UHAM
United States : FCAO-CTN-GQflq
Env.ronmental Protection « ,i««
Agency May, 1990
&EPA Research and
Development
HEALTH AND ENVIRONMENTAL EFFECTS DOCUMENT
FOR 2,4,6-TRINITROTOLUENE
Prepared for
OFFICE OF SOLID WASTE AND
EMERGENCY RESPONSE
Prepared by
Environmental Criteria and Assessment Office
Office of Health and Environmental Assessment
U.S. Environmental Protection Agency
Cincinnati, OH 45268
DRAFT: DO NOT CITE OR QUOTE
NOTICE
Dcument 1s a preliminary draft. It has not been formally released
. Environmental Protection Agency and should not at this stage be
to represent Agency policy. It 1s being circulated for comments
mica! accuracy and policy Implications.
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DISCLAIMER
This report Is an external draft for review purposes only and does not
constitute Agency policy. Mention of trade names or commercial products
does not constitute endorsement or recommendation for use.
11
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PREFACE
Health and Environmental Effects Documents (HEEDs) are prepared for the
Office of Solid Waste 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 for Agency Program Office files are evaluated as they
pertain to potential human health, aquatic life and environmental effects of
hazardous waste constituents. The literature searched for 1n this document
and the dates searched are Included In "Appendix: Literature Searched."
Literature search material 1s current up to 8 months previous to the final
draft date listed on the front cover. Final draft document dates (front
cover) reflect the date the document 1s sent to the Program Officer (OSWER).
Several quantitative estimates are presented provided sufficient data
are available. For systemic toxicants, these Include Reference doses (R't'Ds)
for chronic and subchronlc exposures for both the Inhalation and oral
exposures. The subchronlc or partial lifetime RfD Is an estimate of an
exposure level that would not be expected to cause adverse effects when
exposure occurs during a limited time Interval I.e., for an Interval that
does not constitute a significant portion of the llfespan. This type of
exposure estimate has not been extensively used, or rigorously defined as
previous risk assessment efforts have focused primarily on lifetime exposure
scenarios. Animal data used for subchronlc estimates generally reflect
exposure durations of 30-90 days. The general methodology for estimating
subchronlc RfDs Is the same as traditionally employed for chronic estimates,
except that subchronlc data are utilized when available.
In the case of suspected carcinogens, a carcinogenic potency factor, or
q-|* (U.S. EPA, 1980) 1s provided. These potency estimates are derived for
both oral and Inhalation exposures where possible. In addition, unit risk
estimates for air and drinking water are presented based on Inhalation and
oral data, respectively. An RfO may also be derived for the noncarclnognelc
health effects of compounds that are also carcinogenic.
Reportable quantities (RQs) based on both chronic toxldty and carclno-
genldty are derived. The RQ 1s used to determine the quantity of a hazard-
ous substance for which notification Is required 1n the event of a release
as specified under the Comprehensive Environmental Response, Compensation
and Liability Act (CERCLA). These two RQs (chronic toxldty and cardno-
genldty) represent two of six scores developed (the remaining four reflect
IgnHabUHy, reactivity, aquatic toxldty, and acute mammalian toxldty).
Chemical-specific RQs reflect the lowest of these six primary criteria. The
methodology for chronic toxldty and cancer based RQs are defined 1n U.S.
EPA, 1984 and 1986a, respectively.
111
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EXECUTIVE SUMMARY
Trinitrotoluene, commonly referred to as TNT, Is a yellow crystalline
solid at room temperature. It 1s soluble In alcohol, ether, acetone,
benzene and carbon dlsulflde and slightly soluble In water (Sax and Lewis,
1987; Wlndholz et al., 1983). Between 3.1 and 31.1 million pounds of tri-
nitrotoluene was produced or Imported In the United States In 1977 at six
different facilities (TSCAPP, 1989). Earlier production data list the
monthly production of trinitrotoluene In the United States at -45 million
pounds during 1969-1971 (Ryo'n et al., 1984). Trinitrotoluene Is produced by
the nitration of toluene In a concentrated mixture of sulfurlc and nitric
adds (Ryon et al., 1984). It Is used chiefly as a high or bursting
explosive and as an Intermediate 1n dyestuffs and photographic chemicals
(Sax and Lewis, 1987).
The dominant fate process for trinitrotoluene In water Is expected to be
destruction by direct photolysis. The half-life for the sunlight photolysis
of trinitrotoluene In pure water Is 14 hours over the continental United
States during the summer months (Mabey et al., 1983). The sunlight photo-
lysis of trinitrotoluene In distilled water at room temperature occurred at
a similar rate with a half-life of 15 hours (Burllnson et al., 1973). The
rate was nearly pH-1ndependent, Increased with the presence of humlc
material, and decreased 1n the presence of oxygen and other triplet
quenchers (Mabey et al., 1983). The biological degradation of trinitro-
toluene In environmental waters 1s expected to occur under both aerobic and
anaerobic conditions, although the presence of additional nutrients may be
required (Carpenter et al., 1978; Osmon and Klausmeler, 1973; Chambers et
al., 1963; Tabak et al., 1964; Enzlnger, 1970; Spanggord et al., 1981, 1983;
1v
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Fewson, 1981; Hoffsommer et al., 1978). Products from both the aerobic and
anaerobic degradation of trinitrotoluene are believed to come from Initial
reduction of one or more of the nltro groups, which are then capable of
undergoing nonenzymatlc reactions, which may result In the formation of
products that are degradation-resistant. Complete degradation of trinitro-
toluene to CO. has. not been observed. Neither hydrolysis nor volatiliza-
tion to the atmosphere are expected to be significant In water. Adsorption
to sediment and suspended organic matter may occur, although it Is not
expected to be a significant process (Ryon et al., 1984). B1oconcentrat1on
In fish and aquatic organisms 1s not expected to be a significant process.
If released to soil. m1croblal degradation 1s expected to occur (Kaplan and
Kaplan, 1982a; Chambers et al., 1963; Tabak et al.. 1964; Osmon and
Klausmeler, 1973). The potential for strong adsorption and, thus, low
mobility 1n soil has been demonstrated by Kayser and Burllnson (1988).
Volatilization from the soil surface to the atmosphere 1s not expected to be
significant. In the atmosphere, trlnltro- toluene 1s expected to exist In
both the vapor phase and the partlculate form (Jones, 1960; Pella, 1977;
E1senre1ch et al., 1981). No experimental data on the direct photolytlc
degradation of atmospheric trinitrotoluene were located In the available
literature; however, this compound reportedly undergoes light-Induced
decomposition In the solid state (Burllnson et al., 1973). Therefore,
direct photolysis of gaseous and partlculate trinitrotoluene may be a
significant process 1n the atmosphere. No experimental data on the physical
removal of trinitrotoluene were located; however, It may be deposited on the
earth's surface by rain and partlculate settling (Ryon et al., 1984).
Limited data on exposure to trinitrotoluene were located In the avail-
able literature cited In Appendix A. Dermal exposure for those working In
areas related to the production of armaments 1s reportedly more significant
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than exposure by Inhalation (Woollen et al., 1986). Trinitrotoluene was
detected In groundwater and surface water samples near sites of Us produc-
tion or manipulation (Jenkins et al., 1986; Nay et al., 1972; Spanggord et
al., 1982; Perelra et al., 1979; Spaldlng and Fulton, 1988). Thus, the
potential for 1ngest1on of this compound exists for a small group of the
population, although quantitative estimations of human exposure for the
group cannot be made because of the lack of drinking water and food monitor-
Ing data.
Static LC.. values for 2,4,6-tr1n1trotoluene varied from 5.2-27.0
mg/l among Invertebrates and 0.8-3.4 mg/l among fish (Liu et al., 1983;
Pederson, 1970). In flowthrough tests, LC,Q values among fish ranged from
2.0-3.7 mg/l (L1u et al., 1983; Smock et al., 1976). Daphnla maqna was
the most sensitive species In flowthrough tests, with a 96-hour LC,-n of
1.2 mg/l and an Incipient LC5Q of 0.19 mg/l at 192 hours (Liu et al.,
1983). In a 21-day static model ecosystem, the LOEC values for reduced
population size and Individual growth were 0.6 mg/l In Daphnla and 5.6
mg/l In the ollgochaete, LumbMculus varlegatus (Bailey, 1982). Decreased
algal density may have been responsible for the reduction In Daphnla popula-
tion. The population density of the green alga, Selenastrum caprlcornutum.
fell Immediately following exposure to 5.6 mg/l 1n this study. Algal
density also decreased In microcosms exposed to lower concentrations, but
this was a delayed effect that may have been due to growth of the Daphnla
population. Other studies on algae reported LOEC values of 4.1-5 mg/l 1n
Scenedesmus caprlcornutum. 1.6 mg/l 1n S. quadrlcauda. 0.32-25 mg/l In
Hlcrocystls aeruqlnosa. 4.1 mg/l In Anabaena flosaquae and 18 mg/l 1n
the diatom Navlcula pelllculosa (BMngmann and Kuhn, 1978; Fitzgerald et
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al., 1952; Liu et al., 1983; Smock et al., 1976). Photolysis of 2,4,6-tM-
nltrotoluene was reported In two of these studies and may have affected the
results 1n others. The LOEC for reduced growth was 1 mg/l In an aquatic
flowering plant, the duckweed, Lemna perpusllla. and 5 mg/l In a terres-
trial plant, the yellow nutsedge, Cyperus esculentus (Palazzo and Leggett,
1986; Schott and Worthley, 1974). Concentrations <100 mg/l had no effect
on cell multiplication In the bacterium, Pseudomonas putlda. Several
studies Included assays designed to determine the Influence of water quality
variables upon the toxldty of 2,4,6-tr1n1trotoluene In various species.
None reported more than slight changes attributable to these variables (Liu
et al., 1983; Pederson, 1970; Schott and Worthley, 1974). Exploratory 4-day
static bloconcentratlon studies reported BCFs ranging from 202-453 In
several representative Invertebrates, fish and algae.
In mice, rats, dogs and rabbits, trinitrotoluene administered orally,
dermally or Intratracheally (only to rats) Is readily absorbed, distributed,
metabolized and excreted In the urine and, to a lesser extent, In feces
(El-hawar1 et al., 1981). Generally, the rate of absorption by the three
routes tested was Intratracheal > oral > dermal. The extent of absorption
In the four species tested was not significantly different when trinitro-
toluene was administered by the oral route. After dermal exposure, however,
the highest absorption occurred In rabbits, followed by mice, rats and dogs.
Radioactivity was mainly distributed to the liver and kidney of the animals
after oral dosing, but fat contained appreciable amounts of radioactivity
following dermal treatment. Trinitrotoluene was extensively metabolized In
all species examined regardless of the route of administration. Identifica-
tion of products 1n the urine and bile showed that most metabolites were
nltroreductlon derivatives. Oxidation of the methyl group had also
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occurred. Unchanged trinitrotoluene could not be Identified In the urine of
rabbits. The metabolic profiles of urine from rats, mice and dogs and
different routes of exposure differed only quantitatively and no significant
sex differences were observed. Urine of rabbits was unique because 1t
contained larger amounts of monoamlnes and hydroxylamlnes. Urinary and
biliary excretion appear to play nearly equivalent roles In the elimination
of trinitrotoluene.
Reported oral LD5Qs for trinitrotoluene administered by gavage were
660 mg/kg In male and female mice and 1320 and 795 mg/kg In male and female
rats, respectively. LD5Q data for other species were not available. Data
were not available regarding the toxldty of trinitrotoluene to animals by
Inhalation exposure.
Data regarding the toxldty of trinitrotoluene In humans Indicated that
workers exposed to air levels between 0.01 and 4.0 mg/m3 may develop skin
Irritation, liver damage and anemia (Hathaway, 1977; Morton et al., 1976).
Although there are numerous reports of occupational exposure to trinitro-
toluene, the duration and levels of exposure were usually not sufficiently
defined to permit use of these studies In risk assessment.
Trinitrotoluene yielded evidence of carclnogenlclty In a 24-month
dietary exposure study using rats (Furedl et al., 1984a). In that study,
female rats had a significantly Increased Incidence of urinary bladder
paplllomas and carcinomas. Trinitrotoluene was not carcinogenic when tested
In mice {Furedl et al., 1984b). Trinitrotoluene was mutagenlc In reverse
mutation assays In Salmonella typhlmurlum In the absence of activating
systems (Kaplan and Kaplan, 1982b; Hhong and Edwards, 1984; Won et al.,
1976). The presence of activating systems reduced the mutagenlc potency.
Trinitrotoluene did not Induce DNA damage In mammalian test systems _Ui vivo
(Ashby et al., 1985).
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Subchronk studies using animals suggest that dogs are the most
sensitive species since a dose of 0.5 mg/kg/day for 26 weeks Induced signs
of anemia and liver alterations (Levlne et al., 1983). Increasing doses
Increased the severity of the effects. Death occurred with a dose of 32
mg/kg/day before week 17. In contrast, death from anemia occurred In rats
with doses of 300 mg/kg/day (-10 times higher than In dogs) administered for
13 weeks (Levlne et al., 1984a). Mice appeared to be less sensitive. A
dose of 190 mg/kg/day for 13 weeks produced liver effects, but a dose of 36
mg/kg/day was without adverse effects; It Is possible that doses between 36
and 190 mg/kg/day could have been toxic. Chronic studies have been
performed only on rats and mice. In rats, a dose of 2.0 mg/kg/day In the
diet for 24 months caused kidney hypertrophy, spleen congestion and bone
marrow Mbrosls In females. A dose of 10 mg/kg/day Induced signs of anemia,
changes 1n organ weights and urinary bladder lesions In females. In
contrast, a dose of 10 mg/kg/day administered In the diet for 24 months to
mice was without adverse effects. The most sensitive endpolnts for
assessing toxlcologlcal effects of trinitrotoluene seem to be the liver and
elements In the blood, such as the RBCs.
Data regarding the developmental or reproductive toxlclty of
trInltroluene were not available In the literature dted In Appendix A.
Based on the weight of evidence, trinitrotoluene has been assigned to
U.S. EPA Group C: possible human carcinogen. A q^ value of 3xlO"2
(mg/kg/day)"1 was previously derived (U.S. EPA, 1988b, 1989) from the
dose-response data for Increased Incidences of urinary bladder paplllomas
and carcinomas In female rats treated with trinitrotoluene In the diet
(Furedl et al., 1984a). The concentration of trinitrotoluene In drinking
water associated with a risk level of 1E-5 Is 10 wg/l. A carclnogen-
1c1ty-based RQ of 100 was assigned.
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An RfD of 5E-4 mg/kg/day was derived for subchronlc and chronic oral
exposure to trinitrotoluene based on the LOAEL of 0.5 mg/kg/day for liver
effects In dogs In the 26-week oral study by Levlne et al. (1983). An RQ of
100 for chronic (noncancer) toxldty was derived based on liver effects 1n
dogs (Levlne et al., 1983).
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TABLE OF CONTENTS
Page
1. INTRODUCTION 1
1.1. STRUCTURE AND CAS NUMBER 1
1.2. PHYSICAL AND CHEMICAL PROPERTIES 1
1.3. PRODUCTION DATA 2
1.4. USE DATA 2
1.5. SUMMARY 2
2. ENVIRONMENTAL FATE AND TRANSPORT 4
2.1. AIR 4
2.1.1. Reaction with Hydroxyl Radicals 4
2.1.2. Reaction with Ozone 4
2.1.3. Photolysis 4
2.1.4. Physical Removal Processes 5
2.2. WATER 5
2.2.1. Hydrolysis 5
2.2.2. Oxidation 5
2.2.3. Photolysis 5
2.2.4. Mlcroblal Degradation 6
2.2.5. B1oconcentrat1on 8
2.2.6. Adsorption 9
2.2.7. Volatilization 9
2.3. SOIL 9
2.3.1. Mlcroblal Degradation 9
2.3.2. Adsorption 10
2.3.3. Volatilization 10
2.4. SUMMARY 10
3. EXPOSURE 13
3.1. HATER 13
3.2. FOOD 13
3.3. INHALATION 14
3.4. DERMAL 14
3.5. OTHER 14
3.6. SUMMARY 14
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TABLE OF CONTENTS (cont.)
4. ENVIRONMENTAL TOXICOLOGY 16
4.1. AQUATIC TOXICOLOGY 16
4.1.1. Acute Toxic Effects on Fauna 16
4.1.2. Chronic Effects on Fauna 18
4.1.3. Effects on Flora 19
4.1.4. Effects on Bacteria 21
4.2. TERRESTRIAL TOXICOLOGY 22
4.2.1. Effects on Fauna 22
4.2.2. Effects on Flora 22
4.3. FIELD STUDIES 22
4.4. AQUATIC RISK ASSESSMENT 22
4.5. SUMMARY 24
5. PHARMACOKINETCS 26
5.1. ABSORPTION , 27
5.2. DISTRIBUTION 30
5.3. METABOLISM 34
5.4. EXCRETION 36
5.5. SUMMARY 37
6. EFFECTS . . 39
6.1. SYSTEMIC TOXICITY 39
6.1.1. Inhalation Exposure 39
6.1.2. Oral Exposure 39
6.1.3. Other Relevant Information 48
6.2. CARCINOGENICITY 51
6.2.1. Inhalation 51
6.2.2. Oral 51
6.2.3. Other Relevant Information 54
6.3. MUTAGENICITY 55
6.4. DEVELOPMENTAL TOXICITY 55
6.5. OTHER REPRODUCTIVE EFFECTS . . . . ' 55
6.6. SUMMARY 55
7. EXISTING GUIDELINES AND STANDARDS 59
7.1. HUMAN 59
7.2. AQUATIC 59
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TABLE OF CONTENTS (cont.)
Page
8. RISK ASSESSMENT 60
8.1. CARCINOGENICITY 60
8.1.1. Inhalation 60
8.1.2. Oral 60
8.1.3. Other Routes 61
8.1.4. Weight of Evidence 61
8.1.5. Quantitative Risk Estimates 61
8.2. SYSTEMIC TOXICITY 62
8.2.1. Inhalation Exposure 62
8.2.2. Oral Exposure 62
9. REPORTABLE QUANTITIES 66
9.1. BASEO ON SYSTEMIC TOXICITY 66
9.2. BASEO ON CARCINOGENICITY 70
10. REFERENCES 73
APPENDIX A: LITERATURE SEARCHED 87
APPENDIX B: SUMMARY TABLE FOR 2,4,6-TRINITROTOLUENE 90
APPENDIX C: DOSE/DURATION RESPONSE GRAPH(S) FOR EXPOSURE TO
2,4,6-TRINITROTOLUENE 91
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LIST OF TABLES
No. Title Page
5-1 Excretion of Radioactivity 24 Hours After Administration
of 14C-Tr1n1trotoluene 28
5-2 Tissue Distribution of Radioactivity 24 Hours After
Administration of l4C-Tr1n1troto1uene Using Rats 31
5-3 Tissue Distribution of Radioactivity 24 Hours After
Administration of l4C-Tr1n1trotoluene Using Rabbits 32
5-4 Tissue Distribution of Radioactivity 24 Hours After Oral
Administration of l4C-Tr1n1trotoluene Using Dog 33
6-1 Incidence of Urinary Bladder Tumors 1n Female F344 Rats fed
Diets Containing Trinitrotoluene (>99% pure) for 24 Months. . 52
6-2 Incidence of Hyperplastlc Lesions In F344 Rats Fed Diets
Containing Trinitrotoluene (>99X pure) for 24 Months .... 53
6-3 Mutagenlclty Testing of 2,4,6-TrlnHrotoluene 56
9-1 ToxKHy Summary for 2,4,6-TrlnHrotoluene 67
9-2 Composite Scores for Oral Toxlclty for 2,4,6-Trlnltrotoluene. 69
9-3 2,4,6-Tr1n1trotoluene: Minimum Effective Dose (MED) and
Reportable Quantity (RQ) 71
9-4 Derivation of Potency Factor (F) for 2,4,6-TrInltrotoluene. . 72
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LIST OF ABBREVIATIONS
BCF
BUN
CAS
CS
OMSO
ONA
EC50
PEL
F344
HA
K
ow
LC50
L°50
LDH
LOAEL
LOEC
MED
MTD
NOAEL
NOEC
NOEL
PEL
ppm
ppt
RBC
RfO
RQ
RVd
RV
e
SGPT
TLC
TLV
TWA
B1oconcentrat1on factor
Blood urea nitrogen
Chemical Abstract Service
Composite score
Dimethyl sulfoxlde
Oeoxyr1bonucle1c add
Concentration effective to 50% of recipients
Frank effect level
Fischer 344
Health advisory
Octanol/water partition coefficient
Concentration lethal to 50% of recipients
Oose lethal to 50% of recipients
Lactate dehydrogenase
Lowest-observed-adverse-effect level
Lowest-observed-effect concentration
Minimum effective dose
Maximum tolerated dose
No-observed-adverse-effect level
No-observed-effect concentration
No-observed-effect level
Permissible exposure level
Parts per million
Parts per trillion
Red blood cell
Reference dose
Reportable quantity
Dose-rating value
Effect-rating value
Serum glutamlc pyruvlc transamlnase
Thin layer chromatography
Threshold limit value
Time-weighted average
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1. INTRODUCTION
1.1. STRUCTURE AND CAS NUMBER
Trinitrotoluene Is the common name for 2,4,6-tr1n1trotoluene. It Is
also known as a-tr1n1trotoluene, sym or s-tr1nUrotoluene, 2-methyl-l,
3,5-tr1n1trobenzene, trllH, tolH, trUol and trinitrotoluene (Chemllne,
1989; SANSS, 1989). The structure, CAS Registry number, empirical formula
and molecular weight are as follows:
0,N
CAS number: 118-96-7
Empirical formula: C-.H-N-0,
/ b J b
Molecular weight: 227.13
1.2. PHYSICAL AND CHEMICAL PROPERTIES
Trinitrotoluene Is a yellow crystalline solid at room temperature. It
Is soluble 1n alcohol, ether, acetone, benzene and carbon dlsulflde and
slightly soluble In water (Sax and Lewis, 1987; Hlndholz et al., 1983).
Selected chemical and physical properties are given below.
Melting point:
Boiling point:
Density at 20°C:
Water solubility:
Log Kow:
Vapor pressure:
Conversion factor:
(air at 25'C)
80.2-81.3'C
186.6°C at 7.5 mm Hg
1.654 g/mi
104-113 mg/i at 20°C
1.60
8.02x10"* mm Hg at 25°C
4.09x10"* mm Hg at 25°C
1 mg/tn3 = 0.108 ppm
1 ppm = 9.259 mg/m3
Pella, 1977
BoubUk et al., 1984
Wlndholz et al., 1983
Spanggord et al., 1983
Hansch and Leo, 1985
Pella, 1977
Jones, 1960
0240d
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09/12/89
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1.3. PRODUCTION DATA
During 1977, six U.S. plants manufactured or Imported between -3.1 and
31.1 million pounds of trinitrotoluene: IMC Plaza 1n Llbertyvllle, IL;
Unlroyal, Inc. In Jollet, IL; E. I. Oupont de Neumours and Co. In
Wilmington, DE; Chemical Systems Division In San Jose, CA; Volunteer Army
Ammunition Plant In Chattanooga. TN; and one plant with production Informa-
tion listed as confidential (TSCAPP, 1989). Earlier data 11st the produc-
tion of trinitrotoluene at -45 million pounds/month for 1969-1971 (Ryon et
al., 1984). Government policy dictates that all munitions plants be
government-owned, although most of the plants are operated by contractors
who are usually major chemical companies (Ryon et al.f 1984).
Trinitrotoluene Is manufactured by the stepwlse nitration of toluene In
either a batch or continuous operation. Toluene, nitric add and sulfurlc
acid are mixed together In the first step of a six-step process. The nitra-
tion products from each stage are passed on to the next, where progressively
higher temperatures and acid concentrations are used to maximize the yield
of the desired trlnHrated product. The crude trinitrotoluene Is purified
by treatment with sodium sulfHe, followed by recrystallzatlon (Ryon et al.,
1984).
1.4. USE DATA
Trinitrotoluene Is used as a high explosive In military armaments and as
an Intermediate 1n dyestuffs and photographic chemicals (Sax and Lewis,
1987).
1.5. SUMMARY
Trinitrotoluene, commonly referred to as TNT, 1s a yellow crystalline
solid at room temperature. It Is soluble In alcohol, ether, acetone,
benzene and carbon dlsulflde and slightly soluble 1n water (Sax and Lewis,
0240d -2- 11/09/89
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1987; Wlndholz et al.f 1983). Between 3.1 and 31.1 million pounds of
trinitrotoluene was produced or Imported In the United States 1n 1977 at six
different facilities (TSCAPP, -1989). Earlier production data 11st the
monthly production of trinitrotoluene In the United States at -45 million
pounds during 1969-1971 (Ryon et al., 1984). Trinitrotoluene Is produced by
the nitration of toluene 1n a concentrated mixture of sulfurlc and nitric
adds (Ryon et al., 1984). It Is used chiefly as a high, or bursting,
explosive and as an Intermediate 1n dyestuffs and photographic chemicals
(Sax and Lewis, 1987);
0240d -3- H/Ol/89
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2. ENVIRONMENTAL FATE AND TRANSPORT
2.1. AIR
Based on Us reported vapor pressures of 4.09x10"* and 8.02xlO~« mm
Hg at 25°C (Jones, 1960; Pella, 1977), trinitrotoluene Is expected to exist
partially 1n the vapor phase but predominantly In partlculate form In the
atmosphere (Elsenrelch et al., 1981).
2.1.1. Reaction with Hydroxyl Radicals. Using the estimation method of
Atkinson (1985), a rate constant of 1.46xlO~13 cmVmolecule-sec can be
obtained for the vapor phase reaction of photochemlcally produced HO- with
trinitrotoluene. If the average atmospheric H0« concentration Is 5x10*
molecules/cm3 (Atkinson, 1985), then the half-life for this reaction Is
-110 days. Since trinitrotoluene Is not expected to exist entirely In the
vapor phase In the atmosphere (Elsenrelch et al., 1981; Jones, 1960; Pella,
1977), the actual rate of destruction by this process Is expected to be
considerably slower. Therefore, removal of atmospheric trinitrotoluene by
the gas-phase destruction by photochemlcally produced H0» Is not expected
to be significant.
2.1.2. Reaction with Ozone. The gas-phase destruction of atmospheric
trinitrotoluene through the reaction with ozone Is not expected to be
significant (Atkinson, 1985).
2.1.3. Photolysis. Pertinent quantitative data regarding the photolysis
of trinitrotoluene In the atmosphere were not located In the available
literature cited 1n Appendix A. Trinitrotoluene, however, Is susceptible
to photochemical degradation (Section 2.2.3.), even In the solid state
(Burllnson et al., 1973). Therefore, direct photochemical degradation of
both vapor-phase and partlculate trinitrotoluene 1n the atmosphere may occur.
0240d -4- 09/12/89
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2.1.4. Physical Removal Processes. Pertinent data regarding the physical
removal of trinitrotoluene from the atmosphere were not located 1n the
available literature dted 1n Appendix A. Ryon et al. (1984) postulated
that It may be deposited on the earth's surface by partlculate settling or
rain deposition.
2.2. WATER
2.2.1. Hydrolysis. Limited experimental data were located In the litera-
ture concerning the hydrolysis of trinitrotoluene In the environment. In
the laboratory, trinitrotoluene underwent no observable hydrolysis In sea
water after 108 days at 25"C at a pH of -8.1 (Hoffsommer and Rosen, 1973).
Hydrolysis Is not expected to be significant for trinitrotoluene, since It
contains no readily hydrolyzable functional groups (Harris, 1982).
2.2.2. Oxidation. Pertinent data regarding the chemical oxidation of
trinitrotoluene In water were not located In the available literature dted
In Appendix A. It Is not, however, expected to be a significant fate
process.
2.2.3. Photolysis. The degradation of trinitrotoluene by direct photoly-
sis In water Is well documented qualitatively and quantitatively. This
process Is described as the source of the pink water problem associated with
the wastewater of armament production (Burllnson et al., 1973).
The half-life for the sunlight photolysis of trinitrotoluene In pure
water, as estimated by experiments 1n natural sunlight, was 14 hours at a
latitude of 20, 40 or 50° over the continental United States during the
summer months. In the winter months, the respective half-lives were
estimated to be 22, 45 and 85 hours. In these studies, pH had a minor
effect on the rate of photolysis. The products produced 1n this reaction
accelerated the rate of trinitrotoluene photolysis, and humlc acid 1n
0240d -5- 11/01/89
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natural waters Increased the rate of photolysis from one to two orders of
magnitude. Oxygen and other triplet quenchers decreased the rate (Mabey et
al., 1983).
The laboratory test-tube photolysis of 4.97 ppm trinitrotoluene In water
obtained from a waste lagoon on an Army ammunition plant resulted In final
concentrations of 3.29 and 2.34 ppm after 110 minutes (at 313 nm) and 240
minutes (at 366 nm), respectively. Photolysis of the same solution using
natural sunlight resulted In a concentration decrease to 2.49 ppm after 80
minutes (Spanggord et al., 1983). ,
The photolysis (>280 nm) of tMnltrotofuene In pure water at 60°C In a
continuous flow apparatus produced 1,3,5-tr1n1trobenzene, 2,6-d1n1tro-
anthranll, 2,4,6-trlnltrobenzaldehyde and 2,4,6-tr1n1trobenzon1tr1le,
however; 80% of the mass balance could not be accounted for. The sunlight
photolysis of trinitrotoluene In pure water at room temperature In a quartz
vessel was 75X complete In -30 hours. In addition to the products listed
above, four tetranltroazoxytoluenes were Isolated reportedly because of the
condensation of the Initial products (Burllnson et al., 1973).
2.2.4. Hlcroblal Degradation. No Initial carbon-14 labeled trinitro-
toluene could be detected after 3-5 days 1n aerated reactors using a sewage
sludge Inoculum. Very little radlolabeled C0_ (<0.5X) was found In these
experiments, Indicating that complete mineralization did not occur. The
authors concluded that one or more of the nltro groups In trinitrotoluene
was converted to the corresponding amlne, which then reacted with a
carboxylate group from the cellular matter of the medium, forming, poly-
amides. The formation of polyamldes, thought to resist degradation,
accounts for the lack of CO- production and 1s consistent with the
observed lack of destruction for the aromatic skeleton of trinitrotoluene
(Carpenter et al., 1978).
0240d -6- 11/01/89
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Trinitrotoluene at an Initial concentration of 100 mg/l completely
disappeared under aerobic conditions In 6 days using Inocula obtained from
sewage treatment plants, wastewater from an ordinance loading facility and
soil, pond or aquarium water, all of which had been exposed to trinitro-
toluene previously. Yeast extract was added to this experiment to serve as
a source of nutrients. When no yeast extract was added, mlcroblal degrada-
tion did not occur (Osmon and Klausmeler, 1973).
Microbes obtained from a sewage treatment plant can be acclimated and
grown successfully In the presence of high concentrations (29-100 ppm) of
trinitrotoluene, such that the normal respiration of other organic compounds
could occur. However, no evidence for the blodegradatlon of trinitrotoluene
was presented In this experiment. Pure cultures of bacteria, Zoogloea
ramlqera 115, grown on trinitrotoluene degraded 1t under aerobic conditions
(Enzlnger, 1970).
Trinitrotoluene undergoes reductive blotransformatlon under both aerobic
and anaerobic conditions. 4-Am1no-2,6-d1nltrotoluene and 2-am1no-4,6-d1-
nHrotoluene are metabolites. Trinitrotoluene at an Initial concentration
of 10 ppm did not degrade after 6 weeks under aerobic conditions In water
containing sediment obtained from the Searsvllle Pond, CA, or "with eutrophlc
water obtained from the Coyote Creek, CA. When the experiment was repeated
with the addition of 500 ppm yeast extract, the amount of trinitrotoluene
was reduced to a level below the limit of detection within 5 days. Mixed
cultures obtained from Waconda Bay, TN, and raised on 2,4-d1n1trotoluene did
not degrade trinitrotoluene under aerobic conditions. In the presence of
other nutrients such as yeast extract, degradation proceeded (Spanggord et
al., 1981).
0240d -7- 11/01/89
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Trinitrotoluene at an Initial concentration of 5 ppm degraded to 2 ppm
over a 90-day period under aerobic conditions In lagoon water obtained from
an Army ammunition plant. The addition of yeast extract Increased the rate
of blodegradatlon. In lagoon sediment and water samples, an Initial tri-
nitrotoluene concentration of 74 ppm was reduced to 59 ppm over the course
of 90 days under aerobic conditions. Under anaerobic conditions In the
lagoon water alone, trinitrotoluene underwent blotransformatlon, although at
a slower rate than seen for the aerobic degradation In this sample. Yeast
extract, again, Increased the rate of blodegradatlon (Spanggord et al.,
1983).
Trinitrotoluene at a concentration between 10 and 50 ppm underwent 97%
blodegradatlon when continuously fed Into an aerated oxidation ditch
containing activated sludge and a cornsteep nutrient. Metabolites from this
process, In addition to the amlno-nltrotoluenes mentioned above, were postu-
lated to be 2- or 2,4-hydroxylam1non1trotoluenes, although they were not
Isolated (Hoffsommer et al., 1978). These hydroxyamlnonlnltrotoluenes
reportedly dlmerlze nonenzymatlcally to azoxy compounds (Fewson, 1981).
Complete degradation of the aromatic ring of trinitrotoluene to carbon
dioxide does not occur, possibly because of the formation of these azoxy
dlmers, which are thought to resist degradation. The formation of the
degradation-resistant polyamldes described earlier may also be responsible
(Fewson, 1981).
2.2.5. 81oconcentrat1on. The BCF, useful In estimating the potential for
uptake 1n fish and aquatic organisms, can be calculated based on chemical
properties If 1t 1s not available experimentally. The linear regression
equation log BCF = 2.791 - 0.564 log S, where S 1s the water solubility 1n
ppm, can be used for this purpose (Bysshe, 1982). For trinitrotoluene, a
0240d -8- 11/01/89
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BCF of -45 can be obtained based on Us water solubility of 104-113 mg/l
at 20°C (Spanggord et al., 1983). This value suggests that bloaccumulatlon
In fish and aquatic organisms 1s not significant.
2.2.6. Adsorption. Pertinent data regarding the adsorption of trinitro-
toluene to sediment and suspended organic matter were not located 1n the
available literature cited In Appendix A. The potential for the strong
adsorption of trinitrotoluene to soil (Section 2.3.2.) suggests that adsorp-
tion to sediment may occur. However, Ryon et al. (1984) reported that
sorptlon to sediment was of minor Importance compared with Us photolysis
and blodegradatlon In natural waters. Under cloudy skies In winter, when
both photolysis and blodegradatlon are at their minimum, sorptlon to
sediment may become significant.
2.2.7. Volatilization. Using the estimation method of Hlne and Mookerjee
(1975), a Henry's Law constant of 3.30x10"' atm/m3-mol at 25°C can be
obtained for trinitrotoluene. An estimated volatilization half-life of
>16,000 days from a model river 1 m deep, flowing 1 m/sec, with a wind
velocity of 3 m/sec can be obtained using the above Henry's Law constant
(Thomas, 1982). This value suggests that volatilization of trinitrotoluene
from the water to the atmosphere will not be an Important process.
2.3. SOIL
2.3.1. M1crob1al Degradation. Trinitrotoluene underwent degradation In
an active compost system under aerobic conditions at 55°C. Although the
rate was not given, some material remained after 91 days. The metabolites
Identified were the same as those seen In the mlcroblal degradation of
trinitrotoluene 1n water (Kaplan and Kaplan, 1982a). Soil not previously
exposed to trinitrotoluene degraded trinitrotoluene, but at a slower rate
than that In experiments with water (Osmon and Klausmeler, 1973). Microbes
0240d -9- 11/01/89
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obtained from soil, compost or waste lagoon sediments and adapted to phenol
blodegraded trinitrotoluene at a slow but steady rate under aerobic condi-
tions, with maximum respiration reaching 21.2 times the endogenous level
(Chambers et al., 1963; Tabak et al.. 1964).
2.3.2. Adsorption. In a lyslmeter column study using four different
kinds of soil (clay, silt, loam and sandy loam), no trinitrotoluene was
found In water samples taken from the bottom of the column over a period of
6 months. None of the known metabolites of trinitrotoluene blodegradatlon
were found In the leachate. After the termination of the dynamic portion of
the experiment, trinitrotoluene was adsorbed to the soil of all four columns
(Kayser and Burllnson, 1988). These data suggest that trinitrotoluene and
Us metabolites have the potential for adsorbing strongly to soil.
2.3.3. Volatilization. Reported vapor pressures for trinitrotoluene 1n
the range 4.09x10'* to 8.02xlO'« mm Hg at 25°C (Jones, 1960; Pella,
1977) suggest that volatilization from dry soil to the atmosphere will not
be a significant process. Similarly, the estimated Henry's Law constant of
3.3x10"' atm/ma-mol (H1ne and Mookerjee, 1975) suggests also that this
process will not be significant 1n moist soil.
2.4. SUMMARY
The dominant fate process for trinitrotoluene Is expected to be destruc-
tion by direct photolysis. The half-life for the sunlight photolysis of
trinitrotoluene 1n pure water Is 14 hours over the continental United States
during the summer months (Mabey et al., 1983). The sunlight photolysis of
trinitrotoluene 1n distilled water at room temperature occurred at a rate
equal to 75X removal after 30 hours (Burllnson et al., 1973). The rate was
nearly pH-lndependent, Increased with the presence of humlc material, and
0240d -10- 11/01/89
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decreased 1n the presence of oxygen and other triplet quenchers (Mabey et
al., 1983). The biological degradation of trinitrotoluene 1n environmental
waters Is expected to occur under both aerobic and anaerobic conditions,
although the presence of additional nutrients may be required (Carpenter et
al., 1978; Osmon and Klausmeler, 1973; Chambers et al., 1963; Tabak et al.,
1964; Enzlnger, 1970; Spanggord et al., 1981, 1983; Fewson, 1981; Hoffsommer
et al., 1978). Products from both the aerobic and anaerobic degradation of
trinitrotoluene are believed to come from Initial reduction of one or more
of the nltro groups, which are then capable of undergoing nonenzymatlc reac-
tions, which may result In the formation of products that are degradation-
resistant. Complete degradation of trinitrotoluene to CO- has not been
observed. Neither hydrolysis nor volatilization to the atmosphere are
expected to be significant In water. Adsorption to sediment and suspended
organic matter may occur, although 1t Is not expected to be a significant
process (Ryon et al., 1984). B1oconcentrat1on In fish and aquatic organisms
Is not expected to be a significant process. If released to soil, mlcroblal
degradation Is expected to occur (Kaplan and Kaplan, 1982a; Chambers et al.,
1963; Tabak et al., 1964; Osmon and Klausmeler, 1973). The potential for
strong adsorption and, thus, low mobility In soil has been demonstrated by
Kayser and Burllnson (1988). Volatilization from the soil surface to the
atmosphere 1s not expected to be significant. In the atmosphere, trinitro-
toluene Is expected to exist In both the vapor phase and the partlculate
form (Jones, 1960; Pella, 1977; Elsenrekh et al., 1981). No experimental
data on the direct photolytlc degradation of trinitrotoluene were located In
the available literature; however, this compound reportedly undergoes
light-Induced decomposition In the solid state (Burllnson et al., 1973).
0240d -11- 09/12/89
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Therefore, direct photolysis of gaseous and partlculate trinitrotoluene may
be a significant process 1n the atmosphere. No experimental data on the
physical removal of trinitrotoluene were located; however, H may be
deposited on the earth's surface by rain and partlculate settling (Ryon et
al., 1984).
0240d -12- 09/12/89
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3. EXPOSURE
3.1. WATER
Trinitrotoluene was detected 1n an on-slte waste lagoon and 1n the
wastewater at an unspecified Army ammunition plant at 1314 and 19 yg/l,
respectively (Jenkins et al., 1986). The wastewater from a modern, counter-
current, continuous flow manufacturing process contained trinitrotoluene at
a concentration ranging from 101-142.9 mg/i (Nay et al., 1972). It was
also found In 20.3X of the ether extracts of condensate water obtained from
trinitrotoluene production and purification at a concentration range of
0.10-3.40 mg/i (Spanggord et al., 1982).
Trinitrotoluene was detected In shallow groundwaters obtained near
disposal beds on a Naval ammunition depot In Colorado at a maximum concen-
tratlon of 620 yg/l (Perelra et al., 1979). Also Identified 1n the
groundwater were known metabolites of the microblal degradation of trinitro-
toluene: 4-am1no-2,6-d1n1trotoluene and 2-am1no-4,6-dln1trotoluene.
Trinitrotoluene was also Identified 1n groundwater wells downgradlent from
the Cornhusker Army Ammunition Plant, Nebraska (Spaldlng and Fulton, 1988).
Trinitrotoluene was not found at two sites, one 200 miles off the coast
of Florida and the other 45 miles west of San Francisco, where old liberty
ships loaded with antiquated munitions were scuttled at sea (detection limit
2 ppt) (Hoffsommer et al., 1972; Hoffsommer and Rosen, 1972).
3.2. FOOD
Pertinent data regarding exposure to trinitrotoluene by Ingestlon of
food were not located 1n the available literature dted In Appendix A.
0240d -13- 11/01/89
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3.3. INHALATION
Little data on the concentration of trinitrotoluene In the atmosphere
were located In the available literature cited 1n Appendix A. The atmo-
spheric concentration of trinitrotoluene at an explosives manufacturing
plant in the United Kingdom ranged from <0.01-2.53 mg/m3; however, It was
not clear whether this was a measurement of trinitrotoluene vapor or
partlculates. Metabolites of trinitrotoluene excreted In urine were
determined In exposed workers. There was no correlation between urinary
excretion and atmospheric levels of trinitrotoluene. Oral uptake was
considered to be negligible. The levels excreted 1n urine were higher than
the theoretical maximum uptake by Inhalation. Because urine Is not the only
route of excretion, uptake of trinitrotoluene must occur, to a large extent,
by routes other than or-al and Inhalation. The authors concluded that
occupational exposure to trinitrotoluene by Inhalation 1s not expected to be
significant when compared with dermal exposure (Woollen et al., 1986).
3.4. DERMAL
Trinitrotoluene exposure by workers Involved 1n the manufacture of
explosives Is thought to occur predominantly through dermal contact (Woollen
et al., 1986). Extensive tests of dermal uptake, however, have not been
reported In the available literature.
3.5. OTHER
Trinitrotoluene was found In the urine of munitions workers 1n Israel at
a concentration of 11-278 ng/mi. Also found In this study were the
products of trinitrotoluene metabolism (Y1non and Hwang, 1986).
3.6. SUMMARY
Limited data on exposure to trinitrotoluene were located 1n the avail-
able literature cited 1n Appendix A. Dermal exposure for those working In
0240d -14- 04/06/90
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areas related to the production of armaments 1s reportedly more significant
than exposure by Inhalation (Woollen et al., 1986). Trinitrotoluene was
detected In groundwater and surface water samples near sites of Us produc-
tion or manipulation (Jenkins et al., 1986; Nay et al., 1972; Spanggord et
al., 1982; Perelra et al., 1979; Spaldlng and Fulton, 1988). Thus, the
potential for Ingestlon of this compound exists for a small group of the
population, although quantitative estimations of human exposure for the
group cannot cannot be made because of the lack of drinking water and food
monitoring data.
0240d -15- 04/06/90
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4. ENVIRONMENTAL TOXICOLOGY
4.1. AQUATIC TOXICOLOGY
4.1.1. Acute Toxic Effects on Fauna. A series of tests on the acute
toxlclty of 2,4,6-trlnKrotoluene to freshwater Invertebrates and fish was
performed by Liu et al. (1983). Static 48-hour assays In which members of
the test species (1n groups of 10 or 20) were exposed to five nominal
concentrations of 2,4,6-tMnltrotoluene and a control were done on Inverte-
brates. The 48-hour LC.. values recorded were 5.2 mg/l In the ollgo-
chaete worm, Lumbrlculus varleqatus. 6.5 mg/l In the scud, Hyalella
-i
azteca. 11.7 mg/l 1n the water flea, Daphnla magna. and 27.0 In the midge,
Tanytarsus dlsslmllls. Static 96-hour tests were done on fish using six
nominal concentrations (Including control), group sizes of 10 and duplicate
tests. All fish tested were Juveniles and all tests were conducted at 20°C
except for those using trout, which were conducted at 12°C. The 96-hour
LC.- values were 0.8-1.5 mg/l In rainbow trout, Salmo qalrdnerll. 2.4
mg/l In channel catfish, Ictalurus punctatus, 2.6-3.4 mg/l In blueglll
sunflsh, Lepomls macrochlrus. and 2.9 mg/l In fathead minnows, Plmephales
promelas. In a separate series of nonreplIcated tests on the fathead
minnow, It was found that pH had a minimal effect on 2,4,6-trlnltrotoluene
toxlclty, with the 96-hour LC.. Increasing from 1.2 mg/l at pH=5-2.1
mg/l at pH=7 and 2.4 mg/l at pH=9.4. These authors also conducted flow-
through acute toxlclty tests using measured concentrations of 2,4,6-trl-
nltrotoluene. The tests were conducted In duplicate using 10 worms, 15
water fleas or 20 fish per replicate. The 96-hour LC5_ value for L.
varlegatus was >29.0 mg/l and the Incipient LC5Q was 13.9 mg/l and was
reached after 336 hours. The 96-hour and Incipient LC5Q values In Daphnla
were 1.2 and 0.19 mg/l (after 192 hours), respectively. Among fish, the
0240d -16- 09/12/89
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96-hour LC5Q values were slightly higher than In the static tests, with
values of 2.0 mg/l 1n rainbow trout, 3.3 mg/l In channel catfish, 2.5
mg/i 1n blueglll sunflsh and 3.7 mg/l 1n fathead minnows. The Incipient
LCcg values and times they were reached In these species were 1.9 mg/l
(240 hours), 1.6 mg/l (288 hours), 1.4 mg/l (312 hours) and 1.5 mg/l
(384 hours), respectively.
Pederson (1970) reported the results of acute toxlclty bloassays on
blueglll sunflsh, L_. macrochlrus. These were 96-hour static tests and" the
water was renewed every 24 hours. The tests were run at either 10 or 25°C
and 1n either soft (60 ppm as CaCO-) or hard water (180 ppm as CaCO»).
w 0
For each of the four test series, one group of 10 fish was exposed at each
measured concentration. The 96-hour LC5Q values varied from 2.3-2.8
mg/l. This range Is similar to the values reported by Liu et al. (1983).
The LC5Q values were significantly lower at 10°C (2.3 mg/P) than at 25°C
(2.7-2.8 mg/i), Indicating that 2,4,6-tr1n1trotoluene was more toxic at
the lower temperature. Toxldty was not affected by water hardness In this
study.
Fathead minnows, P_. promelas. were exposed to measured concentrations of
2,4,6-trlnKrotoluene ranging from 0.05-44.9 mg/l under flowthrough condi-
tions (Smock et al., 1976). One group of 15 fish was tested at each concen-
tration. The 96-hour LC5Q was 2.58+0.1 mg/i. This Is similar to the
result reported by L1u et al. (1983); the concentration below which no
deaths were reported was 1.78 mg/l. Behavioral responses before death
were also noted. Gasping at the surface was the Initial reaction by the
fish to 2,4,6-tr1n1trotoluene. This reaction was followed by lethargy, loss
of motor control (exhibited by the fish swimming jerkily at the surface,
rapidly opening and closing their gills) and finally, lethargic swimming,
0240d -17- 11/01/89
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responding only to tactile stimuli. The 96-hour EC5Q for a behavioral
response to 2,4,6-tr1n1trotoluene was 0.46+^0.1 mg/l. No behavioral
responses were seen at concentrations of <0.05 mg/l.
4.1.2. Chronic Effects on Fauna.
4.1.2.1. TOXICITY Bailey (1982) used a model ecosystem to study
the chronic toxlclty of 2,4,6-trlnltrotoluene to Invertebrates. Five chemi-
cal concentrations and a control, each tested In duplicate, were Included.
Each microcosm was started with the green alga, Selenastrum caprlcornutum.
at an Initial density of 10,000 to 15,000 cells/ml, the benthlc o 1-1 go-
i
chaete, L.. varleqatus. at an Initial population of 30 and the water fleal'D.
maqna. at an Initial population of 15. The study was continued for 21 days,
with young Daphnla periodically counted and removed. Measurement of
2,4,6-trlnltrotoluene concentrations revealed that the chemical was steadily
lost from the water In this static system and the rate of loss was propor-
tional to the Initial test concentration. The total number of Daphnla
produced was reduced at concentrations of >0.6 mg/l. This could result
from a direct effect of the chemical on daphnld reproduction, or alterna-
tively, the daphnld population may have been limited by decreased algal
density. The total number of worms was reduced at 5.6 mg/l and the length
of these worms was also reduced. No animals of either species survived
21-day exposure to >10 mg/l.
4.1.2.2. BIOACCUHULATION/BIOCONCENTRATION -- Exploratory 4-day static
bloconcentratlon studies using 0.5 mg/l of 14C-labeled 2,4,6-trlnltro-
toluene 1n DMSO were performed on Invertebrates and fish by L1u et al.
(1983). The 4-day BCFs were 202.0 1n the ollgochaete worm, L. varleqatus.
(50 tested) and 209.0 1n the water flea, 0_. maqna (100 tested). BCFs of
338.0 and 9.5 were reported In the viscera and muscles, respectively, of the
0240d -18- 09/12/89
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blueglll sunflsh, L.. macrochlrus {three tested). The difference between
viscera and muscle BCF Is unknown, but the authors speculate that 2,4,6-trl-
nUrotoluene Is metabolized mostly In the liver and that radioactivity In
the viscera was concentrated In the liver. This chemical did not bloconcen-
trate significantly In this preliminary study.
4.1.3. Effects on Flora.
4.1.3.1. TOXICITY The effect of 2,4,6-trlnltrotoluene on algal
growth was studied by Smock et al. (1976). One series of tests was con-
ducted using the green alga, S. capMcornutum. Algal cultures with an
Initial concentration of 103 cells/mi were exposed to 2,4,6-trlnltro-
*
toluene concentrations ranging from 1-9 mg/i (three replicates per concen-
tration) under static conditions for 17 days. Ce>l counts were made
throughout the experiment. Concentrations <3 mg/i had no effect on algal
growth when compared with untreated controls. Growth was Initially
Inhibited at concentrations of >5 mg/l. Although growth later recovered
and no difference from controls was noted at the end of the experiment,
chemical analysis showed that recovery coincided with transformation of
2,4,6-trlnltrotoluene to other compounds. A similar study was conducted
using the blue-green alga, M1crocyst1s aeruglnosa (Initial concentration -60
mg/i blomass). The results had a pattern similar to those described above
except that the NOEC was 15 mg/i and the LOEC was 25 mg/i. The
blue-green algae colonies exposed to 2,4,6-trlnltrotoluene produced gas
vacuoles and gelatinous sheaths. This led to the formation of mats on the
water surface. The degree of matting was proportional to the concentration
of 2,4,6-trlnltrotoluene (1t was not seen In controls) and may have been a
reaction to the toxic environment.
0240d -19- 11/01/89
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Static toxlclty Dloassays were performed on the green alga, S. caprl-
cornutufn. the blue-green algae, M. aeruqlnosa and Anabaena flos-aquae. and
the diatom, Navlcula pelUculosa. using 2,4,6-tr1n1trotoluene (L1u et al.,
1983). Initial algae concentrations 1n culture were 10* cells for S.
capMcornutum and 5x10* cells for the other species. Each species was
exposed to six nominal treatment levels (Including control) and three
replicates were used for each level. Temperature was maintained at 24°C for
the 14 days of the study. Population growth was significantly reduced In
the green and blue-green algae at 2,4,6-tr1n1trotoluene concentrations of
>4.1 mg/l. In the diatom, concentrations of >18.0 mg/i had this effect;
however, photolysis of 2,4,6-trlnltrotoluene occurred during this study and
these results cannot be considered reliable.
Bailey (1982) used a model ecosystem to study the toxlclty of 2,4,6-trl-
nltrotoluene to the green alga, £. caprkornutum. Five chemical concentra-
tions and a control, each tested In duplicate, were Included. Each micro-
cosm was started at an Initial algal density of 10,000-15,000 cells/ml.
Other species Included In the microcosm were the benthlc ollgochaete, L..
vaMegatus. and the water flea, 0. magna. The study was continued for 21
days. Measurement of 2,4,6-trlnltrotoluene concentrations revealed that the
chemical was steadily lost from the water In this static system and that the
rate of loss was related to the Initial test concentration. Algal density
decreased soon after exposure to 2,4,6-trlnltrotoluene at concentrations
>5.6 mg/i. Algal density later decreased In the 0, 0.6 and 1.0 mg/l
groups. This was attributed to growth of the daphnld population.
BMngmann and Kuhn (1978) reported the results of cell multiplication
Inhibition tests on the green alga, Scenedesmus quadrlcauda. and the
blue-green alga, H. aeruqlnosa. Test cultures were maintained for 8 days
0240d -20- 09/12/89
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following addition of 2,4,6-trlnltrotoluene at various concentrations. At
the end of the experiment, algal concentrations were measured turbldl-
metrlcally. The toxlclty threshold (lowest concentration of 2,4,6-trlnltro-
toluene that produced Inhibition of cell multiplication) was 1.6 mg/i In
S. quadrlcauda and 0.32 mg/i In M. aeruqlnosa. In a screening-type study,
a concentration of 8 ppm (mg/i) of trinitrotoluene (Isomer not specified)
was reported to kill 100X of the blue-green alga, M. aeruqlnosa In culture
(Initial concentration -10* cells/mi) (Fitzgerald et al., 1952).
One study was conducted on a flowering aquatic plant, the duckweed.
Lenina perpusllla (Schott and Worthley, 1974). Each test was started with
two fronds, and growth results were tabulated 11 days later. The plants
were exposed to 2,4,6-tMnltrotoluene concentrations ranging from 0.01-50
pp'm (mg/i). Two replicates were used at each concentration and the study
was repeated under both acidic (pH=6.3) and basic (pH=8.5) conditions. No
effect on growth was seen at concentrations <0.5 ppm, but growth (number of
fronds 1n colony) was depressed compared with controls at 1 ppm; higher
concentrations were lethal to the plants. The results were not affected by
pH of the dilution water.
4.1.3.2. BIOCONCENTRATION Liu et al. (1983) conducted an explora-
tory bloconcentratlon test using the green alga, S. caprlcornutum. A
concentration of 0.5 mg/i of 2,4,6-trlnltrotoluene In DMSO was added to a
culture containing 10* cells and maintained under static conditions for 4
days at 24°C. A 4-day BCF of 453 was reported. This preliminary result
does not Indicate significant bloconcentratlon of 2,4,6-trlnltrotoluene.
4.1.4. Effects on Bacteria. Concentrations of 2,4,6-trlnltrotoluene <100
mg/i had no effect on the results of a cell multiplication Inhibition test
conducted using Pseudomonas put Ida. The toxldty threshold for 2,4,6-trl-
nltrotoluene 1n this test was >100 mg/l (BMngmann and Kuhn, 1980).
0240d -21- 11/01/89
-------
4.2. TERRESTRIAL TOXICOLOGY
4.2.1. Effects on Fauna. Pertinent data regarding the effects of
exposure of terrestrial fauna to 2,4,6-trlnltrotoluene were not located 1n
the available literature cited 1n Appendix A.
4.2.2. Effects on Flora. Yellow nutsedge plants, Cyperus esculentus.
were exposed to 2,4,6-tMnltrotoluene under static conditions for 42 days
(Palazzo and Leggett, 1986). Four groups, consisting of three plants each,
were exposed at each concentration (0, 5, 10 and 20 mg/i). Solutions were
renewed after 21 days when measurements revealed loss of 2,4,6-trlnltro-
toluene from the test solutions. Plant growth was significantly reduced
compared with untreated controls at concentrations of >5 mg/i. Total
plant yields were reduced 54-74X 1n treated plants. Most affected were the
roots and leaves whose weights were reduced 95-97% and 51-74%, respectively.
The effects did not Increase with dose; however, no difference was seen
between effects at 5 and 20 mg/i. This was true even though .the concen-
tration of 2,4,6-trInHrotoluene (and metabolites) In various parts of the
plant Increased with exposure concentration.
4.3. FIELD STUDIES
Pertinent data regarding the effects of 2,4,6-trInHrotoluene on flora
and fauna In the field were not located In the available literature dted 1n
Appendix A.
4.4. AQUATIC RISK ASSESSMENT
The lack of pertinent data regarding the effects of exposure of aquatic
fauna and flora to 2,4,6-tr1n1trotoluene prevented the development of a
freshwater criterion by the method of U.S. EPA/OWRS (1986). Available data
are displayed In Figure 4-1. Additional data required for the development
of a freshwater criterion Include the results of acute 4-day assays with a
0240d -22- 09/12/89
-------
TEST TYPE
Family
#1
Chordate (Salmonid-f ish)
12
Chordate (wannwater fish)
#3
Chordate (fish or amphibian)
#4 7
Crustacean (pianktonic)
#5
Crustacean (benthic)
#6
Insectan
#7
non-Arthropod/ -Chordate
#8
New Insectan or phylum
representative
#9
Algae
#10
Vascular plant
GMAVa
(mg/L)
2.0
2.5
3.1
1.2
NA
27.0
29.0
NA
NA
NA
GMCVa
(mg/L)
NA
NA
NA
NA
NA
NA
NA
NA
3.9
0.71
BCFa
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
'NA « Not available
FIGURE 4-1
Organization Chart for Listing GHAVs, GMCVs and BCFs Required to Derive
Numerical Water Quality Criteria by the Method of U.S. EPA/OWRS (1986) for
the Protection of Freshwater Aquatic Life from Exposure to 2,4.6-TMn1tro-
toluene.
0240d -23- 09/12/89
-------
benthlc crustacean and an Insect or species from a phylum not previously
represented. The development of a freshwater criterion will also require
data from chronic toxldty tests with two species of fauna and at least one
bloconcentratlon study. The chronic and bloconcentratlon studies reported
above did not meet the standards necessary for. Inclusion In calculation of a
criterion.
Pertinent data regarding the effects of exposure of marine fauna and
flora to 2,4,6-trlnHrotoluene were not located In the available literature
dted In Appendix A. Acute studies with representatives from eight families
of marine fauna and at least three chronic studies and one bloconcentratlon
study with marine fauna and flora are needed to develop a saltwater
criterion by the method of U.S. EPA/OWRS (1986).
4.5. SUMMARY
Static LC5Q values for 2,4,6-tMnltrotoluene varied from 5.2-27.0
mg/l among Invertebrates and 0.8-3.4 mg/l among fish (L1u et al., 1983;
Pederson, 1970). In flowthrough tests, LC-- values among fish ranged from
2.0-3.7 mg/l (L1u et al., 1983; Smock et al., 1976). D. magna was the
most sensitive species In flowthrough tests, with a 96-hour LC-- of 1.2
mg/l and an Incipient LC of 0.19 mg/l at 192 hours (Liu et al.,
1983). In a 21-day static model ecosystem, the LOEC values for reduced
population size and Individual growth were 0.6 mg/l In Daphnla and 5.6
mg/l In the ollgochaete L_. varleqatus (Bailey, 1982). Decreased algal
density may have been responsible for the reduction In Daphnla population.
The population density of the green alga S. c'aprlcornutum fell Immediately
following exposure to 5.6 mg/l In this study. Algal density also
decreased In microcosms exposed to lower concentrations, but this was a
delayed effect that may have been due to growth of the Daphnla population.
0240d -24- 11/01/89
-------
Other studies on algae reported LOEC values of 4.1-5 mg/l In S. caprl-
cornutum. 1.6 mg/l In S. quadrlcauda. 0.32-25 mg/l In M. aeruqlnosa. 4.1
mg/l In A. flos-aquae and 18 mg/l 1n the diatom, N. pelllculosa
(Brlngmann and Kuhn, 1978; Fitzgerald et al., 1952; Liu et al., 1983; Smock
et al., 1976). Photolysis of 2,4,6-tr1n1trotoluene was reported In two of
these studies and may have affected the results In others. The LOEC for
reduced growth was 1 mg/l In an aquatic flowering plant, the duckweed, I.
perpusHla. and 5 mg/l In a terrestrial plant*, the yellow nutsedge, C.
esculentus (Palazzo and Leggett, 1986; Schott and Worthley, 1974). Concen-
trations <100 mg/l had no effect on cell multiplication In the bacterium,
P. 'putlda. Several studies Included assays designed to determine the
Influence of water quality variables upon the toxldty of 2,4,6-tr1n1tro-
toluene In various species. None reported more than slight changes
attributable to these variables (Liu et al., 1983; Pederson, 1970; Schott
and Worthley, 1974). Exploratory 4-day static bloconcentratlon studies
reported BCFs ranging from 202-453 In several representative Invertebrates,
fish and algae.
0240d -25- 09/12/89
-------
5. PHARHACOKINETICS
A comprehensive study regarding absorption, distribution, metabolism and
excretion of 2,4,6-trInltrotoluene 1n rats, mice, rabbits and dogs using
oral, dermal and Intratracheal routes of exposure was conducted (El-hawar1
et al., 1981).
Male and female Sprague-Dawley rats and Swiss albino mice received
single gavage doses of 100 mg/kg body weight l4C-tr1n1trotoluene (ring-
labeled) and male and female New Zealand rabbits and beagle dogs were given
a single gavage dose of 5 mg/kg bw of labeled trinitrotoluene. The compound
was dissolved In peanut oil. Urine and feces were collected separately.
After 24 hours, the animals were sacrificed; blood was collected and tissues
and organs were removed and analyzed for radioactivity.
Male and female rats and male mice received a single application of 50
mg/kg body weight of radioactive trinitrotoluene In peanut oil on a clipped
area of their backs. Male rabbits and dogs were treated with either 5 or 50
mg/kg trinitrotoluene In peanut oil. Precautions were taken to prevent the
animals from grooming their fur. Concurrent experiments were conducted with
animals treated orally with the same dose of trinitrotoluene. Urine and
feces were collected separately for 24 hours. Blood samples were taken from
the tall vein of rats at 4, 8 and 24 hours after dosing. After 24 hours,
the animals were sacrificed and organs and tissues were removed for analysis
of radioactivity. Skin, Including that from the site of application, was
not retained for analysis.
A dose of 50 mg/kg body weight of radioactive trinitrotoluene suspended
In 0.5X methylcellulose was administered orally or Intratracheally to anes-
thetized and tracheotomlzed male rats. The trinitrotoluene particle size
0240d -26- 09/12/89
-------
was 1-3 pm. Serial blood samples were taken from the femoral artery over
a period of 4 hours; the rats were then sacrificed for tissue sampling. In
addition, bladder urine was collected for radioactivity analysis. In an
additional group of rats, bile samples were collected from the cannulated
common bile duct at different times after dosing and analyzed for radio-
activity. Blood samples were also collected, and the rats were sacrificed
after 4 hours for tissue sampling.
5.1. ABSORPTION
According to El-hawar1 et al. (1981), trinitrotoluene was readily
absorbed In the treated animals after oral exposure. The latter can be
Inferred by the recovery of radioactivity In the urine 24 hours after
dosing. The amount of radioactivity, expressed as percentage of the dose,
recovered In the urine Is presented In Table 5-1. The rate of absorption
was estimated only In rats. Following dermal exposure to a 50 mg/kg dose of
trinitrotoluene In this species, the radioactivity In the blood Increased
with time until >24 hours after dosing. In contrast, a 50 mg/kg oral dose
produced a peak of radioactivity In the blood at 8 hours. Although the
extent of oral absorption can only be approximated since the extent of
biliary excretion and enterohepatlc circulation was not studied, several
generalizations regarding absorption can be made based on urinary excretion:
there Is more trinitrotoluene absorbed after oral dosing than after dermal
administration; dogs and rabbits appear to absorb more trinitrotoluene after
oral administration than rats and mice; In decreasing order, dermal absorp-
tion Is greater In rabbits than In mice, rats and dogs. Based on blood
levels of radioactivity, oral absorption of trinitrotoluene by rats was
greater after 4 hours when the compound was suspended In methylcellulose
than when It was dissolved In peanut oil. Based on urinary excretion and
0240d -27- 11/01/89
-------
o
ro
o
o.
CO
I
TABLE 5-1
Excretion of Radioactivity 24 Hours After Administration of "C-Tr1n1trotoluenea»b»c
Species
Rat
Rat
Rat
Rat
Rat
Mice
Mice
Mice
Rabbit
Route
oral
oral
orald
dermal
1ntratracheald
oral
oral
dermal
oral
Sex
M
F
M
F
M
M
F
F
M
F
M
M
F
F
M
F
M
M
M
Dose/Vehicle
(mg/kg)
50/peanut oil
50/penut oil
100/peanut oil
100/peanut oil
50/methylcellulose
50/methylcel lulose
50/methylcel lulose
50/methylcellulose
50/peanut oil
50/peanut oil
50/methylcellulose
50/methylcellulose
50/methylcel lulose
50/methylcellulose
100/peanut oil
100/peanut oil
50/peanut oil
50/peanut oil
5/peanut oil
Ur
59
42
52
64
14
10
10
8
17
14
19
17
13
12
41
42
59
22
68
Ine
.5
.5
.7
.5
.6
.7
.0
.4
.4
.6
.3
.5
.2
.7
.9
.8
.1
.7
.1
Feces
10.7
2.1
8.1
2.1
NO
NO
NO
NO
1.3
2.5
NO
NO
NO
NO
22.0
8.9
24.1
14.2
5.4
Gastrointestinal
Tract and Contents
20
35
39
33
73
68
79
64
v ' '
3
6
18
1
12
2
13
7
10
3
19
.2
.3
.8
.9
.7
.3
.0
.2
.1
.4
.2
.8
.1
.9
.4
.4
.2
.6
.7
Bile
NO
NO
NO
NO
N0e
11. 6*
N0e
9.7^
NO
NO
NDe
19. 8f
N0e
14. 5f
NO
NO
NO
NO
NO
Recovery
92
81
91
102
93
95
97
91
22
24
45
47
40
45
80
60
94
41
95
.9
.3
.6
.4
.3
.5
.9
.2
.8
.9
.6
.1
.2
.0
.0
.4
.4
.7
.6
rsj
\
00
-------
TABLE 5-1 (cont.)
o
Q.
UJ
I
Species
Rabbit
Rabbit
Rabbit
Dog
Dog
Dog
Dog
Route
dermal
oral
dermal
oral
dermal
oral
dermal
Sex
H
H
H
H
M
M
H
Dose/Vehicle
(mg/kg)
5/peanut oil
50/peanut oil
50/peanut oil
5/peanut oil
5/peanut oil
50/peanut oil
50/peanut oil
Urine
52.8
74.3
47.2
70.5
11.7
61.0
11.8
Feces Gastrointestinal Bile
Tract and Contents
7.8
5.1
2.8
8.9
1.7
22.2
0.8
5.7
22.7
5.8
14.6
1.6
1.7
1.7
ND
NO
ND
ND
ND
ND
ND
Recovery
68.3
103.7
56.9
99.4
16.8
94.2
15.9
aSource: El-hawarl et al., 1981
bMean values of 3-6 rats, 6-8 mice. 2-4 rabbits and 1-3 dogs
Expressed as percent of administered dose
dSamples were collected 4 hours after dosing
eNo bile cannulated
ffJIle cannulated
S NO = Not determined
CO
VO
-------
amounts In the gastrointestinal tract and contents (see Table 5-1), intra-
tracheal Instillation of trinitrotoluene (conducted with only rats) resulted
In faster and greater absorption than after oral administration. In rabbits
and dogs, the extent of oral or dermal absorption seemed Independent of the
amount administered over a dose ranging from 5-50 mg/kg.
5.2. DISTRIBUTION
The distribution of radioactivity In tissues of the rat and rabbit
following different routes of administration of radioactive trinitrotoluene
Is shown In Table 5-2. In rats, the distribution of radioactivity was
similar after both oral and dermal administration of trinitrotoluene In
peanut oil. Intratrachael Instillation of a dose of 50 mg/kg resulted In
high accumulation of radioactivity at 4 hours In the liver, kidney, lung and
fat and (not shown In Table 5-2) blood and the gastrointestinal tract.
Levels of radioactivity In lungs and fat were markedly higher at 4 hours
than at 24 hours. In male rabbits, radioactivity In blood and residual bile
was higher after oral administration of a dose of 50 mg/kg of trinitro-
toluene (not shown )n Table 5-2). Increasing the oral or dermal dose by
10-fold resulted 1n a similar distribution as with the lower dose.
In other data (El-hawar1 et al., 1981), radioactivity In the blood,
liver, kidney, spleen, muscle and residual bile of dogs was higher after
oral administration of a 5 mg/kg dose of trinitrotoluene than after dermal
application of the same dose. The radioactivity In fat was higher after
dermal dosing. The distribution pattern of radioactivity after a dose of 50
mg/kg was similar to that seen after dosing with 5 mg/kg. Oral and dermal
administration of trinitrotoluene to rabbits and dogs resulted 1n greater
levels of radioactivity In the residual bile than 1n liver and blood.
0240d -30- 11/01/89
-------
O
a.
TABLE 5-2
Tissue Distribution of Radioactivity 24 Hours After Administration
of 14C-Tr1n1trotoluene Using Ratsa.b-c
I
CJ
ro
oo
Tissue
Liver
Kidney
Lungs
Spleen
Brain
Muscle
Fat
Sex
M
F
H
H
F
H
F
H
F
H
F
H
F
Oral 100 mg/kg
1n Peanut Oil
10.7
13.9
3.5
2.6
0.3
0.4
1.8
4.7
0.2
0.2
0.8
2.2
NO
NO
Oral 50 mg/kg
In Peanut Oil
7.3
5.5
5.8
4.5
2.1
2.1
1.0
1.0
0.6
0.5
0.9
0.7
1.1
0.8
Dermal 50 mg/kg
In Peanut Oil
2.8
3.1
3.1
4.0
1.4
1.7
0.6
0.5
0.9
1.2
0.6
1.1
2.4
3.8
Orald
50 mg/kg
1n Methyl-
cellulose
12.2
9.6
11.7
19.1
44.0
21.4
3.4
2.0
4.4
9.4
2.4
7.0
30.8
96.3
UHrad
Tracheal
50 mg/kg In
Methylcellulose
13.5
14.3
17.5
23.2
35.7
23.6
3.2
5.8
6.5
16.2
4.9
11.3
82.4
154.7
aSource: El-Hawar1 et al., 1981
bMean values of three to six rats
cvg/g tissue
dT1ssue samples were collected 4 hours after dosing.
NO - Not determined
-------
TABLE 5-3
Tissue Distribution of Radioactivity 24 Hours After Administration
of l4C-Tr1n1trotoluene Using Rabb1tsa»D'c
Tissue
Liver
Kidney
Lungs
Spleen
Brain
Muscle
Fat
Sex
M
F
M
M .
F
M
F
M
F
M
F
M
F
Oral 5 mg/kg
In Peanut 011
1.5
1.7
0.5
0.9
1.7
3.8
0.2
0.3
0.09
0.1
0.1
0.2
0.1
NO
Dermal 5 mg/kg
1n Peanut 011
1.0
NO
0.6
NO
0.6
NO
0.1
NO
0.09
NO
0.1
NO
0.2
NO
Oral 50 mg/kg
1n Peanut 011
8.7
NO
3.7
NO
2.4
NO
1.2
NO
0.5
NO
0.7
NO
1.8
NO
Dermal 50 mg/kg
1n Peanut 011
7.3
NO
6.9
NO
4.2
NO
1.0
NO
0.5
NO
0.6
NO
2.8
NO
aSource: El-Hawarl et al., 1981
bMean values of two to four rabbits
cyg/g tissue
dT1ssue samples were collected 4 hours after dosing.
NO = Not determined
0240d
-32-
04/23/90
-------
TABLE 5-4
Tissue Distribution of Radioactivity 24 Hours After Oral
Administration of l4C-Tr1n1trotoluene Using Dogsa«b»c
Tissue
Liver
Kidney
Lungs
Spleen
Brain
Muscle
Fat
Sex
H
F
M
F
M
F
M
F
H
F
M
F
M
F
5 mg/kg 1n
Peanut 011
4.0
2.6
1.1
1.6
0.7
1.5
1.0
1.3
0.3
0.4
0.2
0.3
NO
NO
50 mg/kg In
Peanut 011
22.6
NO
9.9
NO
8.7
NO
19.8
NO
2.2
NO
1.6
NO
5.2
NO
aSource: El-HawaM et al.. 1981
bHean values of one to three dogs
cyg/g tissue
NO = Not determined
0240d
-33-
04/23/90
-------
In the four animal species tested, tissue/blood concentration ratios 24
hours after oral or dermal administration of trinitrotoluene were >1.0 In
liver, kidney, lung and occasionally spleen and <1 In muscle and brain. Fat
tissue had tissue/blood ratios <1.0 after oral dosing and >1.0 after dermal
treatment. The tissues accumulated more radioactivity after Intratracheal
Instillation than after oral dosing.
5.3. METABOLISM
Analysis of trinitrotoluene metabolites 1n urine and bile was conducted
using TLC. Because of the large number of metabolites Identified, detailed
quantitative determinations were not attempted. A schematic presentation of
possible metabolic products and pathways Is shown 1n Figure 5-1.
The results of El-hawarl et al. (1981) Indicate that trinitrotoluene was
metabolized extensively In the four species studied, regardless of the route
of exposure. Large portions of the products were conjugated with glucuronlc
add. No conjugation with sulfuMc acid was detected. The urine from bile
duct-cannulated rats contained lesser amounts of glucuronldes than urine
from noncannulated rats. Most of the metabolic products found were nltro-
reductlon derivatives, such as the 2- and 4-hydroxylam1nes, the 2- and
4-monoam1nod1n1tro and 2,6- and 4,6-d1am1nomonon1tro compounds. There was
also evidence for oxidation of the methyl group. The parent compound,
trinitrotoluene, was present In small amounts 1n the urine of rats, mice and
dogs, but could not be demonstrated 1n rabbits.
Only quantitative differences were noted between the metabolic profiles
of rats, mice and dogs and between different routes of exposure. The urine
of rats contained large amounts of the 4,6-d1am1ne and lesser amounts of the
2,6-d1am1ne and either or both of the 2- or 6-monoam1nes. The 2- and
0240d -34- 04/06/90
-------
UXV)
(I) 4-Hydroxyl*aino-2.6-
-------
4-hydroxylam1nes were found In small quantities. The formation of tMnltro-
benzyl alcohol was also postulated. No significant differences 1n metabolic
profiles were detected between male and female rats. Also, the 4-hour urine
contained more of the polar metabolites and more parent compound than did
24-hour urine. Mouse urine contained smaller quantities of the polar
metabolites and the dlamlnes and more of the monoamlnes and hydroxylamlnes.
It also contained considerable amounts of trlnUrobenzyl alcohol and
tr1n1trobenzo1c add. The metabolic profiles of dog urine contained
appreciable amounts of dlamlnes and monoamlnes and possibly trinltrobenzyl
alcohol and trlnKrobenzolc acid. Only trace amounts of the 4-hydroxyl-
amlne, the 2-hydroxylama1ne and some azoxytoluene were found. Rabbit urine
was unique In that the presence of larger quantities of monoamlnes and
hydroxylamlnes was revealed. Furthermore, 1t contained either or both of
the dlamlnes. trinltrobenzyl alcohol and trlnltrobenzolc add. The only
significant difference between urine profiles from orally and dermally dosed
rats was that the latter excreted larger amounts of the parent compound.
After 0-glucuron1dase hydrolysis of the urine from the different
spedes, the extractable radioactivity Increased considerably. No major
changes In the profiles were noticed. The amount of glucuronldes varied
among spedes, with the least amounts observed In the urine of mice. Urine
from dermally exposed animals contained smaller amounts of glucuronlde
conjugates than did urine from orally exposed animals. The bile was found
to contain considerable amounts of low molecular weight glucuronlde
conjugates.
5.4. EXCRETION
The comparative excretion of radioactivity In the urine, feces and bile
In the four spedes examined (El-hawar1 et al., 1981) Is presented 1n Table
0240d -36- 04/06/90
-------
5-1. No attempt was made to determine elimination of radioactivity through
the exhaled air; however, the Investigators reported that previous experi-
ments 1n the same laboratory showed that -0.1% of an oral dose 1s eliminated
by this route. No significant differences 1n excretion were noted between
males and females. As seen In Table 5-1, large percentages of the orally
administered doses were excreted 1n the urine. The urine of rats and mice
had a bright red color. This seems to correspond to a partial reduction
product of 2,4,6-trlnltrobenzyl alcohol. Rabbits excreted a larger amount
of radioactivity In the urine 24 hours after dermal exposure compared with
the other species tested.
Biliary excretion seemed to play an Important role In the elimination of
trinitrotoluene In the four species examined, as Indicated by levels of
radioactivity located 1n residual bile. B1le duct-cannulatlon experiments
1n rats Indicated that biliary and urinary excretion may be equivalent In
the elimination of trinitrotoluene.
5.5. SUMMARY
In mice, rats, dogs and rabbits, trinitrotoluene administered orally,
dermally or Intratracheally (only to rats) 1s readily absorbed, distributed,
metabolized and excreted In the urine and, to a lesser extent, 1n feces
(El-hawar1 et al., 1981). Generally, the rate of absorption by the three
routes tested was Intratracheal > oral > dermal. The extent of absorption
In the four species tested was not significantly different when trinitro-
toluene was administered by the oral route. After dermal exposure, however,
the highest absorption occurred 1n rabbits, followed by mice, rats and dogs.
Radioactivity was mainly distributed to the liver and kidney of the animals
after oral dosing, but fat contained appreciable amounts of radioactivity
following dermal treatment. Trinitrotoluene was extensively metabolized In
0240d -37- 04/06/90
-------
all species examined regardless of the route of administration. Identifica-
tion of products In the urine and bile showed that most metabolites were
nltroreductlon derivatives. Oxidation of the methyl group had also
occurred. Unchanged trinitrotoluene could not be Identified 1n the urine of
rabbits. The metabolic profiles of urine from rats, mice and dogs and
different routes of exposure differed only quantitatively and no significant
sex differences were observed. Urine of rabbits was unique because It
contained larger amounts of monoamlnes and hydroxylamlnes. Urinary and
biliary excretion appear to play nearly equivalent roles In the elimination
-"!
of trinitrotoluene.
0240d -38- 04/06/90
-------
6. EFFECTS
6.1. SYSTEMIC TOXICITY
6.1.1. Inhalation Exposure. Pertinent data regarding Inhalation exposure
to 2,4,6-tr1n1trotoluene were not located 1n the available literature dted
In Appendix A.
6.1.2. Oral Exposure.
6.1.2.1. SUBCHRONIC In a study conducted by Levlne et al. (1983),
which Is the basis for the currently verified oral R£0 (Chapter 7), beagle
dogs (6/sex/dose) were administered dally gelatin capsules contalng
trinitrotoluene (99.1% pure) at doses of 0 (controls), 0.5, 2.0, 8.0 or 32.0
mg/kg/day for 26 weeks. All animals received a blank gelatin capsule for 1
week before testing. Physical examinations, Including body weights and food
consumption monitoring, were conducted 1 day/week, 3 weeks before
trinitrotoluene dosing. A complete hematologlcal profile, clinical
chemistry analyses and urlnalyses were performed on samples collected
several times during the pretest and test periods. Ophthalmic examinations
and electrocardiograms were also performed on all test animals during the
pretest and test periods. All surviving animals were sacrificed and
necropsled during test week 27 following a 16- to 18-hour fast. All major
organs and tissues were fixed for microscopic examination.
Clinical signs of toxldty attributed to trinitrotoluene treatment
Included transient ataxla, darkening of the tongue and gums, evidence of
Jaundice 1n animals receiving the 32 mg/kg/day dose, orange/brown urine and
orange/red feces 1n the two highest dose groups. H1stolog1cal examination
of masses, developed 1n two females 1n the two highest dose groups, was
unremarkable. Two treatment-related deaths occurred In the 32 mg/kg/day
female group before week 17. Both animals showed signs of malnutrition.
0240d -39- 04/06/90
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Significant body weight reduction was reported at the 8 (males only) and 32
mg/kg/day dose levels. Food consumption was significantly reduced for
animals at the highest dose level during most of the treatment period.
Dose-related anemia (decreased hematocrlt, hemoglobin and erythrocyte
counts) developed In all trinitrotoluene-treated dogs starting at week 3 and
lasted throughout the testing period. These changes were significant at the
two highest dose levels. Methemoglob1nemla occurred at the 8 and 32
mg/kg/day dose levels. Physiologic compensatory responses to anemia at
these doses Included retlculocytosls, mlcrocytosls and Increased numbers of
nucleated RBCs.
Clinical chemistry effects Included significant Increases 1n serum
globulin levels (at all doses) and serum LOH (In males and possibly females
1n the 32 mg/kg group) and decreases In SGPT In both sexes at all doses.
Total and direct blUrubln levels were elevated at the highest dose tested
In males and females and males only, respectively. Urinary blllrubln levels
were significantly Increased at the two highest dose levels from test week
17 and thereafter. Also, trace levels of uroblllnogen were seen during this
time In the two highest dosage groups. Opthalmologlc and electrocardlo-
graphlc testing revealed no conclusive effects attributable to trinitro-
toluene treatment.
Male (8 and 32 mg/kg/day) and female (32 mg/kg/day) dogs had significant
(p<0.05) Increases 1n relative and absolute liver weight. A slight but
statistically significant (p<0.05) Increase In relative kidney weights was
observed for high-dose females but not males. Relative and absolute In-
creases (p<0.05) 1n spleen weight occurred 1n females at 8 and 32 mg/kg/day,
but only at the latter dose In males. Hepatocytlc cloudy swelling and
hepatocytomegaly were present In all trinitrotoluene-treated groups;
0240d -40- 04/06/90
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however, the Incidence and severity of the lesion was dose-related. Micro-
scopic evidence of cirrhosis, observed only 1n treated animals, was reported
In one male at the 8 mg/kg/day dose level, six males at the 32 mg/kg/day
dose level and one female at the latter dose level. One female dog at the 2
mg/kg/day dose level and all animals at the 8 and 32 mg/kg/day dose levels
showed hemoslderosls In hlstocytes of the liver. The lesion was not
reported 1n controls. In addition, enlargement of the spleen with marked to
severe generalized congestion was attributed to trinitrotoluene treatment,
particularly at the two highest dose levels tested. None of the microscopic
lesions described were observed 1n the females necropsled before termina-
tion of the study. No NOAEL was Identified In this study and the dose level
of 0.5 mg/kg/day was Identified as a LOAEL for liver effects.
Levlne et al. (1984a) also conducted a subchronlc feeding study In rats.
In this study, F344 rats (10/sex/group) were fed a commercial diet contain-
ing trinitrotoluene (99.1% pure) at levels providing doses of 1, 5, 25, 125
or 300 mg/kg/day for 13 weeks. A group of 30 rats/sex served as untreated
controls. Test animals were monitored dally for lexicological signs.
Physical examinations, Including body weights and palpations for masses,
were performed weekly. Clinical biochemistry and hematologlcal tests were
performed on all survivors on week 13. Gross necropsy and hlstopathologlcal
examinations were conducted on all animals. Lethargy and ataxla were
observed In some animals receiving doses of >125 mg/kg/day throughout the
testing period. The Investigators attributed two deaths at the 300
mg/kg/day dose level on week 13 to severe anemia. Doses of >125 mg/kg/day
resulted 1n decreased food Intake with concommltant reduction 1n body weight
gain. Dose-related anemia (decreased hematocrlt, hemoglobin and erythrocyte
counts) was observed in treated rats. These changes were statistically
0240d -41- 04/06/90
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significant (p<0.05) 1n males at >25 mg/kg/day, but only at >125 mg/kg/day
1n females. Methemoglob1nem1a occurred In both males and females admin-
istered doses of 300 mg/kg/day. Elevated liver weights were reported In
male and female rats >25 mg/kg/day. Serum cholesterol levels and relative
spleen weights Increased significantly 1n both males and females at tri-
nitrotoluene dose levels >125 mg/kg/day. Dose-dependent congestive lesions
were observed 1n the spleen, whereas hyperplasla was noticed In the liver of
animals receiving doses >125 mg/kg/day. Degenerative lesions 1n tracts of
the cerebellar folia were seen at 300 mg/kg/day. H1stolog1cal examination
of the testes revealed dose-related degeneration of the germinal epithelium
lining the seminiferous tubules of males at 125 and 300 mg/kg/day. The
Investigators concluded that the liver, testes and blood are the main
targets of trinitrotoluene toxlclty and that the splenic lesions were
secondary to the hemolytlc effect.
Dllley et al. (1982) examined the toxldty of trinitrotoluene 1n sub-
chronic studies In mice, rats and dogs. Beagle dogs (5/sex/group) received
dally gelatin capsules containing equal weights of trinitrotoluene (>99%
pure) and lactose, which provided doses of 0 (control), 0.2, 2.0 or 20
mg/kg/day of trinitrotoluene. One male and one female were treated for 1
week and sacrificed. A second male and female were treated for the same
period and then allowed to recover for 4 weeks. The remaining dogs were
treated for 13 weeks and then two males and two females from each group were
sacrificed. The remaining dogs were allowed to recover for 4 weeks and then
sacrificed. Sprague-Oawley rats (20/sex/group) were fed trinitrotoluene 1n
a commercial diet at levels of 0 (control), 0.002, 0.01, 0.05 or 0.25%.
Based on food consumption and body weight data, the Investigators estimated
that the Intake of trinitrotoluene by treated rats was 1.40, 6.97, 34.7 and
160 mg/kg/day for males, and 1.45, 7.41, 36.4 and 164 mg/kg/day for females.
0240d -42- 04/06/30
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Five rats/sex/group were treated for 4 weeks and then sacrificed; 5/sex/
group were treated for 13 weeks and sacrificed; the remaining 5/sex/group
were treated for 13 weeks and were then allowed to recover for 4 weeks, at
which time they were sacrificed. The mice (Swiss-Webster strain) were fed a
commercial diet contalng trinitrotoluene at levels of 0 (control), 0.001,
0.005, 0.025 or 0.125%. Based on food consumption and body weight data, the
Investigators estimated that the Intake of trinitrotoluene was 1.56, 7.46,
35.7 or 193 mg/kg/day for males, and 1.57, 8.06, 37.8 or 188 mg/kg/day for
females. The group sizes, group compositions and treatment schedule were
the same as for rats. All animals were observed at least once dally and
weighed once weekly. Food consumption was recorded 5 days/week for dogs and
once weekly for rodents. Blood samples were taken periodically from dogs,
but only at terminal sacrifice from mice and rats. Hematologlcal parameters
were determined 1n the three species; clinical chemistry tests were
performed In dogs and rats only. Bladder urine was collected from dogs,
mice and rats at the time of sacrifice. All major organs and tissues were
examined grossly and then fixed for microscopic examination.
The resulting changes 1n hematologlcal parameters and In gross and
microscopical appearance of the spleen are suggestive of the development of
hemolytlc anemia at the highest dose used. Because the number of dogs used
was small, the significance of the results Is questionable but compatible
with the Levlne et al. (1983) study upon which the RfD was based.
Red-colored urine at the 0.05 and 0.25% levels was the only sign of
toxldty displayed by rats during treatment. However, urine color returned
to normal after treatment was discontinued. Hematologic changes seen In
rats Included low RBC counts, hemoglobin and hematocMt and Increased mean
corpuscular volume and leukocyte counts, particularly at the 0.25% dose
0240d -43- 04/06/90
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level. Other effects 1n rats were confined to the 0.25% group. A signifi-
cant (p<0.05) reduction 1n food consumption was seen, but food consumption
Increased when treatment was discontinued. Body weight gain was signifi-
cantly (p<0.01) reduced but recovered when treatment ceased. Spleen weight
and the spleen/brain weight ratios (both sexes) were significantly
Increased, and absolute and relative testes weight were significantly
decreased. At weeks 4 and 13, rats had a significant (p<0.01) Increase 1n
serum cholesterol. Also at week 13, SGPT and serum Iron levels were
markedly (p<0.01) reduced 1n males but not 1n females. Hemoslderosls of the
spleen (both sexes) and testlcular atrophy (males) accompanied by
hyperplasla were seen 1n animals sacrificed Immediately after treatment.
Hales allowed to recover for 4 weeks showed atrophy of the ep1d1dym1s.
Mice also had a red coloration in their urine, which disappeared after
cessation of trinitrotoluene treatment. No other overt signs of toxldty
attributable to treatment were noticed. Food consumption and body weight
decreased Initially In the groups at the highest dose and remained slightly
low for females given the 0.125% trinitrotoluene diet. Changes In body
weight were sporadic and Inconsistent. Treatment with trinitrotoluene
Induced an Increase 1n absolute and relative spleen weight, but a dose-
response relationship was not always obvious. Increased liver weight and
occasional necrosis was also noticed in high-group male mice sacrificed at
week 17. M1ld hematologlcal changes Indicative of hemolytlc anemia were
seen 1n mice at the 0.125% dose level. No hlstopathologlcal signs were
noticed In mice sacrificed after 4 weeks of treatment, with or without a
recovery period. Hemoslderosls of the spleen was observed 1n mice (3/5
males, 5/5 females) after 13 weeks of treatment at the 0.125% trinitro-
toluene level and in 1/5 male and 4/5 females at the 0.025% level at week 17.
0240d -44- 04/23/90
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Martin (1974) conducted a study using cynomolgus monkeys (3/sex/group)
In which trinitrotoluene (purity not reported) suspended In methylcellulose
was administered by gavage at doses of 0 (solvent alone), 0.02, 0.1 or 1.0
mg/kg/day for 90 days. Hematology, clinical chemistry tests, urlnalysls and
liver function tests revealed no alterations attributable to trinitrotoluene
treatment. Gross lesions, observed only with the two higher doses, Included
two cases of subcapsular renal hemorrhage and one monkey with mucosal
reddening and focal thickening of the large Intestine. H1stolog1cal exami-
nation revealed some Increases In the numbers of necrotlc megakaryocytes In
'?'
bone marrow and Increased amounts of 1ron-pos1t1vel material In the liver.
According to the Investigators, the significance of the hlstologkal
findings 1s uncertain.
Hart (1974) also conducted a study 1n which beagle dogs (3/sex/dose)
were fed a commercial diet containing trinitrotoluene (purity not reported)
for 90 days. This diet provided trinitrotoluene doses of 0 (control), 0.02,
i
0.1 or 1.0 mg/kg/day. Hematology, clinical chemistry tests, urlnalysls and
gross and microscopic appearance of organs and tissues were not affected by
administration of trinitrotoluene. Temporary episodes of emesls occurred
but tolerance appeared to develop.
6.1.2.2. CHRONIC Furedl et al. (1984a) evaluated the chronic
toxldty of trinitrotoluene 1n rats (Section 6.2.2.). In this study, 6- to
7-week-old F344 rats (75/sex/dose) were administered trinitrotoluene (>99%
pure) mixed In a commercial diet for <24 months. According to the Investi-
gators, this diet provided doses of 0 (control), 0.4, 2, 10 or 50 mg/kg/day
of trinitrotoluene. Ten rats/sex/dose were sacrificed following 27 and 53
weeks on test and the remaining animals were sacrificed after 24 months of
treatment. All animals were observed once dally for pharmacological and
0240d -45- 04/06/90
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lexicological signs. Physical examinations were conducted weekly until test
week 13 and biweekly thereafter. Food consumption and body weights were
also monitored weekly until test week 13 and biweekly thereafter until
termination of the study. Ophthalmic examinations were performed before
testing commenced and during weeks 25, 51, 76 and 103. Complete hemato-
loglcal and clinical chemistry tests were performed on blood samples drawn
from the same 10 rats/sex/dose during weeks 14, 26, 52, 78 and 104. The
brain, gonads, heart, liver, kidneys, spleen, spinal cord, pituitary gland,
urinary bladder (females) and sternal bone marrow (females) from rats
receiving doses of 0.4, 2.0 and 10 mg/kg/day were examined microscopically.
Additional tissues and organs from control and 50 mg/kg/day dose groups were
also examined.
Administration of trinitrotoluene did not affect survival rate nor did
it induce signs of toxlclty other than an apparent Increase In ocular
discharge In high-dose males during the second year of the study. Doses of
trinitrotoluene >10 mg/kg/day Induced dose-related reduction In body weight
gain and In food consumption. A dose-related reduction In hematocrlt,
hemoglobin and RBC count was observed In males throughout the study and In
females during the first year at the 10 and 50 mg/kg/day dose level.
Compensatory responses to the anemic state were minimal. The only other
hematologlcal effects seen, considered related to trinitrotoluene-treatment,
were methemoglob1nem1a 1n males at 10 and 50 mg/kg/day and thrombocytosls In
male and female rats at 50 mg/kg/day during the second year of the study.
Clinical chemistry tests showed an increase 1n serum cholesterol In
males with doses >2.0 mg/kg/day and In females at 50 mg/kg/day. At week
104, females In the highest dose group had a decrease In serum trlglycerlde
levels; males showed hypertr1glycer1dem1a. In general, serum total protein,
albumin and globulin levels were Increased In male and female rats given
0240d -46- 04/06/90
-------
doses of trinitrotoluene of 50 mg/kg/day. In addition, high-dose rats had
slightly Increased BUN levels during the second year of the study. Other
changes 1n clinical chemistry parameters were sporadic and not considered
related to trinitrotoluene treatment.
Administration of trinitrotoluene did not Induce ophthalmologlc
abnormalities. Dose-related hepatomegaly and Increased kidney weights were
seen during Interim sacrifices at weeks 27, 53 and at the end of the study
period at dose levels of 10 and 50 mg/kg/day. Absolute spleen weight (both
sexes) was Increased at weeks 27 and 53 In animals administered 50
mg/kg/day. Relative heart weight Increased In females at weeks 27 and 53
and in ...-.h sexes at week 105 at trinitrotoluene dosage levels of 10 and 50
mg/kg/day. Treatment-related lesions were seen In the spleen and kidneys of
rats sacrificed after 27 and 53 weeks of treatment. The hlstologlcal
changes were seen primarily at doses >2.0 mg/kg/day. Increased pigmentation,
sinusoidal congestion and extramedullary hematopolesls were seen 1n the
spleen. Changes In the kidney Included hypertrophy of proximal convoluted
tubules. Increased pigmentation and chronic nephropathy.
After 24 months of treatment, lesions were present 1n the liver of
males, urinary bladder and bone marrow of females and the spleen and kidney
of both sexes. Male rats 1n the 10 and 50 mg/kg/day dose levels had a
dose-related Increased Incidence of hepatocellular hyperplasla associated
with pellosls and cystic degeneration. Urinary bladder lesions In females
Included hyperplasla of the mucosal epithelium at >10 mg/kg/day. A signifi-
cant Increase In Incidence of sternal bone marrow flbrosls was observed 1n
females with doses >2.0 mg/kg/day. The latter effect was also seen 1n males
at 50 mg/kg/day, but lower dose males were not examined. Based on the
occurrence of splenic, renal and bone marrow effects at doses >2.0
mg/kg/day, a NOEL of 0.4 mg/kg/day can be Identified from this study.
0240d -47- 04/06/90
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Furedl et al. (1984b) also investigated the chronic toxlclty of
trinitrotoluene In mice (Section 6.2.2.). In this study, B6C3F1 hybrid mice
(75/sex/dose) were administered trinitrotoluene (>98.8% pure) mixed In a
commercial diet for <24 months. According to the Investigators, this diet
provided doses of trinitrotoluene of 0 (control), 1.5, 10 or 70 mg/kg/day.
The protocol used and the endpolnts examined were Identical to the ones used
in the rat study {Furedl et al., 1984a). Trinitrotoluene administration did
not affect mortality rate. Reductions In body weight gain for both sexes
were seen in animals receiving 10 and 70 mg/kg/day, but this effect was
significant only at the highest dose tested. Sporadic and mild episodes of
anemia (reduced hematocrH, hemoglobin and RBC count) were observed 1n males
and females at 70 mg/kg/day. No compensatory responses were apparent.
Furthermore, splenic lesions Indicative of hemolytlc anemia were not
observed 1n this study. Hepatomegaly was present at 70 mg/kg/day, but
without hlstologlcal alterations. Weight changes In other organs were
described as sporadic and not supported hlstologlcally.
6.1.3. Other Relevant Information. Dllley et al. (1982) administered
single gavage doses of trinitrotoluene dissolved In corn oil to Swiss-
Webster mice and Sprague-Oawley rats. Oral LDrQs of 660 mg/kg 1n male and
female mice and 1320 and 795 mg/kg In male and female rats, respectively,
were reported. The mice and rats were observed for <14 days. Signs of
toxlclty Included Inactivity, development of tremors and mild convulsions
and death. Animals that survived convulsions were still alive after 14 days.
Levlne et al. (1984b) studied the acute effects of trinitrotoluene 1n
hybrid B6C3F1 mice (10/sex/dose). Trinitrotoluene (>99% pure) was mixed In
the diet to provide doses of 0, 0.3, 2, 14, 100 or 700 mg/kg/day for 28
days. Trinitrotoluene did not affect survival rate at any dose level.
0240d -48- 04/06/90
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Weight loss (700 mg/kg/day) and weight gain reduction (TOO mg/kg/day) were
the only clinical signs observed. Treatment-related morphologic alterations
(hemoslderosls) were seen In the spleens at 100 and 700 mg/kg/day. Further
toxic effects of trlnltrotolueune seen primarily at the 700 mg/kg/day dose
level were leukopenla, thrombocytosls, slight hepatomegaly, marginal
decrease In testes weight and Increased kidney weight. Organ weight
changes, however, were not accompanied by hlstologlcal alterations. A NOAEL
of 14 mg/kg/day was Identified.
Single IntraperHoneal Injections of 100 mg/kg of trinitrotoluene In
olive oil to adult male Wlstar rats caused damage In cerebral, hepatic and
renal blomembranes (Zlttlng et al., 1982). According to the Investigators,
Intracellular damage observed, predominantly In brain and kidney. Is
consistent with the formation of superoxldes in aerobic conditions during
n1troreductlon of trinitrotoluene.
Cases of human exposure to trinitrotoluene are numerous but Involved
multiple routes of exposure. Details regarding levels and exposure
durations were usually Incomplete, rendering these studies unsuitable for
quantitative risk assessment. A review of occupational exposure studies
(Hathaway, 1977) Indicated that workers exposed to air levels between 0.01
and 4.0 mg/m3 may develop skin Irritation, liver damage and anemia.
Morton et al. (1976) reported that workers In an ammunition plant producing
trinitrotoluene had significant Increases In serum LDH (p<0.005) and SGOT
(p<0.01) when the concentration of trinitrotoluene 1n the air Increased from
0.3 to 0.8 mg/ma In -30 days because of an Increase 1n the trinitrotoluene
production rate. Hemoglobin values were not significantly different from
values before the production Increase. When affected Individuals were
removed from exposure, H took 1-3 weeks for LDH values to return to normal
range. Goodwin (1972) reported a mean of 1.80 MacLagen units In the thymol
0240d -49- 04/06/90
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turbidity test In 1537 workers exposed to trinitrotoluene In an ammunition
shell loading plant. The author considered the test to be an Indication of
hepatic Irritation. Mean preemployment levels were 0.93 MacLagen units.
Levels of trinitrotoluene 1n the workroom atmosphere ranged from 0.2-4.7
mg/m3 with a mean of 2.38 mg/m3. Reportedly, 36 workers (2.0%) had
"classical symptoms of liver damage."
Cone (1944) reported that transient leukocytosls and moderate eoslno^
ph1!1a developed 1n 17 workers exposed to trinitrotoluene levels between 0.5
and 2.0 mg/m3 of trinitrotoluene In the air (duration not specified) as
compared with preexposure levels. Stewart et al. (1945) reported the case
of 62 employees of a munitions loading plant exposed to a presumed average
trinitrotoluene level between 0.3 and 1.3 mg/m3 for 4-11 weeks. Skin
exposure was Inferred by the appearance of skin rashes and 85% of the
Individuals had considerable reduction 1n their hemoglobin levels. RBC
counts decreased and blUrubln levels Increased. FMedlander et al. (1974)
reported that anemia developed In workers exposed to trinitrotoluene levels
between <0.03 and 4.0 mg/m3. No differences, however, were noticed In
clinical parameters between test Individuals and unexposed controls 1n a
follow-up study performed at the same facility after reducing the exposure
levels to 0.08-0.59 mg/m3 for 8 hours/day. Buck and Wilson (1975)
reported that In a case of 533 employees (865 controls) exposed to trinitro-
toluene levels ranging from <0.01-1.84 mg/m3 (only 12.2% were exposed to
>0.5 mg/m3), the only significant finding was an Inverse relationship
between levels of exposure and hemoglobin concentration. Harkonen et al.
(1983) reported that 6/12 workers exposed to trinitrotoluene levels In the
air between 0.14 and 0.58 mg/m3 for a mean duration of 6.8 years developed
equatorial lens opacities. There was no effect on visual acuity or visual
fields. Blood chemistry and hematologlcal tests were unremarkable. These
0240d -50- 04/06/90
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findings suggest that the eye may be the critical target organ for chronic
exposure to low levels of trinitrotoluene 1n the air.
6.2. CARCINOGENICITY
6.2.1. Inhalation. Pertinent data regarding the Inhalation cardnogen-
Idty of trinitrotoluene were not located 1n the available literature dted
1n Appendix A.
6.2.2. Oral. The carcinogenic properties of trinitrotoluene In rats were
studied by Furedl et al. (1984a). In this study, 6- to 7-week-old F344 rats
(75/sex/dose) were administered trinitrotoluene (>99% pure) mixed In a
commercial diet for <24 months. According to the Investigators, this diet
provided doses of 0 (control), 0.4, 2, 10 or 50 mg/kg/day of trinitro-
toluene. A complete description of the protocol and of the noncardnogenlc
effects was provided In Section 6.1.2.2. Observations regarding systemic
toxldty Indicated that the MTD had been achieved (U.S. EPA, 1988a). A
significant Increase 1n the combined Incidence of urinary bladder papHloma
and carcinoma was observed at terminal sacrifice In female rats administered
the 50 mg/kg/day dose of trinitrotoluene (Table 6-1). No such lesions were
noticed at Interim sacrifices after 6 or 12 months of treatment. The Inves-
tigators Indicated that the fact that Incidences of hepatocellular (male
rats) and renal and urinary bladder hyperplasla (female rats) also signifi-
cantly Increased at the 50 mg/kg/day dose level support the conclusion that
trinitrotoluene 1s carcinogenic to F344 rats under these experimental
conditions (Table 6-2). '
The carcinogenic potential of trinitrotoluene was also Investigated 1n
mice (Furedl et al., 1984b). In this study, 4- to 5-week-old B6C3F1 hybrid
mice (75/sex/dose) were administered trinitrotoluene (>98.8% pure) mixed 1n
a commercial diet for <24 months. According to the Investigators, this diet
provided doses of trinitrotoluene of 0 (control), 1.5, 10, or 70 mg/kg/day.
0240d -51- 04/23/90
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TABLE 6-1
Incidence of Urinary Bladder Tumors 1n Female F344 Rats fed Diets
Containing Trinitrotoluene (>99% pure) for 24 Months3
Doseb
(mg/kg/day)
0
0.4
2
10
50
Tumor Type
papllloma
carcinoma
combined
papllloma
carcinoma
combined
papllloma
carcinoma
combined
papllloma
carcinoma
combined
papll loma
carcinoma
combined
Incidence
0/54
0/54
0/54
0/54
0/54
0/54
0/55
0/55
0/55
1/55
0/55
1/55
5/55c
12/55"!
17/55d
aSource: Furedl et al.. 1984a
^Provided by Investigators
cp<0.05
Vo.oi
0240d
-52-
04/06/90
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TABLE 6-2
Incidence of Hyperplastlc Lesions )n F344 Rats Fed Diets Containing
Trinitrotoluene (>99X pure) for 24 months3
Dose0
(mg/kg/day)
0
0.4
2
10
50
Hepatocellulor Hyperplasla
(males)
6/26
7/22 '
6/20
16/14d
27/12d
Urinary Bladder Hyperplasla
(females)
0/37
0/40
0/40
2/46
12/47d
aSource: Furedl et al., 1984a
^Provided by Investigators
Cp<0.05
dP<0.01
0240d -53- 04/23/90
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The use of these doses Is based on the results of a range-finding study by
the same Investigators (Levlne et al., 1984b) 1n which an MTD between H and
100 mg/kg/day was estimated. The protocol used and the endpolnts examined
were Identical to the ones used In the rat study (Furedl et al., 1984a). A
complete description of the noncardnogenlc effects was provided In Section
6.1.2.2. Neoplastlc lesions observed after 6 and 12 months of treatment
were considered Incidental and not treatment-related. A significant
(p<0.05) Increase In the combined Incidence of leukemia/malignant lymphoma
In the spleen was observed In females at the 70 mg/kg/day dose level. The
Incidences were 9/45, 15/39, 17/37 and 21/33 1n the 0, 1.5. 10 and 70
mg/kg/day dietary levels, respectively. U.S. EPA (1988a), however,
concluded that these tumors were not chemical-related because.when malignant
lymphomas and lymphocytk leukemia 1n all tissues were combined rather than
considered separately, there was neither a significantly Increased Incidence
nor a significant trend.
Trinitrotoluene has not been scheduled for cardnogenlclty testing by
NTP (1989).
6.2.3. Other Relevant Information. The carcinogenic potential of
2,4,6-tr1n1trotoluene Is supported by the cardnogenlclty of the
structurally related 2,4- and 2,6-d1n1trotoluene. Both Isomers are
considered to be class 82 probable human carcinogens based upon the
existence of sufficient evidence of cardnogenlclty 1n two spedes of
animals (U.S. EPA, 1988c).
The possibility that one or more' of the metabolites of
2,4,6-trlnltrotoluene was carcinogenic was also considered. No Information
on the cardnogenlclty or mutagenldty of the proposed metabolites was found
1n the available literature but an examination of the structure of the
compounds reveals several that are likely to form DNA adducts. The
0240d -54- 04/23/90
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N-hydroxylated species (II), as well as amlnobenzene derivatives (III, IV,
XIII) seem likely to be mutagenlc and to have potential as carcinogens
(Figure 5-1). Many of the remaining compounds are highly polar. Those
compounds would be unlikely to form ONA adducts and would be rapidly
excreted (El-hawar1 et al., 1981).
6.3. MUTAGENICITY
Trinitrotoluene gave positive mutagenlc responses when tested by the
reverse mutation assay In several strains of Salmonella typhlmurlum In the
absence of activating systems (Table 6-3). In the presence of activating
systems, trinitrotoluene was nonmutagenlc (Won et al., 1976; Whong and
Edwards, 1984). Furthermore, the mutagenkHy of trinitrotoluene 1n S.
typhlmurlum was heavily dependent on the presence of nltroreductases (Whong
and Edwards, 1984). The latter Indicates that reduction of the nltro
groups, possibly to hydroxylamlno Intermediates, may be an essential step
for the In vitro mutagenlc activity. When tested In mammalian systems In
vivo, trinitrotoluene did not Induce chromosome damage In mice bone marrow
cells or unscheduled DNA synthesis In rat hepatocytes (Ashby et al., 1985).
However, trinitrotoluene was mutagenlc In mouse lymphoma cells 1n culture 1n
the absence of an activating system (Styles and Cross, 1983).
6.4. DEVELOPMENTAL TOXICITY
Pertinent data regarding the developmental effects of trinitrotoluene
were not located 1n the available literature dted 1n Appendix A.
6.5. OTHER REPRODUCTIVE EFFECTS
Pertinent data regarding other reproductive effects of trinitrotoluene
were not located In the available literature cited 1n Appendix A.
6.6. SUMMARY
Reported oral LD5Qs for trinitrotoluene administered by gavage were
660 mg/kg In male and female mice and 1320 and 795 mg/kg In male and female
0240d -55- 04/23/90
-------
TABLE 6-3
NutagenlcUy Testing of 2.4.6-TMnHrotoluene
0
o.
1
tn
1
04/23/90
Assay
MICROBIOLOGICAL ASSAY
Reverse mutation
Reverse mutation
Reverse mutation
Reverse mutation
MAMMALIAN SYSTEMS
Bone marrow
mlcronucleus assay
(chromosome damage)
ONA repair
(UOS)
Forward mutation
NR * Not reported; NA
Indicator
Organism
Salmonella
typhlmurlum
TA1535. TA100.
TA1S37. TA1S38,
TA9B
S. typhlmurlum
TA98. TA1538.
TA100
S. typhlmurlum
TA98
S. typhlmurlum
TA9B
male mice
(CBAxBalbC)Fl
male rats
Alderley Park
and/or F344
P388 mouse
lymphoma
cells
= not applicable;
Purity
commercial
grade
commercial
grade
explosive
grade
explosive
grade
>99.0
>99.0
NR
NC = no comment
Application
plate
Incorporation
plate
Incorporation
plate
Incorporation
plate
Incorporation
Intraperltoneal
gavage
cell culture
Concentration Activating Response Comment
or Oose System
11-176 nmol/ -S-9 * Mutagenldty was lost
plate In the presence of
activating system or when
a nltroreductase-def Iclent
strain was used
5-500 ng/plate »S-9 * NC
*
0.5-10 |ig/mt -S-9 » Frameshlft mutagen. No
mutagenldty was seen when
tested In cultures Inocu-
lated with base substitution
tester strains
0.5-10 rog/ml tS-9 - NC
40 mg/kg NA - Mice received a single
Intraperltoneal Injection
of test compound. Sampling
was done at 24. 48 and 72
hours after Injection
100-1000 mg/kg NA - Hepatocytes were Isolated
12 hours after dosing
0-1000 wg/mi -S-9 * In the presence of activat-
ing system TNT was not
mutagenlc
Reference
Uhong and
Edwards. 1984
Kaplan and
Kaplan. 1982a
Won et al..
1976
Won et al.,
1976
Ashby et al.,
1985
Ashby et al.,
1985
Styles and
Cross. 1983
-------
rats, respectively. LD50 data for other species were not available. Data
were not available regarding the toxldty of trinitrotoluene to animals by
Inhalation exposure.
Data regarding the toxldty of trinitrotoluene 1n humans Indicated that
workers exposed to air levels between 0.01 and 4.0 mg/m3 may develop skin
Irritation, liver damage and anemia (Hathaway, 1977; Morton et al., 1976).
Although there are numerous reports of occupational exposure to trinitro-
toluene, the duration and levels of exposure were usually not sufficiently
defined to permit use of these studies 1n risk assessment.
Trinitrotoluene yielded evidence of carclnogenlclty In a 24-month
dietary exposure study using rats (Furedl et al., 1984a). In that study,
female rats had a significantly Increased Incidence of ur'nary bladder
papniomas and carcinomas. Trinitrotoluene was not carcinogenic when tested
1n mice (Furedl et al., 1984b). Trinitrotoluene was mutagenlc In reverse
mutation assays In S. typhlmuMum In the absence of activating systems
(Kaplan and Kaplan, 1982b; Hhong and Edwards, 1984; Won et al., 1976). The
presence of activating systems reduced the mutagenlc potency. Trinitro-
toluene did not Induce DNA damage 1n mammalian test systems hi vivo (Ashby
et al., 1985).
Subchronlc studies using animals suggest that dogs are the most sensi-
tive species since a dose of 0.5 mg/kg/day for 26 weeks Induced signs of
anemia and liver alterations (Levlne et al., 1983). Increasing doses
Increased the severity of the effects. Death occurred with a dose of 32
mg/kg/day before week 17. In contrast, death from anemia occurred In rats
with doses of 300 mg/kg/day (-10 times higher than 1n dogs) administered for
13 weeks (Levlne et al., 1984a). Mice appeared to be less sensitive. A
dose of 190 mg/kg/day for 13 weeks produced liver effects, but 36 mg/kg/day
0240d -57- 04/23/90
-------
was without adverse effects; H 1s possible that doses between 36 and 190
mg/kg/day could have been toxic. Chronic studies have been performed only
on rats and mice. In rats, a dose of 2.0 mg/kg/day 1n the diet for 24
months caused kidney hypertrophy, spleen congestion and bone marrow flbrosls
1n females. A dose of 10 mg/kg/day Induced signs of anemia, changes In
organ weights and urinary bladder lesions In females. In contrast, a dose
of 10 mg/kg/day administered In the diet for 24 months to mice was without
adverse effects. In general, the most sensitive endpolnts f.or assessing
t,ox1colog1cal effects of trinitrotoluene seem to be the liver and elements
.'.
in the blood, such as the RBCs.
Data regarding the developmental and reproductive toxldty of
trlnHroluene were not available 1n the literature cited In Appendix A.
0240d -58- 04/23/90
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7. EXISTING GUIDELINES AND STANDARDS
7.1. HUMAN
The U.S. EPA (1988b) verified a chronic oral RfD of 5xlO"4 mg/kg/day
for 2,4,6-tr1n1troto1uene based on a LOAEL of 0.5 mg/kg/day for liver
effects In a 26-week feeding study using beagle dogs conducted by Levlne e't
al. (1983). An uncertainty factor of 1000 was used.
ACGIH (1988) recommended a TLV-TWA of 0.5 mg/m3 for trinitrotoluene.
This recommendation 1s based largely on the conclusions of Goodwin (1972)
and Morton et al. (1976) who reported liver damage and alterations 1n
biochemical parameters In workers at munitions plants (ACGIH, 1986).
Trinitrotoluene was also Identified as a methemoglobln Inducer (ACGIH,
1988). In addition, ACGIH (1988) Indicates that dermal contact may
contribute to overall exposure. OSHA (1989) established a PEL for trinitro-
toluene of 0.5 mg/m3 TWA.
7.2. AQUATIC
U.S. Army limits of 1 mg/i In potable water and 5 mg/i In waters
used by fish and wildlife were reported by Smock et al. (1976).
0240d -59- 04/23/90
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8. RISK ASSESSMENT
8.1. CARCINOGENICITY
8.1.1. Inhalation. Pertinent data regarding the Inhalation cardnogen-
1dty of 2,4,6-trln1trotoluene were not located In the available literature
cited In Appendix A.
8.1.2. Oral. In a study conducted by Furedl et al. (1984a), 6- to
7-week-old F344 rats (75/sex/dose) were administered trinitrotoluene In the
diet at doses of 0, 0.4, 2, 10 or 50 mg/kg/day for 24 months. Administra-
tion of trinitrotoluene did not affect survival rate nor did It Induce signs
of toxldty throughout the study. Dose-related hematologlcal alterations
and hlstologlcal examination of organs and tissues Indicated that the MID
had been achieved. A significant Increase (p<0.01) In the combined
Incidence of urinary bladder papllloma and carcinoma (0/54, 0/54, 1/55 and
17/55 with Increasing doses of trinitrotoluene) was observed In female rats.
According to the Investigators, the historical Incidence of these tumors Is
low (<1%). The Incidences of hepatocellular (male rats), renal and urinary
bladder hyperplasla (female rats) were also Increased at the 50 mg/kg/day
dose level. Thus, the conclusion that trinitrotoluene 1s carcinogenic 1n
F344 rats under the experimental conditions used can be made.
In a study conducted by Furedl et al. (1984b), 4- to 5-week-old B6C3F1
hybrid mice (75/sex/dose) were administered trinitrotoluene 1n the diet at
doses of 0, 1.5, 10 or 70 mg/kg/day for 24 months. An MTD between 14 and
100 mg/kg/day had been previously estimated. A significant (p<0.05)
Increase )n the combined Incidence of leukemia/malignant lymphoma In the
spleen (9/45, 15/39, 17/37 and 21/33 with Increasing levels of trinitro-
toluene) was observed 1n females. However, the U.S. EPA (1988a) noted that
0240d -60- 04/23/90
-------
the Incidence of malignant lymphomas and lymphocytlc leukemia In all tissues
combined was not significantly Increased and concluded that the effect was
not chemical-related.
8.1.3. Other Routes. Pertinent data regarding the cardnogenlcHy of
trinitrotoluene administered by other routes were not located 1n the
available literature cited 1n Appendix A.
8.1.4. Weight of Evidence. No data were available regarding the cardno-
genldty of trinitrotoluene In humans. The animal cardnogenlcHy data are
limited to positive results In female F344 rats (Furedl et al., 1984a) and
negative results In B6C3F1 mice (Furedl et al., 1984b). Both studies were
well conducted with an adequate number of animals/sex/dose. Trinitrotoluene
was mutagenlc when tested In 5>. typhlmurlum. but was not genotoxlc 1n
mammalian cells j_n vivo. Based on the evidence discussed above, trinitro-
toluene has been assigned to U.S. EPA Group C: possible human carcinogen,
using the U.S. EPA (1986b) guidelines (U.S. EPA. 1988a. 1989).
8.1.5. Quantitative Risk Estimates.
8.1.5.1. INHALATION Pertinent data regarding cardnogenlcHy to
humans or animals of Inhalation exposure to trinitrotoluene were not located
In the available literature dted In Appendix A. U.S. EPA (1988a) did not
estimate a slope factor for Inhalation exposure or risk-specific levels 1n
air from the oral slope factor (Section 8.1.5.2.).
8.1.5.2. ORAL A human slope factor (q^) of 3.0xlO"2
(mg/kg/day)"1 was computed with the linearized multistage model by the
U.S. EPA (1988a, 1989) using bladder tumor Incidence data from female rats
In the study by Furedl et al. (1984a). The concentration of trinitrotoluene
1n drinking water associated with Increased lifetime risks of cancer are
0240d -61- 04/23/90
-------
0.01, 0.001 and 0.0001 mg/i, equivalent to 10, 1 and 0.1 iig/l, respec-
tively, at risk levels of 10~3, 10~* and 10~7. respectively. These
concentrations were calculated by dividing the given risk level by the q *
to obtain a risk specific dose and then multiplying the dose by the body
weight for humans (70 kg) and dividing by the reference dally water consump-
tion of 2 I for humans (U.S. EPA, 1980).
8.2. SYSTEMIC TOXICITY
8.2.1. Inhalation Exposure. Pertinent data regarding Inhalation exposure
to trinitrotoluene were not located In the available literature cited In
Appendix A.
8.2.2. Oral Exposure.
8.2.2.1. LESS THAN LIFETIME (SUBCHRONIC) ~ Several subchronlc
studies have been performed with trinitrotoluene using dogs, rats and mice.
In the study by Levlne et al. (1983), beagle dogs were treated with 0, 0.5,
2.0, 8.0 or 32 mg/kg/day In capsules for 26 weeks. Hepatocytlc cloudy
swelling and Increased liver weight were reported at 0.5 mg/kg/day (Rec. #1,
Appendix C). The severity of the lesions Increased with Increasing doses of
trinitrotoluene. Furthermore, higher doses of trinitrotoluene Induced
hematologlcal and clinical chemistry alterations and death. The lowest dose
of trinitrotoluene tested, 0.5 mg/kg/day, was Identified as a LOAEL.
In the study by Levlne et al. (1984a), F344 rats were administered doses
of 0, 1. 5, 125 or 300 mg/kg/day In the diet for 13 weeks. Rats at 25
mg/kg/day had a significant Increase 1n liver weight (Rec. #10, Appendix C).
Male rats, at this dietary level of trinltrololuene, showed a significant
decrease 1n the hematocrlt, hemoglobin concentration and erythrocyte counts.
These changes are Indicative of anemia. Effects seen at higher doses
Included hlstologlcal alterations In the testes and cerebellum (Rec. #11,
0240d -62- 04/23/90
-------
Appendix C). A dose of 5 mg/kg/day of trinitrotoluene was without adverse
effects and represent a NOAEL 1n this study (Rec. #9, Appendix C).
Dllley et al. (1982) conducted subchronlc studies using dogs, rats and
mice. Dogs were treated with trinitrotoluene In capsules at 0, 0.2, 2.0 or
20 mg/kg/day for <13 weeks. The lowest dose was a NOEL; red colored urine
and splenic effects were reported at 2.0 and 20 mg/kg/day, respectively.
Because of the small number of dogs used, the results cannot be used for
risk assessment. Rats were fed diets containing 0, 0.002, 0.01, 0.05 or
0.25% trinitrotoluene for <13 weeks. The high group showed depressed body
weight gain, Increased spleen weight and testlcular atrophy with hyperplasla
and anemia (Rec. #14, Appendix C). The 0.05% diet (equivalent to a dose of
34.7 mg/kg/day In the males and 36.4 mg/kg/day In the females) did not
appear to cause adverse effects and Is -designated as a NOAEL (Rec. #13,
Appendix C). Mice were fed diets containing 0, 0.001, 0.005, 0.025 or
0.125% trinitrotoluene for <13 weeks. High-group mice showed Increased
spleen and liver weights, occasional hepatocellular necrosis In some male
mice and mild hematologlcal changes (Rec. #15, Appendix C). No adverse
i
effects were reported at 0.05% In the diet, equivalent to 35.7 mg/kg/day 1n
the males and 37.8 mg/kg/day In the females (Rec. #16, Appendix C).
The three studies considered above appear comparable In quality.
Examination of the these three studies clearly Indicates that dogs are the
most sensitive species. The study of dogs (Levlne et al., 1983) has served
as the basis for deriving the verified chronic oral RfD (U.S. EPA, 1988b).
The LOAEL Identified 1n dogs, 0.5 mg/kg/day, is an order of magnitude lower
than the NOEL for F344 rats (Levlne et al., 1984a) and remains the most
appropriate basis for estimation of an RfD for subchronlc oral exposure.
0240d -63- 04/23/90
-------
In an earlier analysis of these data, U.S. EPA (1989) concluded that the
dog LOAEL of 0.5 mg/kg/day was the most appropriate basis from the sub-
chronic data for the longer-term HA. Application of an uncertainty factor
of 1000 (10 to estimate a NOAEL from a LOAEL, 10 to extrapolate from animals
to humans and 10 to provide additional protection for more sensitive members
of the population) would result In an RfO for subchronlc oral exposure of
0.0005 mg/kg/day. Because this value Is Identical to the RfD for chronic
oral exposure, U.S. EPA (1989) conservatively adopted the chronic RfO to
serve as the basis for the longer-term HA. To maintain consistency with the
U.S. EPA (1989) analysis, and because this current evaluation has revealed
no additional data or changes In methodology to suggest that another
approach would be more appropriate, the RfD of 0.0005 mg/kg/day .for chronic
oral exposure (Section 8.2.2.2.) 1s adopted as the RfO for subchronlc oral
exposure. Confidence in the key study, data base and RfO are medium as
discussed In Section 8.2.2.2.
8.2.2.2. CHRONIC Two studies that examined the chronic toxlclty of
trinitrotoluene were Identified. In the first study (Furedl et al., 1984a),
F344 rats were administered trinitrotoluene 1n the diet at doses of 0, 0.4,
2, 10 or 50 mg/kg/day for 24 months. A dose of 2 mg/kg/day was associated
with hypertrophy of the proximal convoluted tubules In the kidney and spleen
congestion (Rec. #5, Appendix C). In addition, females receiving the 2
mg/kg/day dose had sternal bone marrow flbrosls. Higher doses Induced
reduction 1n body weight gain, anemia, Increase In kidney and liver weights
and urinary bladder lesions in females. A dose of 0.4 mg/kg/day was
Identified as a NOEL In this study (Rec. #4, Appendix C).
Doses of 0, 1.5, 10 or 70 mg/kg/day were provided In the diet to B6C3F1
mice for 24 months (Furedl et al., 1984b). The dose of 70 mg/kg/day caused
0240d . -64- 04/23/90
-------
a significant decrease In body weight gain, mild transient anemia and
Increased liver weight (Rec. #8, Appendix C). A dose of 10 mg/kg/day was
without significant adverse effects (Rec. #7, Appendix C.).
The NOEL of 0.4 mg/kg/day from the Furedl et al. (1984a) rat study 1s
slightly lower than the LOAEL of 0.5 mg/kg/day from the subchronlc dog study
(Levlne et al., 1983). Other subchronlc studies using rats and dogs
demonstrate that dogs are the more sensitive species; therefore, It 1s
appropriate to base the RfD for chronic oral exposure on the LOAEL of 0.5
mg/kg/day In dogs, When considered with the NOEL of 0.4 mg/kg/day In rats,
U.S. EPA (1989) Concluded that the LOAEL of 0.5 mg/kg/day 1n dogs was near
'*.
the threshold for adverse effects and suggested that an uncertainty factor
of 1000 would be sufficient to provide for Inter- and Intraspedes extrapo-
lation, estimation of a NOAEL from a LOAEL and expansion from subchronlc to
chronic exposure. Applying the uncertainty factor of 1000 results 1n an RfD
for chronic oral exposure of 0.0005 mg/kg/day. This RfO has been verified
and 1s available on IRIS (U.S. EPA, 1988b). U.S. EPA (1988b) considered
confidence In the key study to be medium, with the only criticism that
administration of the test substance by capsule was not Ideal. Confidence
1n the data base 1s medium because there are no data on reproductive
effects. The subchronlc and chronic data support the magnitude of the RfO.
Confidence 1n the RfD 1s medium.
0240d -65- 04/23/90
-------
9. REPORTABLE QUANTITIES
9.1. BASED ON SYSTEMIC TOXICITY
The toxldty of 2,4,6-tMnltrotoluene was discussed In Chapter 6 and
dose-response data considered for CS derivation are summarized In Table 9-1.
Absent from Table 9-1 are the subchronlc dog studies by Dllley et al. (1982)
and Hart (1974) and the study using monkeys by Martin (1974) that used too
few animals for meaningful analysis. Hyperplasla of the epithelium of the
urinary bladder 1n female rats at 10 mg/kg/day 1n the 24-month dietary study
by Furedl et al. (1984a) 1s not Included because of the likelihood that this
was a preneoplastlc lesion. All studies 1n Table 9-1 examined similar
endpolnts In the species tested, which contributes to a better comparative
assessment of the overall toxldty of trinitrotoluene.
Effects attributed to subchronlc and chronic exposure to trinitrotoluene
are mortality (RV =10), lethargy and ataxla (RV =8), depressed body
weight gain or altered organ weights (RV =4), anemia (RV =5), kidney or
liver 'hypertrophy (RV =3) and splenic congestion, bone marrow Mbrosls or
cloudy swelling In liver hepatocytes (RV «5) (Furedl et. al., 1984a;
Levlne et al., 1983, 1984a). Based on estimated human equivalent doses
calculated for these effects, the liver alterations 1n dogs (Levlne et al.,
1983)'appear to be most sensitive endpolnt.
CSs and the corresponding RQs are calculated 1n Table 9-2 for the
effects Identified In Table 9-1. Data selection for Inclusion In Table 9-2
Include the lowest human equivalent dose associated with mortality, as well
as the lowest human equivalent dose associated with each of the less severe
effects. In the derivation of the CSs from subchonlc studies, an uncer-
tainty factor of 10 was applied to expand from subchronlc to chronic
exposure. From the studies presented in Table 9-2, the highest CS of 25.5,
0240d -66- 04/23/90
-------
TABU 9-1
Toxtctty Summary for 2.4.6-Trlnttrotoluene
1
-j
1
CD
**
\
CO
\
o
Route
Oral/
gavage
Oral/
gavage
Oral/
gavage
Oral
Oral
Oral
Oral
Oral
Oral
Oral
Species/
Strain
dog/beagle
dog/beagle
dog/beagle
rat/F344
rat/F344
rat/F344
rat/
Sprague-
Oawley
rat/
Swiss-
Webster
rats/F344
rats/F344
Average
Sex No. at Height
Start (kg)
N.f 6/sex 9.5b
N.F 6/sex 9.2°
F 6 8.3b
M.F 10/sex 0.35C
M.F 10/sex 0.3SC
M.F 10/sex 0.35C
M.F 5/sex 0.26b
M.F 5/sex 0.031b
M.F 75/sex 0.29b
M.F 75/sex 0.25b
Vehicle/
Physical
State
gelatin
capsules
gelatin
capsules
gelatin
capsules
food
food
food
food
food
food
food
Purity Exposure
99. IX 0.5 mg/kg/day
for 26 weeks
99. IX 8.0 mg/kg/day
for 26 weeks
99. IX 32.0 mg/kg/day
for 26 weeks
99. IX 25 mg/kg/day
for 13 weeks
99. IX 125 mg/kg/day
for 13 weeks
99. IX 300 mg/kg/day
for 13 weeks
>99.0X 0.2SX In diet
for 13 weeks
>99.0X 0.125X In diet
for 13 weeks
>99X 2.0 mg/kg/day
for 24 months
>99X 10 mg/kg/day
for 24 months
Transformed
Animal Dose
(rog/kg/day)
0.5
B.O
32.0
25. Od
125«
300d
162e
190«
2.0d
10. Od
equivalent
Human Dose*
(mg/kg/day)
0.26
4.1
15.7
4.3
21.4
51.3
25.1
14.5
0.32
1.5
Response
Anemia; hepatocyllc cloudy
swelling and Increased liver
weight
Liver cirrhosis, met hemoglobin-
em la spleen congestion and In-
crease In spleen weight
Death
Increased liver weight; hema to-
logical signs of anemia
Decreased body weight gain;
degeneration of testes; liver
hyperplasla; spleen congestion.
ataxla
Deaths from severe anemia
Decreased body weight gain;
Increase In organ weights;
testtcular atrophy
Liver necrosis, hepatomegaly.
splenomegaly
Kidney hypertrophy; spleen
congestion, bone narrow flbro-
sls In females
Decreased body weight gain;
methemogloblnemla; anemia;
Increased liver and kidney
weights; spleen congestion
Reference
Levlne et
al.. 1983
Levlne et
al.. 1983
Levlne et
al.. 1983
Levlne et
al., 1984a
Levlne et
al.. 1984a
Levtne et
al., 19B4a
Ollley et
al.. 1982
Ollley et
al.. 1982
Furedt et
al.. 1984a
Furedt et
al.. 1984*
-------
lAULf 9-1 (cont.)
0
IV)
-C*
CD
a.
Route Species/ Sex No. at
Strain Start
Oral ntce/B6C3M H.f JS/sex
hybrid
Average Vehicle/
Height Physical Purity Exposure
(kg) State
0.03° food >9B.BK JO mg/kg/day
for 24 months
Transformed
Animal Dose
(mg/kg/day)
jfjd
(qulvalenl
Human Dose'
(ag/kg/day)
5.3
Response
Decreased body weight gain;
lid anemia; changes In blood
chemistry. Increased liver
weight
Reference
Furedl et
al.. 19B4b
alnterspecles extrapolation Is performed by aultlplytng the animal dote expressed as ng/kg/day by a body surface area scaling factor.
^Calculated from data provided by the Investigators
Reference body weight from U.S. (PA (1980)
^Reference food consumption: rat (O.OS x body weight); nouse (0.11 x body weight) Iron U.S. (PA (19BO)
eEstlmated by the Investigators (average of values for males and females)
,
co
I
CO
-------
CD
I\J
O
Q.
TABLE 9-2
Composite Scores for Oral Toxlclty of 2,4,6-TMnUrotoluene
Chronic
Species Animal Dose Human HEOa RVd Effect RVe
(mg/kg/day) (mg/day)
Rat 2.0 22.4 3.5 Kidney hypertrophy; 5
spleen congestion;
bone marrow flbrosls
Dog 0.5 1.8° 5.1 HepatocyMc cloudy 5
swelling and In-
creased liver weight
i
* Dog 32.0 110 2.4 Death 10
Rat 10 105 2.5 Increased liver 5
weight; signs of
anemia; me t nemo -
globlnemla
Rat 125 150b 2.5 Ataxla 8
CS RQ Reference
17.5 1000 Furedl
et al..
25.5 100 Levlne
et al..
24 100 Levlne
et al..
12.5 1000 Furedl
et al..
17.6 1000 Levlne
et al..
1984a
1983
1983
1984a
1984a
aCalculated by multiplying the human equivalent dose by 70 kg to present the HEO In terms of mg/day for
a 70 kg human.
bThe dose was divided by an uncertainty factor of 10 to approximate chronic exposure.
u>
o
-------
which corresponds to an RQ of 100, 1s chosen to represent the hazard
associated with chronic (noncancer) toxlclty resulting from exposure to
trinitrotoluene (Table 9-3).
9.2. BASED ON CARCINOGENICITY
As discussed In Chapter 6, trinitrotoluene has been shown to cause
urinary bladder paplllomas and carcinomas In female rats (Furedl et al.,
1984a), but was not carcinogenic when tested 1n mice (Furedl et al., 1984b).
Trinitrotoluene was appropriately assigned to U.S. EPA Group C because of
limited evidence of cardnogenldty In animals and lack of human data. The
data for trinitrotoluene-Induced urinary bladder tumors In female F344 rats
(Furedl et al., 1984a) was used by the U.S. EPA (1988a, 1989) to calculate
an oral slope factor (q *) of 3.0xlO~2 (mg/kg/day)'1.' Using the data
on bladder tumor Incidence and the GLOBAL 82 version of the multistage model
(Howe and Crump, 1982), an F factor of 0.1856 (mg/kg/day)'1 was derived
(Table 9-4). Trinitrotoluene 1s therefore assigned to Potency Group 3,
which corresponds to a Hazard Ranking of LOW and a cancer-based RQ of 100.
0240d -70- 04/23/90
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TABLE 9-3
2,4,6-TMnltrotoluene
(CAS No. 118-96-7)
Minimum Effective Dose (MED) and Reportable Quantity (RQ)
Route: oral/gelatin capsules
Species/Sex: dogs/male and female
Dose*: . 1.8
Duration: 26 weeks
Effect: Increased liver weight and cloudy swelling 1n
hepatocytes
RVd: 5.1
RVe: 5.0
CS: 25.5
RQ: 100
Reference: Levlne et al., 1983
*Equ1valent human dose
0240d -71- 04/23/90
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TABLE 9-4
Derivation of Potency Factor (F) for 2,4,6-Tr1n1trotoluene
Reference:
Exposure route:
Species:
Strain:
Sex:
Vehicle or physical state:
Body weight:
Duration of treatment:
Duration of study:
Ufespan of animal:
Target organ:
Tumor type:
Experimental doses/exposures:
Transformed doses (mgAg/day)
Tumor Incidence:
Unadjusted 1/ED10:
Adjusted 1/ED-|0 (F Factor):
Fured! et al.f 1984a
oral/food
rat
F344
female
food
0.30 kg
24 months
24 months
24 months
urinary bladder
papllloma and carcinoma
0 0.4 2.0
0 0.4 2.0
0/54 0/54 0/55
NA
0.185632 (mg/kg/day)"1
10.0
10.0
1/55
50.0
50.0
17/55
NA = Not applicable
0240d
-72-
04/23/90
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APPENDIX A
LITERATURE SEARCHED
This HEED Is based on data Identified by computerized literature
searches of the following:
CHEMLINE
TSCATS
CASR online (U.S. EPA Chemical Activities Status Report)
TOXLINE
TOXLIT
TOXLIT 65
RTECS
OHM TADS
STORET
SRC Environmental Fate Data Bases
SANSS
AQUIRE
TSCAPP
NTIS
Federal Register
CAS ONLINE (Chemistry and Aquatic)
HSDB
SCISEARCH
Federal Research In Progress
These searches were conducted 1n April, 1989, and the following secondary
sources were reviewed:
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ACGIH (American Conference of Governmental Industrial Hyg1en1sts).
1987. TLVs: Threshold Limit Values for Chemical Substances In the
Work Environment adopted by ACGIH with Intended Changes for
1987-1988. Cincinnati, OH. 114 p.
Clayton, G.D. and F.E. Clayton, Ed. 1981. Patty's Industrial
Hygiene and Toxicology, 3rd rev. ed.. Vol. 2A. John Wiley and
Sons, NY. 2878 p.
Clayton, G.D. and F.E. Clayton, Ed. 1981. Patty's Industrial
Hygiene and Toxicology, 3rd rev. ed., Vol. 28. John Wiley and
Sons, NY. p. 2879-3816.
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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.
Grayson, M. and 0. Eckroth, Ed. 1978-1984. Klrk-Othmer Encyclo-
pedia of Chemical Technology, 3rd ed. John Wiley and Sons, NY. 23
Volumes.
Hamilton, A. and H.L. Hardy. 1974. Industrial Toxicology, 3rd ed.
Publishing Sciences Group, Inc., Littleton, MA. 575 p.
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graphs on the Evaluation of Carcinogenic Risk of Chemicals to
Humans. IARC, WHO, Lyons, France.
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EPA 600/6-84-010. NTIS PB84-243906. SRI International, Menlo
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Ouellette, R.P. and J.A. King. 1977. Chemical Week Pesticide
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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.
0240d -88- 04/23/90
-------
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, 0. 1971. Ecological Effects of Pesticides on Non-Target
Spedes. Prepared for the U.S. EPA, Washington, DC. PB-269605.
Schneider, B.A. 1979. Toxicology Handbook. Mammalian and Aquatic
Data. Book 1: Toxicology Data. Office of Pesticide Programs, U.S.
EPA, Washington, DC. EPA 540/9-79-003. NTIS PB 80-196876.
0240d -89- 04/23/90
-------
o
IM
O
Q.
APPENDIX B
Summary lable for 2,4.6-TMnl trotoluene
1
we
o
1
CD
A
Species
Inhalation Exposure
SubchronU ID
Chronic ID
Carctnogenlctty 10
Oral Exposure
Subchrontc dog
Chronic dog
Carctnogentctty rat
IMPORTABLE QUANTITIES
Based on Chronic Toxtctty: 100
Based on Carctnogentctty: 100
Exposure
10
ID
10
O.S mg/kg/day tn gelattn
capsules for 26 weeks
O.S ng/fcg/day tn gelatin
capsules for 26 weeks
0-50 rog/kg/day tn the
dtet for 24 months
Effect RfO or q\* Reference
10 10 NA
10 10 NA
10 10 NA
riepatocyttc cloudy swelling O.SxlO'* og/kg/ddy Levtne
and Increased Itwer weight et al..
hepatocyttc cloudy swelling O.SxlO** mg/kg/day Levlne
and Increased liver weight et al..
urinary bladder paptllomas 3.0xlO~* furedt
and carcinomas tn females (rag/kg/day)"1 et al..
Levtne
et al..
furedt
et al..
1983
1983
1984a
1983
1984a
CO
\
o
10 « Insufficient data; NA = not applicable
-------
APPENDIX C
DOSE/DURATION RESPONSE GRAPHS FOR EXPOSURE TO TRINITROTOLUENE
C.I. DISCUSSION
Dose/duration-response graphs for oral exposure to trinitrotoluene
generated by the method of Crockett et al. (1985) using the computer soft-
ware by Durkln and Meylan (1988) developed under contract to ECAO-C1ndnnat1
are presented 1n Figures C-l and C-2. Data used to generate these graphs
are presented In Section C.2. In the generation of these figures, all
responses are classified as adverse (FEL, AEL or LOAEL) or nonadverse (NOEL
or NOAEL) for plotting.
For oral exposure, the ordlnate expresses dosage as human equivalent
dose. Interspecles extrapolation Is performed by multiplying the animal
dosage In mg/kg/day by a scaling factor (the cube root of the ratio of the
animal:human body weight) to adjust for species differences In basal
metabolic rate (Mantel and Schnelderman, 1975). The result Is then
multiplied by 70 kg, the reference human body weight, to express the human
equivalent dose as mg/day for a 70 kg human.
The boundary for adverse effects (solid line) Is drawn by Identifying
the lowest adverse effect dose or concentration at the shortest duration of
exposure at which an adverse effect occurred. From this point, an Infinite
line 1s extended upward parallel to the dose axis. The starting point Is
then connected to the lowest adverse effect dose or concentration at the
next longer duration of exposure' that has an adverse effect dose or concen-
tration equal to or lower than the previous one. This process Is continued
to the lowest adverse effect dose or concentration. From this point a line
1s extended to the right parallel to the duration axis. The region of
adverse effects lies above the adverse effects boundary.
0240d -91- 04/23/90
-------
\
?
I
u
If.
0
c
u.
z
-------
<
t
\
u
0
a
z
c
Z
10000 f
1000 -t-
100 --
10 T
19
L10
LI 5
H9
n!6
L8
L5
^ ««.M4
» **^i
0.001
Exposure)
1
0.01 0.1
HUMAN EQUIU DURATION (frAction
CENSORED DATA METHOD
n
L
A
F
N
NOAEL
LOAEl
AEL
PEL
NOEL
FIGURE C-2
Dose/Duration - Response Graph for Oral Exposure to
Trinitrotoluene: Censored Data Method
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04/23/90
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Using the envelope method, the boundary for no adverse effects (dashed
line) 1s drawn by Identifying the highest no adverse effects dose or concen-
tration. From this point, a line parallel to the duration axis Is extended
to the dose or concentration axis. The starting point Is then connected to
the next lower or equal no adverse effect dose or concentration at a longer
duration of exposure. When this process can no longer be continued, a line
Is dropped parallel to the dose or concentration axis to the duration axis.
The no adverse effects region lies below the no adverse effects boundary.
At either end of the graph between the adverse effects and no adverse
effects boundaries are regions of ambiguity. -The area (If any) resulting
from Intersection of the adverse effects and no adverse effects boundaries
1s defined as the region of contradiction.
In the c-ensored data method, all no adverse effect points located 1n the
region of contradiction are dropped from consideration and the no adverse
effect boundary 1s redrawn so that It does not intersect the adverse effects
boundary and no region of contradiction 1s generated. This method results
1n the most conservative definition of the no adverse effects region.
Figures C-l and C-2 show the dose/duration-effects graphs generated by
the envelope and censored data methods for oral exposure to trinitrotoluene,
respectively. The boundary for adverse effects Is defined by three FELs
(F17, F18 and F19) corresponding to LD.Qs 1n mice and rats In the Ollley
et al. (1982) study and a LOAEL (LI) from a subchronlc study 1n dogs (Levlne
et al., 1983). N4, which Is below the line for adverse effects, and corre-
sponds to a NOEL in rats (Furedl et al., 1984a), was not used as the basis
for deriving the chronic oral RfD because dogs are clearly more sensitive
than rats. The area of contradiction results from the relative 1nsens1t1v-
1ty of rats and mice compared with dogs. The verified chronic RfO of
5xlO~4 mg/kg/day Is well below the boundary for adverse effects.
0240d -94- 04/23/90
-------
C.2. DATA USED TO GENERATE OOSE/OURATION-RESPONSE GRAPHS
C.2.1. Oral Exposure.
Chemical Name:
CAS Number:
Document Title:
Document Number:
Document Date:
Document Type:
2,4,6-TMnHrotoluene
118-96-7
Health and Environmental
Trinitrotoluene
Pending
Pending
HEED
Effects Document for 2,4,6-
RECORD #1: Species: Dogs
Sex: Both
Effect: LOAEL
Route: Capsul
Number Exposed:
Number Responses:
Type of Effect:
.Site of Effect:
Severity Effect:
12
NR
WGTIN
LIVER
4
Dose:
Duration
Duration
12
NR
HEMAT
BLOOD
5
Exposure:
Observation:
12
NR
DEGEN
LIVER
5
0.500
26.0 weeks
26.0 weeks
Comment: Doses given: 0, 0.5, 2.0, 8.0, 32.0 mg/kg/day; mild anemia,
hepatocelluar cloudy swelling and Increased liver weight.
Citation: Levlne et al., 1983
RECORD #2: Species: Dogs
Sex: Both
Effect: AEL
Route: Capsul
Number Exposed:
Number Responses:
Type of Effect:
Site of Effect:
Severity Effect:
12
NR
HEMAT
BLOOD
5
Dose:
Duration
Duration
12
1
DEGEN
LIVER
5
Exposure
Observat
12
NR
WGTIN
SPLEN
4
;
1on:
12
NR
DEGEN
SPLEN
5
8.000
26.0 weeks
26.0 weeks
Comment: See previous record; methemogloblnemla, 1 dog with liver cir-
rhosis, splenic congestion.
Citation: Levlne et al.. 1983
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-95-
04/23/90
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RECORD #3:
Species:
Sex:
Effect:
Route:
Dogs
Both
PEL
Capsul
Dose:
Duration Exposure:
Duration Observation:
32.000
26.0 weeks
26.0 weeks
Number Exposed: 12
Number Responses: 2
Type of Effect: DEATH
SHe of Effect: BODY
Severity Effect: 10
Comment:
Citation:
RECORD #4:
Comment:
Citation:
RECORD #5:
See record #1; 2 dogs died before 17 weeks of treatment.
Levlne et al. . 1983
Species: Rats
Sex: Both
Effect: NOEL
Route: Food
Number Exposed:
Number Responses:
Type of Effect:
SHe of Effect:
Severity Effect:
Doses given: 0, 0.4,
Furedl et al., 1984a
Species: Rats
Sex: Both
Effect: LOAEL
Route: Food
Number Exposed:
Number Responses:
Type of Effect:
SHe of Effect:
Severity Effect:
150
0
HYPRT
KIDNY
3
2, 10,
150
NR
HYPRT
KIDNY
3
Dose:
Duration
Duration
150
0
OEGEN
SPLEN
5
Exposure:
Observation:
75
0
HISTO
BONE
5
0.400
24.0 months
24.0 months
50 mg/kg/day.
Dose:
Duration
Duration
150
NR
DEGEN
SPLEN
5
Exposure:
Observation:
75
NR
HISTO
BONE
5
2.000
24.0 months
24.0 months
Comment: See previous record;
spleen In both sexes,
Citation: Furedl et. al., 1984a
kidney hypertorphy
bone marrow flbrosls
and congestion
1n females.
of
0240d
-96-
04/23/90
-------
RECORD #6:
Comment:
Citation:
Species:
Sex:
Effect:
Route:
Rats
Both
AEL
Food
Number Exposed:
Number Responses:
Type of Effect:
SHe of Effect:
Severity Effect:
See previous record;
Increased liver
methemog1ob1nem1a.
Dose:
Duration Exposure:
Duration Observation:
10.000
24.0 months
24.0 months
150
NR
HYPRT
KIDNY
3
150
NR
DEGEN
SPLEN
5
75
NR
HISTO
BONE
5
150
NR
WGTIN
LIVER
4
150
NR
HEMAT
BLOOD
5
additional effects at this
and kidney weight,
dose Include
anemia and
Furedl et. al.. 1984a
RECORD #7:
Comment:
Citation:
RECORD #8:
Comment:
Citation:
Species: Mice
Sex: Both
Effect: NOAEL
Route: Food
Number Exposed:
Number Responses:
Type of Effect:
Site of Effect:
Severity Effect:
150
0
HEMAT
BLOOD
5
Doses given: 0, 1.5, 10,
gain was significant only
Furedl et al., 1984b
Species: Mice
Sex: Both
Effect: LOAEL
Route: Food
Number Exposed:
Number Responses:
Typfc of Effect:
Site of Effect:
Severity Effect:
See previous record;
Furedl et al., 1984b
150
0
HEMAT
BLOOD
5
mild
Dose:
Duration
Duration
150
NR
WGTDC
BODY
4
Exposure:
Observation:
150
0
WGTIN
LIVER
4
70 mg/kg/day; decreased
at higher level .
Dose:
Duration
Duration
150
NR
WGTDC
BODY
4
anemia and
Exposure:
Observation:
150
0
WGTIN
LIVER
4
altered blood
10.000
24.0 months
24.0 months
body weight
70.000
24.0 months
24.0 months
chemistry.
0240d
-97-
04/23/90
-------
RECORD #9:
Comment:
Citation:
RECORD #10:
Species: Rats
Sex: Both
Effect: NOEL
Route: Food
Number Exposed:
Number Responses:
Type of Effect:
Site of Effect:
Severity Effect:
Doses of 0, 1, 5, 25
Levlne et al., 1984a
Species: Rats
Sex: Both
Effect: LOAEL
Route: Food
Number Exposed:
Number Responses:
Type of Effect:
Site of Effect:
Severity Effect:
20
0
WGTIN
LIVER
4
, 125,
20
NR
WGTIN
LIVER
4
Dose:
Duration
Duration
20
0
HEMAT
BLOOD
5
Exposure:
Observation:
20 10
0 0
FUND OEGEN
CNS TESTE
8 6
5.000
13.0 weeks
13.0 weeks
20
0
DEATH
BODY
10
300 mg/kg/day.
Dose:
Duration
Duration
10
NR
HEMAT
BLOOD
5
Exposure:
Observation:
20 10
0 0
FUND DEGEN
CNS TESTE
8 6
25.000
13.0 weeks
13.0 weeks
20
0
DEATH
BODY
10
Comment: See previous record; Increased liver weight In both sexes,
anemia only In males.
Citation: Levlne et al., 1984a
RECORD #11:
Comment:
Citation:
Species: Rats
Sex: Both
Effect: AEL
Route: Food
Number Exposed:
Number Responses:
Type of Effect:
Site of Effect:
Severity Effect:
See previous record;
Levlne et al., 1984a
20
NR
WGTIN
LIVER
4
anemia,
Dose:
Durat
Durat
20
NR
HEMAT
BLOOD
5
1on Exposure:
1on Observatl
20
NR
FUND
CNS
8
1
1
on: 1
10
NR
DEGEN
TESTE
6
25.000
3.0 weeks
3.0 weeks
20
0
DEATH
BODY
10
ata~x1a In both sexes.
0240d
-98-
04/23/90
-------
RECORD #12:
Comment:
Citation:
RECORD #13:
Species: Rats
Sex: Both
Effect: PEL
Route: Food
Number Exposed:
Number Responses:
Type of Effect:
Site of Effect:
Severity Effect:
See previous record;
Levlne et al.f 1984a
Species: Rats
Sex: Both
Effect: NOAEL
Route: Food
Number Exposed:
Number Responses:
Type of Effect:
Site of Effect:
Severity Effect:
20
NR
WGTIN
LIVER
4
deaths
10
0
HEMAT
BLOOD
5
Dose:
Duration
Duration
20
NR
HEMAT
BLOOD
5
attributed
Dose:
Duration
Duration
5
0
ATROP
TESTE
5
300
Exposure: 13.
Observation: 13.
20 10
NR NR
FUND DEGEN
CNS TESTE
8 6
to severe anemia
35.
Exposure: 13.
Observation: 13.
10 10
0 0
WGTDC WGTIN
BODY SPLEN
4 4
.000
0 weeks
0 weeks
20
2
DEATH
BODY
10
600
0 weeks
0 weeks
Comment: Dietary levels used: 0, 0.002, 0.01, 0.05, 0.25X
corresponding to doses (average male and female)of 0, 1.4,
7.2, 35.6, 162 mg/kg/ day; red urine during exposure.
Citation: Dllley et al., 1982
RECORD #14: Species:
Sex:
Effect:
Route:
Rats
Both
LOAEL
Food
Dose:
Duration Exposure:
Duration Observation:
162.000
13.0 weeks
13.0 weeks
Comment:
Citation;
10
NR
HEMAT
BLOOD
5
5
NR
ATROP
TESTE
5
10
NR
WGTDC
BODY
4
10
NR
WGTIN
SPLEN
4
Number Exposed:
Number Responses:
Type of Effect:
Site of Effect:
Severity Effect:
See previous record; 0.25% 1n diet; decreased
gain accompanied by decreased food consumption.
Dllley et al., 1982
body weight
0240d
-99-
04/23/90
-------
RECORD #15:
Comment:
Citation:
Species:
Sex:
Effect:
Route:
Mice
Both
LOAEL
Food
Dose:
Duration
Duration
10
0
WGTIN
LIVER
4
36
Exposure:
Observation:
10
0
WGTDC
BODY
4
10
5
PIGMN
SPLEN
5
1
1
3
3
.800
.0
.0
weeks
weeks
Number Exposed: 10
Number Responses: 0
Type of Effect: HEMAT
Site of Effect: BLOOD
Severity Effect: 5
Dietary levels of 0, 0.001, 0.005, 0.025, 0.125% correspond-
ing to doses (average for male and female) of 0, 1.6, 7.8,
36.8, 191 mg/kg/day; hemoslderosis of spleen.
Dllley et al.f 1982
RECORD #16:
Comment:
Citation:
RECORD #17:
Species: Mice
Sex: Both
Effect: NOAEL
Route: Food
Number Exposed: 10
Number Responses: 0
Type of Effect: HEMAT
Site of Effect: BLOOD
Severity Effect: 5
See previous record; red ur
Dllley et al., 1982
Species: Mice
Sex: Both
Effect: FEL
Route: Gavage
Dose:
Duration
Duration
10
0
WGTIN
LIVER
4
1ne.
Dose:
Duration
Duration
Exposure:
Observation:
10 10
0 0
WGTDC PIGMN
BODY SPLEN
4 5
Exposure:
Observation:
7.800
13.0 weeks
13.0 weeks
660.000
1 .0 days
14.0 days
Comment:
Citation:
Number Exposed: 20
Number Responses: NR
Type of Effect: DEATH
Site of Effect: BODY
Severity Effect: 10
Swiss-Webster mice were
administered in corn oil,
Ollley et al., 1982
used. The dose Is an LD5Q and was
0240d
-100-
04/23/90
-------
RECORD #18:
Comment:
Citation:
Species;
Sex:
Effect:
Route:
Rats
Female
PEL
Gavage
Dose:
Duration
Duration
Exposure:
Observation:
795.000
1.0 days
14.0 days
Number Exposed: 20
Number Responses: NR
Type of Effect: DEATH
SHe of Effect: BODY
Severity Effect: 10
Sprague-Dawley rats were used. The dose was administered In
corn oil and corresponds to an
Dllley et al., 1982
RECORD #19:
Species:
Sex:
Effect:
Route:
Rats
Hale
PEL
Gavage
Dose:
Duration
Duration
Exposure:
Observation:
1320.000
1.0 days
14.0 days
Comment:
Citation:
Number Exposed: 20
Number Responses:- NR
Type of Effect: DEATH
SHe of Effect: BODY
Severity Effect: 10
Sprague-Dawley rats were
administered 1n corn oil.
Dllley et al., 1982
used. The dose Is an 1059 and was
0240d
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04/23/90
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