic by any route of exposure, provided that distri-
bution of the proximate carcinogen to the liver, lung and bladder occurs.
Pharmacoklnetlc data (Section 5.1.) Indicate that distribution to these
organs does occur. Thus, It Is appropriate to adopt the q * of l.SxlCT1
(mg/kg/day)"1 derived -for oral exposure (see below) as the q,* for
Inhalation exposure as well.
In estimating the concentration of 4,4'-methylene-b1s(2-chloroan1l1ne)
In air associated with specific levels of Increased risk of cancer, the
ratio of the extent of absorption from the respiratory tract to that from
the gastrointestinal tract must be adjusted. Pharmacoklnetlc data suggest
that absorption from the gastrointestinal tract 1s rapid and complete. Data
are lacking regarding the extent of absorption from the respiratory tract.
A default value of 50% 1s assumed, resulting 1n a resp1ratory:gastro1ntes-
tlnal absorption ratio of 0.5. By applying the adjustment factor of 0.5 and
by assuming that humans weigh 70 kg and Inhale 20 mVday, 1t 1s estimated
that an air concentration of 5.4xlO~4 mg/m3 would be associated with an
Increased cancer risk of lx!0~s. Concentrations In air of 5.4xlO"5
mg/m3 and 5.4xlO~6 mg/m3 are associated with Increased cancer risks of
IxlO'6 and IxlO"7, respectively.
0237d -47- 07/23/90
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8.1.5.2. ORAL ~ ;»v'rif1eld et al. : 5) reported a statistically
significant Increase 1n the Incidence of , tomas 1n female Charles River
CD-I mice fed diets containing 4,4'-methylcl e-b1s(2-chloroanH1ne) for 18
months compared with controls. Stula et al. (1975) observed statistically
significant Increases compared with controls 1n the Incidence of lung adeno-
carclnomas In male and female Charles River CD rats fed 4,4'-methylene-b1s-
(2-chloroan1l1ne) 1n the diet for <2 years. Kommlnenl et al. (1979)
reported that the Incidences of lung adenocardnomas and all primary
neoplasms were significantly Increased 1n Charles River Sprague-Dawley rats
that were administered 4,4'-methylene-b1s(2-chloroan1l1ne) In the diet for
18 months. Slope factors for oral exposure to 4,4'-methylene-b1s(2-chloro-
anlllne) were determined from data from each of these studies using the
GLOBAL86 program for the multistage model designed by Howe et al (1986).
The data and calculations for the derivations are presented In Appendix B.
A human q.j* of 8.9xlO~2 (mg/kg/day)'1 was calculated from the
RussHeld et al. (1975) mouse study; a human q^ of 1.2XKT1 (mg/kg/
day)"1 was calculated from the Stula et al. (1975) report and a human
q.j* of l.SxlO"1 (mg/kg/day)'1 was determined from the Kommlnenl et al.
(1979) study. The q * values for lung tumors In the two rat studies are
similar. Because these studies used larger numbers of animals than the
mouse study and early mortality was lower In the two rat studies than 1n the
mouse study, It 1s more appropriate to use the rat studies for estimation of
cancer potency. The most appropriate risk estimate for 4,4'-methylene-b1s-
(2-chloroan1l1ne) 1s derived by computing the geometric average of the two
q.j* values for lung tumors In rats. The geometric mean of 0.12 and 0.15
1s 0.13, the human q * for oral exposure to 4,4'-methylene-b1s(2-chloro-
anlUne) 1s, therefore, calculated as l.SxlO"1 (mg/kg/day)'1. From the
0237d -48- 07/23/90
-------
q-!* of 1.3x10 » (mg/kg/day)"1, H Is estimated that a concentration of
2.7xlO~3 mg/i 1n drinking water 1s associated with Increased cancer risk
to humans of lxlO~s. This estimate Is based on the assumption that humans
weigh 70 kg and drink 2 l water/day. Drinking water concentrations of
2.7xlO"4 and 2.7xlO~5 mg/i are associated with Increased cancer risks
of lx!0~« and IxlO'7, respectively.
8.2. SYSTEMIC TOXICITY
8.2.1. Inhalation Exposure.
8.2.1.1. LESS THAN LIFETIME (SUBCHRONIC) -- Pertinent data regarding
the toxldty of subchronlc Inhalation exposure to 4,4'-methylene-b1s-
(2-chloroanlllne) were not located In the available literature cited 1n
Appendix A; data are Insufficient for derivation of an RfD for subchronlc
Inhalation exposure.
8.2.1.2. CHRONIC Llnch et al. (1971) reported no health effects on
workers exposed to 4,4'-methylene-b1s(2-chloroan1l1ne). Mastromatteo
(1965), however, associated occupational exposure with hematuMa. Exposures
were not sufficiently quantified In either study for use In derivation of an
RfD for chronic Inhalation exposure.
8.2.2. Oral Exposure.
8.2.2.1. LESS THAN LIFETIME (SUBCHRONIC) -- Pertinent data regarding
toxldty of subchronlc oral exposure to 4,4'-methylene-b1s(2-chloroan1l1ne)
were not located 1n the available literature dted In Appendix A. Data are
available for derivation of a chronic RfD and this value can be adopted as
sufficiently protective for subchronlc exposure. The value of 0.0007 mg/kg/
day 1s adopted as the RfD for subchronlc oral exposure to 4,4'-methylene-
b1s(2-chloroan1Hne). The confidence 1n this value 1s low.
0237d -49- 07/23/90
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8.2.2.2. CHRONIC ~ Data \ ... -our different chronic s'.-,.>.$$ using
three different species are available for the determination of a chronic
oral RfD for 4,4'-methylene-b1s(2-chloroan1l1ne). Stula et al. (1977) (Rec.
#1, Appendix D) reported liver nodular hyperplasla, Increased GPT activity
and folUcular cystitis In female beagle dogs (n=6) that received an average
dose of 7.3 mg 4,4'-methylene-b1s(2-chloroan1l1ne)/kg/day In gelatin
capsules for 9 years. These effects Indicated liver Injury and bladder
Inflammation 1n the dogs. There appeared to be no treatment-related effects
on mortality and body weights. The weaknesses of the study are that only
female dogs were used, only six dogs were given the compound and because
only one dose was administered; therefore, no dose-response relationships
could be determined.
Charles River CD-I mice were given 0, 1000 (Rec. #2, Appendix 0) or 2000
ppm (Rec. #3, Appendix D) for 18 months and observed for an additional 6
months In a study by Russfleld et al. (1975). Assuming a food factor of
0.13 kg food/kg bw/day, these dietary concentrations correspond to doses of
0, 130 and 260 mg/kg/day, respectively. The only adverse effect observed
was Increased mortality 1n the female mice at the higher dose. The authors
reported that the Incidence and Intensity of amyloldosis was reduced In
treated mice compared with controls. These Investigators also fed rats
diets containing 0, 500 (Rec. #6, Appendix D) or 1000 ppm for 18 months and
observed them for an additional 6 months. Assuming a food factor for rats
of 0.05 kg food/kg bw/day (U.S. EPA, 1980), these dietary concentrations
correspond to doses of 0, 25 and 50 mg/kg/day. Treated rats exhibited
decreased mean body weights compared with controls throughout the study.
This effect was dose-related. There were no effects on mortality. The
Russfleld et al. (1975) study using mice and rats Is not considered further
0237d -50- 07/23/90
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for risk assessment because a 6-month recovery period was permitted after
exposure, during which reversible adverse effects could have been repaired,
thereby escaping detection at necropsy and hlstopathologkal examination.
Kommlnenl et al. (1979) fed male rats diets containing 0, 250 (Rec. #4,
Appendix D), 500 (Rec. #5, Appendix D) or 1000 ppm 1n the diets for 18
months. They were then observed for an additional 6 months. As calculated
above, corresponding doses can be estimated at 0, 12.5, 25 and 50 mg/kg/day,
respectively. Significantly reduced survival was reported at 500 and 1000
ppm. 'Body weight gain was lower than controls. The posttreatment recovery
period precludes considering this study further for risk assessment.
Stula et al. (1975) reported hepatocytomegaly, fatty liver change, liver
necrosis, bile duct proliferation and flbrosls 1n rats fed a diet containing
1000 ppm for 2 years (Rec. #7, Appendix D). Survival also appeared to be
reduced In treated rats of both sexes. The 1000 ppm level (corresponding to
50 mg/kg/day as computed above) 1s considered a PEL and cannot be used In
risk assessment.
Comparison of the studies Indicates that dogs are more sensitive to the
systemic effects of 4,4'-methylene-b1s(2-chloroan1l1ne) than rats or mice
because they exhibit adverse effects at lower dose levels. It 1s possible
that adverse effects would have been Identified In rats and mice at lower
levels 1f a 6-month recovery period had not been provided. The lowest dose
at which an adverse effect was noted was 7.3 mg/kg/day In female beagle dogs
(Stula et al., 1977), which 1s considered to be a LOAEL for systemic
toxldty (Rec. #1, Appendix D). The RfO for systemic toxldty 1s calculated
by dividing the LOAEL by an uncertainty factor of 10,000 (10 to reflect the
deficiencies of the data base, 10 to extrapolate from dogs to humans, 10 to
provide additional protection for more sensitive Individuals and 10 for
estimation of .a NOAEL from a LOAEL) and Is 0.0007 mg/kg/day.
0237d -51- 07/23/90
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Conflden: a the key study Is low because : ;o sizes were small, dogs
of only one sex were used and only one dose was given, which precluded
locating the threshold for adverse effects. Confidence In the data base 1s
low. The other chronic studies available 1n rats and mice provide limited
Information because of the recovery period after exposure (Russfleld et
al.,1975; Kommlnen! et al., 1979) or because only one dose was given (Stula
et al., 1975). Furthermore, there 1s no Information regarding the reproduc-
tive or developmental effects of 4,4'-methylene-b1s(2-chloroan1l1ne).
Confidence 1n the RfO, therefore, 1s low.
0237d -52- 07/23/90
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9. REPORTABLE QUANTITIES
9.1. BASED ON SYSTEMIC TOXICITY
The systemic toxldty of 4,4'-methylene-b1s(2-chloroannine) was dis-
cussed 1n Chapter 6. For each study considered for computation of candidate
CSs, the lowest doses associated with the effects reported are summarized 1n
Table 9-1. Effects noted In chronic exposure studies using female dogs
Included liver Injury and urinary bladder Inflammation (Stula et al., 1977);
the effect seen 1n a chronic study with female mice was Increased mortality
compared with controls (RussMeld et al., 1975); effects observed In male
rats Included dose-related decreases In body weights (Russfleld et al.,
1975) and decreased body weights and a dose-related decrease In mean
survival times (Kommlnenl et al., 1979); effects reported 1n rats of both
sexes were decreased survival and signs of liver Injury, Including hepato-
cytomegaly, fatty change, necrosis, bile duct proliferation and flbrosls
(Stula et al., 1975). More severe effects may have been Identified In the
studies by Russfleld et al. (1975) and Kommlnenl et al. (1979) had the rats
and mice been examined at the termination of the exposure period rather than
after a 6-month recovery period.
Table 9-2 presents candldate'CSs for the effects presented 1n Table 9-1.
CSs are calculated only for the lowest human equivalent dose associated with
each effect In Table 9-1. The highest CS, 17.86, was calculated for
significantly Increased mortality compared with controls In male rats
exposed to a dietary concentration of 500 ppm for 18 months (Kommlnenl et
al., 1979). The CS of 17.86 corresponding to an RQ of 1000 1s chosen to
represent the chronic (noncancer) toxldty of 4,4'-methylene-b1s(2-chloro-
anmne) (Table 9-3).
0237d -53- 07/23/90
-------
0
CO
a.
Species/Strain
Dog/beagle
Mouse/Charles
i River CD-I
Rat/Charles
River CD-I
Rat/Sprague-
Dawley
Rat/Charles
River CD
TABLE 9-1
Oral Toxlclty Summary for 4.4'-Methy1ene-b1s(2-Chloroan111ne)
No. at Average Vehicle/ Transformed
Sex Start Weight Physical Purity Exposure Animal Dose
(kg) State (mg/kg/day)
F 6 10. 4b dietary/ 90X 100 mg/day, 7.3
capsule 3 days/week for
6 weeks, then
5 days/week for
9 years (avg. dose
.7.3 mg/kg/day)
F 25 0.03C dietary 97X 2000 ppm for 260. Od
18 months
N 25 0.35C dietary 97X 500 ppm for 25. Od
18 months
N 100 0.35C dietary NR 500 ppm for 25. Od
18 months
H/F 50/sex 0.25C dietary 95X 1000 ppm for 50. Od
2 years
Equivalent
Human Dose3 Response
(mg/kg/day)
3.87 Liver Injury, bladder
Inflammation
19.60 Increased mortality
compared with controls
4.27 Decreased mean body
weights compared with
controls
4.27 Increased mortality
compared with controls
(p<0.01)
7.64 Decreased survival;
liver Injury
Reference
Stula
et al.,
1977
Russfleld
et al., 1975
RussMeld
et >.!. '
Ki-
el *!..
Stula
et al..
19/9
1975
Calculation: transformed animal dosage (mg/kg/day) x [animal body weight (kg)/reference human body weight (70 kg)]1/3
bEsMamted from data provided by Investigators
(Reference body weight (U.S. EPA. 1980)
dfood factors used: rats. 0.05 kg food/kg bw/day; mice, 0.13 kg food/kg bw/day (U.S. EPA. 1980)
NR - Not reported
uo
O
-------
o
OJ
to
TABLE 9-2
Composite Scores for 4,4'-Methylene-b1s(2-Chloroan1l1ne)
1
en
en
0
j
co
\
0
Chronic
Species Animal Dose Human MED RV,j Effect
(mg/kg/day) (mg/day)
Dog 7.3 270.65 1.85 Liver Injury; bladder
Inflammation
Rat 25.0 299.25 1.79 Decreased mean body
weights compared with
controls
Rat 25.0 299.25 U79 Increased mortality
compared with controls
'Although Equivalent Human Dose In Table 9-1 and chronic human MED,
as two digits past the decimal, no rounding was performed In the
transformed animal dose and ending with CS.
RVe CS* RQ Reference
5 9.26 1000 Stula
et al., 1977
4 7.14 1000 Russfleld
et al., 1975
10 17.86 1000 Komm1nen1
et al., 1979
RV(j and CS In Table 9-2 are written
chain of calculations starting with
-------
TABLE 9-3
4,4'-Methylene-b1s(2-Chloroan1l1ne)
Minimum Effective Dose (MED) and Reportable Quantity (RQ)
Route: oral, diet
Species/Sex: rat/male
Dose*: 299.25 mg/day
Duration: 18 months
Effect: Increased mortality compared with controls
RVd: 1.79
RVe: 10
CS: 17.86
RQ: 1000
Reference: Kommlnenl et al., 1979
*EquWalent human dose
0237d -56- 07/23/90
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9.2. BASED ON CARCINOGENICITY
The cardnogenkHy of 4,4'-methylene-b1s(2-chloroannine) was discussed
In Chapter 6. In the only data available regarding the cardnogenlcHy of
4,4'-methylene-b1s(2-chloroannine) In humans, Unch et al. (1971) reported
no effects In an occupational study Involving 31 exposed and 31 nonexposed
workers, but NIOSH (1986, 1987) located two cases of urinary bladder tumors
1n 370 workers 1n a 4,4'-methylene-b1s(2-chloran1l1ne) manufacturing plant.
Studies with laboratory animals associate bladder tumors In dogs (Stula
et al., 1977), hepatomas In mice (Russfleld et al., 1975) and lung tumors 1n
rats (Stula et al., 1975; Komm1nen1 et al., 1979) with oral exposure to
4,4'-methylene-b1s(2-chloroan1l1ne). Subcutaneous administration of
4,4'-methylene-b1s(2-chloroan1l1ne) to rats resulted In Increased Incidences
of primary lung carcinomas and liver cell carcinomas compared with controls
(Stelnhoff and Grundmann, 1971).
The above animal studies provide positive evidence for the carclnogen-
1c1ty of 4,4'-methylene-b1s(2-chloroan1l1ne). 4,4'-Methylene-b1s(2-chloro-
anHlne) was placed In CAG Group B2: probable human carcinogen.
Using the data presented In Table 9-4 and Appendix B and the multistage
model by Howe et al. (1986), a human F factor of 0.79 (mg/kg/day)~a was
estimated, which corresponds to a Potency Group of 3. Potency Group 3
compounds In U.S. EPA Group B are assigned a "low" hazard ranking, which
corresponds to an RQ of 100 for cardnogenlcHy.'
0237d -57- 07/23/90
-------
TABLE 9-4
Derivation of Potency Factor (F) for 4,4'-Methylene-b1s(2-chloroan1l1ne)
References: Stula et al., 1975; Komm1nen1 et al., 1979
Specles/straln/sex: Charles River CD/male and female; Charles River Sprague-
Dawley/male
Route/vehicle: oral/diet
Length of exposure (le) = 2 years; 18 months
Length of experiment (LE) = 2 years; 2 years
Llfespan of animal (L) = 2 years
Body weight = see Appendix B-2 and B-3
Tumor type: adenocardnoma; adenocardnoma and all lung neoplasms
Adjusted 1/ED10 (F factor): 0.79 (mg/kg/day)'1
Exposure Transformed Dose Incidence
(mg/kg/day) No. Responding/No. Tested
see Appendix B see Appendix B see Appendix B
0237d -58- 07/23/90
-------
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Yoneyama, K. and F. Matsumura. 1984. Mlcroblal metabolism 4,4'-methylene-
b1s(2-chloroan1!1ne). Arch. Environ. Contam. Toxlcol. 13: 501-507.
0237d -70- 07/23/90
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APPENDIX A
LITERATURE SEARCHED
This HEED Is based on data Identified by computerized literature
searches of the following:
CHEMLINE
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)
HSDB
SCISEARCH
Federal Research In Progress
These searches were conducted In April, 1989, and the following secondary
sources were reviewed:
AC6IH (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 Hyglenlsts).
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. 28. John Wiley and
Sons, NY. p. 2879-3816.
0237d -71- 07/23/90
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Clayton, G.D. and F.E. Clayton, , 1982. Patty's Industrial
Hygiene and Toxicology, 3rd rev. Vol. 2C. John Wiley and
Sons, NY. p. 3817-5112.
Grayson, H. and D. Eckroth, Ed. IvH K34. 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 1n the Special Review
Program, Registration Standards Program and the Data Call 1n
Programs. Registration Standards and the Data Call In Programs.
Office of Pesticide Programs, Washington, DC.
USITC (U.S. International Trade Commission). 1986. Synthetic
Organic Chemicals. U.S. Production and Sales, 1985, USI1C Publ.
1892, Washington, DC.
Verschueren, K. 1983. Handbook of Environmental Data on Organic
Chemicals, 2nd ed. Van Nostrand Relnhold Co., NY.
Wlndholz, M., Ed. 1983. The Merck Index, 10th ed. Merck and Co.,
Inc., Rahway, NJ.
Worthing, C.R. and S.B. Walker, Ed. 1983. The Pesticide Manual.
British Crop Protection Council. 695 p.
0237d -72- 07/23/90
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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. Mnley. 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
Spedes. 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.
0237d -73- 07/23/90
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APPEND,
Cancer Data Sheet for DeMvat . a q-j* Using G1oba!86
Reference: Russfleld et al., 1975
Specles/straln/sex: mouse/Charles River CD-1/female
Route/vehicle: oral/food
Length of exposure (le) = 18 months
Length of experiment (LE) = 24 months
Llfespan of animal (L) = 24 months
Body weight = 0.03 (assumed)3
Tumor site and type: liver hepatomas
Purity: 97%
Exposure
(ppm)
0
1000
2000
Transformed Doseb
(mg/kg/day)
0
97.5
195
Incidence
No. Responding/No.
0/20
9/21
7/14
Tested
Reference value from U.S. EPA (1980)
bEst1mated by applying a reference food Intake factor for mice of 0.13 kg
diet/kg bw/day (US. EPA, 1980) and multiplying the result by 18/24 months
to expand to continuous exposure
Unadjusted q-\* = 6.72965xlO~3 (mg/kg/day)"1
Human q? = 8.9xlO"2 (mg/kg/day)"1
Polynomial degree selection procedure and test selected by program using
Monte Carlo test.
0237d -74- 07/23/90
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APPENDIX B-2
Cancer Data Sheet for Derivation of a q-|* Using Global 86
Reference: Stula et al., 1975
Specles/straln/sex: Rat/Charles River CD-1/male and female3
Route/vehicle: oral/food
Length of exposure (le) = 24 months
Length of experiment (LE) = 24 months
LUespan of animal (L) = 24 months
Body weight = 0.35 (assumed)13
Tumor site and type: lung adenocardnoma
Purity: 95%
Exposure
(ppm)
0
1000
Transformed Dose
(mg/kg/day)
0
50
Incidence
No. Responding/No.
0/88
48/88
Tested
aData from males and females were combined because neither gender appeared
to be more sensitive.
bReference value from U.S. EPA (1980)
Estimated by applying a reference food Intake factor of 0.05 kg diet/kg
bw/day (U.S. EPA, 1980)
Unadjusted q]* = 1.99485xlO~2 (mg/kg/day)"1
Human q-|* = 1.2X10"1 (mg/kg/day)'1
Polynomial degree selection procedure and test selected by programming using
Monte Carlo test.
0237d -75- 07/23/90
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APP -3
Cancer Data Sheet for DeMv, :<^ of a q-|* Using Global 86
Reference: Komm1nen1 et al., 1979
Specles/straln/sex: rat/Charles River Sprague-Dawley/male
Route/vehicle: oral/food
Length of exposure (le) = 18 months
Length of experiment (LE) = 24 months
Llfespan of animal (L) = 24 months
Body weight = 0.59 for 0, 250 and 500 ppm groups; 0.52 for 1000 ppm group3
Tumor site and type: lung, all primary neoplasms
Purity: NR
Exposure
0
250
500
1000
Transformed Dose''
(mg/kg/day)
0
9.4
18.8
37.5
Equivalent
Human Dosec
(mg/kg/day)
0
1.91
3.83
7.32
Incidence
No. Responding/No. Tested
1/100
23/100
28/75
35/50
Estimated from graphic data provided by Investigators
bEst1mated by applying a reference food Intake factor of 0.05 kg diet/kg
body weight (U.S. EPA, 1980) and multiplying the result by 18/24 months to
expand to continuous exposure
cEst1mated by mul1ply1ng the transformed animal dose by the cube root of
the ratio of the rat body weight/human reference body weight of 70 kg
Human q-j* = 1.5X10"1 (mg/kg/day)'1
Polynomial degree selection procedure and test selected by -program using
Monte Carlo test.
NR = Not reported
0237d -76- 07/23/90
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Summary Table for MOCA
o
r\j
~j
O.
Inhalation Exposure
Subchronlc
Chronic
CardnogenUHy
Oral Exposure
Subchronlc
Species Exposure Effect
ID ID ID
ID ID ID
rat oral, diet lung tumors
see Appendix B
dog 100 nig/day. 3 days/week liver Injury.
RfD or. qi* Reference
ID ID
ID ID
1.3xlO~» (mg/kg/day)"* Stula et al.. 1975
Kommlnenl et al.,
1979
0.0007 mg/kg/day Stula et al..
1977
Chronic
Carclnogenlclty
for 3 weeks, then 5
days/week for 9 years
(7.3 mg/kg/day); LOAEL
dog 100 fng/day. 3 days/week
for 3 weeks, then 5
days/week for 9 years
(7.3 mg/kg/day); LOAEL
rat oral, diet
see Appendix B
bladder Inflammation
liver Injury,
bladder Inflammation
lung tumors
0.0007 mg/kg/day
1.3xlO-» (mg/kg/day)'1
Stula et al.. 1977
Stula et al.. 197S
Kommlnenl et al.,
1979
REPORTABLE QUANTITIES
Based on chronic toxlclty:
Based on carclnogenlclty:
1000
100
Kommlnenl et al.,
1979
Stula et al.. 1975
Kommlnenl et al.,
1979
ro
CO
ID - Insufficient data
UD
O
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APPENDIX D
DOSE/DURATION RESPONSE GRAPHS FOR EXPOSURE T' '..4-HETHYLENE-BIS-
(2-CHLOROANILINE)
D.I. DISCUSSION
Dose/duration-response graphs for oral exposure to 4,4'-methylene-b1s-
(2-chloroan111ne) generated by the method of Crockett et al. (1985) using
the computer software by Durkln and Meylan (1988) developed under contract
to ECAO-C1ndnnat1 are presented 1n Figures D-l and D-2. Data used to
genera.te these graphs are presented 1n Figure D-2. In the generation of
these figures, all responses are classified as adverse (FEl, AEL or LOAEL)
or nonadverse (NOEL or NOAEL) for plotting. For oral exposure, the ordlnate
expresses dosage as human equivalent dose. The animal dosage 1n mg/kg/day
Is multiplied by the cube root of the ratio of the animal:human body weight
to adjust for species differences 1n basal metabolic rate (Mantel and
Schnelderman, 1975). The result 1s then multiplied by 70 kg, the reference
human body weight, to express the human equivalent dose as mg/day for a 70
kg human.
The boundary for adverse effects (solid line) Is drawn by Identifying
the lowest adverse effect dose or concentration at the shortest duration of
exposure at which an adverse effect occurred. From this point an Infinite
line Is extended upward parallel to the dose axis. The starting point Is
then connected to the lowest adverse effect dose or concentration at the
next longer duration of exposure that has an adverse effect dose or concen-
tration equal to or lower than the previous one. This process 1s continued
to the lowest adverse effect dose or concentration. From this point a line
Is extended to the right parallel to the duration axis. The region of
adverse effects lies above the adverse effects boundary.
0237d -78- 07/23/90
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IBWkJU
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T
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y
X
V
u:
V.
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e 100? -
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c
It
Z
c
x
t aa
t *. \ m'
; \.^
x^ j
t
4
\
\
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i t i * i j i i f i i i t i t i i 1 r t i i i ' F 1
0.001
<0ral
o.ej 0.1
HUMAN E9UIU DURATION (fraction li/>span>
ENVELOP METHOD
Key: F . FEL
L - LOAEL
A . AEL
N . NOEL
Solid line » Adverse Effect Boundary
Dashed line - No Adverse Effects Boundary
FIGURE 0-1
Dose/Duration Response Graph for Oral Exposure to 4,4'-methylene-b1s
(2-Chloroan1l1ne); Envelope Method
0237d
-79-
07/23/90
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\
p
c
fi
1988 -
Z
e
ice
0.0(31
<0r»l Exposure)
F3
n2
F7
6.81 0.1
HUH4N EQUIU DURATION
Key:
F
L
A
N
FEL
LOAEL
AEL
NOEL
Solid line « Adverse Effect Boundary
Dashed line « No Adverse Effects Boundary
FIGURE 0-2
Dose/Duration Response Graph for Oral Exposure to 4,4'-methylene-b1s-
(2-chloroan1l1ne); Censored Data Method
0237d
-80-
07/23/90
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Using the envelope method, the boundary for no adverse effects (dashed
line) 1s drawn by Identifying the highest no adverse effects dose or concen-
tration. From this point, a line parallel to the duration axis Is extended
to the dose or concentration axis. The starting point 1s then connected to
the next lower or equal no adverse effect dose or concentration at a longer
duration of exposure. When this process can no longer be continued, a line
Is dropped parallel to the dose or concentration axis to the duration axis.
The no adverse effects region lies below the no adverse effects boundary.
At both ends of the graph between the adverse effects and no adverse effects
boundaries are regions of ambiguity. The area (1f any) resulting from
Intersection of the adverse effects and no adverse effects boundaries Is
defined as the region of contradiction.
In the censored data method, all no adverse effect points located In the
region of contradiction are dropped from consideration and the no adverse
effect boundary Is redrawn so that H does not Intersect the adverse effects
boundary and no region of contradiction 1s generated. This method results
1n the most conservative definition of the no adverse effects region.
Figure D-l presents the dose/duration-response graph for oral exposure
generated by the envelope method. The adverse effects boundary Is defined
by an LD5Q value In male rats (Rec. #8), a 10-day AEL In rats (Rec. #9)
and the LOAEL for liver and urinary bladder effects In dogs (Rec. #1) that
served as the basis of the RfD value for oral exposure. The only nonadverse
effect levels are a NOAEL In mice (Rec. #2) and a NOEL 1n rats (Rec. #4)
from studies that allowed a 6-month recovery period after exposure was
terminated, which seriously erodes confidence 1n the designation of these
two data points as nonadverse effect levels. The graph generated by the
censored data method Is presented 1n Figure D-2.
0237d -81- 07/23/90
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D.2. DATA USED TO GENERATE CC3E/DURATION-REL GRAPHS
D.2.1. Inhalation Exposure. Inhalation data :r :able for graphing were
not located.
D.2.2. Oral Exposure.
Chemical Name: 4,4'-Methylene-b1s(2-chloroan1l1ne) (MOCA)
CAS Number: 101-14-4
Document Title: Health and Environmental Effects Document for
4,4'-Hethylene b1s(2-ChloroanH1ne)
Document Number: Pending
Document Date: Pending
Document Type: HEED
RECORD #1:
Comment:
Citation:
RECORD #2:
Species: Dogs
Sex: Female
Effect: LOAEL
Route: Capsul
Number Exposed: 5
Number Responses: 3
Type of Effect: DEGEN
SHe of Effect: LIVER
Severity Effect: 5
Dose:
Duration
Duration
5
5
OTHER
OTHER
5
7.300
Exposure: 9.0 years
Observation: 9.0 years
100 mg/day 3 days/week for 6 weeks, 5 days/week for 9
years/liver Injury and urinary bladder Inflammation.
Stula et al., 1977
Species: Mice
Sex: Both
Effect: NOAEL
Route: Food
Dose;
Duration
Duration
130.000
Exposure: 18.0 months
Observation: 24.0 months
Comment:
Citation:
Number Exposed: 50
Number Responses: NR
Type of Effect: HISTO
Site of Effect: BODY
Severity Effect: 1
1000 ppm (doses studied: 1000, 2000 ppm); reduced amyloldosls,
Russfleld et al., 1975
0237d
-82-
07/23/90
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RECORD #3:
Comment:
Citation;
Species:
Sex:
Effect:
Route:
Mice
Female
PEL
Food
Dose: 260.000
Duration Exposure: 18.0 months
Duration Observation: 24.0 months
Number Exposed: 25
Number Responses: 9
Type of Effect: DEATH
SHe of Effect: BODY
Severity Effect: 10
Comment:
Citation:
RECORD #4:
2000 ppm (see previous record), reduced survival In
Russfleld
Species:
Sex:
Effect:
Route:
et al.. 1975
Rats
Male
NOEL
Food
Dose:
Duration Exposure:
Duration Observation:
females.
12.500
540.0 days
720.0 days
Number Exposed:
Number Responses:
Type of Effect:
SHe of Effect:
Severity Effect:
100
0
Comment:
Citation:
RECORD #5:
250 ppm (Doses studied: 250, 500, 1000 ppm).
Kommlnenl
Species:
Sex:
Effect:
Route:
et al., 1979
Rats
Male
FEL '
Food
Dose:
Duration Exposure:
Duration Observation:
25.000
540.0 days
720.0 days
Number Exposed: 75
Number Responses: 65
Type of Effect: DEATH
SHe of Effect: BODY
Severity Effect: 10
500 ppm (see previous record); rats
reduced rate of body weight gain.
Kommlnenl et al., 1979
at 1000 ppm also had
0237d
-83-
07/23/90
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RECORD #6:
Comment:
Citation:
Species:
Sex:
Effect:
Route:
Rats
Male
LOAEL
Food
Dose:
Duration Exposure:
Duration Observation:
25.000
18.0 months
24.0 months
Number Exposed: 25
Number Responses: NR
Type of Effect: WGTDC
SHe of Effect: BODY
Severity Effect: 4
500 ppm (Doses studied: 500, 1000 ppm); dose-related decrease
In terminal body weights.
Russfleld et al.. 1975
RECORD #7:
Species: Rats
Sex: Both
Effect: PEL
Route: Food
Number Exposed:
Number Responses:
Type of Effect:
SHe of Effect:
Severity Effect:
100
NR
DEATH
BODY
10
Dose:
Duration Exposure:
Duration Observation:
100
NR
NECRO
LIVER
6
50.000
2.0 years
2.0 years
Comment: 1000 ppm; other liver effects: hepatocytomegaly,
change, bile duct proliferation and flbrosls
Citation: Stula et al., 1975
fatty
RECORD #8:
Comment:
Citation:
Species: Rats
Sex: Male
Effect: FEL
Route: Oral (NOS
Number Exposed:
Number Responses:
Type of Effect:
SHe of Effect:
Severity Effect:
Lowest LDso reported
Miller and Sherman,
Dose:
Duration Exposure:
Duration Observation:
)
NR
NR
DEATH
BODY
10
In rats.
1965
750.000
1.0 days
1 .0 days
0237d
-84-
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RECORD #9:
Comment:
Citation:
Species:
Sex:
Effect:
Route:
Rats
NR
AEL
Oral
(NOS)
Number Exposed: NR
Number Responses: NR
Type of Effect: WGTDC
Site of Effect: BODY
Severity Effect: 4
Dose: 200.000
Duration Exposure: 10.0 days
Duration Observation: 10.0 days
NR
NR
HEMAT
BLOOD
UNCL
NR
NR
EXCRE
N.S.
UNCL
Cyanosis, pallor, "blood and urine abnormalities.
Relnke, 1963
NR = Not reported
0237d
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