T National Cancer Institute 

-H 

I CARCINOGENESIS 

Technical Report Series 

= NO. 159 

£ 1979 

I 

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BIOASSAY OF 

OFFICIAL NTP
 

FILE COPY
 

DO NOT REMOVE FROM THIS OFFICE
 

PHTHALIC ANHYDRIDE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 85-44-9 

NCI-CG-TR-159 

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

Public Health Service 

National Institutes of Health 

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BIOASSAY OF
 

PHTHALIC ANHYDRIDE
 

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) 79-1715
 
ii

 
BIOASSAY OF
 

PHTHALIC ANHYDRIDE
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

FOREWORD; This report presents the results of the bioassay of
 

phthalic anhydride 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 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
 

chemicals found to be carcinogenic in animals requires a wider
 

analysis.
 

CONTRIBUTORS: This bioassay of phthalic anhydride was conducted
 

by the NCI Frederick Cancer Research Center (FCRC) (1),
 

Frederick, Maryland, operated for NCI (2) by Litton Bionetics,
 

Inc.
 

The manager of the bioassay at FCRC was Dr. B. Ulland, the
 

toxicologist was Dr. E. Gordon, and Drs. R. Cardy and D. Creasia
 

compiled the data. Ms. S. Toms was responsible for management of
 

data, Mr. D. Cameron for management of histopathology, Mr. L.
 

Callahan for management of the computer branch, and Mr. R. Cypher
 

for the management of the facilities. Mr. A. Butler performed
 

the computer services. The histopathology of early deaths was
 

performed by Drs. B. Ulland, R. Schueler, R. Ball, and R. Cardy.
 

The lesions of the rats and mice were reviewed by Dr. D. G.
 

Fairchild (D? and the diagnoses included in this report
 

represent his interpretations.
 

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

Mason Research Institute (3). Statistical analyses were
 

ill
 
performed by Dr. J. R. Joiner (4) and Ms. P. L. Yong (4), using
 

methods selected for the bioassay program by Dr. J. J. Gart (5).
 

The chemicals used in this bioassay were analyzed at FCRC by Dr.
 

W. Zielinsky (1). The chemical analyses were reviewed and
 

approved by Dr. W. Lijinski (1).
 

This report was prepared by Tracer Jitco (4) under the direction
 

of NCI. Those responsible for the report at Tracer Jitco were
 

Dr. C. R. Angel, Acting Director of the Bioassay Program; Dr. S.
 

S. Olin, Deputy Director for Science; Dr. J. F. Robens, toxicologist; Dr. R. L. Schueler, pathologist; Dr. G. L. Miller, Ms. L.
 

A. Waitz, Ms. M. S. King, and Mr. W. D. Reichardt, bioscience
 

writers; and Dr. E. W. Gunberg, technical editor, assisted by Ms.
 

Y. E. Presley.
 

The following scientists at NCI (2) were responsible for evaluating the bioassay experiment, interpreting the results, and
 

reporting the findings: Dr. Kenneth C. Chu, Dr. Cipriano Cueto,
 

Jr., Dr. J. Fielding Douglas, Dr. Richard A. Griesemer, Dr.
 

Thomas E. Hamm, Dr. William V. Hartwell, Dr. Morton H. Levitt,
 

Dr. Harry A. Milman, Dr. Thomas W. Orme, Dr. Sherman F. Stinson,
 

Dr. Jerrold M. Ward, and Dr. Carrie E. Whitmire.
 

(1)	 Frederick Cancer Research Center, P.O. Box B, Frederick,
 

Maryland.
 

(2)	 Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

(3)	 EG&G Mason Research Institute, 1530 East Jefferson Street,
 

Rockville, Maryland.
 

(4)	 Tracer Jitco, Inc., 1776 East Jefferson Street, Rockville,
 

Maryland.
 

(5)	 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.
 

iv
 
SUMMARY
 

A bioassay of phthalic anhydride for possible carcinogenicity was
 

conducted by administering the test chemical in feed to F344 rats
 

and B6C3F1 mice.
 

Groups of 50 rats of each sex were administered phthalic anhydride
 

at one of two doses, either 7,500 or 15,000 ppm, for 105 weeks.
 

Matched controls consisted of 20 untreated rats of each sex. All
 

surviving rats were killed at the end of the period of administration of the test chemical.
 

Groups of 50 mice of each sex were administered the test chemical
 

at one of two doses, initially either 25,000 or 50,000 ppm, for
 

32 weeks. Because of excessive depressions in the amount of body
 

weight gained in the dosed groups, the doses for the males were
 

then reduced to 12,500 and 25,000 ppm, respectively, and the
 

doses for the females were reduced to 6,250 and 12,500 ppm.
 

Administration of the test chemical at the lowered doses was
 

continued for 72 weeks. The time-weighted average doses for the
 

males were either 16,346 or 32,692 ppm, and those for the females
 

were either 12,019 or 24,038 ppm. Matched controls consisted of
 

20 untreated mice of each sex. All surviving mice were killed at
 

the end of the period of administration of the test chemical.
 

Mean body weights of the high-dose male rats and of the low- and
 

high-dose mice of each sex were lower than those of the corresponding controls; mean body weights of the low-dose male rats and
 

of both the low- and high-dose female rats were essentially
 

unaffected by administration of the test chemical. Depressions
 

in the amount of body weight gained in the male and female mice
 

were dose related throughout the bioassay. Survivals of the rats
 

and mice were not affected by administration of the test chemical.
 

No tumors occurred in the rats or mice of either sex at incidences that could be clearly related to the administration of the
 

test chemical.
 

It is concluded that under the conditions of this bioassay, 

phthalic anhydride was not carcinogenic for F344 rats or B6C3F1 

mice of either sex. 

v
 
vi

 
TABLE OF CONTENTS
 

Page
 

I. Introduction 1
 

II. Materials and Methods 5
 

A. Chemical 5
 

B. Dietary Preparation 5
 

C. Animals 6
 

D. Animal Maintenance 7
 

E. Subchronic Studies 9
 

F. Chronic Studies 12
 

G. Clinical and Pathologic Examinations 12
 

H. Data Recording and Statistical Analyses 15
 

III. Resul ts - Rats 21
 

A. Body Weights and Clinical Signs (Rats) 21
 

B. Survival (Rats) 21
 

C. Pathology (Rats) 24
 

D. Statistical Analyses of Results (Rats) 25
 

IV. Results - Mice 29
 

A. Body Weights and Clinical Signs (Mice) 29
 

B. Survival (Mice) .' 29
 

C. Pathology (Mice) 32
 

D. Statistical Analyses of Results (Mice) 33
 

V. Discussion 35
 

VI. Bibliography 37
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms in Rats
 

Administered Phthalic Anhydride
 

in the Diet 39
 

Table Al Summary of the Incidence of Neoplasms in Male
 

Rats Administered Phthalic Anhydride
 

in the Diet 41
 

vii
 
Page 

Table A2 Summary of the Incidence of Neoplasms in Female 

Rats Administered Phthalic Anhydride 

in the Diet 45 

Appendix B Summary of the Incidence of Neoplasms in Mice 

Administered Phthalic Anhydride in the Diet .. 49 

Table Bl Summary of the Incidence of Neoplasms in Male 

Mice Administered Phthalic Anhydride in the 

Diet 51 

Table B2 Summary of the Incidence of Neoplasms in Female 

Mice Administered Phthalic Anhydride in the 

Diet 54 

Appendix C Summary of the Incidence of Nonneoplastic 

Lesions in Rats Administered Phthalic Anhydride 

in the Diet 59 

Table Cl Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Administered Phthalic 

Anhydride in the Diet 61 

Table C2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Administered Phthalic 

Anhydride in the Diet 68 

Appendix D Summary of the Incidence of Nonneoplastic 

Lesions in Mice Administered Phthalic Anhydride 

in the Diet 75 

Table Dl Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Administered Phthalic 

Anhydride in the Diet 77 

Table D2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Administered Phthalic 

Anhydride in the Diet 81 

Appendix E Analyses of the Incidence of Primary Tumors 

in Rats Administered Phthalic Anhydride 

in the Diet 87 

viii
 
Page 

Table El Analyses of the Incidence of Primary Tumors 

in Male Rats Administered Phthalic Anhydride 

in the Diet 89 

Table E2 Analyses of the Incidence of Primary Tumors 

in Female Rats Administered Phthalic Anhydride 

in the Diet 94 

Appendix F Analyses of the Incidence of Primary Tumors 

in Mice Administered Phthalic Anhydride 

in the Diet 99 

Table Fl Analyses of the Incidence of Primary Tumors 

in Male Mice Administered Phthalic Anhydride 

in the Diet 101 

Table F2 Analyses of the Incidence of Primary Tumors 

in Female Mice Administered Phthalic Anhydride 

in the Diet 104 

TABLES 

Table 1 Phthalic Anhydride Subchronic Feeding Studies 

in Rats and Mice 10 

Table 2 Phthalic Anhydride Chronic Feeding Studies 

in Rats 13 

Table 3 Phthalic Anhydride Chronic Feeding Studies 

in Mice 14 

FIGURES 

Figure 1 Growth Curves for Rats Administered Phthalic 

Anhydride in the Diet 22 

Figure 2 Survival Curves for Rats Administered Phthalic 

Anhydride in the Diet 23 

Figure 3 Growth Curves for Mice Administered Phthalic 

Anhydride in the Diet 30 

IX
 
Page 

Figure 4 Survival Curves for Mice Administered Phthalic 

Anhydride in the Diet 31 

x
 
I. INTRODUCTION
 

Phthalic anhydride (CAS
 

85-44-9; NCI C03601) is an
 

important chemical intermediate
 

in the plastics industry. From
 

it are derived numerous 

Phthalic anhydride 

phthalate esters that function 

as plasticizers in synthetic
 

resins (Knuth, 1973; Noller, 1966). Phthalic anhydride itself is
 

f
 

used as a monomer for synthetic resins such as glyptal, the alkyd
 

resins, and the polyester resins (Noller, 1966). Phthalic
 

anhydride is a precursor of anthraquinone, phthalein, rhodamine,
 

phthalocyanine, fluorescein, and xanthene dyes (Towle et al.,
 

1968; Noller, 1966). Reaction of phthalic anhydride with ammonia
 

yields phthalimide, a useful reagent in the synthesis of primary
 

amines, the agricultural fungicide phaltan, and thalidomide
 

(Noller, 1966). Other reactions yield phenolphthalein, benzoic
 

acid, phthalylsulfathiazole (an intestinal antimicrobial agent),
 

and terephthalic acid (Towle et al., 1968; Noller, 1966).
 

The oral cn of phthalic anhydride for rats (strains not
 

specified) has been reported as 800-1,600 mg/kg body weight
 

(Fassett, 1964) and as 4,020 mg/kg body weight (NIOSH, 1976); the
 
LDc0 of the test chemical for white mice (route of
 

administration and strain of mouse not specified) has been
 

reported as 2,210 mg/kg body weight (Zhilova and Kasparov,
 

1968). Vapors of phthalic anhydride administered to rats over a
 

period of 12 days caused irritation of mucous membranes of the
 

nasal cavity and the bronchi (Policard et al., 1949). Persons in
 

factories manufacturing phthalic acid and phthalic anhydride can
 

develop conjunctivitis and also irritation of the skin and of
 

mucous membranes of the respiratory tract (Baader, 1955;
 

Merlevede and Elskens, 1957).
 

Phthalic anhydride was studied in the Carcinogenesis Testing
 

Program because of its high volume of production. Domestic
 

production of phthalic anhydride rose from 458 million pounds
 

annually in 1963 (Noller, 1966) to 902 million pounds in 1976
 

(United States International Trade Commission, 1977a), with
 

imports accounting for an additional 31 million pounds in the
 

latter year (United States International Trade Commission,
 

1977b). There is evidence that human exposure to phthalic
 

anhydride may occur not only in the manufacture of
 

phthalate-derived products but also in the use of plastics from
 

which phthalate plasticizers are leached, specifically certain
 

medical plastics such as blood bags, plastic syringes, and
 
plastic tubing (Guess et al., 1967). Furthermore, some phthalate
 

esters have been identified as environmental pollutants (Giam et
 

al., 1978).
 

II. MATERIALS AND METHODS
 

A. Chemical
 

Phthalic anhydride was obtained from Koppers Co. as a white, 

granular solid. The material had a melting point of 131 C 

(literature: 130.8 C). Elemental analysis showed 64.8% carbon, 

2.7% hydrogen, and 0.0% nitrogen (theoretical: 64.9%, 2.7%, and 

0.0%). Its infrared spectrum was consistent with its chemical 

structure, and identical with that of an authentic standard. The 

purity of the material was estimated by high-pressure liquid 

chromatography to be 98.8%, with one impurity. 

B. Dietary Preparation 

Test diets containing phthalic anhydride were prepared fresh every 

1 to 1-1/2 weeks in 6- to 12-kg batches at appropriate doses. A 

known weight of the chemical was first mixed with an equal weight 

of autoe laved Wayne® Sterilizable Lab Meal with 4% fat (Allied 

Mills, Inc., Chicago, 111.) using a mortar and pestle. The mixing 

was continued with second and third additions of feed, and final 

mixing was performed with the remaining quantity of feed for a 
minimum of 15 minutes in a Patterson-Kelly twin-shell blender. The 

diets were routinely stored at 5 C until used. Analyses by the 

Frederick Cancer Research Center indicated that when phthalic 

anhydride was mixed with Lab Meal at a concentration of 15,000 ppm 

and stored at room temperature for 2 weeks, the loss was 2.59% 

(372 ppm) per day. 

C. Animals 

Male and female F344 rats and B6C3F1 mice were obtained through
 

contracts of the Division of Cancer Treatment, National Cancer
 

Institute, from the NCI Frederick Cancer Research Center animal
 

farm, Frederick, Maryland as 4-week-old weanlings, all within 3
 

days of the same age. The animals were housed within the test
 

facility for 2 weeks and then were assigned four rats to a cage
 

and five mice to a cage on a weight basis for a given species and
 

sex. For use in the chronic study, the male rats were required to
 

weigh 90 to 105 g, averaging at least 100 g; the female rats, 80
 

to 95 g, averaging at least 90 g; the male mice, 18 to 22 g, averaging at least 19.5 g; and the female mice 17 to 21 g, averaging
 

at least 18.5 g. Individual animals were identified by ear punch.
 
D. Animal Maintenance
 

The animals were housed in polycarbonate cages (Lab Products,
 

Inc., Garfield, N.J.), 19 x 10-1/2 x 8 inches for the rats and
 

11-1/2 x 7-1/2 x 5 inches for the mice. The cages were suspended
 

from aluminum racks (Scientific Cages, Inc., Bryan, Tex.) and
 

were covered by nonwoven polyester-fiber 12-mil-thick filter
 

paper (Hoeltge, Inc., Cincinnati, Ohio). The bedding used was
 

Absorb-dri hardwood chips (Northeastern Products, Inc.,
 

Warrenburg, N.Y.). The feed supplied was presterilized Wayne
 

Sterilizable Lab Meal provided ad libitum in suspended stainless
 

steel hoppers and replenished at least three times per week.
 

Water, acidified to pH 2.5, was supplied ad libitum from glass
 

bottles. Sipper tubes (Lab Products, Inc.) were suspended
 

through the tops of the cages.
 

The contaminated bedding was disposed of through an enclosed
 

vacuum line that led to a holding tank from which the bedding was
 

fed periodically into an incinerator. The cages were sanitized
 

twice per week and the feed hoppers twice per month at 82 to
 

88 C in a tunnel-type cagewasher (industrial Washing Corp.,
 

Mataway, N.J.), using the detergents, Clout (Pharmacal
 

Research Laboratories, Greenwich, Conn.) or Oxford D'Chlor
 

(Oxford Chemicals, Atlanta, Ga.).
 
The glass bottles and sipper tubes were sanitized at 82 to 88 C
 

in a tunnel-type bottle washer (Consolidated Equipment Supply
 

Co., Mercersburg, Pa.) three times per week, using a Calgen
 

Commercial Division detergent (St. Louis, Mo.). The racks for
 

the cages were sanitized at or above 82 C in a rack washer
 

(Consolidated Equipment Supply Co.) once per month, using the
 

Calgen Commercial Division detergent, and the filter paper was
 

changed at the same time.
 

The air in the animal rooms was maintained at a temperature of 22
 

to 24 C and a relative humidity of 45 to 55%. Fresh air was
 

passed through a filter of 65% efficiency and a bag filter of 95%
 

efficiency at the intake and through a "Z"-type roughing filter
 

of 30% efficiency and a bag system of 90 to 95% efficiency at the
 

exhaust (American Air Filters, Louisville, Ky.; Mine Safety
 

Appliances, Pittsburgh, Pa.) and was not recirculated. The rate
 

of movement allowed 15 changes of room air per hour. The air
 

pressure was maintained negative to a clean hallway and positive
 

to a return hallway. Fluorescent lighting was provided automatically on a 12-hour-per-day cycle.
 

Both control and dosed rats were housed in the same room as rats
 

on feeding studies of the following chemicals:
 
(CAS 95-80-7) 2,4-diaminotoluene
 

(CAS 95-53-4) o-toluidine hydrochloride
 

Both control and dosed mice were housed in the same room as mice
 

on feeding studies of the following chemicals:
 

(CAS 103-33-3) azobenzene
 

(CAS 72-56-0) p,p'-ethyl-DDD
 

(CAS 20941-65-5) ethyl tellurac
 

(CAS 298-00-0) methyl parathion 

(CAS 51-03-6) piperonyl butoxide 

(CAS 88-06-2) 2,4,6-trichlorophenol 

(CAS 128-66-5) C. I. vat yellow 4 

E. Subchronic Studies
 

*
 

Subchronic feeding studies were conducted to estimate the maximum
 

tolerated doses (MTD's) of phthalic anhydride, on the basis of
 

which two concentrations (hereinafter referred to as "low" and
 

"high" doses) were selected for administration in the chronic
 

studies. Groups of five rats and five mice of each sex were
 

administered feed containing phthalic anhydride at one of several
 

doses, and groups of five control animals of each species and sex
 

were administered basal diet only. The period of administration
 

of the test chemical was 7 weeks, followed by 1 week of further
 

observation. Each animal was weighed twice per week. Table 1
 

shows the doses used and the mean body weights of dosed animals
 

at week 7 expressed as percentages of the mean weights of the
 

controls; no animals died during the Subchronic tests.
 

9
 
Table 1. Phthalic Anhydride Subchronic Feeding 

Studies in Rats and Mice 

Mean Weight at Week 7 as Percent of Control 

Dose 

(ppm) Male Female 

RATS 

6,200 90 95 

12,500 95 93 

25,000 92 91 

50,000 74 76 

MICE 

6,200 114 100 

12,500 113 99 

25 ,'000 111 101 

50,000 104 99 

10
 
At the end of the Subchronic studies, all animals were killed
 

using C0? inhalation and necropsied. The lowest dose at which
 

histopathologic findings were observed in male and female rats
 

was 25,000 ppm. At this dose, trace amounts of centrilobular
 

cytoplasmic vacuolation were seen in the livers of four males;
 

however, tissues were essentially normal in both males and
 

females at 50,000 ppm. Tissues were essentially normal also in
 

male and female mice at 50,000 ppm.
 

Ten percent depression in body weight was taken as the major
 

criterion for the estimation of MTD's. The doses required to
 

produce this response were determined by the following procedure:
 

first, least squares regressions of mean body weights versus days
 

)
 

on study were used to estimate mean body weights of each of the
 

dosed groups at day 49. Next, probits of the percent weights of
 

the dosed groups at day 49 relative to weights of corresponding
 

control groups were plotted against the logarithms of the doses,
 

and least squares regressions fitted to the data were used to
 

estimate the doses required to induce 10% depression in weight.
 

Based on these data, the low and high doses for the chronic
 

studies using male and female rats were set at 7,500 and 15,000
 

ppm. For mice, the low dose was set at 25,000 and the high dose
 

at 50,000 ppm, the maximum amount allowed for use in the
 

Carcinogenesis Testing Program.
 

11
 
F. Chronic Studies
 

The test groups, doses administered, and durations of the chronic
 

feeding studies are shown in tables 2 and 3. Because of excessive depression of the amount of body weight gained in the dosed
 

mice, doses for the low- and high-dose groups were reduced after
 

week 32 as indicated.
 

G. Clinical and Pathologic Examinations
 

All animals were checked twice daily for deaths. Observations
 

for sick, tumor-bearing, and moribund animals were recorded daily.
 

Clinical examination and palpation for masses were performed each
 

month, and the animals were weighed at least once per month.
 

Moribund animals and animals that survived to the end of the
 

bioassay were killed using C0_ and necropsied. Necropsies were
 

also performed on all animals found dead, unless precluded by
 

autolysis or severe cannibalization.
 

The pathologic evaluation consisted of gross- and microscopic
 

examination of major tissues, major organs, and all gross
 

lesions. The tissues were preserved in 10% neutral buffered
 

formalin, embedded in paraffin, sectioned, and stained with
 

12
 
Table 2. Phthalic Anhydride Chronic Feeding Studies in Rats
 

Phthalic 

Sex and Initial Anhydride Time on 

Test No. of in Diet(b) Study 

Group Animals(a) (ppm) (weeks) 

Male 

Matched-Control 20 0 105 

Low-Dose 50 7,500 105 

High -Dose 50 15,000 105 

Female
 

Matched -Control 20 0 105
 

Low-Dose 50 7,500 105
 

High-Dose 50 15,000 105
 

(a) All animals were 6 weeks of age when placed on study.
 

(b)	 Test and control diets were provided ad libitum 7 days per
 

week.
 

13
 
Table 3. Phthalic Anhydride Chronic Feeding Studies in Mice
 

Phthalic 

Sex and Initial Anhydride T ime on Time-Weighted 

Test No. of in Diet(b) Study Average Dose(c) 

Group Animals (a) (ppm) (weeks) (ppm) 

Male 

Matched-Control 20 0 104 

Low-Dose 50 25,000 32 

12,500 72 16,346 

High-Dose 50 50,000 32 

25,000 72 32,692 

Female 

Matched-Control 20 0 104 

Low-Dose 50 25,000 32 12,019 

6,250 72 

High-Dose 50 50,000 32 

12,500 72 24,038 

(a) All animals were 6 weeks of age when placed on study.
 

(b) Test and control diets were provided ad 1ibiturn 7 days per week.
 

(c) Time-weighted average dose = z(dose in ppm x no. of weeks at that dose)
 

£(no.of weeks receiving each dose)
 

14
 
hematoxylin and eosin. The following tissues were examined
 

microscopically: skin, lungs and bronchi, trachea, bone marrow
 

(femur), spleen, lymph nodes (mesenteric and submandibular),
 

thymus, heart, salivary glands (parotid, sublingual, and
 

submaxillary), liver, pancreas, esophagus, stomach (glandular and
 

nonglandular), small and large intestines, kidney, urinary
 

bladder, pituitary, adrenal, thyroid, parathyroid, pancreatic
 

islets, testis, prostate, mammary gland, uterus, ovary, brain
 

(cerebrum and cerebellum), and all tissue masses. Peripheral
 

blood smears also were made for all animals, whenever possible.
 

A few tissues from some animals were not examined, particularly
 

from those animals that may have died early, been missing, or
 

been in advanced states of cannibalization or autolysis. Thus,
 

the number of animals from which particular organs or tissues
 

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
 

15
 
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.
 

These data were analyzed using the appropriate 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 methods for testing for a dose-related trend.
 

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

16
 
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 (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 

17 
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 relationship. 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
 

18
 
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 noted,
 

in the direction of a positive dose relationship. Significant
 

departures from linearity (P less than 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 pf/p where p is the true
 

binomial probability of the incidence of a specific type of tumor
 

in a dosed group of animals and p 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
 

19
 
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
 

significant result (P less than 0.025 one-tailed test when the
 

control incidence is not zero, P less than 0.050 when the control
 

incidence is zero) has occurred. When the lower limit is less
 

than unity, but the upper limit is a 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.
 

20
 
III. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

The mean body weights of the high-dose male rats were lower than
 

those of the corresponding controls from week 13 to the end of
 

the bioassay; mean body weights of the low-dose males and both
 

the low- and high-dose females were essentially unaffected by
 

administration of the test chemical (figure 1). Arched back,
 

rough hair coat, ulceration, and corneal opacity occurred only in
 

dosed groups, but at low incidences. Wasting and tissue masses
 

were common to the dosed and control groups. Fluctuation in the
 

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

diminishes, the mean body weight may be subject to variation.
 

B. Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female rats administered phthalic anhydride
 

in the diet at the doses of this bioassay, together with those of
 

the matched controls, are shown in figure 2. The result of the
 

Tarone test for dose-related trend in mortality is not significant
 

21
 
D 9 8 

8 « B 5 8 @ i ° A 

A A A A 

A 

O 

I

o 

u! 

 300­

^ Q 

O 

m 

z 200­

e 1001 I 

D

O

A

MALE RATS 

 MATCHED CONTROL 

 LOW DOSE 

 HIGH DOSE 

40 50 60 70 

TIME ON STUDY (WEEKS) 

S3 

UI 

D 

ffl 

> 

a B o 

m B 8 8 

ilBi 

FEMALE RATS 

D MATCHED CONTROL 

O LOW DOSE 

A HIGH DOSE 

TIME ON STUDY (WEEKS) 

Figure 1. Growth Curves for Rats Administered Phthalic Anhydride in the Diet 

22 
^ 

0.90 • 

0.80 ­

_! 0.70 • 

U_ 

0

 0.50 ­

t 

5 

CO 

o 

ccp^ 

000 ­

0.90­

_l °'7° " 

< 

> 

<f> 

U. 

> 

^Jto „,.. PROBA 

3 O C J W { 3 O c 

MALE RA1 •s 

c] MATCHED CO 

CD LOW DOSiE 

^ HIGH DOSE 

fee" -••-••-••-••-..­

FEMALE RX vrs 

3 MATCH ED CO 

CD LOW DOSE 

1 HIGH DOSE 

A-"" < 3

o, 

)

Gj A-..; 

| 

45 60 75 

TIME ON STUDY (WEEKS) 

4 •••OrV"""* |^^f * 

"rW&JW 

Foi 

TIME ON STUDY (WEEKS) 

Figure 2. Survival Curves for Rats Administered Phthalic Anhydride in the Diet 

23 
in either sex. In male rats, an indicated departure from linear
 

trend (P = 0.037) is observed, due to the earlier mortality of
 

the control group when compared with that of either the high- or
 

low-dose group. The results of the Cox test applied to any two
 

of the three groups show no statistically significant difference
 

between groups of any pair.
 

In male rats, 36/50 (72%) of the high-dose group, 44/50 (88%) of
 

the low-dose group, and 14/20 (70%) of the control group lived to
 

the end of the bioassay. In females, 41/50 (82%) of the
 

high-dose group, 42/50 (84%) of the low-dose group, and 17/20
 

(85%) of the control group lived to the end of the bioassay.
 

Sufficient numbers of rats of each sex were at risk for the
 

development of late-appearing tumors.
 

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.
 

24
 
By inspection, there appeared to be no difference between the
 

dosed and control groups in frequency or distribution of
 

neoplasms, except for malignant lymphoma in the female rats. The
 

incidence of malignant lymphoma in the control females was 1/20;
 

in low-dose females, 11/50; in high-dose females, 4/50. Due to
 

the high and fluctuating incidence of this type of malignant
 

lymphoma in control F344 rats, the apparent differences in
 

incidences of the tumor in the dosed and control groups were not
 

considered to be compound related.
 

Severe chronic inflammatory, degenerative, or proliferative
 

lesions frequently seen in aged rats occurred with approximately
 

equal frequency and severity in the dosed and control groups of
 

animals.
 

Based on the histopathologic examination, there was no conclusive
 

evidence for the carcinogenicity of phthalic anhydride in F344
 

rats under the conditions of this bioassay.
 

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
 

25
 
least two animals of one group and at an incidence of at least 5%
 

in one or more than one group.
 

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

positive dose-related trend in the incidence of alveolar/
 

bronchiolar adenomas is significant (P = 0.020), but the results
 

of the Fisher exact test are not significant. The results of the
 

statistical tests on the incidences of alveolar/bronchiolar
 

carcinomas and of alveolar/bronchiolar adenomas or carcinomas are
 

not significant. In male rats, the results of the statistical
 

tests on the incidences of lung tumors are not significant.
 

A departure from linear trend (P = 0.019) is found in the
 

incidence of lymphoma in female rats, due to the relatively large
 

proportion of 11/50 (22%) in the low-dose group compared with
 

4/50 (8%) in the high-dose group and 1/20 (5%) in the control
 

group. The results of the Fisher exact test are not
 

significant. Current historical records at this laboratory
 

indicate an incidence of lymphoma in female rats of 14/285
 

(4.9%), and, although the majority of the control groups had
 

incidences of less than 5%, one control grout) was observed to
 

have an incidence as high as 4/20 (20%). Since the results of
 

the Fisher exact test were not significant and since the
 

historical data concerning lymphoma indicates the possibility of
 

26
 
an occasional high spontaneous rate of lymphoma, the evidence of
 

association of the lymphomas in the dosed group of female rats
 

with the chemical is questionable.
 

A significant dose-related trend (P = 0.037) in the negative
 

direction is observed in the incidence of pheochromocytomas of
 

the adrenal in male rats.
 

In each of the 95% confidence intervals for relative risk, shown
 

in the tables, one is included; this indicates the absence of
 

significant positive results. It should also be noted that each
 

of the intervals has an upper limit greater than one, indicating
 

the theoretical possibility of the induction of tumors by 

phthalic anhydride, which could not be detected under the 

conditions of this test. 

27
 
28

 
IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

Mean body weights of dosed male and female mice were lower than
 

those of corresponding controls throughout the bioassay, and
 

depressions in the amount of body weight gained were dose related
 

(figure 3). Tissue masses were observed at low incidences and
 

were common to the dosed and control groups. Fluctuation in the
 

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

diminishes, the mean body weight may be subject to variation.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female mice administered phthalic anhydride
 

in the diet at the doses of this bioassay, together with those of
 

the matched controls, are shown in figure 4. The result of the
 

Tarone test for dose-related trend in mortality is not
 

significant in either sex.
 

j
 

In male mice, 47/50 (94%) of the high-dose group, 37/50 (74%) of
 

29
 
O GO O O 

g 

>.

Q 

O 

m 

 30­ nB8

°

AA

HQ5A 

00

DO

 A 

o 

A 

A 

A AA . A

A AA 

5 20-i 

MALE MICE 

D MATCHED CONTROL 

O LOW DOSE 

A HIGH DOSE 

10 20 30 40 50 60 70 90 100

TIME ON STUDY (WEEKS) 

O % O 00 

A 

:oDocP AA 

o of °°aP 

< 

FEMALE MICE 

D MATCHED CONTROL 

O LOW DOSE 

A HIGH DOSE 

10 20 30 10 60 70 80 90 100 110 

TIME ON STUDY (WEEKS) 

Figure 3. Growth Curves for Mice Administered Phthalic Anhydride in the Diet 

30 

 110 
PROBABILITY OF SURVIVA

PROBABILITY OF SURVIVAL 

CD1 

*» > O D 

CO 

i m 

1 I! 

c 

i 

CA 

—** O 

> 

a. 

5" 

» •I CO 

-n 

CD 

ffin. \^r 

CD 

D 

S 
the low-dose group, and 17/20 (85%) of the control group survived
 

to the end of the bioassay. In females, 40/50 (80%) of the
 

high-dose group, 45/50 (90%) of the low-dose group, and 16/20
 

(80%) of the control group survived to the end of the bioassay.
 

Sufficient numbers of mice of each sex were at risk for the
 

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.
 

Several chronic inflammatory, degenerative, or proliferative
 

lesions frequently seen in aged laboratory mice occurred with
 

approximately equal frequency and severity in the dosed and
 

control groups of animals.
 

Based on the histopathologic examinations, the nature, incidence,
 

or severity of the lesions observed provided no clear evidence of
 

carcinogenic effect of the phthalic anhydride on B6C3F1 mice
 

under the conditions of this bioassay.
 

32
 
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 of one group and at an incidence of at least 5%
 

in one or more than one group.
 

The results of the Cochran-Armitage test for positive dose-


related trend in incidences of tumors and those of the Fisher
 

exact test comparing the incidence of tumors in the control group
 

with that in each dosed group in the positive direction are not
 

significant in either sex.
 

In male mice negative results are observed in the incidence of
 

alveolar/bronchiolar carcinomas. A significant dose-related
 

trend in the negative direction (P = 0.025) is also observed in
 

the incidence of adenomas of the thyroid in the female mice.
 

In each of the 95% confidence intervals for relative risk, shown
 

in the tables, the value of one or less than one is included;
 

this indicates the absence of significant positive results. It
 

should also be noted that each of the intervals (except for that
 

of the incidence of alveolar/bronchiolar carcinomas of the lung
 

in low-dose male mice) has an upper limit greater than one,
 

33
 
indicating the theoretical possibility of the induction of tumors
 

by phthalic anhydride, which could not be detected under the
 

conditions of this test.
 

34
 
V. DISCUSSION
 

Mean body weights of the high-dose male rats and of the low- and
 

high-dose mice of each sex were lower than those of the
 

corresponding controls; mean body weights of the low-dose male
 

rats and of both the low- and high-dose female rats were
 

essentially unaffected by administration of the test chemical.
 

Depressions in the amount of body weight gained in the male and
 

female mice were dose related throughout the bioassay. Other
 

clinical signs were common to dosed and control groups of the
 

rats and mice or occurred only at low incidences. Survivals of
 

the rats and mice were not affected by administration of the test
 

chemical. Assays of the dosed feed mixtures indicated that they
 

may have been unstable under the conditions of use.
 

In the female rats, alveolar/bronchiolar adenomas occurred at
 

incidences that were dose related in the positive direction (P =
 

0.020), but, in direct comparisons, were not significantly higher
 

in either of the dosed groups than in the control group (controls
 

0/20, low-dose 0/50, high-dose 5/50). Neither these adenomas in
 

the high-dose female rats nor any tumors in the dosed groups of
 

male rats or male or female mice can be clearly related to
 

administration of the test chemical.
 

35
 
It is concluded that under the conditions of this bioassay,
 

phthalic anhydride was not carcinogenic for F344 rats or B6C3F1
 

mice of either sex.
 

36
 
VI. BIBLIOGRAPHY
 

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

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

Baader, E. W., Erbrankungen durch Phthalsaure und ihre
 

Verbindungen. Arch. Gewerbepathol. Gewerbehyg. L3:419-453, 1955.
 

Berenblum, I., ed., Carcinogenicity Testing! A Report o£ the
 

Panel on Careinogenicity of_ the Cancer Research Commission of the
 

UICC, Vol. 2_. International Union Against Cancer, Geneva, 1969.
 

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

Soc. B 34_:187-220, 1972.
 

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

1970, pp. 48-52.
 

Fassett, D. W., Organic acids, anhydrides, lactones, acid halides
 

and amides, thioacids. In: Patty, F. A., ed., Industrial
 

Hygiene and Toxicology, Vol. II, Interscience Publishers, New
 

York, 1963T p. 1824-1825.
 

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

significance tests, confidence limits and adjustments for
 

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

Giam, C. S., Chan, H. S., Neff, G. S., and Atlas, E. L.,
 

Phthalate ester plasticizers: a new class of marine pollutant.
 

Science 199:419-421, 1978.
 

Guess, W. L., Jacob, J., and Autian, J., A study of polyvinyl
 

chloride - blood bag assemblies. Drug Intelligence
 

1^:120-121,125-127, 1967.
 

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

incomplete observations. J_. Am. Statist. Assoc. 5_3_:457-481, 1958.
 

Knuth, C. J., Plasticizers. In: Encyclopedia £f_ Chemistry,
 

Hampel, C. A. and Hawley, G. G., eds., Van Nostrand Reinhold Co.,
 

New York, 1973, pp. 863-865.
 

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

Peters, J. A., Carcinogenesis bioassay data system. Comp. and
 

Biomed. Res. 7:230-248, 1974.
 

37
 
Merlevede, E. and Elskens, J., Les intoxications dues a 1"
 

anhydride phtalique, 1' anhydride maleique et aux phtalates.
 

Arch. Bel. Med. Soc. 1M10):445-457, 1957.
 

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


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

National Institute for Occupational Safety and Health, Suspected
 

Carcinogens - A Subfile of the Registry of Toxic Effectsof
 

Chemical Substances, National Institute for Occupational Safety
 

and Health, Cincinnati, Ohio, 1976, p. 136.
 

Noller, C. R., Aromatic carboxylic acids and their derivatives.
 

In: Chemistry of Organic Compounds, W. B. Saunders Co.,
 

Philadelphia, 196T", pp. 602-605.
 

Policard, A., Gauthier, G., Hugonnier, R., and Roche, L., 

L'Intoxication par I1anhydride phtalique. Arch. Mai. Profess. 

10:1, 1949. 

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.
 

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

Biometrika 62; 679-682, 1975.
 

Towle, P. H., Baldwin, R. H., and Meyer, D. H., Phthalic acids
 

and other benzenepolycarboxylic acids. In: Kirk-Othmer
 

Encyclopedia of Chemical Technology, Vol. 15, Mark, H. P.,
 

McKetta, J. J,, Jr., Othmer, D. F., eds., John Wiley & Sons,
 

Inc., New York, 1968, pp. 444-487.
 

United States International Trade Commission, Synthetic Organic
 

Chemicals - United States Production and Sales,1976,USITC
 

Publication 833, United States International Trade Commission,
 

Washington, B.C., 1977a.
 

United States International Trade Commission, Imports of Benezoid
 

Chemicals and Products, 1976. USITC Publication 8"28,United
 

States International Trade Commission, Washington, D.C., 1977b.
 

Zhilova, N. A. and Kasparov, A. A., Phthalic anhydride and
 

N-nitrosodiphenylamine (Vulcalent A). Chem. Abstr. 71:280, 1969.
 

38
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET
 

39
 
40

 
TABLE A1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS 

ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

ANIHAIS INITIALLY IN STUDY 20 50 50
 

ANIBA1S KiCECESIEC 20 50 50
 

ANIBJLS EXAMINED HISTOPATHOIOGICALLY 20 50 50
 

IHIEGUHENIABY SYSTEM
 

*SOBCUT TISSUE (20) (50) (50)
 

EASAL-CBIL CABCINOMA 1 (2%)
 

TBICHCEEIIHELIOHA 2 (H%)
 

fIBBCSABCCMA 1 (5S) 1 (2%)
 

IIECKA 2 (H%)
 

HEHA-NGICMA 1 (2%)
 

8EUBILEHCMA, MALIGNANT 1 (2%)
 

BESFIBATCEY SYSTEM
 

#tUNG (20) (50) (50)
 

CMBCIKCMA, NOS, METASTATIC 1 (2%)
 

SCU»MOUS CELL CARCINOMA 1 (2»)
 

AlViCLAB/EfiONCHIOLAB ADiHOMA 1 (5X) 1 (8%) 1 (2%)
 

HEHATOFCIETIC SYSTEM
 

*HUITIPIE CBGANS (20) (50) (50) 

HALIGNAKT LYKPHCHA, NOS 4 (20*) 10 (20%) 12 (24%) 

BALIG.LYBEHGMA, HISTIOCYTIC TYPE 1 (2*) 

KYEICKONCCYTIC L£UKEMIA 1 (5%) 

KCNOCYTIC LEUKEMIA 1 (2%) 

*BLCCD (20) (50) (50) 

IEUKE«IA,NCS 1 (2«> 1 (2%) 

RCNCCYTIC LiiUKEHIS 1 (2X) 

*ECNE MAEEOW (20) (49) (49) 

EHAEDOBYCSAECCHA, HETASTATIC 1 (255) 

CIBCUIA1CEY SYS1I M 

IHEAR T (20) (50) (50)

 

BtiAEDOMYCSABCCH A 

* NUfBEB CF ANIMAL S fcilTH TISSUE EXAMINE D MICROSCOPICALLY 

* NUKEE B OF ANIMAL S NECEOPSIED 

41 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

MATCHED
 

CONTROL LOWD OSE HIGH DOSE
 

DIGESTIVE SYSTEM
 

*LIVEB (20) (50) (49)
 

NEOPLASTIC NODULE 1 (5%)
 

HEPATCCELLUL-AS CARCINOMA 2 (4%)
 

*DUCDENUM (20) (50) (48)
 

AEENOCAHCINCMA, 80S 1 (2%)
 

UEINAEY SYSTIE
 

NCNE
 

ENBCCFINE SYSTEM
 

tPITUITAEY (20) (49) (49)
 

CAECINCMA,NCS 2 (4%)
 

ADENCKA, NOS 5 (25%) 13 (27%) 12 (24%)
 

tADBENAL (20) (48) (49)
 

COETICAI CARCINOMA 1 (2%)
 

FHEOCHEOMCCYTOMA 6 (30%) 8 (17%) 5 (10%)
 

tTKYBOID (20) (50) (48)
 

AEENCCAECINOKA, NCS 1 (2%)
 

C-CELL AEENCM A 3 (15X) 3 3 (6%)
 

#PABATHYBOIE (17) (43) (43)
 

AEENOMA, SOS 1 (2%)
 

IPANCBEATIC ISLETS (20) (50) (49)
 

ISLET-CELL ADENOMA 2 (4%)
 

REPECEUCTIVE SYSTEM
 

*MAMMAB Y GLAND (20) (50) (50)
 

PIBBCMA 4 (8%) 1 (2%)
 

LIPOSAECOMA 1 (2%)
 

EIBECAEEKCMA 1 (2%)
 

*PEEPUTIAL GIAND (20) (50) (50)
 

CABCINCHA.NOS 1 J2%1_
 

t SOfiEEB CF ANIMALS WITH TISSUE EXAHINED HICKOSCCPICAL1-Y
 

* NDHEEE OF ANIMALS NECBCFSIiD
 

42
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

AEENCCABCINOBA, NOS 1 (2%)
 

mSTIS (20) (50) (48)
 

INTEBSTITIAI-CE1L 1UBOE 13 (65%) 40 (SOS) 35 (73X)
 

*EPIDIDYHIS 120) (50) (50)
 

1IPOHA 2 (tX)
 

NIBVCUS SYSTEM
 

IEBAIN (20) (49) (49)
 

CABCISOBA, NCS, IHYASIVE 1 (2X)
 

SPECIAL SEliSE CBGANS
 

*EYE (20) (50) (50)
 

SQOAMCUS CEIL CARCINOMA 1 (5%)
 

BUSCUICSKEIITAL SYSTEM
 

*SKUII (20) (50) (50)
 

CSTEOSAfiCOMA 1 (2X)
 

BODY CAVI1IES
 

*PEBITOKECH (20) (50) (50)
 

SAECCHA, NOS 1 (2%)
 

*1UNICA VAGINALIS (20) (50) (50)
 

KISCTHELICHA, NOS 1 (5X)
 

All C1HIE SYSTEMS
 

*BDL1IP1E CBGANS (20) (50) (50)
 

fIBECSABCCHA 1_I2X1_
 

t NDKBEB CF ANIMALS «I1H TISSUE EXAMINED BICHOSCOPICALLY
 * NDBEEE CF ANIMA1S NECEOESI2D
 

43
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

ANIMAL DISPOSITION SUMMARY
 

AKIBAIS IKITIALLY IK STUDY
 

NATUBAL DEATHS
 

MORIBUND SACEIFICE
 

SCHEDULED SACBIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACBIFICE
 

ANIMAL MISSING
 

INCLUDES AUTCLYZED ANIMALS
 

IOMOB SUMMABY
 

MATCHED
 

CONTROL
 

20
 

3
 

3
 

TCTAL ANIKAIS WI1H ESIMAfiY IUHOES* 19

TOTAL PEIMAHY TUMCBS 37

TOTAL ANIMALS HITH BENIGN TUMORS 18

TOTAL BENIGN TUflCBS


TOTAL ANIMALS WITH MALIGNANT TOMCBS


TOTAL MALIGNANT TUMOBS


TOTAL ANIMALS KITH SECONDARY TUMOBSt


TOTAL SECONDAHY TUMOBS


TOTAL ANIHALS HITH TUMOBS UNCEBTAINBENIGN OB MALIGNANT


TCTAL UNCEETAIN TUHOHS


TOTAL AKIflALS BITH TUMOBS UNCEBTAIN­ PBIMABY OB METASTATIC
 

TCTAL ONC-EfiTAIN TUflOBS
 

 28


 7


 7


 2
 

2
 

LOW DOSE HIGH DOSE 

50 50 

9 

5 

44 36 

 47

 101

 45

 77

 20

 24

 46 

 84 

 43 

 63 

 21 

 21 

3 

3 

* PBIMABY 3UMOBS: ALL TUMORS EXCEPT SECONDABY TOMOBS
 

f SECCNDABY TUMOBS: HE1ASTATIC TUMOBS OB TUHORS INVASIVE INTO AN ADJACENT OBGAN
 

44
 
T

 

TABLE A2. 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

ANIHAIS INITIALLY IN STDDY 20 50 50
 

ANIHJIS KECBCESIEE 20 50 50
 

ASIHAIS EXAMINED HISTOfATHOICGICALLY 20 50 50
 

IHTEGUBEN7ABY SYSTEM
 

*SKIN (20) (50) (50)
 

1BICHOEPITHELIOHA 1 (5X)
 

*SUECOT aiSSUE (20) (50) (50)
 

SQUAUOUS CELL CARCINOMA 1 (2%)
 

BHAEDOHYOSABCOHA 1 (2%)
 

CSTECSAECCBA 1 (5X)
 

HISFIBATCBY SYSTEH
 

*1DNG (20) (50) (50)
 

A1VECLAE/EBOKCHIOIAE ADENOMA 5 (10%)
 

A1VECLAE/BEONCHICLAE CARCINOMA 1 (5%) 3 (6%) 1 (2%)
 

COBTICJL CABCINOMA, HETASTATIC 1 (2X)
 

HIMA1CPCIE11C SYSTEM
 

*BUL1IPLE CBGANS (20) (50) (50)
 

MALIGNANT LYMPHOBA, NOS 1 (5X) 10 (20X) 4 (8%)
 

*HEBIASTINAL L.NODE (20) (50f (50)
 

MALIGNANT LYEPRORA, NOS 1 (2%)
 

CIBCUIA1CEY SYSTEM
 

NCKE
 

DIGESTIVE SYS1EM
 

fLIVEB (20) (50) (50)
 

HIPAIOCEILULAB CAECINCHA 1 (2X1
 

• NUBBEB CP ANIMALS 8ITH TISSUE EXAMINED HICBOSCOPICALLY
 * MOHEEB OF ANIMALS KECEOPSIED
 

45
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE 

(20) (50) 

1 (2%) 

(20) (50) 

1 (2%) 

11 (55%) 18 (36%) 

(20) (49) 

1 (2%) 

(20) (49) 

2 (4%) 

(20) (50) 

2 (10%) 12 (24%) 

(20) (50) 

1 (2%) 

(20) (50) 

1 (2%) 

(19) (47) 

 POLYP 1 (5%) 3 (6%) 

(20) (50) 

HIGH DOSE
 

DSINAEY SYSTEM
 

*OEINAEY ELAEDEB
 

PAPI1LCMA, NOS
 

ENECCEINI SYSTEH
 

tPHOITAEY
 

CAKCINC«A,NOS
 

AEEKCKA, NOS
 

fACBENAL
 

COBTICAL ADENOHA
 

COBTICAL CABCINOBJ
 

EHECCHBCKCCYTCHA
 

ITHYBOID
 

C-CELL ADENOMA
 

BEPECCUCTIVE SYSTEM
 

*MAHMABY GLAND
 

ADENOMA, NCS
 

CYSTADENOBA, NOS
 

FIBBCADENCMA
 

*PBEPUIIAL GLAND
 

CABCINOKA,NCS
 

*CLIIOBAL GLAND
 

ACENCHA, NCS
 

J01EBUS
 

ENDCBETEIAL STROMAL


CAECINOSABCOMA
 

8£EVCUS SYSTEM
 

tEEAIN
 

(47)
 

(49)
 

2


19


(49)
 

1


3


(50)
 

3


(50)
 

1


6


(50)
 

(50)
 

(50)
 

6


1


(50)
 

CARCINOMA, NOS, INVASIVE
 1


MIDULLCBLASTCMA 1 (2%)
 

SPECIAL SENSE CBGANS
 

NCNE
 

f NUKEEB OF JNIMALS HITH TISSUE EXAEINED HICRCSCCPICAL1Y
 * NUBBEE OF ANIMALS NECBOFSIED
 

 (4%)
 

 (39%)
 

 (2%)
 

 (6%)
 

 (6%)
 

 (2%)
 

 (12%)
 

 (12%)
 

 (2%)
 

 (2%)
 

46
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

MUSCUICSKEIEIA I SYSIE K 

NCK E 

BCE Y CAVITIE S 

NON E 

A L L CTHE K SYSTEM S 

NCN I 

ANIMA L DISPOSITIO N SUPflAE Y 

ANIKAL S INITIALL Y IK STUD Y 20 50 50 

NATUBA L D£ATH5 > 2 6 2 

MCEIEUN D SACHIFICE 1 2 7 

SCHEEULE C SACfilFICE 

ACCIDENTALL Y KILLED 

TERMINA L SACBIFICE 17 42 41 

AKIKA L KISSING 

31 INCLDDES AUTCLYZE D ANIMAL S 

# NUBBE E CF ANIMAL S WIT H TISSUE EXAMINE D HICBOSCOPICALLY 

* NUMEE B OF ANIMAI S NECEOFSIE D 

47 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

TUHOB SOBBAB Y 

TC1AL AKJHAI S HUH EEIBABY 1UHOES* 13 37 36

 

TCTAL PBIBABY TUBOtS 18 58 53

 

TOTAL ANIHAIS KITH BENIGN TDHCBS 12 27 32

 

TOTAL EESIGN TUHOJ5S 15 38 44

 

TOTAL ANIMAL S HUH MALIGNAN T TDHOES 3 16 8 

ICT6L MALIGNANT TDHOES 3 20 9

 

TOTAL ANIBA1S ilTH SECONDARY TDHCES* 1 1
 

1CTAL SECONDAHY TOHOBS 1 1
 

TOTAL AKZHALS NITH TOHOBS UNCEBTAINBEN1GN OB HALIGNANT
 

TCTAI UNCEBTAIN IUHOHS
 

TOTAL ANIHALS HUH TUHOBS UNCEBTAINP£IBAEY CB OGTASTATIC
 

TOTAL UNCERTAIN TUHORS
 

* PBIHARY 10HOBS: AIL TOHOBS EXCEPT SBCONDABY TUHORS
 

t SECCNDABY TUMORS: ME1ASTATIC TUBOBS OB TDHOBS INVASIVE INTO AN ADJACENT OBGAN
 

48
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE ADIMINISTERED PHTHALIC ANHYDRIDE IN THE DIET
 

49
 
50

 
TABLE B1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET

 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

ARIBA1S INITIALLY IN STUDY 20 50 50
 

SNIHJLS BISSMG 1
 

ANIBAIS NfeCBCESIED 20 50 49
 

ABIBilS EXAMINED HISTOPATHOLOGICAIL! 20 50 49
 

INTBGUHEN-IABY SYSTEM
 

NONE
 

HESEIBAlCBY SYSTIB
 

*LUNG (20) (50) (49)
 

HEPATOCELLULAB CABCINOHA. BEIAST 1 (5%)
 

AIVECLfiE/EBOSCHICLAE ADENOHA 2 (10S) 4 (8%) 3 (6%)
 

A1VECLAB/BEONCHIOLAB CABCINOHA 6 (30%) 2 (HI) 6 (12X)
 

HEMATCPOIETIC SYSTEM
 

*nULTIELE ORGANS (20) (50) (49)
 

BALIGNANT LYKPHOBA, MOS 3 (6X) 1 (2%)
 

«SPL£EN (19) (49) -(49 ) 

HEHANGIOM A 2 (4%) 

HEBANGIOSABCCM A 1 (5X) 1 (2%) 

iBEEENTEBIC L. NODE (20) (47) (49) 

BALIG.LYHFHOBA , HISTIOCYTIC TYPE 1 (2%) 

CIBCDIAICFY SYSTEM
 

NCNE
 

DIGESTIVE SYSTEM
 

*LIV£B (20) (50) (49)
 

HEPATOCEIIUIAB CABCINCHA 3 (15%) 12 (24%) 7 (14%)
 

# NUBBEfi CF ASIBALS VilTH TISSUE EXABINED BICROSCOPICALLY
 

* NDKEEB CF AN1HAIS NECBOPSIED
 

51
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

HEflANGICSABCCM A 2 (4%) 

OBIKAB Y SYSTE K 

*KIENE Y (20) (50) (49) 

IIPOHA 1 (2%) 

EKECCSIN I SYSTE M 

fACBENAL (19) (49) (48)
 

PEECCH80MCCYTOMA 1 (2X)
 

EiPICEUClIV£ SYSTEM
 

NONE
 

HBBVCUS SYS1IB
 

NONE
 

SPECIAL SENS2 OBGANS
 

*EYE/LACBIMAI GLAND (20) (50) (49)
 

ADENOMA, BOS 1 (2X)
 

MUSCDICSKEIEIAI SYS1EK
 

NONE
 

BODY CAVITIES
 

HCNi
 

AIL C1HEE SYS1EBS
 

*HUITIPLE OBGANS (20) (50) (49)
 

CABCINCBA, NOS^HEl ASIATIC
 

* NUBBEB CF ANIMALS HIIH TISSUE EXAMINED BICBOSCOPICALLY
 

* NOHEEB OF ABIHALS NECBOPSIED
 

52
 
TABLE 81. MALE MICE: NEOPLASMS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

SABCOHA, NO5 

ANIBA I EISECSITICK SUflHABY 

ASIBALS INITIALLY IN STUDY 20 50 50
 

NATURA L DiATHi 

HOBIBDND SACBIFICE 

SCHEDULED SACBIFICE 

ACCIEENTAILY KILLED 6
 

1EBHINAL SACRIFICE 17 37 47
 

AKIBAL MISSING 1
 

a INCLUDES ADTOLYZEC ANIHALS
 

TOHOB SCHHABi
 

TOTAL ANIHAIS HITH ERIHABY TUHOBS* 11 21 18
 

50TA1 FBIHABX TUHOBS 13 26 21
 

TOTAL ANIHAIS KITH BENIGN TUMOBS 2 6 5
 

TOTAL BBNI6N TUHOBS 2 6 6
 

TC1AL ASIBALS WITH BAIIGNANT TUHOBS 10 16 15
 

TOTAL MALIGNANT 1UMOES 11 20 15
 

TOTAL ANIHALS WITH SECONDARY TUMOBSt 1 1
 

TOTAL SfCCNDABY TUHOBS 1 1
 

TOTAL AKIBALS WITH TUMOBS UNCEETAINBENIGN OB MALIGNANT
 

TOTAL UNCEBTAIN 1UBCBS
 

TOTAL ANIHALS WITH 1UHOBS UNCEBTAIN­ PBIBABY CB HE1ASTA1IC
 

TOTAL UNCEBTAIN TUHOBS
 

* PBIHABY 1U«OBS: ALI TUHORS EXCEPT SECONDARY TUHOBS
 

f SECCNCABY 1UMOBS: HETASTATIC TUHOBS OB TOHORS INVASIVE INTO AN ADJACENT ORGAN
 

53
 
TABLE B2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET

 

ANIMAL S IMTIALLY IN £1UDY 

ANIMAL S MISSIN G 

ANIMAL S NECBCFSIEC 

ANIHAL S EXAMINE D HISTOPATHOLOGICALL Y 

INliGCHINIAfi Y SYSIEM 

*SUBCU1 TISSUE 

NEDBILEMCMA , MALIGNAN T 

BESEIFATCB Y SYSTEM 

ttiUNG 

AIVECLAB/EBCNCHIOIA B ADENOM A 

AIVECLAB/EBONCHIOLA B CAECINOM A 

CSTECSAECCMA , MEIASTATIC 

HEBATCPCIHT1 C SYS1EH 

*HUL1IPIE OEGAN S 

MALIGNAN T LYMPHCKA , NO S 

MALIG.LYMPHOMA , HISTICCYTIC TYPE 

PLASMA-CELL 1QHOE 

*BLCCD 

LEUKEMIA , NOS 

*HEflATOPOIETIC SYSTEM 

HALIGliANT LYBFHOMA , NOS 

ISPLEEN 

HEHANGIOM A 

HEMANGICSABCOM A 

IHESBNTEEIC L. NODE 

MALIGNAN T LYHPHOMA , NCS 

HSLIG.LYMPHOMA , HISTIOCYTIC 1YPE 

*LI VEE 

BALIG.LYEEHOMA , HISTIOCJTIC TYPE 

MATCHED 

CONTROL LOW DOSE 

20 50 

1 

20 49 

20 U9 

(20) (49) 

1 (5X) 

(20) (49) 

3 (6%) 

1 (5«) 3 (6X) 

(20) (49) 

1 (5%) 3 (6%) 

1 (5X) 2 (4X) 

(20) (49) 

(20) (49) 

1 (2%) 

(20) (48) 

1 (5X) 

(19) (49) 

1 (5X) 

1 (2«) 

(20) (48) 

1 12%} 

HIGH DOSE 

50 

1 

48 

48 

(48) 

(48) 

1 (2») 

1 (2%) 

1 (2% ) 

(48) 

4 (8%) 

3 (6%) 

1 (2%) 

(48) 

2 (4%) 

(48) 

(48) 

1 (2%) 

1 (295) 

(47) 

(48) 

* NUBEEB CF ANIMALS HUH TISSUE EXAHINED MICBOSCOPICALLY
 * NUMEEB OF ANIMALS NECEOPSIED
 

54
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

MATCHEP
 

CONTROL LOW DOSE HIGH DOSE
 

(20) (49) (48)
 

BALIG.LYBPHOHA, HISTIOCYTIC 1YPE 2 (4X)
 

•KIDNEY	 (19) (48) (48)
 

BALIG.LYBPHOBA, HISTIOCYIIC TYPE 1 (2X)
 

(18) (42) (37)
 

HA1IGNANT IYBPHOBA, NCS 1 (2X)
 

CIBCDIATCEY SYSTEB
 

NONE
 

DIGISlIlli SYSTEfi
 

*IIVER (20; (48) (48)
 

HiPATOCEIlULAH CAECINCBA 1 (5S) 1 (2%)
 

*CECUB (20) (49) (48)
 

LEIOHYCSAECOMA 1 (2%)
 

OBINABY SYSTIB
 

NCNE
 

ENDOCBINE SYSTEM
 

#PITUITABY (19) (46) (41)
 

ADiSCMA, NOS 1 (2%)
 

ITJiYBOID (19) (48) (46)
 

ACENOBA, NOS 2 (11X)
 

REPBCEOCTIV1 SYS3EM
 

*MABBAEY GLAND (20) (49) (48)
 

AEENCCABCINOMA, NOS 2 (4X)
 

#UTEBUS (19) (48) (46)
 

EAPIHABY CYSTAEENCCAECINOMA^NOS 1 (2*1
 

* NDCBEE CF AKIMALS KI1H 1ISSOE EXAMINED BICBOSCOPICALLY
 * NUHEEE CF ANIMAIS NECBOPSIED
 

55
 
TABLE 82. FEMALE MICE: NEOPLASMS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

ENDOMEtBIA L S1RCHAI PCIYE 1 (5X) 

• OVAB Y (18) (48) (47) 

PAPILLAR Y CYSTADEKCHA , NOS 1 (2%) 

1ERA1CMA, NOS 1 (2%) 

NtBVCU S SYS5E H 

SCN E 

SPECIAL S£^Sf C8GAN S 

*EYI/LACBIHAL GLAND (20) (US) (48) 

AEENCMA , NOS 1 (2%) 

FAPILLABY ADENOCABCINOBA 1 (2%) 

aUSCDIOSKELElAL SYSTEK 

*BCNE (20) (49) (48)
 

CSTEOSABCGHA 1 (2%)
 

BCEY CAVITIES
 

*AEBCMINAL CAVITY	 (20) (49) (48)
 

CSTECSAECCHA, ISVISIV£	 1 (2%)
 

ALL CTHBE SY2TEKS
 

*HUL1IPLE CBGANS (20) (49) (48)
 

SABCOHA, NCS 1 (5X)
 

ANIHAL DISPOSITION SUHHABY
 

ANIMALS INITIALLY IK STUDY 20 50 50
 

NATDBAL DEATH3 4 4 8
 

MCEIBUNE SACRIFICE
 

SCHEBDLED SACRIFICE
 

ACCIEENTALLY KILLED 1
 

TERMINAL SACRIFICE 16 45 40
 

AfilHAI HISSING 1 1
 

9 INCLUDES ACTOLYZEC ANIHAtS
 

* SOMBER OF ANIMALS HUH TISSOE EXAMINED MICROSCOPICALLY
 * NOHEER OF ANIMALS KECHOPSIBD
 

56
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

TOKC8 SDHHABY
 

TCTAI ANIBALS HUB PBIHAHY TUHCBS*


TOTAL PBI'HABY TOBOBS


TOTAL ANIHAIS BITH BENIGN TOHOBS


ICTAL BEKIGN TOHOBS


TOTAL ANIMALS WITH HALIGHANT TOMOBS


10TA1 HALIGBANT TOHOBS


MATCHED 

CONTROL LOW DOSE HIGH DOSE 

 10 21 17 

 11 2H 19 

3 4 3 

3 5 3 

7 16 14 

8 18 15 

TOTAL ANIHALS HUH SBCONDABY TOBOBS* 

TOTAL SECONDABX TOHOBS 

1 

2 

TOTAL AKIHJLS HITH TOHOBS ONCEBTAIH­ EENIGN OB MAIIGNANT 

TOTAL ONCEBTAIN TUHOBS 

1 

1 

1 

1 

TOT8L ANIHSLS HITH TOHOBS ONCEBTAIN^ 

PBI8ABY CB MSTASTATIC 

TOTAL UNCEBTAIN TOHOBS 

* PBIBABY TOHOBS: ALL TOHCBS EXCEPT SECONDABY TUH08S * SECONDARY TUMOBS: HE5ASTATIC TOHOBS OB TOHOBS INVASIVE INTO AH ADJACENT OBGAN
 

57
 
58

 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN RATS ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET
 

59
 
9

 0
T

 

TABLE C1. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET 

A SI«A IS IhlTIALlY IN SIUDY
 

ANIHA1S NECBOPSIEB
 

ANIBAIS EXAMINEE HISTOPATHOLOGICAL1Y
 

IKTIGUBIMTAEY SYSIZB
 

*SU5CUT TISSUE
 

INJPLAHHATICN, CBBONIC
 

KBCBCSIS, FAT
 

BESFIBATCBY SYSTEH
 

•IB ACHEA
 

INFLAHHATICN, CHEONIC
 

tXUNG
 

IDEBA, NOS
 

HEMCBBHAGE
 

ISFLAHHATICN, IN1EBSTIII AL
 

PKEUMONIA, ASPIBA7ION
 

EBCBCHCf KEOHONIA SDPPDRATIVE
 

INFLAHHA1ICS EBOLIFEBATIVB
 

FIBBCSIS
 

PEBIVASCOLITIS
 

ABTiBICSCLEBOSIS, NOS
 

tYHPHCCYtOSIS
 

HIHATCPCIE1IC SXSIEB
 

*EIOOD
 

LIUKCCYIOSIS, HOS
 

«BONE HABBOH
 

HYPEBPLASIA, HEH&IGPOIEIIC
 

•SPIEEN
 

CONGESIICH. NOS
 

MATCHED 

CONTROL 

, .,

LOM DOSE

 _ 

 HIGH DOSE 

20 

20 

20 

50

50

50

 50 

 50 

 50 

(20) (50) (50) 

1 (2X) 

1 (2X) 

(19) 

1 (5X) 

(49)

1 (2X) 

 (49) 

(20) 

1 (5X) 

16 (SOX) 

(50)

1 (2X) 

1 (2X)

1 (2X) 

1 (2X) 

2 (4X) 

39 (7SX)

 (50) 

1 (2X) 

3 (6X) 

1 (2X) 

1 (2X) 

1 (2X) 

 34 (68X) 

(20) (50) (50) 

1 (2X) 

(20) (49)

4 (8X) 

 (49) 

(20) (50)

2 (4X) 

 (49) 

« NOBBEB CF AHIHALS KITH 1ISSOE IXAHINED HICBOSCOPICALLY
 * NDBEEB OF AKIHAIS NECBOESIED
 

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

~~ '" : ~TM—=: 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

IIEBCSIS 

fIBBCSIS, FOCAL 

ISFAECT. NOS 

HEBCSICEBOSIS 

KYPCP1ASIA, NCS 

LYMEHGIE DEPLETION 

HYPEBPLASIA, B£TICULUM CELL 

HEHATOECIESIS 

GBANULCECIiSIS 

i*

10

 (20%) 

 (50%) 

1

1

12

1

1

38

1

 (2X) 

(2%) 

 (24%) 

 (2») 

 (2X) 

 (76%) 

(2%) 

1

6

1

28

 (2») 

 (12%) 

 (2%) 

 (57%) 

I1YHPH NODE 

PLASMACYTGSIS 

HYPEBPLAEIA, LYMPHOID 

(20) (50) 

1

1

 (2%) 

 (2X) 

(50) 

IHANDIEULAR I. NODE 

CYST, NOS 

CONGESTION, NOS 

EDEBA, NCS 

HEMCBEHAGE 

HiHCSIDEBOSIS 

HYPEBPLASIA, NOS 

PIASt ACYTOSIS 

(20) 

1

1

1

 (5%) 

 (5X) 

 (5X) 

(50) 

6 (12%) 

1 (2%) 

1 (2%) 

3 (6%) 

H (8%) 

(50) 

1

1

1

 (2%) 

 (2%) 

 (2%) 

SMEDIASTINAl L. NODE 

BYPEEPIASIA, LYMPHCID 

(20) (50) 

1 (2%) 

(50) 

*fliS£NlEEIC I. NODE 

CYSI, NOS 

CCNGiSlION, NOS 

HEMOSIEEECSIS 

HYPEEPIA£IA, NOS 

HYPEEPLASIA, 1YHPHOID 

(20) 

2 (10%) 

(50) 

1

1

1

1

2

 (2%) 

 (2%) 

 (2%) 

 (2%) 

 (4%) 

(50) 

1 (2%) 

IEENAL LYMPH NODE 

CONGESTION, NOS 

(20) 

1 (5X) 

(50) (50) 

•IHYHUS 

CYST, NOS 

CONGESTION, NCS 

HEBOEEHAGE 

ATBOPHY, NCS 

(13) (18) 

1

1

 (6«) 

 (6%) 

(17) 

1

1

 (6%) 

 (6%) 

CIECUIATCEY SYSTEM 

tHEABT 

JIBROSIS 

(20) 

15 (75X) 

(50) 

44 (88S) 

(50) 

43 {86jil 

* NUHBEB CF ANIHALS WITH TISSUE EXAHINED HICROSCOPICALLY * KOBEEB CP ANIHAIS BECROPSIED 

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

"' ' ~~ •^-~• "* — ^
 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

IIEBOSIS, FOCAL 1 (2X)
 

fHEAHT/ATRIUM (20) (50) (50)
 

THfiOMBCSIS, NOS 1 (2X)
 

*HYCCABDIUH (20) (50) (50)
 

FIBROSIS 1 (2X)
 

*PUIHONAEY ABTEBY (20) (50) (50)
 

HYPEBTBCEHY, NOS 1 <SX) 3 (6X) 5 (10X)
 

DIGESTIVE SYSTIfl
 

•SALIVJBY GLAND (19) (50) (50)
 

INFLAMMATION, NOS 1 (2X)
 

INFLAMMATION, CHBONIC 3 (6X) 1 (2X)
 

ILIVEB (20) (50) (49)
 

CHOLANGICFIBEOSIS 114 (70%) 42 (84X) 39 (80X)
 

NICBCSIS, NOS 1 (2X)
 

KECBCSIS, FOCAL 2 (4X)
 

BETAHCBEHCSIS FATTY 5 (25X) 10 (20X) 7 (14X)
 

LIPOIDCSIS 1 (5X) 1 (2X) 1 (2X)
 

BASOPHI1IC CYTO CHANGE 5 (25%) 20 (40X) 21 (43X)
 

FCCAI CELLULAB CHANGE 1 (2X)
 

CLEAR-CELL CHANGE 1 (2X)
 

ANGIECTASIS 1 (5%)
 

BEMATCECIISIS 1 (2X)
 

•HEPATIC CAPSULE (20) (50) (49)
 

BUPTOBE • 1 (2X)
 

JIBHCSIS, FCCAL 1 <2X)
 

ILJVEB/CENTBILOBULAB (20) (50) (49)
 

KECBOSIS, FOCAL 1 (5%)
 

*BIL£ DDCT (20) (50) (49)
 

HYPEEPLASIA, NOS 1 (5%) 3 (6X) 1 (2X)
 

#PANCBEAS (20) (50) (49)
 

CYSTIC DUCTS 1 (2X)
 

INFLAMMATION, CHECNIC 1 (5%) 1 (2%)
 

ISFLAMHATION, CHEONIC FOCAL 1 (5%)
 

FIBECSIS 4 (20X) 2 (4X) 2_(4X1_
 

# NOBBEE CF AMIMALS HITH TISSUE EXAMINED HICEOSCCPICALLY
 * NURE.EE CF AKIHALS NECBOPSIED
 

63
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED
 

CONTROL LOW DOSE
 

EEBliB-IEfilllS 1 (5X) 2 (4X)
 

ATBCPHY, NOS 4 (20X) 2 («X)
 

ISTOBACH (20) (50)
 

CIST, NOS
 

HEHATOB1, CBGANIZ5E
 

DICER, DCS 1 <5X)
 

HHfHCCYTIC IHFIAHHATOHY INFILTB
 

IIBBOSIS «» (8X)
 

IYHEHCCY1CSIS 1 (2X)
 

*GAS1EIC HUCOSA (20) (50)
 

HINEBALIZATICN
 

IGAS1RIC SUEMUCOSA (20) (50)
 

JIBBOSIS
 

•SHALL IH1ESTINE (20) (50)
 

EIVERTICOiUH 1 (2X)
 

CYST, NCS 1 (2X)
 

INFLAH0ATICN, CHflONIC FOCAL 1 (5X)
 

•COLON	 (19) (50)
 

BIBATOEIASIS
 

DfilNAEY SYSIJB 

IKIINEY 

CAST, NOS 

(20) 

15 (75X) 

(50) 

45 (90X) 

HYDECNEEHEOSIS 1 (2X) 

BEBCFBHAGE 

INFLAHHATICN, Cuov>NIC 16 (SOX) 45 (90X) 

KEEHEOEA1HY 2 (4X) 

HEHOSICfECSIS 1 (2X) 

A1BCEHY, NOS 1 (5X) 

iKIDHEY/GLODEBOLOS (20) (50) 

IJLATATICS, NCS 1 (5X) 

«KIEN£Y/IDBDIE (20) (50) 

HINEBAII2ATICN 

EILATATICN, NOS 1 (5X) 

KEPHBCSIS, NCS 

AIHCPHYiJOS
 

I NUHEEE CF ANIMALS WI1H TISSUE EXAMINED BICBOSCOPICAtLY
 

* NODEEB OF ANIMALS NECBOFSIED
 

HIGH DOSE
 

2 (4X)
 

(49)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

2 (4X)
 

1


(49)
 

1


(49)
 

1


(48)
 

(48)
 

1


(49)
 

37


1


38


1


(49)
 

(49)
 

1


1


1


 (2X)
 

 (2X)
 

 (2X)
 

 (2X)
 

 (76X)
 

 (2X)
 

 (78X)
 

 (2X)
 

 (2X)
 

 (2%)
 

 (2X1
 

64
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE 

*U. ELADDEE/BUCCS A (20) (48) 

HYPEEPLASIA , NOS 1 (2X) 

ENECCfilNE SYSTEM 

*PI3UITAEY (20) (49) 

CYST, SOS 1 (5») 5 (10X) 

IADHENA L (20) (48) 

ISFAECT, NOS 1 (2%) 

HETAHCBPHOSIS FATTY 1 (5X) 2 (US) 

HYFEBTFCEHY , FOCAL 

1ADBENAL COB1EX (20) (48) 

HYPEBTBCPHY, NOS 1 (2XJ 

HYEEBIBOEHY , FOCAL 

#ADBINAL MEDULLA (20) (48) 

HYPEKTBCEHY , NOS 

HYEEBTBOFHY, FOCAL 

HYPEBPLASIA, NOS 1 (5X) 4 (8X) 

•THYBOID (20) (50) 

HYPEHPLASIA , C-CE1L 3 (15X) 5 (10%) 

IPABATHYECI D (17) (43)

 

CYS1. NOS 

•FAKCBEA1IC ISLETS (20) (50) 

HYPEBPLASIA, NOS 2 (4X) 

HYPEBELASIA, FOCAL 

B*PECEUCTIV£ SYSTEE 

*HAMHAEY GLAND (20) (50) 

DILATATION/CUCTS 5 (25X) 12 (24X) 

*PB£PDTIAL GLAN D (20) (50) 

CYST, NOS 1 (2X) 

tPBOSTATE (20) (48)

 

CALCULUS, NCS 

i NUBBEB CF ANIMALS HUH TISSUE EXAMINE D HICBOSCOPICAILY 

* NUMEE B OF ANIMALS NECBOFSIED 

HIGH DOSE 

(48) 

(49) 

3 (6%) 

(49) 

2 (4») 

4 (8%) 

(49) 

1 (2X) 

1 (2X) 

(49) 

8 (16X) 

1 (2%) 

(48) 

10 (21X) 

(43) 

1 (2X) 

(49) 

1 (2X) 

(50) 

12 (24%) 

(50) 

(45) 

2 (4X) 

65 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

INFLAflHAlION. SUPPDBATIV£
 

ABSCESS, NOS
 

IMPtABBATICN, CHBONIC
 

IKF1ABBA1ICN, CHEONIC SUPPUBATIV
 

FIBBCSIS
 

HYEEBFLASIA, FOCAL
 

*SEHINAL VESICLE
 

INFLAMMATION, SUPPUBA1IVE
 

tTESHS
 

8EHOBBHAGE
 

IKFABCT, NOS
 

ATBCPHY, NOS
 

*JSPIDIDYMIS
 

INFLAHMATICN, CHBONIC
 

NIBVCUS SYSTEM
 

iEBAIN
 

HEMCBBHAGE
 

IKFLAMKATICN, FOCAL
 

SECBOSIS, NOS
 

SPECIAL SE1>SE CBGANS
 

NCNE
 

BUSCGICSKEIMAL £YS1EB
 

*ABECMINAL HUSCLI


HEMORRHAGE
 

EODS CAVITIES
 

*PEBI1CNEUM


HEMCBFHAGE
 

*ME£ENTEBY


PEBIAB1E BITIS
 

MATCHED 

CONTROL LOW DOSE HIGH OOSE 

2 (IX) 1 (2X) 

1 (2X) 

1 (5X) 

1 (5X) 

1 (2X) 

1 (5%) 

(20) (50) (50) 

1 (5X) 

(20) (50) (48) 

1 (2X) 

1 (2X) 

3 (6X) 2 (IX) 

(20) (50) (50) 

1 (2X) 

(20) (49) (49) 

2 (*X) 

1 (2X) 

2 (4X) 

 (20) (50) (50) 

1 (2%) 

 (20) (50) (50) 

1 (2X) 

 (20) (50) (50) 

t NUKBEB OF ASIBALS HI-IH TISSUE EXAMINED BICBOSCOPICALLY
 * HUHEEB CF ANIMALS NECBCPSIED
 

66
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

NECROSIS , FA1 1 (2%) 

ALL C1HER SYSILHS 

ADIPOSE TISSUE 

LYHPHOCXTIC INFLAHHATOBY INFILTB 

SPECIAL MORPHOLOGY SUKBARY 

NON E 

* NUKBEB 01 ANIMALS KITK TISSUE EXAEINtD BICEOSCCPICALLY * NtKEEK CF ANIMALS NECROES1ED 

67
 
TABLE C2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

ANIMALS INITIALLY IN STUDY

ANIMALS KECECPSIEE

ANIMALS EXAMINED HISICEATHCLOGICAILY

 20

 20

 20

 50

 50

 50

 50 

 SO 

 50 

INTIGliMINlARY SYS1EM 

*SKIS

OLCEE, NOS

INFLAMMATION, ACUTE

ATBCPHY, NCS

 (20) (50)

 1 (2%) 

1 (2%) 

1 (2%) 

 (50) 

*SUECDT TISSUE

THROMBOSIS, NOS

 (20) (50)

 1 (2%) 

 (50) 

BESPIEATCBY SYSTEM 

*LUNG

INFLAMMATION, INTEBSTITIAL

INFLAMMATION, GRiNULOKATOUS

ABTEBICSCLEBCSIS, NOS

IYMPHCCYTOSIS

 (20)

 2 (10X)

 1 (5%)

 17 (85S)

 (50)

 6 (12%)

 3 (6%)

 1U (88%)

 (50) 

5 (10%) 

1 (2%) 

1 (2%) 

 42 (84%) 

HIMATCPCIITIC SYSTEM 

*ELCCD 

IEUKCCYTCSIS, NOS 

(20) (50) (50) 

1 (2%) 

IBONE HARSOB 

BYPEfiPIASIA, NOS 

HYPESFLAEIA, HEflATOPOIETIC 

(19) (U8) 

3 (6«) 

(H6) 

1 (2%) 

•SPLEEN 

CCNGESTICS, NOS 

SCLEBCSIS 

FIBECSIS, FOCAL 

INFAECT, NOS 

HFJCSIEBJCSIS 

(20) 

1 (5X) 

(50) 

1 (2*) 

1 (2*) 

(50) 

2 (4%) 

32_J64«i 

t NOBEfB CF ANIMALS KITH TISSUE EXAMINED HICBOSCOPICALLY 

* HDMEEB OF ANIHALS NECBOPSIED 

68
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

BYPEBPLASIA, HlflitOPOIETIC
 

HYPEBPLiSIA, BETICULUfl CELL
 

HEH1TOPOIESIS
 

*LYHPH NODE
 

CONGESTION, NOS
 

PLASMACYIOSIS
 

HYPERPLASIA, LYHEHOID
 

tHJNDIBULAH L. NOD£
 

CIST, NOS
 

HYP2BPLASIA, NOS
 

BYPERPLASIA, BI1ICULUH CELL
 

HYPEBPLASIA, LYflFHOID
 

tflEDIASTINAL L. NODE
 

CONGESTION, NOS
 

HYPEBPLASIA, RETICUIUB CELL
 

JHESENTERIC L. NODE
 

CYST, NOS
 

CONGESTION, NOS
 

ISJIA8HATION, GBANOLOB ATOUS
 

HYPEBPLASIA, RETXCULUH CELL
 

•IHYHOS
 

ATROPHY, NOS
 

CIRCDLATCBY SYS1IB
 

tHIABl
 

FIBEOSIS
 

IYBEHCCY1CSIS
 

IHEART/A1BIOH
 

TBECMBCSIS. NOS
 

*BYOCABEIUM
 

INFLAMMATION, NOS
 

*AB1ERY
 

INFLAKMA1ICN, NOS
 

*PULHONABY AB1EBY
 

HYPEBIROEHY. NOS
 

MATCHED
 

CONTROL
 

16 (SOX)
 

(20)
 

(20)
 

1 (5X)
 

(20)
 

(20)
 

1 (5X)
 

1 (5X)
 

1 (5X)
 

(9)
 

(20)
 

13 (65X)
 

(20)


1 (5X)
 

(20)
 

(20)
 

(20)
 

1 (5XJ
 

LOW DOSE
 

1 <2X)
 

39 (78X)
 

(50)
 

1 (2X)
 

1 (2%)
 

(50)
 

1 (2%)
 

1 (2X)
 

(50)
 

3 (6X)
 

1 (2X)
 

(50)
 

1 (2X)
 

(29)
 

(50).
 34	 (68X)
 

1 (2X)
 

(50)
 

(50)
 

(50)
 

1 (2X)
 

(50)
 

HIGH DOSE
 

1 (2X)
 

45 (90 X)
 

(50)
 

2 (*X)
 

(50)
 

1 (2X)
 

1 (2X)
 

(50)
 

(50)
 

(20)
 

1 (5X)
 

(50)
 

40 (80X)
 

(50)
 

(50)
 

1 (2X)
 

(50)
 

(50)
 

* NUBBEB CF ANIBALS WI1H TISSUE EXAMINED BICBOSCOPICALLY
 

* NUHEEB OF ANIMALS NtCBOESIED
 

69
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

*H£SENT£BIC ABTEHX
 

PE8IABTEBI1IS
 

DIGES1IV! SYSTEE
 

tSAUVAB Y GLSN D 

INFLAMMATION, NOS
 

EIEBOSIS
 

OFLAHHATIGN, SUPPORATIVE
 

SESC£SS, NOS
 

GEANULCMA, NOS
 

MBBCSIS, FOCAL
 

CHOLANGICFIBBGSIS
 

CIBBHOSIS, NOS
 

KECECSIS, NOS
 

SECBCSIS, FOCAL
 

KETAHCBEHQSIS FAT1Y
 

BASOPHILIC CYTO CHANG£
 

FCCAL CEILULAB CHANGE
 

MEGALCCY1CSIS
 

IEUKCCY10SIS, NEU1EOPHILIC
 

fLIVEfi/CENTBILOEUIAE
 

SECHCSIS, NOS
 

iLI-VEE/PEBIPOBTAL
 

IIPCIEOSIS
 

tPANCHEAS
 

FIBBOSIS
 

JIBBCSIS, FCCAL
 

EEBIABTEBI1IS
 

KECBCSIS, FAT
 

ATBCEHY, NOS
 

A1BCFHY, FCCAL
 

IS10MACH
 

OLCEE, NCS
 

INFLABHAIICN, SUPFOBATIVE
 

IHFLABHATJON, CHBONIC
 

FIBBCSIS _______
 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

(20) (50) (50) 

1 (2%) 

(19) (49) (50) 

1 (2%) 

1 (2X) 

(20) (50) (50) 

1 (2X) 

1 <2X) 

1 (2%) 1 (2X) 

3 (6») 

16 (80S) 28 (56%) 38 (765?) 

1 (5X) 

1 (2*) 1 (2X) 

3 (6%) 1 (2%) 

1

17

 (5X) 

 (85*) 

3 (658) 

43 (86S) 

4 (8%) 

39 (76%) 

1 (2%) 1 (2X) 

1 (5%) 

1 <2X) 

(20) (50) (50) 

1 (2%) 

(20) (50) (50) 

1 (2%) 

(19) (49) (49) 

1 (5%) 1 (2%) 3 (6X) 

1 (2*) 

1 (2X) 

1 (2%) 

1 (5*) 2 (4*) 

1 (2X) 

(20) (50) (50) 

1 (2%) 

1 (2%) 

1 (2X) 

t NUflBEB CF ANIMALS SI1H TISSUE EXAMINED MICROSCOPICALLY
 * NUBEEH CF ANIMALS RECBOESIED
 

70
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

HYPEBPLASIA, EEIIHEIIAL 1 (2X)
 

LYHEHCCYTGSIS 2 (4X)
 

ILABGE INtESlINE (20) (49) («9)
 

NEMATODIASIS 1 (2X)
 

ICOION (20) (49) (49)
 

KJHATODIA5IS 1 (2X) 1 <2X)
 

UBIKABX SYSTEB
 

IKIENEY (20) (50) (50)
 

CAST, NOS 10 (SOX) 19 (38X) 16 (32X)
 

HYDBCNEPbBOSIS 1 (2X)
 

CONGESTION, NOS 1 (2X)
 

INFLAMMATION, INTEBSTIIIAL 1 <5X)
 

INFLAMMATION, CHEONIC 14 (70X) 42 (84%) 40 (SOX)
 

SC1EBCSIS 1 (2X)
 

HEHOSIEEBOSIS 1 (5X) 1 (2X) 1 (2X)
 

tKIDNEY/EELVIS (20) (50) (50)
 

CALCULUS, NOS 1 (2%)
 

tfOBINARY BLADDER (20) (50) (47)
 

HEMOBBHAGE 1 (2X)
 

HYPEBPLASIA, AEENCMATOUS 1 (2%)
 

•
 iNECCBINE SYS1EK
 

tPiaOIIABY (20) (50) (49)
 

CYST, NOS 5 (25X) 22 (44X) 12 (24X)
 

CCNGES'IICN, NCS 2 <4X)
 

HEMCEBHAGIC CYST 2 (10X) 2 (4%) 1 (2X)
 

INFLAMWA1ICN, OSSIFYING 1 (2%)
 

AKGIECIASIS 1 (2»)
 

4ADBENAL (20) (49) (49)
 

CONGESTION, NOS 1 (2%)
 

HEMCBBHAGIC CYS7 1 (5») 1 (2*)
 

KETAHCBPHCSIS PATTY 3 (15%) 2 (4X) 1 (2X)
 

PIGMENTATION, NOS 1 (2X)
 

flYPEBTBCEHY, NOS 2 (10S)
 

#ACBENAL COBTEX (20) (49) (49)
 

BETABCBPHOSIS FATTY 1 (2X1
 

* NOEBEB CF ANIMALS WI1H TISSUE EXAMINED MICROSCOPICALLY
 

* NUHEEB CF ANIMALS NECROPSIED
 

71
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

______— ———— —— _- — - ___— —— —— —	 --• -^--L

-L - ——• •"" '— .
 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

HYPEBTBOPHY, DOS 2 (4%) 2 (4«)
 

HYPEBTBOEHY, FOCAL 1 (5X) 1 (2X)
 

HYPESPLASIA, NOS 1 (2S)
 

•ADRENAL	 MEDULLA (20) (49) (49)
 

HYPEBPLASIA, NOS 1 (2X)
 

*THYBOID (20) (09) (50)
 

UITIMOEBANCHIAI CYSI 1 (2X)
 

fOLLICULAB CYST, NOS 1 (2K)
 

INFLAMMATION, CHBONIC FOCAL 1 (2%)
 

HYPEEPLASIA, C-CELL 1 (5«) 9 (18%) 6 (12X)
 

IPANCBEA7IC ISLETS (19) <"9) (49)
 

HYPEBPLASIA, NOS 1 (2»)
 

BEPECIUCTIVI SYSTEM
 

*MAMHABY GLAND (20) (50) (50)
 

EILATATICN/DUCTS 13 (658) 33 (66X) 24 (48S)
 

GALACTCCELE 1 (5X) 4 (8%) 1 (2*)
 

INFLAM8ATICN, GBANULOH ATOOS 1 (2%)
 

FIBSOSIS 1 (2»)
 

HYPEBPLASIA, NOS 1 (2%)
 

HYPEBPLASIA, FOCAL 1 (5X)
 

HYPEBPLASIA, CYSTIC 1 (2%)
 

tUlEBUS (19) (47) (50)
 

HAHABTCHA 1 (2X)
 

CILATA1ICS, NCS 1 (2X)
 

KECBCSIS, NOS 1 (5%)
 

•UTEBOS/ENDOMETBIOB	 (19) (47) (50)
 

DILATATION, NCS 1 (2X) 1 (2X)
 

CIS1, NOS 1 (2X)
 

HYPEBPLASIA, EPITHELIAL 1 (2X)
 

IENDOHETBIAL GLAND (19) (47) (50)
 

DILATATION, NOS 3 (16X)
 

tOVAEY (19) (47) (50)
 

CYST, NOS 1 (5X) 3 (6X) 1 (2X)
 

INFLAMMATION, CHBONIC 1 (5X)
 

HYPCPIASIA, NCS 1 (2X)
 

t NUHBEE CF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 * NUflEEB OF ANIHALS NECBOPSIED
 

72
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

ATBCPHY, NOS 1 (2%) 

NERVCU S SYSIEfl 

iLJlEBAL V£N1BIC1E (20) (50) (50) 

DILATATION, NOS 1 (5X) 

*EBAIN (20) (50) (50)
 

HINEBAIIZATICN 1 (2%)
 

II1ATATICN, NOS 1 (2X)
 

H1TDBCCEEHAI.OS, NCS 1 (2X) 1 (2%)
 

SPECIA1 S£KSE OBGANS
 

*EYE (20) (50) (50)
 

FIBBCSIS 1 (2X)
 

*EYE/CCENEA (20) (50) (50)
 

BUETUfiB 1 (2S)
 

HDSCOIOEKHElAi SYS1ER
 

*SKULL (20) (50) (50)
 

EXOSIOSIS 1 (2%)
 

BCBV CAVI3IIS
 

*PiBICAP,riDH (20) (50) (50)
 

INFLAOMATION, FIERINOUS 1 (2%)
 

All CTKEfi SYSTEMS
 

NONE
 

SEECIAI KCBEHCLCGY SUBBABY
 

KC LESICN BEPCEIEE . 1
 

* NUHBEE OF AKIMALS WITH TISSUE EXAMINED HJCBOSCOPICAILY
 * NUMEEB CF ASIHALS NECBOPSIED
 

73
 
74

 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN MICE ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET
 

75
 
76

 
TABLE 01.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET 

MATCHED 

CONTROL 

—————

LOW OOSE 

——­

HIGH DOSE 

ANieAIS IKI-UAILY IN STUDY

ANIMALS KISSING 

ANIBALS KECECPSIED

ASTHALS EXAMINED HI STCPATHOLOGICA1LY

 20 

 20 

 20 

50 

50 

50 

50 

1 

49 

49 

ISTEGOHENIAKY SYSTEM
 

*SKO (20) (50) (49)
 

AIKOPHY, NOS 1 (2«)
 

*SDECU1 TISSUE (20) (50) (49)
 

AESCESS. NOS 1 (2«)
 

BESEIBATCBY SYSTEH
 

• TBKHEA (20) (» 8) (45)
 

HEBOBBHAGE 1 (2%)
 

• LONG (20) (50) (49)
 

EEBOBEHAGE 2 (4S) 1 (2*)
 

EBONCHOENEUMONIA, NOS 1 (5X)
 

IYBEHCCY1IC INFLABBATOaY INFILTB 1 (2%)
 

INFIAHHATICN, INTEESTITIAL 1 (5%)
 

mFLAKHAHON PBOLIFEB JlIVjB 1 (2X)
 

ALVEOIAB MACKCEHAGES 1 (2X)
 

LYHFHCCY10SIS 6 (30%) 19 (38%) 30 (61%)
 

HIBATCPCII1IC SYS1EK
 

*ELCCD (20) (50) (49)
 

lEUKOCYlCSiS, NOS 1 (2X)
 

tBONE MAE ROM (20) (49) (49)
 

BYPEBPLASIA, HEHATOPOIETIC 1 12%)
 

ISPLEEN (19) (49) (49)
 

CONGESTION, NOS 1 (2%)
 

HJPEBPLASIA^ LYHEHOID 2 (4%)
 

• NUBBEB CF AMIBALS HIIH TISSUE JEXAHINED BICBOSCOPICAI1Y
 

* NUBEEB OF ARIBALS NECBOPSIED
 

77
 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

tSPIENIC FOLLICLES (19) (49) (49)
 

JHROEHY. NCS 1 (2X)
 

ILYHEH NODE (20) (47) (49)
 

HYPEBPLASIA, NOS 1 12%)
 

1HAND1EOLAB L. NODE (20) (47) (49)
 

CYSI, NCS 3 (15X)
 

HEMC8EHAGE 1 (5*)
 

ttBONCHIAL LYMPH NODE (20) (47) (49)
 

INFLAMMATION, CHECNIC 1 (5%)
 

JMESEKTEFIC L. NODE (20) (47) (49)
 

IYMPHANGIECTASIS 1 (5%)
 

1HRCHECSIS, NOS 1 (2S)
 

CCNGESTICN, NCS 3 (15X) 7 (15X) 7 (14*)
 

HYEEEPLASIA, NOS 1 <5X)
 

HYPEBPIASIA, HETICULUM CELL 4 (9%) 5 (10%)
 

HYPEEPLASIA, LYMPHOID 1 (2-X)
 

S1HYMUS (12) (38) (47)
 

CYST, NCS 1 (8%)
 

CIECOIA1CEY SYSTEM
 

IHEABT (20) (49) (49)
 

FIBEOSIS 1 (5%)
 

FIBHCSIS, FOCAL 1 (2%)
 

EEK.IAB1EBIIIS 1 (2%)
 

#HEAHT/ATEIUfl (20) (49) (49)
 

THBOKEOSIS, NCS 1 (2X)
 

IMYCCAEDIUM (20) (49) (49)
 

INFLAMMATION, CHRONIC FCCAL 1 (5%)
 

#HE£ATIC SINUSOID (20) (50) (49)
 

IEOKCCYTCSIS, NOS 1 (2*)
 

DJGISIIVE SYS1EK
 

ILIVEB (20) (50) (49)
 

FIBECSIS. FOCAL 1 (2%)
 

t NUBBEE CF ANIMALS VillH 1ISSUE EXAMINED MICEOSCOPICALLY
 

* NUMEE6 CF ANIMAIS NECBOPSIED
 

78
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED
 

CONTROL LOW DOSE
 

NECECSIS, FOCAL 2 (10%) 2 (4%)
 

ISF4BCT, NC5 1 (2%)
 

KETAMCBEHCSIS FA1IY 5 (25%) 3 (6%)
 

LIPOIDCSI3 6 (12%)
 

REGALCCYTOSIS 1 (22)
 

HYPEBPLASIA, NOLULAB 1 15%)
 

ANGIEC1ASIS i (5%) 1 (2*)
 

tEIIE DUCT (20) (50)
 

INFIAKHATICN, CHBCNIC 1 (5%) 7 (14%)
 

*PANCB£AS (20) (49)
 

FIBFCSIS
 

*S1CHACH (19) (48)
 

INFIAHBATICN, NOS 1 (2%)
 

INFLAMMATION, FOCAL 3 (6»)
 

•SHALL INTESTINE (20) (47)
 

ULCEE, NOS
 

GBANUIAIICN, TISSUE
 

iCOICN (20) (48)
 

EOLYF
 

URINABY SYS1IP 

4KJEKEY 

CAS1, SOS 

IHFLAWKATICN, CHBONIC 

AHYLOICCSIS 

IYMPHCCYTCSIS 

(20) 

1

9

 (5%) 

 (45%) 

(50) 

22

1

15

 (44%) 

 (2%) 

 (30%) 

ENECCBIKE SYSTEM
 

IADBENAL (19) (49)
 

AMYLOIDCSIS 1 (2%)
 

HYPEBTBCEHY, FOCAI 1 (2%)
 

*ADEENAL COETEX (19) (49)
 

AlRCPbY, NOS 23 (4VS)
 

HYPEBTBCEHY, NCS
 

HYfEBTBOEHY, FOCAL .
 

t NUKBEE CF AKIHALS KI1H 1ISSUE EXAMINED HICBOSCOPICALLY
 

* NUHEEB CF ANIMAIS NECBOPSIED
 

HIGH DOSE
 

1


3


(49)
 

17


(47)
 

1


(49)
 

(49)
 

1


1


(49)
 

1


(49)
 

J


37


(48)
 

(48)
 

40


1


1


 (2%)
 

 (6%)
 

 (35%)
 

 (2%)
 

 (2%)
 

 (2%)
 

 (2%)
 

 (6%)
 

 (76%)
 

 (83%)
 

 (2%)
 

 (2%)
 

79
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

BEPBCDUC1IVE SYSTfM
 

*EPIEIDYMIS


INFLAMMATION, CHRONIC


NEBVCUS SYSTEM
 

*EBAIN


CONGESTION, NOS


HEHOBBHAGE


IBBAIN/THALAMUS


MINERALIZATION


SPECIAL SISSI OBGANE
 

NONE
 

HOSC1JIOSKEIE1AL SXS1EH
 

NONE
 

BCEK CAVI1IIS
 

*HESIN3EBX


WINEBALIZATION


JECBCSIS. FAT


ALL C1HIE SYSTEMS
 

*HUI1IP1E OBGANS


LYBEhCCYTOSIS


SPECIAI KCEEBCICGY SUKPABY
 

NC LESION BEPOBTED


ANIBAL KISSING/NC NECBOPSY


MATCHED 

CONTROL LOW DOSE HIGH DOSE 

 (20)	 (50) (49)
 

1 (5%)
 

 (19) (50)	 (49)
 

1 (2%)
 

1 (5%) 1 (2%)
 

 (19)	 (50) (49)
 

 18 (36X) 23 (4755)
 

 (20)	 (50) (49)
 

1 (5%)
 

2 (10%)
 

 (20)	 (50) (49)
 

2 (<*%)
 

2 2
 

1
 

*	 NUREEB CF ANIMALS WI1H TISSUE EXAMINED MICBOSCOPICALLY
 

*	 NDHEEB CF ANIMALS NECBOFSIED
 

80
 
TABLE D2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

ANIHA1S INITIALLY IN STUDY 20 50 50 

ANIMAL S MISSING 1 1 

ANIHAIS NECBOPSIED 20 49 48 

AHIBAIS EXAMINE D HISTOPATHOIOGICALLY 20 49 48 

INTEGDBENTAEY SYS3IM
 

NONB
 

BESPI8ATCRY SYSTEM
 

4IONG (20) (49) (48)
 

INFLAMMATION, INTEESTITIAL 2 (4%)
 

LYHPHCCYTOSIS 2 (10%) 32 (65%) 34 (71%)
 

HEBAtCPCIETIC SYS1EB
 

*EIOCD (20) (49) (48)
 

IEOKCCYTCSIS, NEUTEOPHILIC 1 (5%) 1 (2%)
 

tSPIEEN (20) (48) (48)
 

HXPEBPLASIA, IYMPHOID 1 (2%)
 

HEBATOECIESIS 1 (5%)
 

iLYHPH NODE (19) (49) (47)
 

KYPEEPLASIA, NOS 1 (5%)
 

HXPEBPLASIA, IYHPHOID 1 (2%)
 

IflANDIEOLAR L. NODE (19) (49) (47)
 

HSPEBPLASIA, NOS 1 (5%)
 

HYPEBPLASIA. LYHPHCID 1 (2X)
 

IBESENIEEIC L. NODE (19) (47)
 

LYHPHANGIECTASIS 3 (6%)
 

CCNGESTICK, NOS 2 (11%) 1 (2%) 3 (6%)
 

EDEBA, NOS 1 (2%)
 

HEMOERHAGE 1 (2%)
 

ISF1ABBATICN, CHBONIC 1 J2S1
 

t HOBBES OF ANIHALS WITH TISSUE EXABINED MICROSCOPICALLY
 

* NOMBEB CF ANIMA1S NECEOPSIED
 

81
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

HYPEEELASIA, NOS 1 (2X)
 

HYPEBP1ASIA, BITICULUfl CELL 9 (18X)
 

HYEEKPLASIA, LYMEHOID 1 (2X)
 

CIBCCIATCEY SY3ZEB
 

1HEAHT (19) (.8, (47)
 

BINEBALIZATICN 1 (5X)
 

EIGJSIIVI SYSIEK
 

•LIVES	 (20) (48) (48)
 

ISMPHCCYTIC INFLABKATCBY INFILT5 1 (2X)
 

I.ECBCSIS, NOS 1 (2X)
 

NECBOSIS, FCCAL 1 (5X)
 

tETAKCEEHCSIS FATTY 1 (2X)
 

LEUKCCYTC3IS, NOS 1 (2X)
 

GBANULCECIESIS 1 (2X)
 

tLIVEE/CiNTBILOEULAR (20) (48) (48)
 

IIPOIECSI5 1 (2S)
 

JtEItE EUCT (20) (48)
 

EILATATION. NCS 1 (5X) 1 (2X)
 

INFLABKAIICN. CEEONIC 10 (SOX) 30 (63X) 36 (75X)
 

*PANCBEAS (18) (49) (47)
 

INFLAMMATION, NOS 1 (2X)
 

INFLABBATICN, CHBCNIC 1 (2*)
 

IIEECSIS 1 (2*) 1 (2S)
 

FIBECSIS, FCCAL 1 (2X)
 

AlBCtHY, NCS 1 (2X)
 

A1BCEHY, FCCAL 1 (2X)
 

*S1CHACH (20) (49) (48)
 

INFIAHMATICN, ACUTE FCCAL 1 (5X)
 

*DUCDENUM (20) (49)
 

ECLYE 1 (2%)
 

UBIKJBY SYS1EK
 

*KIENEY	 (19) (48) (48) 

IYBPJCCYIIC_IMIAKKATCBY_INFILTB LJ2SI. 

* NUBEEB CF ANIMALS kllH 1ISSUE EXAMINED HICBOSCOPICALLY
 

* NUKEEE CF ANIMALS NZCEOESIED
 

82
 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

INFLAMMATION , CKECNIC
 

LYMPHCCY1CSIS
 

HYFEEFLASIA, LYMFHCID
 

*KICNEY/TUEULE
 

NECECSIS, NCS
 

#UBINAEY ELADDER
 

ECEMA, NCS
 

*0. ELAELiR/SUbHUCCSA
 

FIBECSIS
 

EKrCCBIKH SYSTE M
 

lADRtNAL
 

CYS1, NOS
 

f!ETAMCREHCSIS FAT1Y
 

IADEENAL COETEX
 

ATRCPHY, NCS
 

HYEEFTBCEHY, FOCAI
 

tADEENAL MEDULLA
 

HYPEETECEHY, FCCA1
 

#TKYfiOID
 

FIEECSIS, FCCAL
 

A1BCEHY, NOS
 

fiEEEClUCTIVE SYS'IEH
 

IU1EEUS
 

DILATATION, NOS
 

BE£KA, KCS
 

EYCHtTEA
 

fU-IEBUS/iNDCMiJTFIOM
 

EILATATICN, NCS
 

IKFLAKMATICN, NOS
 

INFLAMMATION, CHi-CNIC
 

HYPEFFLASIA, FAPILLAEY
 

HYPEF.FLA£IAt CYSTIC
 

* NUKBE E CF AMMAL S hiaH TISSUE

* NUMEE K CF AKIKA1 S KECEOFSIE D 

MATCHED
 

CONTROL LOW DOSE
 

1 (5%) 9 (19%)
 

22 (46%)
 

(IS) (48)
 

1 (2%)
 

(18)	 (47)
 

(18)	 (47)
 

1 (6%)
 

(18)	 (46)
 

1 (2%)
 

1 (2%)
 

(18)	 (46)
 

16 (89%) 37 (80%)
 

(18)	 (46)
 

(19)	 (48)
 

(19)	 (48)
 

2	 (11X)
 

1 (2X)
 

(19) (48)
 

5	 (26%) 29 (60%)
 

1 (2%)
 

1 (2«)
 

 EXAHINIC M.ICEOSCCPICALLY 

HIGH DOSE
 

26 (54%)
 

1 (2%)
 

(48)
 

(47)
 

1 (2%)
 

(47)
 

(46)
 

(48)
 

42	 (88%)
 

1 (2%)
 

(48)
 

1 (2%)
 

(46)
 

1 (2X)
 

1 (2%)
 

(46)
 

1 (2%)
 

(46)
 

20 (43%)
 

1 (2%)
 

1 (2%)
 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

»OVAEY


CIST, NOS


HBHCBBHAGIC CYST


ATRCEHY, NOS


NSBVCOS SYS1BB
 

IBBAIN


HEHOKBHAGE


KICBCSIS, NOS


IBBfllN/IHALAMUS


HISEKALIZATI08


SPICIAI S£tlS£ CEGANS
 

NONE
 

MDSCUIOSKELEliL SYS1IK
 

*BC»E


HEALED FHACTUBE


CSTECPOBCSIS


*FEHOB


CSTEOPCBCSIS


ECEY CAVJTIBS
 

*PEEI1CNEOM


KECFCSIS, FAT


*ME££NTEEY


KECBOSIS, FAT


ALL CfHEE SYS1EHS
 

NONE
 

I NUHEEB CF ANIMAIS HI1H TISSUE


* NDKEEB CF ANIMALS NECECESIED
 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

 (18) (48) (47) 

2 (11%) 26 (54%) 7 (15%) 

1 (6%) 1 (2%) 1 (2%) 

1 (2%) 

 (20) (49) (48) 

1 (2%) 

1 (2«) 

 (20) (49) (48) 

8 (40%) 20 (41%) 26 (54%) 

 (20) (49) (48) 

1 (2%) 

4 (8%) 

 (20) (49) (48) 

1 (5%) 

 (20) (49) (48) 

1 (2%) 

 (20) (49) (48) 

2 (4X) 

 EXAMINED MICBOSCOPICAILY 

84
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

SPECIAL KCEFKCLCGY SUKHAEY 

ANIMAL HISSIKG/NC NECfiOESY

NECECESY EEBF/NO HISTC PEBFCBMEB

AUTC/NECEOESY/HIS10 P.EHF

AUTC1YSIS/NO NECKCFSY

 1 

1 

2 

1 

I NUSEEE OF ANIKAIS WITH TISSUE IXABINED MICEOSCOPICALLY 

* NCBEEE OF ANIKA1S NECEOfSIED 

85
 
86

 
APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

IN RATS ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET
 

87
 
88

 
00 

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

Administered Phthalic Anhydride in the Diet (a)
 

Matched Low
 

Topography: Morphology Control Dose
 

Lung: Alveolar /Bronchiolar
 

Adenoma (b) 1/20 (5) 4/50 (8)
 

P Values (c,d) N.S. N.S.
 

Relative Risk (f) 1.600
 

Lower Limit 0.175
 

Upper Limit 77.169
 

Weeks to First Observed Tumor 105 105
 

Hematopoietic System:
 

Leukemias (b) 1/20 (5) 1/50 (2)
 

P Values (c,d) N.S. N.S.
 

Relative Risk (f) 0.400
 

Lower Limit 0.005
 

Upper Limit 30.802
 

Weeks to First Observed Tumor 83 105
 

High
 

Dose
 

1/50 (2)
 

N.S.
 

0.400
 

0.005
 

30.802
 

105
 

3/50 (6)
 

N.S.
 

1.200
 

0.106
 

61.724
 

79
 
(continued) 

Topography;

Table El.

 Morphology

 Analyses of the Incidence of Primary Tumors in Male Rats 

Administered Phthalic Anhydride in the Diet (a) 

Matched 

 Control 

Low 

Dose 

High 

Dose 

Hematopoietic System: 

Lymphomas (b) 4/20 (20) 11/50 (22) 12/50 (24) 

P Values (c,d) N.S. N.S. N.S. 

Relative Risk (f) 

Lower Limit 

Upper Limit 

1.100 

0.384 

4.321 

1.200 

0.429 

4.650 

Weeks to First Observed Tumor 102 94 96 

vo 

O 

Hematopoietic System: 

Lymphomas or Leukemias

P Values (c,d)

 (b) 5/20 (25) 

 N.S. 

12/50

N.S. 

 (24) 15/50

N.S. 

 (30) 

Relative Risk (f) 

Lower Limit 

Upper Limit 

0.960 

0.376 

3.124 

1.200 

0.497 

3.770 

Weeks to First Observed Tumor 83 94 79 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered Phthalic Anhydride in the Diet (a)
 

(continued) 

Topography: Morphology 

Matched

Control

 Low 

 Dose 

High 

Dose 

Pituitary: Adenoma, NOS (b) 5/20 (25) 13/49 (27) 12/49 (24) 

P Values (c,d) N.S. N.S. N.S. 

Relative Risk (f) 

Lower Limit 

Upper Limit 

1.061 

0.425 

3.404 

0.980 

0.384 

3.184 

Weeks to First Observed Tumor 83 96 84 

Adrenal: Pheochromocy toma (b) 6/20 (30) 8/48 (17) 5/49 (10)
 

P Values (c,d) P = 0.037 (N) N.S. N.S.
 

Relative Risk (f) 0.556 0.340
 

Lower Limit 0.202 0.096
 

Upper Limit 1.734 1.205
 

Weeks to First Observed Tumor 89 94 96
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered Phthalic Anhydride in the Diet (a)
 

(continued)
 

Tomography: Morphology


Thyroid: C-cell Adenoma (b)


P Values (c,d)


Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Mammary Gland: Fibroma (b)


P Values (c,d)


Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched Low High 

 Control Dose Dose 

 3/20 (15) 3/50 (6) 3/48 (6) 

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

0.400 0.417 

0.060 0.062 

2.802 2.915 

 105 105 105 

 0/20 (0) 4/50 (8) 1/50 (2) 

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

Infinite Infinite 

0.386 0.022 

Infinite Infinite 

105 105 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats 

Administered Phthalic Anhydride in the Diet (a) 

(continued) 

Matched Low High 

Topography: Morphology Control Dose Dose 

Testis: Interstitial-cell Tumor (b) 13/20 (65) 40/50 (80) 35/48 (73) 

P Values (c,d) N.S. N.S. N.S. 

Relative Risk (f) 1.231 1.122 

Lower Limit 0.882 0.792 

Upper Limit 1.864 1.766 

Weeks to First Observed Tumor 74 90 84 

(a) Dosed groups received 7,500 or 15,000 ppm.
 

>£> (b) Number of tumor-bearing animals/number of animals examined at site (percent).
 

(c) Beneath the incidence of tumors in the control group is the probability level for the Cochran-


Armitage test when P is less than 0.05, otherwise, not significant (N.S.) is indicated. Beneath
 

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

for the comparison of that dosed group with the matched-control group when P is less than 0.05;
 

otherwise not significant (N.S.) is indicated.
 

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

(e) The probability level for departure from linear trend is given when P is less than 0.05 for any
 

comparison.
 

(f) The 95% confidence interval of the relative risk between each dosed group and the control group.
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Phthalic Anhydride in the Diet (a)
 

Topography: Morphology


Lung: Alveolar/Bronchiolar
 

Adenoma (b)


P Values (c,d)


Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Lung: Alveolar/Bronchiolar
 

Carcinoma (b)


P Values (c,d)


Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched
 

 Control
 

 0/20 (0)
 

P = 0.020
 

—
 

 1/20 (5)
 

 N.S.
 

105
 

Low
 

Dose
 

0/50 (0)
 

.
 

—
 

—
 

—
 

—
 

3/50 (6)
 

N.S.
 

1.200
 

0.106
 

61.724
 

101
 

High
 

Dose
 

5/50 (10)
 

N.S.
 

Infinite
 

0.525
 

Infinite
 

105
 

1/50 (2)
 

N.S.
 

0.400
 

0.005
 

30.802
 

102
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Phthalic Anhydride in the Diet (a)
 

(continued)
 

Matched Low High
 

Topography; Morphology Control Dose Dose
 

Lung: Alveolar /Bronchiolar Carcinoma
 

or Adenoma (b) 1/20 (5) 3/50 (6) 6/50 (12)
 

P Values (c,d) N.S. N.S. N.S.
 

Relative Risk (f) 1.200 2.400
 

Lower Limit 0.106 0.175
 

Upper Limit 61.724 108.021
 

Weeks to First Observed Tumor 105 101 102
 

Hematopoietic System:
 

vo Lymphomas (b) 1/20 (5) 11/50 (22) 4/50 (8)
 

in
 

P Values (c,d) N.S. N.S. N.S.
 

Departure from Linear Trend (e) P=0.019
 

Relative Risk (f) 4.400 1.600
 

Lower Limit 0.722 0.175
 

Upper Limit 184.752 77.169
 

Weeks to First Observed Tumor 101 105 102
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Phthalic Anhydride in the Diet (a)
 

(continued)
 

Topography ; Morphology
 

Pituitary: Adenoma, NOS (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Pheochromocytoma (b)
 

VO P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched Low High 

Control Dose Dose 

11/20 (55) 18/50 (36) 19/49 (39) 

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

0.655 0.705 

0.385 0.419 

1.280 1.363 

81 101 16 

0/20 (0) 0/49 (0) 3/49 (6) 

N.S. — N.S. 

Infinite 

— 0.255 

Infinite 

94 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Phthalic Anhydride in the Diet (a)
 

(continued)
 

Topography: Morphology


Thyroid: C-cell Adenoma (b)


P Values (c,d)


Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Fibroadenoma (b)


\o
 

P Values (c,d)


Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched
 

 Control
 

 0/20 (0)
 

 N.S.
 

 2/20 (10)
 

 N.S.
 

105
 

Low
 

Dose
 

2/49 (4)
 

N.S.
 

Infinite
 

0.125
 

Infinite
 

105
 

12/50 (24)
 

N.S.
 

2.400
 

0.614
 

20.902
 

101
 

High
 

Dose
 

3/50 (6)
 

N.S.
 

Infinite
 

0.250
 

Infinite
 

94
 

6/50 (12)
 

N.S.
 

1.200
 

0.243
 

11.574
 

74
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Phthalic Anhydride in the Diet (a)
 

(continued)
 

Uterus: Endometrial
 

Strotnal Polyp (b) 1/19 (5) 3/47 (6) 6/50 (12)
 

P Values (c,d) N.S. N.S. N.S.
 

Relative Risk (f) 1.213 2.280
 

Lower Limit 0.107 0.311
 

Upper Limit 62.303 102.629
 

Weeks to First Observed Tumor 105 105 105
 

(a) Dosed groups received 7,500 or 15,000 ppm.
 

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

oo
 

(c) Beneath the incidence of tumors in the control group is the probability level for the Cochran-


Armitage test when P is less than 0.05, otherwise, not significant (N.S.) is indicated. Beneath
 

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

for the comparison of that dosed group with the matched-control group when P is less than 0.05;
 

otherwise not significant (N.S.) is indicated.
 

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

(e) The probability level for departure from linear trend is given when P is less than 0.05 for any
 

comparison.
 

(f) The 95% confidence interval of the relative risk between each dosed group and the control group.
 
APPENDIX F
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

IN MICE ADMINISTERED PHTHALIC ANHYDRIDE IN THE DIET
 

99
 
100

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

Administered Phthalic Anhydride in the Diet (a)
 

Topography: Morphology

Matched 

 Control 

Low 

Dose 

High 

Dose 

Lung: Alveolar/Bronchiolar 

Carcinoma (b) 6/20 (30) 2/50 (4) 6/49 (12) 

P Values (c,d) N.S. P = 0.005 (N) N.S. 

Departure from Linear Trend (e) P = 0.007 

Relative Risk (f) 

Lower Limit 

Upper Limit 

0.133 

0.015 

0.681 

0.408 

0.129 

1.372 

Weeks to First Observed Tumor 100 97 104 

Lung: Alveolar/Bronchiolar Carcinoma or
 

Adenoma (b) 7/20 (35) 6/50 (12) 9/49 (18)
 

P Values (c,d) N.S. P = 0.032 (N) N.S.
 

Relative Risk (f) 0.343 0.525
 

Lower Limit 0.114 0.211
 

Upper Limi t 1.061 1.464
 

Weeks to First Observed Tumor 89 97 104
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered Phthalic Anhydride in the Diet (a)
 

(continued)
 

Matched Low
 

Topography: Morphology Control Dose
 

Hematopoietic System:
 

Lymph omas (b) 0/20 (0) 3/50 (6)
 

P Values (c,d) N.S. N.S.
 

Relative Risk (f) Infinite
 

Lower Limit 0.250
 

Upper Limit Infinite
 

Weeks to First Observed Tumor — 93
 

Liver: Hepatocellular
 

o Carcinoma (b) 3/20 (15) 12/50 (24)
 NJ
 

P Values (c,d) N.S. N.S.
 

Relative Risk (f) 1.600
 

Lower Limit 0.503
 

Upper Limit 8.185
 

Weeks to First Observed Tumor 104 101
 

High
 

Dose
 

2/49 (4)
 

N.S.
 

Infinite
 

0.125
 

Infinite
 

99
 

7/49 (14)
 

N.S.
 

0.952
 

0.250
 

5.317
 

104
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered Phthalic Anhydride in the Diet (a)
 

(continued)
 

(a) Dosed groups received time-weighted average doses of 16,346 or 32,692 ppm.
 

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

(c) Beneath the incidence of tumors in the control group is the probability level for the Cochran-


Armitage test when P is less than 0.05, otherwise, not significant (N.S.) is indicated. Beneath
 

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

for the comparison of that dosed group with the matched-control group when P is less than 0.05;
 

otherwise not significant (N.S.) is indicated.
 

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

(e) The probability level for departure from linear trend is given when P is less than 0.05 for any
 

comparison.
 

(f) The 95% confidence interval of the relative risk between each dosed group and the control group.
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered Phthalic Anhydride in the Diet (a)
 

Topography: Morphology


Lung: Alveolar/Bronchiolar
 

Carcinoma (b)


P Values (c,d)


Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Lung: Alveolar/Bronchiolar Carcinoma or
 

Adenoma (b)


P Values (c,d)


Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched Low 

 Control Dose 

 1/20 (5) 3/49 (6) 

 N.S. N.S. 

1.224 

0.108 

62.958 

 99 44 

 1/20 (5) 6/49 (12) 

 N.S. N.S. 

2.449 

0.332 

110.166 

99 44 

High
 

Dose
 

1/48 (2)
 

N.S.
 

0.417
 

0.006
 

32.058
 

104
 

2/48 (4)
 

N.S.
 

0.833
 

0.047
 

48.155
 

104
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered Phthalic Anhydride in the Diet (a)
 

(continued)
 

Matched Low High
 

Topography: Morphology Control Dose Dose
 

Hematopoietic System:
 

Lymphomas (b) 3/20 (5) 11/49 (22) 8/48 (17)
 

P Values (c,d) N.S. N.S. N.S.
 

Relative Risk (f) 1.497 1.111
 

Lower Limit 0.460 0.308
 

Upper Limit 7.741 6.043
 

Weeks to First Observed Tumor 104 90 71
 

Hematopoietic System:
 

Lymphomas or Leukemias (b) 3/20 (15) 11/49 (22) 9/48 (19)
 o
 

Ul
 

P Values (c,d) N.S. N.S. N.S.
 

Relative Risk (f) 1.497 1.250
 

Lower Limit 0.460 0.361
 

Upper Limit 7.741 6.662
 

Weeks to First Observed Tumor 104 90 71
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered Phthalic Anhydride in the Diet (a)
 

(continued)
 

Matched Low High
 

Topography : Morphology Control Dose Dose
 

Thyroid: Adenoma, NOS (b) 2/19 (11) 0/48 (0) 0/46 (0)
 

P Values (c,d) P = 0.025 (N) N.S. N.S.
 

Departure from Linear Trend (e) P = 0.043
 

Relative Risk (f) 0.000 0.000
 

Lower Limit 0.000 0.000
 

Upper Limit 1.329 1.386
 

Weeks to First Observed Tumor 104
 — —
 

(a) Dosed groups received time-weighted average doses of 12,019 or 24,038 ppm.
 

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

(c) Beneath the incidence of tumors in the control group is the probability level for the Cochran-


Armitage test when P is less than 0.05, otherwise, not significant (N.S.) is indicated. Beneath
 

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

for the comparison of that dosed group with the matched-control group when P is less than 0.05;
 

otherwise not significant (N.S.) is indicated.
 

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

(e) The probability level for departure from linear trend is given when P is less than 0.05 for any
 

comparison.
 

(f) The 95% confidence interval of the relative risk between each dosed group and the control group.
 
Review of the Bioassay of Phthalic Anhydride* for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

December 13, 1978
 

The Clearinghouse on Environmental Carcinogens was established
 

in May, 1976, in compliance with DREW 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 on the Institute's bioassay program to identify and evaluate
 

chemical 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, and State health officials.
 

Members have been selected on the basis of their experience in carcinogenesis or related fields and, collectively, provide expertise in
 

chemistry, biochemistry, biostatistics, toxicology, pathology, and
 

epidemiology. Representatives of various Governmental agencies participate as ad hoc members. The Data Evaluation/Risk Assessment Subgroup
 

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 in this context that the below critique is given on the bioassay of Phthalic Anhydride.
 

The reviewer for the report on the bioassay of Phthalic Anhydride
 

agreed with the conclusion that the compound was not carcinogenic under
 

the conditions of test. After a brief description of the experimental
 

design, he said that both the dose levels tested and the animal survival
 

were adequate. There was no objection to the reviewer's motion that
 

the report on the bioassay of Phthalic Anhydride be accepted as written.
 

Clearinghouse Members Present:
 

Arnold L. Brown (Chairman), University of Wisconsin Medical School
 

Joseph Highland, Environmental Defense Fund
 

William Lijinsky, Frederick Cancer Research Center
 

Henry Pitot, University of Wisconsin Medical Center
 

Verne A. Ray, Pfizer Medical Research Laboratory
 

Verald K. Rowe, Dow Chemical USA
 

Michael Shimkin, University of California at San Diego
 

Louise Strong, University of Texas Health Sciences Center
 

Kenneth Wilcox, Michigan State Health Department
 

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.
 

107
 

JJ.U.S. GOVERNMEN T PRINTIN G OFFICE : 1979-281-217/301 2
 

DHEW Publication No. (NIH) 79-1715