1978 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

NO. 81 

BIOASSAY OF 

TRIMETHYLPHOSPHATE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 512-56-1 

NCI-CG-TR-81 

U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE 

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

TRIMETHYLPHOSPHATE
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

Bethesda, Maryland 20014
 

U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE
 

Public Health Service
 

National Institutes of Health
 

DHEW Publication No. (NIH) 78-1331
 

BIOASSAY OF
 

TRIMETHYLPHOSPHATE
 

For Possible Carcinogenicity
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institute of Health
 

FOREWORD: This report presents the results of the bioassay of
 

trimethylphosphate conducted for the Carcinogenesis Testing
 

Program, Division of Cancer Cause and Prevention, National Cancer
 

Institute (NCI), National Institutes of Health, Bethesda,
 

Maryland. This is one of a series of experiments designed to
 

determine whether selected environmental chemicals have the
 

capacity to produce cancer in animals. Negative results, in
 

which the test animals do not have a greater incidence of cancer
 

than control animals, do not necessarily mean that the test
 

chemical is not a carcinogen, inasmuch as the experiments are
 

conducted under a limited set of circumstances. Positive results
 

demonstrate that the test chemical is carcinogenic for animals
 

under the conditions of the test and indicate that exposure to
 

the chemical is a potential risk to man. The actual
 

determination of the risk to man from animal carcinogens requires
 

a wider analysis.
 

CONTRIBUTORS; The bioassay of trimethylphosphate was conducted
 

by Litton Bionetics, Inc., Kensington, Maryland, initially under
 

direct contract to NCI and currently under a subcontract to
 

Tracor Jitco, Inc., the prime contractor for the NCI
 

Carcinogenesis Testing Program.
 

The experimental design and doses were chosen by Drs. E. K.
 

Weisburger1, J. H. Weisburger1'2, N. P. Page1'3, and F. M.
 

Garner^. Administration of test chemical and observation of
 

animals were supervised by Dr. F. M. Garner, with the technical
 

assistance of Mr. R. Cypher^, Mr. H. D. Thornett^, and Mr. D. J.
 

Howard^.
 

iii
 
Histopathologic examinations of rats were performed by Drs. R. A.
 

Montali4, P. Hildebrandt4, N. J. Wosu4, and H. R. Seibold4 and
 

those of mice by Drs. Montali and Wosu. Histologic sections of
 

all tumors and hyperplasias were reexamined by Dr. Montali, who
 

also examined all diagnoses and prepared the interpretive
 

pathology summary included in this report.
 

Animal pathology tables and survival tables were complied at EG&G
 

Mason Research Institute-*. The statistical analyses were
 

performed by Dr. J. R. Joiner^ and Ms. P. L. Yong^, using methods
 

selected for the bioassay program by Dr. J. J. Gart'. Chemicals
 

used in this bioassay were analyzed under the direction of Dr. E.
 

Murrill , and the analytical results were reviewed by Dr. S. S.
 

Olin". The structural formula was supplied by NCI.
 

This report was prepared at Tracer Jitco" under the direction of
 

NCI. Those responsible for the report at Tracor Jitco were Dr.
 

Marshall Steinberg, Director of the Bioassay Program; Dr. L. A.
 

Campbell, Deputy Director for Science; Dr. J. F. Robens,
 

toxicologist; Dr. R. L. Schueler, pathologist; Dr. G. L. Miller
 

and Mr. W. D. Reichardt, bioscience writers; and Dr. E. W.
 

Gunberg, technical editor, assisted by Ms. Y. E. Presley and Ms.
 

P. J. Graboske.
 

The statistical analysis was reviewed by members of the
 

Mathematical Statistics and Applied Mathematics Section of NCI:
 

Dr. John J. Gart, Mr. Jun—mo Nam, Dr. Hugh M. Pettigrew, and Dr.
 

Robert E. Tarone.
 

The following other scientists at NCI were responsible for
 

evaluating the bioassay, interpreting the results, and reporting
 

the findings: Dr. Kenneth C. Chu, Dr. Cirpiano Cueto, Jr., Dr. J.
 

Fielding Douglas, Dr. Dawn G. Goodman, Dr. Richard A. Griesemer,
 

Dr. Harry A. Milman, Dr. Thomas W. Orme, Dr. Robert A. Squire^,
 

and Dr. Jerrold M. Ward.
 

iv
 
^Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

with the Naylor Dana Institute for Disease Prevention,
 

American Health Foundation, Hammond House Road, Valhalla,
 

New York.
 

with the National Institute for Occupational Safety and
 

Health, Rockville, Maryland.
 

^Litton Bionetics, Inc., 5516 Nicholson Lane, Kensington,
 

Maryland.
 

Mason Research Institute, 1530 East Jefferson Street,
 

Rockville, Maryland.
 

^Tracer Jitco, Inc., 1776 East Jefferson Street, Rockville,
 

Maryland.
 

'Mathematical Statistics and Applied Mathematics Section,
 

Biometry Branch, Field Studies and Statistics, Division of
 

Cancer Cause and Prevention, National Cancer Institute, National
 

Institutes of Health, Bethesda, Maryland.
 

"Midwest Research Institute, 425 Volker Boulevard, Kansas City,
 

Missouri.
 

"NOW with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

SUMMARY
 

A bioassay of trimethylphosphate for possible carcinogenicity was
 

conducted by administering the compound by gavage to Fischer 344
 

rats and B6C3F1 mice.
 

Groups of 50 rats and 50 mice of each sex were administered
 

trimethylphosphate in distilled water three times per week at one
 

of two doses, either 50 or 100 mg/kg body weight for the rats and
 

either 250 or 500 mg/kg body weight for the mice. Vehicle
 

controls consisted of groups of 20 rats and 20 mice of each sex.
 

The rats were dosed for 104 weeks and the mice for 103 weeks.
 

All surviving rats were killed at week 105 and all surviving mice
 

at week 103.
 

Mean body weights of dosed male and female rats and female mice
 

were slightly lower than those of the corresponding vehicle
 

controls throughout the study; mean body weights of the male mice
 

were comparable to those of the vehicle controls. Survival rates
 

of both rats and mice were high, and adequate numbers of animals
 

were at risk for the development of late-appearing tumors.
 

In the male rats, the incidence of fibromas of the subcutanous
 

tissue was higher (P = 0.036) in the high-dose group than in the
 

vehicle controls (control 0/20, low-dose 2/50, high-dose 9/49),
 

and there was a dose-related trend (P = 0.006) in the incidences
 

of these fibromas. In the female rats, no tumors occurred in the
 

dosed groups at significantly increased incidences, compared with
 

corresponding controls.
 

In the male mice, no tumors occurred in the dosed groups at
 

significantly increased incidences, compared with controls. In
 

the female mice, the incidence of adenocarcinomas of the
 

endometrium was higher (P = 0.004) in the high-dose group than in
 

the vehicle controls (controls 0/16, low-dose 7/40, high-dose
 

13/37), and there was a significant dose-related trend (P =
 

0.003) in the incidences of these adenocarcinomas.
 

vii
 
It is concluded that under the conditions of this bioassay,
 

trimethylphosphate was carcinogenic in female B6C3F1 mice,
 

inducing adenocarcinomas of the uterus/endometrium.
 

Trimethylphosphate was associated with the induction of benign
 

fibromas of the subcutaneous tissue in male Fischer 344 rats. No 

evidence of carcinogenicity of the compound was obtained in 

female rats or in male mice. 

viii
 
TABLE OF CONTENTS
 

I. Introduction 1
 

II. Materials and Methods 3
 

A. Chemical 3
 

B. Dosage Preparation 4
 

C. Animals 4
 

D. Animal Maintenance 5
 

E. Subchronic Studies 6
 

F. Designs of Chronic Studies 8
 

G. Clinical and Pathologic Examinations 8
 

H. Data Recording and Statistical Analyses 11
 

III. Results - Rats 17
 

A. Body Weights and Clinical Signs (Rats) 17
 

B. Survival (Rats) 17
 

C. Pathology (Rats) 20
 

D. Statistical Analyses of Results (Rats) 22
 

IV. Results - Mice 23
 

A. Body Weights and Clinical Signs (Mice) 23
 

B. Survival (Mice) 23
 

C. Pathology (Mice) 26
 

D. Statistical Analyses of Results (Mice) 29
 

V. Discussion » 31
 

VI. Bibliography 33
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms
 

in Rats Administered Trimethylphosphate
 

by Gavage 35
 

Table Al Summary of the Incidence of Neoplasms
 

in Male Rats Administered Trimethylphosphate
 

by Gavage 37
 

ix
 
Table A2 Summary of the Incidence of Neoplasms 

in Female Rats Administered Trimethylphosphate by Gavage 41 

Appendix B Summary of the Incidence of Neoplasms 

in Mice Administered Trimethylphosphate 

by Gavage 45 

Table Bl Summary of the Incidence of Neoplasms 

in Male Mice Administered Trimethylphosphate by Gavage 47 

Table B2 Summary of the Incidence of Neoplasms 

in Female Mice Administered 

Trimethylphosphate by Gavage 51 

Appendix C Summary of the Incidence of Nonneoplastic 

Lesions in Rats Administered 

Trimethylphosphate by Gavage 57 

Table Cl Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Administered 

Trimethylphosphate by Gavage 59 

Table C2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Administered 

Trimethylphosphate by Gavage.. 64 

Appendix D Summary of the Incidence of Nonneoplastic 

Lesions in Mice Administered 

Trimethylphosphate by Gavage 69 

Table Dl Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Administered 

Trimethylphosphate by Gavage 71 

Table D2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Administered 

Trimethylphosphate by Gavage 75 

Appendix E Analyses of the Incidence of Primary 

Tumors in Rats Administered 

Trimethylphosphate by Gavage 79 

x
 
Table El
 

Table E2
 

Appendix F
 

Table Fl
 

Table F2
 

Table 1
 

Table 2
 

Figure 1
 

Figure 2
 

Figure 3
 

Figure 4
 

Analyses of the Incidence of Primary 

Tumors in Male Rats Administered 

Trimethylphosphate by Gavage 81 

Analyses of the Incidence of Primary 

Tumors in Female Rats Administered 

Trimethylphosphate by Gavage 85 

Analyses of the Incidence of Primary 

Tumors in Mice Administered 

Trimethylphosphate by Gavage 89 

Analyses of the Incidence of Primary 

Tumors in Male Mice Administered 

Trimethylphosphate by Gavage 91 

Analyses of the Incidence of Primary 

Tumors in Female Mice Administered 

Trimethylphosphate by Gavage 94 

TABLES 

Design of Chronic Studies of 

Trimethylphospate in Rats 9 

Design of Chronic Studies of 

Trimethylphosphate in Mice 10 

FIGURES 

Growth Curves for Rats Administered 

Trimethylphosphate by Gavage 18 

Survival Curves for Rats Administered 

Trimethylphosphate by Gavage 19 

Growth Curves for Mice Administered 

Trimethylphosphate by Gavage 24 

Survival Curves for Mice Administered 

Trimethylphosphate by Gavage 25 

xi
 

I. INTRODUCTION
 

Trimethylphosphate (CAS 512-56-1; NCI C03781) is an alkylating
 

agent which has been used as a gasoline additive (Hinkamp and
 

Warren, 1958), a methylating agent (Billman et al., 1942), an
 

intermediate for the production of polymethyl polyphosphates
 

(Toy, 1949), and a flame retardant in polymers (Matsunaga et al. ,
 

1975). Although it is still available commercially, it is no
 

longer manufactured in the U.S. (Chem Sources - U.S.A., 1977).
 

Trimethylphosphate is known to cause sterility in mice, rats, and
 

rabbits (Harbison et al., 1976). It produces chromosomal abnormalities in the bone-marrow cells of rats (Adler et al. , 1971)
 

and dominant lethal effects in mice (Epstein, 1970).
 

Trimethylphosphate was selected for testing because its
 

production and industrial applications could result in the
 

exposure of workers in the chemical industry to the compound.
 

II. MATERIALS AND METHODS
 

A. Chemical
 

TRIMETHYLPHOSPHATE
 

CH3O — p — QCH3
 

I
 

OCH3
 

Trimethylphosphate was obtained in two batches (Lot Nos. 062927
 

and 112737) from Aldrich Chemical Company, Milwaukee, Wisconsin.
 

The identity and purity of both batches were checked by analysis.
 

Elemental analyses (C, H, P) were consistent with C^E^PO^, the
 

molecular formula of trimethylphosphate. Infrared spectra were
 

identical to those given in the literature (Aldrich Library of
 

Infrared Spectra, 1976). Thin-layer chromatography using silica
 

gel F-254(B) showed one major spot (with a slight trace at the
 

origin in Lot No. 062927). Lot No. 112737 was estimated to be
 

> 99% pure by vapor-phase chromatography (vpc). Lot No. 062927
 

contained approximately 2.4% methanol by vpc and nuclear magnetic
 

resonance, and 1.8% water by Karl Fisher analysis.
 
The chemical was stored at 4°C in the original containers.
 

B. Dosage Preparation
 

Test solutions were prepared with distilled water on each day
 

that a dose was to be administered. Excess solutions were not
 

stored.
 

At 25°C, the half-life for the hydrolysis of the first methyl
 

group in trimethylphosphate in water at neutral or slightly acid
 

pH is calculated to be approximately 53 days (by extrapolation
 

from Barnard et al., 1961). About 0.1% hydrolysis is expected in
 

24 hours. This is in essential agreement with the report of
 

Newell et al. (1977) and Spanggord (1977) that no hydrolysis of
 

aqueous solutions of trimethylphosphate was detected after 7 days
 

at ambient temperature. Thus, no significant hydrolysis of
 

trimethylphosphate in the test solutions for this bioassay should
 

have occurred.
 

C. Animals
 

Fischer 344 rats and B6C3F1 mice were obtained from Charles River
 

Breeding Laboratories, Inc., Wilmington, Massachusetts, under a
 

contract with the Division of Cancer Treatment, NCI. On arrival
 

at the laboratory, the animals were approximately 4 weeks of age.
 
All animals were quarantined for 2 weeks and were then assigned
 

to control or dosed groups.
 

D. Animal Maintenance
 

All animals were housed in temperature- and humidity-controlled
 

rooms. The temperature was maintained at 21-25°C and the
 

relative humidity at 45-55%. There were 15 changes of room ail
 

per hour, and the incoming and exhaust air was passed through
 

HEPA filters. The animal rooms were positively pressurized with
 

respect to an exit hall and negatively pressurized with respect
 

to an entrance hall. Cool white fluorescent lighting was
 

provided 8 hours per day. Ground Wayne® Lab Blox animal meal
 

(Allied Mills, Inc., Chicago, 111.) was available ad libitum and
 

was replenished three times per week. Tap water acidified with
 

HC1 to pH 2.5 also was available ad libitum.
 

Rats were housed four per cage and mice five per cage in solid
 

polycarbonate cages. Each cage was covered with a wire mesh
 

screen and a sheet of filter paper. Heat-treated hardwood chip
 

bedding (Absorb-Dri®, Lab Products, Garfield, N. J.) was used in
 

the cages. Cages and water bottles were sanitized twice per week
 

and feed hoppers once per week at approximately 82°C; bedding was
 

replaced twice per week.
 

Rats and mice were housed in separate rooms. Control animals
 
were housed in the same room as the respective dosed animals.
 

Animals administered trimethylphosphate were housed in the same
 

room as animals of the same species administered the following
 

chemicals:
 

RATS
 

Feed Studies
 

l-H-l,2,4-triazole-3-amine (amitrole) (CAS 61-82-5)
 

2,4-dimethoxybenzenamine hydrochloride (NCI C02255)
 

N'-(chloroacetyl)-N-phenylacetamide (CAS 140-49-8)
 

N-ethyl-N'-(5-nitro-2-thiazolyl)urea (nithiazide) (CAS 139-94-6)
 

MICE
 

Feed Studies
 

2,4-dimethoxybenzenamine hydrochloride (NCI C02255)
 

N'-(chloroacetyl)-N-phenylacetamide (CAS 149-99-8)
 

N-ethyl-N'-(5-nitro-2-thiazolyl)urea (nithiazide) (CAS 139-94-6)
 

1,4-benzenediamine dihydrochloride (CAS 624-18-0)
 

4-nitro-o-phenylenediamine (CAS 99-56-9)
 

2,4-dihydro-5-methyl-2-phenyl-3H-pyrazol-3-one (CAS 89-25-8)
 

Gavage Studies
 

3-(chloromethyl)pyridine hydrochloride (NCI C03838)
 

2-(chloromethyl)pyridine hydrochloride (CAS 4377-33-7)
 

3,3-dimethyl-2-oxethanone (pivalolactone) (CAS 1955-45-9)
 

E. Subchronic Studies
 

Subchronic studies were conducted to estimate the maximum
 

tolerated concentrations (hereinafter referred to as "low doses"
 

and "high doses") of trimethylphosphate, on the basis of which
 

low and high doses were determined for use in the chronic study.
 
Trimethylphosphate was administered by gavage in a vehicle of
 

distilled water three times per week for 7 weeks. Doses for rats
 

were 100, 147, 215, 316, 464, 681, 1,000, or 1,470 mg/kg body
 

weight. Doses for mice were 147, 215, 316, 464, 681, 1,000,
 

1,470, or 2,150 mg/kg. Five animals were tested at each dose.
 

Control groups consisted of five animals of each species and sex
 

and were administered distilled water by gavage on the same
 

schedule as dosed groups. All surviving animals were killed and
 

necropsied 1 week after the end of the period of administration
 

of the trimethylphosphate.
 

In the rats, all males and females died when administered doses
 

of 681 mg/kg and greater, and one male died at a dose of 464
 

mg/kg. At the end of the period of administration, the gains in
 

mean body weight in males and females at doses up to and
 

including 316 mg/kg were approximately 80% of those of the
 

controls. At 464 mg/kg, the gain in mean weight in males was 56%
 

of that in controls; in females, it was 68% of that in controls.
 

Rats dying before the end of the study had distended bladders and
 

gastrointestinal hemorrhages. Low and high doses for the chronic
 

studies using rats were set at 50 and 100 mg/kg.
 

In the mice, five males and one female died at a dose of 2,150
 

mg/kg, and two females died at 1,470 mg/kg. There were no other
 

deaths. Gains in mean body weight were slightly depressed at
 
doses of 681 rag/kg and above in males, but were not greatly
 

affected in any of the groups of females. Low and high doses for
 

the chronic studies using mice were set at 250 and 500 rag/kg.
 

F. Designs of Chronic Studies
 

The designs of the chronic studies are shown in tables 1 and 2.
 

G. Clinical and Pathologic Examinations
 

All animals were observed twice daily for signs of toxicity, and
 

animals that were moribund were killed and necropsied. Rats and
 

mice were weighed individually at regular intervals. Palpation
 

for masses was carried out at each weighing.
 

The pathologic evaluation consisted of gross and microscopic
 

examination of all major tissues, organs, or gross lesions taken
 

from killed animals and from animals found dead. The following
 

tissues and organs were routinely subjected to microscopic
 

examination: skin, lymph nodes, mammary gland, salivary gland,
 

bone marrow, trachea, lungs and bronchi, heart, thyroid,
 

parathyroid, esophagus, stomach, small intestine, large
 

intestine, liver, gallbladder (mice), pancreas, spleen, kidney,
 

adrenal, urinary bladder, prostate or uterus, testis or ovary,
 

brain, and pituitary. Peripheral blood smears were prepared from
 

each animal whenever possible. Occasionally, additional tissues
 
Table 1. Design of Chronic Studies of Trimethylphosphate in Rats
 

Trimethyl-


Sex and Initial phosphate Time on Study
 

Test No. of Dose Dosed Observed
 

Group Animals3 (mg/kR)b (weeks) (weeks)
 

Males
 

Vehicle-Control0 20 0 105
 

Low-Dose 50 50 104 1
 

High -Dose 50d 100 104 1
 

Females
 

Vehicle-Control0 20 0 105
 

Low-Dose 50 50 104 1
 

High-Dose 50d 100 104 1
 

a

All animals were about 42 days of age when placed on study.
 

^Trimethylphosphate in distilled water was administered by gavage
 

three times per week. Doses were based on the mean body weight
 

of the group and were adjusted once per month.
 

°Vehicle controls received distilled water by gavage three times
 

per week.
 

"One animal in the high-dose male group was found to be a female,
 

and one in the high-dose female group was found to be a male.
 
Table 2. Design of Chronic Studies of Trimethylphosphate in Mice
 

Sex and 

Test 

G r oup 

Initial 

No. of 

Animals3 

Trimethylphosphate 

Dose 

(mg/kg)b 

Time on Study 

Dosed

(weeks)

 Observed 

 (weeks) 

Males 

Vehicle-Control0 20 0 103 

Low-Dose 50 250 103 0 

High-Dose 49 500 103 0 

Females 

Vehicle-Control0 20 0 103 

Low-Dose 50 250 103 0 

High -Dose 50d 500 103 0 

a

All animals were about 42 days of age when placed on study.
 

^Trimethylphosphate in distilled water was administered by gavage
 

three times per week. Doses were based on the mean body weight
 

of the group and were adjusted once per month.
 

G

Vehicle controls received distilled water by gavage three times
 

per week.
 

"One animal in the high-dose female group was found to be a male.
 

10
 
were also examined microscopically. The different tissues were
 

preserved in 10% buffered formalin, embedded in paraffin,
 

sectioned, and stained with hematoxylin and eosin. An occasional
 

section was subjected to special staining techniques for more
 

definitive diagnosis.
 

A few tissues from some animals were not examined, particularly
 

from those animals that died early. Also, some animals were
 

missing, cannibalized, or judged to be in such an advanced state
 

of autolysis as to preclude histopathologic evaluation. Thus,
 

the number of animals from which particular organs, tissues, or
 

lesions were examined microscopically varies and does not
 

necessarily represent the number of animals that were placed on
 

study in each group.
 

H. Data Recording and Statistical Analyses
 

Pertinent data on this experiment have been recorded in an automatic data processing system, the Carcinogenesis Bioassay Data
 

System (Linhart et al., 1974). The data elements include descriptive information on the chemicals, animals, experimental design,
 

clinical observations, survival, body weight, and individual
 

pathologic results, as recommended by the International Union
 

Against Cancer (Berenblum, 1969). Data tables were generated for
 

verification of data transcription and for statistical review.
 

11
 
These data were analyzed using the statistical techniques
 

described in this section. Those analyses of the experimental
 

results that bear on the possibility of carcinogenicity are
 

discussed in the statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

procedure of Kaplan and Meier (1958) and are presented in this
 

report in the form of graphs. Animals were statistically
 

censored as of the time that they died of other than natural
 

causes or were found to be missing; animals dying from natural
 

causes were not statistically censored. Statistical analyses for
 

a possible dose-related effect on survival used the method of Cox
 

(1972) for testing two groups for equality and Tarone's (1975)
 

extensions of Cox's methods for testing for a dose-related trend.
 

One-tailed P values have been reported for all tests except the
 

departure from linearity test, which is only reported when its
 

two-tailed P value is less than 0.05.
 

The incidence of neoplastic or nonneoplastic lesions has been
 

given as the ratio of the number of animals bearing such lesions
 

at a specific anatomic site (numerator) to the number of animals
 

in which that site is examined (denominator). In most instances,
 

the denominators included only those animals for which that site
 

was examined histologically. However, when macroscopic examination was required to detect lesions prior to histologic sampling
 

12
 
(e.g., skin or mammary tumors), or when lesions could have
 

appeared at multiple sites (e.g., lymphomas), the denominators
 

consist of the numbers of animals necropsied.
 

The purpose of the statistical analyses of tumor incidence is to
 

determine whether animals receiving the test chemical developed a
 

significantly higher proportion of tumors than did the control
 

animals. As a part of these analyses, the one-tailed Fisher
 

exact test (Cox, 1970) was used to compare the tumor incidence of
 

a control group with that of a group of dosed animals at each
 

dose level. When results for a number of dosed groups (k) are
 

compared simultaneously with those for a control group, a
 

correction to ensure an overall significance level of 0.05 may be
 

made. The Bonferroni inequality (Miller, 1966) requires that the
 

P value for any comparison be less than or equal to 0.05/k. In
 

cases where this correction was used, it is discussed in the
 

narrative section. It is not, however, presented in the tables,
 

where the Fisher exact P values are shown.
 

The Cochran-Armitage test for linear trend in proportions, with
 

continuity correction (Armitage, 1971), was also used. Under the
 

assumption of a linear trend, this test determines if the slope
 

of the dose-response curve is different from zero at the one-


tailed 0.05 level of significance. Unless otherwise noted, the
 

direction of the significant trend is a positive dose relation13
 
ship. This method also provides a two-tailed test of departure
 

from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation
 

of tumors. In this analysis, deaths that occurred before the
 

first tumor was observed were excluded by basing the statistical
 

tests on animals that survived at least 52 weeks, unless a tumor
 

was found at the anatomic site of interest before week 52. When
 

such an early tumor was found, comparisons were based exclusively
 

on animals that survived at least as long as the animal in which
 

the first tumor was found. Once this reduced set of data was
 

obtained, the standard procedures for analyses of the incidence
 

of tumors (Fisher exact tests, Cochran-Armitage tests, etc.) were
 

followed.
 

When appropriate, life-table methods were used to analyze the
 

incidence of tumors. Curves of the proportions surviving without
 

an observed tumor were computed as in Saffiotti et al. (1972).
 

The week during which an animal died naturally or was sacrificed
 

was entered as the time point of tumor observation. Cox's
 

methods of comparing these curves were used for two groups;
 

Tarone's extension to testing for linear trend was used for three
 

groups. The statistical tests for the incidence of tumors which
 

used life-table methods were one-tailed and, unless otherwise
 

14
 
noted, in the direction of a positive dose relationship.
 

Significant departures from linearity (P < 0.05, two-tailed test)
 

were also noted.
 

The approximate 95 percent confidence interval for the relative
 

risk of each dosed group compared with its control was calculated
 

from the exact interval on the odds ratio (Gart, 1971). The
 

relative risk is defined as Pt/pc where pt is the true binomial
 

probability of the incidence of a specific type of tumor in a
 

dosed group of animals and pc is the true probability of the
 

spontaneous incidence of the same type of tumor in a control
 

group. The hypothesis of equality between the true proportion of
 

a specific tumor in a dosed group and the proportion in a control
 

group corresponds to a relative risk of unity. Values in excess
 

of unity represent the condition of a larger proportion in the
 

dosed group than in the control.
 

The lower and upper limits of the confidence interval of the
 

relative risk have been included in the tables of statistical
 

analyses. The interpretation of the limits is that in approximately 95% of a large number of identical experiments, the true
 

ratio of the risk in a dosed group of animals to that in a
 

control group would be within the interval calculated from the
 

experiment. When the lower limit of the confidence interval is
 

greater than one, it can be inferred that a statistically
 

15
 
significant result (P < 0.025 one-tailed test when the control
 

incidence is not zero, P < 0.050 when the control incidence is
 

zero) has occurred. When the lower limit is less than unity, but
 

the upper limit is greater than unity, the lower limit indicates
 

the absence of a significant result while the upper limit
 

indicates that there is a theoretical possibility of the
 

induction of tumors by the test chemical, which could not be
 

detected under the conditions of this test.
 

16
 
III. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

Mean body weights of the low- and high-dose rats of each sex were
 

slightly lower than those of the corresponding vehicle controls,
 

and the depressions in weight were dose related (figure 1).
 

Fluctuation in the growth curve may be due to mortality; as the
 

size of a group diminishes, the mean body weight may be subject
 

to variation. No other chemical-related clinical signs were
 

reported.
 

B. Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female rats administered trimethylphosphate
 

by gavage at the doses of this bioassay, together with those of
 

the vehicle controls, are shown in Eigure 2.
 

The results of the Tarone test for positive dose-related trend in
 

mortality are not significant in either sex. All of the rats
 

lived beyond week 52 on study, and adequate numbers of animals of
 

each sex were at risk for development of late-appearing tumors.
 

In male rats, 17/49 (35%) of the high-dose group, 28/50 (56%) oi
 

the low-dose group, and 8/20 (40%) of the matched controls lived
 

to the end of the study. In females, the percentages of survival
 

17
 
D 

2 O D D ° DUD 

A O 0 O A A 8 A A A O ­

A A AA 

8

D 

s 

300 H 

LU 

5 

>- 250­ Q 

O 

m 

MALE RATS 

D VEHICLE CONTROL 

O LOW DOSE 

A HIGH DOSE 

40 50 GO 70 100 110 

TIME ON STUDY (WEEKS) 

i

 3°° D D D D 8 000 

2 & ^ A A 8 

A Q 

O 

m 

FEMALE RATS 

D VEHICLE CONTROL 

O LOW DOSE 

A HIGH DOSE 

—i 1 1 1— 40 SO 60 70 

TIME ON STUDY (WEEKS) 

Figure 1. Growth Curves for Rats Administered Trimethylphosphate by Gavage 

18 
PROBABILIT

Y OF SURVIVAL PROBABILIT

Y OF SUR

o o o o 

A CJ1 O> 

S o o o 
to termination of the study are 27/49 (55%) in high-dose group,
 

36/50 (72%) in low-dose group, and 12/20 (60%) in the matched
 

controls.
 

C. Pathology (Rats)
 

Histopathologic findings on neoplasms in rats are summarized in
 

Appendix A, tables Al and A2; findings on nonneoplastic lesions
 

are summarized in Appendix C, tables Cl and C2.
 

There was a relatively high incidence of subcutaneous fibromas
 

occurring only in the dosed male rats while none occurred in the
 

dosed females or in the controls. The incidence is as follows:
 

Male Rats
 

Matched Low High
 

Control D ose Dose
 

Number of animals necropsied (20) (50) (49)
 

Integumentary System
 

Fibromas 0 3 (6%) 9 (18%)
 

Grossly, the fibromas ranged from 5 cm tc 9 cm in diameter and
 

were located at points along the axillary, thoracic, abdominal,
 

and inguinal regions. Microscopically, they consisted of layers
 

of well-differentiated fibroblastic cells separated by dense
 

bands of mature collagen. A fibroadenoma of the mammary gland
 

also occurred in a high-dose male. Fibromas are occasionally
 

20
 
encountered spontaneously in aging rats, but this incidence is
 

considered unusual. In addition, several other unusual tumors
 

were observed in this study. These included three tumors of the
 

brain and two of subcutaneous tissue: astrocytoma in 1/48 (2%)
 

high-dose males; glioblastoma multiforme in 1/48 (2%) high-dose
 

females; malignant reticulosis in 1/50 (2%) low-dose females; and
 

myxosarcoma in 2/49 (4%) high-dose females.
 

As shown in the summary tables, there were malignant tumors in
 

the dosed rats that did not occur in the controls. These tumors
 

appeared to be randomly distributed, and the isolated or low
 

incidence minimizes any possible significance. The remaining
 

tumors that occurred in the dosed and control rats are commonly
 

observed as spontaneous tumors in aging rats. In some cases the
 

incidences of these spontaneous tumors exceeded those of the
 

matched controls, but the overall incidence did not exceed the
 

expected figures for aging rats.
 

A variety of degenerative and inflammatory conditions of the type
 

usually encountered in aged rats were observed, but none was
 

attributed to the test compound.
 

In the judgment of the pathologist, there was an increased
 

incidence of subcutaneous fibromas in dosed male rats under the
 

conditions of this bioassay.
 

21
 
D. Statistical Analyses of Results (Rats)
 

Tables El and E2 in Appendix E contain the statistical analyses
 

of the incidences of those primary tumors that occurred in at
 

least two animals in one group and with an incidence of at least
 

5% in one or more than one group.
 

In male rats, the result of the Cochran-Armitage test for
 

positive dose-related trend in the incidence of fibromas of the
 

subcutaneous tissue is significant (P = 0.006). The Fisher exact
 

comparison of the incidences of these tumors in the high-dose
 

group and its vehicle-control indicates a probability level of
 

0.036, which is above the 0.025 level required by the Bonferroni
 

inequality when multiple comparison is considered.
 

In female rats, a significant dose-related trend in the negative
 

direction (P = 0.043) is observed in the incidence of endometrial
 

stromal polyps of the uterus, where the incidence in the control
 

group exceeds those in the dosed groups; this significant
 

negative trend cannot, however, be accounted for by differential
 

survival.
 

22
 
IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

Mean body weights of the dosed groups of male mice were
 

comparable to those of the vehicle controls throughout the
 

bioassay, while those of the dosed groups of female mice were
 

lower, with a dose-related depression (figure 3). Fluctuation in
 

the growth curve may be due to mortality; as the size of a group
 

diminishes, the mean body weight may be subject to variation. No
 

other clinical signs related to administration of the trimethylphosphate were recorded.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female mice administered trimethylphosphate
 

by gavage at the doses of this bioassay, together with those of
 

the vehicle controls, are shown in figure 4.
 

The results of the Tarone test for positive dose-related trend in
 

mortality are not significant in either sex. At least 70% of the
 

male mice (39/49 [80%] in the high-dose group, 44/50 [88%] in the
 

low-dose group, and 14/20 [70%] in the matched controls) and at
 

least 59% of the female mice (29/49 [59%] in the high-dose group,
 

31/50 [62%] in the low-dose group, and 18/20 [90%] in the matched
 

23
 
S&OQOQQ Q OD 

Q Q 8 

A

 S E A A" AA° 

O Q 

A 30 n O 

Q 

O 

MALE MICE 

n VEHICLE CONTROL 

O LOW DOSE 

A HIGH DOSE 

0 I 1 1 1 1 1 T— 

0 10 20 30 40 50 60 70 

TIME ON STUDY (WEEKS) 

DDQ D naan D 

n D ° o o o o 

D D OOOOO O 

I­I 

a D r> p R A A 

A ^ O Q O 

O n

FEMALE MICE 

D VEHICLE CONTROL 

O LOW DOSE 

A HIGH DOSE 

TIME ON STUDY (WEEKS) 

Figure 3. Growth Curves for Mice Administered Trimethylphosphate by Gavage 

24 
PROBABILITYOF SURVIVAL PROBABILITY OF SUR

CQ C C9 C3 C3 C3 C) C 9 e » f > e3 

C i tJ i5 ?; §S !S I; i= S§ C; s', SS £! 8S 

 C

1 o j ( » 0 J 

1 s e '

> 0 > 0 D 

I 1- < m Q § i S 5 9 *** * 0 8- 0 § 0 > s - r~ f— (3 O P S m m

2 m m n S

-4* o " 1 0 O o rn g- rICE ONTROt 

H fc 

i i*- —^

>• m

eL o 0

N> 3 Z «. z Cn g. g o -1

C 5' = 

m

i? o o

5 If 8­ C <

—1 m a m* -J j ^_ 3 tn ° » s- CD — 

<• 9- j

ts § - -&'^!

 — §- 

S ^gP-J|^ 

6

Y r*^ § 

8­ 1 " fc 

T-D— t 

0 8- s­ c 

^ | 

« i­ ^ 
controls) lived to the end of the study, providing adequate
 

numbers of animals of each sex at risk for development of late-


appearing tumors.
 

C. Pathology (Mice)
 

Histopathologic findings on neoplasms in mice are summarized in
 

Appendix B, tables Bl and B2; findings on nonneoplastic lesions
 

are summarized in Appendix D, tables Dl and D2.
 

A high incidence of endometrial adenocarcinomas and a few other
 

types of malignant uterine tumors occurred only in the female
 

mice administered trimethylphosphate, as shown in the following
 

tabulation:
 

Female Mice
 

Matched Low High
 

Control Dose Dose
 

Number of animals with tissues
 

examined microscopically (16) (40) (37)
 

Uterus/Endometrium
 

Adenocarcinoma, NOS* 0 7(18%) 13(35%)
 

Squamous-cell carcinoma 1(3%)
 

Uterus
 

Leiomyosarcoma 0 1(3%) 0
 

*Not otherwise specified
 

Grossly, the uterine tumors were masses 1 cm to 2 cm in diameter,
 

which were usually limited to one horn. Microscopically, the
 

26
 
majority of the tumors appeared to arise from the endometrium as
 

irregular acinar structures with slit-like lumens that were
 

composed of flat to low cuboidal hyperchromatic epithelial cells.
 

The neoplastic glandular structures widely invaded the myometrium
 

and often extended to the serosa. There was a scirrhous reaction
 

in some areas in which tumors were involved. The remainder of
 

the tumors appeared to arise from endometrial polypoid structures
 

that contained columnar shaped cells with high nuclear/cytoplasmic ratios and numerous mitoses. A few of these formed papillary
 

structures and had cystic areas. Overall, the uterine adenocarcinomas appeared to be highly malignant. There was vascular
 

involvement and pulmonary metastases in one low-dose and four
 

high-dose mice. The tumors appeared to be more aggressive in the
 

high-dose animals, since metastases frequently occurred in those
 

groups.
 

Spontaneous adenocarcinomas of the uterus are uncommon in mice.
 

Their high incidence in this study is attributed to the
 

administration of trimethylphosphate.
 

There were interstitial-cell tumors of the testes in two low-dose
 

male mice, which consisted of sheets of basophilic, round to polygonal cells that separated and displaced seminiferous tubules.
 

These tumors also do not occur commonly in mice.
 

27
 
There were a few malignant or unusual tumors that occurred in the
 

dosed mice, which were not seen in the controls. In the male
 

mice, these included a rhabdomyosarcoma (one low-dose), a gastric
 

squamous-cell carcinoma (one high-dose), and an adenocarcinoma of
 

the lacrimal gland (one high-dose). In the female mice, there
 

was an osteosarcoma that metastasized to the lung and kidney (one
 

low-dose), an oligodendroglioma of the brain (one low-dose), an
 

ameloblastoma of the mandible (one high-dose), and an
 

arrhenoblastoma (Sertoli-cell tumor) of the ovary (one high-


dose). Although these represent an unusual array of tumors,
 

little significance was attributed to them because of their
 

apparent sporadic and low incidence. Other tumors observed
 

included those of hepatocellular and hematopoietic origin that
 

had equivalent frequencies in the control and dosed mice.
 

There were two nonneoplastic changes that appeared to be associated with the uterine tumors described above. One was hydronephrosis; four of the six mice with this lesion had endometrial
 

adenocarcinomas, and one had a uterine leiomyosarcoma. Although
 

involvement of the urinary tract was microscopically evident in
 

only two cases, it is likely that obstruction by the tumor
 

occurred in the other cases at some point along the urinary
 

tract. The other change was extensive thrombosis of the pulmo28
 
nary arteries that occurred in three high-dose mice with
 

pulmonary metastases of the endometrial adenocarcinomas.
 

There were several other nonneoplastic changes considered to be
 

either spontaneous or intercurrent disease processes that are
 

common in mice on long-term studies.
 

In the judgment of the pathologist, it was concluded that because
 

of the high incidence and apparent dose response of endometrial
 

adenocarcinomas in the dosed female mice, trimethylphosphate was
 

clearly carcinogenic in female B6C3F1 mice under the conditions
 

of this bioassay.
 

D. Statistical Analyses of Results (Mice)
 

Tables Fl and F2 in Appendix F contain the statistical analyses
 

of the incidences of those primary tumors that occurred in at
 

least two animals in one group and with an incidence of at least
 

5% in one or more than one group.
 

In male mice, neither the result of the Cochran-Armitage test for
 

positive dose-related trend in incidences of tumors nor the
 

result of the Fisher exact test for direct comparison of
 

incidences of tumors in dosed groups with those in the controls
 

is significant.
 

In females, the result of the Cochran-Armitage test for the
 

29
 
incidence of adenocarcinoma of the uterus/endometrium is
 

significant (P = 0.003) and the Fisher exact test shows that the
 

incidence of the tumors in the high-dose group is significantly
 

higher (P = 0.004) than that in the controls. No occurrence of
 

such tumors has been reported in the laboratory historical
 

controls, which consist of 100 female B6C3F1 mice. The
 

statistical conclusion suggests that this incidence of tumors in
 

female mice is related to the administration of the test
 

chemical.
 

The incidence of lung tumors in the low-dose group of female mice
 

is significantly lower (P = 0.023) than that in the controls
 

(controls 3/20 [15%], low-dose 0/48). This significance in the
 

negative direction cannot be accounted for by the fact that the
 

survival in the low-dose group (62%) is lower than that in the
 

controls (90%), since the high-dose group has a survival (59%)
 

comparable to that of the low-dose group yet has an incidence of
 

tumors (6/45 [13%]) that is not substantially different from that
 

of the controls.
 

30
 
V. DISCUSSION
 

Under the conditions of this bioassay, mean body weights of the
 

dosed male and female rats and of the dosed female mice were
 

slightly lower than those of corresponding vehicle controls, with
 

dose-related depression in the weights, while those of the dosed
 

male mice were essentially unaffected. Survival of all groups of
 

animals was high, and adequate numbers of both rats and mice were
 

at risk for development of late-developing tumors.
 

In the male rats, the incidence of fibromas of the subcutanous
 

tissue was significantly higher (P = 0.036) in the high-dose
 

group than in the vehicle controls (control 0/20, low-dose 2/50,
 

high-dose 9/49), and there was a significant dose-related trend
 

(P = 0.006) in the incidences of these fibromas. In the female
 

rats, no tumors occurred in the dosed groups at significantly
 

increased incidences.
 

In the male mice, no tumors were observed in the dosed groups at
 

significant incidences, compared with the vehicle controls. In
 

the female mice, the incidence of adenocarcinomas of the
 

endometrium was significantly higher (P = 0.004) in the high-dose
 

group than in the vehicle controls (controls 0/16, low-dose 7/40,
 

high-dose 13/37), and there was a significant dose-related trend
 

(P = 0.003) in the incidences of these adenocarcinomas. None of
 

31
 
these tumors occurred among the 100 historical-control female
 

B6C3F1 mice at this laboratory. In addition, a squamous-cell
 

carcinoma occurred in the uterus of a high-dose female mouse and
 

a leimyosarcoma in the uterus of a low-dose female mouse.
 

No significant gonadal effects were found in either rats or mice.
 

No long-term toxicity studies with tritnethylphosphate have been
 

reported. In subacute experiments with rabbits, Deichmann and
 

Witherup (1946) observed tremors and a flaccid paralysis after 6
 

days of oral gavage at 300 mg/kg body weight.
 

It is concluded that under the conditions of this bioassay,
 

trimethylphosphate was carcinogenic in female B6C3F1 mice,
 

inducing adenocarcinomas of the uterus/endometrium.
 

Trimethylphosphate was associated with the induction of benign
 

fibromas of the subcutaneous tissue in male Fischer 344 rats. No
 

evidence of carcinogenicity of the compound was obtained in
 

female rats or in male mice.
 

32
 
VI. BIBLIOGRAPHY
 

Adler, I. D., Ramarao, G., and Epstein, S. S., In vivo
 

cytogenetic effects of trimethylphosphate and of TEPA on
 

bone marrow cells of male rats. Mutation Res. 13;263-273,
 

1971.
 

Aldrich Library of Infrared Spectra, 2nd Ed., Aldrich Chemical
 

Co., Inc., 1976.
 

Armitage, P., Statistical Methods in Medical Research, John Wiley
 

& Sons, Inc., New York, 1971, pp. 362-365.
 

Barnard, P. W. C., Bunton, C. A., Llewellyn, D. R. , Vernon, C.
 

A., and Welch, V. A., The reactions of organic phosphates.
 

Part V. The hydrolysis of triphenyl and trimethyl
 

phosphates. J. Chem. Soc. , 1961, pp. 2670-2676.
 

Berenblum, I., ed., Carcinogenicity Testing, A Report on the
 

Panel on Carcinogenicity of the Cancer Research Commission
 

of the UICC, Vol. 2, International Union Against Cancer,
 

Geneva, 1969.
 

Billman, J. H. , Radike, A., and Mundy, B. W., Alkylation of
 

amines. I. J,. Amer. Chem. Soc. 64:2977-2978, 1942.
 

Chem Sources ^ USA, Directories Publishing Co., Inc., Flemington,
 

N.J., 1977.
 

Cox, D. R., Regression models and life tables. J. R. Statist.
 

Soc. B 34(2);187-220, 1972.
 

Cox, D. R., Analysis of Binary Data, Methuen & Co., Ltd., London,
 

1970, pp. 48-52.
 

Deichmann, W. B. and Witherup, S. , Observations on the effects of
 

trimethylphosphate upon experimental animals. J. Pharmacol.
 

Exp. Ther. 88;338-342, 1946.
 

Epstein, S. S., Bass W., Arnold, E., and Bishop, Y. , Mutagenicity
 

of trimethylphosphate in mice. Science 168:584-586, 1970.
 

Gart, J. J., The comparison of proportions: a review of
 

significance tests, confidence limits and adjustments for
 

stratification. Rev. Int. Statist. Inst. 39:148-169, 1971.
 

33
 
Harbison, R. D., Diwivedi, C., and Evans, M. A., A proposed
 

mechanism for trimethylphosphate-induced sterility.
 

Toxicol. Appl. Pharmacol. 35:481-490, 1976.
 

Hinkamp, J. B. and Warren, J. A., Surface ignition control by
 

phosphorus fuel additives. Indust. Engineering Chem.
 

5^:251-256, 1958.
 

Kaplan, E. L. and Meier, P., Nonparametric estimation from
 

incomplete observations. J. Amer. Statist. Assoc.
 

53_:457-481, 1958.
 

Linhart, M. S., Cooper, J., Martin, R. L., Page, N., and Peters,
 

J., Carcinogenesis bioassay data system. Comp. and Biomed.
 

Res. _7_:230-248, 1974.
 

Matsunaga, H., Okumura, 1., Takada, I., Japan pat. 74 17,655 

(1974 to Sumitomo Chemical Co., Ltd.) Chem. Abst. 82. 

44412h, 1975. 

Miller, R. G. , Jr. , Simultaneous Statistical Inference,
 

McGraw-Hill Book Co., New York, 1966, pp. 6-10.
 

Newell, G. W., Jorgenson, T. A., Rushbrook, C. J. , and Spanggord,
 

R., Preliminary Investigations of new orally active
 

reference mutagens. TAP, 1977, p. 108.
 

Saffiotti, U., Montesano, R., Sellakumar, A. R., Cefis, F., and
 

Kaufman, D. G. , Respiratory tract carcinogenesis in hamsters
 

induced by different numbers of administrations of benzo(a)
 

pyrene and ferric oxide. Cancer Res. 32:1073-1081, 1972.
 

Spanggord, R. J., Stanford Research Institute, Memorandum dated
 

October 12, 1977.
 

Tarone, R. E., Tests for trend in life table analysis.
 

Biometrika 62(3);679-682, 1975.
 

Toy, A. D. F., Preparation of tetramethyl pyrophosphate. J.
 

Amer. Chem. Soc. ^1:2268-2269, 1949.
 

34
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS ADMINISTERED TRIMETHYLPHOSPHATE
 

BY GAVAGE
 

35
 

TABLE A1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS 

ADMINISTERED TRIMETHYLPHOSPHATE BY GAVAGE 

VEHICLE LOW DOSE 

CONTROL 

ANIMALS INITIALL Y IN STUD Y 20 50 

ANIMAL S NECROPSIED 20 50 

ANIMALS EXAMINE D HI STOP ATHO LOGICALLY 20 50 

INTEGUMENTARY SYSTEM 

*SKIN 

SEBACEOUS ADENOMA 

FIBROMA 

(20) (50) 

1

1

 (2%) 

 (2%) 

*SUBCUT TISSUE 

NEOPLASM, NOS 

FIBROMA 

LIPOMA 

L2IOMYOSARCOMA 

HtMANGIOSAECOMA 

(20) 

1 (555) 

(50) 

2

1

1

 (4X) 

 (2%) 

 (2%) 

RESPIBAT08Y SYSTEM
 

#LUNG (19) (49)
 

ALVEOLAR/EEONCHIOLAR ADENOMA
 

ALVEOLAR/BBONCHIOLAR CARCINOMA 2 (4%)
 

LEIOMYOSARCOMA, METASTATIC 1 (2%)
 

HEMATOPOIETIC SYSTEM
 

•MULTIPLE ORGANS (20) (50)
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 (2%)
 

LEUKEMIA,NCS 3 (15%) 4 (8%)
 

UNDIFFEHENTIATED LEUKEMIA 5 (25%) 15 (30%)
 

LYMPHOCYTIC LEUKEMIA
 

tLYMPH NODE (20) (46)
 

OSTEOSARCCMA, METASTATIC 1 (2%)
 

#LUNG (19) (49)
 

LEUKEMIA,NOS
 

CIRCULATORY SYSTEM
 

__IQN£_
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

3 50 ANIMAL S WER E INITIALL Y IN THE STUDY , BUT ONE ANIMA L WAS FOUN D

IN A MAL E GROUP . 

HIGH DOSE 

350 

49 

1*9 

(49)
 

(49)
 

1 (2%)
 

9 (18%)
 

(46)
 

4 (9%)
 

1 (2%)
 

(49)
 

8 (16%)
 

15 (31%)
 

1 (2%)
 

(46)
 

(46)
 

1 (2%)
 

 TO EE A FEMAL E 

37 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

VEHICLE LOW DOSE HIGH DOSE 

CONTROL 

DIGESTIVE SYSTEM 

tLIVER (20) (48) (48) 

NEOPLASTIC NODULE 1 (2%) 1 (2%) 

'OSTEOSARCOHA, HETASTATIC 1 (2%) 

#COLON (12) (41) (43) 

ADENOCARCINOHA , NOS 1 (2%) 

*RECTOB (20) (50) (49) 

ADENOCARCINOMA , NOS 1 (2X) 

URINAfi Y SYSTEM 

#KIDNE Y (20) (49) (48) 

ADENOMA, NOS 1 (2X) 

TUBULAR-CEL L ADENOCARCINOBA 1 (2%) 

LIPOSARCCM A 1 (2%) 

OSTEOSARCCBA, METASTATIC 1 (2%) 

tURINAR Y BLAEDE R (11) (36) (35) 

THANSITIONAL-CELL CARCINOM A 1 ( 3 % ) 

ENDOCRINE SYSTE M 

*PITUITARY (16) (44) (38)
 

CHROMOPHOEH ADENOMA 4 (25%) 13 (30X) 8 (21X)
 

#ADRENAL (20) (48) (47)
 

CORTICAL JtENOBA 1 (2X)
 

PHEOCHROMOCYTOHA 1 (5X) 4 (8X) 7 (15X)
 

tTHYROID (19) (45) (46) 

FOLLICULAR-CELL CARCINOM A 2 (4X) 

C-CELL ADENOM A 1 (5X) 3 (7X) 

C-CELL CABCINOB A 2 (4X) 

#PANCREATIC ISLETS (18) (44) (45) 

ISLET-CELL ADENOM A 1 (6X) 1 (2X) 1 (2X) 

ISLET-CELL CARCINOM A 1 (2X) 

REPRODUCTIVE SYSTEM
 

*MAMMAR Y GLANt (20) (50) (49)
 

FIBROADENOgA
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

38
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

VEHICLE

CONTROL 

 LOW DOSE HIGH DOSE 

*PREPOTIAL GLAND

ADENOMA , NCS

 (20)

 1 (5%) 

 (50) (49) 

(tTESTIS

INTERSTITIAL-CELL TUHOR

 (16)

 11 (69X)

 (46)

 33 (72*)

 (46) 

 25 (54S) 

NERVOU S SYSTEM 

#BRAIN

ASTROCYTCMA

 (20) (50) (48) 

1 (2%) 

SPECIAL SENSE CRGANS 

NONE 

MUSCULOSKELETAL SYSTEM 

NON£ 

BODY CAVITIES 

*ABDOMINAL CAVITY

OSTEOSARCOMA, UNC PRIM OR META

 (20) (50)

 1 (2%) 

 (49) 

*TUNICA VAGINALIS

MESOTHELICMA, NOS

 (20) (50)

 2 (4»)

 (49) 

1 (2%) 

ALL OTHER SYSTEMS 

NONE 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY

NATURAL DEATHS

MORIBUND SACRIFICE

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE

ANIMAL MISSING 

ANIMAL DELETED (WRONG SEX) 

 20

 4

 8

 8

 50 

 11 

 11 

 28 

50 

9 

23 

17 

1 

^IN£JLUPJS_A.UTJ:LJZED_ ANIBAL.JJ 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY * NUMBER OF ANIMALS NECROPSIED 

39
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

VEHICLE LOW DOSE HIGH DOSE
 

CONTROL
 

r UHOR SUMMAR Y
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 19 U8 U3
 

TOTAL PRIMARY TUMORS 28 91 92
 

TOTAL ANIMALS WITH BENIGN TUMORS 15 41 33
 

TOTAL BENIGN TUMORS 19 61 55
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 8 24 29
 

TOTAL MALIGNANT TUMORS 9 26 3U
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 2
 

TOTAL SECONDARY TUMORS 4
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT 3 3
 

TOTAL UNCERTAIN TUMORS J 3
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC 1
 

TOTAL UNCERTAIN TUMORS 1
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

# SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

40
 
TABLE A2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

ADMINISTERED TRIMETHYLPHOSPHATE BY GAVAGE

 

ANIMAL S 

ANIMAL S 

ANIMAL S 

INITIALL Y IN STUDY 

NECBOPSIE D 

EXAMINE D HIS1 rOPATHO LOGICALLY 

INTEGUMENTABY SYSTEM
 

*SUBCUT TISSUE
 

SQUAMOUS CELL CARCINOMA
 

KERATOACANTHOMA
 

MYXOSARCCMA
 

LIPOMA
 

OSTEOSARCOMA
 

NEUROFIBBCMA
 

RESPIRATORY SYSTEM
 

#LUNG
 

SG.UAMOUS CELL CARCINOMA, METASTA
 

ALVEOLAR/BRONCHIOLAB ADENOMA
 

ALVEOLAR/BECNCHIOLAR CARCINOMA
 

FIBROSARCCPA, INVASIVE
 

OSTEOSARCCKA, METASTATIC
 

HEMATOPOIETIC SYSTEM
 

f BRAI N 

MALIGNAN T BETICULOSIS 

*MULriPLE ORGANS
 

MALIGNANT LYMPHOMA, NOS
 

LEUKEMIA,NCS
 

UNDIFFERENTIATED LEUKEMIA
 

#SPL£EN
 

SARCOMA, NOS
 

CIRCULATORY SYSTEM
 

VEHICLE LOW DOSE HIGH DOSE 

CONTROL 

20 50 350 

20 50 49 

20 50 48 

(20) (50) (49) 

1 (2%) 

1 (2«) 

2 (4X) 

1 (5%) 

1 (2») 

1 (2X) 

(20) (50) (45) 

1 (2%) 

2 (4X) 

1 (2%) 

1 (2%) 

1 (2*) 

(20) (50) (48) 

1 (2%) 

(20) (50) (49) 

1 (2%) 

2 (10X) 4 (8%) 1 (2%) 

1 (5%) 9 (18%) 11 (22%) 

(20) (U9) (46) 

1 (5%) 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

a 50 ANIMALS WERE INITIALLY IN THE STUDY, BUT ONE ANIMAL HAS FOUND TO EE A MALE
 

IN A FEMALE GROUP.
 

41
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

VEHICLE LOW DOSE HIGH DOSE 

CONTROL 

DIGESTIVE SYSTE M 

NON E 

UHINAB Y SYSTE M 

#URINAR Y BLADDER (17) (32) (31)

 

TfiANSITIONAL-CELL CARCINOM A 1 (6*)

 

ENDOCRIN E SYSTE M 

fPITUITARY (20) (48) (41) 

CHROMOPHOE E ADENOMA 9 (USX) 21 (44X) 18 (<*«*) 

# ADRENA L (19) (48) (48) 

COPTICAL ADENOMA 1 (2*) 

PHEOCHROMOCYTOM A 2 (4*) 1 (2X) 

tTHYfiOID (19) (47) (43) 

FOLLICULAR-CELL CARCINOMA 1 (2*) 

C-CELL ADENOM A 6 (13*) 3 (7*) 

C-CELL CARCINOM A 2 (11*) 2 (5X) 

tPARATHYROID (10) (28) (23)

 

CHIEF-CELL ADENOM A 1 (4*)

 

REPRODUCTIVE SYSTEM 

*MABMAR Y GLAN E (20) (50) (49) 

ADENOCARCINOMA, NOS 1 (5*) 

FIBROADENOBA 2 (10*) 3 (6*) 5 (10*) 

tUTERUS (20) (45) (44)

 

ENDOMETBIAL STHOMAL POLYP 2 (10*) 1 (2*) 

*CERVIX UTERI (20) (45) (44) 

LEIOMYOSARCOBA 1 (2*) 

NERVOUS SYSTEM
 

IBRAIN (20) (50) (48) 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECHOPSIED
 

42
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

VEHICLE LOW DOSE HIGH DOSE 

CONTROL 

SPECIAL SENS E ORGAN S 

*ZY«BAL'S GLAND (20) (50) (149)
 

SEBACEOUS ADENOMA 1 (2%)
 

MUSCULOSKELETAL SYSTEM
 

*STEKNUM (20) (50) («9)
 

FIBBOSAECOMA 1 (2%)
 

BODY CAVITIES
 

NONE
 — _ _ _«_ «__ .._ _ _ _ _._ ______ _ — — —. — -.— —•____•_•_.•—_•_...•__•..„_..• — •••— •——• — •— ——•— ••• — — — —— — — — — «
 

A LL OTHER SYSTEM S 

NON E 

ANIMA L DISPOSITION SUMMAR Y 

ANIMALS INITIALLY IN STUDY 20 50 50
 

NATURA L DEATHS 10
 

MORIBUN D SACRIFICE 12
 

SCHEDULED SACRIFICE 

ACCIDENTALL Y KILLED 

TERMINA L SACRIFICE 12 36 27
 

ANIMA L MISSING 

ANIMA L DELETED(WRON G SEX) 1
 

l_IfiCjiDDES_AUTCLYZED_ANIB115­

* NUMBER OP ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECROPSIED
 

43
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

VEHICLE


CONTROL
 

TUMOR SUMMARY 

TOTAL ANIMALS WITH PRIMARY TUMORS* TOTAL PRIMARY TUMORS 15

 22

TOTAL ANIMALS WITH BENIGN TUMORS

TOTAL BENIGN TUMORS

 12

 14

TOTAL ANIMALS WITH MALIGNANT TUMORS

TOTAL MALIGNANT TUMORS

 8

 8

TOTAL ANIHALS HITH SECONDARY TUMORS* TOTAL SECONDARY TUMORS

TOTAL ANIHALS WITH TUMORS ONCERTAIN­ BENIGN OR MALIGNANT 

TOTAL UNCERTAIN TUMORS 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OH HETASTATIC 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

 LOW DOSE HIGH DOSE 

 40 42 

 51 54 

 28 29 

 34 33 

 17 19 

 17 21 

1 2 

1 2 

# SECONDARY TUBORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE ADMINISTERED TRIMETHYLPHOSPHATE
 

BY GAVAGE
 

45
 

TABLE B1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

ADMINISTERED TRIMETHYLPHOSPHATE BY GAVAGE 

VEHICLE LOW DOSE 

CONTROL 

ANIMALS INITIALL Y IN STUD Y 20 50 

ANIMALS NECROESIED 20 50 

ANIMAL S EXAMINE D HIS! 'OPATHO LOGICALLY 20 50 

INTEGUMENTARY SYSTEM
 

*SKIN (20) (50)
 

RHABDONYOSAHCOMA 1 (2%)
 

*SUBCUT TISSUE (20) (50)
 

HEMANGIOSABCOMA
 

ANGIOSARCOKA 1 (5%)
 

RESPIfiATORY SYSTEM
 

fLUNG (20) (50)
 

ADENOCARCINOMA, NOS, METASTATIC
 

Hi-PATOCELLULAR CARCINOMA, METAST 3 (6%)
 

ALVEOLAR/BRONCHIOLAR ADENOMA 1 (5%) 3 (6X)
 

ALVEOLAR/BEONCHIOLAR CARCINOMA 2 (10%) 8 (16%)
 

SARCOMA, NCS, METASTATIC
 

ANGIOSARCCKA, METASTATIC 1 (5%)
 

HEMATOPOIETIC SYSTEM
 

*«OLTIPL5 ORGANS (20) (50)
 

MALIGNANT LYMPHOMA, NOS 1 (2%)
 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE 1 (5%)
 

MALIGNANT LYMPHOMA, MIXED TYPE 1 (2%)
 

UNDIFFERENTIATED LEUKEMIA 1 (2%)
 

#SPLiEN (18) (45) 

HEMANGIOH A 1 (6% ) 

tMEDIASTINA L L.NODE (18) (45) 

HEPATOCELLULA R CARCINOMA , METAST 1 (2%) 

*MESENTERIC L. NODE (18) (45)
 

M ALIGN ANT^YMPHOMA^NQS ______________
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE 

49 

49 

49 

(49)
 

(49)
 

1 (2%)
 

(49)
 

1 (2%)
 

2 (4%)
 

6 (12%)
 

3 (6%)
 

1 (2%)
 

(49)
 

2 (4%)
 

1 (2%)
 

2 (4%)
 

1 (2%)
 

(44)
 

(41)
 

(41)
 

47
 
TABLE B1. MALE MICE : NEOPLASMS (CONTINUED) 

VEHICLE LOW DOSE HIGH DOSE
 

CONTROL
 

MALIGNANT LYMPHOHA. HIKED TYPE 1 (2%) 1 (2%)
 

*SMALL INTESTINE {20) (49) (45)
 

MALIGNANT IYHPHOHA. NOS 2 (4S)
 

MALIGNANT LYHPHOHA, HIXED TYPE 1 (5%)
 

#COLON (20) (48) (47)
 

MALIGNANT IYMPHOHA, NOS 1 (2%)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

fLIVER (20) (48) (49)
 

ADENOCARCINOMA, NOS 1 (2%)
 

HEPATOCELLULAR ADENOHA 1 (2%)
 

HEPATOCELLULAR CARCINOMA 4 (20%) 9 (19S) 8 (16X)
 

• STOMACH (18) (50) (46)
 

SQUAMOUS CELL CARCINOMA 1 (2%)
 

URINARY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

*ADRENAL (17) (48) (43)
 

PhEOCHROMOCYTOMA 1 (6X)
 

*THYROID (9) (35 ) (39) 

FOLLICULAR-CEL L ADENOMA 1 (11X ) 1 (3«) 

tPANCREATIC ISLETS (20) (46) (42) 

ISLET-CELL ADENOM A 1 (2«) 

REPRODUCTIV E SYSTEM 

*TESTIS (20) (45) (47) 

INTE£STITIAL-CE.LL._TUMflR. 

# NUMBE R OF ANIMAL S WITH TISSUE EXiHIHED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

48 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

VEHICLE

CONTROL 

 LOW DOSE HIGH DOSE 

NERVOU S SYSTEM 

NONE 

SPECIAL SENSE ORGAN S 

*EYE/LACRIHA L GLAN D

ADENOCARCINCMA, NOS

 (20) (50) (U9 ) 

1 (2%) 

MUSCULOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

*ABDOMINA L CAVITY

SARCOMA, NOS

LIPOMA

 (20) (50) (<*9) 

1

 1

 (2«) 

 (2S) 

ALL OTHER SYSTEMS 

NON E 

ANIMA L DISPOSITION SUMMAR Y 

ANIMALS INITIALLY IN STUDY

NATURA L DEATHS

HORIBUN D SACRIFICE

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINA L SACRIFICE

ANIMA L MISSING 

 20

 5 

1

 14

 50

3 

3

 <*«

 «9 

9 

1 

 39 

t NUMBER

* NUMBER

 OF

 OF

 ANIMAL S

 ANIMALS

 HITH TISSUE

 NECROPSIED 

 EXAMINE D MICROSCOPICALLY 

49 
TABLE B1. MALE MICE : NEOPLASMS (CONTINUED) 

VEHICLE LOW DOSE HIGH DOSE
 

CONTROL
 

1UHOE SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 11 26 26
 

TOTAL PRIMARY TUMORS 1i» 31 32
 

TOTAL ANIMALS WITH BENIGN TUMORS U 8 7
 

TOTAL BENIGN TUMORS u 8 7
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 9 20 22
 

TOTAL MALIGNANT TUMORS 10 23 25
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 1 3 3
 

TOTAL SECONDARY TUMORS 1 4 4
 

TOTAL ANIMAL S WITH TUMORS UNCERTAIN ­ BENIGN O R MALIGNAN T

 

TUTAL UNCERTAI N TUMORS 

TOTAL ANIMAL S WIT H TUMORS UNCERTAIN ­ PRIMAR Y OR METASTATIC

 

TOTAL UNCEBTAI N TUMORS 

* PRIllARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS 

# SECONDAR Y TUMORS : METASTATIC TUMORS 08 TUHORS INVASIVE INTO AN ADJACENT ORGA N 

50 
TABLE B2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

ADMINISTERED TRIMETHYLPHOSPHATE BY GAVAGE

 

VEHICLE LOW DOSE 

CONTROL 

ANIMALS INITIALL Y IN STODY 20 50 

ANIMAL S HISSING 

ANIMAL S NECROPSIED 20 50 

ANIMAL S EXAMINE D HIS! rOPATHOLOGICALLY 20 50 

INTEGUMENTAR Y SYSTEM 

*SUBCUT TISSUE (20) (50)

SARCOMA, NOS 1 (5%) 

BESPIEAT08Y SYSTEM 

#LUN G (20) (48)

NEOPLASM, NOS, UNC PRIM OR META 1 (2X)
 

CARCINOMA, NOS, UNC PRIM OR META 1 (5X)
 

SQUAMOOS CELL CARCINOMA, METASTA
 

TBANSITIONAl-CELL CARCINOMA, MET 1 (5X)
 

ADENOCABCINOMA, NOS, METASTATIC
 

HiPATOCEIIULAR CARCINOMA, METAST 1 (2X)
 

ALVEOLAR/BRONCHIOLAR ADENOMA 2 (10*)
 

ALVEOLAR/ERONCHIOLAR CARCINOMA 1 (5X)
 

SARCOMA, NCS, METASTATIC 1 (5X)
 

LEIOMYOSARCOMA, METASTATIC 1 (2%)
 

03TEOSARCOMA, METASTATIC 1 (2X)
 

HEMATOPOIETIC SYSTEM 

*MULTIPLE ORGAN S (20) (50)

MALIGNAN T LYMPHOMA , NOS 1 (5X) 3 (6X)

MALIG.LYMPHOMA , LYMPHOCYTIC TYPE 1 (2X) 

MALI3.LYMPHOMA , HISTIOCYTIC TYPE 1 (2X) 

MALIGNAN T LYMPHOMA , MIXE D TYPE 1 (5«) 4 (8%)

UNDIFFEBENIIATE D LEUKEMI A 1 (2X)

LYBPHOCYTIC LEUKEMI A 1 (5X) 1 (2X)

GRANULOCYTI C SARCOMA 1 (2X) 

#SPLEEN (17) (U8)

M ALIGN ANUYMPHOMA^.MIXED.TYPE J_J2Sl 

# NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

3 50 ANIMAL S HERE INITIALL Y IN THE STUDY, BUT ONE ANIMA L WAS FOUND

IN A FEMAL E GROUP . 

HIGH DOSE 

950 

2 

HI 

HI 

 (47) 

 (45) 

1 (2%)
 

4 (9%)
 

5 (11X)
 

1 (2X)
 

 (1*7) 

2 (4X) 

2 (4K ) 

2 (UX ) 

2 (UX ) 

 (UU ) 

 TO EE A MAL E 

51 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

VEHICLE LOW DOSE
 

CONTROL
 

#MANDIBULAE L. NODE (17) (46)
 

MALIGNANT LYMPHOMA, NOS
 

#LYMPH NODE OF THORAX (17) (46)
 

ADENOCARCINOMA, NOS, METASTATIC
 

#MEDIASTINAL I. NODE (17) (46)
 

ADENOCARCINOMA, NOS, METASTATIC
 

MALIGNANT LYMPHOMA, MIXED TYPE 1 (2*)
 

fMESENTERIC L. NODE (17) (46)
 

MALIGNANT LYMPHOMA, MIXED TYPE 1 (2%)
 

#RENAL LYMPH NODE (17) (46)
 

ADENOCARCINCMA, NOS, METASTATIC
 

#LIVER (20) (50)
 

MALIGNANT LYMPHOMA, MIXED TYPE 1 (5X)
 

#KIDNEY (19) (50)
 

MALIGNANT LYMPHOMA, NOS 1 (5%)
 

#THYMUS (2) (1)
 

MALIGNANT LYMPHOMA, NOS
 

CIRCULATORY SYSTEM
 

""LYMPHATICS CF UTERUS (20) (50)
 

S^UAMOUS CELL CARCINOMA, INVASIV
 

ADENOCARCINCMA, NOS, INVASIVE
 

*PULi10NARY ARTERY (20) (50)
 

ADENOCARCINOMA, NOS, INVASIVE
 

*UTERINE VEIN (20) (50)
 

SJUAMOUS CELL CARCINOMA, INVASIV
 

ADENOCARCINOMA, NOS, INVASIVE 1 (2X)
 

DIGESTIVE SYSTEM
 

#LIVER (20) (50)
 

HEPATOCELLULAR ADENOMA 1 (5XL 2 ( 4%)
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECHOPSIED
 

HIGH DOSE
 

(43)
 

1 (2%)
 

(13)
 

1 (2%)
 

(13)
 

1 (2X)
 

(13)
 

1 (2«)
 

(13)
 

1 (2%)
 

(44)
 

(44)
 

(D
 

1 (100S)
 

(47)
 

1 (2%)
 

1 (2%)
 

(47)
 

2 (4%)
 

(17)
 

1 (2%)
 

(44)
 

52
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

DOSE 

(2X) 

(5X) 

(2X) 

(4X) 

(3X) 

(3X) 

(3X) 

(35X) 

C«X) 

(4X) 

HEPATOCELLULA E CARCINOMA 

HEMANGIOSABCON A 

fPANCREA S 

ADENOCARCINOMA , NOS, INVASIVE 

ISTOflACH 

ADENOBATOU S POLYP, NOS 

UBINAR Y SYSTE M 

#KIDNE Y 

TRANSITIONAL-CELL CARCINOMA 

ADENOCARCINOMA , NOS, HETASTATIC 

OSTEOSARCCHA, METASTATIC 

#URINAR Y BLADDER 

ADENOCARCINOMA ,

ENDOCRINE SYSTEM 

• ADRENA L 

ADENOCARCINOHA ,

REPRODUCTIVE SYSTEM 

tUTEROS 

 NOS, INVASIVE 

 NOS, INVASIVE 

TRANSITIONAL-CELL CARCINOMA. BET 

LEIOMYOSARCOMA 

ENDOMETRIA L STROHAL POLYP 

iUTERUS/ENDOHETR IUM 

SQOAMOUS CELL CARCINOMA 

ADENOCAECINOBA, HOS 

#OVARY 

SERTOLI-CELL TOHOH 

HEHANGIOM A 

NERVOUS SYSTEH 

# BRAIN 

OLIGODENDBOGLIOHA 

VEHICLE LOW DOSE HIGH 

CONTROL 

1 (5X) 2 (4%) 

1 

(20) (48) (42) 

2 

(17) (46) (47) 

1 (6%) 

(19) (50) (44) 

1 (5X) 

1 

1 (2X) 

(13) (27) (25) 

1 

(17) (46) (39) 

1 

(16) (40) (37) 

1 (6X) 

1 OX) 

2 (5X) 1 

(16) (40) (37) 

1 

7 (18«) 13 

(10) (26) (28) 

1 

1 

(19) (49) («6) 

1 (2X1 

* NUMBER OF ANIMALS BITH TISSUE EXABIMED HICECSCOPICALLI 

* NUMBER OF ANIMALS NECROPSIED 

53 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

VEHICLE

CONTROL 

 LOW DOSE HIGH DOSE 

SPECIAL SENSE ORGANS 

NON£ 

HUSCULOSKELETAL SYSTEM 

*BONE

OSTEOSARCCHA

*MANDIBLE

AMELOBLASTCMA

 (20)

 (20)

 (50)

 1 (2X) 

 (50)

 (U7) 

 («7) 

1 (2X) 

BODY CAVITIES 

*ABDOBINAL CAVITY

LIPOMA

^PERITONEUM

ADENOCARCINOHA, NOS, INVASIVE

 (20)

 (20)

 (50)

 1 (2%) 

 (50)

 (47) 

 (U7) 

1 (2X) 

ALL OTHER SYSTEMS 

ADIPOSE TISSDE 

SQUAMOUS CELL CARCINOMA, METASTA

ADENOCARCINCMA, NOS, INVASIVE

 1 

2 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY 20 50 50 

NATURAL DEATHS 15 

MORIBUND SACRIFICE 2 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 1 

TERMINAL SACRIFICE 18 31 29 

ANIMAL MISSING 2 

ANIMAL DELETED (WRONG SEX) 1 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

VEHICLE LOW DOSE HIGH DOSE
 

CONTROL
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 11 28 30
 

TOTAL PRIMARY TUMORS 11* 33 36
 

TOTAL ANIMALS WITH BENIGN TUMORS U 5 9
 

TOTAL BENIGN TUMORS 14 5 9
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 8 25 26
 

TOTAL MALIGNANT TUMORS 9 27 27
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 2 a 11
 

TOTAL SECONDARY TUMORS 3 5 22
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMOES
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PRIMARY OE METASTATIC 1 1
 

TOTAL UNCERTAIN TUHORS 1 1
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

# SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

55
 

APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN RATS ADMINISTERED TRIMETHYLP110SPHATE
 

BY GAVAGE
 

57
 

TABLE C1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

ADMINISTERED TRIMETHYLPHOSPHATE BY GAVAGE

 

VEHICLE LOW DOSE HIGH DOSE 

CONTROL 

ANIMAL S INITIALL Y IN STUDY 20 50 950 

ANIHALS NECROPSIEC 20 50 49 

ANIMAL S EXAMINE D HIS! [^PATHOLOGICALLY 20 50 49 

INTEGUMENTARY SYSTEM
 

*SUBCUT TISSUE (20) (50) (49)
 

DERMAL INCLUSION CYST 1 (2%)
 

INFLAMMATION, GRANULOMATOUS 1 (2X)
 

RESPIBATORY SYSTEM
 

f LUNG (19) (49) (46)
 

COLLAPSE 1 (2X)
 

CONGESTION, NOS 1 (2%) 1 (2X)
 

EDEMA, NOS 1 (2X)
 

HEMORRHAGE 1 (5X) 1 (2X) 1 (2X)
 

PNEUMONIA, CHRONIC MURINE 8 (42X) 21 (43X) 13 (28X)
 

HYPERPLASIA , ADENOMATOUS 1 (2X) 4 (9X)
 

METAPLASIA, NOS 1 (2X)
 

HEMATOPOIETIC SYSTEM
 

tBONE MARRO W (19) (42) (43)
 

HYPOPLASIA, NOS 1 (2%)
 

HfPERPLASIA, HEMATOPOIETIC 1 (2X) 3 (7X)
 

*SPLLEN (20) (47) (47)
 

CONGESTION, NOS 1 (2X) 1 (2X)
 

HhMORRHAGE 1 (2X)
 

FIBROSIS 2 (4X)
 

SCLEROSIS 1 (2%)
 

INFARCT, HEALED 1 <2X)
 

HEMOSIDEROSIS 1 (2X)
 

HYPERPLASIA, NOS 1 (2X)
 

HEMATOPOIESIS 1 (2X)
 

fMANDIBULAE L. NODE (20) (46) (46)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

3) 50 ANIMALS WERE INITIALLY IN THE STUDY, BUT ONE ANIMAL WAS FOUND TO EE A FEMALE
 

IN A BALE GROUP.
 

59
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE 

CONTROL 

LOW DOSE 

NECROSIS, NOS 

#MES£NTERIC I. NODE 

INFLAMMATION , CHRONIC 

HYPERPLASIA , PLASM A CELL 

(20) (46) 

1 (2X) 

1 (2X) 

CIRCULATOR Y SYSTEM 

#HEAHT/ATHIU M (20) (49) 

THROMBOSIS, NOS 3 (6%) 

ATHEROSCLEFOSI S 

*MYOCARDIUM (20) (49) 

INFLAMMATION , NOS 1 (2%) 

INFLAMMATION , CHRONIC FOCAL 

FIBROSIS 11 (55%) 29 (59X) 

FIBROSIS, DIFFUSE 2 («X) 

DIGESTIVE SYSTEM 

*SAL1VARY GLAN D (20) (43) 

HEMORRHAGI C CYST 

INFLAMMATION , ACUTE 1 (2X) 

GRANULATION , TISSUE 

ATROPHY , DIFFUSE 1 <2X ) 

*SALIVAR Y MUCOU S GLA N (20) (43) 

NUCLEAR-SHAP E ALTERATIO N 

#LIVER (20) (48) 

DEGENERATION , NOS 1 (5%) 

DEGENERATION , HYDROPIC 1 (2%) 

METAMORPHCSI S FATTY 2 (10X) 4 (8X) 

BASOPHILIC CYTO CHANG E 3 (15X) 2 (4%) 

CLEAR-CELL CHANG E 1 (5X) 

HYPERPLASIA , NODULA R 1 (2%) 

HYPERPLASIA , FOCAL 2 (1035) 3 (6%) 

#LIVEH/CF.NTRILOBULAR (20) (48) 

NECROSIS, NOS 

*BILE DUCT (20) (50) 

HY.PERPLASI.Aj. NOS. 7 J35%]_ 20 J40JL_ . 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE 

1 (2X) 

(46) 

(46) 

2 (4X) 

1 (2X) 

(46) 

1 (2%) 

27 (59%) 

1 (2X) 

(46) 

1 (255) 

1 (2% ) 

(46 ) 

1 (2% ) 

(48) 

1 (2% ) 

2 (4«) 

3 (6X) 

(48) 

1 (255) 

(49) 

21 _[43«1 

60
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE
 

(45)
 

3 (7%)
 

(45)
 

(47)
 

1 (2X)
 

(43)
 

(13)
 

1 (2X)
 

1 (2%)
 

11 (26%)
 

(48)
 

1 (2X)
 

38	 (79S)
 

2 (4X)
 

(48)
 

(48)
 

1 (2X)
 

(35)
 

1 (3X)
 

(38)
 

#PANCREAS
 

F1BROSIS, DIFFUSE
 

ATROPHY, FOCAL
 

#PANCREATIC ACINUS
 

ATROPHY, NOS
 

#STOBACH
 

ULCER, NOS
 

fLARGE INTESTINE
 

NEBATODIASIS
 

tCOLON
 

INFLAMMATION, ACUTE/CHRONIC
 

RUPTURE, INFLAMMATORY
 

PARASITISM
 

URINABY SYSTEM
 

#KIDNEY
 

GLOMERULONEPHRITIS, NOS
 

INPLAMBATICN, NOS
 

INFLAMMATICN, CHRONIC
 

FIBROSIS, DIFFUSE
 

NEPHROPATHY, TOXIC
 

tKIDNEY/GLOMERULUS
 

DEGENERATION, HYALINE
 

#KIDNEY/TUBULE
 

DILATATION, NOS
 

NECROSIS, NOS
 

#URINARY BLSDEER
 

DILATATION, NOS
 

ENDOCRINE SYSTEM
 

tPITUITARY
 

CYST, NOS
 

HUnORRHAGIC CYST ._... _
 

VEHICLE
 

CONTROL
 

(18)
 

1 (6X)
 

(18)
 

(15)
 

(12)
 

3 (25*)
 

(12)
 

(20)
 

13	 (65%)
 

1 (5X)
 

(20)
 

(20)
 

(11)
 

(16)
 

3 (19X)
 

LOW DOSE
 

(44)
 

3 (7%)
 

(44)
 

1 (2X)
 

(45)
 

(41)
 

(41)
 

6 (15X)
 

(49)
 

1 (2X)
 

1 (2X)
 

33	 (67X)
 

1 (2X)
 

(49)
 

1 (2X)
 

(49)
 

1 (2X)
 

1 (2X)
 

(36)
 

(44)
 

1 J241 ___.
 

I NUMBER OF ANIMALS WITH TISSOE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

61
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE
 

1 (3%)
 

(47)
 

1 (2X)
 

1 (2X)
 

1(2%)
 

(47)
 

(46)
 

1 (2X)
 

(49)
 

(49)
 

(46)
 

2 C»X)
 

(48)
 

(48)
 

1 (2X)
 

ANGIECTASIS
 

#ADRENAL


CYST, NOS
 

THROMBOSIS, NOS
 

GRANULOMA, NOS
 

LIPOIDOSIS
 

CALCIFICATION, NOS
 

#ADR£NAl CORTEX


HYPERPLASIA, NOS
 

#THYROID


HYPERPLASIA, C-CELL
 

HYPERPLASIA, FOLLICUL AR-CELL
 

REPRODUCTIVE SYSTEM
 

*MAMMARY GLAND


DILATATION/DUCTS


*SEHINAL VESICLE


CALCULUS, NOS
 

INFLAMMATION, ACUTE


INFLAMMATION, ACUTE FOCAL


tTESTIS


ATROPHY, NOS
 

NERVOUS SYSTEM
 

*BRAIN


MINERALIZATION


HEMORRHAGE
 

fBRAIN STEM


HEMORRHAGE
 

SPECIAL SENSE ORGANS
 

NONE
 

HUSCULOSKELETAL SYSTEM
 

VEHICLE
 

CONTROL
 

 (20)
 

 (20)
 

 (19)
 

 (20)
 

1 (595)
 

 (20)
 

1 (5X)
 

1 (5X)
 

 (16)
 

 (20)
 

1 (5%)
 

 (20)
 

LOW DOSE
 

(48)
 

1 (2%)
 

1 (2%)
 

(«8)
 

1 <2X)
 

(45)
 

5 (11*)


1 (2X)
 

(50)
 

(50)
 

1 12%)
 

(46)
 

1 (2%)
 

(50)
 

1 (2%)
 

(50)
 

1 (2X)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED BICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

62
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE

CONTROL 

 LOW DOSE HIGH DOSE 

BODY CAVITIES 

*ABDOMINAL CAVITY

NECROSIS, FAT

 (20)

 1 (5«)

 (50) (49) 

3 (6%) 

*BESENTERY

NECROSIS, FAT

 (20) (50)

 1 (2%) 

 (l»9) 

ALL OTHER SYSTEMS 

ADIPOSE TISSUE 

INFLAMMATION, GRANULOMATOOS

NECROSIS, FAT

 1 

1 

SPECIAL MORPHOLOGY SUMMARY 

NO LESION REPORTED

AUTO/NECROPSY/HISTO PERP

 1 1 1 

1 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

63
 
TABLE C2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

ADMINISTERED TRIMETHYLPHOSPHATE BY GAVA6E 

HIGH DOSE
 

950
 

49
 

48
 

(45)
 

1 (2X)
 

1 (2X)
 

3 (7X)
 

1 (2X)
 

16 (36X)
 

1 (2X)
 

4 (9X)
 

(37)
 

2 (5X)
 

(46)
 

3 (7X)
 

3 (7X)
 

(46)
 

(47)
 

ANIMALS
 

ANIMALS
 

ANIMALS
 

INITIALLY IN STUDY
 

NECROPSIED
 

EXAMINED HI STOPATHO LOGIC ALLY
 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEM
 

ILUNG
 

COLLAPSE
 

ATELECTASIS
 

CONGESTION, NOS
 

HEMORRHAGE
 

PNEUMONIA, CHRONIC MORINE
 

PERIVASCULAR CUFFING
 

FOAM-CELL
 

HYPERPLASIA, ADENOMATOUS
 

HEMATOPOIETIC SYSTEM
 

#BONE MARROW
 

HYPERPLASIA, HEMATOPOIETIC
 

fSPLEEN
 

CONGESTION, NOS
 

CONGESTION, CHRONIC
 

H£MOSIDEFOSIS
 

HEBATOPOIESIS
 

IMESENTERIC L. NODE
 

HASTOCYTOSIS
 

CIRCULATORY SYSTEM
 

*HEART
 

VEHICLE LOW DOSE
 

CONTROL
 

20
 

20
 

20
 

(20)
 

1 (5*)
 

10 (SOX)
 

1 (5X)
 

2 (10X)
 

(18)
 

(20)
 

(19)
 

(20)
 

50
 

50
 

50
 

(50)
 

5 (10X)
 

4 (8X)
 

19 (38X)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

(<t2)
 

(49)
 

1 (2X)
 

1 (2X)
 

3 (6X)
 

(46)
 

1 <2X)
 

(48)
 

* NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

64
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE LOW DOSE HIGH DOSE
 

CONTROL
 

tMYOCABDIUM (20) («8) (47)
 

INFLAMMATION, FOCAL 1 (2%)
 

FIBHOSIS 11 (55%) 23 («8%) 22 (47%)
 

DIGESTIVE SYSTEM
 

#SALIVARY GIAND (19) (46) (43)
 

DILATATION/DUCTS 1 (2%)
 

INFLAMMATION, CHRONIC 1 (2%)
 

ATROPHY, FOCAL 1 (2%)
 

fLIVhR (20) (48) («6)
 

GRANULOMA, NOS 1 (2%)
 

DEGENERATION, NOS 1 (2%)
 

BASOPHILIC CYTO CHANGE 9 (4555) 11 (23%) 11 (24%)
 

EOSINOPHILIC CYTO CHANGE 1 (2%)
 

CLEAR-CELL CHANGE 1 (2%)
 

CYTOLOGIC DEGENERATION 1 (2%) 1 (2%)
 

HYPERPLASIA, FOCAL 3 (15%) 7 (15%) 5 (11%)
 

*BILE DUCT (20) (50) (49)
 

HYPERPLASIA, NOS 4 (20%) 8 (16%) 8 (16%)
 

#PANCREAS (20) (48) (46)
 

FIBROSIS, DIFFUSE 2 (4%)
 

ATROPHY, NOS 1 (2%) 1 (2%)
 

ATROPHY, FCCAL 2 (10%) 2 (4%) 2 (4%)
 

fPANCEEATIC ACINUS (20) (48) (46)
 

ATROPHY, NOS 1 (2%)
 

ATROPHY, DIFFUSE 1 (2%)
 

#ILEUM (17) (44) (43)
 

HYPERPLASIA, LYMPHOID 1 (2%)
 

#LARGE INTESTINE (16) (44) (43)
 

NEMATODIASIS 4 (9%)
 

f COLON (16) (44) (43)
 

PARASITISM 2 (13%) 6 (14%)
 

URINAR Y SYSTEM 

fKIDNE Y (20) (50) (48)

 

HEMORRHAGIC_CYS T 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* «uMBER OF ANIMALS NECROPSIED
 

65
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE
 

28 (58X)
 

(48)
 

1 (2X)
 

(31)
 

(41)
 

5 (12%)
 

(48)
 

3 (6X)
 

2 (4%)
 

(48)
 

1 (2X)
 

(43)
 

1 (2%)
 

(49)
 

1 (2X)
 

1 <2X)
 

(44)
 

1 (2%)
 

(44)
 

1 (2%)
 

2 (5%)
 

1 J2XJ. .
 

PYELONEPHRITIS, NOS
 

INFLAMHATICN, CHRONIC
 

NEPHROSIS, CHOLEMIC
 

#KIDNEY/TUBULE
 

NECROSIS, NOS
 

NECROSIS, FOCAL
 

#URINARY BLAEDER
 

INFLAMMATION, NOS
 

METAPLASIA, SQUAMOUS
 

ENDOCBINE SYSTEM
 

tPITUITARY
 

CYST, NOS
 

HEMORRHAGIC CYST
 

ANGIECTASIS
 

#ADRtNAL
 

LIPOIDOSIS
 

ANGIECTASIS
 

•ADRENAL	 CORTEX
 

HYPERPLASIA, FOCAL
 

#THYROID
 

HYPERPLASIA, C-CELL
 

REPRODUCTIVE SYSTEM
 

*MAHMARY GLAND
 

DILATATICN/DUCTS
 

HYPERPLASIA, NOS
 

f UTERUS
 

HEMORRHAGE
 

HYPERPLASIA, STHOMAL
 

tUTERUS/ENDOMETRIUM
 

INFLAMMATION, ACUTE
 

HYPERPLASIA, NOS
 

HY,PERPLASIA,._CYSTIC_
 

VEHICLE
 

CONTROL
 

14 (10%)
 

(20)
 

1 (558)
 

(17)
 

1 (6X)
 

1 (6%)
 

(20)
 

1 (5X)
 

1 (5%)
 

(19)
 

(19)
 

(19)
 

(20)
 

(20)
 

1 (5%)
 

(20)
 

2 (10%)
 

LOW DOSE
 

1 (2X)
 

28 (56X)
 

1 (2*)
 

(50)
 

1 (2%)
 

(32)
 

(i»8)
 

4 (8%)
 

3 (6X)
 

(1*8)
 

1 (2%)
 

(U8)
 

(47)
 

2 (<*«)
 

(50)
 

(i»5)
 

1 (2%)
 

(45)
 

2 (UX)
 

# NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

66
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE

CONTROL 

 LOW DOSE HIGH DOSE 

*OVAnY

FOLLICOLAR CYST, NOS

 (20)

 1 (5X)

 (U5)

 1 (2%)

 (l»3) 

1 (2%) 

NERVOUS SYSTEM 

#BRAI N

MINERALIZATIO N

 (20) (50) (U8 ) 

1 (2%) 

SPECIAL

NON E 

 SENSE ORGAN S 

HUSCUiOSKELETA L SYSTEM 

NONjc J 

BODY CAVITIE S 

*ABDOMINA L CAVIT Y

NECROSIS, FAT

 (20 ) (50) (<*9) 

1 (2%) 

A LL OTHER SYSTEM S 

NONE 

SPECIAL MORPHOIOG Y SUMMAR Y 

NO LESION EEPOBTED

AUTO/NECROPSY/NO HISTO

t NUMBER OF ANIMAL S WIT H TISSUE

* NUHflER OF ANIMALS NECROPSIED 

 EXAMINE D MICROSCOPICALLY 

1 1 

1 

67 

APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN MICE ADMINISTERED TRIMETHYLPHOSPHATE
 

BY GAVACE
 

69
 

TABLE D1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

ADMINISTERED TRIMETHYLPHOSPHATE BY GAVAGE 

VEHICLE LOW DOSE
 

CONTROL
 

ANIMALS INITIALLY IN STUDY 20 50
 

'ANIMALS NECEOPSIBD 20 50
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 20 50
 

INTEGUMENTARY SYSTEM
 

*SKIN (20) (50)
 

HEMORRHAGIC CYST
 

INFLAMMATION, CHRONIC 1 (2«)
 

RESPIfiATORY SYSTEM
 

tLUNG (20) (50)
 

INFLAMMATION, FOCAL 1 (2%)
 

INFLAMMATION, ACUTE
 

PNEUMONIA, CHRONIC MURINE
 

INFLAMMATION, FOCAL GRANULOHATOU 1 (5X) 1 (2X)
 

PERIVASCULAR CUFFING
 

CRYSTALS, NOS 1 (5X)
 

HYPERPLASIA, ADENOMATOUS 3 (6X)
 

METAPLASIA, OSSEOUS
 

HISTIOCYTOSIS
 

tLUNG/ALVEOLI (20) (50)
 

HISTIOCYTOSIS 1 (5«)
 

HEMATOPOIETIC SYSTEM
 

fSPLEEN (18) (45)
 

HYPERPLASIA, LYMPHOID 1 (6%)
 

HEMATOPOIESIS 1 (6X) 1 (2X)
 

#LYBPH NODE (18) (45)
 

HEMORRHAGE 1 (2%)
 

IHANDIBULAB L. NODE (18) (45)
 _EIiASHAC.IlfisJ£_ _ _ _
 

* NOHBEB OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMAL S NECROPSIED
 

HIGH DOSE
 

49
 

49
 

49
 

(49)
 

1 (2X)
 

(49)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

1 (2%)
 

2 (4X)
 

(49)
 

(44)
 

(41)
 

(41)
 

1 -12X1
 

71
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE LOW DOSE 

CONTROL 

fMESENTERIC L. NODE (18) (15) 

HEMORRHAG E 2 (IX) 

HYPERPLASIA , LYWPHOI D 1 (2X) 

HEMATOPOIESIS 1 (6X) 1 (2X) 

CIRCULATOR Y SYSTEM 

*MYOCARDIU M (19) (48)

 

INFLAMMATION , CHRONIC FOCAL 

FIBHOSIS 1 (5X)

 

FIBROSIS, IOCAL 

CALCIFICATION , FOCAL 

*ARTER Y (20) (50) 

INFLAMMATION , CHRONIC 

MEDIA L CALCIFICATIO N 1 (2X) 

DIGESTIV E SYSTE M 

#LIVE R (20) (48) 

CYST, NOS 1 (5X) 

INFLAMMATION , CHRONIC FOCAL 3 (6X) 

DEGENERATION , HYDROPIC 

NECROSIS , FOCAL 1 (5X) 

Cl'TOPLASMIC VACUOLIZATION 1 (2X) 

HYPERPLASIA , FOCAL 1 (5X) 

ANGIECTASIS 1 (5X) 

*BIL£ DUCT (20) (50) 

INFLAMMATION , CHRONIC 1 (2X) 

IPANCREA S (20) (46) 

DILATATION/DUCTS 1 (2X) 

INFLAMMATION , CHRONIC DIFFUSE 1 (2X) 

#PANCREATI C ACINU S (20) (46) 

ATROPHY, NOS 2 (4X) 

ATROPHY , FCCAL 1 (2X) 

tSTOMAC H (18) (50)

 

INFLAMMATION , ACUTE 

*PEYERS PATC H (20) (49) HYPPRPTASTANO S 1 ( 2%) 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE 

(11)

2 (5X) 

(i»7) 

1 (2X) 

2 (4X) 

1 (2X) 

(19) 

1 (2X) 

(49) 

1 <2X) 

1 (2X) 

1 (2X) 

1 (2X) 

(49) 

(12) 

(42) 

(16) 

1 (2X) 

(15) 

72
 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE LOW DOSE HIGH DOSE 

CONTROL 

ICOLON (20) (U7) 

NEMATODIASIS 1 (2%) 

PARASITISM 3 (15%) 12 (25%) 21 (USX) 

UillliBT SY3TBH 

IKIDMEY (20) (49) (49)
 

IHFLABMATICN. CHRONIC 1 (5%)
 

NiPHROPATHY, TOXIC 1 (2X)
 

LYMPHOCYTOSIS 1 (2%)
 

#KIDNEY/GLOMERULUS (20) (49) (a 9)
 

INFLAMMATION, CHRONIC 1 (2%)
 

#KIDNEY/TUBOLE (20) (U9) (19)
 

ATROPHY, FOCAL 1 (2«)
 

f
 

ENDOCRINE SYSTEM
 

*THYBOID (9) (35) (39)
 

POLLICOLAH CYST, SOS 1 (3X)
 

HYPBRPLASIA, ADSNOHATOUS 1 (3%)
 

REPRODUCTIVB SYSTBH
 

NONE
 

NERVOUS SYSTEH
 

• BRAI N (20) (49) (48) 

CALCIFICATION, FOCAL 9 (45%) 15 (31%) 15 (31%) 

SPECIAL SENSE C8GANS 

NON E 

MUSCULOSKELETAL SYSTEM 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NBCROPSIED
 

73 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE LOW DOSE HIGH DOSE 

CONTROL 

BODY CAVITIES 

NONE 

ALL OIHER SYSTEMS 

ADIPOSE TISSUE 

INFLAMMATION . CHRONIC 

SPECIAL MORPHOLOG Y SUMMAR Y 

NO LESION REPORTED 

t NUMBER OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

74 
TABLE D2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

ADMINISTERED TRIMETHYLPHOSPHATE BY GAVAGE 

ANIMALS INITIAILY IN STUDY
 

ANIMALS MISSING
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEM
 

&LUNG/BRONCHUS
 

CRYSTALS, NOS
 

#LUNG
 

THROMBOSIS, NOS
 

PNEUMONIA, CHRONIC MURINE
 

INFLAMMATION, FOCAL GRANULOMATOU
 

INFAPCT, NCS
 

HYPERPLASIA, ADENOMATOUS
 

HISTIOCYTCSIS
 

LEUKEMOID REACTION
 

HEMATOPOIETIC SYSTEM
 

#SPL£EN
 

HYPERPLASIA, RETICULUM CELL
 

HYPERPLASIA, LYMPHOID
 

HiMATOPOIESIS
 

#MESENTERIC L. NODE
 

HEMORRHAGE
 

INFLAMMATION, GRANU LOHATOU S
 

HYPERPLASIA, RETICULUM CELL
 

CIRCULATORY SYSTEM
 

*BLOOD VESSEL


INFLAMMATICNi_CH£ONIC


VEHICLE LOW DOSE HIGH DOSE CONTROL 

20 50 950 

2 

20 50 1*7 

20 50 1*7 

(20) (1*8) (<»5) 

1 (5%) 1 (2%) 

(20) (1*8) (1,5) 

1 (2%) 

6 (30%) 13 (21%) 6 (13%) 

1 (2%) 1 

1 

(2%) 

(2%) 

2 (14%) 

2 (U %) 

2 (U%) 

(17) (48) (*»!*) 

1 (6%) 

2 C*X) 

1 (2«) 

5 (10%) 

1 (2%) 

1 (2X) 

3 (7%) 

(17) (46) («3) 

1 (2%) 

1 (2*) 

1 (2*) 

 (20) (50) (1*7) 

 -!_i2%J 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

3 50 ANIMALS WERE INITIALLY IN THE STUDY, BUT ONE ANIMAL WAS FOUND TO EE A MALE
 

IN A FEMALE GROUP.
 

75
 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE LOW DOSE 

CONTROL 

*PULBONARY AETEE Y (20) (50) 

THROHBOSIS, NOS 

DIGESTIVE SYSTE M 

#LIVER (20) (50) 

INFLAMMATION, NEC80TIZING 

INFLABMATION , CHRONIC FOCAL 1 (5*) 1 (2X) 

INFLAMMATION , GRANULOMATOU S 1 (5*) 

INFLAMMATION , FOCAL GRANULOMATOO 1 (2X) 

PiRIVASCULAR CUFFING 

DEGENERATION , HYDROPIC 

NECROSIS, FOCAL 1 (5%) 

1 (2X) 

1 (2%) 

METAMOBPHOSIS FATTY 1 (5%) 

CYTOPLASMIC VACOOLIZ ATION 

HYPERPLASIA, FOCAL 

HEMATOPOIESIS 1 (2X) 

tLIVER/CENTRILOBULAR (20) (50) 

NECROSIS, NOS 1 (2X) 

#LIV£R/PERIPOBTAL (20) (50) 

INFLAMMATION , CHRONIC 1 (5X) 

*BILE DUCT (20) (50) 

DILATATION, NOS 1 (2X) 

INFLAMMATION , CHRONIC 1 (5%) 2 (<*X) 

•PANCREAS (20) (<*8) 

INFLAMMATION, CHROMIC FOCAL 

tPANCREATIC ACINUS (20) (48) 

ATROPHY, NOS 2 (<•*) 

ATROPHY, FOCAL 

tSTOMACH (17) (46) 

INFLAMMATION , ACUTE FOCAL 1 (2X) 

MASTOCYTOSIS 1 (2X) 

tSMALL INTESTINE (19) (48) 

INFLAMMATION , GRANULOMATOUS 1 (5X) 

AMYLOIDOSIS 

*PBY£RS PATCH (19) (48) 

_ HIPIHPLASlfiA_BQ§_ _ - i ­, 1 12SL 

t NUMBER OF ANIMALS HITB TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NBCROPSIED
 

HIGH DOSE 

(47) 

2 (4») 

(44) 

1 (2X) 

1 (2X) 

1 (2X) 

1 (2X) 

1 (2X) 

1 (2X) 

(44) 

(44) 

(47) 

(t2) 

1 (2X) 

(12) 

2 (5X) 

1 (2X) 

(47) 

(43) 

1 (2») 

(<«3)

JLiiSi 

76
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

fCOLON
 

PARASITISM
 

URINABY SYSTEM
 

#KIDNEY
 

HYDRONEPHROSIS
 

INFLAMMATION, ACUTE FOCAL
 

INFLAMMATION, CHRONIC
 

PERIVASCULAR CUFFING
 

NEPHROPATHY, TOXIC
 

#KIDNEY/TUBULE
 

CALCIFICATION, NOS
 

HtMOSIDEEOSIS
 

fURINARY BLADDER
 

HEMORRHAGIC CYST
 

ENDOCBINE SYSTEM
 

#ADRENAL
 

INFLAMMATION, CHRONIC FOCAL
 

REPRODUCTIVE SYSTEM
 

fUTEKUS
 

HYDROMETRA
 

INFLAMMATION, SUPPURATIVE
 

PYOMETRA
 

ABSCESS, NOS
 

#UTEKUS/ENDOMETRIUM
 

CYST, NOS
 

MULTIPLE CYSTS
 

HYPERPLASIA, CYSTIC
 

IUTEBUS/M YOMETRIUM
 

INFLAMMATION, CHRONIC FOCAL
 

*OVABY
 

_£ISTr_NOS _
 

VEHICLE
 

CONTROL
 

(19)
 

2 (11*)
 

(19)
 

1 (5X)
 

(19)
 

(13)
 

(17)
 

(16)
 

1 (6X) .
 

(16)
 

(16)
 

(10)
 

LOW DOSE
 

(49)
 

11 (22%)
 

(50)
 

3 <6X)
 

1 (2X)
 

1 (2X)
 

(50)
 

1 (2X)
 

(27)
 

1 (4%)
 

(46)
 

1 (2X)
 

(40)
 

1 (3%)
 

1 (3X)
 

1 OX)
 

(40)
 

4 (10%)
 

2 (5%)
 

(40)
 

1 (3%)
 

(26)
 

4 -Q5X)
 

HIGH DOSE
 

(17)


9 (19X)
 

(44)
 

4 (9%)
 

1 (2X)
 

2 (5%)
 

(44)
 

1 (2%)
 

(25)
 

(39)
 

(37)
 

1 (3X)
 

(37)
 

1 (3%)
 

(37)
 

(28)
 

_3 J1JS1-.
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECROPSIED
 

77
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

NERVOUS SYSTEM 

*BRAI N

HEMORRHAG E

CALCIFICATION, FOCAL

SPECIAL SENSE ORGAN S 

NONE 

HUSCULOSKELETAI SYSTEM 

*BONE


FIBROUS OS1EODYSTROPHY


OSTEOSCLEROSIS


*SKELETAL MOSCLE


INFLAHMATION, NOS


INFLAMMATION, CHRONIC FOCAL


BODY CAVITIES
 

*PERITONEUM


INFLAMMATION, CHRONIC FOCAL


*MESENTERY

NilCROSIS, JAT

ALL OTHER SYSTEMS 

NONE 

SPECIAL MORPHOLOG Y SUMMAR Y 

N O LESION REPORTED

ANIMA L HISSING/NO NECROPSY

* NUMBER OF ANIMALS KITH TISSUE

* NUMBER OF ANIHALS NECROPSIED 

VEHICLE LOW DOSE HIGH DOSE 

CONTROL 

 (19) (49) (46) 

1 (2%) 

6 (32%) 8 (16%) 8 (17%) 

 (20) (50) (47) 

 11 (55%) 19 (38%) 11 (23%) 

1 (5%) 1 (2%) 1 (2%) 

 (20) (50) (47) 

1 (2%) 

2 (10%) 

 (20) (50) (47) 

1 (2%) 

 (20) (50) (47) 

1 (2%) 

1 1 3 

2 

 EXAMINE D MICROSCOPICALLY 

78 
APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN
 

RATS ADMINISTERED TRIMETHYLPHOSPHATE
 

BY GAVAGE
 

79
 

Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered Trimethylphosphate by Gavagea
 

Topography: Morphology

Vehicle 

 Control 

Subcutaneous Tissue: 

Fibromab 0/20 (0) 

P Values0 » d P ­ 0.006 

Relative Risk (Vehicle Control)f 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

oo 

Lung: Alveolar /Bronchiolar 

Adenoma or Carcinomab 0/19 (0) 

P Valuesc » d N.S 

Relative Risk (Vehicle Control)f 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor — 

Low
 

Dose
 

2/50 (4)
 

N.S.
 

Infinite
 

0.123
 

Infinite
 

67
 

2/49 (4)
 

N.S.
 

Infinite
 

0.119
 

Infinite
 

105
 

High
 

Dose
 

9/49 (18)
 

P = 0.036
 

Infinite
 

1.119
 

Infinite
 

91
 

5/46 (11)
 

N.S.
 

Infinite
 

0.545
 

Infinite
 

84
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats 

Administered Trimethylphosphate by Gavage3 

(continued) 

Vehicle Low
 

Topography: Morphology Control Dose
 

Hematopoietic System:
 

Leukemia or Lymphomab

 8/20 (40) 20/50 (40)
 

P Valuesc»d N.S. N.S.
 

Relative Risk (Vehicle Control)f 1.000
 

Lower Limit 0.529
 

Upper Limit 2.242
 

Weeks to First Observed Tumor 81 79
 oo 

t-0 

Pituitary: Chromophobe
 

Adenoraab 4/16 (25) 13/44 (30)
 

P Valuesc'd N.S. N.S.
 

Relative Risk (Vehicle Control)f 1.182
 

Lower Limit 0.450
 

Upper Limit 4.430
 

Weeks to First Observed Tumor 83 79
 

High
 

Dose
 

25/49 (51)
 

N.S.
 

1.276
 

0.703
 

2.746
 

82
 

8/38 (21)
 

N.S.
 

0.842
 

0.276
 

3.406
 

92
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats 

Administered Trimethylphosphate by Gavagea 

(continued) 

Vehicle Low
 

Topography: Morphology Control Dose
 

Adrenal: Pheochromocytomab 1/20 (5) 4/48 (8)
 

P Valuesc»d N.S. N.S.
 

Relative Risk (Vehicle Control)f 1.667
 

Lower Limit 0.182
 

Upper Limit 80.315
 

Weeks to First Observed Tumor 105 93
 

oo Thyroid: C-cell Adenoma
 03

 or Carcinomab 1/19 (5) 3/45 (7)
 

P Valuesc»d N.S. N.S.
 

Relative Risk (Vehicle Control)f 1.267
 

Lower Limit 0.112
 

Upper Limit 64.997
 

Weeks to First Observed Tumor 105 67
 

High
 

Dose
 

7/47 (15)
 

N.S.
 

2.979
 

0.429
 

131.059
 

97
 

2/46 (4)
 

N.S.
 

0.826
 

0.047
 

47.694
 

97
 
00 

Table El. Analyses of the Incidence of Primary Tumors in Male Rats 

Administered Trimethylphosphate by Gavage3 

(continued) 

Vehicle Low High
 

Topography: Morphology Control Dose Dose
 

Testis: Interstitial-cell
 

Tumorb 11/16 (69) 33/46 (72) 25/46 (54)
 

P Values0 »d N.S. N.S. N.S.
 

Relative Risk (Vehicle Control)f 1.043 0.791
 

Lower Limit 0.742 0.544
 

Upper Limit 1.722 1.393
 

Weeks to First Observed Tumor 97 74 78
 

a

Dosed groups received 50 or 100 mg/kg.
 

"Number of tumor-bearing animals/number of animals examined at site (percent).
 

c

Beneath the incidence of tumors in the vehicle-control group is the probability level for the
 

Cochran- Armitage test when P < 0.05, otherwise, not significant (N.S.) is indicated. Beneath the
 

incidence of tumors in each dosed group is the probability level for the Fisher exact test for
 

the comparison of that dosed group with the vehicle-control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

"A negative trend (N) indicates a lower incidence in a dosed group than in the control group.
 

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

95 percent confidence interval of the relative risk between each dosed group and the
 

vehicle-control group.
 
__ 

__ 

Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Trimethylphosphate by Gavage3
 

Vehicle Low
 

Topography: Morphology Control Dose
 

Lung: Alveolar/Bronchiolar
 

Adenoma or Carcinomab 0/20 (0) 0/50 (0)
 

P Valuesc»d N.S. N.S.
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 —
 

—
 

Weeks to First Observed Tumor —
 

GO
 

Ui Hematopoietic System:
 

Lymphona or Leukemiab 3/20 (15) 14/50 (28)
 

P Valuesc»d N.S. N.S.
 

Relative Risk (Vehicle Control)f 1.867
 

Lower Limit 0.609
 

Upper Limit 9.359
 

Weeks to First Observed Tumor 98 86
 

High
 

Dose
 

3/45 (7)
 

N.S.
 

Infinite
 

0.278
 

Infinite
 

105
 

12/49 (24)
 

N.S.
 

1.633
 

0.513
 

8.342
 

59 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Trimethylphosphate by Gavagea
 

(continued)
 

Vehicle Low High
 

Topography: Morphology Control Dose Dose
 

Pituitary: Chromophobe Adenoma^ 9/20 (45) 21/48 (44) 18/41 (44)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Risk (Vehicle Control )f 0.972 0.976
 

Lower Limit 0.544 0.532
 

Upper Limit 2.025 2.049
 

Weeks to First Observed Tumor 98 86 93
 

oo Thyroid: C-cell Carcinomab

 2/19 (11) 0/47 (0) 2/43 (5)
 

o\
 

P Values0 »d N.S. N.S. N.S.
 

Departure from Linear Trend6 P - 0.044
 

Relative Risk (Vehicle Control)f 0.000 0.442
 

Lower Limit 0.000 0.035
 

Upper Limit 1.357 5.796
 

Weeks to First Observed Tumor 97 105
 —
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Trimethylphosphate by Gavage3
 

(continued)
 

Topography: Morphology


Thyroid: C-cell Adenoma
 

or Carcinomab


P Valuesc»d


Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


oo
 

Mammary Gland:
 

Fibroadenomab


P Valuesc»d


Relative Risk (Vehicle Control)*
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Vehicle
 

 Control
 

 2/19 (11)
 

 N.S.
 

 97
 

 2/20 (10)
 

 N.S.
 

 97
 

Low
 

Dose
 

6/47 (13)
 

N.S.
 

1.213
 

0.247
 

11.660
 

99
 

3/50 (6)
 

N.S.
 

0.600
 

0.076
 

6.860
 

101
 

High
 

Dose
 

5/43 (12)
 

N.S.
 

1.105
 

0.205
 

10.982
 

105
 

5/49 (10)
 

N.S.
 

1.020
 

0.188
 

10.204
 

96
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats 

Administered Trimethylphosphate by Gavagea 

(continued) 

Vehicle Low High
 

Topography: Morphology Control Dose Dose
 

Uterus: Endometrial
 

Stroraal Polypb 2/20 (10) 1/45 (2) 0/44 (0)
 

P Valuesc»d P = 0.043 (N) N.S. N.S.
 

Relative Risk (Vehicle Control)f 0.222 0.000
 

Lower Limit 0.004 0.000
 

Upper Limit 4.077 1.524
 

Weeks to First Observed Tumor 105 105
 

oo —
 oo a

Dosed groups received 50 or 100 mg/kg.
 

^Number of tumor-bearing animals/number of animals examined at site (percent).
 

c

Beneath the incidence of tumors in the vehicle-control group is the probability level for the
 

Cochran-Armitage test when P < 0.05, otherwise, not significant (N.S.) is indicated. Beneath the
 

incidence of tumors in each dosed group is the probability level for the Fisher exact test for
 

the comparison of that dosed group with the vehicle-control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

"A negative trend (N) indicates a lower incidence in a dosed group than in the control group.
 

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

*The 95 percent confidence interval of the relative risk between each dosed group and the
 

vehicle-control group.
 
APPENDIX F
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN
 

MICE ADMINISTERED TRIMETHYLPHOSPHATE
 

BY GAVAGE
 

89
 

Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered Triraethylphosphate by Gavagea
 

Topography: Morphology


Lung: Alveolar/Bronchiolar
 

Carcinomab


P Values0 »d


Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Lung: Alveolar/Bronchiolar
 

Adenoma or Carcinoma

b


P Valuesc»d


Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Vehicle
 

 Control
 

 2/20 (10)
 

 N.S.
 

 103
 

 3/20 (15)
 

 N.S.
 

 103
 

Low
 

Dose
 

8/50 (16)
 

N.S.
 

1.600
 

0.364
 

14.699
 

103
 

11/50 (22)
 

N.S.
 

1.467
 

0.450
 

7.594
 

103
 

High
 

Dose
 

3/49 (6)
 

N.S.
 

0.612
 

0.078
 

6.996
 

81
 

9/49 (18)
 

N.S.
 

1.224
 

0.354
 

6.533
 

81
 
Table Fl.


(continued)
 

Topography: Morphology


Hematopoietic System:
 

Lymphoma or Leukemiab


P Valuesc»d


Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Liver: Hepatocellular
 

Carcinoma6


P Valuesc»d


Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


 Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered Trimethylphosphate by Gavage3
 

Vehicle Low High 

 Control Dose Dose 

 3/20 (15) 5/50 (10) 9/49 (18) 

 N.S. N.S. N.S. 

0.667 1.224 

0.147 0.354 

4.014 6.533 

 97 82 90 

 4/20 (20) 9/48 (19) 8/49 (16) 

 N.S. N.S. N.S. 

0.938 0.816 

0.307 0.255 

3.804 3.392 

 89 103 98 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered Trimethylphosphate by Gavage3
 

(continued)
 

Topography: Morphology

Vehicle 

 Control 

Low 

Dose 

High 

Dose 

Liver: Hepatocellular 

Adenoma or Carcinoma"-1 4/20 (20) 10/48 (21) 8/49 (16) 

P Valuesc'd N.S. N.S. N.S. 

Relative Risk (Vehicle Control)f 

Lower Limit 

Upper Limit 

1.402 

0.353 

4.148 

0.816 

0.255 

3.392 

Weeks to First Observed Tumor 89 103 98 

a

Dosed groups received 250 or 500 mg/kg. 

"Number of tumor-bearing animals /number of animals examined at site (percent).
 

c

Beneath the incidence of tumors in the vehicle-control group is the probability level for the
 

Cochran-Armitage test when P < 0.05, otherwise, not significant (N.S.) is indicated. Beneath the
 

incidence of tumors in each dosed group is the probability level for the Fisher exact test for
 

the comparison of that dosed group with the vehicle-control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

°*A negative trend (N) indicates a lower incidence in a dosed group than in the control group.
 

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

95 percent confidence interval of the relative risk between each dosed group and the
 

vehicle-control group.
 
__ 

Table F2. Analyses of the Incidence of Primary Tumors in Female Mice 

Administered Trimethylphosphate by Gavagea 

Vehicle Low High
 

Topography: Morphology Control Dose Dose
 

Lung: Alveolar/Bronchiolar
 

Adenoma or Carcinoma'' 3/20 (15) 0/48 (0) 6/45 (13)
 

P Valuesc»d N.S. P - 0.023 (N) N.S.
 

Departure from Linear Trend6 P = 0.008
 

Relative Risk (Vehicle Control)^ 0.000 0.889
 

Lower Limit 0.000 0.218
 

Upper Limit 0.686 5.104
 

Weeks to First Observed Tumor 99 80
 

Hematopoietic System:
 

Lymphoma or Leukemiab 5/20 (25) 14/50 (28) 11/47 (23)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Risk (Vehicle Control)f 1.120 0.936
 

Lower Limit 0.457 0.358
 

Upper Limit 3.556 3.080
 

Weeks to First Observed Tumor 99 36 59
 
__ 

Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered Trimethylphosphate by Gavage3
 

(continued)
 

Vehicle Low
 

Topography: Morphology Control Dose
 

Liver: Hepatocellular
 

Adenoma or Carcinomab 2/20 (10) 4/50 (8)
 

P Valuesc»d N.S. N.S.
 

Relative Risk (Vehicle Control)f 0.800
 

Lower Limit 0.128
 

Upper Limit 8.436
 

Weeks to First Observed Tumor 103 86
 

vo
 

Ul Uterus /Endoraetrium:
 

Adenocarcinoma, NOSb 0/16 (0) 7/40 (18)
 

P Valuesc»d P = 0.003 N.S.
 

Relative Risk (Vehicle Control)f Infinite
 

Lower Limit 0.825
 

Upper Limit Infinite
 

Weeks to First Observed Tumor — 94
 

High
 

Dose
 

0/44 (0)
 

N.S.
 

0.000
 

0.000
 

1.524
 

13/37 (35)
 

P = 0.004
 

Infinite
 

1.850
 

Infinite
 

83
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice 

Administered Trimethylphosphate by Gavagea 

(continued) 

Vehicle Low High
 

Topography: Morphology Control Dose Dose
 

Uterus: Endometrial Stromal
 

Polypb 0/16 (0) 2/40 (5) 1/37 (3)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Risk (Vehicle Control)f Infinite Infinite
 

Lower Limit 0.125 0.024
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor — 93 103
 

O\ a

Dosed groups received 250 or 500 mg/kg. 

c

Beneath the incidence of tumors in the vehicle-control group is the probability level for the
 

Cochran-Armitage test when P < 0.05, otherwise, not significant (N.S.) is indicated. Beneath the
 

incidence of tumors in each dosed group is the probability level for the Fisher exact test for
 

the comparison of that dosed group with the vehicle-control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

°*A negative trend (N) indicates a lower incidence in a dosed group than in the control group.
 

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

•'-The 95 percent confidence interval of the relative risk between each dosed group and the
 

vehicle-control group.
 
Review of the Bioassay of Trimethylphosphate* for Carcinogenicli
 

by the Data Evaluation/Risk Assessment Subgroup of the
 

Clearinghouse on Environmental Carcinogens
 

March 6, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance with DHEW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemicai
 

carcinogens in the environment to which humans may be
 

exposed. The members of the Clearinghouse have been drawn
 

from academia, industry, organized labor, public interest


groups, State health officials, and quasi-public health and
 

research organizations. Members have been selected on the
 

basis of their experience in carcinogenesis or related field?
 

and, collectively, provide expertise in chemistry, biochemist,
 biostatistics, toxicology, pathology, and epidemiology.
 Representatives of various Governmental agencies participate
 

as ad hoc members. The Data Evaluation/Risk Assessment Subgr.,
 of the Clearinghouse is charged with the responsibility of
 

providing a peer review of reports prepared on NCI-sponsored
 

bioassays of chemicals studied for carcinogenicity. It is
 

this context that the below critique is given on the bioassa,
 of Trimethylphosphate for carcinogenicity.
 

The primary reviewer noted that Trimethylphosphate is
 

used as a gas additive and in plastics as a flame retardant,.
 After a brief description of the conditions of test, he sa_u­ that the compound was associated with a higher incidence o:
 

fibromas and other tumors randomly distributed in this treat­ ment group. In the treated female mice, Trimethylphosphate
 

induqed a statistically significant incidence of adenocarcir j


of the uterus/endometrium. Testicular interstitial-cell tur
 

were found in two low dose treated mice. The primary review-
 opined that the effects on the reproductive organs may be
 

associated with the sterilent property of the chemical. 3n
 

summary, he said that he agreed with the conclusion in the
 

report that Trimethylphosphate was carcinogenic in at least
 

the female mice.
 

The secondary reviewer said that there was a clear
 

dose-response in the incidence of uterine/endometrium adene­ carcinomas in treated female mice. After some discussion
 

regarding the language of the conclusion in the report, it
 

97
 
was recommended that it be reworded to reflect the uncertainty
 

regarding the carcinogenicity of Trimethylphosphate in the
 

other treatment groups.
 

It was moved that the report be accepted as modified.
 

The motion was seconded and approved unanimously.
 

Members present were
 

Gerald N. Wogan (Chairman), Massachusetts Institute of
 

Technology
 

Arnold Brown, Mayo Clinic
 

Lawrence Garfinkel, American Cancer Society
 

E. Cuyler Hammond, American Cancer Society
 

Joseph Highland, Environmental Defense Fund
 

Henry Pitot, University of Wisconsin Medical Center
 

George Roush, Jr., Monsanto Company
 

Sheldon Samuels, Industrial Union Department, AFL-CIO
 

Michael Shimkin, University of California at San Diego
 

John Weisburger, American Health Foundation
 

Sidney Wolfe, Health Research Group
 

Subsequent to this review, changes may have been made
 

in the bioassay report either as a result of the review
 

or other reasons. Thus, certain comments and criticisms
 

reflected in the review may no longer be appropriate.
 

1J-U.S. GOVERNMEN T PRINTIN G OFFICE : 1978-260-899/3160
 

98
 

DHEW Publication No. (NIH) 78-1331