United States
Environmental Protection
Agency
500ECAOCING003
EPA Research and
Development
HEALTH AND ENVIRONMENTAL EFFECTS DOCUMENT
FOR CHLOROANILINES
Prepared for
OFFICE OF SOLID HASTE AND
EMERGENCY RESPONSE
. . . Do^. , Protection
Prepared by S^t£SBm street
Environmental Criteria and AsW's'SRrtfnf°0flice
Office of Health and Environmental Assessment
U.S. Environmental Protection Agency
Cincinnati, OH 45268
DRAFT: DO NOT CITE OR QUOTE
NOTICE
This document Is a preliminary draft. It has not been formally released
by the U.S. Environmental Protection Agency and should not at this stage be
construed to represent Agency policy. It Is being circulated for comments
on Its technical accuracy and policy Implications.
rt»
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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 (HEEDs) are prepared for the
Office of Solid Haste and Emergency Response (OSWER). This document series
1s 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 from 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 1s current up to 8 months previous to the final
draft date listed on the front cover. Final draft document dates (front
cover) reflect the date the document Is sent to the Program Officer (OSWER).
Several quantitative estimates are presented provided sufficient data
are available. For systemic toxicants, these Include Reference doses (RfDs)
for chronic and subchronlc exposures for both the Inhalation and oral
exposures. The subchronlc or partial lifetime RfD, 1s an estimate of an
exposure level that would not be expected to -cause adverse effects when
exposure occurs during a limited time Interval, for example, one 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, RfDs are not estimated. A
carcinogenic potency factor, or q-|* (U.S. EPA, 1980), 1s provided Instead.
These potency estimates are derived for both oral and Inhalation exposures
where possible. In addition, unit risk estimates for air and drinking water
are presented based on Inhalation and oral data, respectively.
Reportable quantities (RQs) based on both chronic toxldty and carclno-
genlclty are derived. The RQ Is used to determine the quantity of a hazar-
dous substance for which notification 1s required 1n the event of a release
as specified under the CERCLA. These two RQs (chronic toxldty and carclno-
genlclty) represent two of six scores developed (the remaining four reflect
1gn1tab1l1ty, reactivity, aquatic toxldty, and acute mammalian toxlclty).
Chemical-specific RQs reflect the lowest of these six primary criteria. The
methodology for chronic toxldty and cancer-based RQs are defined 1n U.S.
EPA, 1983 and 1986a, respectively.
111
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EXECUTIVE SUMMARY
2-Chloroan1l1ne (CAS number 95-51-2) 1s an amber liquid; 3-chloroan1l1ne
(CAS number 108-42-9) 1s a colorless to light amber liquid; and 4-chloroan1-
Une (CAS number 106-47-8) Is a white to pale yellow solid at room tempera-
ture (Hawley, 1981). They are slightly soluble In water and are soluble In
common organic solvents (Hawley, 1981). The 1986 Directory of Chemical
Producers (SRI, 1986) reports that E.I. Dupont 1n Oeepwater, NJ, currently
produces all three chloroanlUne Isomers; The Upjohn Co. 1n North Haven, CT,
produces 2-chloroan1l1ne; First Mississippi Corp. produces 3-ch1oroan1l1ne;
and Monsanto Co. 1n Lullng, LA, produces 4-ch1oroan1l1ne. Domestic produc-
tion volume data for recent years could not be located 1n the available
literature as dted 1n Appendix A. Chloroanlllnes are used as Intermediates
for azo and azoic dyes and pigments, pesticides, rubber chemicals (2-chloro-
anlllne), Pharmaceuticals (3- and 4-chloroan1l1ne) and agricultural chemi-
cals {KouMs and Northcott, 1963; Hawley, 1981; Kuney, 1985). 2-Chloro-
anHlne 1s also used as a standard for colorlmetrlc apparatus (Hawley, 1981).
Chloroanlllnes are weak bases and as such, may be protonated under
acidic conditions and form salts that are much more water soluble than the
parent compound. Protonatlon may cause variations 1n the behavior of
Chloroanlllnes. If released to the atmosphere, these compounds are expected
to exist almost entirely 1n the vapor phase (E1senre1ch et al., 1981).
These compounds should be removed from the atmosphere primarily by reaction
with photochemlcally generated hydroxyl radicals (estimated half-lives of 3
hours to 2 days) (Atkinson, 1985; U.S. EPA, 1987b) and possibly by direct
photolysis (FreHag et al., 1985). If released to water, chloroanlUnes are
expected to photooxldlze on the water surface (half-lives estimated at 0.5
1v
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hours) (Zepp and Schlotzhauer, 1983) and undergo rapid chemical binding with
humlc materials and clay 1n the water column and In the sediment (Parrls,
1980). Small amounts of these compounds may be removed by volatilization or
aerobic blodegradatlon by acclimated microorganisms (Kllzer et a!., 1979;
El-Dlb and Aly, 1976). The half-life of 4-chloroanlllne has been estimated
to range from 0.3-3 days In rivers and 30-300 days 1n groundwaters (Zoeteman
et al., 1980). If released to soil, chloroanlllnes will undergo rapid
chemical binding with soil components (Bollag et al., 1978; Frletag et al.,
1984) and be partially removed by chemical and biological activity (Bollag
et al., 1978; Furukawa and Brlndley, 1973; Cloos et al., 1979). Volatili-
zation from soil surface shoulxl account for a loss of only a few percent
(Fletcher and Kaufman, 1980).
Chloroanlllnes may be released to the environment as fugitive emissions
or 1n wastewater during their production or use as a chemical Intermediate.
Chloroanlllnes may also form 1n the environment as degradation products of
various pesticides (Bollag et al., 1983; Freltag et al., 1984). The most
probable routes of human exposure are Inhalation and dermal contact 1n
occupational settings. 2-Chloroan1l1ne has been Identified In drinking
water from Cincinnati, OH, and Seattle, WA (Lucas, 1984). 2-, 3- and
4-Chloroan1l1ne have been Identified 1n drinking water and rivers 1n Germany
(Kussmaul. 1978; Kool et al., 1982). Chloroanlllnes (Isomers not specified)
have been detected 1n the effluent from a publicly owned treatment works
(Ellis et al., 1982) and 2-chloroan1line has been detected In fish collected
from a river near a sewage treatment plant (Parrls et al., 1980). 2- and
4-Chloroan1l1ne have been Identified as volatile flavor components of baked
potatoes (Coleman et al., 1981).
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There were three studies In which the toxlclty of all three chloro-
anHlnes were compared In the same test system, but these studies produced
conflicting results. Regarding 14-day LC5Qs for gupples, 2-chloroan1Hne
was most toxic, followed by 3-chloroan1l1ne; 4-chloroan1l1ne was least toxic
(Hermens et al., 1985). Yoshloka et al. (1985), however, found that
4-chloroan1l1ne was most toxic 1n Inhibiting growth of the protozoan Tetra-
hymena pyrlformls. followed by 3-chloroan1l1ne and 2-chloroan1l1ne.
Devlllers et al. (1986) found that 4-chloroan1l1ne was most potent 1n
Inhibiting luminescence of Photobacterlum phosphoreum. followed by 3- and
2-chloroan1l1ne. The lowest reported acutely toxic concentrations were 2
mg/l 4-chloroan1l1ne, a 96-hour LC5Q for bluegllls (Julln and Sanders,
1978); 1.35 mg/l 4-chloroan1l1ne, a 24-hour EC for Daphnla carlnata
(Hattorl et al., 1984); and 0.4 mg/l 4-chloroan1l1ne, a 96-hour EC1Q for
Scenedesmus subsplcatus (Geyer et al., 1985). A 14-day study by Hattorl et
al. (1984) Indicated that reproduction of Daphnla carlnata was affected by
0.0427 mg/l 4-chloroan1l1ne but not by 0.0135 mg/l.
3-Chloroan1l1ne appears to be readily absorbed orally by rats; up to 75%
of a single gavage dose was excreted 1n the urine predominately as metabo-
lites within 24 hours (Boehme and Crunow, 1969); hydroxylatlon of the
benzene ring was the primary metabolic alteration. Jjj. vitro studies Indi-
cate that N-ox1dat1on 1s the primary metabolic route for 4-chloroan1l1ne.
Metabolism studies of 2-chloroan1l1ne were not located, but N-ox1dat1on Is
expected to be the primary mechanism, as 1s the case with 4-chloroanlllne.
Pertinent data regarding the effects of Inhalation exposure to the
chloroanlUnes could not be located 1n the available literature as cited 1n
Appendix A. Effects of oral administration of chloroanlUnes are limited to
v1
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4-chloroannine. In a subchronlc dietary study of 4-chloroan1l1ne, rats
treated at >680 ppm had enlarged spleens with plaque formation; rats treated
at <380 ppm did not (NCI, 1979). Enlarged spleens also occurred In mice fed
at dietary levels >11,830 ppm. At 8080 ppm, all the mice died of unknown
causes. No effects were observed at 5500 ppm.
Based on an acute oral study using cats, 4-chloroan1l1ne was -4 times
more potent as a methemogloblnemla Inducer than were 2- or 3-chloroan1l1ne
(McLean et al., 1969).
4-Chloroan1l1ne was administered 1n the diet to rats at concentrations
of 250 or 500 ppm and mice at concentrations of 2500 and 5000 ppm for 78
weeks, followed by observation periods of 24 and 13 weeks, respectively
(NCI, 1979). Effects 1n the rats Included reduced survival In the high-dose
males, reduced body weight gain In the high-dose females, flbrosls of the
splenic capsule with subcapsular mesenchymal proliferation In most of the
treated males and females, and Increased Incidences of splenic flbromas and
sarcomas 1n the high-dose males. Effects In the mice Included markedly
reduced body weight gain 1n the high- and low-dose males and females, 1ntra-
cellular deposition of hemoslderln 1n many tissues In most of the treated
males and females and hemanglomatous tumors 1n the treated males and
females. NCI (1979) concluded that these findings were suggestive of the
cardnogenldty of 4-chloroan1l1ne 1n rats and mice. The splenic tumors In
the rats were considered strongly suggestive of cardnogenldty, however,
because of the rarity of these tumors 1n historical controls. Furthermore,
preliminary results of an NTP gavage study using rats and mice Indicate an
Induction of splenic tumors 1n male rats (Canter, 1985).
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Specific Information regarding the cardnogenlcUy of 2- or 3-chloroanl-
Une could not be located In the available literature as cited In Appendix A.
Evidence for the N-ox1dat1on of monochloroanlUnes (see Section 5.3) and
methemoglobln Induction by monochloroanlllnes (McLean et al., 1969) provides
an Indication of potential carclnogenlcHy, however, as Induction of both
carclnogenlcHy and methemoblob1nem1a by aniline and substituted aniline
compounds 1s attributed to the formation of N-ox1d1zed metabolites (U.S.
EPA, 1984).
Genotoxoclty and cell transformation testing of 2-, 3- and 4-chloro-
anlllne produced positive responses 1n several assays. Conflicting results,
a lack of corroborating data and, In the case of 2- and 3-chloroan1l1ne, a
*
limited variety of assay types Indicate that evidence for genotoxiclty of
the chloroanlllnes should be regarded as Inconclusive.
Specific Information regarding the teratogenldty or other reproductive
effects of the monochloroanlUnes could not be located 1n the available
literature as dted 1n Appendix A; however, the potential for these effects
has been suggested by the U.S. EPA (1984) on the basis of possible anoxia
resulting from methemogloblnemla and reproductive effects produced by other
aniline compounds.
Given the strortg suggestion of carclnogenlcHy of 4-chloroanlllne based
on the Induction of rare splenic tumors In male rats and hemanglosarcomas 1n
female and male mice (MCI, 1979), and because preliminary results of an NTP
study Indicate that gavage treatment of rats also resulted In splenic tumors
(Canter, 1985), the evidence was considered provisionally limited and
4-chloroan1l1ne was placed 1n EPA Group C (although some may consider It
B2), a possible human carcinogen. A reanalysls of the classification should
be conducted after the new NTP bloassay Is completed In 1988. A q of
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3.5x10~2 (mg/kg/day)"1 for oral exposure was derived. The concentra-
tions In drinking water associated with Increased lifetime risk of cancer at
risk levels of 10~5, 10~» and 10~7 are IxlO'2, IxlCT3 and lx!0~*
mg/i, respectively. An F factor of 4.1xlO~1 (mg/kg/day)"1 was calcu-
lated, placing 4-chloroanlllne In Potency Group 3. A Potency Group 3 and
EPA Group C chemical has a low hazard ranking; therefore, the RQ based on
cardnogenldty 1s 100. An RQ based on chronic tox1c1ty of 1000 was also
derived based on reduced survival of rats (NCI, 1979).
The derivation of a provisional RfD of 0.01 mg/kg/day or 0.9 mg/day for
a 70 kg human, which has been proposed but not verified (U.S. EPA, 1986c),
was also presented. The RfD was based on the LOAEL of 12.5 mg/kg/day (250
ppm) at which rats had proUferatlve lesions of the spleen (NCI, 1979). An
uncertainty factor of 1000 was used. Low confidence was placed In this RfD
because of the rare splenic neoplasms also observed 1n rats In this study.
The final results of the NTP study may provide more definitive evidence, as
preliminary results suggest that rats treated by gavage developed spleen
sarcomas (Canter, 1985). It Is recommended that the provisional q,*s,
rather than the RfD, be used for regulatory purposes until the NTP report Is
available.
Data were Insufficient to derive any risk assessment or RQ values for
2- and 3-chloroan1l1ne.
1x
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TABLE OF CONTENTS
1. INTRODUCTION 1
1.1. STRUCTURE AND CAS REGISTRY NUMBER 1
1.2. PHYSICAL AND CHEMICAL PROPERTIES 2
1.3. PRODUCTION DATA 2
1.4. USE DATA 5
1.5. SUMMARY 5
2. ENVIRONMENTAL FATE AND TRANSPORT 6
2.1. AIR 6
2.1.1. Reaction with Hydroxyl Radicals 6
2.1.2. Reaction with Ozone 6
2.1.3. Photolysis 6
2.1.4. Physical Removal Processes 7
2.2. WATER ^ 7
2.2.1. Hydrolysis 7
2.2.2. Photoox1dat1on 7
2.2.3. M1crob1al Degradation 7
2.2.4. B1oconcentrat1on 8
2.2.5. Adsorption 10
2.2.6. Volatilization ' 10
2.2.7. Persistence 10
2.3. SOIL 11
2.3.1. Chemical Degradation 11
2.3.2. M1crob1al Degradation 11
2.3.3. Adsorption 12
2.3.4. Volatilization 13
2.3.5. Persistence 13
2.4. SUMMARY 14
3. EXPOSURE 15
3.1. WATER 15
3.2. FOOD 16
3.3. INHALATION 16
3.4. DERMAL 16
3.5. SUMMARY 16
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TABLE OF CONTENTS (cont.)
Page
4. AQUATIC TOXICITY 18
4.1. ACUTE TOXICITY 18
4.2. CHRONIC EFFECTS 22
4.3. PLANT EFFECTS 22
4.4. SUMHARY 22
5. PHARMACOKINETCS 26
5.1. ABSORPTION 26
5.2. DISTRIBUTION 26
5.3. METABOLISM 26
5.4. EXCRETION 27
5.5. SUMMARY 28
6. EFFECTS 29
6.1. SYSTEMIC TOXICITY. '..... 29
6.1.1. Inhalation Exposures 29
6.1.2. Oral Exposures 29
6.1.3. Other Relevant Information 31
6.2. CARCINOGENICITY 32
6.2.1. Inhalation '. 32
6.2.2. Oral 32
6.2.3. Other Relevant Information 37
6.3. MUTAGENICITY 37
6.4. TERATOGENICITY 45
6.5. OTHER REPRODUCTIVE EFFECTS 45
6.6. SUMMARY 45
7. EXISTING GUIDELINES AND STANDARDS 48
7.1. HUMAN 48
7.2. AQUATIC 48
8. RISK ASSESSMENT 49
8.1. CARCINOGENICITY 49
8.1.1. Inhalation 49
8.1.2. Oral 49
8.1.3. Other Routes 51
8.1.4. Weight of Evidence 51
8.1.5. Quantitative Risk Estimates 51
x1
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TABLE OF CONTENTS (cont.)
Page
8.2. SYSTEMIC TOXICITY 53
8.2.1. Inhalation Exposure 53
8.2.2. Oral Exposure 53
9. REPORTABLE QUANTITIES 55
9.1. BASED ON SYSTEMIC TOXICITY 55
9.2. BASED ON CARCINOGENICITY 57
10. REFERENCES 63
APPENDIX A: LITERATURE SEARCHED 85
APPENDIX B: CANCER DATA SHEET FOR DERIVATION OF q-.*s 88
APPENDIX C: SUMMARY TABLE FOR 4-CHLOROANILINE 90
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No.
1-1
1-2
4-1
4-2
4-3
6-1
6-2
6-3
6-4
9-1
9-2
9-3
9-4
9-5
LIST OF TABLES
Title
Selected Physical Properties of ChloroanlUnes
Current Manufacturers of ChloroanHlnes
Acute Toxlclty of ChloroanlUnes to Freshwater Fishes ....
Acute Toxlclty of ChloroanlUnes to Invertebrates
Toxlclty of ChloroanlUnes to Freshwater Plants and
Bacteria
Incidences of Tumors 1n F344 Rats and B6C3F1 Mice Treated
with Technical Grade 4-Chloroan1l1ne 1n the Diet
MutagenlcHy and Other Short-Term Assays of
2-Chloroan1l1ne
MutagenlcHy and Other Short-Term Assays,of
3-Chloroan1l1ne *
MutagenlcHy and Other Short-Term Assays of
4-Chloroan1l1ne
Oral Toxlclty Summary for 4-Chloroan1l1ne (technical
grade) 1n Diet
Oral Composite Scores for 4-Chloroan1l1ne Using the Rat . . .
4-Chloroan1l1ne: Minimum Effective Dose (MED) and
Reportable Quantity (RQ)
2- and 3-Chloroan1l1ne: Minimum Effective Dose (MEO) and
Reportable Quantity (RQ)
Derivation of Potency Factor (F) for 4-Chloroan1l1ne
Page
3
4
19
21
23
34
38
40
41
56
58
59
60
61
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LIST OF ABBREVIATIONS
BCF Bloconcentratlon factor
BUN Blood urea nitrogen
CS Composite score
ONA Deoxyrlbonuclelc acid
ECso Concentration effective to 50% of recipients
Koc Soil sorptlon coefficient standardized
with respect to soil organic matter
Kow Octanol/water partition coefficient
LCg Concentration lethal to 0% of recipients
LCso Concentration lethal to 50% of recipients
LOAEL Lowest-observed-adverse-effect level
MED . Minimum effective dose
NOAEL No-observed-adverse-effect level
NOEC No-observed-effect concentration
ppb Parts per billion
ppm Parts per million
RfO Reference dose
RQ Reportable quantity
RV,j Dose-rating value
RVe Effect-rating value
UV Ultraviolet
x1v
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1. INTRODUCTION
1.1. STRUCTURE AND CAS REGISTRY NUMBER
The synonyms, structure and CAS Registry number for 2-, 3- and 4-chloro-
an111ne are as follows:
2-Chloroanlllne
CAS Registry number: 95-51-2
Synonyms: ortho-Chloroan1l1ne; l-am1no-2-chlorobenzene; 2-chlorophenyl-
amlne; 2-chlorobenzenam1ne; Azoic Dlazo Component 44; CI 37000 (KouMs and
Northcott, 1963); Fast Yellow GC Base (U.S. EPA, 1987a):
Structure:
3-Chloroan1l1ne
CAS Registry number: 108-42-9
Synonyms: meta-Chloroan111ne; l-amlno-3-chlorobenzene; 3-chlorophenylam1ne;
3-chlorobenzenam1ne; Azoic Dlazo Component 2; CI 37005 (Kourls and
Northcott. 1963); Fast Orange GC Base (U.S. EPA. 1987a)
Structure:
4-Chloroan1l1ne
CAS Registry number: 106-47-8
Synonyms: para-chloroanlllne; l-am1no-4-chlorobenzene; 4-chloropheny1am1ne;
4-ch1orobenzenam1ne (U.S. EPA, 1987a).
0028d
-1-
05/15/87
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Structure:
Cl
Each of the chloroanlUne Isomers has a molecular weight of 127.58 and the
empirical formula C^H^CIM.
1.2. PHYSICAL AND CHEMICAL PROPERTIES
2-Chloroan1l1ne 1s an amber liquid; 3-chloroan1l1ne Is a colorless to
light amber liquid; and 4-chloroan111ne 1s a white to pale yellow solid at
room temperature (Hawley, 1981). Aromatic amines are oxidized easily by air
^
and chloroanllines have a tendency to darken on exposure to air because of
the formation of oxidation products (Hawley. 1981; Morrison and Boyd, 1973).
Aromatic amines are weak bases and may be readily converted to their salts
by aqueous mineral adds or carboxyllc adds. Aqueous hydroxyllons can
readily convert these salts back to the free amlne (Morrison and Boyd,
1973). Chloroanlllnes are soluble 1n most common organic solvents {Hawley,
1981; Hlndholz, 1983). Selected physical properties for the ch1oroan1!1ne
Isomers are listed In Table 1-1.
1.3. PRODUCTION DATA
Halogenated anilines such as chloroanlUnes are usually produced by
reduction of the corresponding nltro compounds 1n the presence of Iron and
hydrochloric add. Chloroanlllnes may also be made by the ammonolysls of
the appropriate bromochlorobenzene (Kourls and Northcott, 1963). Data
regarding current domestic manfacturers of Chloroanlllnes are listed In
Table 1-2. Domestic production volume data could not be located 1n the
available literature as cited In Appendix A. The most recent year for which
0028d -2- 06/05/87
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0028d
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-------
TABLE 1-2
Current Manufacturers of ChloroanlUnes*
Chemical
Manufacturer
Location
2-Chloroan1l1ne
3-Chloroan111ne
4-Chloroan1l1ne
E.I. Dupont
The Upjohn Co.
E.I. Dupont
First Mississippi Corp.
E.I. Oupont
Monsanto Co.
Deepwater, NJ
North Haven, CT
Oeepwater, NJ
Pascagoula, MI
Deepwater, NJ
Lullng, LA
*Source: SRI, 1986
0028d
-4-
05/15/87
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Import data are available 1s 1983, during which 179,649 pounds of 2-chloro-
anlUne and 830,915 pounds of 3-chloroan1l1ne were Imported through the
principle U.S. customs districts (USITC, 1984).
1.4. USE DATA
ChloroanlUnes are used as Intermediates for azo and azoic dyes and
pigments, pesticides, rubber chemicals (2-chloroan1l1ne), Pharmaceuticals
(3- and 4-chloroan1l1ne) and agricultural chemicals (Society of Dyers and
Colorlsts, 1971; Kourls and Northcott, 1963; Hawley, 1981; Kuney, 1985).
2-Chloroan1l1ne 1s also used as a standard for colorlmetMc apparatus
(Hawley, 1981).
1.5. SUMMARY
*»
2-Chloroan1l1ne (CAS number 95-51-2) 1s an amber liquid; 3-chloroan1l1ne
(CAS number 108-42-9) 1s a colorless to light amber liquid; and 4-chloro-
anlUne (CAS number 106-47-8) 1s a white to pale yellow solid at room
temperature (Hawley, 1981). They are slightly soluble 1n water and are
soluble 1n common organic solvents (Hawley, 1981). The 1986 Directory of
Chemical Producers (SRI, 1986) reports that E.I. Dupont 1n Deepwater, NJ,
currently produces all three chloroanHlne Isomers; The Upjohn Co. 1n North
Haven, CT, produces 2-chloroan1l1ne; First Mississippi Corp. produces
3-chloroan1l1ne; and Monsanto Co. In Lullng, LA, produces 4-chloroan1l1ne.
Domestic production volume data for recent years could not be located 1n the
available literature as cited In Appendix A. ChloroanlUnes are used as
Intermediates for azo and azoic dyes and pigments, pesticides, rubber
chemicals (2-chloroan1l1ne), Pharmaceuticals (3- and 4-chloroan1l1ne) and
agricultural chemicals (Kourls and Northcott, 1963; Hawley, 1981; Kuney,
1985). 2-Chloroan1l1ne 1s also used as a standard for colorlmetrlc
apparatus (Hawley, 1981).
0028d -5- 06/05/87
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2. ENVIRONMENTAL FATE AND TRANSPORT
ChloroanHlnes are weak bases [pKa 2.66-3.98 (PerMn, 1972)] which may
be protonated under addle conditions to form salts that are much more water
soluble than the parent compound. Protonatlon can alter the behavior of a
compound In water and soil, for example, by decreasing volatilization as
well as decreasing or Increasing physical adsorption to soil, sediments and
suspended solids 1n water.
2.1. AIR
Based on the vapor pressures listed In Table 1-1, the chloroanlllnes are
expected to exist almost entirely 1n the vapor phase 1n the atmosphere
(Elsenrelch et al., 1981).
2.1.1. Reaction with Hydroxyl Radicals. Using experimentally determined
hydroxyl reaction rate constants of 8.30x10'" cmVmolecule-sec at 22°C
(Atkinson, 1985) and 8.09x10"" cmVmolecule-sec at 27*C (Gusten et al.,
1984) and an ambient hydroxyl radical concentration of 8.0x10* mole-
cules/cm3 (U.S. EPA, 1987b), the half-life for 4-chloroan1l1ne vapor
reacting with photochemlcally generated hyroxyl radicals In the atmosphere
has been estimated to be ~3 hours. The half-lives for 2-, 3- and 4-chloro-
anHlne vapor reacting with photochemlcally generated hydroxyl radicals have
been estimated to be -2 days based on an estimated reaction constant of
5.1xlO~12 cmVmolecule-sec at 25°C (U.S. EPA, 1987b).
2.1.2. Reaction with Ozone. Chloroanlllnes are not susceptible to oxida-
tion by ozone 1n the atmosphere (U.S. EPA, 1987b).
2.1.3. Photolysis. 4-Chloroan1l1ne adsorbed onto silica gel underwent
27.7% mineralization when Irradiated with light of wavelengths >290 nm for
17 hours (Freltag et al., 1985). Chloroanlllnes In methanol or cyclohexane
0028d -6- 05/15/87
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strongly absorb UV light 1n the environmentally significant wavelength range
of >290 nm (Sadtler Research Laboratory, 1960a,b, 1962). These data
Indicate that chloroanlUnes have the potential to undergo direct photolysis
In the atmosphere.
2.1.4. Physical Removal Processes. Based on the water solubilities
listed In Table 1-1, It appears that small amounts of chloroanlUnes may be
removed from the atmosphere 1n wet precipitation.
2.2. WATER
2.2.1. Hydrolysis. Halogenated aromatlcs and aromatic amines are
generally resistant to hydrolysis (Lyman et al., 1982); thus, chloroanlUnes
are expected to be resistant to hydrolysis.
2.2.2. Photoox1dat1on. The sunlight-Induced" photolysis half-life of
4-chloroan1l1ne In distilled water has been estimated to be -0.5 hours using
an experimentally determined reaction rate constant of 1.76 hr"1. This
rate was not significantly Increased by the presence of algae 1n the water
(Zepp and Schlotzhauer, 1983). No transformations 1n 4-chloroan1l1ne were
observed when aqueous solutions of this compound were kept In the dark for
3-4 hours (Zepp and Schlotzhauer, 1983). Irradiation of 10"* M 4-chloro-
anlllne In air-saturated water with UV light of wavelengths >290 nm produced
4-chloronltrosobenzene and 4-chloronltrobenzene; 4-chloroan1l1ne would not
be detected after 6 hours of Irradiation (Miller and Crosby, 1983). 2- and
3-Chloroan1l1ne exposed to UV light of wavelengths >300 nm underwent 50%
degradation after 7.5 and 11.5 minutes, respectively (Kondo, 1978).
2.2.3. M1crob1al Degradation. The results of blodegradatlon screening
studies on chloroanlUnes range from no degradation or m1crob1al Inhibition
to rapid degradation using freshwater, activated sludge and sewage as
Inocula (GeMke and Fischer, 1981; Kawasaki, 1980; King and Painter. 1983;
0028d -7- 05/15/87
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Janlcke and H1lge, 1980; El-D1b and Aly, 1976; Malaney, 1960; Thorn and Agg,
1975; Balrd et al., 1977; PHter, 1976; Schmldt-Bleek et al., 1982; FreHag
et al., 1985). The most frequently reported results Indicate that chloro-
anlUnes degrade slowly with acclimation (Thorn and Agg, 1975; Balrd et al.,
1977; PUter, 1976; Schmldt-Bleek et al., 1982; Halaney, 1960; El-01b and
Aly, 1976). In one river die-away study, 10 ppm 4-chloroan1l1ne degraded
slowly 1n Nile River water over a period of 2 months; on redose, however,
Increasingly larger concentrations were degraded over a diminishing period
of time. On the eighth redose, 100 ppm 4-chloroanlllne was degraded In just
a few days (El-D1b and Aly, 1976). A proposed pathway for the blodegrada-
tlon of 2-chloroan1l1ne 1s shown 1n Figure 2-1.^ The first Intermediates 1n
the metabolism of chloroanlUnes by £_._ multlvorans strain An.l were chloro-
catechols (Reber et al., 1979). Pseudomonas aurantlaca converted 4-chloro-
anlllne to Us corresponding acetanlUde (Surovtseva et al., 1977). No
mineralization of 4-chloroan1l1ne occurred when this compound was Incubated
with digester sludge under anaerobic conditions for 1 month (Shelton and
Tledje, 1981).
2.2.4. Blconcentratlon. BCF values of 13-20 1n the In the Golden orfe,
Leudsens Idus melanotus. and 260-1200 In algae, Chlorella fusca. have been
measured for 4-chloroan1l1ne (Korte et al., 1978; FreHag et al., 1985;
Geyer et al., 1984). Based on the log K values listed In Table 1-1,
B'CFs of 16, 16 and 14 were estimated for 2-, 3- and 4-chloroan1l1ne, respec-
tively, using the following recommended linear regression equation (Lyman et
al., 1982): log BCF « 0.76 log K - 0.23. These BCF values suggest that
bloaccumulatlon of chloroanlUnes 1n aquatic organisms would not be signifi-
cant.
0028d -8- 06/05/87
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HOOC
COOH ci
COOH
COOH
HOOC
COOH
CH3COOH +
*HOOC-CH=CH-COOH
FIGURE 2-1
Proposed B1odegradat1on Pathway of 2-Ch1oroan1"Mne
Source: Rump, 1984
0028d
-9-
05/15/87
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2.2.5. Adsorption. 2-Chloroan1l1ne and 4-chloroan1l1ne have been ob-
served undergoing rapid, reversible covalent binding with humates In aqueous
solution. This reaction 1s believed to represent the formation of 1m1ne
linkages with humate carbonyl groups. This Initial binding reaction Is
followed by a much slower, less readily reversible binding reaction, which
Is believed to represent the addition of the amines to qulnoldal structures
followed by oxidation of the product to a nitrogen substituted qulnold ring
(ParMs, 1980). The model half-life of 4-chloroanHlne binding with one
test humlc constituent, p-benzoqu1none, was 13 minutes (ParMs, 1980).
Based on these observations, 1t Is expected that chemical binding of chloro-
anlUnes to humlc substances present In sedlmepts and suspended solids 1n
water would be significant.
2.2.6. Volatilization. Under laboratory conditions, 50 ppb 4-chloro-
anlUne 1n aqueous solution underwent 0.63% loss, which was due to volatili-
zation after 2 hours (Kllzer et al., 1979). Henry's Law constants calcu-
lated from vapor pressure and water solubility data listed 1n Table 1-1 are
7.5xlO~* atm-mVmol at 20°C for 3-chloroan1l1ne, 1.7xlO~* atm-mVmol
at 20°C for 3-chloroan1l1ne and 1.2xlO~* atm-mVmol at -25°C for
4-chloroan1l1ne. Based on these values, the volatilization half-life from a
typical river 1 m deep, flowing 1 m/sec with a wind speed of 3 m/sec has
been estimated to be 5.5, 24 and 34 days for 2-, 3- and 4-chloroanlllne,
respectively, using the method of Lyman et al. (1982).
2.2.7. Persistence. The half-life of 4-chloroan1l1ne has been estimated
to range from 0.3-3 days 1n rivers and from 30-300 days 1n groundwaters
(Zoeteman et al., 1980). When 4-chloroan1l1ne with trace amounts of the
14C-labeled material was applied to experimental ponds for 4-6 weeks at an
average concentration of -50 yg/l, 14C disappeared from the water 1n
0028d -10- 06/05/87
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two phases with half-lives of ~3 and 11 days, respectively. It was assumed
that Initial loss was the result of volatilization. After 3 days, products
of autooxldatlon, with lower volatility, caused a decrease 1n the rate of
14C loss from water. No significant leaching horizontally or vertically
Into the soil surrounding the experimental pond was observed. One year
after application, -10-20X of the total 14C applied was present as bound
residues 1n the upper 0- to 10-cm soil layer (Schauerte et al., 1982). In 2
weeks, 250 vq/l 3- and 4-chloroan1l1ne added to separate eutrophlc pond
water samples underwent 7 and 4X removal, respectively. In a similar study,
3- and 4-chloroan1l1ne added to eutrophlc pond water samples with sewage
sludge Inoculum underwent 9 and 7% removal, respectively, In 2 weeks (Lyons
*
et al., 1985). At 30°C, 0.1 wl/mi 3-chloroanlllne Incubated 1n three
different river water samples In the dark had a half-life of 3-7 days.
Under the same test conditions using two different samples of sea water,
this compound had a half-life of -3 days (Kondo, 1978).
2.3. SOIL
2.3.1. Chemical Degradation. Pertinent data regarding the chemical
degradation of chloroanHlnes In soil other than those given 1n Section
2.3.3. could not be located 1n the available literature as cited 1n
Appendix A.
2.3.2. M1crob1al Degradation. In autoclaved and nonautoclaved soil
treated with 5 ppm [l4C]-4-chloroanH1ne and Incubated for 6 weeks, no
14C02 evolution was observed 1n the autoclaved sample, but 7.5X
14C02 was released from the nonautoclaved soil (Bollag et al., 1978).
Alexander and Lustlgman (1966) found 10 yg/l 2-, 3- and 4-chloroan1l1ne
Inoculated with mixed cultures of soil microorganisms to be resistant to
degradation. A Pseudomonas species Isolated from soil and grown under
0028d -11- 05/15/87
-------
aerobic conditions used 4-chloroanlUne as a sole source of C and M. A mass
balance on [l*C]-4-chloroanH1ne revealed that 64% of **C was released
as CO- and 14% was associated with the blomass (Zeyer and Kearney, 1982).
Fusarlum oxysporum Schlecht Isolated from soil-degraded 4-chloroan1l1ne
through at least two metabolic pathways. Oxidation of the amlne group was
the primary mechanism, but acylatlon of the amlne group also occurred
(Kaufman et al., 1973). A Paracoccus species Isolated from soil and grown
under anaerobic conditions metabolized 4-chloroan1l1ne Into a product
Identified as 1,3-b1s(p-chlorophenyl)tr1azene (Mlnard et al., 1977).
2.3.3. Adsorption. The K of 4-chloroan1l1ne has been determined to
range from 96-1530 In a variety of soils. Lower soil pHs produced lower
K values Indicating the Importance of covalent binding to soils (Van
Blade! and Horeale, 1977; Rlppen et al., 1982). After Incubating 5 ppm
[l4C]-4-chloroan1!1ne In autoclaved and nonautoclaved soil (3.4% organic
matter) for 6 weeks, 71.0 and 72.4%, respectively, of total 14C applied
was found to exist as soil-bound residues (Bo"Mag et al., 1978). Based on
the results of a 3-year field lyslmeter study, Freltag et al. (1984)
speculated that rapid binding Immobilizes 4-chloroan1l1ne 1n soil and only
very small amounts of free chloroanlllne are able to leach Into deeper soil
layers. Bartha (1971) determined that Immobilized chloroanlUnes are
chemically bound to humlc substances and physical adsorption Is, at most, a
secondary process. Studies by other Investigators also Indicate that the
chloroanlllne should undergo rapid, tight binding with humlc substances 1n
soil (Worobey and Webster, 1982; Bollag et al., 1983; ParMs, 1980).
Adsorption and oxidation of aromatic amines can occur on clay surfaces but
1s dependent on the exchangeable cation In the clay and the presence of
oxygen (Furukawa and BMndley, 1973; Cloos et al., 1979), as Indicated by
0028d -12- 06/05/87
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the formation of olIgomeMc and polymeric complexes when 4-chloroan1l1ne 1s
adsorbed onto montmorlllonlte clay (Cloos et al., 1979). The K value
for 4-chloroan1l1ne adsorbed onto colloidal organic matter present 1n
groundwater samples was found to be 5550 (Means, 1983), which suggests that
adsorption onto this mlcropartlculate matter could effectively Increase the
solubility and leaching of chloroanlUnes Into landfill groundwater.
2.3.4. Volatilization. Under laboratory conditions, 4-chloroan1l1ne
applied to sand, loam and humus underwent 2.01, 0.72 and 0.11% loss, respec-
tively, 1n 2 hours, which was due to volatilization (KHzer et al., 1979).
After 49 days, <2% of 3-chloroan1l1ne applied to unsteMUzed Hagerstown
sllty clay loam was found to have volatilized (Hetcher and Kaufman, 1980).
Based on the strong binding of chloroanlllnes to soil organic matter and the
partial protonatlon of these compounds In acidic soils (see pKa values
listed 1n Table 1-1), volatilization from wet and dry soil surfaces 1s not
expected to be significant.
2.3.5. Persistence. When 50 mg [l4C]-4-chloroan1Hne, which corre-
sponded to 1.25 ppm 1n soil to a depth of 10 cm, was added to a field
lyslmeter, a total of 32.8% of the 14C applied was recovered 20 weeks
later: 32.4% 1n soil, 0.3% 1n plants and 0.1% In leached water. Of the
total 4-chloroan1l1ne present 1n the soil, >90% was 1n the form of soil-
bound residues (Freltag et al., 1984). A 16-week study of 2-, 3- and
4-chloroan1l1ne 1n two different soils revealed that 3-chloroanlHne
degradation (10.6-11.9%) was the fastest of the three Isomers (Fuchsblchler
et al., 1978). During a 16-week study of this compound 1n four agricultural
soils, 12.3-17.2% mineralization of 4-chloroan1l1ne was observed (Suess et
al., 1978).
0028d -13- 06/05/87
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2.4. SUMMARY
Chloroan111nes are weak bases and as such, may be protonated under
acidic conditions and form salts that are much more water soluble than the
parent compound. Protonatlon may cause variations In the behavior of
chloroanlUnes. If released to the atmosphere, these compounds are expected
to exist almost entirely 1n the vapor phase (E1senre1ch et al., 1981).
These compounds should be removed from the atmosphere primarily by reaction
with photochemically generated hydroxyl radicals (estimated half-lives of 3
hours to 2 days) (Atkinson, 1985; U.S. EPA. 1987b) and possibly by direct
photolysis (FreHag et al., 1985). If released to water, chloroanlUnes are
expected to photooxldlze on the water surface (-half-lives estimated at 0.5
hours) (Zepp and Schlotzhauer, 1983) and undergo rapid chemical binding with
humlc materials and clay 1n the water column and 1n the sediment (Parrls,
19.80). Small amounts of these compounds may be removed by volatilization or
aerobic blodegradatlon by acclimated microorganisms (Kllzer et al., 1979;
El-01b and Aly, 1976). The half-life of 4-chloroan1l1ne has been estimated
to range from 0.3-3 days 1n rivers and 30-300 days In groundwaters (Zoeteman
et al., 1980). If released to soil, chloroanlUnes will undergo rapid
chemical binding with soil components (Bollag et al., 1978; Frletag et al.,
1984) and be partially removed by chemical and biological activity (Bollag
et al., 1978; Furukawa and BMndley, 1973; Cloos et al., 1979). Volatili-
zation from soil surface should account for a loss of only a few percent
(Fletcher and Kaufman, 1980).
0028d -14- 06/05/87
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3. EXPOSURE
The National Occupational Hazard Survey (NOHS) prepared by NIOSH
estimates that 18,138 workers are occupatlonally exposed to 2-chloroan1l1ne
(NIOSH, 1984). NOHS estimates were not provided for the other chloroanHlne
Isomers. The most probable routes of human exposure are Inhalation and
dermal contact In occupational settings.
Chloroanlllnes may be released to the environment as fugitive emissions
or 1n wastewater during their production or use as chemical Intermediates.
Chloroanlllnes may also form 1n the environment as degradation products of
various pesticides (Bollag et al., 1983; Fre1tag,et a!., 1984; Suess, 1973).
3.1. WATER
2-Chloroan1l1ne has been positively Identified 1n drinking water samples
obtained from Cincinnati, OH, In 1980 and Seattle, HA, 1n 1976 (Lucas,
1984). The mean concentration of 4-chloroan1l1ne In German drinking water
(treated Rhine water) from September 1973 to September 1974 was 7 ng/l
(Kussmaul, 1978). 2- and 3-Chloroan1l1ne have also been detected In
drinking water In Germany (Kool et al., 1982).
During 1979, the mean concentration of 2-, 3- and 4-chloroan1l1ne 1n the
Rhine River at LobUh was 0.52, 0.14 and 0.22 yg/l, respectively (Wegman
and Oekorte, 1981). During 1979, the mean concentration of 2-, 3- and
4-chloroan1l1ne 1n the Meuse River at Eljsden was 0.86, 0.06 and 0.08
jig/1, respectively, and at L1th, 0.06, 0.02 and 0.01 jig/l, respec-
tively (Wegman and Dekorte, 1981). 4-Chloroan1l1ne was detected 1n the
Rhine River from September 1973 to September 1974 at a mean concentration of
100 ng/l (Kussmaul, 1978). 2-, 3- and 4-Chloroan1l1ne have also been
found 1n the delta waters from the Rhine River (Greve and Wegman, 1975).
0028d -15- 06/05/87
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Chloroanlllnes (Isomers not specified) have been detected In the
effluent from the publicly owned treatment works (POTW) of Sauget, IL (Ellis
et al., 1982). In Adrian, MI, 600 ppm (dry weight basis) 2-chloroan1l1ne
was found 1n the sediment of a waste treatment lagoon of a small
4,4'-methyleneb1s(2-chloroan1l1ne) manufacturer. This compound has also
been detected In fish collected from a river near the Adrian, MI, sewage
treatment plant (Parrls et al., 1980).
Since 4-chloroan1l1ne readily adsorbs onto colloidal organic matter and
adsorption onto m1cropart1culates> could effectively Increase the
leachabllHy of the chloroanlllnes from landfills Into groundwater, ground-
water 1s theoretically a potentially significant source of human exposure.
3.2. FOOD
2- and 4-Chloroan1l1ne have been Identified as volatile flavor compo-
nents of Idaho Russet Burbank baked potatoes (Coleman et al., 1981).
3.3. INHALATION
Pertinent data regarding exposure by Inhalation could not be located In
the available literature as cited 1n Appendix A.
3.4. DERMAL
Pertinent data regarding exposure by dermal contact could not be located
In the available literature as cited in Appendix A.
3.5. SUMMARY
Chloroanlllnes may be released to the environment as fugitive emissions
or 1n wastewater during their production or use as a chemical Intermediate.
Chloroanlllnes may also form 1n the environment as degradation products of
various pesticides (Bollag et al., 1983; Freltag et al., 1984). The most
probable routes of human exposure are Inhalation and dermal contact In
occupational settings. 2-Chloroan1l1ne has been Identified 1n drinking
0028d -16- 07/21/87
-------
water from Cincinnati, OH, and Seattle, HA (Lucas, 1984). 2-, 3- and
4-Ch1oroan1l1ne have been Identified In German drinking water and German
rivers (Kussmaul, 1978; Kool et al., 1982). ChloroanHlnes (Isomers not
specified) have been detected In the effluent from a publicly owned treat-
ment works (Ellis et al., 1982) and 2-chloroan111ne has been detected In
fish collected from a river near a sewage treatment plant (Parrls et al.,
1980). 2- and 4-Chloroan111ne have been Identified as volatile flavor
components of baked potatoes (Coleman et al., 1981).
0028d -17- 07/21/87
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4. AQUATIC TOXICITY
4.1. ACUTE
The available data concerning acute toxldty of chloroanlllnes to fresh-
water fishes are presented 1n Table 4-1. The only study 1n which all three
chloroanlllnes were tested was that of Hermens et al. (1985) 1n which 14-day
LC5Q values for gupples, Poec111a retlculata. were determined. These data
suggested that 2-chloroan1l1ne was the most toxic (LC5Q = 6.25 mg/l),
3-chloroan1l1ne was Intermediate (LC5Q = 13.36 mg/l) and 4-chloroanlllne
was the least toxic {LC50 - 26.05 mg/l). The most sensitive species
tested appeared to be the blueglll, Lepomls macrochlrus. with a 96-hour
LC5Q of 2 mg/l 4-chloroan1l1ne (Julln and Sanders, 1978), the lowest
reported acutely toxic concentration for freshwater fishes. Data regarding
marine fish species could not be located 1n the available literature as
cited In Appendix A.
ChloroanHlne acute toxldty data for Invertebrates are presented In
Table 4-2. The only one of these studies 1n which all three chloroanlllnes
were compared was that of Yoshloka et al. (1985). These Investigators found
that 4-chloroan1l1ne was much more toxic than 3-chloroan1l1ne or 2-chloro-
anlUne to the dilate protozoan, Tetrahymena pyrlformls. EC,Q values for
growth Inhibition 1n this study were 10 mg/l for 4-chloroanlllne, 100
mg/l for 3-chloroan1l1ne and 200 mg/l for 2-chloroan1l1ne. The lowest
reported toxic concentration for freshwater Invertebrates was 1.35 mg/l
4-chloroan1l1ne, a 24-hour EC5Q for the cladoceran Daphnla carlnata
(Hattorl et al., 1984). For saltwater Invertebrates, the lowest reported
toxic concentration was 10 mg/l 4-chloroan1l1ne which Impaired development
of oyster, Crassostrea v1rq1n1ca embryos (EG&G, 1979).
0028d -18- 05/15/87
-------
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4.2. CHRONIC EFFECTS
The only chronic toxldty data for chloroanlUnes were provided by
HattoM et al. (1984) who reported that reproduction of Daphnla caMnata was
affected by 4-chloroanlllne concentrations as low as 0.0427 mg/l 1n 14-day
exposures. Only the abstract of this Japanese paper was translated, and no
other details were provided. A figure In the paper seemed to Indicate that
reproduction was not significantly affected by 4-chloroanlllne at a concen-
tration of 0.0135 mg/l.
4.3. PLANT EFFECTS
The available Information concerning effects of chloroanlUnes on
aquatic plants and bacteria Is presented 1n Tabje 4-3. The lowest reported
toxic concentration was 0.4 mg/l 4-chloroan1l1ne, which was a 96-hour
EC,Q for growth Inhibition 1n the alga Scenedesmus subsplcatus (Geyer et
al., 1985). Deylllers et al. (1986) compared toxldty of the three chloro-
anlUnes with PhotobacteMum phosphoreum and found that 4-chloroan1l1ne was
most toxic, followed by 3-chloroan1l1ne and 2-chloroan1l1ne.
4.4. SUMMARY
There were three studies 1n which the toxldty of all three chloroanl-
Unes were compared 1n the same test system, but these studies produced
conflicting results. Regarding 14-day LC5Qs for gupples, 2-chloroan1l1ne
was most toxic, followed by 3-chloroan1l1ne; 4-chloroan1l1ne was least toxic
(Hermens et al., 1985). Yoshloka et al. (1985), however, found that
4-chloroan1l1ne was most toxic 1n Inhibiting growth of the protozoan
Tetrahymena pyr1form1s. followed by 3-chloroan1l1ne and 2-chloroan1l1ne.
Q028d -22- 06/05/87
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Devlllers et al. (1986) found that 4-chloroan1l1ne was most potent 1n
Inhibiting luminescence of PhotobacteMum phosphoreum. followed by 3-chloro-
anlllne and 2-chloroan1l1ne. The lowest reported acutely toxic concentra-
tions were: 2 mg/l 4-chloroan1l1ne, a 96-hour LC5Q for bluegllls (Julln
and Sanders, 1978); 1.35 mg/l 4-chloroan1l1ne, a 24-hour EC5_ for
Daphnla carlnata (Hattorl et al., 1984); and 0.4 mg/l 4-chloroanlllne, a
96-hour EC,Q for Scenedesmus subsplcatus (Geyer et al., 1985). A 14-day
study by Hattorl et al. (1984) Indicated that reproduction of Daphnla
carlnata was affected by 0.0427 mg/l 4-chloroan1l1ne but not by
0.0135 mg/l.
0028d -25- 05/15/87
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5. PHARMACOKINETICS
5.1. ABSORPTION
In a metabolism study of 3-chloroan1l1ne with rats, -75% of a 10 mg/kg
oral dose and 56% of a 150 mg/kg oral dose were excreted 1n the urine as
metabolites or unchanged compound within 24 hours (Boehme and Crunow, 1969)
(Section 5.3.), which Indicates that at least 56-75% was absorbed by the
gastrointestinal tract.
Specific data regarding the oral or Inhalation absorption of 2- or
4-chloroan1l1ne could not be located 1n the available literature as cited In
Appendix A. Oral absorption of these Isomers 1s Indicated, however, by the
occurrence of systemic toxldty following oral exposure (Chapter 6).
5.2. DISTRIBUTION
Pertinent data regarding the distribution of absorbed 2-, 3- or
4-chloroan1l1ne could not be located In the available literature as cited 1n
* *
Appendix A.
5.3. HETABOLISM
An In vivo metabolism study was conducted 1n which single 10, 60 or 150
mg/kg doses of 3-ch1oroan111ne 1n Erdnussol (vehicle) were administered by
gavage to male albino rats (Boehme and Crunow, 1969). Urinary analyses
conducted 24 hours after dosing Indicated that hydroxylatlon of the benzene
ring was the primary metabolic alteration, as 2-am1no-4-chloropheno1 and
4-am1no-2-chlorophenol and their glucuronlc or sulfurlc add conjugates
accounted for -16-25% and 39-50% of the administered doses, respectively.
Unchanged 3-chloroan1l1ne accounted for 0.6-1.2% of the administered doses.
Partial acetylatlon of the amlno groups 1n the phenolic compounds also
occurred but was not quantified.
0028d -26- 06/05/87
-------
Klese (1963) detected 3- and 4-chloronHrosobenzene In the blood of dogs
after single 25-100 mg/kg Intravenous Injections of 3- and 4-chloroan1Hne,
respectively. Also, rabbit hemoglobin-mediated N-ox1dat1on of 4-chloro-
anlUne has been demonstrated In vitro (Golly and Hlavlca, 1983).
N-hydroxylatlon of 4-chloroan1l1ne was demonstrated 1n J[n vitro studies
with rat liver mlcrosomes (Uehleke, 1967; Uehleke et al., 1971; Pan et al.,
1979; Lenk and Sterzl, 1981) and ram seminal vesicle mlcrosomes (Golly and
Hlavlca, 1985). ln_ vitro metabolism of 4-chloroan1l1ne with rabbit liver
mlcrosomes resulted 1n N-hydroxylat1on and dechlorlnatlon, producing a trace
amount of 4-hydroxyan1l1ne at a high substrate concentration (Daly et al.,
1968) and hydroxylatlon of the number 2 carbon atoms (Lenk and Sterzl, 1981).
Although metabolism studies of 2-chloroan1l1ne could not be located 1n
the available literature, N-ox1dat1on 1s expected to be the primary mecha-
nism, as 1s the case with 4-chloroan1l1ne, because chlorine 1s an ortho-,
para-d1rect1ng substUuent. Likewise, the 2- and 4-chloro Isomers are
expected to have lower electron ring densities than the 3-chloro Isomer,
which makes electrophlUc aromatic (ring) substitution less likely to occur
1n the 2- and 4-chloro Isomers (Morrison and Boyd, 1966). This 1s consis-
tent with the above data Indicating N-ox1dat1on of 4-chloroan1l1ne and ring
hydroxylatlon of 3-chloroan1l1ne as primary metabolic pathways.
5.4. EXCRETION
The rat metabolism study summarized 1n Section 5.3. (Boehme and Crunow,
1969) Indicates that substantial portions of single oral doses of 3-chloro-
anHlne are excreted In the urine as metabolites within 24 hours.
0028d -27- 05/15/87
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5.5. SUMMARY
3-Chloroan1l1ne appears to be readily absorbed orally by rats; up to 75%
of a single gavage dose was excreted 1n the urine predominately as metabo-
lites within 24 hours (Boehme and Crunow, 1969); hydroxylatlon of the
benzene ring was the primary metabolic alteration. Iji vitro studies Indi-
cate that N-ox1dat1on 1s the primary metabolic route for 4-chloroan1l1ne.
Metabolism studies of 2-chloroan1l1ne were not located, but N-ox1dat1on 1s
expected to be the primary mechanism, as Is the case with 4-chloroan1l1ne.
0028d -28- 06/05/87
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6. EFFECTS
6.1. SYSTEMIC TOXICITY
6.1.1. Inhalation Exposure. Pertinent data regarding effects from
Inhalation exposure to 2-, 3- or 4-chloroanlllne could not be located In the
available literature as cited 1n Appendix A.
6.1.2. Oral Exposure.
6.1.2.1. SUBCHRONIC EXPOSURE Subchronlc range-finding studies were
conducted with F344 rats and B6C3F1 mice to establish the dietary concentra-
tions of 4-chloroan1l1ne used In an NCI (1979) cardnogenldty bloassay
(Section 6.2.2.). Groups of five animals of each sex were maintained on
diets that contained 0, 70, 145, 315, 680 or J465 ppm (rats) and 0, 255,
550, 1180, 2550, 5500, 8080, 11,830 or 17,380 ppm (mice) of 4-chloroan1l1ne
for 4 weeks, followed by an observation period of 2 weeks. Body weights and
food consumption were recorded twice weekly throughout the study, and all
*
survivors were necropsled upon termination of the study. It was not
Indicated 1f the necropsies Included hlstologlcal examinations. In rats,
survival was unaffected by chemical exposure and there were no treatment-
related effects on body weight gain. Enlarged spleens with plaque formation
occurred In all rats of both sexes at 680 and 1465 ppm but In none of the
rats at lower doses. All male and female mice treated with 8080 ppm and 4/5
males treated with 17,380 ppm died, but no cause of death was recorded;
survival was unaffected 1n the other groups. There were no clear treatment-
related effects on body weight gain In the mice. Enlarged spleens occurred
1n all 11,830 ppm male and all 17,380 female mice, but 1n none of the other
treated or control mice.
0028d -29- 06/05/87
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The abstract of a Russian study Indicated that oral administration of
3-chloroan1l1ne at 0.1-0.2 the L05Q for 30 days produced methemoglobln,
n1trosylhemoglob1n and sulfhemoglobin (Vasllenko et al., 1972). Additional
Information, Including the LD5Q, was not reported.
Pertinent data regarding the effects of subchronlc oral exposure to
2-chloroan1l1ne could not be located 1n the available literature as cited In
Appendix A.
6.1.2.2. CHRONIC EXPOSURE Several nonneoplastlc effects were
attributed to 4-chloroan1l1ne treatment 1n an NCI (1979) cardnogenldty
bloassay. Dietary concentrations of 250 or 500 ppm were administered to
F344 rats and 2500 and 5000 ppm to B6C3F1 mice for 78 weeks, followed by 24
*
weeks (rats) and 13 weeks (mice) of observation (Section 6.2.2.). As
detailed In Section 6.2.2., survival was reduced In the high-dose male rats,
and mean body weight gain was slightly depressed 1n the high-dose female
rats. There were no treatment-related effects on survival 1n the mice, but
mean body weights were reduced markedly 1n the high- and low-dose mice of
both sexes. Treatment-related nonneoplastlc lesions Included flbrosls of
the splenic capsule with subcapsular mesenchymal proliferation In male rats
(0/20, 45/49 and 38/49 In the control, low- and high-dose groups, respec-
tively) and female rats (0/20, 30/48, 43/50). Splenic neoplasms (Section
6.2.2.) appeared to arise 1n areas of capsular or parenchyma 1 flbrosls In
the rats. In mice, there was moderate to heavy Intracellular deposition of
Iron-positive pigment 1n most tissues, particularly the spleen, liver and
kidney, 1n both males (0/20, 35/50, 37/50) and females (0/18, 35/49, 33/42).
This pigment was Interpreted as hemoslderln, which probably resulted from
excessive compound-Induced hemolysls.
0028d -30- 05/15/87
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Pertinent data regarding the effects of chronic oral exposure to 2- and
3-chloroan1l1ne could not be located 1n the available literature as dted 1n
Appendix A.
6.1.3. Other Relevant Information. A single dose gavage LD5Q of 0.31
g/kg was determined for 4-chloroan1l1ne with Carworth-Wlstar male rats
(Smyth et a!., 1962).
Methemoglobln percentages were determined In cats hourly for 5 hours
following administration of single oral doses of 0.25 mmol/kg 2-chloro-
anlUne (31.9 mg/kg), 0.25 mmol/kg 3-chloroan1l1ne (31.9 mg/kg) or 0.0625
mmol/kg 4-chloroan1l1ne (8.0 mg/kg) (McLean et al., 1969). Groups of five
animals were assayed, and the overall means of the five posttreatment hourly
mean methemoglobln percentages were 53.1, 47.3 and 45.2 for 2-, 3- and
4-chloroanlllne, respectively, which Indicated that the 4-chloro Isomer 1s
the most potent methemoglobln Inducer. The Investigators did not state why
- * _
they administered such a low dose of 4-chloroan1l1ne relative to the other
Isomers. Cats (adult) were used 1n this study because they are particularly
sensitive to methemoglobln formation. Aniline 1s a recognized methemoglobln
Inducer (U.S. EPA, 1984), and many substituted anilines produced methemo-
globln 1n cats (McLean et al., 1969); aniline compounds 1n general are
regarded as potential methemoglobln Inducers (U.S. EPA, 1984). Abstracts of
Russian studies Indicate that sulfhemogloblnemla as well as hemogloblnemla
were effects of single or several dally 1ntraper1toneal or subcutaneous
Injections of 2-, 3- or 4-chloroan1l1ne 1n rats or mice (Vasllenko and
Zvezdal, 1972; Zvezdal, 1972; Nomura, 1975, 1980).
Single 1ntraper1toneal Injections of 1.0 mmol/kg (127.6 mg/kg) of
2-chloroan1l1ne or 1.5 mmol/kg (191.4 mg/kg) of 3- or 4-chloroan1l1ne were
nephrotoxlc to male F344 rats (Rankln et al., 1986). Effects Included
0028d -31- 06/05/87
-------
decreased urine volume, elevated BUN concentration and decreased basal and
lactate-stlmulated p-am1noh1ppurate accumulation by renal cortical slices.
6.2. CARCINOGENICITY
6.2.1. Inhalation. Pertinent data regarding the cardnogenlcHy of
Inhaled 2-, 3- or 4-chloroan1l1ne could not be located 1n the available
literature as cited 1n Appendix A.
6.2.2. Oral. The cardnogenlcHy of 4-chloroan1l1ne was evaluated 1n a
feeding study using Fischer 344 rats and B6C3F1 mice (NCI, 1979). Results
from this assay were also reported by Ward et al.(1980), Ward and Resnlk
(1981) and Goodman et al. (1984). Technical-grade compound was administered
to groups of fifty -6-week-old animals of each sex at diet concentrations of
250 or 500 ppm (rats) and 2500 or 5000 ppm (mice) for 78 weeks, followed by
observation periods of 24 weeks (rats) and 13 weeks (mice). Twenty
untreated animals of each sex and species served as controls. Body weights
were recorded weekly for the first 6 weeks, every 2 weeks for the next 12
weeks and at monthly Intervals thereafter for the remainder of the study.
Necropsies Including comprehensive hlstologlcal examinations were conducted
on all animals that were moribund, had palpable masses that jeopardized
their health, were killed at the end of the study or were found dead during
the study.
Slight mean body weight depression occurred 1n the high-dose female rats
after week 40 of treatment, but treatment-related effects on body weight did
not occur 1n the males (NCI, 1979). There was a significant positive
association (p«0.0294, Tarone test) between dosage and mortality 1n the male
rats (reflecting reduced high-dose survival after 60 weeks of treatment),
but not In the female rats. Adequate numbers of both male and female rats.
0028d -32- 06/05/87
-------
however, survived until the end of the study to be at risk for late-
developing tumors. Increased Incidences of splenic neoplasms, consisting
primarily of flbromas, occurred In the high-dose male rats. As detailed In
Table 6-1, there was a significant positive trend (p=0.001) between compound
administration and Incidences of pooled splenic tumors (Cochran-ArmHage
test) In the males, and the Incidence 1n the high-dose group was signifi-
cantly Increased (p=0.02) when compared with Incidences 1n the controls
(Fisher Exact test). There was a significant positive association between
dose and the combined Incidence of mesodermal cancers: flbromas, flbro-
sarcomas, hemanglosarcomas, osteosarcomas or sarcomas NOS of the spleen or
splenic capsule. Tumor Incidences were 0/20, 0/49 and 10/49 for the
control, low- and high-dose groups, respectively. However, the NCI report
notes that a metastaUc sarcoma not otherwise specified (NOS) was observed
1n the splenic capsule of one control male rat; however, no primary sarcoma
was detected. If 1t 1s assumed that the missing tumor was In the spleen of
this control rat thereby elevating the control Incidence for splenic tumors
to 1/20, making the control to high dose comparison no longer significant
(p=0.10) using the Fischer Exact Test. However, NCI also notes that the
historical Incidence of sarcomas NOS 1s 1/360 while the historic Incidence
of splenic flbromas, fIbrosarcomas, osteosarcomas and hemanglosarcomas Is
0/360. Therefore, NCI felt with these additional arguments that the data
suggests a compound-related Increase 1n the Incidence of splenic tumors for
the male rat.
In contrast to the rats, distinct mean body weight depression occurred
In dosed male and female mice (both dose groups) throughout the B6C3F1 mouse
study (NCI, 1979). There was no treatment-related mortality 1n either sex,
however, and adequate numbers of animals survived until the end of the study
0028d -33- 07/22/87
-------
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to be at risk for late-developing tumors. There was a significant positive
trend (p=0.01) between compound administration and Incidences of hemanglo-
sarcomas and combined Incidences of hemanglosarcomas and hemanglomas 1n the
female mice (see Table 6-1). The p value for the Fisher Exact test compar-
ing the combined Incidences of hemanglosarcomas and hemanglomas In the
female high-dose group with the Incidence 1n the control group was signifi-
cant at p-0.046. The combined Incidences of hemanglosarcomas and heman-
glomas In the male mice were dose-related (see Table 6-1), but neither the
Cochran-Armltage trend test nor the Fisher Exact Test were significant. The
NCI concluded that the above data are "Insufficient for correlating" the
occurrence of the hemanglomatous tumors to treatment. Consideration of the
f
"apparent" dose-related effect In both sexes and the historical Incidences
for combined hemanglosarcomas and hemanglomas In male and female B6C3F1 mice
(8/262 and 7/260, respectively; ~3X), however, suggests a positive assoda-
tlon between hemanglomatous tumors and p-chloroanlUne administration (NCI,
1979). There was also a positive trend (p=0.01) between compound adminis-
tration and combined Incidences of hepatocellular carcinoma and hepato-
cellular adenoma 1n the female mice (see Table 6-1), but the Fisher Exact
Tests were not significant. Moreover, the Incidences observed In the liver
were not unusual when compared with the historical control data.
NCI (1979) concluded that there was suggestive evidence In the male rat
and female and male mouse. The prollferatlve and neoplastlc lesions were
hlstologtcally Identical to those Induced 1n Fisher 344 rats by aniline
hydrochlorlde and o-tolu1d1ne hydrochlorlde.
The NTP Initiated a gavage bloassay of 4-chloroan1l1ne to further
clarify the results of the NCI (1979) study. This bloassay Involves
treatment of rats of both sexes at doses of 0, 2, 6 or 18 mg/kg/day and mice
0028d -36- 07/21/87
-------
of both sexes at doses of 0, 3, 10 and 30 mg/kg/day for 2 years (Canter,
1985). Preliminary findings Indicate that treatment with 4-chloroan1l1ne
Induced an Increased Incidence of splenic sarcomas In the male rats. The
study 1s currently 1s 1n the Pathology Working Group phase (MTP, 1987).
Pertinent data regarding the oral carclnogenldty of 2- or 3-chloroan1-
Une could not be located 1n the available literature as cited In Appendix A.
6.2.3. Other Relevant Information. 4-Chloroan1l1ne was tested 1n a
Strain A mouse pulmonary tumor assay with negative results (Maronpot et al.,
1986). In this test, groups of ten 6- to 8-week-old Strain A/St mice were
given 1ntraper1toneal Injections of 25, 57.5 or 60 mg/kg 1n trlcaprylln
vehicle, 3 times/week for 8 weeks; the two highest doses represented maximum
tolerated doses. Macroscopic evaluation of the lungs after a 16-week post-
treatment Incubation period showed that tumors occurred 1n only one of the
low-dose males. Survival was affected only In the male 57.5 mg/kg dose
group (4/10 were alive at study termination).
6.3. MUTAGENICITY
MutagenlcUy and other short-term assays have been conducted with 2-,
3- and 4-chloroan1l1ne. The results of these assays are detailed In Tables
6-2, 6-3 and 6-4 and are summarized briefly below.
2-Chloroan1l1ne produced reverse mutations In a study with Asperglllus
nldulans when tested without metabolic activation (Prasad, 1970) and In
studies of DNA repair In DNA repair deficient and proficient strains of
Escher1ch1a coll (pol A", pol A*) (Rosenkranz and Polrler, 1979; Po1r1er
and de Serres, 1979; Lelfer et al., 1981). Negative results were obtained
In reverse mutation assays with various strains of Salmonella typhlmurlum
when tested by different methods, 1n a mltotlc recombination assay with
Saccharomyces cerevlslae. In an unscheduled DNA synthesis assay with rat
0028d -37- 07/21/87
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hepatocytes and 1n a sperm head abnormality test with mice treated by
1ntraper1tonea1 Injection (see Table 6-2). Also, 2-chloroan1l1ne did not
produce neoplastlc transformation In cultured hamster embryo cells (see
Table 6-2).
3-Chloroan1l1ne produced reverse mutation In A. nldulans when tested
without metabolic activation (Prasad, 1970), but was not mutagenlc for
various strains of £. typhlmurlum when tested with or without activation and
did not produce unscheduled ONA synthesis 1n rat hepatocytes \n_ vitro (see
Table 6-3).
4-Chloroan1l1ne has been tested 1n numerous short-term genotoxlclty and
cell transformation assays. Positive responses occurred 1n a single reverse
mutation assay with A, nldulans (Prasad, 1970), 1n a single reverse mutation
assay with cultured L5178Y mouse lymphoma cells (NTP, 1983) and 1n DMA
repair assays with E_. coll (Lelfer et al., 1981; Rosenkranz and Polrler,
1979; Polrler and De Serres, 1979) and cultured rat hepatocytes (Williams et
al., 1982). Mostly negative results were reported In reverse mutation
assays with S. typhlmurlum (see Table 6-4). 4-Chloroan1l1ne produced
neoplastlc transformation 1n Rauscher leukemia virus-Infected rat embryo
cells (Traul et al., 1981; NTP. 1983) but elicited conflicting results with
hamster embryo cells (Plenta and Kawalek, 1981; Plenta. 1980) and negative
results with mouse BALB/c 3T3 cells (NTP, 1983).
The weight of evidence Indicates that the results of the short-term
assays of 2-, 3- and 4-chloroan1l1ne should be regarded as
Inconclusive.Positive responses were reported In several test systems, but
1n many cases these data are limited by conflicting results or a lack of
corroborating studies. Evaluation of the mutagenlcUy/clastogenldty of
2-chloroanlllne and 3-chloroanlllne Is further limited by a lack of variety
of assay types.
0028d -44- 07/21/87
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6.4. TERATOGEMICITY
Specific Information regarding the teratogenldty of the monochloro-
anlllnes could not be located 1n the available literature as cited In
Appendix A. The U.S. EPA (1984) has suggested, however, that aniline
compounds may cause teratogenldty by possible anoxia resulting from
methemoglob1nem1a.
6.5. OTHER REPRODUCTIVE EFFECTS
Specific Information regarding the reproductive effects of the mono-
chloroanlUnes could not be located 1n the available literature as cited 1n
Appendix A. The U.S. EPA (1984) believes, however, that aniline compounds
may cause adverse reproductive effects on the basis of Information that
repeated subcutaneous Injection of aniline Interferes With steroldogenesls
1n the rat uterus and that repeated oral administration of 4-chloro-3-n1tro-
anlllne causes sperm degeneration and an Increase In testes weights In rats.
6.6. SUMMARY
Pertinent data regarding the effects of Inhalation exposure to the
chloroanHlnes could not be located In the available literature as cited In
Appendix A. Effects of oral administration of chloroanlUnes are limited to
4-chloroan1l1ne. In a subchronlc dietary study of 4-chloroan1l1ne, rats
treated at >680 ppm had enlarged spleens with plaque formation; rats treated
at <380 ppm did not (NCI, 1979). Enlarged spleens also occurred 1n mice fed
at dietary levels >11,830 ppm. At 8080 ppm, all the mice died of unknown
causes. No effects were observed at 5500 ppm.
Based on an acute oral study using cats, 4-chloroan1l1ne was -4 times
more potent as a methemoglob1nem1a Inducer than were 2- or 3-chloroan1l1ne
(McLean et al., 1969).
0028d -45- 07/21/87
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4-Chloroan1l1ne was administered 1n the diet to rats at concentrations
of 250 or 500 ppm and mice at concentrations of 2500 and 5000 ppm for 78
weeks, followed by observation periods of 24 and 13 weeks, respectively
(NCI, 1979). Effects In the rats Included reduced survival 1n the high-dose
males, reduced body weight gain In the high-dose females, flbrosls of the
splenic capsule with subcapsular mesenchymal proliferation In most of the
treated males and females, and Increased Incidences of splenic flbromas and
sarcomas In the high-dose males. Effects In the mice Included markedly
reduced body weight gain 1n the high- and low-dose males and females, 1ntra-
cellular deposition of hemoslderln 1n many tissues In most of the treated
males and females and hemanglomatous tumors In the treated males and
females. NCI (1979) concluded that these findings are Insufficient to
establish carclnogenldty of 4-chloroan1l1ne In either rats or mice. The
splenic tumors 1n the rats were considered strongly suggestive of carclno-
genldty, however, because of the rarity of these tumors In historical
controls. Furthermore, preliminary results of an NTP gavage study using
rats and mice Indicate an Induction of splenic tumors In male rats (Canter,
1985). 4-Chloroan1l1ne was Inactive 1n a Strain A mouse pulmonary tumor
assay (Haronpot et al., 1986).
Specific Information regarding the carclnogenldty of 2- or 3-chloroan1-
Une could not be located In the available literature as cited In Appendix A.
Evidence for the N-ox1dat1on of monochloroanllines (see Section 5.3.) and
methemoglobln Induction by monochloroanlUnes (McLean et al., 1969) provides
an Indication of potential carclnogenldty, however, as Induction of both
carclnogenldty and methemoblob1nem1a by aniline and substituted aniline
compounds Is attributed to the formation of N-oxIdlzed metabolites (U.S.
EPA, 1984).
0028d -46- 07/21/87
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GenotoxIcHy and cell transformation testing of 2-, 3- and 4-chloro-
anlllne produced positive responses In several assays. Conflicting results,
a lack of corroborating data and, In the case of 2- and 3-chloroan1l1ne, a
limited variety of assay types Indicate that evidence for genotoxlclty of
the chloroanHlnes should be regarded as Inconclusive.
Specific Information regarding the teratogenldty or other reproductive
effects of the monochloroanllines could not be located 1n the available
literature as cited In Appendix A; however, the potential for these effects
has been suggested by the U.S. EPA (1984) on the basis of possible anoxia
resulting from methemogloblnemla and reproductive effects produced by other
aniline compounds.
0028d -47- 07/21/87
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7. EXISTING GUIDELINES AND STANDARDS
7.1. HUMAN
Pertinent guidelines and standards, Including EPA ambient water and air
quality criteria, drinking water standards, FAO/WHO ADIs, EPA or FDA
tolerances for raw agricultural commodities or foods, and ACGIH, NIOSH or
OSHA occupational exposure limits could not be located In the available
literature as dted 1n Appendix A.
7.2. AQUATIC
Guidelines and standards for the protection of aquatic organisms from
the effects of chloroanlUnes could not be located In the available
literature as dted In Appendix A.
0028d -48- 06/05/87
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8. RISK ASSESSMENT
Chronic and subchronlc toxldty data were available only for 4-chloro-
anlllne; therefore, risk assessment values cannot be derived for 2- or
3-chloroan1l1ne. Furthermore, It 1s not appropriate to derive risk assess-
ments for 2- or 3-chloroan1l1ne by analogy to 4-chloroan1l1ne because their
dose-responses for toxic effects may be significantly different. For
example, an acute oral study 1n cats Indicated that 4-chloroanlllne was -4
times more potent as an Inducer of methemogloblnemla than were 2- and
3-chloroan1l1ne (McLean et a!., 1969).
8.1. CARCINOGENICITY
8.1.1. Inhalation. Pertinent data regarding' the carclnogenldty of
Inhaled 2-, 3- or 4-chloroan1l1ne could not be located In the available
literature as cited In Appendix A.
8.1.2. Oral. An NCI (1979) carclnogenldty bloassay was -conducted In
which groups of 50 F344 rats of each sex were maintained on diets containing
250 or 500 ppm 4-chloroan1l1ne for 78 weeks, followed by an observation
period of 24 weeks. Groups of 50 B6C3F1 mice of each sex were similarly
treated with 2500 or 5000 ppm 4-chloroanlllne for 78 weeks, followed by 13
weeks of observation. Twenty untreated animals of each sex and species
served as controls. As detailed 1n Section 6.2.2. and Table 6-1, there was
a positive association between treatment and the occurrence of Hbromas of
the spleen 1n rats (significant Cochran-ArmHage test); however, the
Incidence 1n the high-dose group was not significantly Increased when
compared with the control group (Fisher Exact test). There was a signifi-
cant positive trend (p«0.001) between dose and the combined Incidence of
flbromas, fIbrosarcomas, hemanglosarcomas, osteosarcomas or sarcomas NOS of
0028d -49- 07/17/87
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the spleen or splenic capsule. The Fisher Exact Test comparing the high-
dose group with the concurrent controls was also significant at p=0.02.
Tumor Incidences were 0/20, 0/49 and 10/49 for the control, low- and high-
dose groups, respectively. However, the NCI report notes that a metastatlc
sarcoma NOS was observed In the splenic capsule of one control male rat;
however, no primary sarcoma was detected. If 1t 1s assumed that the missing
tumor was 1n the spleen of this control rat elevating the control Incidence
for splenic tumors to 1/20, the control to high dose comparison would no
longer be significant using the Fisher Exact Test (p=0.10). However, NCI
also notes that the historical Incidence of sarcomas NOS Is 1/360 while the
historic Incidence of splenic flbromas, flbrosarcomas, osteosarcomas and
^
hemanglosarcomas 1s 0/360. Therefore, NCI felt that the observed data
suggests a compound-related Increase In the Incidence of splenic tumors.
There was a significant dose-related positive trend (p-0.01) for
hemangloma or hemanglosarcoma "at all sites In the female mice and the
Incidence In high-dose females was significantly greater (p-0.04) than 1n
the controls. The combined Incidences of hemanglosarcomas and hemanglomas
In the male mice were Increased 1n a dose-related manner, but neither the
trend test nor Fisher Exact comparisons were significant at the p=0.05 level.
The occurrence of flbromas and sarcomas In the spleens of the treated
male rats was considered strongly suggestive of carc1nogen1c1ty. Further-
more, preliminary results of a study by NTP In which rats and mice were
treated by gavage with 4-chloroan1l1ne Indicate that treatment Induced
splenic sarcomas 1n male rats (Canter, 1985). The hemanglomatous tumors In
the treated mice of both sexes may also have been associated with 4-chloro-
anlllne treatment In the NCI (1979) study, as these tumors showed a trend
and statistical significance In the female mouse.
0028d -50- 07/21/87
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Pertinent data regarding the oral carclnogenlcHy of 2- or 3-chloroan1-
Une could not be located In the available literature as cited In Appendix A.
8.1.3. Other Routes. 4-Chloroan1l1ne was Inactive In a Strain A mouse
pulmonary tumor assay (Maronpot et a!., 1986).
8.1.4. Weight of Evidence. The rat and mouse carclnogenldty bloassay
evidence combined with the Inconclusive mutagenldty data would suggest that
p-chloroanHlne 1s In the C to B2 range according to EPA's cancer risk
assessment guidelines. The positive male rat data with the positive female
mouse data gives two species, albeit almost marginally so In the mouse and
with some uncertainty In the rat.
The suggestion of a positive trend In the male mouse using historical
controls Is not Inconsistent with the other observations. The pathologic
observations are consistent with similar observations made by NCI with
aniline hydrochlorlde and o-toluldlne hydrochlorlde. Given that a repeat
bloassay 1s underway the results of which are qualitatively reported to be
similar for the male rat, a provisional Group C weight of evidence Is
assigned to 4-chloroan1l1ne, provisional to the extent that a review should
be done when the new bloassay Is finished to finalize the weight of
evidence. There are also reasons to consider a B2 ranking; however, this
should be reserved for the time when the results from the new NTP bloassay
can be utilized. The lack of carclnogenldty data for 2- and 3-chloroanl-
llne Indicates that these compounds should be categorized In EPA Group D.
8.1.5. Quantitative Risk Estimates.
8.1.5.1. INHALATION Data regarding the carclnogenldty of
4-chloroan1l1ne by the Inhalation route were not available. Precedent
exists for extrapolating from the Inhalation route to the oral route,
particularly when differential absorption data are available (U.S. EPA,
0028d -51- 07/22/87
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1980). However, extrapolation from the oral route to the Inhalation route
Includes the additional uncertainty of possible direct effects of the
Inhaled compound on the pulmonary system. In this Instance, since rela-
tively little Is known concerning the pharmacoklnetlcs and pharmacodynamlcs
of 4-chloroan1lk1ne, a formal extrapolation 1s not pursued.
8.1.5.2. ORAL The strong suggestion that 4-chloroan1l1ne was
carcinogenic In male rats and female mice provides a basis to evaluate a
q * for oral exposure. The data used to calculate the q * are presented
1n Appendix B. The transformed doses were calculated by multiplying the
dietary concentrations by 0.05 (assuming that a rat consumes a dally amount
of food equal to 5% of Its body weight) and by the ratio of the length of
*
exposure to the length of the experiment (78 weeks/102 weeks). The
unadjusted q * was calculated using the computerized multistage model
developed by Howe and Crump (1982). The human q,* of 3.5xlO~2
(mg/kg/day)"1 was calculated by multiplying the unadjusted q,* by the
cube root of the ratio of the reference human body weight (70 kg) to the
mean weight of the male rats {0.38 kg, estimated from the growth curves In
the study) and by the cube of the ratio of the Hfespan of the rat (104
weeks) to the length of the experiment (102 weeks).
Assuming that a 70 kg person drinks 2 I water/day, the concentrations
of 4-chloroan1l1ne 1n drinking water associated with an Increased lifetime
risk of cancer at risk levels of 10'5, 10~* and 10~7 are lxlO~a,
lxlO~a and 1x10"* mg/l, respectively.
For comparative purposes, a q,* value was calcualted using the
Incidence of hemanglomas and hemanglosarcomas 1n female mice. The data are
shown 1n Appendix B. A somewhat lower potency value of 6.5x10""
0028d .. -52- 07/21/87
-------
(mg/kg/day)"1 was obtained. Therefore, the higher value based upon the
rat data 1s chosen to approximate the potency of 4-chloroan1l1ne for the
purposes of this risk assessment.
The q,* should be regarded as provisional until the results of the NTP
study, which may provide more definitive evidence, become available.
8.2. SYSTEMIC TOXICITY
8.2.1. Inhalation Exposure. Pertinent data regarding the toxldty of
Inhaled 2-, 3- or 4-chloroan1l1ne could not be located In the available
literature as cited 1n Appendix A. Calculation of chronic or subchronlc
Inhalation RfOs are precluded by the lack of these data.
8.2.2. Oral Exposure.
8.2.2.1. LESS THAN LIFETIME EXPOSURES (SUBCHRONIC) Subchronlc
range-finding toxldty studies of 4-chloroanlllne were conducted with F344
rats and B6C3F1 mice as part of the NCI (1979) cardnogenldty bloassay.
Groups of five animals of each sex were maintained on diets that provided 0,
70, 145, 315, 680 or 1465 ppm (rats) and 0, 255, 550, 1180, 2550, 5500,
8080, 11,830 or 17,380 ppm (mice) of compound for 4 weeks, followed by an
observation period of 2 weeks. Necropsies conducted on survivors at the end
of the study showed that enlarged spleens with plaque formation occurred In
all rats of both sexes at <680 ppm, but 1n none of the rats at lower doses.
There were no treatment-related effects on rat body weight or survival. As
detailed 1n Section 6.1.2.1., mortality and enlarged spleens occurred In
mice of both sexes at <8080 ppm. There were no clear treatment-related
effects on body weight 1n the mice. The cause of the mortality 1n the mice
was not specified, necropsies were only conducted on surviving rats and mice
and 1t was not Indicated 1f the necropsies Included hlstologlcal exami-
nations.
0028d -53- 07/21/87
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8.2.2.2. CHRONIC EXPOSURES 4-chloroan1Hne showed evidence of
carclnogenldty 1n rats In the NCI (1979) study and a q * was derived. An
RfD could be based on nonneoplastlc effects 1n the NCI (1979) study. In
this study, groups of 50 F344 rats of each sex were treated In the diet at
concentrations of 250 or 500 ppm for 78 weeks, followed by 24 weeks of
observation. Groups of 50 B6C3F1 mice were similarly treated at concentra-
tions of 2500 or 5000 ppm for 78 weeks, followed by 13 weeks of observation.
Control groups consisted of 20 untreated animals of each sex and species.
As detailed 1n Section 6.2.2., survival was reduced In the high-dose male
rats, average body weight was decreased In the high-dose female rats and
average body weights were decreased 1n both sexes of mice at both dose
levels. Treatment-related nonneoplastlc lesions of the capsule of the
spleen (focal flbrosls with subcapsular mesenchymal proliferation) occurred
In most of the treated rats (see Section 6.1.2.2.). Moderate to heavy
Intracellular deposits of pigment occurred In many tissues, particularly the
spleen, liver and kidney, 1n most of the treated mice. This pigment was
thought to be hemoslderln, resulting from excessive treatment-Induced
hemolysls. The dietary levels of 250 ppm 1n the rat study and 2500 ppm 1n
the mouse study therefore represent LOAELs.
If 1t Is assumed that dally food consumption In the rats and mice Is 5
and 13% of body weight, respectively, then the LOAELs are equivalent to 12.5
and 325 mg/kg/day, respectively. However, 1n view of the evidence for
carclnogenldty, albeit limited, an RfD 1s not estimated. The final results
of the NTP study may provide more definitive evidence, as preliminary
results suggest that rats treated by gavage developed spleen sarcomas.
0028d -54- 07/21/87
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9. REPORTABLE QUANTITIES
9.1. BASED ON SYSTEMIC TOXICITY
An NCI (1979) carc1nogen1c1ty bloassay was conducted In which groups of
20, 50 and 50 F344 rats of each sex were exposed to 0, 250 and 500 ppm
4-ch1oroan111ne In the diet, respectively, for 78 weeks, followed by 24
weeks of observation. Groups of 20, 50 and 50 B6C3F1 mice of each sex were
exposed similarly to 0, 2500 and 5000 ppm, respectively, for 78 weeks,
followed by 13 weeks of observation. As detailed 1n Section 6.2.2. and
summarized In Table 9-1, nonneoplastlc effects 1n the rats Included reduced
survival 1n the high-dose males, decreased body weight gain In the high-dose
females and nonneoplastlc proHferatlve lesions of the spleen In most
animals of both sexes at both doses. Effects In the mice Included decreased
body weight gain and Intracellular deposits of pigments 1n many tissues In
both sexes at both doses. This pigment was particularly evident In the
spleen, liver and kidney and was thought to be hemoslderln resulting from
excessive treatment-related hemolysls.
The most severe effects, which are considered for RQ determination, are
decreased survival and splenic lesions. The lowest equivalent human doses
at which these effects occurred are 4.4 and 1.9 mg/kg/day, respectively (see
Table 9-1). Multiplication of these doses by 70 kg yields MEOs of 308
mg/day for decreased survival and 133 mg/day for splenic lesions, and the
RV.s are calculated to be 1.8 and 2.3, respectively. Pronounced life
shortening warrants an RV of 10. Selection of an RV for the splenic
c C
lesions Is complicated because organ function was not evaluated. An RV
of 6, typically reflecting pathologic alterations without a detectable
decrement In organ function, 1s assigned. Multiplication of the RV.s by
0028d -55- 06/05/87
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the RVgs yields CSs of 18 for life shortening and 14 for the splenic
lesions (Table 9-2). Both of these CSs correspond to an RQ of 1000, but the
higher CS 1s the preferred value (Table 9-3).
Calculation of RQs for 2- or 3-chloroan1l1ne are precluded by a lack of
relevant toxldty data (Table 9-4). Furthermore, It 1s not appropriate to
derive RQs for 2- and 3-chloroan1l1ne by analogy to 4-chloroan1l1ne because
dose-responses for their toxic effects may be significantly different. For
example, an acute oral study using cats Indicated that 4-chloroan1l1ne was
-4 times more potent as an Inducer of methemogloblnemla than were 2- or
3-chloroan1l1ne.
9.2. BASED ON CARCINOGENICITY
There was evidence that 4-chloroan1l1ne was carcinogenic In male rats
and female mice 1n the NCI (1979) study, placing 4-chloroan1l1ne 1n EPA
Group B2 to C range (see Section 8.1.). While some might consider a B2
category appropriate, the expectation of results from a repeat NTP bloassay
due In early 1988 should be used to formulate a final weight of evidence
classification. At present the compound 1s considered to be a provisional
Group C. The response data provides a basis to derive an RQ based on
carclnogenlclty. An F factor was calculated using the data 1n Table 9-5.
The method for calculating the transformed doses was given In Section
8.1.5.2. The unadjusted 1/ED10 was calculated using the multistage model.
The F factor of 4.1X10"1 (mg/kg/day)"1 was calculated by multiplying the
unadjusted 1/ED,Q by the cube root of the ratio of the reference human
body weight (70 kg) to the body weight of the rat (0.38 kg) and by the cube
of the ratio of the Hfespan of the rat (104 weeks) to the duration of the
study (102 weeks). For comparative purposes, an adjusted 1/ED-m
0028d -57- 07/21/87
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TABLE 9-2
Oral Composite Scores for 4-Chloroan1l1ne Using the Rat*
Animal Dose
(mg/kg/day)
Chronic
Human MED
(mg/day)
RVd
Effect
RVe CS RQ
25
12.5
308 1.8 reduced survival 10 18 1000
133 2.3 nonneoplastlc lesions 6 14 1000
In the spleen
*Source: NCI, 1979
0028d
-58-
07/17/87
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TABLE 9-3
4-Chloroan1l1ne
Minimum Effective Dose (MED) and Reportable Quantity (RQ)
Route: oral
Dose*: 308 mg/day
Effect: reduced survival
Reference: NCI, 1979
RVd: 1.8
RVe: 10
Composite Score: 18
RQ: 1000
*Equ1valent human dose
0028d -59- 07/17/87
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TABLE 9-4
2- and 3-Ch1oroan1l1ne
Minimum Effective Dose (MED) and Reportable Quantity (RQ)
Route:
Dose:
Effect:
Reference:
RVd:
RVe:
Composite Score:
RQ: Data are not sufficient for deriving an RQ.
0028d -60- 07/17/87
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TABLE 9-5
Derivation of Potency Factor (F) for 4-ChloroanHlne
Reference:
Exposure route:
Species:
Strain:
Sex:
Vehicle or physical state:
Body weight:
Duration of treatment:
Duration of study:
Llfespan of animal:
Target organ:
Tumor type:
Experimental doses/exposures (ppm):
Transformed doses (mg/kg/day):
Tumor Incidence:
Unadjusted 1/ED10:
Adjusted 1/ED-io (F Factor):
NCI, 1979
oral
rat
F344
male
diet
0.38 kg (measured)
78 weeks
102 weeks
104 weeks
spleen
f1broma/fIbrosarcoma
0 250
0 9.6
0/20 0/49
6.802xlO~2 (mg/kg/day)'1
4.102X10'1 (mg/kg/day)'1
500
19
10/49
0028d
-61-
07/17/87
-------
calculated from the Incidence of hemanglomas and hemanglosarcomas 1n female
mice. The resulting 1/ED,Q value was 6.2xlO~2. Because the F factor 1s
<1, 4-chloroanHlne Is placed 1n Potency Group 3. An EPA Group C chemical
In Potency Group 3 has a LOW hazard ranking under CERCLA; therefore, the RQ
based on carc1nogen1c1ty Is 100.
0028d -62- 07/21/87
-------
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4-chloroan1l1ne and 3,4-d1chloroan1l1ne In various soils. Z. Pflanzezer-
naehr. Boden kd. 141: 57-66.
0028d -79- 07/17/87
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Surovtseva, E.G., G.K. Vaslleva, A.I. Volnova and Yu.V. Shurukhln. 1977.
Acetylatlon of aniline and Us chloro-substHuted analogs by a Pseudomonas
aurantlaca culture. Dokl. Akad. Nauk. SSSR. 237: 220-223. (CA 88:18853g)
Thorn, N.S. and A.R. Agg. 1975. The breakdown of synthetic organic com-
pounds 1n biological processes. Proc. R. Soc. Lond. B. 189: 347-357.
Thompson, C.Z., L.E. H111, J.K. Epp and G.S. Probst. 1983. The Induction
of bacterial mutation and heatocyte unscheduled ONA synthesis by mono-
substituted anilines. Environ. Hutagen. 5(6): 803-811.
Topham, J.C. 1979. The detection of carcinogen-Induced sperm head abnor-
malities In mice. Mutat. Res. 69: 149-155.
Traul, K.A., K. Takayama, V. Kachevsky, R.J. Hlnk and J.S. Wolff. 1981.
Rapid In vitro assay for cardnogenldty of chemical substances 1n mammalian
cells utilizing an attachment-Independence endpolnt. 2. Assay validation.
J. Appl. Toxlcol. 1(3): 190-195.
Uehleke, H. 1967. Stimulation of several mlcrosomal drug oxidations by
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Uehleke, H., 0. Reiner and K.H. Hellmer. 1971. Perinatal development of
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0028d -80- 07/17/87
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U.S. EPA. 1980. Guidelines and Methodology Used In the Preparation of
Health Effect Assessment Chapters of the Consent Decree Water Quality
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U.S. EPA. 1983. Methodology and Guidelines for Reportable Quantity Deter-
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and Environmental Assessment, Environmental Criteria and Assessment Office,
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U.S. EPA. 1984. Aniline and Chloro-, Bromo- and/or NHroanlUnes; Response
to the Interagency Testing Committee. Federal Register. 49: 108-126.
U.S. EPA. 1986a. Methodology for Evaluating Cardnogenldty In Support of
Reportable Quantity Adjustments Pursuant to CERCLA Section 102. Prepared by
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Office of Solid Waste and Emergency Response, Washington, DC.
U.S. EPA. 1986b. Guidelines for Carcinogen Risk Assessment. Federal
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U.S. EPA. 1987a. SANSS (Structure and Nomenclature Search System)
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U.S. EPA. 1987b. Graphical Exposure Modeling System (GEMS). Fate of
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0028d -81- 07/17/87
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USITC (U.S. International Trade Commission). 1984. Imports of Benzenold
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Van Blade!, R. and A. Moreale. 1977. Adsorption of herbicide derived
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Vasllenko, N.H., V.I. Zvenzdal and I.I. Kovalenko. 1972. Inactlvatlon of
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0028d -82- 07/17/87
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Uegman, R.C.C. and G.A.L. Oekorte. 1981. Aromatic amines 1n surface waters
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Zepp, R.G. and P.P. Schlotzhauer. 1983. Influence of algae on photolysis
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Zeyer, J. and P.C. Kearney. 1982. Mlcroblal degradation of para-chloroanl-
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215-223.
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Zlmmer, D., J. Mazurek, G. Petzold and E.K. Bhuyan. 1980. Bacterial
mutagenlcUy and mammalian cell DNA damage by several substituted anilines.
Mutat. Res. 77(4): 317-326.
Zoeteman, B.C.J., K. Harmsen, J.B.H.J. Llnders, C.F.H. Horra and U. Slooff.
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Netherlands. Chemosphere. 9: 231-249.
Zvezdal, V.I. 1972. Comparative diagnostic value of various pathological
derivatives of hemoglobin In conditions of acute and subacute poisoning by
aniline, nitrobenzene and their chloride derivatives. Farmakol. Tokslkol.
7: 159-162.
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APPENDIX A
LITERATURE SEARCHED
This HEED Is based on data Identified by computerized literature
searches of the following:
TSCATS
CASR online (U.S. EPA Chemical Activities Status Report)
TOXLINE
TOXBACK 76
TOXBACK 65
RTECS
OHM TADS
STORET
SRC Environmental Fate Data Bases
SANSS
AQUIRE
TSCAPP
NTIS
Federal Register
These searches were conducted In January, 1987. In addition, hand searches
were made of Chemical Abstracts (Collective Indices 5-9), and the following
secondary sources should be reviewed:
ACGIH (American Conference of Governmental Industrial Hyg1en1sts).
1986. Documentation of the Threshold Limit Values and Biological
Exposure Indices, 5th ed. Cincinnati, OH.
ACGIH (American Conference of Governmental Industrial Hyglenlsts).
1986-1987. TLVs: Threshold L1mH Values for Chemical Substances In
the Work Environment adopted by ACGIH with Intended Changes for
1986-1987. Cincinnati, OH. Ill p.
Clayton, G.D. and F.E. Clayton, Ed. 1981. Patty's Industrial
Hygiene and Toxicology, 3rd rev. ed.. Vol. 2A. John Wiley and
Sons, NY. 2878 p.
Clayton, G.D. and F.E. Clayton, Ed. 1981. Patty's Industrial
Hygiene and Toxicology, 3rd rev. ed., Vol. 2B. John Wiley and
Sons, NY. p. 2879-3816.
Clayton, G.D. and F.E. Clayton, Ed. 1982. Patty's Industrial
Hygiene and Toxicology, 3rd rev. ed., Vol. 2C. John Wiley and
Sons, NY. p. 3817-5112.
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Grayson, M. and 0. Eckroth, Ed. 1978-1984. K1rk-0thmer Encyclo-
pedia of Chemical Technology, 3rd ed. John WHey 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. WHO, IARC, Lyons, France.
Jaber, H.M., W.R. Mabey, A.T. L1eu, T.M. Chou and H.L. Johnson.
1984. Data acquisition for environmental transport and fate
screening for compounds of Interest to the Office of Solid Waste.
SRI International, Menlo Park, CA. EPA 600/6-84-010. NTIS
PB84-243906.
NTP (National Toxicology Program). 1986. 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). 1986. Directory of Chemical
Producers.* Menlo Park, CA.
U.S. EPA. 1986. Report on Status Report In the Special Review
Program, Registration Standards Program and the Data Call 1n
Programs. Registration Standards and the Data Call 1n Programs.
Office of Pesticide Programs, Washington, DC.
U.S. EPA. 1985. CSB Existing Chemical Assessment Tracking System.
Name and CAS Number Ordered Indexes. Office of Toxic Substances,
Washington. DC.
USITC (U.S. International Trade Commission). 1985. Synthetic
Organic Chemicals. U.S. Production and Sales, 1984, USITC Publ.
1422, 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.
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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. Flnley. 1980. Handbook of Acute Toxldty
of Chemicals to F1sh and Aquatic Invertebrates. Summaries of
Toxldty Tests Conducted at Columbia National Fisheries Research
Laboratory. 1965-1978. U.S. Dept. Interior, Fish and Wildlife
Serv. Res. Publ. 137, Washington, DC.
McKee, J.E. and H.W. Wolf. 1963. Water Quality Criteria, 2nd ed.
Prepared for the Resources Agency of California, State Water
Quality Control Board. Publ. No. 3-A.
Plmental, D. 1971. Ecological Effects of Pesticides on Non-Target
Species. Prepared for the U.S. EPA, Washington, DC. PB-269605.
Schneider, B.A. 1979. Toxicology Handbook. Mammalian and Aquatic
Data. Book 1: Toxicology Data. Office of Pesticide Programs, U.S.
EPA, Washington, DC. EPA 540/9-79-003. NTIS PB 80-196876.
0028d -87- 07/17/87
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APPENDIX B
Cancer Data Sheet for Derivation of q-j*
Compound: 4-Chloroan1l1ne
Reference: NCI, 1979
Specles/straln/sex: rat, F344, M
Route/vehicle: oral, diet
Length of exposure (1e) = 78 (weeks)
Length of experiment (Le) * 102 (weeks)
Llfespan of animal (L) * 104 (weeks)
Body weight » 0.38 kg (measured)
Tumor site and type: spleen and spleen capsule, fibroma, flbrosarcoma,
hemanglosarcoma, osteosarcoma or sarcoma NOS
Experimental Doses
or Exposure (ppm)
0
250
500
Transformed Dose
(mg/kg/day)
0
9.6
19.1
Incidence
No. Responding/No.
0/20
0/49
10/49
Tested
Unadjusted q-|* - 5.78215x10'" (mg/kg/day)'1
Human q-j* « 3.48733x10"* (mg/kg/day)'1
U.S. Environmental Protection Agency
Region V, Library
230 South Dearborn Street
0028d -aa^hicago, Illinois 60604 07/17/87
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APPENDIX B (cont.)
Cancer Data Sheet for Derivation of q-j*
Compound: 4-Chloroanlllne
Reference: NCI, 1979
Specles/straln/sex: mouse, B6C3F1, F
Route/vehicle: oral, diet
Length of exposure (le) = 78 (weeks)
Length of experiment (Le) - 91 (weeks)
Llfespan of animal (L) = 91 (weeks)
Body weight = 0.028 kg (measured)
Tumor site and type: hemanglosarcoma or hemangloma, all sites
Experimental Doses Transformed Dose
(ppm) (mg/kg/day)
Equivalent
Human Dose Incidence
(mg/kg/day) No. Responding/No. Tested
0
2500
5000
0
279
557
0
20.6
41.1
0/18
3/49
8/42
Human q-|* = 6.5xlO"» (mg/kg/day)"1
0028d
-89-
07/21/87
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