## NTP TECHNICAL REPORT

ON THE



# TOXICOLOGY AND CARCINOGENESIS

STUDIES OF 1- BROMOPROPANE

(CAS NO. 106

- 94 -5)



# IN F344/N RATS AND B6C3F1 MICE

(INHALATION STUDIES)

NATIONAL TOXICOLOGY PROGRAM

P.O. Box 12233

Research triangle Park, NC 27709

August 201 1

NTP TR 564

NIH Publication No. 11

- 5906



## National Institutes of Health

Public Health Service

U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES

## FOREWORD

The National Toxicology Program (NTP) is an interagency program within the Public Health Service (PHS) of the

Department of Health and Human Services (HHS) and is headquartered at the National Institute of Environmental

Health Sciences of the National Institutes of Health (NIEHS/NIH). Three agencies contribute resources to the

program: NIEHS/NIH, the National Institute for Occu pational Safety and Health of the Centers for Disease Control

and Prevention (NIOSH/CDC), and the National Center for Toxicological Research of the Food and Drug

Administration (NCTR/FDA). Established in 1978, the NTP is charged with coordinating toxicological testing

activities, strengthening the science base in toxicology, developing and validating improved testing methods, and

providing information about potentially toxic substances to health regulatory and research agencies, scientific and

medical comm unities, and the public.

The Technical Report series began in 1976 with carcinogenesis studies conducted by the National Cancer Institute.

In 1981, this bioassay program was transferred to the NTP. The studies described in the Technical Report series ar e

designed and conducted to characterize and evaluate the toxicologic potential, including carcinogenic activity, of

selected substances in laboratory animals (usually two species, rats and mice). Substances selected for NTP toxicity

and carcinogenicity s tudies are chosen primarily on the basis of human exposure, level of production, and chemical

structure. The interpretive conclusions presented in NTP Technical Reports are based only on the results of these

NTP studies. Extrapolation of these results to other species, including characterization of hazards and risks to

humans, requires analyses beyond the intent of these reports. Selection per se is not an indicator of a substance’s

carcinogenic potential.

The NTP conducts its studies in compliance with its laboratory health and safety guidelines and FDA Good

Laboratory Practice Regulations and must meet or exceed all applicable federal, state, and local health and safety

regulations. Animal care and use are in accordance with the Public Health Service Policy on Humane Care and Use

of Animals. Studies are subjected to retrospective quality assurance audits before being presented for public review.

NTP Technical Reports are indexed in the NIH/NLM PubMed database and are available free of charge

electron ically on the NTP website ( http://ntp.niehs.nih.gov ) or in hardcopy upon request from the NTP Central Data

Management group at cdm@niehs.nih.gov or (919) 541

- 3419.

## NTP TECHNICAL REPORT

ON THE



# TOXICOLOGY AND CARCINOGENESIS

STUDIES OF 1- BROMOPROPANE

(CAS NO. 106

- 94 -5)



# IN F344/N RATS AND B6C3F1 MICE

(INHALATION STUDIES)

NATIONAL TOXICOLOGY PROGRAM

P.O. Box 12233

Research triangle Park, NC 27709

August 2011

NTP TR 564

NIH Publication No. 11

- 5906



## National Institutes of Health

Public Health Service

U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES

### 2



# CONTRIBUTORS

National Toxicology Program

Evaluated and interpreted results and reported findings



### D.L. Morgan, Ph.D.

, Study Scientist



### M.F. Cesta, D.V.M.

, Study Pathologist



### J.B. Bishop, Ph.D.

J.R. Bucher, Ph.D.

R.S. Chhabra, Ph.D.

P.M. Foster, Ph.D.

D. Germolec, Ph.D.

R.A. Herbert, D.V.M., Ph.D.

M.J. Hooth, Ph.D.

A.P. King

- Herbert, D.V.M.

G.E. Kissling, Ph.D.

D.E. Malarkey, D.V.M., Ph.D.

J.H. Roycroft, Ph.D.

C.S. Smith, Ph.D.

G.S. Travlos, D.V.M.

S. Waidyanatha, Ph.D.

N.J. Walker, Ph.D.

K.L. Witt, M.S.



## Battelle Toxicology Northwest

Conducted studies and eval uated path o logy findings



### J.A. Dill

, Ph.D.

, Principal Investigator



### S.L. Grumbein, D.V.M., Ph.D.

S.J. Harbo, D.V.M.

B.K. Hayden

R.A. Renne, D.V.M.



## Experimental Pathology Laboratories, Inc.

Provided pathology review



### M.H. Hamlin, II, D.V.M., Principal Investigator



### E. T. Adams

, D.V.M., Ph.D.

R.A. Miller, D.V.M., Ph.D.

J.C. Peckham, D.V.M., M.S., Ph.D.

C.C. Shackelford, D.V.M., M.S., Ph.D. 

## TherImmune Research Corporation

Provided SMVCE analysis



### G.W. Wolfe, Ph.D., Principal Investigator



### Y. Wang, M.S.



## Dynamac Corporation

Prepared quality assessment audits



### S. Brecher, Ph.D., Principal Investigator



### S. Iyer, B.S.

V.S. Tharakan, D.V.M.



## NTP Pathology Working Group

Evaluated slides and contributed to pathology report on

2- year rats ( December 11 , 200 7)



### G.D. Hill, D.V.M., Ph.D.

, Coordinator

ILS, Inc.



### E. T

. Adams , D.V.M., Ph.D.

Experimental Pathology Laboratories, Inc.



### M.F. Cesta

, D.V.M

.

National Toxicology Program



### S.A. Elmore

, D.V.M., M.S.

National Toxicology Program



### B. Hamilton

, D.V.M., Ph.D.

GlaxoSmithKline



### R.A. Herbert

, D.V.M., Ph.D.

National Toxicology Program



### R.R. Maronpot, D.V.M

.

Consultant



### R. A. Miller, D.V.M., Ph.D.

Experimental Pathology Laboratories, Inc.



### A. Nyska, D.V.M

.

National Toxicology Program



### B. P. Singh, B.V.Sc., M.S.

National Toxicology Program

### 1- Bromopropane , NTP TR 564 3



## NTP Pathology Working Group

Evaluated slides and contributed to pathology report on

2- year mice ( February 5 , 200 8)



### K.R. Hobbie, D.V.M.

, Coordinator

ILS, Inc.



### M.F. Cesta

, D.V.M.

National Toxicology Program



### S.A. Elmore, D.V.M., M.S.

National Toxicology Program



### S.L. Grumbein

, D.V.M., Ph.D.

Battelle Toxicology Northwest



### B. Hamilton, D.V.M., Ph.D.

GlaxoSmithKline



### D.E. Malarkey, D.V.M., Ph.D.

National Toxicology Program



### R.R. Maronpot, D.V.M.

Consultant



### R.A. Miller, D.V.M., Ph.D.

Experimental Pathology Laboratories, Inc.



### J.C. Peckham, D.V.M., M.S., Ph.D.

Experimental Pathology Laboratories, Inc.



### C.C. Shackelford, D.V.M., M.S., Ph.D.

Experimental Pathology Laboratories, Inc. 

## SRA International , Inc.

Provided statistical analyses



### P.W. Crockett, Ph.D., Principal Investigator



### L.J. Betz, M.S.

K.P. McGowan, M.B.A.



## Biotechnical Services, Inc.

Prepared Technical Report



### S.R. Gunnels, M.A., Principal Investigator



### L.M. Harper, B.S.

P.C. Rathman, B.S.E.

D.C. Serbus, Ph.D.

### 4



# C ONTENTS

ABSTRACT

................................................................................................ ................................................................ . 7

EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY

........................................ 13

TECHNICAL REPORTS REVIEW SUBCOMMITTEE

..................................................................................... 14

SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMITTEE COMMENTS

................................ 15

INTRODUCTION

................................................................................................ ..................................................... 17

MATERIALS AND METHODS

.............................................................................................................................. 25

RESULTS

................................................................................................ ................................................................... 37

DISCUSSION AND CONCLUSIONS

..................................................................................................................... 73

REFERENCES

................................................................................................ .......................................................... 79

A PPENDIX 

### A Summary of Lesions in Male Rats in the 2

- Year I nhalation Study

of 1- Bromopropane

....................................................................................................................... 85

A PPENDIX 

### B Summary of Lesions in Female Rats in the 2

- Year Inhalation Study

of 1- Bromopropane

.................................................................................................................... 103

A PPENDIX 

### C Summary of Lesions in Male Mice in the 2

- Year Inhalation Study

of 1- Bromopropane

.................................................................................................................... 119

A PPENDIX 

### D Summary of Lesions in Fem ale Mice in the 2

- Year Inhalation Study

of 1- Bromopropane

.................................................................................................................... 133

A PPENDIX 

### E Genetic Toxicology

...................................................................................................................... 149

A PPENDIX 

### F Clinical Pathology Results

.......................................................................................................... 155

A PPENDIX 

### G Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios

................................................... 163

A PPENDIX 

### H Reproductive Tissue Evaluations and Estrous Cycle Characterization

................................ . 169

### 5

A PPENDIX 

### I Chemical Characterization

and Generation of Chamber Concentrations

........................................................................... 173

A PPENDIX 

### J Ingredients, Nutrient Composition, and Contaminant Levels

in NTP

- 2000 Rat and Mouse Ration

.......................................................................................... 183

A PPENDIX 

### K Sentinel Animal Program

........................................................................................................... 187

### 6 1- Bromopropane, NTP TR 564

SUMMARY

Background

1- Bromopropane is used for degreasing and as a solvent for adhesive resins. We studied 1

- bromopropane to

determine if it caused cancer in rats or mice.

Methods

We exposed groups of 50 male and female rats and mice to air containing 1

- bromopropane six hours per day, five

days a week for two years. Rats were exposed to concentrations of 125, 250, or 500 parts per million (ppm) of

1- bromopropane in air, and mice were exposed to concentrations of 62.5, 125, or 250 ppm. Similar groups of

50 animals were exposed to clean air in the same inhalation chambers as the treated control groups six hours per

day. Tissues from more than 40 sites were examined for every animal.

Results

A few rare tumors of large intestine were seen in exposed male and female rats, and a variety of skin tumors were

seen in exposed male rats and to a lesser extent in exposed female rats. Male rats also had slightly increased

incidences of malignant mesotheliomas and of pancreatic islet adenoma and carcinoma. Female mice had markedly

increased incidences of adenomas and carcinomas of the lung. Male and female rats and mice exposed to

1- brom opropane had hyperplasia of the nose. Some exposed male and female rats also had inflammation of the

larynx, and most exposed male and female mice had regeneration of bronchioles of the lung.

Conclusions

We conclude that 1

- bromopropane caused cancer of the large intestine in male and female rats and of the lung in

female mice. 1

- Bromopropane caused tumors of the skin in male rats and possibly in female rats also. Malignant

mesotheliomas and pancreatic islet adenomas and carcinomas in male rats also were possibly associated with

exposure to 1

- bromopropane. Rats and mice of both sexes exposed to 1

- bromopropane had hyperplasia and

inflammation of the nose and other effects in the upper respiratory system (inflammation of the larynx in rats,

regeneration o f lung tissue in mice).

### 7



# ABST RACT

1- BROMOPROPANE

CAS No. 106

- 94 -5

Chemical Formula: C 3 

### H 7 

### Br Molecular Weight: 12 2. 99

Synonyms : 1- BP; p ropyl bromide; n- BP; N- propyl bromide



### In the early to mid 1990s, 1

- bromopropane was used

primarily as an intermediate in the production of pesti

cides, quaternary ammonium compounds, flavors and

fragrances, pharmaceuticals, and other chemicals in

well

- controlled, closed processes. In the mid to late

1990s, it was introduced as a less toxic replacement for

methylene chloride in emissive applications such as

vapor and immersion degreasing operations and critical

cleaning of electronics and metals. 1

- Bromopropane

was also introduced as a nonflammable, nontoxic, fast

drying, and inexpensive solvent for adhesive resins, and

has been marketed as a replacement for ozone depleting

refrigerants. 1- Bromopropane was nominated for study

by the O ccupation al S af ety and H ealth A dministration

based on the potential for widespread occupational and

environmental exposure and a lack of toxicity and car

cinogenicity data. Male and female F344/N rats and

B6C3F1 mice were exposed to 1- bromopropane (99%

or greater pure) by inhalation for 2 weeks , 3 months

, or

2 years. Genetic toxicology studies were conducted in Salmonella typhimurium and Escherichia coli and

mouse peripheral blood.



# 2-W EEK 

# S TUDY IN 

# R ATS

Groups of five male and five female rats were exposed

to 1

- bromopropane vapor at concentrations of 0, 125,

250, 500, 1,000, or 2,000 ppm, 6 hours plus T 90 

### (12 minutes) per day, 5 days per week for 16 days. A ll

rats survived to the end of the study except one 500 ppm

male . Mean body weights of 2,000 ppm rats were sig

nificantly less than those of the chamber controls. The

absolute kidney weight of 1,000 ppm male s, relative

kidney weights of all exposed groups of males , and

absolute and relative kidney weights of all exposed

groups of females were significantly in creased. The

absolute and relative liver weights of 1,000 ppm males,

relative liver weights of 500 and 2,000 ppm males, and

absolute and relative liver weights of 500 ppm or

greater females were significantly increased

. Nasal

### 8 1- Bromopropane , NTP TR 564

l esions included s uppurativ e inflammation in males

exposed to 500 ppm or greater, respiratory epithelial

necrosis in 1,000 and 2,000 ppm males, and respiratory

epithelial regeneration in 1,000 and 2,000 ppm females.



# 2-W EEK 

# S TUDY IN 

# M ICE

Groups of five male and five female mice were exposed

to 1

- bromopropane vapor at concentrations of 0, 125,

250, 500, 1,000, or 2,000 ppm, 6 hours plus T 90 

### (12 minutes) per day, 5 days per week for 17 days. All

2,000 ppm males

, two 2,000 ppm females, four

500 ppm males, one 1,000 ppm male, and one

1,0 00 ppm female died early. The m ean body weight

gain of 1,000 ppm males was significantly less than that

of the chamber controls. Abnormal breathing, lethargy,

and eye discharge were observed primarily during

week 1 in groups exposed to 500 ppm or greater . Liver

weights of 1,000 ppm males and of females exposed to

500 ppm or greater were significantly increased

. Kid

ney weights of 1,000 and 2,000 ppm females were sig

nificantly increased. Microscopic lesions related to

1- bromopropane exposure occurred i n the lung, liver,

and nose of male s and females and were primarily seen

in mice exposed to 500 ppm or greater

.



# 3-M ONTH 

# S TUDY IN 

# R ATS

Groups of 10 male and 10 female rats were exposed to

1- bromopropane vapor at concentrations of 0, 62.5, 125,

250, 500, or 1,000 ppm, 6 hours plus T 90 

### (10 minutes)

per day, 5 days per week for 14 weeks. Additional

clinical pathology groups of 10 male and 10 female rats

were exposed to the same concentrations for 23 days.

All rats survived to the end of the study. Mean body

weights of 1,000 ppm males were significantly less than

those of the chamber controls. The increases in sorbitol

dehydrogenase activities in 500 ppm males and

1,000 ppm males and females were consistent with the

histopathologic evidence of mild hepatotoxi city caused

by 1

- bromopropane. Liver weights of males exposed to

250 ppm or greater and of females exposed to 125 ppm

or greater were significantly increased. Spleen and kid

ney weights of 1,000 ppm females were significantly

increased. E xposure concent ration

- related decreases of

28% in sperm motility and 37% in sperm counts were

seen in the 1,000 ppm group of male rats . Female rats

in all three expos ure groups evaluated exhibited altered

estrous cycle s, spending signifi cantly more time in

extended estrus and less time in extended diestrus

. The

incidences of c ytoplasmic vacuolization of the liver

w ere significantly increased in males exposed to

250 ppm or greater and in female s exposed to 500 ppm or greater. Hepatocyte degeneration was also observ ed

in 1,000 ppm females.



# 3-M ONTH 

# S TUDY IN 

# M ICE

Groups of 10 male and 10 female mice were exposed to

1- bromopropane vapor at concentrations of 0, 62.5, 125,

250, or 500 ppm, 6 hours plus T 90 

### (10 minutes) per day,

5 days per week for 14 weeks. One 250 ppm male and

four males and five females in the 500 ppm groups died

early. Mean body weights of exposed groups were sim

ilar to those of the chamber controls. Lethargy was

observed in males and females exposed to 500 ppm

, and

abnormal breathing was observed in moribund mice

. The kidney, liver, and lung weights of 500 ppm females

were significantly greater than those of the chamber

controls. The kidney weights of 500 ppm males were

significantly decreased. S perm counts in the 500 ppm

group of m ale mice were 28% less than that in the

chamber controls. Female mice exhibited altered

estrous cycles, with females in the 500 ppm group

spending significantly more time in extended diestrus

and those in the 250 ppm group spending signi f icantly

more time in extended estrus compared to the chamber

controls

. Nonneoplastic lesions were observed in the

nose, larynx, trachea, lung, and liver of 500 ppm males

and females and in the adrenal cortex of 500 ppm

females.



# 2-Y EAR 

# S TUDY IN 

# R ATS

Groups of 50 male and 50 female rats were exposed to

1- bromopropane vapor at concentrations of 0, 125, 250,

or 500 ppm, 6 hours plus T 90 

### (10 minutes) per day,

5 days per week for 105 weeks. Survival of 500 ppm

males was significantly less than that of the chamber

control group. Mean body weights of exposed groups

were similar to those of the chamber controls.

Increased incidences of macroscopic, soft, pale

- yellow

to green, variably sized nodules were seen predomi

nantly in the nose and skin of exposed rats. The number

of animals with multiple masses was increased in the

500 ppm groups. In m ost cases, these lesions were

microscopically shown to be suppurative inflammation

,

many with Splendore

- Hoeppli mater ial.

The incidence of adenoma of the large intestine (colon

or rectum ) was significantly greater in 500 ppm females

than in the chamber control group. The incidence of

adenoma of the large intestine in 250 ppm males

exceeded the historical control ranges for inhalation

studies and all routes.

### 1- Bromopropane, NTP TR 564 9

T he incidences of keratoacanthoma, basal cell adenoma,

basal cell carcinoma, or squamous cell carcinoma (com

bined) were significantly greater in all exposed groups

of males than in the chamber control group and

exc eeded the historical control range for inhalation stu dies. The incidences of keratoacanthoma and of kera

toacanthoma or squamous cell carcinoma (combined) in

250 and 500 ppm males were also significantly

increased and exceeded the historical control rang es for

inhalation studies. In 500 ppm females, the incidence of

squamous cell papilloma, keratoacanthoma, basal cell

adenoma, or basal cell carcinoma (combined) exceeded

the historical control range for inhalation studies.

The incidence of malignant mesothelioma was signifi

cantly greater in 500 ppm males than in the chamber

control group.

The incidences of pancreatic islet adenoma in all

exposed groups of males and of pancreatic islet ade

noma or carcinoma (combined) in 12 5 and 250 ppm

males were significantly increased.

Treatment

- related nonneoplastic lesions were observed

in the respiratory system of exposed male and female

rats. In the nose, the incidences of suppurative chronic

inflam mation, chronic active inflammatio n, glandular

hyperplasia, respiratory epithelial hyperplasia (females)

, and respiratory metaplasia of the olfactory epithelium

(females) were increased in all exposed groups . In the

larynx, the incidences of chronic active inflammation

and squamous metapl asia (except 125 ppm females)

were increased in all exposed groups, and the incidences

of suppurative chronic inflammation were increased in

the 500 ppm groups. Also, chronic inflammation of the

lung was observed in the 500 ppm females. In the

trachea, there were increased incidences of chronic

active inflammation in all exposed groups of females

and 500 ppm males

, and the incidence of epithelial

hyperplasia was increased in 500 ppm females.



# 2-Y EAR 

# S TUDY IN 

# M ICE

Groups of 50 male and 50 female mice were exposed to

1- bromopropane vapor at concentrations of 0, 62.5, 125,

or 250 ppm, 6 hours plus T 90 

### (10 minutes) per day,

5 days per week for 105 weeks. Survival of exposed

groups was similar to that of the chamber controls.

Mean body weights of all exposed groups were similar

to those of the chamber controls throughout the study.

In the females, there were increased incidences of alve

olar/bronchiolar adenoma, alveolar/bronchiolar carci

noma, and alveolar/bronchiolar adenoma or carcinoma

(combined); the incidences of alveolar/bronchiolar

adenoma or carcinoma (combined) were significantly

increased in all exposed groups of females. There were

signific antly increased incidences of cytoplasmic vacu

oli zation of the bronchiolar epithelium in all exposed

male groups and regeneration of the bronchiolar epithe

lium in all exposed groups of males and females

.

In the nose, there were significantly increased inci

dences of cytoplasmic vacuolization of the respiratory

epithelium in all exposed groups of males and in 125

and 250 ppm females. There were significantly

increased incidences of respiratory epithelial hyper

plasia in all exposed female groups and in 62.5 and

250 ppm males. There were significantly increased

incidences of respiratory metaplasia of olfactory epithe

lium in 62.5 and 125 ppm males and 125 and 250 ppm

females.

T here were significantly increased incidences of cyto

plasmic vacuolization of respiratory epithelium in the

larynx and trachea of all exposed male groups and in the

trachea of 62.5 and 125 ppm females.



# G ENETIC 

# T OXICOLOGY

1- Bromopropane was not mutagenic in either of two

independent bacterial mutagenicity assays, each con

ducted wit h and without induced rat liver activation

enzymes. Bacterial strains tested included S almonella

typhimurium strains TA97, TA98, TA100, and TA1535,

and Escherichia coli strain WP2 uvrA / pKM101. In

addition, no increases in the frequencies of micronu

cleated normochromatic erythrocytes were seen in male

or female B6C3F1 mice exposed for 3 months to 62.5 to

500 ppm 1

- bromopropane via inhalation.



# C ONCLUSIONS

Under the conditions of these 2

- year inhalation studies,

there was some evidence of carcinogenic activity * of

1- bromopropane in male F344/N rats based on the

occurrence of rare adenomas of the large intestine and

increased incidences of epithelial neoplasms of the skin

(keratoacanthoma, squamous cell carcinoma, and basal

cell neoplasms). Increased incidences of malignant

mesothelioma and pancreatic islet adenoma and

carcinoma (combined) may also have been related to

1- bromopropane exposure. There was clear evidence of

carcinogenic activity of 1

- bromopropane in female

F344/N rats based on increased in cidences of adenoma

of the large intestine. Increased incidences of skin

### 10 1- Bromopropane , NTP TR 564

neoplasms may also have been related to 1

- bromopro

pane exposure. There was no evidence of carcinogenic

activity of 1

- bromopropane in male B6C3F1 mice

exposed to concentrations of 6 2.5, 125, or 250 ppm

1- bromopropane. There was clear evidence of car

cinogenic activity of 1

- bromopropane in female

B6C3F1 mice based on increased incidences of alveo

lar/bronchiolar neoplasms

. Exposure to 1

- bromopropane resulted in increased

incidences of nonneoplastic lesions in the nose of rats

and mice, the larynx of rats and male mice, and the

trachea and lung of female rats and male and female

mice. Suppurative inflammatory lesions with

Splendore

- Hoeppl i material were present primarily in

the nose and skin of male and female rats exposed to

1- bromopropane.

* Explanation of Levels of Evidence of Carcinogenic Activity is on page 13. A summary of the Technical Reports Review Subcommittee

comments and the public discussion on this Technical Report appears on page 1 5.

### 1- Bromopropane, NTP TR 564 11

Summary of the 2

- Year Carcinogenesis and Genetic Toxicology Studies of 1- Bromopropane

Male

F344/N Rats Female

F344/N Rats Male

B6C3F1 Mice Female

B6C3F1 Mice

Concentrations in air 0, 125, 250, or 500 ppm 0, 125, 250, or 500 ppm 0, 62.5, 125, or 250 ppm 0, 62.5, 125, or 250 ppm

Body weights Exposed groups similar to

the chamber control group Exposed groups similar to

the chamber control group Exposed groups similar to

the chamber control group Exposed groups similar to

the chamber control group

Survival rates 23 /50, 26 /50, 18 / 50 , 1 3 /50 3 4/ 50

, 33 /50, 30 / 50

, 2 4/50 37 /50, 33 /50, 32 /50, 36 /50 36 / 50

, 40 / 50 , 3 7 / 50

, 4 2/50

Nonneoplastic effects Nose : inflammation,

suppurative, chronic

(0/50, 1/ 48

, 2 /4 8

, 7 /50) ;

inflammation, chronic

active (29/50, 33/48,

34/48, 35/50); glands,

hyperplasia (5/50, 14/48,

14/48, 15/50)

Larynx : inflammation,

chronic active (21/50,

28/50, 31/50, 26/50) Nose : inflammation,

suppurative, chronic

(0/50, 1/50, 3/49, 7/50);

inflammation, chronic

active (24/50, 37/50,

37/49, 36/50); glands,

hyperplasia (6/50, 23/50,

28/49, 30/50); respiratory

epithelium, hyperplasia

(5/50, 13/50, 9/49, 18/50);

olfactory epithelium,

metaplasia, respiratory

(3/50, 4/50, 6/49, 9/50)

Larynx : inflammation,

chronic active (18/50,

25/50, 30/50, 32/50);

metaplasia, squamous

(3/50, 2/50, 6/50, 21/50)

Lung : inflammation,

suppurative, chroni c

(0/50, 0/50, 0/50, 4/50)

Trachea : inflammation,

chronic active (0/50, 4/50,

1/50, 6/50); epithelium,

hyperplasia (0/50, 0/50,

0/50, 4/50) Lung : bronchiole,

vacuolization cytoplasmic

(0/50, 18 /50, 19 / 49

, 17 / 49 ) ; bronchiole,

regeneration (1/50, 44/50,

38/49, 47/49)

Nose : respiratory

epithelium, vacuolization

cytoplasmic (0/50, 12/50,

19/50, 20/50); respiratory

epithelium, hyperplasia

(16/50, 29/50, 23/50,

26/50) ; olfactory

epithelium, metaplasia,

respiratory (0/50, 7/50,

6/50, 3/50)

Larynx : vacuoliz ation

cytoplasmic (0/48, 5/50,

10/48, 11/50)

Trachea : vacuolization

cytoplasmic (0/49, 15/50,

24/47, 24/50) Lung : bronchiole,

regeneration ( 0/50, 45/50,

43/50, 49/50)

Nose : respiratory

epithelium, vacuolization

cytoplasmic (0/50, 3/50,

5/50, 8/50); re spiratory

epithelium, hyperplasia

(11/50, 25/50, 28/50,

27/50); olfactory

epithelium, metaplasia,

respiratory (0/50, 4/50,

5/50, 14/50)

Trachea : vacuolization

cytoplasmic (0/50, 8/49,

7/50, 4/50)

Neoplastic effects Large intestine : adenoma

(0/50, 0 /50, 2/ 50

, 1 /50)

Skin : keratoacanthoma

(0/50, 3 /50, 6 /50, 6 /50);

keratoacanthoma or

squamous cell carcinoma

(1 /50, 4 /50, 6 /50, 8/50) ;

keratoacanthoma, basal

cell adenoma, basal cell

carcinoma, or squamous

cell carcinoma (1/50,

7/50, 9/50, 10/50) Large intestine None : adenoma

(0/50, 1/50, 2/50, 5/50) Lung :

alveolar/bronchiolar

adenoma (1/50, 6/50,

4/50, 10/50);

alveolar/bronchiolar

carcinoma (0/50, 7/50,

5/50, 4/50);

alveolar/bronchiolar

adenoma or carcinoma

(1/50, 9/50, 8/50, 14/50)

Equivocal findings Malignant mesothelioma :

(0/50, 2/50, 2/50, 4/50)

Pancreatic islets :

adenoma (0/50, 5 /50,

4 /50, 5 /50) ; adenoma or

carcinoma ( 3/50, 10/50,

9/50, 8/50) Skin None : squamous cell

papilloma,

keratoacanthoma, basal

cell adenoma, or basal

cell carcinoma (1/50,

1/50, 1/50, 4/50) None

Level of evidence of

carcinogenic activity Some evidence Clear evidence No evidence Clear evidence

### 12 1- Bromopropane , NTP TR 564

Summary of the 2

- Year Carcinogenesis and Genetic Toxicology Studies of 1- Bromopropane

Genetic toxicology

Bacterial gene mutations: Negative in Salmonella typhimurium strains TA9 7 , TA 98 , TA 100 , and TA153 5 with

and without S9; and in Escherichia coli WP 2 uvrA /pKM101 with and without S9

Micronucleated erythrocytes

Mouse peripheral blood in vivo: Negative

### 1- Bromopropane, NTP TR 564 13



## EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY

The National Toxicology Program describes the results of individual experiments on a chemical agent and notes the strength of the evidence for

conclusions regarding each study. Negative results, in which the study animals do not have a greater incidence of neoplasia than control animals,

do not necessarily mean that a chemical is not a carcinogen, inasmuch as the experiments are conducted under a limited set of conditions

. Positive results demonstrate that a chemical is carcinogenic for laboratory animals under the conditions of the study and ind icate that exposure to

the chemical has the potential for hazard to humans. Other organizations, such as the International Agen cy for Research on Cancer, assign a

strength of evidence for conclusions based on an examination of all available evidence, including animal studies such as those conducted by the

NTP, epidemiologic studies, and estimates of exposure. Thus, the actual det ermination of risk to humans from chemicals found to be

carcinogenic in laboratory animals requires a wider analysis that extends beyond the purview of these studies.

Five categories of evidence of carcinogenic activity are used in the Technical Report series to summarize the strength of evidence observed in

each experiment: two categories for positive results (clear evidence and some evidence) ; one category for uncertain findings (equivocal

evidence) ; one category for no observable effects (no evidence) ; and one category for experiments that cannot be evaluated because of major

flaws (inadequate study) . These categories of interpretative conclusions were first adopted in June 1983 and then revised on March 1986 for use

in the Technical Report series to incorporate more specifically the concept of actual weight of evidence of carcinogenic activity. For each

separate experiment (male rats, female rats, male mice, female mice), one of the following five categories is selected to describe the findings.

The se categories refer to the strength of the experimental evidence and not to potency or mechanism.

**•** Clear evidence of carcinogenic activity is demonstrated by studies that are interpreted as showing a dose

- related (i) increase of

malignant neoplasms, (ii) i ncrease of a combination of malignant and benign neoplasms, or (iii) marked increase of benign neoplasms

if there is an indication from this or other studies of the ability of such tumors to progress to malignancy.

**•** Some evidence of carcinogenic activity is demonstrated by studies that are interpreted as showing a chemical

- related increased

incidence of neoplasms (malignant, benign, or combined) in which the strength of the response is less than that required for clear

evidence.

**•** Equivocal evidence of carcino genic activity is demonstrated by studies that are interpreted as showing a marginal increase of

neoplasms that may be chemical related.

**•** No evidence of carcinogenic activity is demonstrated by studies that are interpreted as showing no chemical

- related increases in

malignant or benign neoplasms

**•** Inadequate study of carcinogenic activity is demonstrated by studies that, because of major qualitative or quantitative limitations,

cannot be interpreted as valid for showing either the presence or absence of carcin ogenic activity.

For studies showing multiple chemical

- related neoplastic effects that if considered individually would be assigned to different levels of evidence

categories, the following convention has been adopted to convey completely the study result s. In a study with clear evidence of carcinogenic

activity at some tissue sites, other responses that alone might be deemed some evidence are indicated as “were also related” to chemical exposure.

In studies with clear or some evidence of carcinogenic activity, other responses that alone might be termed equivocal evidence are indicated as

“may have been” related to chemical exposure.

When a conclusion statement for a particular experiment is selected, consideration must be given to key factors that would extend the actual

boundary of an individual category of evidence. Such consideration should allow for incorporation of scientific experience a nd current

understanding of long

- term carcinogenesis studies in laboratory animals, especially for those evaluat ions that may be on the borderline between

two adjacent levels. These considerations should include:

**•** adequacy of the experimental design and conduct;

**•** occurrence of common versus uncommon neoplasia;

**•** progression (or lack thereof) from benign to malignant n eoplasia as well as from preneoplastic to neoplastic lesions;

**•** some benign neoplasms have the capacity to regress but other s (of the same morphologic type) progress. At present, it is impossible

to identify the difference. Therefore, where progression is known to be a possibility, the most prudent course i s to assume that benign

neoplasms of those types have the potential to become malignant;

**•** combining benign and malignant tumor incidence known or thought to represent stages of progression in the same orga n or tissue;

**•** latency in tumor induction;

**•** multiplicity in site

- specific neoplasia;

**•** metastases;

**•** supporting information from proliferative lesions (hyperplasia) in the same site of neoplasia or other experiments (same lesion in

another sex or species);

**•** presence or absence of dose relationships;

**•** statistical significance of the observed tumor increase;

**•** concurrent control tumor incidence as well as the historical control rate and variability for a specific neoplasm;

**•** survival

- adjusted analyses and false posi tive or false negative concerns;

**•** structure

- activity correlations; and

**•** in some cases, genetic toxicology

.

### 14 1- Bromopropane , NTP TR 564



## NATIONAL TOXICOLOGY PROGRAM BOARD OF SCIENTIFIC COUNSELORS

TECHNICAL REPORTS REVIEW SUBCOMMITTEE

The members of the Technical Reports Revi ew Subcommittee who evaluated the draft NTP Technical Report on 1- bromopropane on

November 19, 2009

, are listed below. Subcommittee members serve as independent scientists, not as representatives of any institution, company,

or governmental agency. In this capacity, subcommittee members have five major responsibilities in reviewing the NTP studies :

**•** to ascertain that all relevant literature data have been adequately cited and interpreted,

**•** to determine if the design and conditions of the NTP studies were appropriate,

**•** to ensure that the Technical Report presents the experimental results and conclusions fu lly and clearly,

**•** to judge the significance of the experimental results by scientific criteria, and

**•** to assess the evaluation of the evidence of carcinogenic activity and other observed toxic responses.



### Raymond F. Novak, Ph.D., Chairperson

Children’s Hospital of Michigan

Wayne State University School of Medicine

Detroit, MI



### Tracie E. Bunton, D.V.M., Ph.D.

Toxicology Consultant

Eicarte LLC

Gettysburg, PA



### Russell C. Cattley, V.M.D., Ph.D.

Amgen

Thousand Oaks, CA



### David A. Eastmond, Ph.D.

, Principal R eviewer

Department of Cell Biology and Neuroscience

University of California

Riverside, CA



### Stephen W. Looney, Ph.D.

Department of Biostatistics

Medical College of Georgia

Augusta, GA



### Mitzi Nagarkatti, Ph.D.

Department of Pathology, Microbiology, and Immu nology

University of South Carolina School of Medicine

Columbia, SC 

### Michael V. Pino, D.V.M., Ph.D.

, Principal Reviewer

Drug Safety Evaluation

Sanofi

- aventis

Alfortville, France



### Ken neth M. Portier, Ph.D.

, Principal Reviewer

American Cancer Society

Atlanta, GA



### Jim E. Riviere, D.V.M., Ph.D.

College of Veterinary Medicine

North Carolina State University

Raleigh, NC



### James L. Sherley, M.D., Ph.D.

Programs in Regenerative Biology and Cancer

Boston Biomedical Research Institute

Watertown, MA



### Justin G. Teeguarden, Ph.D.

Pacific Northwest National Laboratory

Richland, WA

### 1- Bromopropane, NTP TR 564 15



## SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMITTEE COMMENTS

On November 19, 2009, the draft Technical Report on the

toxicology and carcinogen esis studies of 1

- bromopropane

received public review by the National Toxicology

Program’s Board of Scientific Counselors’ Technical

Reports Review Subcommittee. The review meeting was

held at the National Institute of Environmental Health

Sciences, Research Triangle Park, NC

.

Dr. D .L. Morgan, NIEHS, introduced the toxicology and

carcinogen esis studies of 1

- bromopropane by reviewing

the chemical’s extensive use as a solvent, the rationale for

and design of the inhalation studies, the observed toxicity

and body weight effec ts in the short

- term studies, and the

observed neoplasms and nonneoplastic lesions in the

long

- term studies. The proposed conclusions were some

evidence of carc i nogenic activity of 1

- bromopropane in

male rats, clear evidence of carcinogenic activity of

1- bromoprop a ne in female rats, no evidence of carcino

genic activity of 1

- bromopropane in male mice, and clear

evidence of carcinogenic activity of 1

- bromopropane in

female mice.

Dr. Pino, the first principal reviewer, had no scientific

criticisms. He sugg ested that basal cell carcinomas and

adenomas be listed in the results table for rats. He also

suggested that pancreatic carcinomas and adenomas be

added to the conclusion statement for male rats. He

asked for clarification of the time and extent of the

Splendore

- Hoepp l i inflammatory lesions and suggested

specifying the types of skin tumors in rats. He also ask ed

if some decreased tumor incidences should be discussed

as in some other reports

.

Dr. Morgan agreed to include mention of the basal cell

carcin omas in rat skin in the results section. He noted

that the pancreatic carcinomas had not been included

because they did not increase with dose and were not

considered treatment related. Dr. Morgan said the

decreased incidences of skin sarcomas and liver ade

nomas and carcinomas in female rats could be included

under other findings in the resul t s section.

Dr. Eastmond, the second principal reviewer, felt the

study was well conducted and the report well written. He

felt the proposed conclusion of some evidence for the intestinal tumors was fair even without statistical sig

nifi cance, given the rarity of those tumors. He suggested

some caveats in reporting the results of the mutagenicity

tests for this volatile chemical. He asked for clarification

of t he T 90 

### period at the start of the animal exposure period

in the inhalation chambers and inquired if the pathology

slides were coded during the initial diagnoses.

Dr. Morgan said more description would be provided

about the mutagenicity test methodology an d the chamber

startup periods. Dr. M.F. Cesta, NIEHS, said slides are

not coded during the initial readings, though they are in

subsequent reviews.

Dr. Portier, the third principal reviewer, agreed with the

proposed conclusions.

Dr. Cattley expressed a preference for specifying the skin

neoplasm types in the conclusion statement.

Dr. Nagarkatti asked if any further studies were per

formed to determine if the chemical produced any sensi

tization leading to development of the Splendore

- Hoeppli

bodies. Dr

. Morgan said that while no other studies were

performed, a number of other brominated compounds

were immunosuppressive.

Dr. D. Lynch, NIOSH, commented that the present data

would be helpful in the current development of exposure

guidelines.

Dr. Pino moved that the conclusion in male rats specify

adenomas and carcinomas (combined) for pancreatic

lesions, and that the skin tumors be identified as epithelial

tumors. Dr. Cattley added that the types of skin lesions

should be specified, and Dr. Pino ident ified keratoac

antho m as, squamous cell carcinoma, or basal cell neo

plasms to be added parenthetically as the skin neoplasms

for male rats.

Dr. Portier moved and Dr. Nagarkatti seconded that the

conclusions be accepted as modified. Dr. Teeguarden

was recused from the peer review and vote because of a

conflict of interest and the motion was approved unani

mously with nine votes.

### 16 1- Bromopropane , NTP TR 564

### 17



# INTRODUC TION

1- BROMOPROPANE

CAS No. 106

- 94 -5

Chemical Formula: C 3 

### H 7 

### Br Molecular Weight: 12 2. 99

Synonyms : 1- BP; p ropyl bromide; n- BP; N- propyl bromide



# C HEMICAL AND 

# P HYSICAL 

# P ROPERTIES

1- Bromopropane is a colorless to pale yellow liquid

with a strong

, characteristic odor. The boiling point is

71 ° C and the vapor pressure is 110.8 mm Hg a t 20 ° C

(UNEP, 2001; Merck , 2006) . 1 -B romopropane is less

flammable than many other halogenated alkanes at

room temperature. Thermal decomposition of 1

- bromo

propane produces hydrogen bromide. 1

- Bromopropane

reacts with oxidizing agents to produce hazardous

flammable compounds and can react with water to form

acids. The 1

- bromopropane used for industrial applica

tions and in most commercial products is stabilized to

inhibit hydrolysis, and formulated to improve perfor

mance and enhance the useable life of the product

( UNEP, 2001; HSDB

, 200 9).



# P RODUCTION



# , U SE



# ,

AND 

# H UMAN 

# E XPOSURE

Industrial production of 1

- bromopropane generally

involves reacting propanol with an excess of hydr ogen

bromide gas. The principal product is 1

- bromopropane with small amounts of 2

- bromopropane (isopropyl bro

mide) and other by

- products. Various modifications of

the synthetic method are used to increase the purity of

the 1

- bromopropane, and distilla tion procedures are

often used to remove most of the by

- products (UNEP

, 2001)

.

In 2006, worldwide annual production capacity of

1- bromopropane was estimated at greater than

20,000 metric tons (44 million pounds/year), of which

5,000 metric tons were thought to be used as a pharma

ceutical intermediate or process agent. U nited S tates

produ ction was estimated at approximately 5,000 metric

tons (11 million pounds/year) and growing at a rate of

15 % to 20% per year (UNEP

, 2006)

.

In the early to mid 1990s, 1

- bromopropane was used

primarily as an intermediate in the production of pesti

cides, quaternary ammonium compounds, flavors and

fragrances, pharmaceuticals, and other chemicals in

well

- controlled, closed processes ( Hanley et al ., 2006 )

. In the mid to late 1990s, 1

- bromopropane was intro

duced as a less toxic replacement for methylene chloride

### 18 1- Bromopropane , NTP TR 564

in emissive app lications such as vapor and immersion

degreasing operations and critical cleaning of electron

ics and metals. 1

- Bromopropane has a relatively high

vapor pressure and is well suited for use in cleaning

equipment where it is repeatedly vaporized. Because i t

is relatively nonflammable , 1- bromopropane can be

used safely in metal cleaning processes where heating is

required. 1- Bromopropane was also introduced as a

nonflammable, nontoxic, fast

- drying, and inexpensive

solvent for adhesive resins. Aerosol

- appli ed adhesives

containing 1

- bromopropane were used extensively by

foam fabricating companies (Hanley et al ., 2006)

. Because 1

- bromopropane has a relatively short atmo

spheric ha lf

- life (16 days)

, it is considered to have a

relatively low ozone depletion potential and has been

marketed as a replacement for ozone depleting refriger

ants (e.g.

, chlorofluorohydrocarbons and hydrochloro

fluorocarbons) ( Fed

. Regist

. , 2000)

.

Emissive use of 1

- bromopropane leads to substantial

dermal and inhalation exposure of workers. Currently

, there are no data available for dermal exposure to

1- bromopropane. T he N ational I nstitute for O ccupa

tional S afety and H ealth (NIOSH) obtained personal

breathing zone exposure data for workers at three foam

fabrication plants that used spray ad hesive s containing

1- bromopropane. In these three plants, 8 -h our time

weighted average ( TWA ) exposures ranged from 18 to

381 ppm (mean = 142 ppm) (NIOSH

, 2002

, 2003)

. In

two of these plants the 8 -h our TWA exposure concen

trations of 1

- bromopropane were significantly lowered

(1.2 to 58 ppm; mean = 19 ppm) after improvements

were made in ventilation ( NIOSH, 200 2)

.



# A BSORPTION



# , D ISTRIBUTION



# , M ETABOLISM



# , AND 

# E XCRETION



## Experimental Animals

Rats (strain not specified) dosed intraperitoneally with

[ 14 

### C] -1- bromopropane exhaled unchanged 1

- bromopro

pane and [ 14 

### C]- carbon dioxide (Jones and Walsh, 1979)

. The metabolites

, N

- acetyl

- S- (2

- hydroxypropyl)cysteine,

N

- acetyl

- S

- propylcysteine, and the cor responding

S

- oxide [ N

- acetyl -3- (propylsulfinyl)alanine] were iden

tified in the urine (Barnsley et al ., 1964 , 1966 ; Barnsley,

1966; Jones and Walsh, 1979). Lee et al. (2007 a )

observed S

- propyl glutathione in livers of BALB/ c mice

following a single oral dose of 1

, 000 mg/kg 1

- bromo

propane, with m aximum levels of S

- propyl glutathione

detected 6 h ours after administration

. Exposure to

1- bromopropane has been shown to de plete glutathione

in rat brain in a dose

- dependent manner, independent of

duration of exposure (Wang et al ., 2003)

. In male Wistar rats exp osed to 700 or 1

, 500 ppm 1- bromopro

pane vapor

, 6 h ours a day, 5 days a week, for 3 or more

weeks

, 1- bromopropane in blood decreased rapidly to

the detection limit within 0.7 h ours after exposure

ceased. On the other hand, b romi d e ion s persisted

longer in both blood and urine; the biological half

- life

of bromi d e ion was 4.7 to 15.0 days in blood and 5.0 to

7.5 days in urine (Ishidao et al

., 2002)

.

T he N ational Toxicology Program (NTP) examined t he

disposition and metabolism of 1

- bromopropane and the

factors influencing them in male F344 /N rats and

B6C3F1 mice following inhalation exposure (800 ppm)

or intraveno us administration (5, 20, or 100 mg/kg)

(Garner et al ., 2006). [ 1,2,3

- 13 

### C] -1- bromopropane and

[ 14 

### C] -1- bromopropane were co

- administered to enable

characterization of urinary metabolites using nuclear

magnetic resonance spectroscopy and liquid chroma

tography coupled with either tandem mass spectrometry

or radiochromatography. By 4 hours f ollowing intra

venous administration, rats and mice exhaled a majority

of the administered [ 14 

### C] -1- bromopropane dose as either

volatile organic compound s (rats, 50 % to 71%; mice,

39 % to 48%) or carbon dioxide (rats, 10 % to 30%; mice,

19 % to 26%). The radioactivity was also excreted in

urine (rats, 13 % to 19%; mice, 13 % to 23%) and feces

(rats, < 2 %; mice, 4%) or retained in tissues and carcass

(rats, ≤ 6%; mice, < 4%). N

- acetyl

- S

- propyl

- cysteine,

N

- acetyl

- S

- (2

- hydroxypropyl)cysteine, N

- acetyl -3- (pro

pylsulfinyl)alanine, 1

- bromo -2- hydroxypropane

- O

- glu

curonide, N

- acetyl

- S

- (2

- oxopropyl)cysteine, and N

- ace

tyl -3- [(2

- oxopropyl)sulfinyl]alanine were the urinary

metabolites characterized in rats and mice following both

inhalation exposure and intravenous administration

.

In rats, but not in mice, the route of elimination and the

metabolite distribution changed significantly as the dose

increased

, with t he percentage of dose excreted as vola

tile organic compound s increasing significantly between

the mid

- and high

- dose groups (Garner et al ., 2006)

. Concomitantly, the percentage of the dose exhaled as

carbon dioxide decreased. An investigation of the

molar ratio o f exhaled carbon dioxide to the total

released bromide showed that the proportion of

1- bromopropane metabolized via oxidation to pathways

dependent on glutathione conjugation with bromine

replacement decreased with increasing dose in rats. A s

the dose inc reased, the relative proportion of N

- acetyl

S

- propylcysteine and N

- acetyl

- S

- (2

- hydroxypropyl)

- cysteine shifted such that the N

- acetyl

- S

- propylcysteine

peak predominated at 100 mg/kg accounting for more

than 80% of the urinary radioactivity and indicating

saturation of oxidation pathways. Rats pretreated with

1- aminobenzotriazole, a potent inhibitor of cytochrome

### 1- Bromopropane, NTP TR 564 19

P450

, had decreased total radioactivity excreted in

urine, exhaled as carbon dioxide , or retained in liver

with a concomitant increase in radioactivity in expired

volatile organic compound s

. The number of urinary

metabolites was reduced from 10 to one with N

- acetyl

S

- propylcysteine accounting for more than 90% of the

total urinary radioactivity in the pretreated rats

. These

data demonstrate a role for P450 and glutathione in the

dose

- dependent metabolism and disposition of 1

- bromo

propane in the rat

.

Garner et al. (200 7 ) also investigated the contribution of

cytochrome P450 2E1 ( CYP2E1 ) to 1- bromopropane

metabolism using CYP2E1 knockout and wild type

mice. T he mercapturic acid of 1

- bromo -2- hydroxy

propane

, N

- acetyl

- S

- (2

- hydroxypropyl)cysteine, was

observed as the major urinary metabolite in wild type

mice; the products of direct conjugation of 1

- bromo

propane with glutathione were reported ly insignificant.

The ratio of glu tathione conjugation to 2

- hydroxylation

was increased five fold in the CYP2E1 knockout mice

relative to wild type mice. In Sprague

- Dawley rats

exposed to as much as 1, 800 ppm 1

- bromopropane / kg

(6 h ours a day, 5 d ays a week, for 8 weeks), Kim et al.

(1999) showed that 1

- bromopropane primarily induce d

CYP2E1 as the major form of CYP and that glutathione

S

- transferase (GST) enzymes played important roles in

the metabolism. Sex difference in the metabolic

mechanism in the rat liver was also proposed (Kim

et al ., 1999)

. These results suggest that CYP2E1 is a

major metabolic enzyme of 1

- bromopropane al though

other P450s may be involved. Based on these findings,

Garner et al . (2006) proposed a scheme for the metab

olism of 1- bromopropane in rodents (Figure 1).



## Humans

N

- acetyl

- S

- propylcysteine was detected in the urine of

occupationally exposed workers (Ichihara et al ., 2004 a).

In another study, workers exposed to 1

- bromopropane at

two facilities using 1

- bromopropane adhesives showed a

significant association of 48 -h our urinary bromi de ion

concentration with 1- bromopropane exposure measured

in the breathing zone (Hanley et al ., 2006)

. These data

demonstrate that urinary elimination is an important

excretion pathway of 1

- bromopropane in humans. 

# G ENERAL 

# T OXICITY



## Experimental Animals

The acute toxicity of 1

- bromopropane has been inves

ti gated in rats by several routes of administration. The

inhalation LC

50 

### for 1

- bromopropa ne was reported as

7,000 ppm for 4 hours in Wistar rats ( ACGIH, 2005 ).

For non

- inhalation routes, the LD

50 

### was reported as

2.9 g/kg in rats and 2.5 g/kg in mice by intraperitoneal

injection ( Patty’s , 2001 ). The LD

50 

### for oral and dermal

routes of administration in rats is greater than

2,000 mg/kg ( ACGIH, 2005 ).

Th e LC

50 

### for 2

- bromopropane, a structurally related

brominated alkane is reported as 31,171 ppm for a

4- hour inhalation exposure in mice ( ACGIH, 2005 ).

The L D

50 

### for intraperitoneal injection of 2

- bromopro

pane in mice is 4 .8 g/kg ( Lewis , 1996 ). In rats, the

LD

50 

### for oral administration of 2

- bromopropane is

greater than 2,000 mg/kg ( ACGIH, 2005 )



# R EPRODUCTIVE

AND 

# D EVELOPMENTAL 

# T OXICITY



## Experimental Animals

1- Br omopropane was shown to cause developmental

toxicity in rat pups whose dams were exposed during

the period of in utero development (Hunting d on Life

Sciences, 2001) . Decreased fetal weights and increased

incidences of skeletal variations were observed in pups

of dams exposed to 1

- bromopropane at 500 ppm or

greater for 6 hours/day during gestation day s 6 t hrough

19.

A two

- generation reproductive toxicity study (WIL

Research Laboratories, 2001) showed exposure of rats

to 250 ppm or greater altered numerous reproductive

endpoints in both females and males. These included

decreased sperm motility and percent normal sperm in

males and decreased litter size, decreased numbers of

implantation sites, increased ovarian follicular cysts,

and increased estrous cycle length in females. Another

study in Wistar rats (Ichihara et al ., 2000a) comparing

reproductive toxicities of 1

- and 2

- bromopropane

### 20 1- Bromopropane , NTP TR 564

F IGURE 

### 1

Proposed Metabolism of 1

- Bromopropane in Rodents

Metabolites shown in bold font were characterized in urine

from rodents exposed to 1

- bromopropane (Garner et al ., 2006)

### 1- Bromopropane, NTP TR 564 21

demonstrated that exposure to 400 or 800 ppm 1

- bromo

propane for 12 weeks (8 hours/day, 7 days per week)

inhibited sperm count and sperm motility ; it was less

toxic than 2

- bromopropane. Yamada et al

. (2003)

rep orted that female Wistar rats exposed to 400 ppm

1- bromopropane 8 hours /day for 12 weeks had a signif

i cant increase in the number of irregular estrous cycles

with extended diestrus. Histological examination of the

ovary showed a significant reduction in the number of

normal antral follicles, and a decrease in the number of

normal growing follicles in the 400 ppm group.



## Humans

The NTP Center for the Evaluation of Risks to Human

Reproduction (CERHR) evaluated the potential for

1- bromopropane to produce adverse reproductive and

developmental effects in humans (NTP, 2003). CER HR

concluded that there was convincing evidence for

reproductive and developmental toxicity in experimen

tal animals. Evidence in humans was limited, but in the

monograph, not e was made of a new case that was not

available to the expert panel indicating p ositive findings

in women (altered menstruation) occupationally

exposed to 1

- bromopropane. The overall NTP conclu

sion was that “there is serious concern for reproductive

and developmental effects of 1

- bromopropane at the

upper end of the human occupation al exposure range

(18 to 381 ppm).” “Serious concern” is the highest

level of NTP conclusion regarding the possibilities that

human development and reproduction might be

adversely affected.



# N EUROTOXICITY



## Experimental Animals

In an animal study designed to evaluate the neurotoxic

effects of 2

- bromopropane on rat tail nerve, 1

- bromo

propane was used as a negative control (Yu et al

., 1998) . Wistar r ats were exposed to air, 100 or

1, 000 ppm 2

- bromopropane or 1

, 000 ppm 1

- bromopro

pane 8 h ours /d ay , 7 d ays pe r week for up to 7 weeks.

Unexpectedly , 1- bromopropane was found to be more

neurotoxic than 2- bromopropane

. Electrophysiological

changes (decreased maximum motor nerve conduction

velocity and increased distal latency) and histopatho

logic changes in peripheral nerves were detected in rats

exposed to 1

, 000 ppm 1

- bromopropane for 4 weeks.

The exposures were discontinued after 5 weeks because

of hindlimb paralysis in exposed rats. No changes were

observed in rats exposed to the same concentration of

2- bromopropane. Histopathologic changes were also

observed in axons in the gracil e nucleus and in Purkin j e

cells in the cerebellum, suggesting that 1

- bromopropane may also be toxic to the CNS. Ohnishi et al

. (1999) also

reported that exposure to 1

, 500 ppm 1

- bromopropane

6 h ours /day, 5 days per w ee k for 4 w ee ks resulted in

degeneration of Purkin j e cells in the rat cerebellum. In

a subsequent inhalation study, male Wistar rats exposed

to 800 ppm 1

- bromopropane showed myelin degenera

tion in the peripheral nerves, pre

- terminal axon swelling

in the gracile nucleus in the medulla oblongata, weight

loss in the cere brum , limb muscle weakness, distal

latency prolongation, and decrease of motor nerve con

duction velocity (Ichihara et al ., 2000 b).



## Humans

The first case of 1

- bromopropane toxicity in humans

was reported by Sclar (1999). A worker developed

weakness in the proximal extremities and right hand

after using an industrial metal cleaning and degreasing

solvent that contained 1

- bromopropane (95.5% by

weight) to strip metal for about 2 months. Nerve con

duction studies indicated a primary symmetric demy

elinating p olyneuropathy, and magnetic resonance

imaging scans revealed evidence of central nervous

system (CNS) involvement. Subsequently, neurological

signs of toxicity were reported in three workers at a

foam fabrication factory where they used a spray adhe

sive containing 1

- bromopropane (Ichihara et al

., 2004b) . The workers complained of numbness in the

feet, t highs, and buttocks, and reduced sensations of

vibration in the feet suggesting peripheral neuropathy.

Other symptoms such as headache, dizziness, and mem

ory loss suggested adverse effects on the CNS. NIOSH

reported that four additional workers in a dif ferent

cushion manufacturing company were hospitalized

about a year earlier with similar neurological symptoms

(NIOSH, 2002) . All four workers had elevated serum

bromide levels; however, exposure

- related effects on

blood cell counts, male reproductive functions, or nerve

conduction vel ocity were not detected. NIOSH con

ducted health hazard evaluations at another foam

cushion manufacturing plant where 1

- bromopropane

was used in the spray adhesive (NIOSH, 2003)

. Workers complained of headaches, painful tingling in

the hands or feet, tremor, and feeling drunk when not

drinking, indicating possible CNS and peripheral

neurotoxicity.

In 1995, a new cleaning solvent containing about 97%

2- bromopropane (isopropyl bromide) was introduced in

a Korean electronics manufacturing company to replace

Freon (CFC

- 113). Workers exposed to 2

- bromopropane

presented with symptoms such as hand numbness, sug

gesting that 2

- bromopropane exposure caused polyneu

ropathy (Kim et al ., 1996; Park et al ., 1997)

.

### 22 1- Bromopropane , NTP TR 564



# I MMUNOTOXICITY

1- Bromopropane was demonstrated to cause immuno

suppression after oral administration to mice. A single

treatment of female BALB/c mice with 1

- bromopropane

by gavage significantly suppressed the antibody

resp onse to T

- cell dependent antigen and the production

of splenic intracellular IL

- 2 in response to conconavalin

A ( Lee et al ., 2007a ). Similarly, oral treatment with

1,3

- dibromopropane significantly suppressed the anti

body response to T

- dependent antigen i n a dose

- depen

dent manner. Mice treated with 1,3

- dibromopropane for

7 consecutive days suppressed the antibody response to

T- dependent antigen sheep red blood cells ( SRBC ) ( Lee

et al ., 2007b) . The structurally related chemical 2

- bro

mopropane was first indicated to be immunotoxic in a

study designed to evaluate reproductive toxicity in male

rats. In this study, leukocyte numbers were significantly

decreased in rats treated with 2

- bromopropane con

centrations that caused reproductive toxicity ( Yu et al

., 199 7 ). In a subsequent study ( Jeong et al ., 2002)

, 2- bromopropane was shown to cause immunosuppres

sion in rats following oral exposure to 1,000 mg/kg of

2- bromopropane for 28 consecutive days. 2

- Bromopro

pane treatment significantly reduced numbers of leuko

cytes, spleen cells, and thymic cells and suppressed the

antibody response to SRBC s.

As part of the inhalation studies reported in this

Technical Report, t he potential immunotoxicity of

1- bromopropane was investigated because of the earlier

report s of immunosuppression following oral exposure

to 1

- bromopropane (Lee et al ., 2007a) and the immu

notoxicity of structurally related chemicals. In an NTP

study, 1-b romopropane was found to cause some evi

dence of immunosuppression in F 344/N rats and

B6C3F 1 mice after inhalation exposure for 4 and

10 weeks ( Anderson et al ., 20 10 ). Significant decreases

in total spleen cells and T

- cells were detected after

exposure for 4 weeks to 1

- bromopropane concentrations

ranging from 125 to 1,000 ppm. In addition, significant

decreases in the IgM response to SRBCs were observed

in both species after exposure to 1

- bromopropane.



# C ARCINOGENICITY

There have been no carcinogenicity studies of 1

- bromo

propane in experimental animals or epidemiology stu

dies in humans. Ho wever, NTP studies in rodents have

demonstrated that several structurally related bromi

nated hydrocarbons are potent multisite carcinogens.

Inhalation exposure of F344 /N rats to 1,2

- dibromo

ethane (ethylene dibromide) caused carcinomas and

adenocarcinoma s of the nasal cavity and hemangiosar

comas of the circulatory system in male s and females

, mesotheliomas of the tunica vaginalis and adenomatous

polyps of the nasal cavity in males, and fibroadenomas

of the mammary gland and alveolar/bronchiolar adeno

mas and carcinomas in females (NTP, 1982 a)

. 1,2

- Dibromoethane was also carcinogenic in B6C3F1

mice

, causing alveolar/bronchiolar adenomas and carci

nomas in males and females; and hemangiosarcomas of

the circulatory system, fibrosa r comas in subcutaneous

tiss ue, carcinomas of the nasal cavity, and adenocarci

nomas of the mammary gland in females. Inhalation

exposure to 1,2

- dibromo -3- chloropropane caused carci

nomas, squamous cell carcinomas, and adenocarcino

mas of the nasal cavity and squamous cell papilloma s of

the tongue in male and female F344 /N rats (NTP

, 1982b ) . In B6C3F1 mice

, 1,2

- dibromo -3- chloropropane

caused adenocarcinomas of the nasal cavity and papil

lary carcinomas in females and squamous cell carci

nomas of the nasal cavity and alveolar/bronchiolar

adenomas and carcinomas in males and females. In

F344/N rats and B6C3F1 mice, d ermal application of

2,3

- dibromo -1- propanol caused tumors at multiple sites

(NTP , 1993 ).



# G ENETIC 

# T OXICITY

Published data on t he genotoxicity of 1

- bromopropane

suggest that

, when tested appropriately to control for

volatility, the compound demonstrates direct

- acting

mutagenicity in bacteria, and it has a weak potential for

induction of chromosomal damage in vitro and in vivo

in s ome mammalian test systems. Barber et al . (1981)

reported that 1

- bromopropane, tested over a concentra

tion range of 1.1 to 20.3 μ M, was mutagenic in

Salmonella typhimurium strains TA100 and TA1535,

with or without induced rat liver S9 activation enzymes;

the lowest effective concentration was 4.9 μ M, and

potency of response, which directly correlated with con

centration, was unaffected by the addition of S9, thus

indicating that 1

- bromopropane is a direct

- acting muta

gen in Salmonella . Barber et al

. (1981) used a closed

system to control for the volatility of 1

- bromopropane.

Yu et al . (2008) conducted a dominant lethal assay in

male ICR mice administered 1

- bromopropane (300 or

600 mg/kg per day) by gavage for 10 days and mated

once weekly for 6 weeks to undosed female mice.

Females were sacrificed and uterine contents were

examined on gestation day s 15 through 17. No signifi

cant treatment

- related increases in pre

- or post

- implan

tation losses, resorptions, nonviable fetuses, or other

fertility endpoints were seen at either dose at any mating

time point. The authors concluded that under these

experimental conditions, 1

- bromopropane did not

### 1- Bromopropane, NTP TR 564 23

induce dominant lethal mutations in germ cells of male

mice. Negative results were also reported from an ear

l ier dominant lethal study conducted in male Sprague

Dawley rats treated with 400 mg/kg per day 1

- bromo

propane by gavage for 5 consecutive days and mated to

undosed females once weekly for 8 successive weeks

(Saito

- Suzuki et al ., 1982); females were sacrificed 13

to 14 days after mating and uterine contents showed no

significant treatment

- related changes in most indicators

of fertility or embryonic deaths compared with the

vehicle controls. The one exception was a marginally

elevated frequency of dead implants observed at mating

week 8, in the absence of a significant dominant lethal

mutation index. In a parallel investigation at the same

laboratory, male Sprague

- Dawley rats were treated with

200 mg/kg per day 1,2

- dibromopropane for 5 days and

mated to und osed females for 8 weeks (Saito

- Suzuki

et al ., 1982). Examination of uterine contents at gesta

tion day 13 or 14 revealed small increases in dead

implants and in the dominant lethal mutation index in

the week -1 mating group, but no other indications of

adverse effects were observed for 1,2

- dibromopropane.

In contrast, 1,2,3

- tribromopropane (50 mg/kg per day

for 5 days) induced a significant increase in dominant

lethal mutations in male Sprague

- Dawley rats in

po st

- meiotic sperm (primarily early spermatids, mating

weeks 5 and 6) (Saito

- Suzuki et al ., 1982).

NIOSH assessed DNA

- damage levels, measured by the

alkaline single cell gel electrophoresis ( c omet) assay in

peripheral leukocytes of 64 workers from two facil ities

where spray adhesive s containing 1

- bromopropane w ere

used (Toraason et al ., 2006). Workers were divided into

two groups based on 1

- bromopropane exposure level

(high or low); bromide levels in blood were used as an

internal confirmation of exposure, along with personal

breathing zone air samples that were used to calculate

TWA s of 1

- bromopropane concentrations. Although

the high exposure group showed significantly increased

levels of urine and serum bromide compared with the

low exposure group, no si gnificant differences were

seen in the levels of DNA damage between the two

exposure groups. Overall DNA damage measurements

made at the end of the work week in one facility showed

a small but not statistically significant increase, the

result of a signif icant (P<0.05) increase in the low est

exposure group only, while end

- of

- week DNA damage

measures at the second facility were actually lower in

both exposure groups than at the beginning of the work

week. Polymorphisms in GST genes GSTM1 and

GSTT1 were considered in the data analysis process

along with other factors associated with DNA damage

induction such as smoking, age, and gender. In addition to the in vivo assessments of DNA damage in exposed

workers, whole blood cultu res from an unexposed indi

vidual were treated in vitro with either 1

- bromopropane

or 2

- bromopropane, and evaluated for DNA damage

using the c omet assay (Toraason et al ., 2006). Both

1- bromopropane and 2

- bromopropane induced signifi

cant increases in DNA damage at the highest concentra

tion tested, 1 mM; in addition, both compounds

increased the number of apoptotic cells (cells with

diffuse DNA) at 0.1 mM, a 10

- fold lower concentration

than required to induce significant levels of DNA

damage.



# R EGULATORY 

# S T ANDARDS

The ozone

- depleting solvents methyl chloroform and

CFC

- 113 were phased out of production in the United

States beginning in 1996 under the United States

Environmental Protection Agency (USEPA) regulations

and under the Montreal Protocol on Substance s that

Deplete the Ozone Layer. In the late 1990s, 1

- bromo

propane was introduced as a replacement for methyl

chloroform and CFC

- 113 for cleaning metal parts and

electronics and as a nontoxic replacement for methylene

chloride and methyl chloroform adhesi ves

. As the

United States Occupational Safety and Health Adminis

tration ( OSHA ) began phasing in a more restrictive

exposure standard for methylene chloride, the use of

1- bromopropane in adhesives increased. In April 2000,

OSHA ’ s final phase for instituting a workplace expo

sure standard for methylene chloride took full effect

, leading companies to switch to 1

- bromopropane rather

than install ventilation or other controls needed to

reduce employee methylene chloride exposures to the

requisite 25 p pm level. B ecause 1- bromopropane was

not regulated to protect workers, consumers, or the

environment , t he increased production and use was a

concern

.

Several companies petitioned the USEPA ’s Significant

New A lternatives P olicy (SNAP) program to accept the

use of 1

- bromopropane as an alternative to ozone

depleting solvents ( Fed

. Regist

. , 1999) . In 2007, the

US EPA issued a final ruling, finding 1

- bromopropane

acceptable as a substitute for methyl chloroform and for

CFC

- 113 in metals, electronics and precision cleaning

( Fed

. Regist

. , 2007a) . This rule did not address the use

of 1

- bromopropane as an aerosol solvent or as a carrier

solvent in adhesives or coatings. The US EPA proposed

a new rule to prohibit the use of 1

- bromopropane as a

carrier solvent in adhesives and as a solvent in aerosol

cans, and restrict the use of 1

- bromopropane to that of a

carrier solvent in coatings ( Fed

. Regist

. , 2007b)

. The

### 24 1- Bromopropane , NTP TR 564

American Conference of Governmental Industrial

Hygienists (2009) has recommended a threshold limit

value 8

- hour time

- weighted average for 1

- bromopro

pane of 10 ppm.



# S TUDY 

# R ATIONALE

1- Bromopropane was nominated by OSHA for evalua

tion of toxicokinetics, genetic toxicity, reproductive and

developmental toxicity, neurotoxicity, and carcinoge

nicity . The nomination was based on the potential for

widespread occupational and en vironmental exposure to

1- bromopropane and a lack of toxicity and carcinoge

nicity data. Because previous short

- term studies dem

onstrated that 1

- bromopropane is a neurotoxicant and a

reproductive toxicant in animals (Yu et al ., 1998;

Ichihara et al , 2000a), the main focus of the current

studies was to evaluate the chronic toxicity and carcino

gen icity of 1

- bromopropane

.

The 2

- week, 3

- month, and 2

- year studies were con

ducted in male and female F344/N rats and B6C3F1

mice to evaluate the toxicity an d carcinogenicity of

1- bromopropane in association with inhalation expo

sure. Additional studies were conducted to evaluate the

absorption, distribution, metabolism, and excretion of

1- bromopropane and its potential immunotoxicity

(Garner et al ., 2006; An derson et al ., 20 10 ).

### 25



# MATERIALS AND METHOD S

P ROCUREMENT

AND 

# C HARACTERIZATION

OF 

# 1-B ROMOPROPANE

1- Bromopropane was obtained in 55

- gallon metal drums

from Diaz Chemical Corporation ( Holley

, NY ) in on e

lot (106

- 015) and from Albemarle PCC (Thann, France)

in one lot ( 1581313004 ). Lot 106

- 015 was used in the

2- week and 3

- month studies, and lot 1581313004 was

used during the 2

- year st udie s. Identity and purity

analyses were conducted by the study laboratory a t

Battelle Toxicology Northwest (Richland, WA), and by

Chemir/Polytech Laboratories, Inc. (Maryland Heights,

MO) (Appendix I)

. Reports on analyses performed in

support of the 1- bromopropane studies are on file at the

National Institute of Environmental Health Sciences

.

Both lots of the chemical, a colorless to pale yellow

liquid with a strong

, characteristic odor, were identified

as 1- bromopropane by Chemir/Polytech Laboratories

, Inc., by infrared and 1 

### H- nuclear magnetic resonance

spectroscopy.

For lot 106

- 015, Karl Fischer titration indicated a water

content of 39 ppm. Elemental analyses for carbon,

hydrogen, nitrogen, and sulfur were in agreement with

the theoretical values for 1

- bromopropane; residual HBr

was determined to be 1.3 ppm. Gas chromatography

(GC) indicated one major peak and three impurities with

areas exceeding 0.1% of the total peak area. Using

prepared standards, the i mpurities were identified as

1- propanol (0.14%), 2

- bromopropane (0.11%), and

di

- n

- propyl ether (0.74%). Measured purity of the bulk

chemical was consistent throughout the sampled metal

drum, and the overall purity of lot 106

- 015 was deter

mined to be approximately 99%.

For lot 1581313004, Kar l Fischer titration indicated a

water content of 119 ppm. Elemental analyses for car

bon, hydrogen, nitrogen, and sulfur were in agreement

with the theoretical values for 1

- bromopropane; residual

HBr was determined to be 61.6 ppm. G C detected one

major peak and no impurities with areas greater than 0.1% of the total peak area. Th re e impurities identified

with prepared standards were 1- propanol (0.03%),

2- bromopropane (0.02%), and di

- n

- propyl ether

(0.02%). Measured purity of the bu lk chemical was

consistent throughout the sampled metal drums, and the

overall purity of lot 1581313004 was determined to be

99. 9 % or greater

.

To ensure stability, the bulk chemical was stored at

controlled room temperature in the original shippin g

contai ners (55

- gallon metal drums). Periodic reanalyses

of the bulk chemical were performed during the 2

- week,

3- month, and 2

- year studies by the study laboratory

using GC , and no degradation of the bulk chemical was

detected.



# V APOR 

# G ENERATION

AND 

# E XPOSURE 

# S YST EM

A preheater was necessary for the 2

- week and 3

- month

studies. 1- Bromopropane was pumped through a pre

heater and into a heated glass column filled with glass

beads that increased the surface area for vaporization.

Heated nitrogen entered the column fr om below and

assisted in vaporizing the chemical while conveying it

into a short distribution manifold. Concentration in the

manifold was determined by the chemical pump rate

and nitrogen flow rate. The pressure in the distribution

manifold was kept fixe d to ensure constant flow through

the manifold and into all chambers as the flow of vapor

to each chamber was adjusted.

Metering valves at the manifold controlled flow to each

chamber through individual Teflon ® 

### delivery lines that

carried the vapor from the manifold to three

- way expo

sure valves at the chamber inlets. The exposure valves

diverted vapor delivery to exposure chamber exhaust

until the generation system was stable and exposures

were ready to procee d. To initiate exposure, the cham

ber exposure valves were rotated to allow the 1

- bromo

propane vapor to flow to each exposure chamber inlet

duct where it was further diluted with filtered, condi

tioned air to achieve the desired exposure concentration.

### 26 1- Bromopropane , NTP TR 564

T he study laboratory designed the inhalation exposure

chamber (Harford Systems Division of Lab Products,

Inc., Aberdeen, MD) so that uniform vapor concentra

tions could be maintained throughout the chamber with

the catch pans in place. The total active mix ing volume

of each chamber was 1.7 m 3 

### . A small particle detector

was used with and without animals in the exposure

chambers to ensure that 1- bromopropane vapor, and not

aerosol, was produced. No particle counts above the

minimum resolvable level (approxi mately 200 particles

per cm 3 

### ) were detected

.



# V APOR 

# C ONCENTRATION 

# M ONITORING

Chamber and room concentrations of 1

- bromopropane

were monitored by an on

- line gas chromatograph.

Samples were drawn from each exposure chamber

approximately every 20 (2

- week and 3- month studies)

or 30 (2

- year studies) minutes during each 6

- hour expo

sure period using Hastelloy ®



### - C stream

- select and gas

sampling valves in a separate, heated valve oven. The

sample lines composing each sample loop were made

from Teflon ® 

### tubing and we re connected to the expo

sure chamber relative humidity sampling lines at a loca

tion close to the gas chromatograph. A vacuum regula

tor maintained a constant vacuum in the sample loop to

compensate for variations in sample line pressure. An

in

- line flo w meter between the vacuum regulator and

the gas chromatograph allowed digital measurement of

sample flow. Summaries of the chamber vapor concen

trations are given in Tables I2 through I4.

The on

- line gas chromatograph was checked throughout

the day for instrument drift against an on

- line standard

vapor of 1

- bromopropane in nitrogen supplied by a

standard generator. The on

- line gas chromatograph was

recalibrated as required to meet acceptance criteria.

Calibration was performed by comparing chamber con

centration data to data from grab samples that were

collected with activated coconut charcoal gas sampling

tubes, extracted with methylene chloride containing

1- bromobutane as an internal standard, and analyzed

using an off

- line gas chromatograph. Known v olumes

of chamber atmosphere were sampled at a constant flow

rate ensured by a calibrated critical orifice. The off

- line

gas chromatograph was calibrated with gravimetrically

prepared standards of 1

- bromopropane containing

1- bromobutane as an internal sta ndard in methylene

chloride. 

# C HAMBER 

# A TMOSPHERE



# C HARACTERIZATION

Buildup and decay rates for chamber vapor concentra

tions were determined with and without animals present

in the chambers. At a chamber airflow rate of 15 air

changes per hour, the theoretical value for the time to

achieve 90% of the target concentration after the begin

ning of vapor generation (T 90 

### ) and the time for the

chamber concentration to decay to 10% of the target

concentration after vapor generation was terminated

(T 10 

### ) was approximately 12.5 minutes. A T 90 

### value of

12 minutes was selected for the 2

- week studies, and a

T 90 

### value of 10 minutes was selected for the 3

- month

and 2

- year studies.

Evaluations of chamber uniformity and persistence and

monitoring for 1

- bromopropane degradat ion impurities

were conducted periodically throughout the studies by

GC. Chamber uniformity was maintained and no degra

d ation was detected.



# 2-W EEK 

# S TUDIES

Male and female F344/N rats and B6C3F1 mice were

obtained from Taconic Farms, Inc. ( Germantown

, NY ).

On receipt, the rats and mice were approximately

4 weeks old. Animals were quarantined for 13 days and

were approximately 5 to 6 weeks old on the first day of

the studies. Before the studies began, 10 female rats

and five m ale and five female mice we re rando mly

selected for parasite evaluation and gross observation

for evidence of disease. At terminal sacrifice, serologic

analyses were performed on five male and five female

chamber control rats and mice using the protocols of the

NTP Sentinel Animal Program (Appendix K ).

Groups of five male and five female rats and mice were

exposed to 1- bromopropane vapor at concentrations of

0, 125

, 250

, 500

, 1,000 , or 2,000 ppm, 6 hours plus T 90 

### ( 12 minutes) per day, 5 days per week for 16 (rats) or 17

(mice) days

. Feed was available ad libitum except dur

ing exposure periods; water w as available ad libitum

. Rats and mice were housed individually . Clinical find

ings were recorded twice daily on exposure days for rats

and mice. The animals were weighed initially, on

days 6 and 13 , and at the end of the studies. Details of

the study design and animal maintenance are summa

rized in Table 1.

### 1- Bromopropane, NTP TR 564 27

Necropsies were performed on all animals . The heart,

right kidney, li ver, lung, right testis, and thymus were

weighed. Tissues for microscopic examination were

fixed and preserved in 10% neutral buffered formalin

(except eyes were first fixed in Davidson’s solution)

, processed and trimmed, embedded in paraffin, sectioned

t o a thickness of 4 to 6 μ m, and stained with hematox

ylin and eosin. H istopathologic examinations were per

formed on all chamber control and 2,000 ppm animals

. The lung and nose of rats and mice; the mediastinal

lymph node, sciatic nerve, and spinal cord of rats; and

the liver of mice were examined to a no

- effect level

. Table 1 lists the tissues and organs examined.



# 3-M ONTH 

# S TUDIES

The 3

- month studies were conducted to evaluate the

cumulative toxic effects of repeated exposure to 1- bro

mopropane and to d etermine the appropriate exposure

concentrations to be used in the 2

- year studies.

Male and female F344/N rats and B6C3F1 mice were

obtained from Taconic Farms, Inc. ( Germantown

, NY )

. On receipt, the rats and mice were approximately 3 to

4 weeks old. Animals were quarantined for 12 (female

rats and male and female mice) or 13 (male rats) days

and were approximately 5 to 6 weeks old on the first

day of the studies. Before the studies began, five male

and five female rats and mice were randomly s elected

for parasite evaluation and gross observation for evi

dence of disease. Serological analyses were performed

on five male and five female sentinel rats and mice

during week 1 and five male and five female chamber

control rats and mice at the end of the studies using the

protocols of the NTP Sentinel Animal Program

(Appendix K ).

Groups of 10 male and 10 female rats and mice were

exposed to 1- bromopropane vapor at concentrations of

0, 62.5

, 125

, 250

, 500 , or 1,000 ppm (rat s only ), 6 hours

plus T 90 

### ( 10 minutes) per day, 5 days per week for

14 weeks. Additional clinical pathology groups of

10 male and 10 female rats were exposed to the same

concentrations for 23 days. One additional exposure

day was scheduled during the last exposure week to

give the rats at least 2 consecutive days of exposure

before terminal sacrifice. Feed was available ad libitum

except during exposure periods; water w as available

ad libitum . Rats and mice were housed individually

. Clinical findings were recorded on day 9 (male rats) or

10 and then weekly. Core study animals were weighed

initially, on day 9 (male rats) or 10, weekly thereafter

, and at the end of the studies. Details of the study design

and animal maintenance are summarized in Table 1.

Animals were anesthetize d with carbon dioxide, and

blood was collected from the retroorbital sinus of clinical

pathology rats on days 3 and 23 and from core study rats

at the end of the study for hematology and clinical

chem istry analyses; blood was collected from the retro

orbital sinus of mice at the end of the study for hema

tology analyses. Blood samples for hematology analyses

were placed in tubes containing potassium EDTA.

Erythrocyte, leukocyte, and platelet counts; hemoglobin;

packed red cell volume; mean cell volum e; mean cell

hemoglobin; and mean cell hemoglobin concentration

were determined using an Abbott Cell

- Dyn 3700 hema

tology analyzer (Abbott Diagnostic Systems, Abbott

Park, IL). Manual hematocrit values were determined

using a microcentrifuge (Heraeus Haemofuge, Germany)

and a Damon/IEC capillary reader (International

Equipment Company, Needham Heights, MA) for

comparison to Cell

- Dyn values for packed cell volume.

Blood smears for rats and mice were stained with

Romanowsky

- type aqueous stain in a Wescor 71 00

Aerospray Slide Stainer (Wescor, Inc., Logan, UT).

Leukocyte differential counts for rats and mice were

based on classifying a minimum of 100 white cells.

Reticulocytes were stained with N ew M ethylene B lue

and enumerated as a reticulocyte:erythrocyte ratio using

the Miller disc method (Brecher and Schneiderman,

1950). Blood samples for clinical chem istry analyses

were placed in tubes containing separator gel and

allowed to clot. After clot retraction occurred, the sam

ples were centrifuged, and the serum was aliquoted for

assay of serum chemistry analytes using a Roche Hitachi

912 (Roche Diagnostic Corporation, Indianapolis, IN).

Table 1 lists the parameters measured

.

At the end of the 3

- month studies, samples were col

lected for sperm motility and vaginal cytology evalua

tions on rats and mice exposed to 0, 125 (mice), 250,

500

, or 1,000 (rats) ppm . The parameters evaluated are

listed in Table 1. For 12 consecutive days prior to

scheduled terminal sacrifice, the vaginal vaults of the

females were moistened with saline, if necessary, and

samples of vaginal fluid and cells were stained. Rela

tive numbers of leukocytes, nucleated epithelial cells,

and large squamous epithelial cells were determined and

used to ascertain estrous cycle stage (i.e., die strus, pro

estrus, estrus, and metestrus). Male animals were evalu

ated for sperm count and motility. The left testis and

left epididymis were isolated and weighed. The tail of

the epididymis (cauda epididymis) was then removed

from the epididymal body (corpus epididymis) and

### 28 1- Bromopropane , NTP TR 564

weighed. Test yolk (rats) or modified Tyrode’s buffer

(mice) was applied to slides and a small incisio n was

made at the distal border of the cauda epididymis. The

sperm effluxing from the incision were dispersed in the

buffer on the slides, and the numbers of motile and

nonmotile spermatozoa were counted for five fields per

slide by two observers. Following completion of sperm

motility estimates, each left cauda epididymis was

placed in buffered saline solution. Caudae were finely

minced, and the tissue was incubated in the saline solu

tion and then heat fixed at 65° C. Sperm density was

then determined microscopically with the aid of a hema

cytometer. To quantify spermatogenesis, the testicular

spermatid head count was determined by removing the

tunica albuginea and homogenizing the left testis in

phosphate

- buffered saline containing 10% dimethyl

sulfo xide. Homogenization

- resistant spermatid nuclei

were counted with a hemacytometer

.

Necropsies were performed on all core study animals.

The heart, right kidney, liver, lung, spleen, right testis,

and thymus were weighed. Tissues for microscopic

examination were fixed and preserved in 10% neutral

buffered formalin (eyes were fixed in Davidson’s solu

tion for up to 3 days and then transferred to 10% neutral

buffered formalin) , processed and trimmed, embedded

in paraffin, sectioned to a thickness of 4 to 6 μm, and

stained with hematoxylin and eosin. Complete histo

pathologic examinations were performed on all chamber

control, 250 and 500 ppm mice, and 1,000 ppm rats

. The larynx, liver, lung, and nose of rats and mice, the

prostate gland of male rats, the trache a of mice, and the

adrenal gland and kidney of female mice were examined

in all groups; the remaining tissues were examined to a

no

- effect level in the remaining exposure groups

. Table 1 lists the tissues and organs routinely examined.

After a review of the laboratory reports and selected his

topathology slides by a quality assessment pathologist,

the findings and reviewed slides were submitted to a

NTP Pathology Working Group (PWG) coordinator for

a second independent review. Any inconsistencies in

the diagnoses made by the laboratory and quality assess

ment pathologists were resolved by the NTP pathology

peer review process. Final diagnoses for reviewed

lesions represent a consensus between the laboratory

pathologist, NTP pathologist, reviewing pathologist(s),

if any, and the PWG coordinator. Details of these

review procedures have been described, in part, by

Maronpot and Boorman (1982) and Boorman et al

. (1985). 

# 2-Y EAR 

# S TUDIES



## Study Design

Groups of 50 male and 50 female rats an d mice were

exposed to 1- bromopropane vapor at concentrations of

0, 62.5 (mice only)

, 125

, 250 , or 500 (rats only) ppm,

6 hours plus T 90 

### ( 10 minutes) per day, 5 days per week

for up to 105 weeks

.



## Source and Specification of Animals

Male and female F344/N rats and B6C3F1 mice were

obtained from Taconic Farms, Inc. ( Germantown

, NY ),

for use in the 2

- year studies. Rats were quarantined for

13 days and mice were quarantined for 11 days before

the beginning of the studies. Five male and five female

rats and 10 male mice were randomly selected for para

site evaluation and gross observation of disease. Rats

and mice were approximately 5 to 6 weeks old a t the

beginning of the studies. The health of the animals was

monitored during the studies according to the protocols

of the NTP Sentinel Animal Program (Appendix K ).



## Animal Maintenance

Rats and mice were housed individually. Feed was avail

able ad libitum except during exposure periods; water

w as available ad libitum . Cages

, racks , and chambers

were changed weekly. Cages were rotated weekly in

chambers . Further details of animal maintenance are

given in Table 1. Information on feed composition and

contaminants is provided in Appendix J.



## Clinical Examinations and Pathology

All animals were observed twice d aily. Clinical find

ings were recorded for all animals every 4 weeks

through week 93, every 2 weeks thereafter, and at the

end of the studies. Rats and mice were weighed ini

tially, weekly for the first 13 weeks, then every 4 weeks

through week 93, every 2 weeks thereafter, and at the

end of the studies

.

Complete necropsies and microscopic examinations

were performed on all rats and mice. At necropsy, all

organs and tissues were examined for grossly visible

lesions, and all major tissues were fixed and preserved

in 10% neutral buffered formalin (eyes were fixed in

Davidson’s solution for up to 72 hours and then trans

ferred to 10% neutral buffered formalin) , processed and

trimmed, embedded in paraffin, sectioned to a thickness

of 4 to 6 μm, and stained with hematoxylin and eosin

for microscopic examination. For all paired organs

(e.g., adrenal gland, kidney, ovary), samples from each

### 1- Bromopropane, NTP TR 564 29

organ were examined. Tissues examined microscopi

cally are listed in Table 1.

Microscopic evaluations were completed by the study

laboratory pathologist, and the pathology data were

entered into the Toxicology Data Management System.

The reports, slides, paraffin blocks, residual wet tissues

, and pathology data were sent to the NTP Archives for

inventory, slide/block matc h, wet tissue audit , and

storage . The slides, individual animal data records, and

pathology tables were evaluated by an independent

quality assessment laboratory. The individual animal

records and tables were compared for accuracy, the

slide and tissue c ounts were verified, and the histotech

nique was evaluated. For the 2

- year studies, a quality

assessment pathologist evaluated slides from all tumors

and all potential target organs, which included the

larynx, nose, and trachea of rats and mice and the lu ng

of mice

.

The quality assessment report and the reviewed slides

were submitted to the NTP PWG coordinator, who

reviewed the selected tissues and addressed any incon

sistencies in the diagnoses made by the laboratory and

quality assessment pathologists. Representative histo

pathology slides containing examples of lesions related

to chemical administration, examples of disagreements

in diagnoses between the laboratory and quality assess

ment pathologists, or lesions of general interest were

presented by t he coordinator to the PWG for review. The PWG consisted of the quality assessment pathol

ogist and other pathologists experienced in rodent toxi

cologic pathology. This group examined the tissues

without any knowledge of dose groups. When the PWG

conse nsus differed from the opinion of the laboratory

pathologist, the diagnosis was changed. Final diagnoses

for reviewed lesions represent a consensus between the

laboratory pathologist, reviewing pathologist(s), and the

PWG. Details of these review procedures have been

described, in part, by Maronpot and Boorman (1982)

and Boorman et al. (1985). For subsequent analyses of

the pathology data, the decision of whether to evaluate

the diagnosed lesions for each tissue type separately or

combined was generally based on the guidelines of

McConnell et al. (1986).

Three standard sections are taken through the nose in

NTP studies, and these are referred to as Levels I, II,

and III. Proceeding from anterior to posterior, Level I is

taken immediately posterior to the upper incisor teeth;

Level II is taken through the level of the incisive papill a

anterior to the first palatal ridge; and Level III is taken

through the middle of the second molar teeth (Figure 2).

The mucosa of the nasal passages in Levels I and II is

lined by respiratory and transitional epithelium, except

for the ventral meatus o f Levels I and II (squamous

epithelium) and the dorsal meatus of Level II (olfactory

epithelium). Level III is lined almost entirely by olfac

tory epithelium, except for the ventral meatus, which is

lined by respiratory epithelium.

### 30 1- Bromopropane , NTP TR 564

F IGURE 

### 2

Rodent Nasal Cavity Diagram Illustrating the Levels of Sections

Level I: Immediately posterior to incisor teeth

Level II: At incisive papilla anterior to first palatal ridge

Level III: Section taken through the second molar

MT=maxilloturb inate; NT=nasoturbinate; ET=ethmoid turbinates

### 1- Bromopropane, NTP TR 564 31

T ABLE 

### 1

Experimental Design and Materials and Methods in the Inhalation Studies of 1- Bromopropane

2- Week Studies 3- Month Studies 2- Year Studies

Study Laboratory

Battelle Toxicology Northwest

(Richland, WA) Battelle Toxicology Northwest

(Richland, WA) Battelle Toxicology Northwest

(Richland, WA)



### Strain and Species

F344/N rats F344/N rats F344/N rats

B6C3F1 mice B6C3F1 mice B6C3F1 mice



### Animal Source

Taconic Farms, Inc. (Germantown, NY) Taconic Farms, Inc. (Germantown, NY) Taconic Farms, Inc. (Germantown, NY)



### Time Held Before Studies

1 3 days Rats: 1 2 ( fe males) or 1 3 (males) days Rats: 13 days

Mice: 12 days Mice: 11 days



### Average Age When Studies Began

5 to 6 weeks 5 to 6 weeks 5 to 6 weeks



### Date of First Exposure

July 15, 2002 Rats: November 1 1 ( fe males) or 1 2 (males),

2002

Mice: November 11, 2002 Rats: July 14, 2003

Mice: July 21, 2003



### Duration of Exposure

6 hours plus T 90 (12 minutes) per day, 5 days

per week, for 16 (rats) or 17 (mice) days 6 hours plus T 90 (10 minutes) per day, 5 days

per week, for 14 weeks 6 hours plus T 90 (10 minutes) per day, 5 days

per week, for 1 05 weeks



### Date of Last Exposure

Rats: July 30, 2002

Mice: July 31, 2002 Rats: February 1 0 ( fe males) or 1 1 ( m ales),

2003

Mice: February 12 (males) or 13 (females),

2003 Rats: July 1 3 , 2005

Mice: July 21, 2005



### Necropsy Dates

Rats: July 31, 2002

Mice: August 1, 2002 Rats: February 1 1 ( fe males) or 1 2 (males),

2003

Mice: February 13 (males) or 14 (females),

2003 Rats: July 11

- 14, 2005

Mice: July 18

- 22, 2005



### Average Age at Necropsy

8 to 9 weeks 18 to 19 weeks Rats: 110 weeks

Mice: 109 to 111 weeks



### Size of Study Groups

5 males and 5 females 10 males and 10 females 50 males and 50 females



### Method of Distribution

Animals were distributed randomly into

groups of approximately equal initial mean

body weights. Same as 2

- week studies Same as 2

- week studies



### Animals per Cage

1 1 1



### Method of Animal Identification

Tail tattoo Tail tattoo Tail tattoo

### 32 1- Bromopropane , NTP TR 564

T ABLE 

### 1

Experimental Design and Materials and Methods in the Inhalation Studies of 1

- Bromopropane

2- Week Studies 3- Month Studies 2- Year Studies

Diet

NTP

- 2000 irradiated pelleted diet (Zeigler

Brothers, Inc., Gardners, PA) ; available

ad libitum , except during exposure periods ;

changed weekly Same as 2

- week studies Same as 2

- week studies



### Water

Tap water ( Richland, WA, municipal supply)

via automatic watering system ( Edstrom

Industries, Waterford

, WI ); available

ad libitum Same as 2

- week studies Same as 2

- week studies



### Cages

Stainless steel, wire bottom ( Lab Products,

Inc.

, Seaford, DE); changed weekly , rotated

daily Same as 2

- week studies , except rotated

weekly Same as 3

- month studies



### Cageboard

Untreated paper cage pan liner ( Sheperd

Specialty Papers , Kalamazoo, MI ); changed

daily Same as 2

- week studies Same as 2

- week studies



### Chamber Air Supply Filters

Single HEPA ( open stock )

, charcoal (RSE,

Inc., New Baltimore, MI)

, Purafil

(Environmental Systems, Lynnwood, WA),

all new at study start Same as 2

- week studies , except not changed

after 2

- week studies Same as 2

- week studies , except single HEPA

changed annually



### Chambers

Stainless steel, excreta pan at each of six

levels (Lab Products, Inc.

, Seaford

, DE );

chambers changed weekly; excreta pans

changed daily Same as 2

- week studies Same as 2

- week studies



### Chamber Environment

Temperature: 72° ± 3° F Temperature: 72° ± 3° F Temperature: 72° ± 3° F

Relative humidity: 50% ± 15% Relative humidity: 50% ± 15% Relative humidity: 50% ± 15%

Room fluorescent light: 12 hours/day Room fluorescent light: 12 hours/day Room fluoresc ent light: 12 hours/day

Chamber air changes: 15 ± 2 /hour Chamber air changes: 15 ± 2 / hour Chamber air changes: 15 ± 2 /hour



### Exposure Concentrations

0, 125, 250, 500, 1,000, or 2,000 ppm Rats: 0, 62.5, 125, 250, 500, or 1,000 ppm

Mice: 0, 62.5, 125, 250, or 500 ppm Rats: 0, 125, 250, or 500 ppm

Mice: 0, 62.5, 125, or 250 ppm



### Type and Frequency of Observation

Observed twice daily; animals were weighed

initially, on days 6 and 13, and at the end of

the studies; clinical findings were recorded

twice daily on exposure days. Observed twice daily; core study animals

were weighed initially, on days 9 (male rats)

or 10, weekly thereafter , and at the end of the

studies

. C linical findings were recorded on

day 9 (male rats) or 10 and then weekly

. Observed twice daily; animals were weighed

initially, weekly for 13 weeks, every 4 weeks

through week 93, every 2 weeks thereafter,

and at the end of the studies; clinical findings

were recorded every 4 weeks through

week 93, every 2 weeks thereaf ter , and at the

end of the studies.

### 1- Bromopropane, NTP TR 564 33

T ABLE 

### 1

Experimental Design and Materials and Methods in the Inhalation Studies of 1

- Bromopropane

2- Week Studies 3- Month Studies 2- Year Studies

Method of Sacrifice

Carbon dioxide asphyxiation Same as 2

- week studies Same as 2

- week studies



### Necropsy

Necropsies were performed on all animals.

Organs weighed were heart, right kidney,

liver, lung, right testis, and thymus. Necropsies were performed on core study

animals. Organs weighed were heart, right

kidney, liver, lung, spleen

, right testis, and

thymus

. Necropsies were performed on all animals.



### Clinical Pathology

None Blood was collected from the retroorbital

sinus of clinical pathology rats on days 3 and

23 and from core study rat s at the end of the

studies for hematology and clinical chemistry.

Blood was collected from the retroorbital

sinus of mice at the end of the study for

hematology. None

Hematology: hematocrit; packed red cell

volume; hemoglobin; erythrocyte, nucleate d

erythrocyte, reticulocyte, and platelet counts;

mean cell volume; mean cell hemoglobin;

mean cell hemoglobin concentration; and

leukocyte count and differentials

Clinical chemistry: urea nitrogen, creatinine,

total protein, albumin, globulin, alanine

aminotransferase, alkaline phosphatase,

creatine kinase, sorbitol dehydrogenase, and

bile acids



### Histopathology

H istopathology was performed on 0 and

2,000 ppm rats and mice. In addition to gross

lesions and tissue masses, the following

tissues were examined: lung, mediastinal

lymph node, n ose , sciatic nerve, and spinal

cord of rats and liver, lung, and nose of mice.

These tissues were examined to a no

- effect

level in the remaining exposure groups. Complete histopathology was performed on 0

and 1,000 ppm core study rats and 0, 250, and

500 ppm mice. In addition to gross lesions

and tissue masses, the following tissues were

examined to a no

- effect level : adrenal gland,

bone with marrow, brain, clitoral gland,

esophagus, eyes, gallbladder (mice)

, Harderian gland, heart and aorta, large

intestine (cecum, colon, rectum), small

intestine (duodenum, jejunum, ileum), kidney,

larynx, liver, lung, lymph nodes (mandibular,

mesenteric, bronchial, mediastinal),

mammary gland, nose, ovary, pancreas,

parathy roid gland, pituitary gland, preputial

gland, prostate gland, salivary gland, skin,

spleen, stomach (forestomach and glandular),

testis with epididymis and seminal vesicle,

thymus, thyroid gland, trachea, urinary

bladder, and uterus. In addition, the live r,

larynx, lung, and nose of rats and mice; the

trachea of mice; the prostate gland of male

rats; and the adrenal gland and kidney of

female mice were examined in the remaining

groups. Complete histopathology was performed on

all rats and mice. In additio n to gross lesions

and tissue masses, the following tissues were

examined: adrenal gland, bone with marrow,

brain, clitoral gland, esophagus, eye,

gallbladder (mice only), Harderian gland,

heart with aorta, large intestine (cecum, colon,

rectum), small in testine (duodenum, jejunum,

ileum), kidney, larynx, liver, lung, lymph

nodes (mandibular, mesenteric, bronchial,

mediastinal), mammary gland, nose, ovary,

pancreas, pancreatic islets, parathyroid gland,

pituitary gland, preputial gland, prostate

gland, salivary gland, skin, spleen, stomach

(forestomach and glandular), testis with

epididymis and seminal vesicle, thymus,

thyroid gland, trachea, urinary bladder, and

uterus.

### 34 1- Bromopropane , NTP TR 564

T ABLE 

### 1

Experimental Design and Materials and Methods in the Inhalation Studies of 1- Bromopropane

2- Week Studies 3- Month Studies 2- Year Studies

Sperm Motility

and Vaginal Cytology

None At the end of the studies, sperm samples were

collected from male animals in the 0

, 125

(mice)

, 250

, 500, and 1,000 (rats) ppm groups

for sperm motility evaluations. The following

parameters were evaluated: spermatid heads

per testis and per gram testis, spermatid

counts, and epididymal spermatozoal motility

and concentration. The left cauda, left

epididymis, and left testis were weig hed.

Vaginal samples were collected for up to

12 consecutive days prior to the end of the

studies from females exposed to 0, 125

(mice), 250, 500, or 1,000 (rats) ppm for

vaginal cytology evaluations. The percentage

of time spent in the various estrous c ycle

stages and estrous cycle length were

evaluated. None



# S TATISTICAL 

# M ETHODS



## Survival Analyses

The probability of survival was estimated by the prod

uct

- limit procedure of Kaplan and Meier (1958) and is

presented in the form of graphs. Animals found dead of

other than natural causes were censored; animals dying

from natural causes were not censored. Statistical anal

yses for possible dose

- related effects on survival used

Cox’s (1972) method for testing two groups for equality

and Taron e’s (1975) life table test to identify dose

related trends. All reported P values for the survival

analyses are two sided.



## Calculation of Incidence

The incidences of neoplasms or nonneoplastic lesions

are presented in Tables A1, A 4 , B1, B 4 , C1, C 3 , D1,

and D 4 as the numbers of animals bearing such lesions

at a specific anatomic site and the numbers of animals

with that site examined microscopically. For calcula

tion of statistical significance, the incidences of most

neoplasms (Tables A2, B2, C2, and D2 ) and all non

neoplastic lesions are given as the numbers of animals

affected at each site examined microscopically. How

ever, when macroscopic examination was required to

detect neoplasms in certain tissues (e.g., mesentery,

pleura, peripheral nerve, ske letal muscle, tongue, tooth,

and Zymbal’s gland ) before microscopic evaluation, the

denominators consist of the number of animals that had

a gross abnormality

. W hen neoplasms had multipl e potential sites of occurrence (e.g., leukemia or lym

phoma), the denominators consist of the number of

animals on which a necropsy was performed.

Tables A2, B2, C2, and D2 also give the survival

adjusted neoplasm rate for each group and each site

specific neoplasm. This survival

- adjusted rate (based

on the Poly

- 3 met hod described below) accounts for

differential mortality by assigning a reduced risk of

neoplasm, proportional to the third power of the fraction

of time on study, only to site

- specific, lesion

- free ani

mals that do not reach terminal sacrifice.



## Analysis of Neoplasm

and Nonneoplastic Lesion Incidences

The Poly

- k test (Bailer and Portier, 1988; Portier and

Bailer, 1989; Piegorsch and Bailer, 1997) was used to

assess neoplasm and nonneoplastic lesion prevalence.

This test is a survival

- adjusted quantal

- resp onse proce

dure that modifies the Cochran

- Armitage linear trend

test to take survival differences into account. More spe

cifically, this method modifies the denominator in the

quantal estimate of lesion incidence to approximate

more closely the total numb er of animal years at risk.

For analysis of a given site, each animal is assigned a

risk weight. This value is one if the animal had a lesion

at that site or if it survived until terminal sacrifice; if the

animal died prior to terminal sacrifice and did not have

### 1- Bromopropane, NTP TR 564 35

a lesion at that site, its risk weight is the fraction of the

entire study time that it survived, raised to the kth

power.

This method yields a lesion prevalence rate that depends

only upon the choice of a shape parameter for a Weibull

hazard fun ction describing cumulative lesion incidence

over time (Bailer and Portier, 1988). Unless otherwise

specified, a value of k=3 was used in the analysis of

site

- specific lesions. This value was recommended by

Bailer and Portier (1988) following an evaluati on of

neoplasm onset time distributions for a variety of site

specific neoplasms in control F344 rats and B6C3F1

mice (Portier et al. , 1986). Bailer and Portier (1988)

showed that the Poly

- 3 test gave valid results if the true

value of k was anywhere in t he range from 1 to 5. A

further advantage of the Poly

- 3 method is that it does

not require lesion lethality assumptions. Variation

introduced by the use of risk weights, which reflect dif

ferential mortality, was accommodated by adjusting the

variance of the Poly

- 3 statistic as recommended by

Bieler and Williams (1993).

Tests of significance included pairwise comparisons of

each exposed group with controls and a test for an over

all exposure

- related trend. Continuity

- corrected Poly

- 3

tests were used in the analysis of lesion incidence, and

reported P values are one sided. The significance of

lower incidences or decreasing trends in lesions is

represented as 1

– P with the letter N added (e.g., P=0.99

is presented as P=0.01N).



## Analysis of Continuous Varia bles

Two approaches were employed to assess the signifi

cance of pairwise comparisons between exposed and

control groups in the analysis of continuous variables.

Organ and body weight data, which historically have

approximately normal distributions, were analyzed with

the parametric multiple comparison procedu res of

Dunnett (1955) and Williams (1971, 1972). Hematol

ogy, clinical chemistry, spermatid, and epididymal

spermatozoal data, which have typically skewed distri

butions, were analyzed using the nonparametric mul

tiple comparison methods of Shirley (1977) (as mod

ified by Williams, 1986) and Dunn (1964).

Jonckheere’s test (Jonckheere, 1954) was used to assess

the significance of the dose

- related trends and to deter

mine whether a trend

- sensitive test (Williams’ or

Shirley’s test) was more appropriate for pairwise com

parisons than a test that does not assume a monotonic

dose

- related trend (Dunnett’s or Dunn’s test). Prior to

statistical analysis, extreme values identified by the out

lier test of Dixon and Massey (1957) were examined by NTP personnel, and implausible values were eliminated

from the analysis. Because vaginal cytology data are

proportions (the proportion of the observation period

that an animal was in a given estrous stage), an arcsine

transformation was used to bring the data into closer

co nformance with a normality assumption

. Treatment

effects were investigated by applying a multivariate

analysis of variance (Morrison, 1976) to the transformed

data to test for simultaneous equality of measurements

across exposure concentrations

. Proporti ons of regular

cycling females in each exposed group were compared

to the control group using the Fisher exact test (Gart

et al. , 1979). Tests for extended periods of estrus and

diestrus were constructed based on a Markov chain

model proposed by Girard and Sager (1987). For each

exposure group, a transition probability matrix was

estimated for transitions among the proestrus, estrus,

metestrus, and diestrus stages, with provision for

extended stays within estrus and diestrus. Equality of

transition matri ces among exposure groups and between

the control group and each exposed group was tested

using chi

- square statistics.



## Historical Control Data

The concurrent control group represents the most valid

comparison to the treated groups and is the only control

group analyzed statistically in NTP bioassays. How

ever, historical control data are often helpful in inter

preting potential treatment

- related effects, particularly

for uncommon or rare neoplasm types. For meaningful

comparisons, the conditions for stud ies in the historical

database must be generally similar. One significant

factor affecting the background incidence of neoplasms

at a variety of sites is diet. In 1995, the NTP incorpo

rated a new diet (NTP

- 2000) that contains less protein

and more fiber and fat than the NIH

- 07 diet previously

used in toxicity and carcinogenicity studies (Rao, 1996,

1997). The NTP historical database contains all studies

that use the NTP

- 2000 diet with histopathology findings

completed within the most recent 5

- year perio d. A

second potential source of variability is route of admin

istration. In general, the historical database for a given

study will include studies using the same route of

administration, and the overall incidences of neoplasms

for all routes of administ ration are included for com

parison, including the present study.



# Q UALITY 

# A SSURANCE 

# M ETHODS

The 3- month and 2

- year studies were conducted in

compliance with Food and Drug Administration Good

Laboratory Practice Regulations (21 CFR, Part 58). In

### 36 1- Bromopropane , NTP TR 564

addition, as records from the 2

- year studies were sub

mitted to the NTP Archives, these studies were audited

retrospectively by an independent quality assurance

contractor. Separate audits covered completeness and

accuracy of the pathology data, pathology specimen s,

final pathology tables, and a draft of this NTP Technical

Report. Audit procedures and findings are presented in

the reports and are on file at NIEHS. The audit findings

were reviewed and assessed by NTP staff, and all com

ments were resolved or other wise addressed during the

preparation of this Technical Report.



# G ENETIC 

# T OXICOLOGY

The genetic toxicity of 1- bromopropane was assessed

by testing the ability of the chemical to induce muta

tions in various strains of Salmonella typhimurium and

Escherichia coli and increases in the frequency of

micronucleated erythrocytes in mouse peripheral blood.

Micronuclei (literally “small nuclei” or Howell

- Jolly

bodies) are biomarkers of induced structural or numer

ical chromosomal alterations and are formed when

acen tric fragments or whole chromosomes fail to incor

porate into either of two daughter nuclei during cell

division (Schmid, 1975; Heddle et al. , 1983). The pro

tocols for these studies and the results are given in

Appendix E.

The genetic toxicity studies h ave evolved from an ear

lier effort by the NTP to develop a comprehensive data

base permitting a critical anticipation of a chemical’s

carcinogenicity in experimental animals based on

numerous considerations, including the molecular

structure of the chemic al and its observed effects in

short

- term in vitro and in vivo genetic toxicity tests

(structure

- activity relationships). The short

- term tests

were originally developed to clarify proposed mechan

isms of chemical

- induced DNA damage b ased on the

relationsh ip between electrophilicity and mutagenicity

(Miller and Miller, 1977) and the somatic mutation theory of cancer (Straus, 1981; Crawford, 1985). How

ever, it should be noted that not all cancers arise through

genotoxic mechanisms.

DNA reactivity combine d with Salmonella mutagenicity

is highly correlated with induction of carcinogenicity in

multiple species/sexes of rodents and at multiple tissue

sites (Ashby and Tennant, 1991). A positive response

in the Salmonella test was shown to be the most predic

t ive in vitro indicator for rodent carcinogenicity (89% of

the Salmonella mutagens are rodent carcinogens)

(Tennant et al. , 1987; Zeiger et al. , 1990). Additionally,

no battery of tests that included the Salmonella test

improved the predictivity of the Sal monella test alone.

However, these other tests can provide useful informa

tion on the types of DNA and chromosomal damage

induced by the chemical under investigation.

The predictivity for carcinogenicity of a positive

response in acute in vivo bone marrow chromosome

aberration or micronucleus tests appears to be less than

that in the Salmonella test (Shelby et al. , 1993; Shelby

and Witt, 1995). However, clearly positive results in

long

- term peripheral blood micronucleus tests have high

predict ivity for rodent carcinogenicity; a weak response

in one sex only or negative results in both sexes in this

assay do not correlate well with either negative or posi

tive results in rodent carcinogenicity studies (Witt et al.

, 2000) . Because of the theoretical and observed associ

ations between induced genetic damage and adverse

effects in somatic and germ cells, the determination of

in vivo genetic effects is important to the overall under

standing of the risks associated with exposure to a par

ticular che mical. Most organic chemicals that are iden

tified by the International Agency for Research on

Cancer as human carcinogens, other than hormones, are

genotoxic. The vast majority of these are detected by

both the Salmonella assay and rodent bone marrow

cy togenetics tests (Shelby, 1988; Shelby and Zeiger,

1990).

### 37



# RESULTS

R ATS



# 2-W EEK 

# S TUDY

Except for one 500 ppm male that died on day 14 , all

rats survived to the end of the stud y (Table 2). Final

mean body weights and body weight gains of 2 ,000 ppm

rats were significantly less than those of the chamber

controls, as w as the body weight gain of 1,000 ppm

males. The neurological sign of hind limb splaying was

observed in some 2,000 ppm rat s after 1 week of expo

sure to 1

- bromopropane

. This was a transient change

and the animals showed marked improvement after a

day. The absolute weights of right kidney in 1,000 ppm

males and all exposed groups of females were

significantly greater than those of the chamber controls

(Table G1). The relative right kidney weights were

significantly increased in all exposed groups of both

sexes. Absolute liver weights were significantly

increased in 1,000 ppm males and 500 ppm or greater

females. Rel ative liver weights were significantly

increased in 500 ppm or greater males and all exposed

groups of females.

T ABLE 

### 2

Survival and Body Weights of Rats in the 2

- Week Inhalation Study of 1

- Bromopropane

Concentration

(ppm) Survival a 

### Mean Body Weight b 

### (g) Final Weight

Relative to Controls

(%) Initial Final Change

Male

0 5/5 99 ± 4 16 9 ± 4 69 ± 3

125 5/5 99 ± 4 1 67 ± 4 68 ± 1 99

250 5/5 98 ± 4 16 9 ± 5 70 ± 3 100

5 00 4/5 c 98 ± 4 1 66 ± 6 69 ± 1 9 8

1,0 00 5 /5 102 ± 4 1 64 ± 5 63 ± 2* 97

2,0 00 5/5 98 ± 4 1 2 3 ± 4** 25 ± 2 ** 73



### Female

0 5/5 87 ± 3 1 2 0 ± 2 33 ± 1

125 5/5 88 ± 2 1 24 ± 3 37 ± 2 104

250 5/5 86 ± 3 12 0 ± 4 34 ± 3 100

500 5/5 89 ± 3 1 24 ± 5 35 ± 3 104

1,000 5/5 88 ± 2 1 19 ± 2 31 ± 1 99

2,0 00 5/5 89 ± 2 1 07 ± 2* 18 ± 1** 89

* Significantly different (P ≤ 0.05) from the chamber control group by Williams’ or Dunnett’s t est

** P ≤ 0.01

a Number of animals surviving at 2 weeks /number initially in group

b Weights and weight changes are given as mean ± standard error. Subsequent calculations are based on animals surviving to the end of the

study.

c Day of death: 14

### 38 1- Bromopropane, NTP TR 564

M icroscopic lesions related to 1

- bromopropane expo

sure occurred in the nose. I ncidences of minima l to

mild suppurative inflammation occurred in males

exposed to 500 ppm or greater, incidences of minimal

respiratory epithelium necrosis occurred in 1,000 and

2,000 ppm males, and incidences of minimal respiratory

epithelium regeneration occurred in females exposed to

1,000 and 2,000 ppm (Table 3). Sciatic nerve and spinal cord were examined micro

scopically, but no lesions were identified.

Exposure Concentration Selection Rationale : Based on

decreased body weights and s igns of neurotoxicity at

2,000 ppm in the 2

- week study, 1

- bromopropane expo

sure concentrations selected for the 3

- month inhalation

study in rats were 62.5, 125, 250, 500, and 1,000 ppm.

T ABLE 

### 3

Incidences of Nonneoplastic Lesions in the Nose o f Rats in the 2

- Week Inhalation Study of 1

- Bromopropane

Chamber

Control 12 5 ppm 2 50 ppm 500 ppm 1,000 ppm 2 ,000 ppm

Male

Number Examined Microscopically 5 0 5 5 5 5

Respiratory Epithelium,

Necrosis a 0 0 0 2 ( 1 .0) b 1 ( 1.0 )

Suppurative Inflammation 0 0 1 (1.0) 2 (1.5) 2 (1.5)



### Female

Number Examined Microscopically 5 0 0 5 5 5

Respiratory Epithelium,

Regeneration 0 0 1 ( 1 .0) 1 ( 1

. 0 )

a Number of animals with lesion

b Average severity grade of lesions in affected animals: 1=minimal, 2=mild, 3=moderate, 4=marked

### 1- Bromopropane, NTP TR 564 39



# 3-M ONTH 

# S TUD Y

All rats survived to the end of the study (Table 4). The

final mean body weight and mean body weight gain of

1,000 ppm males were significantly less than those of

the chamber controls (Table 4 and Figure 3)

. Mean

b ody weights of exposed females were similar to those

of the chamber controls.

There were no changes in hematology endpoints that

were considered related to 1- bromopropane exposure

(Table F1). There were early, transient decreases in

albumin and total protein concentrat ions and alanine

aminotransferase activities in most exposed groups of

male and female rats. These transient decreases may

have been associated with effects of 1

- bromopropane on

hepatic protein metabolism. Additionally, sorbitol dehydrogenase (SDH) act ivit y was increase d at day 23

and at the end of the study in 1,000 ppm females and at

the end of the study in 500 and 1,000 ppm males.

Increased SDH activity would be consistent with mild

hepatotoxicity caused by 1

- bromopropane.

The absolute and relative liver weights of males

exposed to 250 ppm or greater and females exposed to

125 ppm or greater were significantly increased

(Table G2). The absolute spleen weights of females

exposed to 125 ppm or greater and the relative spleen

weight of 1,000 ppm femal es were greater than those of

the chamber controls. In addition, the absolute and rel

ative right kidney weights of 1,000 ppm females were

greater than those of the chamber controls.

T ABLE 

### 4

Survival and Body Weights o f Rats in the 3

- Month Inhalation Study of 1

- Bromopropane

Concentration

(ppm) Survival a 

### Mean Body Weight b 

### ( g ) Final Weight

Relative to Controls

(%) Initial Final Change

Male

0 10 /10 99 ± 2 345 ± 7 246 ± 6

62.5 10/10 102 ± 2 346 ± 5 244 ± 5 100

125 10/10 100 ± 2 348 ± 6 248 ± 5 101

250 10 /10 101 ± 3 350 ± 7 250 ± 6 101

500 10 /10 99 ± 2 344 ± 10 245 ± 10 100

1,000 10 /10 100 ± 2 305 ± 6** 205 ± 4** 88



### Female

0 10/10 88 ± 1 202 ± 3 114 ± 3

62.5 10/10 92 ± 3 203 ± 5 111 ± 3 101

125 10/10 88 ± 2 207 ± 4 120 ± 4 103

250 10/10 91 ± 2 209 ± 2 119 ± 3 104

500 10/10 86 ± 2 205 ± 3 119 ± 3 102

1,000 10 /10 88 ± 2 190 ± 4 102 ± 3 95

** Significantly different (P ≤ 0.0 1 ) from the chamber control group by Dunnett’s or Williams’ te st

a Number of animals surviving at 3 months /number initially in group

b Weights and weight changes are given as mean ± standard error.

### 40 1- Bromopropane , NTP TR 564

F IGURE 

### 3

Growth Cu rves for Rats Exposed to 1

- Bromopropane

by Inhalation for 3 Months

### 1- Bromopropane, NTP TR 564 41

There were significant exposure concentration

- related

decreases in sperm motility in male rats exposed to

250 ppm or greater (7%, 10%, and 2 8 % in the 250, 500,

and 1,000 ppm groups, respectively)

, and there were

significant decreases in the number of sperm per gram

cauda and the total sperm per cauda (25% and 37 %,

respectively) , as well as significant de creases in the

absolute weights of the cauda (14%) and left epidid

ymis (1 9 %) of 1,000 ppm male rats (Table H1). Female

rats in each of the exposure groups evaluated differe d

significantly from the chamber controls in the relative

amount of time spent in the various estrous cycle stages

, with each exposed group spending signifi cantly more

time in extended estrus and significantly less time in

extended diestrus (Table H2).

Treatment

- related lesions occurred in the liver. There

were significantly increased incidences of hepatocellu

lar cytoplasmic vacuolization in males exposed to

250 ppm or greater and in females exposed to 500 or

1,000 ppm (Table 5). The incidence of h epatocellular

degeneration was significantly increased in females

exposed to 1,000 ppm. Hepatocellular cytoplasmic

vacuolization consisted of swollen hepatocytes with

centrally located nuclei and one to three variably sized

vacuoles displacing the cytoplasm. H epatocellular vacuolar degeneration consisted of a distinct population

of enlarged, pale

- staining, degenerative, “ balloon

- like”

cells admixed with low numbers of necrotic hepa

tocytes. The lesion involved hepatocytes surrounding

the central vein, but in the most severe cases, the

vacuolated hepatocytes extended into the midzonal

region.

The incidence of minimal suppurative inflam mation of

the pr ostate in 1,000 ppm males was increased, but the

increase was not statistically significant (data not

shown). The increasing trend of this lesion, however,

was significant. Because this lesion is a common

background finding in F344/N rats, the biological

signifi cance of the increased incidence in the 1,000 ppm

males is unclear ( Boorman et al ., 1990 ).

Exposure Concentration Selection Rationale : Based on

decreased body weights and increased incidences of

liver lesions in the 3

- month study

, 1- bromopropane

exposure concentrations selected for the 2

- year inhala

tion study in rats were 125, 250, and 500 ppm.

Although 500 ppm caused vacuolization in liver, this

lesion was not considered life

- threatening and the use of

this concentration was not expected to co mpromise the

2- year study.

T ABLE 

### 5

Incidences of Nonneoplastic Lesions of the Liver in Rats in the 3

- Month Inhalation Study of 1

- Bromopropane

Chamber

Control 62.5 ppm 125 ppm 250 ppm 500 ppm 1,000 ppm

Male

Number Examined Microscopically 10 10 10 10 10 10

Vacuolization Cytoplasmic a 0 0 0 5* (1.0) b 10** (2.2) 10** (2.8)



### Female

Number Examined Microscopically 10 10 10 10 10 10

Vacuolization Cytoplasmic 0 0 0 0 10** (1.1) 10** (2.6)

Hepatocyte, Degeneration 0 0 0 0 0 7** (1.9)

* Significantly different (P ≤ 0.05) from the chamber control group by the Fisher exact test

** P ≤ 0.01

a Number of animals with lesion

b Average severity grade of lesions in affected animals: 1=minimal, 2=mild, 3=moderate, 4=marked

### 42 1- Bromopropane, NTP TR 564



# 2-Y EAR 

# S TUDY



## Survival

Estimates of 2

- year survival probabilities for male and

female rats are shown in Table 6 and in the Kaplan

Meier survival curves (Figure 4 ). Survival of 500 ppm

males was significantly less than that of the chamber

control group

. T he majority of the early deaths in the

500 ppm males w ere attribut ed to various types of

neoplasia, none of which w ere treatment related. How

ever, in nine of these males, the cause of death was

attributed to inflammation in various organs, all of

which contained Splendore

- Hoeppli material. Survival

of exposed group s of females decreased with increasing

exposure concentration

, but survival in each exposed

group was not significantly different from that of the

chamber control group.

T ABLE 

### 6

Survival of Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber

Control 125 ppm 250 ppm 500 ppm

Male

Animals initially in study 50 50 50 50

Moribund 24 20 28 35

Natural deaths 3 4 4 2

Animals surviving to study termination 23 26 18 13

Percent probability of survival at end of study a 46 52 36 26

Mean survival (days) b 679 671 654 639

Survival analysis c P=0. 009 P=0. 844N P=0. 262 P=0.0 33



### Female

Animals initially in study 50 50 50 50

Accidental death s d 0 0 0 2

Moribund 13 17 17 23

Natural deaths 3 0 3 1

Animals surviving to study termination 34 33 30 24

Percent probability of survival at end of study 68 66 60 50

Mean survival (days) 702 692 683 637

Survival analysis P=0.0 28 P=0. 858 P=0. 419 P=0. 054

a Kaplan

- Meier determinations

b Mean of all deaths (uncensored, censored, and terminal sacrifice)

c The result of the life table trend test (Tarone, 1975) is in the chamber control column, and the resu lts of the life table pairwise comparisons

(Cox, 1972) with the chamber controls are in the exposed group columns. A lower mortality in an expose d group i s indicated by N

. d Censored from survival analyses

### 1- Bromopropane, NTP TR 564 43

F IGURE 

### 4

Kaplan

- Meier Survival Curves for Rats

Exposed to 1- Bromopropane by Inhalation for 2 Years

### 44 1- Bromopropane, NTP TR 564



## Body Weights

and Clinical and Macroscopic Findings

Mean body weights of exposed groups of male and

female rats were similar to those of the chamber con

trols throughout the study ( Figure 5; Tables 7 and 8 ). In

males, clinical findings that appeared to be related to

1- bromopropane exposure included head mass (0 ppm,

1/50; 125 ppm, 2/50; 250 ppm , 5/50, 500 ppm, 9/50)

and torso/ventral ulcer/ abscess (2/50, 7/50, 6/50, 20/50).

No clinical findings related to exposure to 1

- bromo

propane were observed in females

. In male and female

rats exposed to 1

- bromopropane, there was an exposure

related inc reased incidence of soft, pale

- yellow to green,

variably sized nodules . These lesions were predomi

nantly located in the nose and/or skin, but other sites

included bone, ear, Harderian gland, larynx, lung, mus

cle, peritoneum, preputial gland, and prostat e gland.

The incidence s of these lesions w ere greater in males ( 2

, 3

, 6

, 15 ) than in females ( 1

, 2

, 0

, 7) . In addition, the

number of animals with multiple masses was increased

in the 500 ppm group s . In most cases, these lesions

were microscopically shown to be suppurative inflam

mation , many with Splendore

- Hoeppli material.

### 1- Bromopropane, NTP TR 564 45

F IGURE 

### 5

Growth Curves for Rats Exposed to 1

- Bromopropane

by Inhalation for 2 Years

### 46 1- Bromopropane , NTP TR 564

T ABLE 

### 7

Mean Body Weights and Survival of Male Rats i n the 2

- Year Inhalation Study of 1- Bromopropane

Day s

on

Study Chamber Control 1 25 ppm 25 0 ppm 5 00 ppm

Av. Wt.

(g) No. of

Survivors Av. Wt.

(g) Wt. (% of

controls) No. of

Survivors Av. Wt.

(g) Wt. (% of

controls) No. of

Survivors Av. Wt.

(g) Wt. (% of

controls) No. of

Survivors

1 104 50 103 99 50 103 99 50 104 100 50

9 137 50 136 99 50 138 100 50 138 100 50

16 162 50 163 101 50 165 102 50 166 103 50

23 187 50 191 102 50 194 104 50 194 104 50

30 206 50 213 104 50 217 106 50 218 106 50

37 223 50 232 104 50 238 107 50 242 109 50

44 239 50 251 105 50 257 108 50 260 109 50

51 253 50 268 106 50 273 108 50 275 108 50

58 267 50 283 106 50 287 107 50 290 109 50

65 279 50 297 106 50 301 108 50 304 109 50

72 291 50 309 106 50 314 108 50 318 109 50

79 300 50 320 107 50 323 108 50 328 109 50

86 310 50 330 107 50 331 107 50 337 109 50

114 346 50 366 106 50 366 106 50 370 107 50

142 373 50 390 105 50 391 105 50 394 106 50

170 393 50 413 105 50 411 104 50 410 104 50

198 413 50 432 105 50 428 104 50 426 103 50

226 428 50 448 105 50 446 104 50 438 102 50

254 443 50 462 104 50 459 104 50 448 101 50

282 453 50 473 105 50 467 103 50 454 100 50

310 463 50 485 105 50 479 103 50 462 100 50

338 470 50 493 105 49 484 103 50 467 100 50

366 478 50 508 106 49 498 104 50 474 99 50

394 489 50 517 106 49 507 104 50 483 99 49

422 495 50 523 106 49 517 104 49 486 98 48

450 502 49 530 106 49 525 104 49 493 98 46

478 508 49 540 106 49 532 105 49 503 99 45

506 510 48 546 107 48 536 105 48 505 99 43

534 520 47 551 106 47 545 105 44 513 99 42

562 520 47 551 106 44 552 106 41 517 100 40

590 519 45 555 107 41 550 106 38 522 101 36

618 527 41 557 106 37 557 106 31 521 99 35

646 523 38 564 108 35 556 106 30 517 99 30

660 527 32 568 108 33 556 106 29 520 99 25

674 520 30 565 109 31 550 106 27 520 100 22

688 527 27 566 107 30 546 104 25 514 98 19

702 517 27 557 108 29 544 105 20 512 99 16

716 517 24 551 107 27 541 105 19 503 97 14

Mean for weeks

1

- 13 228 238 10 4 242 10 6 2 4 4 106

14

- 52 420 440 10 5 4 37 104 430 103

53

- 10 3 512 547 10 7 538 105 506 9 9

### 1- Bromopropane, NTP TR 564 47

T ABLE 

### 8

Mean Body Weights and Survival of Fem ale Rats i n the 2

- Year Inhalation Study of 1- Bromopropane

Day s

on

Study Chamber Control 1 25 ppm 25 0 ppm 5 00 ppm

Av. Wt.

(g) No. of

Survivors Av. Wt.

(g) Wt. (% of

controls) No. of

Survivors Av. Wt.

(g) Wt. (% of

controls) No. of

Survivors Av. Wt.

(g) Wt. (% of

controls) No. of

Survivors

1 88 50 87 99 50 88 100 50 88 99 50

9 108 50 108 100 50 109 101 50 106 98 50

16 122 50 122 100 50 125 102 50 121 99 50

23 137 50 137 100 50 140 102 50 135 99 50

30 146 50 147 100 50 151 103 50 147 100 50

37 155 50 157 102 50 161 104 50 157 101 50

44 163 50 166 101 50 170 104 50 166 102 49

51 170 50 172 101 50 177 104 50 170 100 49

58 175 50 177 102 50 183 105 50 176 101 49

65 180 50 183 101 50 188 105 50 183 102 49

72 185 50 187 102 50 194 105 50 188 102 49

79 189 50 192 102 50 198 105 50 194 103 49

86 191 50 196 103 50 202 106 50 198 104 49

114 203 50 210 103 50 217 107 50 214 105 49

142 215 50 220 102 50 227 106 50 224 104 48

170 226 50 229 102 50 237 105 50 232 103 48

198 233 50 238 102 50 246 106 50 241 104 48

226 243 50 248 102 50 256 105 50 247 102 48

254 254 50 258 102 50 266 105 50 257 101 48

282 262 50 265 101 50 272 104 50 263 101 48

310 270 50 274 101 50 281 104 50 271 100 48

338 281 50 283 101 50 290 103 50 279 99 48

366 293 50 297 101 50 300 102 50 289 98 48

394 306 50 309 101 50 313 102 50 301 98 47

422 318 50 318 100 50 321 101 49 304 96 47

450 328 49 326 99 49 333 102 49 320 98 45

478 337 49 334 99 49 343 102 48 328 97 44

506 348 48 344 99 48 352 101 48 333 96 42

534 357 48 351 98 47 357 100 47 340 95 41

562 360 48 353 98 47 362 101 44 346 96 41

590 369 48 362 98 44 370 100 44 354 96 37

618 370 47 361 98 42 370 100 40 350 95 34

646 376 43 363 97 39 376 100 38 353 94 31

660 382 41 366 96 38 375 98 38 355 93 30

674 380 41 367 97 38 376 99 38 356 94 30

688 384 38 371 97 37 379 99 32 362 94 27

702 382 36 371 97 36 374 98 31 358 94 27

716 387 35 373 97 34 374 97 31 357 92 26

Mean for weeks

1

- 13 155 156 101 160 10 4 156 101

14

- 52 243 247 10 2 255 105 248 102

53

- 10 3 355 348 98 3 55 100 338 95

### 48 1- Bromopropane, NTP TR 564



## Pathology and Statistical Analyses

This section describes the statistically significant or

biologically noteworthy changes in the incidences of

malignant mesothelioma and neoplasms or

nonne oplastic lesions of the large intestine, skin,

pancreatic islet s, nose, larynx, trachea, and lung

. Summaries of the inci dences of neoplasms and

nonneoplastic lesions

, statis tical analyses of primary

neoplasms that occurred with an incidence of at least

5% in at least one animal group, and historical

incidences for the neoplasms mentioned in this section

are presented in Appendix A for male rats and

Appendix B for female rats.

Large I ntestine : T he incidence of adenoma of the large

intestine ( colon or rectum ) in 500 ppm females was sig

nificantly greater than that in the chamber controls

( Table s 9, B1, and B2 ). The incidences in the 250 and

500 ppm groups of females exceeded the historical

ranges for controls in inhalation studies and all r ou tes of

administration (Tables 9 and B3 a ). In 250 and 500 ppm

males, the incidence s of adenoma of the large intestine

w ere slightly increased compared to th at in the chamber

controls ; although the inc reas es we re not statistically

significant, the incidence in the 250 ppm group

exceeded the historical control range s for inhalation

studies and all routes (Tables 9 and A 3a ).

L arge intestin e adenomas were polypoid masses that

protruded into the intestinal lumen (Plates 1 and 2)

. The

epithelium lining the glands had fewer goblet cells than

the epithelium from normal glands and was occasionally

thickened by multiple layers of slightly enlarged neo

plastic epithelial cells with basophilic cytoplasm and

enlarged nuc lei. There were variable numbers of mixed

inflammatory cells in the stroma around the glands and

in the stalk. No invasion of the submucosa layer of the

large intestine was observed.

Skin : In male rats, there were exposure concentration

related increased incidences of keratoacanthoma; kera

toacanthoma or squamous cell carcinoma (combined);

and keratoacanthoma, basal cell adenoma, basal cell carcinoma, or squamous cell carcinoma (combined)

(Tables 10, A1, and A2). The incidences of kerato

acanthoma and of keratoacanthoma or squamous cell

carcinoma (combined) in 250 and 500 ppm males were

significantly increased and exceeded the historical con

trol range s for inhalation studies (Tables 10 and A3 b ).

The incidences of keratoacanthoma, basal cell adenoma,

basal cell carcinoma, or squamous cell carcinoma (com

bined) were significantly increased in all exposed

groups of male s and exceeded the historical control

range for inhalation studies. The incidences of basal

cell adenoma in 250 ppm males and of squamo us cell

carcinoma in 500 ppm males were not significantly

increased, but they exceeded the respective historical

control ranges for inhalation studies (Tables 10, A1

, and

A3 b ).

In female rat s , there were increased incidences of squa

mous cell papilloma, keratoacanthoma, basal cell ade

noma, or basal cell carcinoma (combined) in the 500 ppm

group (Tables 10, B1, and B2). Alt hough the increased

incidences were not significant, they exceeded the

respect ive historical control ranges for inhalation studies

(T able s 10 and B3b).

Keratoacanthomas were well

- demarcated, variably

sized, crateriform masses in the dermis composed of

squamous epithelium that formed thick folds. The cen

ter of the mass was filled with abundant keratin. Squa

mous cell carcinomas were characterized as masses

composed of cords of pleomorphic squamous cells infil

trating the dermis and/or subcutis. There were varying

amounts of keratin and fibrous connective tissue, which

separated the cords of squamous cells within the neo

plasms. Basa l cell adenomas were well

- demarcated

masses composed of cords and lobules of basal cells

with areas of sebaceous or squamous differentiation.

Basal cell carcinomas had a similar appearance, but the

cells in the carcinomas were more pleomorphic. The

carci nomas were locally invasive and often contained

areas of necrosis.

### 1- Bromopropane, NTP TR 564 49

T ABLE 

### 9

Incidences of Adenoma of the Large Intestine in Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber

Control 125 ppm 250 ppm 500 ppm

Male

Colon a 50 50 50 50

Adenoma b 0 0 0 1

Rectum 50 50 50 50

Adenoma 0 0 2 0

Colon or Rectum: Adenoma c

Overall r ate d 0/50 (0%) 0/50 (0%) 2/50 (4%) 1/50 (2%)

Adjusted r ate e 0.0% 0.0% 5.3% 2.8%

Terminal r ate f 0/23 (0%) 0/26 (0%) 2/18 (11%) 0/13 (0%)

First i ncidence (days)

— h

— 729 (T) 682

Poly

- 3 test g P=0.197

— i P=0.216 P=0.472



### Fem ale

Colon 50 50 50 50

Adenoma 0 1 1 1

Rectum 50 50 50 50

Adenoma 0 0 1 4

Colon or Rectum: Adenoma j

Overall r ate 0/50 (0%) 1/50 (2%) 2/50 (4%) 5/50 (10%)

Adjusted r ate 0 .0 % 2.3% 4.7% 13.3%

Terminal r ate 0/34 (0%) 1/33 (3%) 1/30 (3%) 4/24 (17%)

First i ncidence (days)

— 730 (T) 607 719

Poly

- 3 test P=0.004 P=0.493 P=0.225 P=0.018

(T) Terminal sacrifice

a Number necropsied

b Number of animals with neoplasm

c Historical incidence for 2

- year inhalation studies with chamber control s (mean ± standard deviation):

0/349; all routes: 2/1,398 (0.1% ± 0.5%), range 0%

- 2%

d Number of animals with neoplasm per nu mber of animals necropsied

e Poly

- 3 estimated neoplasm incidence after adjustment for intercurrent mortality

f Observed incidence at terminal kill

g Beneath the chamber control incidence are the P values associated with the trend test. Beneath the exposed group incidence are the P values

corresponding to pairwise comparisons between the chamber controls and that exposed group. The Poly

- 3 test accounts for differential

mortality in animals that do not reach terminal sacrifice.

h Not applicable; no neopla sm in animal group

i Value of statistic cannot be computed.

j Historical incidence for inhalation studies : 0 /350; all routes 3/1,350 (0.2% ± 0.6%), range 0%

- 2%

### 50 1- Bromopropane , NTP TR 564

T ABLE 

### 10

Incidences of Neoplasms of the Skin in Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber

Control 125 ppm 250 ppm 500 ppm

Male

Number Necropsied 50 50 50 50

Basal Cell Adenoma a ,b 0 1 2 1

Basal Cell Carcinoma c 0 2 1 2

Keratoacanthoma d

Overall r ate e 0/50 (0%) 3 /50 ( 6 %) 6 /50 ( 12 %) 6 /50 ( 12 %)

Adjusted r ate f 0.0% 7.4 % 15.4 % 16.2 %

Terminal r ate g 0/23 (0%) 3 /26 ( 12 %) 2 /18 (11 %) 2 /13 ( 15 %)

First i ncidence (days)

— i 729 (T) 488 562

Poly

- 3 test h P=0. 008 P=0.115 P=0. 012 P=0. 010

Squamous Cell Carcinoma j 1 1 0 2

Keratoacanthoma or Squamous Cell Carcinoma k

Overall r ate 1 /50 ( 2 %) 4 /50 ( 8 %) 6 /50 ( 12 %) 8 /50 ( 16 %)

Adjusted r ate 2.4% 9.8% 15.4 % 21.4 %

Terminal r ate 0 /23 ( 0 %) 3 /26 ( 12 %) 2 /18 ( 11 %) 3 /13 ( 23 %)

First i ncidence (days) 669 711 488 562

Poly

- 3 test P=0. 006 P=0.171 P=0. 044 P=0. 009

Keratoacanthoma, Basal Cell Adenoma, Basal Cell Carcinoma, or Squamous Cell Carcinoma l

Overall r ate 1 /50 ( 2 %) 7 /50 ( 14 %) 9 /50 ( 18 %) 10 /50 ( 20 %)

Adjusted r ate 2.4% 17.0% 22.6 % 26.7 %

Terminal r ate 0 /23 ( 0 %) 5 /26 ( 19 %) 2 /18 ( 11 %) 4 /13 ( 31 %)

First i ncidence (days) 669 585 488 562

Poly

- 3 test P=0. 003 P=0.028 P=0. 006 P=0. 002



### Female

Squamous Cell Papilloma, Keratoacanthoma, Basal Cell Adenoma, or Basal Cell Carcinoma m

Overall r ate 1 /50 ( 2 %) 1 /50 ( 2 %) 1 /50 ( 2 %) 4 /50 ( 8 %)

Adjusted r ate 2.2% 2.3% 2.4 % 10.6 %

Terminal r ate 1/ 34 (3 %) 1/ 33 (3 %) 1/ 30 (3 %) 4/ 24 ( 17 %)

First i ncidence (days) 730 (T) 730 (T) 730 (T) 730 (T)

Poly

- 3 test P=0. 050 P=0.753 P=0. 746 P=0. 128

(T) Terminal sacrifice

a Number of animals with neoplasm

b Historical incidence for 2

- year inhalation studies with chamber control s (mean ± standard deviation): 4/349 (1.2% ± 1.1%), range 0%

- 2%;

all routes: 15/1,398 (1.1% ± 1.4%), range 0%

- 4%

c Historical incidence for inhalation studies: 4 /349 ( 1.1 % ± 2.3 %), range 0% -6 %; all routes: 11 /1,398 (0. 8 % ± 1.5 %), range 0% -6%

d Historical incidence for inhalation studies: 10/349 ( 2.9 % ± 3.7 %), range 0% -8 %; all routes: 66/1,39 8 ( 4.7 % ± 4.2 %), range 0 %- 16 %

e Number of animals with neoplasm per number of animals necropsied

f Poly

- 3 estimated neoplasm incidence after adjustment for intercurrent mortality

g Observed incidence at terminal kill

h Beneath the chamber control incidence are the P values associated with the trend test. Beneath the exposed group incidence are the P values

corresponding to pairwise comparisons between the chamber controls and that exposed group. The Poly

- 3 test accounts for differentia l

mortality in animals that do not reach terminal sacrifice.

i Not applicable; no neoplasm in animal group

j Historical incidence for inhalation studies: 1 /349 (0.3% ± 0.8%), range 0%

- 2%; all routes: 8/1,398 (0.6% ± 0.9%), range 0%

- 2%

k Historical incid ence for inhalation studies: 11/349 (3.2% ± 3.5%), range 0%

- 8%; all routes: 74/1,398 (5.3% ± 4.1%), range 0%

- 16%

l Historical incidence for inhalation studies: 19 /349 (5.5% ± 4.5%), range 0%

- 10%; all routes: 97 /1,398 ( 6.9 % ± 4.9 %), range 0% -20%

m Historical incidence for inhalation studies: 2 /350 ( 0.6 % ± 1. 0 %), range 0%

- 2%; all routes: 16 /1,350 (1. 2 % ± 1.8 %), range 0% -6%

### 1- Bromopropane, NTP TR 564 51

Malignant Mesothelioma : The incidence of malignant

mesothelioma in 500 ppm male rats was significantly

greater than that in the chamber controls (Tables 11, A1,

and A2). The incidence in the 500 ppm group exceeded

the historical ranges for controls in inhalation studies

and all routes (Tables 11 and A3 c ).

Th i s neoplasm w as found in the epididymis in all

affected animals with other tissues variably affected,

particularly the testis, which was affected in all but one

animal. This suggests that the tunica vaginalis, the most common site of origin for mesotheliomas in F344/N rats

( Hall, 1990 )

, wa s the site of origin for all mesotheli

omas in this study. These mesotheliomas were charac

terized by numerous, complex, papillary structures com

posed of pedunculated, fibrovascular stalks covered by

one to several layers of cuboidal to flattened mesothelial

cells (Plates 3 and 4). The stroma was prominent and

often contained clusters of pleomorphic mesothelial

cells that sometimes formed disorganized tubular struc

tures. There was often extensive invas ion of skeletal

muscle and adipose tissue, but minimal invasion of

underlying tissues at other sites.

T ABLE 

### 11

Incidences of Malignant Mesothelioma in Male Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber

Control 125 ppm 250 ppm 500 ppm

Malignant Mesothelioma a

Overall r ate b 0/50 (0%) 2 /50 ( 4 %) 2/50 (4%) 4 /50 ( 8 %)

Adjusted r ate c 0.0% 4.9 % 5. 2% 10 .8%

Terminal r ate d 0/23 (0%) 2 /26 ( 8 %) 0 /18 ( 0 %) 1 /13 ( 8 %)

First i ncidence (days)

— f 729 (T) 536 394

Poly

- 3 test e P=0. 031 P=0.233 P=0.2 20 P=0. 046

(T) Terminal sacrifice

a Historical incidence for 2

- year inhalation studies with chamber control s (mean ± standard deviation): 5 /349 ( 1.4 % ± 2.2 %), range 0% -6 %;

all routes: 35 /1,398 ( 2.5 % ± 2.3 %), range 0% -6%

b Number of animals with neoplasm per number of animals necropsied

c Poly

- 3 estimated neoplasm incidence after adjustment for intercurrent mortality

d Observed incidence at terminal kill

e Beneath the chamber control incidence are the P values associated with the trend test. Beneath the exposed group incidence are the P values

corresponding to pairwise comparisons between the chamber controls and that exposed group. The Poly

- 3 test accounts for differential

mortality in animals that do not reach terminal sacrifice.

f Not applicable; no neoplasm in animal group

### 52 1- Bromopropane, NTP TR 564

P ancreatic Islets : In male rats, the incidences of pan

creatic islet adenoma were significantly increased in all

exposed groups compared to the chamber controls

(Tables 12

, A1, and A2 ). However, the incidences were

with in the historical control ranges for inhalation studies

and for all routes (Table s 12 and A3 d ). The incidences

of pancreatic islet carcinoma were increased in the 125

and 250 ppm groups, but these increas es were not sta

tistically significant. However, the incidence in the

125 ppm group exceeded the historical control ranges

for inhalation studies and for all routes. The incidences

of pancreatic islet adenoma or carcinoma (combined)

were significantly in creased in the 125 and 250 ppm

groups, and the incidence in the 125 ppm group exceeded the historical control ranges for inhalation

studies and for all routes of administration.

Adenomas were usually discrete, well

- circumscribed

masses of islet cells that were 1 mm in diameter or

larger and compressed the surrounding acinar tissue.

Occasionally, adenomas were encapsulated by a thin

band of fibrous connective tissue. In some adenomas,

groups of exocrine pancreatic acini were present within

the adenomas at their periphery. Carcinomas, which

tended to be larger than adenomas, were characterized

by varying degrees of atypia and pleomorphism of the

neoplastic cells, and they typically invaded the sur

rounding fibrous capsule or pancreatic tissue.

T ABLE 

### 12

Incidences of Pancreatic Islet Neoplasms in Male Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber

Control 125 ppm 250 ppm 500 ppm

Adenoma a

Overall r ate b 0/50 (0%) 5 /50 ( 10 %) 4 /50 ( 8 %) 5 /50 ( 10 %)

Adjusted r ate c 0.0% 12.2 % 10.4 % 13.9 %

Terminal r ate d 0/23 (0%) 4 /26 ( 15 %) 1 /18 ( 6 %) 4 /13 ( 31 %)

First i ncidence (days)

— f 666 608 697

Poly

- 3 test e P=0. 043 P=0.029 P=0. 050 P=0. 019

Carcinoma g

Overall r ate 3 /50 ( 6 %) 7 /50 ( 14 %) 5 /50 ( 10 %) 3 /50 ( 6 %)

Adjusted r ate 7.2 % 17.0 % 13.0 % 8 .3 %

Terminal r ate 2 /23 ( 9 %) 3 /26 ( 12 %) 3 /18 ( 17 %) 2 /13 ( 15 %)

First i ncidence (days) 686 687 578 687

Poly

- 3 test P=0. 0516N P=0.149 P=0. 312 P=0. 594

Adenoma or Carcinoma h

Overall r ate 3 /50 ( 6 %) 10 /50 ( 20 %) 9 /50 ( 18 %) 8 /50 ( 16 %)

Adjusted r ate 7.2% 24.2% 23.1 % 22.2 %

Terminal r ate 2 /23 ( 9 %) 5 /26 ( 19 %) 4 /18 ( 22 %) 6 /13 ( 46 %)

First i ncidence (days) 686 666 578 687

Poly

- 3 test P=0. 093 P=0.031 P=0. 043 P=0. 057

a Historical incidence for 2

- year inhalation studies with chamber control s (mean ± standard deviation): 20 /349 ( 5.7 % ± 3.9 %), range 0%

- 12 %;

all routes: 90 /1,39 4 ( 6.5 % ± 3.6 %), range 0%

- 14 %

b Number of animals with neoplasm per number of animals with pancreatic islets examined microscopically

c Poly

- 3 estimated neoplasm incidence after adjustment for intercurrent mortality

d Observed incidence at terminal kill

e Beneath the chamber control inci dence are the P values associated with the trend test. Beneath the exposed group incidence are the P values

corresponding to pairwise comparisons between the chamber controls and that exposed group. The Poly

- 3 test accounts for differential

mortality i n animals that do not reach terminal sacrifice. A negative trend is indicated by N

. f Not applicable; no neoplasm in animal group

g Historical incidence for inhalation studies: 17/349 (4.9% ± 3.3%), range 2%

- 10%; all routes: 29/1,394 (2.1% ± 2.6%), range 0%

- 10%

h Historical incidence for inhalation studies: 37/349 (10.6% ± 4.8%), range 6%

- 18%; all routes: 119/1,394 (8.6% ± 4.0%), range 0%

- 18%

### 1- Bromopropane, NTP TR 564 53

Nose : I ncidence s of suppurative chronic inflammation

in 500 ppm males and fe males were significantly greater

than those in the chamber controls (Table s 13 , A4, and

B4 ). Suppurative chronic inflammation was character

ized by marked numbers of neutrophils with Splendore

Hoeppli material within the nasal cavity lumen, located

primarily in Levels II and III of the nasal sections only

(Plates 5 and 6)

. T he designation of chronic was used to

differentiate suppurative inflammation with Splendore

Hoeppli material from suppurative inflammation with

out Splendore

- Hoeppli material, wh ich was observed in

other organs. Occasionally

, neutrophils in the affected

area would invade the subjacent lamina propria. Appli

cation of special staining for bacteria indicated the pres

ence of Giemsa

- positive and Gram

- negative bacteria

associated wit h the Splendore

- Hoeppli material.

Chronic active inflammation was diagnosed in many

animals of both sexes, including the chamber controls

(Tables 13 , A4, and B4)

. The incidences in all exposed

groups of females were significantly increased com

pared to that in the chamber controls. Chronic active

inflammation was most commonly observed in Level II

of nasal sections, and was characterized by the presence

of lymphocytes and macrophages, with fewer neutro

phils within or around the nasolacrimal duct, or wi thin

the lumina of submucosal glands or the lamina propria

of the septum and turbinates.

The severity of epithelial hyaline droplet accumulation

was increased in all exposed groups of females com

pared to that in the chamber controls (Table 13)

. Hya

line droplet accumulation was characterized by the

presence of intracytoplasmic, eosinophilic globules pri

marily within the olfactory epithelium and to a lesser

extent the respiratory epithelium. In general, this lesion

was seen in many animals in the chambe r control and

exposed groups and it occurred most often in Levels II

and III.

T he incidences of glandular hyperplasia were signifi

cantly increased in all exposed groups of males and

females (T able s 13 , A4, and B4 ). Glandular hyperplasia

occurred most often in Level II of the nasal section, and

was at times associated with inflammation. This lesion

consisted of mild to moderate proliferation of submu

cosal glands

.

The incidences of respiratory epithelial hyperplasia were

significantly increased in 125 and 500 ppm females

(Tables 13 and B4). Respiratory epithelial hyperplasia

was located most often along the septum in Level II and occasionally the ethmoid turbinates in Level III of the

nasal sections. This lesion consisted of thickening of the

respiratory epithelium by increased numbers of tall,

columnar, respiratory epithelial cells.

The incidence of respiratory metaplasia of the olfactory

epithelium was significantly increased in 500 ppm

females (Tables 13 and B4)

. This lesion was primarily

located in Level II at the dorsal meatus and septum but

occasionally in Level III of the nasal sections. In the

affected areas, the olfactory epithelium had been

replaced by ciliated respiratory epithelium.

Larynx: Suppurative chronic inflammation was present

in only one male and three females exposed to 500 ppm

(Tables 13 and B4 )

. T his lesion was not seen in the

chamber controls of either sex

. Suppurative, chronic

inflam mation of the larynx was characterized by marked

numbers of neutrophils, often degenerate, within the

glands and lamina propria with intraglandular Splendore

Hoeppli material ( Plate 7 ).

T he incidences of chronic active inflammation in

250 ppm males and females and 500 ppm females were

significan tly increased (Tables 13 , A4, and B4 ). This

lesion consisted of minimal to moderate numbers of

lymphocytes, macrophages, and neutrophils within the

lamin a propria and glandular lumens.

In females, the incidence of squamous metaplasia in the

500 ppm group was significantly greater than that in the

chamber controls (Table s 1 3 and B4 ). Squamous meta

plasia was characterized by the replacement of normal

respiratory epithelium by squamous epithelial cells.

T

rachea : In females, the incidences of chronic active

inflammation and epithelial hyperplasia were signifi

cantly increased in the 500 ppm group (Tables 13 and

B4 ). Both lesions also occurred in males, but the inci

dences were not significantly increased

. Chr onic active

inflammation was characterized by the presence of

minimal to moderate infiltrates of lymphocytes and

macrophages, with fewer neutrophils in the lamina

propria. Hyperplasia of the epithelium consisted of

increased numbers of respiratory epithelial cells with

densely packed nuclei.

Lun g: S uppurative chronic inflammation occurred in a

few 500 ppm males and females and in on e 250 ppm

male (Table s 13 , A4, and B4 )

. The lesion was mor

phologically similar to that seen in other organs.

### 54 1- Bromopropane , NTP TR 564

T ABLE 

### 13

Incidences of Selected Nonneoplastic Lesions of the Respiratory System in Rats

in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber

Control 125 ppm 250 ppm 500 ppm

Male

Nose a 50 48 48 50

Inflammation, Suppurative, Chronic b 0 1 (4.0) c 2 (4.0) 7** (4.0)

Inflammation, Chronic Active 29 (1.6) 33 (1.4) 34 (1.5) 35 (1.5)

Epithelium, Accumulation,

Hyaline Droplet 44 (1.0) 39 (1.5) 36 (1.3) 44 (1.3)

Glands, Hyperplasia 5 (2.0) 14* (2.0) 14** (2.0) 15** (2.0)

Larynx 50 50 50 50

Inflammation, Suppurative, Chronic 0 0 0 1 (4.0)

Inflammation, Chronic Active 21 (1.4) 28 (1.3) 31* (1.4) 26 (1.3)

Metaplasia, Squamous 4 (1.0) 6 (1.0) 8 (1.1) 5 (1.2)

Trachea 50 50 50 50

Inflammation, Chronic Active 1 (2.0) 1 (1.0) 1 (1.0) 4 (1.5)

Epithelium, Hyperplasia 1 (2.0) 0 0 1 (2.0)

Lung 50 50 50 50

Inflammation, Suppurative, Chronic 0 0 1 (4.0) 3 (4.0)



### Female

Nose 50 50 49 50

Inflammation, Suppurative, Chronic 0 1 (4.0) 3 (4.0) 7** (4.0)

Inflammation, Chronic Active 24 (1.3) 37** (1.5) 37** (1.5) 36** (1.3)

Epithelium, Accumulation,

Hyaline Droplet 48 (1.1) 48 (1.8) 48 (1.7) 47 (1.9)

Glands, Hyperplasia 6 (2.0) 23** (2.0) 28** (2.0) 30** (2.0)

Respiratory Epithelium, Hyperplasia 5 (1.2) 13* (1.3) 9 (1.7) 18** (1.5)

Olfactory Epithelium, Metaplasia,

Respiratory 3 (1.7) 4 (1.8) 6 (1.8) 9* (2.2)

Larynx 50 50 50 50

Inflammation, Suppurative, Chronic 0 0 0 3 (4.0)

Inflammation, Chronic Active 18 (1.1) 25 (1.5) 30** (1.4) 32** (1.5)

Metaplasia, Squamous 3 ( 1.3 ) 2 (1.5) 6 ( 1.3 ) 21** ( 1.7 )

Trachea 50 50 50 50

Inflammation, Chronic Active 0 4 (1.0) 1 (1.0) 6** (1.7)

Epithelium, Hyperplasia 0 0 0 4* (1.8)

Lung 50 50 50 50

Inflammation, Suppurative, Chronic 0 0 0 4* (4.0)

* Significantly different (P ≤ 0.05) from the chamber control group by the Poly

- 3 test

** P ≤ 0.01

a Number of animals with tissue examined microscopically

b N umber of animals with lesion

c Average severity grade of lesions in affected animals: 1=minimal, 2=mild, 3=moderate, 4=marked

### 1- Bromopropane, NTP TR 564 55

Other F indings : S uppurative chronic inflammation with

Splendore

- Hoeppli material was sporadically observed

in several other organs of exposed animals, including

skin [ all exposed groups of males ( with a n exposure

concentration

- related increase in incidence s) , and 125

and 500 ppm females ] , prostate gland ( 500 ppm males),

preputial gland ( 125 ppm males), Harderian gland

(500 ppm males and females), skeletal muscle ( 500 ppm

males), bone (500 ppm males and females ), ear

( 500 ppm females), and peritoneum ( 500 ppm males)

(Table s A4 and B4 ) . The designation of chronic was

given to all diagnoses in which Splendore

- Hoeppli material was present to distinguish it from suppurative

inflammation without Splendore

- Hoeppli material.

Swabs were collected from abscesses on the tail,

Harderian gland, head, and salivary gland of five

exposed rats. The swabs were cultured under aerobic

and anaerobic conditions and bacterial isolates were

identified. No bacterial growth was observed under

anaerobic conditions. Pseudomonas aeruginosa was the

primary isolate in all aerobi c cultures. Splendore

Hoeppli bodies were later observed microscopically in

these lesions

.

### 56 1- Bromopropane, NTP TR 564



# M ICE



# 2-W EEK 

# S TUDY

A ll 2,000 ppm males and two 2,000 ppm females, four

500 ppm males, one 1,000 ppm male, and one

1,000 ppm female died early (Table 14) ; all these early

deaths occurred during the first week of exposure. Final

mean body weights of exposed groups were similar to

those of the chamber controls ; the mean body weight

gain of 1,000 ppm males was significantly decrea sed.

A bnormal breathing, lethargy, and eye discharge were

observed primarily during week 1 and occurred only in

groups exposed to 500 ppm or greater. In the 1,000 ppm males, the absolute and relative heart

weights were significantly less than those of the cham

ber controls while the absolute and relative liver weight s

w ere significantly greater than th ose of the chamber

controls (Table G3 ). A bsolute right kidney weights

were significantly increased in 250 ppm or greater

females, and the relative right k idney weights were sig

nificantly increased in 1,000 and 2,000 ppm females.

Absolute and relative liver weights of 500 ppm or

greater females were significantly greater than those of

the chamber controls. The a bsolute and relative thymus

weights of 1,000 and 2,000 ppm females were signifi

cantly decreased

.

T ABLE 

### 14

Survival and Body Weights of Mice in the 2

- Week Inhalation Study of 1

- Bromopropane

Concentration

(ppm) Survival a 

### Mean Body Weight b 

### (g) Final Weight

Relative to Controls

(%) Initial Final Change

Male

0 5/5 23.9 ± 0.2 28.0 ± 0.4 4.1 ± 0.3

125 5/5 24.2 ± 0.1 28.9 ± 0.5 4.7 ± 0.5 103

250 5/5 23.7 ± 0.3 27.0 ± 0.4 3.3 ± 0.2 96

5 00 1 /5 c 24.0 ± 0. 4 25.8 2.1 92

1,0 00 4 /5 d 23.9 ± 0.2 26.6 ± 0.5 2.6 ± 0.8* 95

2,0 00 0 /5 e 24.1 ± 0.2

— —



### Female

0 5/5 20.0 ± 0.5 22.9 ± 0.2 3.0 ± 0.3

125 5/5 19.4 ± 0.4 22.2 ± 0.5 2.8 ± 0.5 97

250 5/5 20.4 ± 0.2 22.8 ± 0.2 2.5 ± 0.2 100

500 5/5 19.8 ± 0.2 23.4 ± 0.6 3.6 ± 0.5 102

1,000 4/5 f 19.8 ± 0.3 22.8 ± 0.5 3.3 ± 0.6 99

2,0 00 3 /5 d 19.8 ± 0.2 22.7 ± 0.3 3.2 ± 0.2 99

* Significantly different (P ≤ 0.05) from the chamber control group by Williams ’ t est

a Number of animals surviving at 2 weeks /number initially in group

b Weights and weight changes are given as mean ± standard error. Subsequent calculations are based on animals surviving to the end of the

study.

c Day s of death: 4 , 4, 4, 5

d Day of death s : 3

e Day s of death: 2, 2, 2, 2, 3

f Day of death: 5

### 1- Bromopropane, NTP TR 564 57

M icroscopic lesions related to 1

- bromopropane expo

sure occurred in the lung, liver, and nose of males and

females and were primarily seen in mice exposed to

500 ppm or greater (Table 15)

. Bronchiole necrosis

occurred in the lungs of all exposed male and female

m ic e and the severity of th is lesion was increased in the

2,000 ppm groups. The lungs of exposed mice also had

sporadic incidences of regeneration, cytoplasmic vacu

olization, and acute inflammation involving bronchiolar

epithelium. Incidences of centrilobular necrosis

occurred in the liver of most males and females exposed

to 500 ppm or greater. In addition, the incidences of centrilobular chronic inflammation and cytoplasmic

v acuol ization were significantly increased in 1,000 ppm

males and females. Sporadic incidences of nasal lesions

occurred in exposed groups of mice; the no

- effect level

for nose lesions was 250 ppm in males and 500 ppm in

females.

Exposure Concentration Se lection Rationale : Based on

decreased survival and increased liver and kidney

weights in the 2

- week study, 1

- bromopropane exposure

concentrations selected for the 3

- month inhalation study

in mice were 62.5, 125, 250, and 500 ppm.

T ABLE 

### 15

Incidences of Selected Nonneoplastic Lesions in Mice in the 2

- Week Inhalation Study of 1

- Bromopropane

Chamber

Control 12 5 ppm 2 50 ppm 500 ppm 1,000 ppm 2 ,000 ppm

Male

Lung a 5 5 5 5 5 5

Bronchiole, Necrosis b 0 5** (1.0) c 5** (1.0) 5** (2.0) 5** (1.4) 5** (3.4)

Bronchiole, Regeneration 0 5** (1.0) 5** (1.0) 2 (2.0) 4* (1.5) 0

Bronchiole, Vacuolization,

Cytoplasmic 0 0 0 4* (3.0) 1 (3.0) 0

Perivascular, Inflammation, Acute 0 0 0 3 (2.0) 0 5** (1.4)

Liver 5 0 5 5 5 5

Centrilobular, Necrosis 0 0 5** (3.8) 5** (3.2) 5** (2.4)

Centrilobular, Inflammation,

Chronic 0 0 1 (2.0) 4* (3.3) 0

Centrilobular, Vacuolization,

Cytoplasmic 0 0 3 (1.7) 4* (2.0) 0

Nose 5 0 5 5 5 5

Glands, Necrosis 0 0 4* (2.8) 3 (1.7) 5** (2.6)

Inflammation, Suppurative 0 0 0 3 (1.7) 0

Olfactory Epithelium, Atrophy 0 0 0 4* (1.8) 0

Olfactory Epithelium, Necrosis 0 0 4* (2.0) 2 (2.0) 5** (3.8)

Olfactory Epithelium,

Regeneration 0 0 1 (1.0) 4* (2.3) 0

Respiratory Epithelium, Necrosis 0 0 0 1 (3.0) 5** (2.8)

Respiratory Epithelium,

Vacuolization, Cytoplasmic 0 0 4* (3.0) 1 (3.0) 0

### 58 1- Bromopropane , NTP TR 564

T ABLE 

### 15

Incidences of Selected Nonneoplastic Lesions in Mice in the 2

- Week Inhalation Study of 1

- Bromopropane

Chamber

Control 12 5 ppm 2 50 ppm 500 ppm 1,000 ppm 2 ,000 ppm

Female

Lung 5 5 5 5 5 5

Bronchiole, Necrosis 0 5** (1.2) 5** (1.0) 5** (1.0) 5** (1.4) 5** (2.2)

Bronchiole, Regeneration 0 5** (1.2) 5** (1.0) 5** (2.0) 4* (1.8) 3 (2.3)

Bronchiole, Vacuolization,

Cytoplasmic 0 0 0 0 1 (3.0) 0

Perivascular, Inflammation, Acute 0 0 0 0 1 (1.0) 2 (4.0)

Liver 5 0 5 5 5 5

Centrilobular, Necrosis 0 0 3 (1.3) 5** (2.8) 5** (1.8)

Centrilobular, Inflammation,

Chronic 0 0 3 (1.3) 4* (3.0) 3 (1.3)

Centrilobular, Vacuolization,

Cytoplasmic 0 0 3 (1.7) 5** (1.4) 3 (2.0)

Nose 5 0 0 5 5 5

Glands, Necrosis 0 0 1 (3.0) 2 (3.0)

Inflammation, Suppurative 0 0 0 1 (1.0)

Olfactory Epithelium, Atrophy 0 0 4* (1.0) 3 (1.7)

Olfactory Epithelium, Necrosis 0 0 1 (3.0) 2 (4.0)

Olfactory Epithelium,

Regeneration 0 0 4* (1.5) 3 (2.3)

Respiratory Epithelium, Necrosis 0 0 1 (2.0) 2 (2.5)

Respiratory Epithelium,

Vacuolization, Cytoplasmic 0 0 1 (4.0) 0

* Significantly different (P ≤ 0.05) from the chamber control group by the Fisher exact test

** P ≤ 0.0 1

a Number of animals with tissue examined microscopically

b Number of animals with lesion

c Average severity grade of lesions in affected animals: 1=minimal, 2=mild, 3=moderate, 4=marked

### 1- Bromopropane, NTP TR 564 59



# 3-M ONTH 

# S TUDY

One 250 ppm male and four males and five females in

the 500 ppm group s died early (Table 16)

. F inal mean

body weight s and body weight gains of all exposed

groups were similar to those of the chamber control

groups (Table 16 and Figure 6)

. By day 3, l ethargy was

observed in male s and females exposed to 500 ppm, and

abnormal breathing was observed in moribund mice

during week 1.

There were no changes in hematology endpoints that

were considered to be related to 1

- bromopropane expo

sure (Table F2). Absolute right kidney weights of 250 and 500 ppm males,

and the relative right kidney weight of 500 ppm males

were significantly less than those of the chamber controls

(Table G4). Relative liver weights of 250 and 500 ppm

males were significantly increased. The absolute and

relative right kidney, liver, and lung weights of 500 ppm

females and the relative liver weight of 250 ppm females

were significantly greater than those of the chamber

controls

.

T ABLE 

### 16

Survival and Body Weights o f Mice in the 3

- Month Inhalation Study of 1

- Bromopropane

Concentration

(ppm) Survival a 

### Mean Body Weight b 

### ( g ) Final Weight

Relative to Controls

(%) Initial Final Change

Male

0 10 /10 23.6 ± 0.3 39.6 ± 0.8 16.0 ± 0.7

62.5 10/10 23.7 ± 0.2 39.0 ± 0.9 15.3 ± 0.8 98

125 10/10 23.6 ± 0.1 39.6 ± 0.6 15.9 ± 0.6 100

250 9 /10 c 23.6 ± 0.3 37.9 ± 0.7 14.4 ± 0.6 96

500 6 /10 d 23.4 ± 0.3 37.4 ± 1.4 14.2 ± 1.2 95



### Female

0 10/10 19.9 ± 0.2 30.9 ± 0.7 11.0 ± 0.5

62.5 10/10 20.3 ± 0.3 33.0 ± 1.3 12.7 ± 1.1 107

125 10/10 20.4 ± 0.3 31.9 ± 0.8 11.5 ± 0.6 103

250 10/10 19.7 ± 0.3 30.2 ± 0.9 10.4 ± 0.9 98

500 5/10 e 20.2 ± 0.1 31.3 ± 1.4 11.0 ± 1.5 101

a Number of animals surviving at 3 months /number initially in group

b Weights and weight changes are given as mean ± standard error. Differences from the chamber control group are not significant by Dunnett’s

test. Subsequent calculations are based on animals surviving to the end of the study.

c W eek of death: 4

d W eeks of death: 1, 1, 1, 2

e W eek of deaths: 1

### 60 1- Bromopropane , NTP TR 564

F IGURE 

### 6

Growth Curves for Mice Exposed to 1

- Bromopropane

by Inhalation for 3 Months

### 1- Bromopropane, NTP TR 564 61

There were slight increases in the cauda epididymis

weights (9 % and 17 %) and significant decreases in

sperm motility (4 %) in 250 and 500 ppm male mice,

respectively (Table H3). There was also a significant

decrease in the sperm per gram cauda (28%) in 500 ppm

males. Exposed groups of female mice differed sig

nificantly from the chamber controls in the relative

amount of time spent in the various stages of estrous

(Table H4). Female mice in the 500 ppm group spent

significantly more time in extended diest rus than the

chamber controls; 250 ppm females spent significantly

more time in extended estrus than the chamber controls.

The length of the estrous cycle of 500 ppm females was

increased slightly (one half day).

Treatment

- related nasal lesions were seen only in mice

that died early. The i ncidences of cytoplasmic vacuol

ization of the respiratory epithelium were significantly

increased in 500 ppm males and females

, and the inci

dence of necrosis of the respiratory epithelium was sig

nificantly increased in 500 ppm females (Table 17 ).

O lfactory epithelium necrosis and necrosis of the lateral

nasal glands occurred in a few 500 ppm males

. A single

incidence of cytoplasmic vacuolization of the respira

tory epithelium occurred in a male exposed to 250 ppm.

Respiratory epithelial lesions were located in the lateral

walls and turbinates of L evel I. The olfactory necrosis

was located in the dorsal meatus of L evel II and in the

dorsal meatus, septum, and turbinates of L evel III. Vac

uol iz ation consisted of a single, large, clear vacuole dis

placing the cytoplasm. Necrosis was characterized by

cells with increased cytoplasmic eosinophilia with loss

of cellular detail and/or pyknotic or fragmented nuclei.

Necrotic cells were frequently sloughed into the airway s

or missing.

Incidences of c ytoplasmic vacuol iz ation and necrosis of

the respiratory epithelium lining the larynx and trachea

occurred in male and female mice exposed to 500 ppm

that died early and in the one early death male exposed

to 250 ppm ; the inci dences of tracheal cytoplasmic

vacuolization were significantly increased in 500 ppm

males and females (Table 17)

. Vacuol iz ation of the

laryn geal lining epithelium also occurred in one

62.5 ppm female that survived to the end of the study.

Vacuol ization was characterized by cells with a large

expansile clear space occupying the majority of the

cytoplasm. These cells often had a pyknotic nucleus at

the center, suggesting that vacuol iz ation progressed to

necrosis. These cells had often lost their cilia. Hyper

plasia of the tracheal epithelium occurred in two 500 ppm males (one an early death and the other a

terminal animal). Hyperplasia was characterized by

increased cell numbers with loss of the normal organi

zation of the epithelium.

In the lung, i ncidences of cytoplasmic vacuolization of

the bronchiolar epithelium were significantly increased

in 500 ppm males and females (Table 17 ). This lesion

also occurred in the 250 ppm male that died early. The

incidence of necrosis of the bronchiolar epithel ium was

significantly increased in 500 ppm females ; this lesion

also occurred in 500 ppm males and the 250 ppm male

that died early

. Necrosis consisted of epithelial cells

with pyknotic nuclei, sloughed epithelial cells, and areas

of bronchial mucosa devo id of epithelial cells or lined

by attenuated epithelial cells. Cells remaining often

contained colorless vacuoles displacing the cytoplasm.

Regeneration of bronchiolar epithelium occurred in all

groups of mice and was not treatment related

. Regener

ati on was described as flatte ne d and disorganized epi

thelium with variation in nuclear size and location.

Nuclei tended to be apical rather than basilar. This

lesion was graded as minimal in all cases.

In the liver, i ncidences of hepatocellular necrosis a nd

hepatocellular degeneration were significantly increased

in 500 ppm males and females; a single incidence of

cytoplasmic vacuolization occurred in a 500 ppm male

(Table 17 ). These lesions were also seen in the early

death animals in the 500 ppm group s

. N ecrosis often

extended beyond the centrilobular zone and was mas

sive in some animals ; it was characterized by uniform

eosinophilic staining of hepatocytes and loss of cellular

detail. Cytoplasmic degeneration and/or vacuol ization,

which w ere often se en peripheral to the necrosis, con

sisted of swollen and foamy hepatocytes with one large

or many tiny clear vacuoles displacing the cytoplasm

and with centrally located nuclei with condensed chro

matin. Chronic inflammation occurred in 500 ppm ter

minal sacrifice animals of both sexes and in on e male

exposed to 250 ppm , and the incidences of this lesion

were significantly increased in the 500 ppm groups

. The inflammation consisted of lymphocytes, macro

phages, large pigment

- laden macrophages, and occa

si onal multinucleated giant cells. Mineralization

occurred in 500 ppm mice of both sexes t hat survived to

the end of the study, and the incidences of this lesion

were significantly increased in these groups

.

N ecrosis of the adrenal cortex (the zona fasciculata

being most affected with extension into the zona

### 62 1- Bromopropane, NTP TR 564

T ABLE 

### 17

Incidences of Selected Nonneoplastic Lesions in Mice in the 3- Month Inhalation Study of 1

- Bromopropane

Chamber

Control 62.5 ppm 125 ppm 250 ppm 500 ppm

Male

Nose a 10 10 10 10 10

Respiratory Epithelium, Vacuolization

Cytoplasmic b 0 0 0 1 (3.0) c 4* (3.0)

Respiratory Epithelium, Necrosis 0 0 0 0 2 (1.5)

Olfactory Epithelium, Necrosis 0 0 0 0 2 (2.5)

Glands, Necrosis 0 0 0 0 1 (2.0)

Larynx 10 10 10 10 9

Vacuolization Cytoplasmic 0 0 0 1 (1.0) 3 (1.7)

Necrosis 0 0 0 0 2 (1.5)

Trachea 10 10 10 10 10

Vacuolization, Cytoplasmic 0 0 0 1 (3.0) 4* (1.0)

Necrosis 0 0 0 1 (2.0) 0

Hyperplasia 0 0 0 0 2 (1.0)

Lung 10 10 10 10 10

Bronchiole, Vacuolization Cytoplasmic 0 0 0 1 (3.0) 4* (2.5)

Bronchiole, Necrosis 0 0 0 1 (2.0) 3 (2.0)

Bronchiole, Regeneration 3 (1.0) 3 (1.0) 4 (1.0) 7 (1.0) 5 (1.0)

Liver 10 10 10 10 10

Necrosis 0 0 0 0 4* (4.0)

Hepatocyte, Degeneration 0 0 0 0 4* (2.5)

Vacuolization Cytoplasmic 0 0 0 0 1 (1.0)

Inflammation, Chronic 0 0 0 1 (1.0) 6** (1.3)

Mineralization 0 0 0 0 5* (1.8)

### 1- Bromopropane, NTP TR 564 63

T ABLE 

### 17

Incidences of Selected Nonneoplastic Lesions in Mice in the 3- Month Inhalation Study of 1

- Bromopropane

Chamber

Control 62.5 ppm 125 ppm 250 ppm 5 00 ppm

Female

Nose 10 10 10 10 10

Respiratory Epithelium, Vacuolization

Cytoplasmic 0 0 0 0 5* (3.0)

Respiratory Epithelium, Necrosis 0 0 0 0 5* (1.4)

Larynx 10 10 10 10 10

Vacuolization Cytoplasmic 0 1 (1.0) 0 0 1 (1.0)

Necrosis 0 0 0 0 1 (1.0)

Trachea 10 10 10 9 10

Vacuolization Cytoplasmic 0 0 0 0 4* (2.0)

Necrosis 0 0 0 0 2 (2.5)

Lung 10 10 10 10 10

Bronchiole, Vacuolization Cytoplasmic 0 0 0 0 5* (3.2)

Bronchiole, Necrosis 0 0 0 0 5* (2.0)

Bronchiole, Regeneration 1 (1.0) 4 (1.0) 5 (1.0) 5 (1.0) 3 (1.0)

Liver 10 10 10 10 10

Necrosis 0 0 0 0 5* (3.6)

Hepatocyte, Degeneration 0 0 0 0 5 * (2. 2 )

Inflammation, Chronic 0 0 0 0 5* (1.8)

Mineralization 0 0 0 0 4* (1.5)

Adrenal Cortex 10 10 10 10 10

Necrosis 0 0 0 0 5* (3.4)

Inflammation, Chronic 0 0 0 0 2 (2.0)

* Significantly different (P ≤ 0.05) from the chamber control group by the Fisher exact test

** P ≤ 0.0 1

a Number of animals with tissue examined microscopically

b Number of animals with lesion

c Average severity grade of lesions in affected animals: 1=minimal, 2=mild, 3=moderate, 4=marked



### glomerulosa to a lesser degree) occurred in 500 ppm

female mice that died early, and the incidence of this

lesion was significantly increased in this group

(Table 17)

. Chronic inflammation was noted at the cor

ticomedullary junction in the adrenal cortex of two

500 ppm females that survived to the end of the study. Exposure Concentration Selection Rationale : Based on

mortality at the highest exposure concentration, changes

in organ weights, and the incidences of various non

neoplastic lesions in the 3

- month study, 1

- bromo

propane exposure concentrations selected for the 2

- year

inhalation study in mice were 62.5, 125, and 250 ppm.

### 64 1- Bromopropane, NTP TR 564



# 2-Y EAR 

# S TUDY



## Survival

Estimates of 2

- year survival probabilities for male and

female mice are shown in Table 18 and in the Kaplan

Meier survival curves (Figure 7 ). Survival of exposed

groups of mice w as similar to that of the chamber con

trol group s.

. 

## Body Weights and Clinical Findings

Mean body weights of exposed groups of male and

female mice were similar to those of the chamber con

trols throughout the study ( Tables 19 and 20 ; Figure 8 ).

No clinical findings related to exposure to 1

- bromo

propane were observed.

T ABLE 

### 18

Survival of Mice in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber

Control 62.5 ppm 125 ppm 25 0 ppm

Male

Animals initially in study 50 50 50 50

Moribund 6 13 9 9

Natural deaths 7 4 9 5

Animals surviving to study termination 37 33 32 36

Percent probability of survival at end of study a 74 66 64 72

Mean survival (days) b 689 686 703 658

Survival analysis c P=0. 934 P=0. 526 P=0. 486 P=0. 888



### Female

Animals initially in study 50 50 50 50

Moribund 10 7 8 8

Natural deaths 4 3 5 0

Animals surviving to study termination 36 d 40 37 42 d

Percent probability of survival at end of study 72 80 74 82

Mean survival (days) 698 713 699 714

Survival analysis P=0. 363N P=0. 413N P=0. 967N P=0. 298N

a Kaplan

- Meier determinations

b Mean of all deaths (uncensored, censored, and terminal sacrifice)

c The result of the life table trend test (Tarone, 1975) is in the chamber control column, and the resu lts of the life table pairwise comparisons

(Cox, 1972) with the chamber controls are in the exposed group columns. A negative trend or lower mortality in an exposure group i s

indicated by N

. d Includes one animal that died during the last week of the study

### 1- Bromopropane, NTP TR 564 65

F IGURE 

### 7

K aplan

- Meier Survival Curves for Mice

Exposed to 1- Bromopropane by Inhalation for 2 Years

### 66 1- Bromopropane , NTP TR 564

T ABLE 

### 19

Mean Body Weights and Survival of Male Mice i n the 2

- Year Inhalation Study of 1- Bromopropane

Day s

on

Study Chamber Control 62.5 ppm 125 ppm 250 ppm

Av. Wt.

(g) No. of

Survivors Av. Wt.

(g) Wt. (% of

controls) No. of

Survivors Av. Wt.

(g) Wt. (% of

controls) No. of

Survivors Av. Wt.

(g) Wt. (% of

controls) No. of

Survivors

1 23.2 50 23.2 100 50 23.1 100 50 22.8 98 50

12 25.7 50 26.0 101 50 26.0 101 50 25.6 99 50

19 26.8 50 27.2 101 50 27.0 101 50 26.7 100 50

26 27.9 50 28.3 101 50 28.1 101 50 27.6 99 50

33 29.2 50 29.3 100 50 29.0 100 50 28.5 98 50

40 30.2 50 30.0 99 50 29.7 99 50 29.3 97 50

47 31.3 50 31.1 99 50 31.0 99 50 30.1 96 48

54 32.2 50 32.2 100 50 31.6 98 50 31.0 96 48

61 33.3 50 33.3 100 50 32.5 98 50 32.0 96 48

68 34.6 50 34.4 99 50 33.8 98 50 32.9 95 48

75 34.8 50 34.8 100 50 34.3 99 50 33.3 96 48

82 35.2 50 35.5 101 50 34.7 98 50 34.0 96 48

89 36.2 50 36.4 100 50 35.6 98 50 34.7 96 48

117 39.5 50 39.7 100 50 39.0 99 50 38.6 98 48

145 42.1 50 42.1 100 50 40.2 96 50 40.9 97 48

173 44.1 50 44.6 101 50 42.9 97 50 43.1 98 48

201 45.6 50 46.3 102 50 44.6 98 50 44.9 98 48

229 48.4 50 48.6 101 50 47.0 97 50 46.7 97 48

257 49.3 50 49.7 101 50 48.2 98 50 47.9 97 48

285 50.3 50 50.8 101 50 49.5 98 50 49.4 98 48

313 50.6 50 51.1 101 50 49.9 99 50 49.7 98 48

341 51.4 49 51.8 101 50 50.7 99 50 50.5 98 48

369 52.3 49 52.5 101 50 51.4 98 50 51.4 98 47

397 52.0 49 51.9 100 50 51.3 99 50 51.2 99 46

425 52.5 48 52.4 100 48 51.8 99 50 51.7 99 45

453 52.6 48 52.6 100 47 52.2 99 50 52.5 100 45

481 53.0 47 52.3 99 47 51.9 98 50 52.5 99 43

509 53.3 47 52.8 99 47 52.3 98 50 53.2 100 43

537 53.4 47 52.9 99 46 52.2 98 49 53.2 100 43

565 52.8 45 52.6 100 45 51.9 98 49 53.3 101 42

593 53.2 43 53.1 100 43 51.7 97 49 53.5 101 41

621 53.4 41 51.7 97 43 52.2 98 45 53.5 100 40

649 52.6 41 51.0 97 41 51.5 98 42 53.0 101 39

663 52.1 41 51.0 98 38 50.6 97 41 52.6 101 39

677 51.8 41 50.9 98 38 50.8 98 39 52.6 102 39

691 51.3 39 50.4 98 36 50.0 98 37 51.8 101 38

705 50.9 39 50.4 99 35 49.8 98 35 51.3 101 37

719 50.9 37 50.5 99 33 50.4 99 32 51.7 102 36

Mean for weeks

1- 13 30.8 30.9 100 30.5 99 29.9 97

14

- 52 46.8 47.2 10 1 45.8 98 45.7 98

53

- 10 3 52.4 51.8 99 51.4 98 52.4 100

### 1- Bromopropane, NTP TR 564 67

T ABLE 

### 20

Mean Body Weights and Survival of Fem ale Mice i n the 2

- Year Inhalation Study of 1- Bromopropane

Day s

on

Study Chamber Control 62.5 ppm 125 ppm 250 ppm

Av. Wt.

(g) No. of

Survivors Av. Wt.

(g) Wt. (% of

controls) No. of

Survivors Av. Wt.

(g) Wt. (% of

controls) No. of

Survivors Av. Wt.

(g) Wt. (% of

controls) No. of

Survivors

1 19.4 50 19.4 100 50 19.2 99 50 19.1 98 50

12 21.2 50 21.1 100 50 21.3 101 50 21.1 100 50

19 22.3 50 22.3 100 50 22.1 99 50 22.1 99 50

26 23.4 50 23.2 99 50 23.2 99 50 23.3 100 50

33 24.3 50 24.2 99 50 24.2 100 50 24.3 100 50

40 24.8 50 24.7 99 50 25.0 101 50 25.0 101 50

47 26.2 50 25.7 98 50 26.2 100 50 25.8 99 50

54 26.9 50 26.5 99 50 26.4 98 50 26.5 99 50

61 27.3 50 27.0 99 50 27.4 100 50 26.8 98 50

68 27.5 50 27.0 98 50 27.6 100 50 27.2 99 50

75 28.0 50 27.7 99 50 28.2 100 50 27.6 98 50

82 28.7 50 28.4 99 50 28.8 100 50 28.4 99 50

89 29.5 50 28.4 96 50 29.3 100 50 28.5 97 50

117 32.1 50 31.6 99 50 32.6 102 50 31.0 97 50

145 34.6 50 34.3 99 50 34.8 101 50 33.0 95 50

173 37.4 50 37.0 99 50 37.9 101 50 34.8 93 50

201 38.8 50 39.0 100 50 39.9 103 50 36.4 94 50

229 42.1 50 42.1 100 50 43.2 103 50 39.1 93 50

257 44.4 50 44.5 100 50 45.2 102 50 41.0 92 50

285 46.8 50 46.9 100 50 47.5 101 50 43.8 94 50

313 48.2 50 48.1 100 50 48.7 101 50 45.4 94 50

341 49.3 50 49.8 101 50 50.4 102 50 46.6 95 50

369 52.8 50 52.1 99 50 53.2 101 49 49.8 94 50

397 53.6 50 53.6 100 50 54.0 101 49 50.8 95 50

425 55.8 50 55.4 99 50 55.2 99 49 52.7 94 50

453 57.2 50 57.3 100 50 57.8 101 49 55.0 96 50

481 58.6 49 58.5 100 50 58.8 100 49 55.6 95 49

509 60.2 49 59.9 100 49 59.6 99 49 56.9 95 49

537 60.9 49 60.8 100 49 60.3 99 47 58.2 96 49

565 60.4 46 61.3 102 48 60.7 101 47 59.2 98 48

593 61.2 45 62.1 101 47 61.3 100 45 60.1 98 47

621 59.3 43 61.5 104 47 61.0 103 43 60.0 101 47

649 58.2 40 61.6 106 46 59.9 103 42 59.7 103 46

663 56.6 40 60.6 107 45 59.8 106 41 58.8 104 44

677 56.2 38 60.4 107 44 59.5 106 41 58.8 105 44

691 55.8 37 59.5 107 42 58.6 105 40 57.2 103 44

705 55.3 37 59.4 108 42 57.9 105 39 56.5 102 43

719 55.9 36 60.1 108 41 58.0 104 37 55.7 100 42

Mean for weeks

1- 13 25.3 25.0 99 25.3 100 25.1 99

14

- 52 41.5 41.5 100 42.2 102 39.0 94

53

- 10 3 57.4 59.0 103 58.5 102 56.6 99

### 68 1- Bromopropane, NTP TR 564

F IGURE 

### 8

Growth Curves for Mice Exposed to 1

- Bromopropane

by Inhalation for 2 Years

### 1- Bromopropane, NTP TR 564 69



## Pathology and Statistical Analyses

This section describes the statistically significant or

biologically noteworthy changes in the incidences of

neoplasms and /or nonneoplastic lesions of the lung,

nose, larynx, and trachea. Summaries of the incidences

of neoplasms and nonneoplastic lesions , s tatistical anal

yses of primary neoplasms that occurred with an inci

dence of at least 5% in at least one animal group, and

historical incidences for the neoplasms mentioned in

this section are presented in Appendix C for male mice

and Appendix D for female mice

.

Lung : In the females, t here were treatment

- related

increase d incidences of alveolar/bronchiolar adenoma

, alveolar/bronchiolar carcinoma

, and alveolar/bronchio

lar adenoma or carcinoma (combined) . T he incidences

of alveolar/bronchiolar adenoma or carcinoma (com

bined) were significantly increased in all exposed

groups (Table s 21 , D1, and D 2 ). T he incidence of

alveolar/bronchiolar adenoma in 250 ppm females and

the incidenc es of alveolar/bronchiolar carcinoma in 62.5

and 125 ppm females were significantly increased. T he

incidence s of alveolar/bronchiolar adenoma in the

250 ppm group and of alveolar/bronchiolar carcinoma

and alveolar/bronchiolar adenoma or carcinoma (com

bin ed) in all exposed female groups exceed ed the

respect ive historical control range s for inhalation stud

ies (Tables 21 and D3)

.

A lveolar/bronchiolar carcinomas varied from moder

ately differentiated, circumscribed lesions to anaplastic,

poorly circumscri bed, infiltrative lesions. They were

variable in size and shape and their growth patterns

were papillary or solid with regions of squamous meta

plasia, spindle cell differentiation, and necrosis. A lveo

lar/bronchiolar adenomas were usually smaller, well

differentiated, circumscribed lesions that often com

pressed the adjacent parenchyma and m ost were

papillary

.

Cytoplasmic vacuolization of bronchiolar epithelium

occurred in all exposed groups of male and female mice,

and the incidences in all exposed groups of male mice

were significantly increased (Table s 21 , C 3 , and D4 ).

This lesion did not occur in chamber control mice.

Histologically, this lesion was characterized by large,

solitary, clear vacuoles, expanding the cytoplasm of

bronchiolar epithelial cells (Plates 8 through 10)

. Affected epithelial cells often lacked cilia. This lesion

is similar to that described in the 3- month inhalation

study of 1

- bromopropane in mice

. Cytoplasmic vacuolization of bronchiolar epithelium

often accompanied bronchiolar regeneration and was

accompanied by bronchiole necrosis in one 250 ppm

male and one 125 ppm female (Tables 21 , C 3 , and D4).

Bronchiolar regeneration occurred in most exposed

male and female mice, but not in chamber controls

(except for a single chamber control male with minimal

regeneration). The incidences w ere similar among

expose d groups, although slight exposure concentration

related increase s in s everit ies w ere noted. Histologi

cally, bronchiolar regeneration was characterized by

disorganized, elongated, and flattened epithelial cells

lining terminal bronchioles (Plates 8 through 10)

. Regenerative epithelial cells often had increased baso

philia, lacked cilia, had var iable amounts of cytoplasm,

and had increased nuclear to cytoplasmic ratios. Nuclei

of regenerative cells tended to be apical rather than

basilar, and often varied in size and shape

. In the

affected bronchioles there was evidence of cell loss.

Many of the affected bronchioles contained sloughed

epithelial cells and occasional necrotic epithelial cells

characterized by small, pyknotic nuclei and homoge

nous, hypereosinophilic cytoplasm.

Nose: There w ere exposure concentration

- related

increase d incidence s of cytoplasmic vacuolization of

respiratory epithelium in male and female mice ; the inci

dences in all exposed groups of males and in 125 and

250 ppm females were significantly increased (Table s 21

, C3 , and D4 ). This lesion did not occur in chamber con

trol mice. Cytoplasmic vacuolization involved the respi

ra tory epithelium on the lateral surfaces of nasoturbinates

in L evels I and II (Plate 11)

. Histologically, this lesion

was characterized by large, solitary, clear vacuoles

expanding the cytoplasm of respiratory epithelial cells.

Oftentimes, affected epithe lial cells lacked cilia. Occa

sionally, nuclei of affected epithelial cells were pyknotic.

This lesion is similar to that described in the 3- month

inhalation study of 1

- bromopropane in mice

.

T h ere w ere treatment

- related increase d incidence s of

respiratory epithelial hyperplasia in the dorsal mea

tus(es) in L evel I in mice of both sexes (Table s 21 , C 3

, and D4 ) ; the inciden ces in all exposed female groups

and in 62.5 and 250 ppm males were significantly

increased

. Although present in chamber controls, the

incidence and severity of respiratory epithelial hyper

plasia increased with increasing e xposure concentration,

and male mice were more severely affected than female

mice. Histological criteria for respiratory epithelial

### 70 1- Bromopropane , NTP TR 564

T ABLE 

### 21

Incidences of Neoplasms and Nonneoplastic Lesions of the Respiratory System in Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber

Control 62.5 ppm 125 ppm 250 ppm

Male

Lung a 50 50 49 49

Bronchiole , Vacuolization Cytoplasmic b 0 18** (1.7) c 19** (1.7) 17** (1.8)

Bronchiole, Regeneration 1 (1.0) 44** (1.0) 38** ( 1.3 ) 47** ( 1.6 )

Bronchiole, Necrosis 0 0 0 1 (3.0)

Nose 50 50 50 50

Respiratory Epithelium,

Vacuolization Cytoplasmic 0 12** (1.8) 19** (1.9) 20** (2.2)

Respiratory Epithelium, Hyperplasia 16 (1.3) 29** (2.0) 23 (2.0) 26* (2.7)

Olfactory Epithelium, Metaplasia,

Respiratory 0 7** (1.6) 6* (1.3) 3 (1.3)

Olfactory Epithelium, Atrophy 2 (2.0) 4 (2.3) 7 (2.3) 4 (1.8)

Larynx 48 50 48 50

Vacuolization Cytoplasmic 0 5* (1.4) 10** (1.1) 11** (1.6)

Trachea 49 50 47 50

Vacuolization Cytoplasmic 0 15** (1.5) 24** (1.8) 24** (2.3)



### Female

Lung 50 50 50 50

Bronchiole , Vacuolization Cytoplasmic 0 3 (1.7) 4 (1.5) 3 (1.3)

Bronchiole, Regeneration 0 45** (1.2) 43** ( 1.3 ) 49** ( 1.6 )

Bronchiole Necrosis 0 0 1 (3.0) 0

Alveolar/bronchiolar Adenoma, Multiple 0 0 0 2

Alveolar/bronchiolar Adenoma (includes multiple) d

Overall rate e 1/50 (2%) 6/50 (12%) 4/50 (8%) 10/50 (20%)

Adjusted rate f 2.2% 12.8% 8.9% 20.8%

Terminal rate g 1/36 (3%) 6/40 (15%) 4/37 (11%) 7/41 (17%)

First incidence (days) 731 (T) 731 (T) 731 (T) 649

Poly

- 3 test h P=0.007 P=0.064 P=0.181 P=0.006

Alveolar/bronchiolar Carcinoma, Multiple 0 2 1 1

Alveolar/bronchiolar Carcinoma (includes multiple) i

Overall rate 0/50 (0%) 7/50 (14%) 5/50 (10%) 4/50 (8%)

Adjusted rate 0.0% 14.9% 11.1% 8.5%

Terminal rate 0/36 (0%) 6/40 (15%) 5/37 (14%) 4/41 (10%)

First incidence (days)

— j 709 731 (T) 731 (T)

Poly

- 3 test P=0.277 P=0.009 P=0.031 P=0.068

Alveolar/bronchiolar Adenoma or Carcinoma k

Overall rate 1/50 (2%) 9/50 (18%) 8/50 (16%) 14/50 (28%)

Adjusted rate 2.2% 19.2% 17.8% 29.2%

Terminal rate 1/36 (3%) 8/40 (20%) 8/37 (22%) 11/41 (27%)

First incidence (days) 731 (T) 709 731 (T) 649

Poly

- 3 test P<0.001 P=0.010 P=0.016 P<0.001

### 1- Bromopropane, NTP TR 564 71

T ABLE 

### 21

Incidences of Neoplasms and Nonneoplastic Lesions of the Respiratory System in Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber

Control 62.5 ppm 125 ppm 250 ppm

Fem ale (continued)

Nose 50 50 50 50

Respiratory Epithelium,

Vacuolization Cytoplasmic 0 3 (1.3) 5* (1.6) 8** (1.6)

Respiratory Epithelium, Hyperplasia 11 (1.1) 25** (1.4) 28** (1.5) 27** (1.7)

Olfactory Epithelium, Metaplasia,

Respiratory 0 4 (1.0) 5* (1.0) 14** (1.1)

Olfactory Epithelium, Atrophy 0 0 0 6* (1.3)

Larynx 50 50 50 50

Vacuolization Cytoplasmic 0 3 (1.3) 2 (2.0) 2 (2.5)

Trachea 50 49 50 50

Vacuolization Cytoplasmic 0 8** (1.5) 7** (1.9) 4 (2.0)

* Significantly different (P ≤ 0.05) from the chamber control group by the Poly

- 3 test

** P ≤ 0.01

(T) Terminal sacrifice

a Number of animals with tissue examined microscopically

b N umber of animals with lesion

c Average severity grade of lesions in affected animals: 1=minimal, 2=mild, 3=moderate, 4=marked

d Historical incidence for 2

- year inhalation studies with chamber controls (mean ± standard deviation ): 18 /3 50 (5 .1% ± 3.8 %), range 2%-1 2%;

all routes: 73 /1,496 ( 4.9 % ± 3.4 %), range 0% -1 2%

e Number of animals with neoplasm per number of animals with lung examined microscopically

f Poly

- 3 estimated neoplasm incidence after adjustment for intercurrent mo rtality

g Observed incidence at terminal kill

h Beneath the chamber control incidence is the P value associated with the trend test. Beneath the exposed group incidence is the P value

corresponding to pairwise comparisons between the chamber controls and that exposed group. The Poly

- 3 test accounts for differential

mortality in animals that do not reach terminal sacrifice.

i Historical incidence for inhalation studies: 9/350 (2.6% ± 2.8%), range 0%

- 6%; all routes: 59/1,496 (3.9% ± 3.4%), range 0%

- 12%

j Not applicable; no neoplasms in animal group

k Historical incidence for inhalation studies: 27/350 (7.7% ± 3.6%), range 2%

- 12%; all routes: 128/1,496 (8.6% ± 3.7%), range 2%

- 18%



### hyperplasia included thickened epithelium due to

increased cellu larity, submucosal epithelial invagina

tions with intraepithelial crypt

- like structures (pseudo

glands), and/or finger

- like projections of epithelium that

extended into the nasal cavity. Lesions were graded

minimal, mild, moderate, or marked (i.e.

, grades 1, 2, 3,

or 4) depending upon the degree of involvement of one

or both meatus(es). Grade 0 was normal. Grade 1

(minimal ) respiratory epithelial hyperplasia was diag

nosed when the respiratory epithelium displayed focal

or regional mucosal thickening with submucosal epi

thelial invaginations in one or both dorsal meatus(es).

Grade 2 (mild) respiratory epithelial hyperplasia was

diagnosed when at least two thirds of one or both dorsal mea tus(es) displayed respiratory epithelium with sub

mucosal epithelial invaginations and/or epithelial papil

lary projections that together involved < 25% into the

dorsal meatus(es)

. Grade 3 (moderate) respiratory epi

thelial hyperplasia was diagnosed when two thirds to all

of both dorsal meatus(es) displayed respiratory epithe

lium with submucosal epithelial invaginations forming

intraepithelial crypt

- like structures (pseudoglands) and

finger

- like papillary projections of epithelium that

together involved 25 % to 50% into the dorsal

meatus(es). Grade 4 (marked) respirator y epithelial

hyperplasia was diagnosed when approximately all of

both dorsal meatuses had hyperplastic respiratory

epithelium with tortuous, submucosal invaginations

### 72 1- Bromopropane , NTP TR 564

forming intraepithelial crypt

- like structures (pseudo

glands) and finger

- like papillary p rojections of epithe

lium that together involved 50 % to 75% into the dorsal

meatuses.

There w ere treatment

- related increase d incidence s of

respiratory metaplasia of olfactory epithelium in male

mice and exposure concentration

- related increase s in

female mice ; incidences of this lesion were significantly

increased in 62.5 and 125 ppm males and 125 and

250 ppm females (Tables 21 , C 3 , and D 4)

. This lesion

was not observed in chamber control mice. L ocated at

L evels II and/or III of the nasal sections, t he lesion was

characterized as segmental replacement of olfactory epi

thelium by ciliated, pseudostratified epithelium resem

bling respiratory epithelium of the anterior nasal cavity;

this lesion was often associated with submucosal

inflam mation and/or glandu lar hyperplasia. The sever

ity of the l esions w as graded minimal to mild based on

the degree of involvement of the dorsal meatuses and/or

turbinates.

The incidence of olfactory epithelial atrophy was signif

icantly increased in 250 ppm females (Table s 21 and D4 ).

The incidence s of this lesion w ere slightly increased in all

exposed groups of males , but the increases were not

statistically significant (Tables 21 and C 3)

. This lesion

consisted of segmental thinning of the olfactory epi

thelium at L evel II and/or III of the nasal cavity due to

loss of neuronal cells with primarily sustentacular and

basal cells remaining. It was often associated with sub

mucosal inflammation and/or glandular hyperplasia, and

usually was present within the dorsal meatuses

.

L arynx and Trachea : There were increased incidences

of c ytoplasmic vacuolization of respiratory epithelium

in the larynx and the trachea of exposed male s and

female s; the incidences were significantly increased in

both tissues in all exposed groups of males and in the trachea of 62.5 and 125 ppm females (Table s 21 , C 3

, and D4 ). This lesion did not occur in chamber control

mice. Histologically, this lesion was characterized by

large, solitary, clear vacuoles expanding the cyto plasm

of respiratory epithelial cells (Plate 1 2)

. A ffected epi

thelial cells often lacked cilia. Occasionally, nuclei of

affected epithelial cells were pyknotic. This lesion is

similar to that described in the 3- month inhalation study

of 1

- bromopropane in mice.

Other Findings : Hepatocellular carcinomas were not

observed in 250 ppm females (Table D2); the incidence

was significantly less than in the concurrent chamber

controls and lower than the historical control range for

inhalation studies [ 39/350 (11% ± 5 %), range 6%

- 20%].

The incidences of hepatocellular adenomas or carcino

mas (combined) in 62.5 and 250 ppm females were

significantly less than that in the chamber controls. The

incidence of skin sarcoma in 250 ppm females was also

significantly less than that in the chamber controls.



# G ENETIC 

# T OXICOLOGY

1- Bromopropane (doses up to 10,000 μ g/plate) was not

mutagenic in either of two independent bacterial muta

genicity assays, with or without exogenous metabolic

activation (Table E1). Bacterial strains tested included

Salmonella typhimurium strains TA97, TA98, TA100,

and TA1535, as well as Escherichia coli strain

WP2 uvrA / pKM101. In addition to the negative results

in the two bacterial tests, no increases in the frequencies

of micronucleated normoc hromatic erythrocytes were

seen in peripheral blood of male or female B6C3F1

mice exposed for 3 months to 62.5 to 500 ppm 1

- bro

mopropane via inhalation (Table E2). The percentage

of reticulocytes (polychromatic erythrocytes) in the

peripheral blood of ma le and female mice was unaltered

by 1

- bromopropane exposure, suggesting a lack of

chem ical

- associated bone marrow toxicity.P LATE 1

Polypoid adenoma in the large intestine (rectum) of a female F344/N rat

exposed to 500 ppm 1

- bromopropane by inhalation for 2 years. The adenoma

occupies the majority of the intestinal lumen. H&E P LATE 2

Higher magnification of Plate 1. Note the multiple layers of basophilic

neoplastic epithelial cells lining the glands. H&E

P LATE 3

Malignant mesothelioma on the parietal surface of the peritoneal cavity of a

male F344/N rat exposed to 500 ppm 1

- bromopropane by inhalation for

2 years. Note the complex papillary structures. H&E P LATE 4

Higher magnification of Plate 3. Note the cuboidal to flattened neoplastic

mesothelial cells lining the papillary structures and the prominent stroma

containing clusters of neoplastic mesothelial cells. H&EP LATE 5

Suppurative chronic inflammation (arrows) in Level II of the nasal cavity of a

male F344/N rat exposed to 500 ppm 1

- bromopropane by inhalation for

2 years. H&E P LATE 6

Higher magnification of Plate 5. The suppurative chronic inflammation is

composed of abundant eosinophilic necrotic cellular debris, fibrin, and

neutrophils, and contains Splendore

- Hoeppli material (arrow). These lesions

were often bordered by a dense, fibrous capsule. H&E

P LATE 7

Larynx of a male F344/N rat exposed to 500 ppm 1

- bromopropane by

inhalation for 2 years. Note the squamous metaplasia of the lining

epithelium (arrowheads) associated with chronic active inflammation within

the submucosa and suppurative chronic inflammati on within the lumen of the

glands (arrows). H&E P LATE 8

Bronchiolar epithelial regeneration and vacuolization in the lung of a male

B6C3F1 mouse exposed to 250 ppm 1

- bromopropane by inhalation for

2 years. There are sloughed epithelial cells in the lumen of the bronchiole

(arrows) indicative of epithelial cell loss. H&EP LATE 9

Higher magnification of Plate 8 . The regenerated epithelium has prominent,

polyhedral cells and is disorganized with loss of cilia (arrows). There is also

vacuolization of the bronchiolar epithelium. H&E P LATE 10

Bronchiolar epithelial necrosis and cytoplasmic vacuolization in the lung of a

male B6C3F1 mouse exposed to 250 ppm 1

- bromopropane by inhalation for

2 years. Note the abundant necr otic cells sloughing into the bronchiolar lumen

(arrows). H&E

P LATE 11

Cytoplasmic vacuolization of the respiratory epithelium lining the

nasoturbinates (arrows) in Level I of the nasal cavity of a male B6C3F1 mouse

exposed to 250 ppm 1

- bromopropane by inhalation for 2 years. H&E P LATE 12

Vacuolization of the respiratory epithelium (arrows) in the trachea of a male

B6C3F1 mouse exposed to 250 ppm 1- bromopropane by inhalation for

2 years. H&E

### 73



# DISCUSSION AND CONCL USIONS

Occupational exposure to 1

- bromopropane is expected

to increase significantly because of its increasing use as

a presumably less toxic and less flammable replacement

for several high

- production volume chemicals used in

emissive applications such as vapor and immersion

degreasing ope rations and critical cleaning of electron

ics and metals. 1

- Bromopropane has also been pro

posed as a replacement for ozone depleting refrigerants

( Fed. Regist.

, 2000) . 1

- Bromopropane was nominated

for study based on the potential for widespread exposure

to this chemical and a lack of chronic toxicity and carci

nogenicity data. Because previ ous short

- term studies

demonstrated that 1

- bromopropane is a neurotoxicant

and a reproductive toxicant in laboratory animals (Yu

et al ., 1998; Ichihara et al ., 2000 a ), the main focus of

the current studies was to evaluate the chronic toxicity

and carcinoge nicity of 1

- bromopropane.

E xposure to 1

- bromopropane concentrations of 500 ppm

or greater in the 2

- week and 3

- month studies resulted in

early deaths of mice. The liver was identified as a

primary target site in these mice

, and severe centri

lobular necrosis was likely the cause of death. In the

2- week study

, moderate to marked centrilobular

necrosis was present in all male s and most females

exposed to 500 ppm or greater

. In the 3

- month study,

extensive hepatic centrilobular necrosis was present

onl y in 500 ppm mice that died early . Centrilobular

chronic inflammation was present in livers of surviving

500 ppm mice, and no hepatocellular lesions were

observed in mice exposed to lower concentrations

except in one 250 ppm male

. The centrilobular locat ion

of the lesions in the region of the liver that contains the

highest levels of P450 activity suggests that the hepa

totoxicity may be due to a reactive metabolite of

1- bromopropane. In mic e, 1- bromopropane is oxida

tively metabolized in the liver by cy tochrome P450 2E1

(CYP2E1) to reactive metabolites that may be

responsible for the toxicity of 1

- bromopropane (Garner

et al

., 2006, 2007; Liu et al ., 2009 ). 2- Oxo -1- bromo

propane, a highly reactive oxidative metabolite of

1- bromopropane (Robinson et al ., 1989), has been

shown to alkylate proteins and inactivate enzymes

(Beeley and Neurath, 1968; Cohen et al ., 1982 ; Mitchell et al ., 1998 ). Detoxification of 1

- bromopropane metab

olites occurs primarily via glutathione

- S

- transferase

(GST)

– mediated conjuga tion with glutathione ( Garner

et al ., 2006; Liu et al ., 2009 ). Glutathione is important

in protecting tissues from alkylating metabolites.

Marked reduction or depletion of liver glutathione

results in enhanced toxicity (Ketterer et al ., 1983). In

th e current stud ies , mice exposed to concentrations

greater than 250 ppm may have produced levels of

metabolites that exceeded the amount of glutathione

available for conjugation. Oxidative metabolites of

1- bromopropane may also deplete glutathione by

inhib iting enzymes required for glutathione synthesis.

Alternatively, 1

- bromopropane metabolites may inhibit

hepatocellular GST thereby preventing glutathione con

jugation. These mechanisms are supported by recent

studies comparing the susceptibility of three strains of

male inbred mice (BALB/cA

, C57BL/6, DBA/2J) to

1- bromopropane

- mediated hepatotoxicity ( Liu et al

., 2009 ). BALB/c A mice were the most susceptible to

1- bromopropane hepatotoxicity and also had the highest

CYP2E1 activity, the lowe st glutathione levels

, and the

lowest GST activity.

Rats were less sensitive than mice to 1- bromopropane

toxicity in the 2

- week and 3

- month studies. In the

2- week rat study

, there were no treatment

- related deaths

and significant ly lower body weights occurred only in

the 1

, 000 and 2

, 000 ppm groups. Liver weights were

increased in most groups of exposed rats; however,

there was no histopathologic evidence of toxicity. In

the 3

- month study

, a mild cytoplasmic vacuolization

was present in the liver of all 500 and 1

, 000 ppm rats,

and in half of the 250 ppm male rats. Other indications

of mild hepatotoxicity in exposed rats were increased

liver weights and increases in serum sorbit o l dehydro

genase levels

. Qualitatively, rats and mice produce the

same oxidative metabolites of 1

- bromopropane (Garner

et al ., 2007); however, mice were reported to have a

greater capacity to oxidatively metabolize 1

- bromopro

pane than rats (Garner et al ., 2006). This species differ

ence in metabolic capacity may partially explain why

extensive hepatotoxicity and early deaths occurred in

exposed mice and not in rats.

### 74 1- Bromopropane , NTP TR 564

The differential capacity of mice and rats to metabolize

1- bromopropane to reactive metabolites, and the possi

ble re lationship of this capacity to the observed hepato

toxicity in 1

- bromopropane

- exposed mice, may have a

bearing on the design of the bacterial mutagenicity

assays conducted with 1

- bromopropane (Appendix E).

These in vitro assays used rat liver microsome pr ep

arations (S9) to provide exogenous metabolic activation

capability to the assay. Given the reduced levels of

CYP2E1 in rat liver, this standard experimental design

may not have been optimal for assessing the muta

genicity of 1

- bromopropane. However, t he mutagenic

activity reported by Barber et al . (1981) in S almonella

typhi mu rium strains TA100 and TA1535 was seen both

with and without rat liver S9, indicating that 1

- bromo

propane is a direct

- acting mutagen, when testing is con

ducted within a closed sy stem to control for the vola

tility of 1

- bromopropane. Because the NTP bacterial

mutagenicity studies were not conducted within a closed

system, the negative results in the assays may have

resulted from inadequate exposure to the test article.

However, g iven the strength of the responses reported

by Barber et al . (1981), it is curious that no indication of

a mutagenic response was observed in the NTP studies,

even allowing for a reduction in exposure due to vola

til ity. Thus, the effects of volatility an d reduced

CYP2E1 levels in standard rat liver S9 preparations on

the mutagenicity of 1

- bromopropane remain unresolved.

1- Bromopropane also caused lesions in the respiratory

tract of mice, although these lesions were not consid

ered as severe as those in t he liver. In the 2

- week study,

respiratory tract lesions were most severe in mice that

died early after exposure to 500 ppm or greater . Necro

sis and vacuolization of the nasal respiratory and olfac

tory epithelium and prominent necrosis of the bronchio

lar epithelium were present only in early death mice. In

survivors exposed to 500 ppm or greater , lesions con

sisted primarily of regeneration of the olfactory epithe

lium and the bronchiolar epithelium. In the 3

- month

study, mild necrosis of the nasal respiratory and olfac

tory epithelium , mild necrosis of the epithelium lining

the trachea, and minimal necrosis of the epithelium

lining the larynx were observed primarily in 500 ppm

mice that died early. Minimal regeneration of the bron

chiolar epitheliu m was observed in the majority of 250

and 500 ppm mice that surviv ed to the end of the study

. Although t he upper respiratory tract lesions in exposed

mice may be due to a direct irritant effect, metabolism

of 1

- bromopropane to a toxic metabolite by the na sal

epithelium cannot be ruled out.

In the 2

- week and 3

- month studies, r ats were less sus

ceptible than mice to 1- bromopropane

- related lesions in the nasal cavity. In the 2

- week study

, foci of minimal

necrosis of the respiratory epithelium and minimal to

mild suppurative inflammation were present in a few

1, 000 and 2

, 000 ppm male rats. There were no expo

sure

- related nasal lesi ons in rats exposed for 3 months.

Although the mechanism of this species difference is

unclear, it may be associated with diff erences in meta

bolic capability in the nasal epithelium.

The potential immunotoxicity of 1

- bromopropane was

investigated because of earlier reports of immunosuppres

sion following oral exposure to 1

- bromopropane (Lee

et al ., 2007a), and the structurally related chemicals

2- bromopropane (Jeong et al

. , 2002 ), and 1,3

- dibromo

propane (Lee et al ., 2007b). 1

- Bromopropane caused

some evidence of immuno suppression in F 344/N rats and

B6C3F1 mice after inhalation exposure for 4 and

10 weeks (Anderson e t al ., 20 10 )

. Significant decreases

in the IgM response to sheep red blood cells (SRBCs)

were observed in both species after exposure to 1

- bromo

propane (Anderson et al ., 20 10 ). In addition, s ignificant

decreases in the total number of spleen cells and T- cells

were observed in mice after 4 weeks of exposure to

1- bromopropane concentrations ranging from 125 to

1, 000 ppm. Lee et al . (2007a,b) suggested that the immu

notoxicity of 1

- bromopropane and 1,3

- dibromopropane

may result from depletion of glutathione . Immunosup

pression caused by these two chemicals was correlated

with chemical

- induced hepatotoxicity and decreases in

liver and spleen glutathione . Partial depletion of glutathi

one has been shown to affect immune function ( effec tor

phase of cytotoxic T

- cell responses and IL

- 2 dependent

functions) (Dr ö ge et al ., 1994). Although glutathione

levels were not measured in the current studies, it is

possible that the hepatotoxicity and immunotoxicity

observed in mice and rats after inhalation of 1

- bromo

propane may be due to reduced levels of glutathione

.

In the 2- year stud y of 1

- bromopropane in mice

, there

was no reduction in survival and no effect on body

weights of male s or female s. Chronic inhalation of

1- bromopropane resulted in neoplasms in female mice

but not in male mice. There was clear evidence that

1- bromopropane exposure caused increase d in cidences

of alveolar/bronchiolar adenoma or carcinoma (com

bined) in female mice. Because alveolar/bronchiolar

adenomas are known to progress to carcinomas

( Boorman and Eustis, 1990 ), the NTP combines the

incidences of these neoplasms . The incidences of alve

olar/bronchiolar adenoma or carcinoma (combined) in

female mice increased in a n exposure concentration

related manner

, were significantly gr eater than that in

the chamber controls at all exposure con c entrations , and

exceeded the range in historical controls for inhalation

### 1- Bromopropane, NTP TR 564 75

studies

. The incidence of alveolar/bronchiolar adenoma

in female mice exposed to 250 ppm was significantly

increased compared to that in the chamber controls and

exceeded the historical control range s for inhalation

studies and for all routes . The incidences of alveo

lar/bronchiolar carcinoma in 62.5 and 125 ppm female

mice were significantly increased compare d to th e

chamber controls; although not statistically increased,

alveolar/bronchiolar carcinoma occurred in 8% of the

250 ppm female s

. The incidences in all exposed groups

of female mice exceeded the historical control range for

inhalation studies and the incidence in the 62.5 ppm

females exceeded the historical control range from all

routes. I n chamber control female mice a lveo

lar/bronchiolar c arcinoma w as not observed. Chronic

exposure to the structurally related 1,2

- dibromoethane

also caused alveolar/bronchiolar adenomas and carcino

mas in male and female mice and female rats ( NTP,

1982a ).

In the 2- year stud y in rats, survival of 500 ppm males

was reduced ; however, there were no chemical

- related

effects on body weights. E xposure to 1

- bromopropane

for 2 years resulted in neoplasms in rats at various sites.

The treatment

- related increased incidences of adenoma

of the large intestine (colon and rectum ) in female rats

were considered clear evidence of carcinogenic activity

. I ncreased incidence s of a denoma w ere exposure con

centration related, and the incidence in the 500 ppm

group was significantly greater than that in the chamber

controls. Adenomas were found in the large intestine of

two 250 ppm male rats (4%) and in one 500 ppm male

rat (2%). Al though the increased incidences in males

were not statistically significant, the presence of these

neoplasms in females and the low historical occurrence

of th ese neoplasm s suggest that they were treatment

related. Adenomas of the large intestine are rare in

control male and female rats, occurring at a rate of less

th an 0.2%. Although no large intestine carcinomas

were observed in male or female rats in the current

study, adenomas of the large intestine can progress to

carcinomas (Deschner, 1983; Chang, 1 984; Nigro,

1985). Oral treatment of rats with two brominated

methanes ( bromodichloromethane and tribromo

methane ) also resulted in significantly increased inci

dences of adenocarcinoma of the large intestine in both

male and female rats (NTP, 1987, 1989 ). Thus, the

occurrence of adenomas of the large intestine were con

sidered some evidence of carcinogenic activity in male

rats.

C hronic exposure to 1

- bromopropane caused an

increase in the incidences of neoplasms of the skin in

male rats

, and skin neoplasms in female rats may also have been related to exposure

. In males, t he combined

incidences of all neoplas m s of epithe lial origin

increased in a significant exposure concentration

dependent manner. The i ncidences were signifi cantly

increased in all exposed groups of males relative to

chamber controls and exceeded the range in historical

controls from inhalation studies

. The most numerous

neoplasms were identified as keratoacanthoma, a benign

neoplasm that morph ologically resembles squamous ce ll

carcinoma. In males, the incidences of keratoacan

thoma increased with a positive trend ; the incidences in

the 250 and 500 ppm groups were signifi cantly

increased compared to chamber controls and exceeded

the historical control range for inhalation stu dies

. The

increased incidence of keratoacanthoma is a concern

because there is evidence that it can progress to squa

mous cell carcinoma, a highly malignant tumor ( Elwell

et al ., 1990 ). Squamous cell carcinoma is a rare neo

plasm occurring in only 0. 6 % of historical controls from

all studies. Although not statistically significant, the

incidence of squamous cell carcinoma in 500 ppm male

rats was greater than that in the chamber controls and

exceeded the historical control range s for inhalation

studies and for all routes. In females, the incidence of

squamous cell papilloma, keratoacanthoma, basal cell

adenoma, or basal cell carcinoma (combined) in the

500 ppm group exceeded the historical control range for

inhalation studies. Keratoacanthomas occurred in

females, but unlike in the males, the incidences were not

increased compared to the con current chamber controls

or historical controls. No squamous cell carcinomas

were seen in female rats.

The evidence that 1

- br o mopropane exposure was asso

ciated wi th an increased incidence of pancreatic islet

adenoma in male ra ts was equivocal. Althoug h the

increased incidences of pancreatic islet adenoma were

statistically significant at all exposure concentrations,

the incidence in the chamber controls (0/50) was l ess

than the mean in historical controls from inhalation

stu d ies (6%) , and t he incidences in exposed groups did

not exceed the historical control range s of inhalation

studies or all routes. The incidences of pancreatic islet

carcinoma in exposed male ra ts were not significantly

different from that in the chamber controls and were not

considered treatment related.

The evidence for malignant mesothelioma in all organs

was equivocal in male rats. The incidences of malignant

mesothelioma occurred with a po sitive trend, but only the

incidence in the 500 ppm group was significantly

increased. The overall strength of this evidence was

considered equivocal because of the common occurrence

of malignant mesothelioma in male rats in 2

- year NTP

### 76 1- Bromopropane , NTP TR 564

studies. The historical control range for malignant meso

thelioma in male rats for inhalation studies and by all

routes is 0% to 6%. Although the incidence in the

500 ppm group was significantly increased, it was barely

outside the historical control range

.

An unusu al nonneoplastic finding in rats exposed to

1- bromopropane for 2 years was the presence of

Splendore

- Hoeppli reaction material associated with

suppurative inflammation. Lesions with Splendore

Hoeppli material were present primarily in the nose and

skin of exposed male and female rats, although other

sites were affected. The exact composition and mech

anism of formation of Splendore

- Hoeppli material is

unknown, but it is thought to represent deposition of

antigen

- antibody complexes and debris from host

inf lammatory cells (Hussein, 2008). Alternatively, it

may represent glycoproteins, lipid, and calcium derived

from host leukocytes (Bhagavan et al ., 1982). Histolog

ically, Splendore

- Hoeppli material appears as strongly

eosinophilic, amorphous material surrounding or adja

cent to the causative agent, typically fungi, helminthes

or bacteria (Hussein, 2008). The Splendore

- Hoeppli

bodies appear microscopically as distinct radiating, star

like, or club

- shaped configurations. In the United

States, Splendore

- Hoe ppli bodies are often seen in asso

ciation with botryomycotic infections caused by

Proteus

, Escherichia coli or Pseudomonas aeruginosa

( Wenig et al ., 1996 ). In this study, cu l tures from four of

five 1- bromopropane

- exposed rats with this lesion were

positive for P. aeruginosa

. It is not clear why lesions

with Splendore

- Hoeppli bodies were only present in

rats, since both rats and mice were immunosup p ressed

after 1

- bromopropane exposure. Species differences in

the p resence of opportunistic bacteria, or differences in

innate resistance to infection

, may have played a role in

this difference.

The presence of lesions with Splendore

- Hoeppli bodies in

rats was considered to be treatment related. The inci

dence of lesion s with Splendore

- Hoeppli bodies increased

with 1

- bromopropane exposure concentration, and was

considerably higher in males (34%) and females (28%)

exposed to 500 ppm. Lesions with Splendore

- Hoeppli

bodies were not present in chamber control rats.

Althoug h the mechanism is not clear, immunosuppres

sion in 1

- bromopropane

- exposed rats may have contri

buted to the development of Splendore

- Hoeppli bodies.

The development of botryomycosis and formation of

Splendore

- Hoeppli bodies in humans may be facilitated

b y immunosuppression as it has been associated with

diabetes, immunoglobulin deficiency, cutaneous anergy, co rticosteroid treatment, and AIDS (Schlossberg et al

., 1980 ; Brunken et al ., 1983; Patterson et al ., 1987 ).

Because adequate data were available demonstrating the

developmental and reproductive toxicity of 1

- bromo

propane, only sperm motility and vaginal cytology were

evaluated in the current studies. Based upon the effects

on sperm count and motility and estrous cyclicity seen

in the 3- month stud ies, we conclude that 1

- bromo

propane has the potential to produce adverse effects on

fe r tility and reproductive performance in rodent studies

involving similar exposures

. The NTP Center for the

Evaluation of Risks to Human Reproduction (CERHR)

conducted an extensive evaluation on the potential

effects of 1

- bromopropane on human reproduction and

development ( NTP , 2003). The CERHR concluded that

based upon the available data, inhaled 1

- bromopropane

causes prenatal developmental toxicity and reproductive

to xicity in male and female rats; however, available

human data were insufficient to draw conclusions on the

potential for reproductive or developmental toxicity.

The overall NTP conclusion was that “there is serious

concern for reproductive and developmental effects of

1- bromopropane at the upper end of the human occupa

tional exposure range (18 to 381 ppm).” “Serious con

cern” is the highest level of NTP conclusion regarding

the possibilities that human development and reproduc

tion might be adversely aff ected.



# C ONCLUSIONS

Under the conditions of these 2

- year inhalation studies,

there was some evidence of carcinogenic activity * of

1- bromopropane in male F344/N rats based on the

occurrence of rare adenomas of the large intestine and

increased incidences of epithelial neoplasms of the skin

(keratoacanthoma, squamous cell carcinoma, and basal

cell neoplasms). Increased incidences of malignant

mesothelioma and pancreatic islet adenoma and

carcinoma (combined) may also have been related to

1- bromopropane exposu re. There was clear evidence of

carcinogenic activity of 1

- bromopropane in female

F344/N rats based on increased incidences of adenoma

of the large intestine. Increased incidences of skin

neoplasms may also have been related to 1

- bromopro

pane exposure

. There was no evidence of carcinogenic

activity of 1

- bromopropane in male B6C3F1 mice

exposed to concentrations of 62.5, 125, or 250 ppm

1- bromopropane. There was clear evidence of car

cinogenic activity of 1

- bromopropane in female

B6C3F1 mice based on in creased incidences of alveo

lar/bronchiolar neoplasms

.

### 1- Bromopropane, NTP TR 564 77

Exposure to 1

- bromopropane resulted in increased

incidences of nonneoplastic lesions in the nose of rats

and mice, the larynx of rats and male mice, and the

trachea and lung of female ra ts and male and female mice. Suppurative inflammatory lesions with

Splendore

- Hoeppli material were present primarily in

the nose and skin of male and female rats exposed to

1- bromopropane

.

* Explanation of Levels of Evidence of Carcinogenic Activity is on page 13 . A summary of the Technical Reports Review Subcommittee

comments and the public discussion on this Technical Report appears on page 15.

### 78 1- Bromopropane , NTP TR 564

### 79



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Margolin, B.H., and Tennant, R.W. (1990). Evaluation

of four in vitro genetic toxicity tests for predicting

rodent carcinogenicity: Confirmation of earlier results

with 41 additional chemicals. Environ. Mol. Mutagen.

16 (Suppl. 1 8), 1

- 14.

Zeiger, E., Anderson, B., Haworth, S., Lawlor, T., and

Mortelmans, K. (1992). Salmonella mutagenicity tests:

V. Results from the testing of 311 chemicals. Environ.

Mol. Mutagen. 19 (Suppl. 21), 2

- 141.

### 85



# APPENDIX A

SUMMARY OF LESIONS I N MALE RATS

IN THE 2

- YEAR INHALATION STUDY

OF 1- BROMOPROPANE

T ABLE 

### A1 Summary of the Incidence of Neoplasms in Male Rats

in the 2

- Year Inhalation Study of 1- Bromopropane

......................................................................... 86

T ABLE 

### A2 Statistical Analysis of Primary Neoplasms in Male Rats

in the 2

- Year Inhalation Study of 1- Bromopropane

........................................................................ 90

T ABLE 

### A3 a Historical Incidence of Large Intestine (Colon or Rectum) Neoplasms

i n Control Male F344/N Rats

.............................................................................................................. 94

T ABLE 

### A3 b Historical Incidence of Skin Neoplasms in Control Male F344/N Rats

........................................... 95

T ABLE 

### A3 c Historical Incidence of Malignant Mesothelioma i n Control Male F344/N Rats

............................ 96

T ABLE 

### A3 d Historical Incidence of Pancreatic Islet Neoplasms i n Control Male F344/N Rats

........................ 96

T ABLE 

### A4 Summary of the Incidence of Nonneoplastic Lesions in Male Rats

in the 2

- Year Inhalation Study of 1- Bromopropane

........................................................................ 97

### 86 1- Bromopropane, NTP TR 564

T ABLE 

### A1

Summary of the Incidence of Neoplasms in Male Rats in the 2

- Year Inhalation Study of 1- Bromopropane a



### Chamber Control 125 ppm 250 ppm 500 ppm

Disposition Summary

Animals initially in study 50 50 50 50

Early deaths

Moribund 24 20 28 35

Natural deaths 3 4 4 2

Survivors

Terminal sacrifice 23 26 18 13

Animals examined microscopically 50 50 50 50



### Alimentary System

Esophagus (50) (50) (50) (50)

Intestine l arge, c ecum (47) (47) (47) (48)

Intestine l arge, c olon (48) (48) (48) (48)

Adenoma 1 (2%)

Intestine l arge, r ectum (48) (48) (48) (49)

Adenoma 2 (4%)

Carcinoid t umor m alignant 1 (2%)

Polyp a denomatous 1 (2%)

Intestine s mall, d uodenum (48) (48) (48) (48)

Intestine s mall, i leum (47) (47) (46) (48)

Intestine s mall, j ejunum (47) (47) (46) (48)

Adenoma 1 (2%)

Liver (50) (50) (50) (50)

Hepatocellular c arcinoma 2 (4%) 1 (2%)

Mesentery (13) (13) (8) (19)

Sarcoma 1 (13%)

Oral m ucosa (3) (0) (2) (4)

Squamous c ell c arcinoma 1 (33%) 1 (50%) 2 (50%)

Pancreas (50) (50) (50) (50)

Salivary g lands (50) (50) (50) (50)

Stomach, f orestomach (50) (50) (50) (50)

Stomach, g landular (50) (50) (50) (50)

Tongue (0) (0) (1) (1)



### Cardiovascular System

Heart (50) (50) (50) (50)



### Endocrine System

Adrenal c ortex (50) (50) (50) (50)

Adenoma 1 (2%) 1 (2%)

Adrenal m edulla (50) (50) (50) (50)

Pheochromocytoma b enign 8 (16%) 8 (16%) 5 (10%) 7 (14%)

Pheochromocytoma c omplex 1 (2%) 1 (2%)

Pheochromocytoma m alignant 1 (2%) 1 (2%) 1 (2%)

Bilateral, p heochromocytoma b enign 2 (4%) 1 (2%) 2 (4%) 1 (2%)

Islets, p ancreatic (50) (50) (50) (50)

Adenoma 5 (10%) 4 (8%) 5 (10%)

Carcinoma 3 (6%) 7 (14%) 5 (10%) 3 (6%)

Parathyroid g land (46) (48) (47) (46)

Pituitary g land (50) (50) (50) (49)

Pars d istalis, a denoma 37 (74%) 36 (72%) 33 (66%) 31 (63%)

### 1- Bromopropane, NTP TR 564 87

T ABLE 

### A1

Summary of the Incidence of Neoplasms in Male Rats in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Endocrine System (continued)

Thyroid g land (50) (50) (50) (49)

C

- cell, a denoma 5 (10%) 4 (8%) 3 (6%) 3 (6%)

C

- cell, c arcinoma 4 (8%) 3 (6%) 2 (4%)

Follicular c ell, a denoma 2 (4%)

Follicular c ell, c arcinoma 2 (4%) 2 (4%) 1 (2%)



### General Body System

Peritoneum (0) (0) (0) (2)



### Genital System

Coagulating g land (3) (0) (1) (2)

Epididymis (50) (50) (50) (50)

Penis (0) (3) (1) (1)

Preputial g land (50) (50) (49) (50)

Adenoma 4 (8%) 1 (2%) 1 (2%) 1 (2%)

Carcinoma 2 (4%) 2 (4%) 3 (6%)

Prostate (50) (50) (50) (50)

Adenoma 1 (2%)

Seminal v esicle (50) (49) (49) (50)

Testes (50) (50) (50) (50)

Bilateral, i nterstitial c ell, a denoma 22 (44%) 20 (40%) 16 (32%) 17 (34%)

Interstitial c ell, a denoma 12 (24%) 18 (36%) 16 (32%) 12 (24%)



### Hematopoietic System

Bone m arrow (50) (50) (50) (50)

Lymph n ode (5) (5) (5) (4)

Deep c ervical, c arcinoma, m etastatic,

thyroid gland 1 (20%)

Lymph n ode, b ronchial (3) (6) (5) (8)

Lymph n ode, m andibular (3) (1) (0) (6)

Lymph n ode, m ediastinal (30) (25) (23) (32)

Carcinoma, m etastatic, t hyroid g land 1 (3%)

Lymph n ode, m esenteric (50) (50) (50) (50)

Spleen (50) (50) (49) (50)

Thymus (43) (41) (47) (46)



### Integumentary System

Mammary g land (40) (36) (39) (36)

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

Skin (50) (50) (50) (50)

Basal c ell a denoma 1 (2%) 2 (4%) 1 (2%)

Basal c ell c arcinoma 2 (4%) 1 (2%) 2 (4%)

Fibrous h istiocytoma 1 (2%)

Hemangiosarcoma 1 (2%)

Keratoacanthoma 3 (6%) 4 (8%) 5 (10%)

Keratoacanthoma, m ultiple 2 (4%) 1 (2%)

Squamous c ell c arcinoma 1 (2%) 1 (2%) 2 (4%)

Pinna, n eural c rest t umor 1 (2%)

Sebaceous g land, a denoma 2 (4%) 1 (2%)

Sebaceous g land, c arcinoma 1 (2%)

Subcutaneous t issue, f ibroma 3 (6%) 5 (10%) 1 (2%)

Subcutaneous t issue, l ipoma 2 (4%) 1 (2%) 1 (2%)

### 88 1- Bromopropane, NTP TR 564

T ABLE 

### A1

Summary of the Incidence of Neoplasms in Male Rats in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Musculoskeletal System

Bone (50) (50) (50) (50)

Chordoma 1 (2%)

Osteosarcoma 1 (2%)

Skeletal m uscle (1) (1) (3) (3)

Chordoma, m etastatic, b one 1 (100%)



### Nervous System

Brain (50) (50) (50) (50)

Oligodendroglioma m alignant 1 (2%)

Osteosarcoma 1 (2%)

Spinal c ord (1) (1) (2) (1)



### Respiratory System

Larynx (50) (50) (50) (50)

Carcinoma, m etastatic, t hyroid g land 1 (2%)

Lung (50) (50) (50) (50)

Alveolar/ b ronchiolar a denoma 2 (4%) 3 (6%) 1 (2%) 5 (10%)

Alveolar/ b ronchiolar a denoma, m ultiple 1 (2%)

Alveolar/ b ronchiolar c arcinoma 1 (2%) 1 (2%) 2 (4%) 1 (2%)

Carcinoma, m etastatic, p reputial g land 1 (2%)

Carcinoma, m etastatic, t hyroid g land 1 (2%)

Chordoma, m etastatic, b one 1 (2%)

Nose (50) (48) (48) (50)

Trachea (50) (50) (50) (50)

Carcinoma, m etastatic, t hyroid g land 1 (2%)



### Special Senses System

Eye (49) (49) (50) (50)

Melanoma m alignant 1 (2%)

Harderian g land (50) (50) (50) (50)

Lacrimal g land (1) (0) (0) (0)

Zymbal ’ s g land (0) (1) (2) (2)

Adenoma 1 (50%)

Carcinoma 1 (100%) 2 (100%) 1 (50%)



### Urinary System

Kidney (50) (50) (50) (49)

Liposarcoma 1 (2%)

Stromal n ephroma 1 (2%)

Cortex, r enal t ubule, a denoma 1 (2%)

Pelvis, p apilloma 1 (2%)

Urinary b ladder (50) (50) (50) (50)

Transitional e pithelium, c arcinoma 1 (2%)



### Systemic Lesions

Multiple o rgans b (50) (50) (50) (50)

Histiocytic s arcoma 1 (2%) 2 (4%)

Leukemia m ononuclear 29 (58%) 22 (44%) 16 (32%) 24 (48%)

Lymphoma m alignant 1 (2%)

Mesothelioma m alignant 2 (4%) 2 (4%) 4 (8%)

### 1- Bromopropane, NTP TR 564 89

T ABLE 

### A1

Summary of the Incidence of Neoplasms in Male Rats in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Neoplasm Summary

Total a nimals with p rimary n eoplasms c 50 50 48 49

Total p rimary n eoplasms 150 160 139 143

Total a nimals with b enign n eoplasms 48 49 47 48

Total b enign n eoplasms 102 110 98 96

Total a nimals with m alignant neoplasms 38 33 32 32

Total m alignant n eoplasms 47 50 41 47

Total a nimals with m etastatic neoplasms 3 1

Total m etastatic n eoplasms 4 4

Total a nimals with uncertain n eoplasms

- benign or malignant 1

Total u ncertain n eoplasms 1

a Number of animals examined microscopically at the site and the number of animals with neoplasm

b Number of animals with any tissue examined microscopically

c Primary neoplasms: all neoplasms except metastatic neoplasms

### 90 1- Bromopropane, NTP TR 564

T ABLE 

### A2

Statistical Analysis of Primary Neoplasms in Male Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Adrenal Medulla: Benign Pheochromocytoma

Overall rate a 10/50 (20%) 9/50 (18%) 7/50 (14%) 8/50 (16%)

Adjusted rate b 23.9% 21.7% 18.1% 21.6%

Terminal rate c 8/23 (35%) 7/26 (27%) 3/18 (17%) 3/13 (23%)

First incidence (days) 660 585 618 453

Poly

- 3 test d P=0.432N P=0.506N P=0.354N P=0.509N



### Adrenal Medulla: Benign, Malignant, or Complex Pheochromocytoma

Overall rate 11/50 (22%) 10/50 (20%) 8/50 (16%) 10/50 (20%)

Adjusted rate 25.8% 24.1% 20.6% 26.7%

Terminal rate 8/23 (35%) 8/26 (31%) 4/18 (22%) 4/13 (31%)

First incidence (days) 429 585 618 453

Poly

- 3 test P=0.525 P=0.526N P=0.385N P=0.566



### Lung: Alveolar/ b ronchiolar Adenoma

Overall rate 2/50 (4%) 4/50 (8%) 1/50 (2%) 5/50 (10%)

Adjusted rate 4.8% 9.8% 2.7% 13.7%

Terminal rate 2/23 (9%) 3/26 (12%) 1/18 (6%) 3/13 (23%)

First incidence (days) 729 (T) 648 729 (T) 575

Poly

- 3 test P=0.162 P=0.331 P=0.533N P=0.165



### Lung: Alveolar/ b ronchiolar Adenoma or Carcinoma

Overall rate 3/50 (6%) 5/50 (10%) 3/50 (6%) 6/50 (12%)

Adjusted rate 7.2% 12.2% 8 .0 % 16.4%

Terminal rate 3/23 (13%) 4/26 (15%) 2/18 (11%) 3/13 (23%)

First incidence (days) 729 (T) 648 725 575

Poly

- 3 test P=0.169 P=0.349 P=0.620 P=0.181



### Pancreatic Islets : Adenoma

Overall rate 0/50 (0%) 5/50 (10%) 4/50 (8%) 5/50 (10%)

Adjusted rate 0 .0 % 12.2% 10.4% 13.9%

Terminal rate 0/23 (0%) 4/26 (15%) 1/18 (6%) 4/13 (31%)

First incidence (days)

— e 666 608 697

Poly

- 3 test P=0.043 P=0.029 P=0.050 P=0.019



### Pancreatic Islets: Carcinoma

Overall rate 3/50 (6%) 7/50 (14%) 5/50 (10%) 3/50 (6%)

Adjusted rate 7.2% 17 .0 % 13 .0 % 8.3%

Terminal rate 2/23 (9%) 3/26 (12%) 3/18 (17%) 2/13 (15%)

First incidence (days) 686 687 578 687

Poly

- 3 test P=0.516N P=0.149 P=0.312 P=0.594



### Pancreatic Islets: Adenoma or Carcinoma

Overall rate 3/50 (6%) 10/50 (20%) 9/50 (18%) 8/50 (16%)

Adjusted rate 7.2% 24.2% 23.1% 22.2%

Terminal rate 2/23 (9%) 5/26 (19%) 4/18 (22%) 6/13 (46%)

First incidence (days) 686 666 578 687

Poly

- 3 test P=0.093 P=0.031 P=0.043 P=0.057



### Pituitary Gland ( Pars Distalis ): Adenoma

Overall rate 37/50 (74%) 36/50 (72%) 33/50 (66%) 31/49 (63%)

Adjusted rate 79.8% 76 .0 % 73.4% 71.3%

Terminal rate 19/23 (83%) 20/26 (77%) 12/18 (67%) 9/13 (69%)

First incidence (days) 534 505 514 394

Poly

- 3 test P=0.189N P=0.421N P=0.307N P=0.226N

### 1- Bromopropane, NTP TR 564 91

T ABLE 

### A2

Statistical Analysis of Primary Neoplasms in Male Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Preputial Gland: Adenoma

Overall rate 4/50 (8%) 1/50 (2%) 1/49 (2%) 1/50 (2%)

Adjusted rate 9.6% 2.5% 2.7% 2.8%

Terminal rate 3/23 (13%) 0/26 (0%) 0/18 (0%) 0/13 (0%)

First incidence (days) 669 711 688 642

Poly

- 3 test P=0.152N P=0.184N P=0.212N P=0.224N



### Preputial Gland: Carcinoma

Overall rate 2/50 (4%) 2/50 (4%) 3/49 (6%) 0/50 (0%)

Adjusted rate 4.8% 4.9% 7.9% 0 .0 %

Terminal rate 1/23 (4%) 1/26 (4%) 1/18 (6%) 0/13 (0%)

First incidence (days) 660 660 575

— Poly

- 3 test P=0.257N P=0.688 P=0.458 P=0.272N



### Preputial Gland: Adenoma or Carcinoma

Overall rate 6/50 (12%) 3/50 (6%) 4/49 (8%) 1/50 (2%)

Adjusted rate 14.3% 7.3% 10.5% 2.8%

Terminal rate 4/23 (17%) 1/26 (4%) 1/18 (6%) 0/13 (0%)

First incidence (days) 660 660 575 642

Poly

- 3 test P=0.080N P=0.251N P=0.428N P=0.082N



### Skin: Keratoacanthoma

Overall rate 0/50 (0%) 3/50 (6%) 6/50 (12%) 6/50 (12%)

Adjusted rate 0 .0 % 7.4% 15.4% 16.2%

Terminal rate 0/23 (0%) 3/26 (12%) 2/18 (11%) 2/13 (15%)

First incidence (days)

— 729 (T) 488 562

Poly

- 3 test P=0.008 P=0.115 P=0.012 P=0.010



### Skin: Basal Cell Adenoma or Basal Cell Carcinoma

Overall rate 0/50 (0%) 3/50 (6%) 3/50 (6%) 3/50 (6%)

Adjusted rate 0 .0 % 7.3% 7.8% 8.3%

Terminal rate 0/23 (0%) 2/26 (8%) 0/18 (0%) 2/13 (15%)

First incidence (days)

— 585 618 687

Poly

- 3 test P=0.111 P=0.117 P=0.105 P=0.094



### Skin: Keratoacanthoma or Squamous Cell Carcinoma

Overall rate 1/50 (2%) 4/50 (8%) 6/50 (12%) 8/50 (16%)

Adjusted rate 2.4% 9.8% 15.4% 21.4%

Terminal rate 0/23 (0%) 3/26 (12%) 2/18 (11%) 3/13 (23%)

First incidence (days) 669 711 488 562

Poly

- 3 test P=0.006 P=0.171 P=0.044 P=0.009



### Skin: Basal Cell Carcinoma or Squamous Cell Carcinoma

Overall rate 1/50 (2%) 3/50 (6%) 1/50 (2%) 4/50 (8%)

Adjusted rate 2.4% 7.4% 2.6% 11 .0 %

Terminal rate 0/23 (0%) 2/26 (8%) 0/18 (0%) 2/13 (15%)

First incidence (days) 669 711 697 659

Poly

- 3 test P=0.124 P=0.297 P=0.739 P=0.138



### Skin: Keratoacanthoma

, Basal Cell Adenoma, Basal Cell Carcinoma, or Squamous Cell Carcinoma

Overall rate 1/50 (2%) 7/50 (14%) 9/50 (18%) 10/50 (20%)

Adjusted rate 2.4% 17 .0 % 22.6% 26.7%

Terminal rate 0/23 (0%) 5/26 (19%) 2/18 (11%) 4/13 (31%)

First incidence (days) 669 585 488 562

Poly

- 3 test P=0.003 P=0.028 P=0.006 P=0.002

### 92 1- Bromopropane, NTP TR 564

T ABLE 

### A2

Statistical Analysis of Primary Neoplasms in Male Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Skin (Subcutaneous Tissue): Fibroma

Overall rate 3/50 (6%) 5/50 (10%) 1/50 (2%) 0/50 (0%)

Adjusted rate 7.1% 12 .0 % 2.6% 0 .0 %

Terminal rate 1/23 (4%) 2/26 (8%) 0/18 (0%) 0/13 (0%)

First incidence (days) 646 560 690

— Poly

- 3 test P=0.053N P=0.349 P=0.342N P=0.149N



### Skin : Fibroma or Fibrous Histiocytoma

Overall rate 3/50 (6%) 5/50 (10%) 2 /50 ( 4 %) 0/50 (0%)

Adjusted rate 7.1% 12 .0 % 5. 2% 0 .0 %

Terminal rate 1/23 (4%) 2/26 (8%) 0/18 (0%) 0/13 (0%)

First incidence (days) 646 560 6 14

— Poly

- 3 test P=0.0 75 N P=0.349 P=0. 5 42N P=0.149N



### Testes: Adenoma

Overall rate 34/50 (68%) 38/50 (76%) 32/50 (64%) 29/50 (58%)

Adjusted rate 73.9% 82.7% 75.9% 68.7%

Terminal rate 17/23 (74%) 22/26 (85%) 18/18 (100%) 8/13 (62%)

First incidence (days) 501 528 520 500

Poly

- 3 test P=0.198N P=0.204 P=0.516 P=0.371N



### Thyroid Gland ( C

- Cell ): Adenoma

Overall rate 5/50 (10%) 4/50 (8%) 3/50 (6%) 3/49 (6%)

Adjusted rate 11.9% 9.7% 7.7% 8.5%

Terminal rate 2/23 (9%) 3/26 (12%) 1/18 (6%) 2/13 (15%)

First incidence (days) 660 505 488 710

Poly

- 3 test P=0.356N P=0.510N P=0.397N P=0.451N



### Thyroid Gland ( C

- Cell ): Carcinoma

Overall rate 4/50 (8%) 3/50 (6%) 2/50 (4%) 0/49 (0%)

Adjusted rate 9.7% 7.3% 5.2% 0 .0 %

Terminal rate 4/23 (17%) 2/26 (8%) 1/18 (6%) 0/13 (0%)

First incidence (days) 729 (T) 687 554

— Poly

- 3 test P=0.051N P=0.508N P=0.373N P=0.082N



### Thyroid Gland ( C

- Cell ): Adenoma or Carcinoma

Overall rate 9/50 (18%) 7/50 (14%) 5/50 (10%) 3/49 (6%)

Adjusted rate 21.4% 16.9% 12.6% 8.5%

Terminal rate 6/23 (26%) 5/26 (19%) 2/18 (11%) 2/13 (15%)

First incidence (days) 660 505 488 710

Poly

- 3 test P=0.066N P=0.400N P=0.223N P=0.102N



### Thyroid Gland ( Follicular Cell ): Adenoma or Carcinoma

Overall rate 0/50 (0%) 2/50 (4%) 2/50 (4%) 3/49 (6%)

Adjusted rate 0 .0 % 4.9% 5.2% 8.4%

Terminal rate 0/23 (0%) 2/26 (8%) 1/18 (6%) 2/13 (15%)

First incidence (days)

— 729 (T) 590 687

Poly

- 3 test P=0.075 P=0.233 P=0.219 P=0.093



### All Organs: Malignant Mesothelioma

Overall rate 0/50 (0%) 2/50 (4%) 2/50 (4%) 4/50 (8%)

Adjusted rate 0 .0 % 4.9% 5.2% 10.8%

Terminal rate 0/23 (0%) 2/26 (8%) 0/18 (0%) 1/13 (8%)

First incidence (days)

— 729 (T) 536 394

Poly

- 3 test P=0.031 P=0.233 P=0.220 P=0.046

### 1- Bromopropane, NTP TR 564 93

T ABLE 

### A2

Statistical Analysis of Primary Neoplasms in Male Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

All Organs: Mononuclear Cell Leukemia

Overall rate 29/50 (58%) 22/50 (44%) 16/50 (32%) 24/50 (48%)

Adjusted rate 62.7% 49.8% 40.1% 59.9%

Terminal rate 13/23 (57%) 10/26 (39%) 8/18 (44%) 9/13 (69%)

First incidence (days) 501 540 578 527

Poly

- 3 test P=0.429N P=0.147N P=0.024N P=0.483N



### All Organs: Benign Neoplasms

Overall rate 48/50 (96%) 49/50 (98%) 47/50 (94%) 48/50 (96%)

Adjusted rate 97.6% 99.8% 97.8% 97.9%

Terminal rate 23/23 (100%) 26/26 (100%) 18/18 (100%) 13/13 (100%)

First incidence (days) 429 505 488 394

Poly

- 3 test P=0.611N P=0.444 P=0.794 P=0.760



### All Organs: Malignant Neoplasms

Overall rate 38/50 (76%) 33/50 (66%) 32/50 (64%) 32/50 (64%)

Adjusted rate 79.4% 72.5% 73.5% 74.5%

Terminal rate 18/23 (78%) 19/26 (73%) 15/18 (83%) 11/13 (85%)

First incidence (days) 429 316 488 394

Poly

- 3 test P=0.366N P=0.287N P=0.328N P=0.373N



### All Organs: Benign or Malignant Neoplasms

Overall rate 50/50 (100%) 50/50 (100%) 48/50 (96%) 49/50 (98%)

Adjusted rate 100 .0 % 100 .0 % 98.8% 99.4%

Terminal rate 23/23 (100%) 26/26 (100%) 18/18 (100%) 13/13 (100%)

First incidence (days) 429 316 488 394

Poly

- 3 test P=0.592N

— f P=0.826N P=0.992N

(T) Terminal sacrifice

a Number of neoplasm

- bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal

medulla

, lung, pancreatic islets, pituitary gland, preputial gland, testes, and thyroid gland; for other tissues, denominator is number of animals

necropsied.

b Poly

- 3 estimated neoplasm incidence after adjustment for intercurrent mortality

c Observed incidence at terminal kill

d Beneath the chamber control incidence is the P value associated with the trend test. Ben eath the exposed group incidence are the P values

corresponding to pairwise comparisons between the chamber controls and that expose d group. The Poly

- 3 test accounts for the differential

mortality in animals that do not reach terminal sacrifice. A negati ve trend or a lower incidence in an expos ure group is indicated by N

.

e Not applicable; no neoplasms in animal group

f Value of statistic cannot be computed.

### 94 1- Bromopropane, NTP TR 564

T ABLE 

### A3 a

Historical Incidence of Large Intestine (Colon or Rectum) Neoplasms in Control Male F344/N Rats a



### Study (Study Start) Incidence in Controls

Adenoma Carcinoma Adenoma or Carcinoma

Historical Incidence: Inhalation Studies

1

- Bromopropane (July 2003) 0/50 0/50 0/50

Cumene (June 2001) 0/50 0/50 0/50

Diethylamine (August 2003) 0/50 0/50 0/50

Methyl isobutyl ketone (May 2000) 0/50 0/50 0/50

α

- Methylst y rene (August 2001) 0/50 0/50 0/50

Propargyl alcohol (October 2001) 0/49 0/49 0/49

Tetralin (June 2003) 0/50 0/50 0/50

Total (%) 0/349 0/349 0/349



### Overall Historical Incidence: All Routes

Total (%) 2/1, 398 ( 0.1 %) 1/1, 398 ( 0.1 %) 3/1, 398 (0 .2 %)

Mean 

### ± standard deviation 0.1 % 

### ± 0.5 % 0.1 % 

### ± 0.4 % 0 .2 % ± 0.6 %

Range 0 %

- 2 % 0 %

- 2% 0 %

- 2 %

a Data as of April 29, 2009

### 1- Bromopropane, NTP TR 564 95

T ABLE 

### A3 b

Historical Incidence of Skin Neoplasms in Control Male F344/N Rats a



### Study (Study Start) Incidence in Controls

Keratoacanthoma Basal Cell Adenoma Basal Cell Carcinoma

Historical Incidence: Inhalation Studies

1- Bromopropane (July 2003) 0/50 0 /50 0 /50

Cumene (June 2001) 4/50 1 /50 0 /50

Diethylamine (August 2003) 0/50 0/50 1 /50

Methyl isobutyl ketone (May 2000) 0/50 0/50 0/50

α

- Methylstyrene (August 2001) 1/50 1 /50 0 /50

Propargyl alcohol (October 2001) 4/49 1 /49 0 /49

Tetralin (June 2003) 1/50 1 /50 3 /50

Total (%) 10/349 (2.9%) 4 /349 ( 1.2 %) 4 /349 ( 1.2 %)

Mean 

### ± standard deviation 2.9% 

### ± 3.7% 1.2 .% 

### ± 1.1 % 1.1 .% 

### ± 2.3 %

Range 0%

- 8% 0%

- 2% 0%

- 6 %



### Overall Historical Incidence: All Routes

Total (%) 66/1,398 (4.7%) 15 /1,398 ( 1.1 %) 11 /1,398 ( 0.8 %)

Mean 

### ± standard deviation 4.7% 

### ± 4.2% 1.1 % 

### ± 1.4 % 0.8 % 

### ± 1.5 %

Range 0%

- 16% 0% -4% 0% -6%



### Squamous Cell Carcinoma Keratoacanthoma or

Squamous Cell Carcinoma Keratoacanthoma, Basal

Cell Adenoma, Basal Cell

Carcinoma, or

Squamous Cell Carcinoma

Historical Incidence: Inhalation Studies

1

- Bromopropane (July 2003) 1/50 1 /50 1 /50

Cumene (June 2001) 0 /50 4/50 5 /50

Diethylamine (August 2003) 0/50 0/50 1 /50

Methyl isobutyl ketone (May 2000) 0/50 0/50 0/50

α

- Methylstyrene (August 2001) 0 /50 1/50 2 /50

Propargyl alcohol (October 2001) 0 /49 4/49 5 /49

Tetralin (June 2003) 0 /50 1/50 5 /50

Total (%) 1/349 ( 0.3 %) 1 1 /349 ( 3.2 %) 1 9 /349 ( 5.4 %)

Mean 

### ± standard deviation 0.3 % ± 0.8 % 3.2 % 

### ± 3. 5% 5.5 % 

### ± 4.5 %

Range 0% -2% 0%

- 8% 0%

- 10 %



### Overall Historical Incidence: All Routes

Total (%) 8/1,398 ( 0. 6%) 74 / 1,398 ( 5.3 %) 97 / 1,398 ( 6.9 %)

Mean 

### ± standard deviation 0.6 % ± 0 .9% 5.3 % 

### ± 4.1 % 6.9 % 

### ± 4.9 %

Range 0%

- 2 % 0%

- 16 % 0%

- 20 %

a Data as of April 29, 2009

### 96 1- Bromopropane, NTP TR 564

T ABLE 

### A3 c

Historical Incidence of Malignant Mesothelioma in Control Male F344/N Rats a



### Study (Study Start) Incidence in Controls

Historical Incidence: Inhalation Studies

1- Bromopropane (July 2003) 0/50

Cumene (June 2001) 3/50

Diethylamine (August 2003) 0/50

Methyl isobutyl ketone (May 2000) 1/50

α

- Methylstyrene (August 2001) 0/50

Propargyl alcohol (October 2001) 1/49

Tetralin (June 2003) 0/50

Total (%) 5/349 (1.4%)

Mean 

### ± standard deviation 1.4% 

### ± 2.2%

Range 0%

- 6%



### Overall Historical Incidence: All Routes

Total (%) 35/1,398 (2.5%)

Mean 

### ± standard deviation 2.5% 

### ± 2.3%

Range 0%

- 6%

a Data as of April 29, 2009



### T ABLE 

### A3 d

Historical Incidence of Pancreatic Islet Neoplasms in Control Male F344/N Rats a



### Study (Study Start) Incidence in Controls

Adenoma Carcinoma Adenoma or Carcinoma

Historical Incidence: Inhalation Studies

1

- Bromopropane (July 2003) 0 /50 3 /50 3 /50

Cumene (June 2001) 1 /50 2 /50 3 /50

Diethylamine (August 2003) 6 /50 1 /50 7 /50

Methyl isobutyl ketone (May 2000) 3 /50 4 /50 7 /50

α

- Methylstyrene (August 2001) 3 /50 1/50 4 /50

Propargyl alcohol (October 2001) 4 /49 5 /49 9 /49

Tetralin (June 2003) 3 /50 1 /50 4 /50

Total (%) 20 /349 (5.7%) 17 /349 (4.9%) 37 /349 (10.6%)

Mean 

### ± standard deviation 5.7% 

### ± 3.9% 4.9.% 

### ± 3.3% 10.6% ± 4.8%

Range 0%

- 12% 2%

- 10% 6%

- 18%



### Overall Historical Incidence: All Routes

Total (%) 90 /1,39 4 ( 6.5 %) 29 /1,39 4 ( 2 .1%) 119 /1,39 4 ( 8.5 %)

Mean 

### ± standard deviation 6.5 % 

### ± 3.6 % 2 .1% 

### ± 2.6 % 8.6 % ± 4.0 %

Range 0%

- 14 % 0%

- 10 % 0%

- 18 %

a Data as of April 29, 2009

### 1- Bromopropane, NTP TR 564 97

T ABLE 

### A4

Summary of the Incidence of Nonneoplastic Lesions in Male Rats

in the 2

- Year Inhalation Study of 1- Bromopropane a



### Chamber Control 125 ppm 250 ppm 500 ppm

Disposition Summary

Animals initially in study 50 50 50 50

Early deaths

Moribund 24 20 28 35

Natural deaths 3 4 4 2

Survivors

Terminal sacrifice 23 26 18 13

Animals examined microscopically 50 50 50 50



### Alimentary System

Esophagus (50) (50) (50) (50)

Intestine l arge, c ecum (47) (47) (47) (48)

Edema 1 (2%)

Inflammation, c hronic a ctive 1 (2%)

Necrosis 1 (2%)

Artery, i nflammation 1 (2%)

Intestine l arge, c olon (48) (48) (48) (48)

Artery, i nflammation 1 (2%)

Intestine l arge, r ectum (48) (48) (48) (49)

Necrosis 1 (2%) 1 (2%)

Intestine s mall, d uodenum (48) (48) (48) (48)

Fibrosis 1 (2%)

Intestine s mall, i leum (47) (47) (46) (48)

Inflammation, c hronic a ctive 1 (2%)

Necrosis 1 (2%) 1 (2%)

Intestine s mall, j ejunum (47) (47) (46) (48)

Liver (50) (50) (50) (50)

Angiectasis 2 (4%) 1 (2%)

Basophilic f ocus 9 (18%) 9 (18%) 8 (16%) 8 (16%)

Clear c ell f ocus 16 (32%) 25 (50%) 17 (34%) 15 (30%)

Degeneration, c ystic 2 (4%) 2 (4%) 1 (2%) 4 (8%)

Eosinophilic f ocus 1 (2%) 1 (2%) 1 (2%)

Hemorrhage 1 (2%)

Hepatodiaphragmatic n odule 4 (8%) 1 (2%) 2 (4%)

Mixed c ell f ocus 2 (4%) 1 (2%) 3 (6%)

Necrosis 2 (4%) 1 (2%) 1 (2%) 1 (2%)

Thrombosis 1 (2%) 1 (2%)

Vacuolization c ytoplasmic 31 (62%) 38 (76%) 32 (64%) 37 (74%)

Serosa, f ibrosis 1 (2%)

Mesentery (13) (13) (8) (19)

Necrosis 13 (100%) 13 (100%) 7 (88%) 17 (89%)

Oral m ucosa (3) (0) (2) (4)

Foreign b ody 1 (33%) 1 (50%) 2 ( 50%)

Hyperplasia, s quamous 1 (33%) 1 (50%)

Inflammation, c hronic a ctive 1 (33%) 2 (50%)

Pancreas (50) (50) (50) (50)

Basophilic f ocus 1 (2%) 1 (2%)

Cyst 1 (2%)

Fibrosis 1 (2%)

Necrosis 1 (2%)

Acinus, a trophy 23 (46%) 13 (26%) 20 (40%) 23 (46%)

Artery, i nflammation 1 (2%) 1 (2%)

Salivary g lands (50) (50) (50) (50)

Hyperplasia 1 (2%)

Inflammation, c hronic a ctive 1 (2%) 3 (6%) 2 (4%)

a Number of animals examined microscopically at the site and the number of animals with lesion

### 98 1- Bromopropane, NTP TR 564

T ABLE 

### A4

Summary of the Incidence of Nonneoplastic Lesions in Male Rats

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Alimentary System (continued)

Stomach, f orestomach (50) (50) (50) (50)

Erosion 1 (2%)

Hyperplasia, s quamous 1 (2%) 1 (2%) 1 (2%) 1 (2%)

Ulcer 7 (14%) 3 (6%) 5 (10%) 4 (8%)

Stomach, g landular (50) (50) (50) (50)

Erosion 1 (2%) 1 (2%)

Ulcer 1 (2%) 2 (4%) 1 (2%)

Artery, i nflammation 1 (2%)

Epithelium, h yperplasia 1 (2%)

Tongue (0) (0) (1) (1)

Hyperplasia, s quamous 1 (100%) 1 (100%)



### Cardiovascular System

Heart (50) (50) (50) (50)

Cardiomyopathy 26 ( 52%) 23 (46%) 23 (46%) 15 (30%)

Atrium, t hrombosis 4 (8%) 2 (4%)

Ventricle, t hrombosis 1 (2%)



### Endocrine System

Adrenal c ortex (50) (50) (50) (50)

Hyperplasia 27 (54%) 21 (42%) 22 (44%) 20 (40%)

Vacuolization c ytoplasmic 9 (18%) 16 (32%) 9 (18%) 12 (24%)

Adrenal m edulla (50) (50) (50) (50)

Hemorrhage 1 (2%)

Hyperplasia 23 (46%) 18 (36%) 23 (46%) 19 (38%)

Islets, p ancreatic (50) (50) (50) (50)

Hyperplasia 10 (20%) 7 (14%) 6 (12%) 9 (18%)

Parathyroid g land (46) (48) (47) (46)

Hyperplasia 2 (4%)

Pituitary g land (50) (50) (50) (49)

Cyst 1 (2%) 2 (4%) 1 (2%)

Hemorrhage 4 (8%) 2 (4%) 5 (10%)

Pars d istalis, h yperplasia 8 (16%) 5 (10%) 11 (22%) 7 (14%)

Thyroid g land (50) (50) (50) (49)

Ultimobranchial c yst 1 (2%)

C- cell, h yperplasia 32 (64%) 31 (62%) 29 (58%) 34 (69%)

Follicular c ell, h yperplasia 1 ( 2%)



### General Body System

Peritoneum (0) (0) (0) (2)

Inflammation, s uppurative, c hronic 1 (50%)



### Genital System

Coagulating g land (3) (0) (1) (2)

Hyperplasia 1 (33%)

Inflammation, s uppurative 2 (67%)

Epididymis (50) (50) (50) (50)

Penis (0) (3) (1) (1)

Concretion 1 (100%)

Hyperplasia, s quamous 1 (33%)

### 1- Bromopropane, NTP TR 564 99

T ABLE 

### A4

Summary of the Incidence of Nonneoplastic Lesions in Male Rats

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Genital System (continued)

Preputial g land (50) (50) (49) (50)

Ectasia 1 (2%) 1 (2%)

Hyperplasia 3 (6%) 1 (2%)

Inflammation, s uppurative, c hronic 1 (2%)

Inflammation, c hronic a ctive 30 (60%) 33 (66%) 31 (63%) 37 (74%)

Prostate (50) (50) (50) (50)

Hyperplasia 12 (24%) 13 (26%) 10 (20%) 17 (34%)

Inflammation, s uppurative, c hronic 1 (2%)

Inflammation, s uppurative 38 (76%) 42 (84%) 40 (80%) 36 (72%)

Seminal v esicle (50) (49) (49) (50)

Congestion 1 (2%)

Dilatation 1 (2%)

Hyperplasia 1 (2%)

Inflammation, s uppurative 2 (4 %) 1 (2%) 2 (4%)

Testes (50) (50) (50) (50)

Mineralization 1 (2%) 1 (2%)

Germinal e pithelium, a trophy 12 (24%) 8 (16%) 7 (14%) 6 (12%)

Interstitial c ell, h yperplasia 1 (2%) 2 (4%) 1 (2%)



### Hematopoietic System

Bone m arrow (50) (50) (50) (50)

Thrombosis 1 (2%)

Lymph n ode (5) (5) (5) (4)

Deep c ervical, a ngiectasis 1 (20%)

Pancreatic, a ngiectasis 2 (40%)

Pancreatic, h emorrhage 1 (25%)

Lymph n ode, b ronchial (3) (6) (5) (8)

Angiectasis 1 (33%) 2 (33%) 2 (40%)

Fibrosis 1 (17%)

Hemorrhage 1 (33%) 1 (17%) 1 (20%) 1 (13%)

Hyperplasia, l ymphoid 4 (50%)

Lymph n ode, m andibular (3) (1) (0) (6)

Angiectasis 1 (17%)

Lymph n ode, m ediastinal (30) (25) (23) (32)

Angiectasis 1 (4%) 3 (9%)

Hyperplasia, l ymphoid 1 (4%) 1 (4%) 2 (6%)

Lymph n ode, m esenteric (50) (50) (50) (50)

Angiectasis 1 (2%) 1 (2%) 2 (4%)

Hyperplasia, l ymphoid 1 (2%)

Infiltration c ellular, h istiocyte 1 (2%)

Artery, n ecrosis 1 (2%)

Spleen (50) (50) (49) (50)

Accessory s pleen 1 (2%) 1 (2%)

Fibrosis 1 (2%) 2 (4%) 1 (2%)

Hematopoietic c ell p roliferation 10 (20%) 17 (34%) 13 (27%) 6 (12%)

Hemorrhage, c hronic 4 (8%) 1 (2%) 3 (6%)

Hyperplasia, l ymphoid 1 (2%)

Infarct, c hronic 1 (2%) 2 (4%) 3 (6%)

Infiltration c ellular, m ononuclear c ell 1 (2%)

Thymus (43) (41) (47) (46)

Cyst 1 (2%)

### 100 1- Bromopropane, NTP TR 564

T ABLE 

### A4

Summary of the Incidence of Nonneoplastic Lesions in Male Rats

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Integumentary System

Mammary g land (40) (36) (39) (36)

Galactocele 1 (3%) 1 (3%)

Hyperplasia 1 (3%)

Epithelium, h yperplasia 1 (3%) 1 (3%) 1 (3%)

Skin (50) (50) (50) (50)

Cyst e pithelial i nclusion 1 (2%) 1 (2%)

Foreign b ody 1 (2%)

Hyperkeratosis 3 (6%) 2 (4%)

Hyperplasia, s quamous 1 (2%)

Inflammation, s uppurative, c hronic 1 (2%) 2 (4%) 10 (20%)

Inflammation, c hronic a ctive 1 (2%) 3 (6%) 3 (6%) 4 (8%)

Thrombosis 1 (2%)



### Musculoskeletal System

Bone (50) (50) (50) (50)

Fibrosis 1 (2%)

Hyperostosis 1 (2%)

Inflammation, s uppurative, c hronic 2 (4%)

Skeletal m uscle (1) (1) (3) (3)

Inflammation, s uppurative, c hronic 1 (33%)



### Nervous System

Brain (50) (50) (50) (50)

Hemorrhage 1 (2%) 1 (2%) 2 (4%) 2 (4%)

Spinal c ord (1) (1) (2) (1)



### Respiratory System

Larynx (50) (50) (50) (50)

Foreign b ody 6 (12%) 1 (2%) 5 (10%) 1 (2%)

Inflammation, s uppurative, c hronic 1 (2%)

Inflammation, c hronic a ctive 21 (42%) 28 (56%) 31 (62%) 26 (52%)

Metaplasia, s quamous 4 (8%) 6 (12%) 8 (16%) 5 (10%)

Respiratory e pithelium, h yperplasia 1 (2%) 1 (2%) 1 (2%)

Lung (50) (50) (50) (50)

Foreign b ody 1 (2%)

Hemorrhage 2 (4%) 5 (10%) 1 (2%) 1 (2%)

Inflammation, s uppurative, c hronic 1 (2%) 3 (6%)

Inflammation, c hronic a ctive 5 (10%) 3 (6%)

Metaplasia, o sseous 1 (2%)

Alveolar e pithelium, h yperplasia 15 (30%) 11 (22%) 13 (26%) 11 (22%)

Alveolar e pithelium, m etaplasia, s quamous 1 (2%)

Alveolus, i nfiltration c ellular, h istiocyte 16 ( 32%) 14 ( 28%) 13 (26%) 11 (22%)

Artery, i nflammation 1 (2%)

### 1- Bromopropane, NTP TR 564 101

T ABLE 

### A4

Summary of the Incidence of Nonneoplastic Lesions in Male Rats

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Respiratory System (continued)

Nose (50) (48) (48) (50)

Degeneration, h yaline 1 (2%)

Foreign b ody 11 (22%) 10 (21%) 15 (31%) 11 (22%)

Inflammation, s uppurative, c hronic 1 (2%) 2 (4%) 7 (14%)

Inflammation, c hronic a ctive 29 (58%) 33 (69%) 34 (71%) 35 (70%)

Epithelium, a ccumulation, h yaline d roplet 44 (88%) 39 (81%) 36 (75%) 44 (88%)

Glands, h yperplasia 5 (10%) 14 (29%) 14 (29%) 15 ( 30%)

Olfactory e pithelium, a trophy 1 (2%)

Olfactory e pithelium, m etaplasia,

r espiratory 7 (14%) 10 (21%) 12 (25%) 12 (24%)

Respiratory e pithelium, h yperplasia 14 (28%) 15 (31%) 20 (42%) 17 (34%)

Respiratory e pithelium, m etaplasia,

squamous 1 (2%) 1 (2%) 2 (4%)

Trachea (50) (50) (50) (50)

Inflammation, c hronic a ctive 1 (2%) 1 (2%) 1 (2%) 4 (8%)

Epithelium, h yperplasia 1 (2%) 1 (2%)



### Special Senses System

Eye (49) (49) (50) (50)

Inflammation, c hronic a ctive 1 (2%) 2 (4%)

Lens, c ataract 2 (4%) 3 (6%) 2 (4%)

Retina, d egeneration 1 (2%)

Harderian g land (50) (50) (50) (50)

Inflammation, s uppurative, c hronic 2 (4%)

Lacrimal g land (1) (0) (0) (0)

Inflammation, c hronic a ctive 1 (100%)

Zymbal ’ s g land (0) (1) (2) (2)



### Urinary System

Kidney (50) (50) (50) (49)

Cyst 1 (2%) 1 (2%)

Inflammation, s uppurative 1 (2%)

Nephropathy, c hronic 44 (88%) 45 (90%) 39 (78%) 44 (90%)

Cortex, i nfarct 1 (2%)

Cortex, r enal t ubule, c asts g ranular 2 (4%)

Pelvis, i nflammation, c hronic a ctive 2 (4%)

Urinary b ladder (50) (50) (50) (50)

Hemorrhage 1 (2%)

Inflammation, c hronic a ctive 1 (2%) 1 (2%)

Transitional e pithelium, h yperplasia 1 (2%)

### 102 1- Bromopropane, NTP TR 564

### 103



# APPENDIX B

SUMMARY OF LESIONS IN FEMALE RA TS

IN THE 2

- YEAR INHALATION STUDY

OF 1- BROMOPROPANE

T ABLE 

### B1 Summary of the Incidence of Neoplasms in Fem ale Rats

in the 2

- Year Inhalation Study of 1- Bromopropane

....................................................................... 104

T ABLE 

### B2 Statistical Analysis of Primary Neoplasms in Fem ale Rats

in the 2

- Year Inhalation Study of 1- Bromopropane

....................................................................... 108

T ABLE 

### B3a Historical Incidence of Large Intestine (Colon or Rectum) Neoplasms

i n Control Female F344/N Rats

......................................................................................................... 112

T ABLE 

### B3b Historical Incidence of Skin Neoplasms in Control Female F344/N Rats

..................................... 113

T ABLE 

### B4 Summary of the Incidence of Nonneoplastic Lesions in Fem ale Rats

in the 2

- Year Inhalation Study of 1

- Bromopropane

...................................................................... 114

### 104 1- Bromopropane, NTP TR 564

T ABLE 

### B1

Summary of the Incidence of Ne oplas ms in Fem ale Rats in the 2

- Year Inhalation Study of 1- Bromopropane a



### Chamber Control 125 ppm 250 ppm 500 ppm

Disposition Summary

Animals initially in study 50 50 50 50

Early deaths

Accidental deaths 2

Moribund 13 17 17 23

Natural deaths 3 3 1

Survivors

Terminal sacrifice 34 33 30 24

Animals examined microscopically 50 50 50 50



### Alimentary System

Intestine l arge, c olon (47) (50) (48) (49)

Adenoma 1 (2%) 1 (2%) 1 (2%)

Carcinoid t umor m alignant 1 (2%)

Intestine l arge, r ectum (47) (50) (48) (49)

Adenoma 1 (2%) 4 (8%)

Intestine s mall, i leum (46) (50) (47) (48)

Carcinoma 1 (2%)

Intestine s mall, j ejunum (47) (50) (47) (48)

Liver (50) (50) (50) (50)

Hepatocellular c arcinoma 1 (2%)

Mesentery (13) (26) (17) (23)

Sarcoma 1 (6%)

Sarcoma, m etastatic, s tomach, forestomach 1 (4%)

Oral m ucosa (2) (0) (1) (1)

Pancreas (50) (50) (50) (50)

Sarcoma, m etastatic, m esentery 1 (2%)

Salivary g lands (50) (50) (50) (50)

Stomach, f orestomach (50) (50) (50) (50)

Sarcoma 1 (2%)

Stomach, g landular (50) (50) (50) (50)

Sarcoma 1 (2%)

Tongue (0) (1) (0) (1)

Tooth (0) (0) (1) (0)



### Cardiovascular System

Heart (50) (50) (50) (50)

Alveolar/ b ronchiolar c arcinoma, metastatic,

lung 1 (2%)



### Endocrine System

Adrenal c ortex (50) (50) (50) (50)

Adenoma 3 (6%) 3 (6%)

Adrenal m edulla (50) (50) (50) (50)

Pheochromocytoma b enign 2 (4%) 4 (8%) 1 (2%)

Pheochromocytoma c omplex 1 (2%)

Islets, p ancreatic (50) (50) (50) (50)

Adenoma 2 (4%) 1 (2%)

Carcinoma 1 (2%) 1 (2%) 1 (2%)

Pituitary g land (50) (50) (50) (50)

Pars d istalis, a denoma 35 (70%) 33 (66%) 28 (56%) 30 (60%)

Pars d istalis, c arcinoma 1 (2%) 1 (2%)

### 1- Bromopropane, NTP TR 564 105

T ABLE 

### B1

Summary of the Incidence of Ne oplas ms in Fem ale Rats in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Endocrine System (continued)

Thyroid g land (50) (50) (50) (50)

C

- cell, a denoma 3 (6%) 3 (6%) 3 (6%) 4 (8%)

C

- cell, c arcinoma 1 (2%) 2 (4%) 2 (4%) 2 (4%)

Follicular c ell, a denoma 1 (2%) 1 (2%)

Follicular c ell, c arcinoma 1 (2%)



### General Body System

None



### Genital System

Clitoral g land (49) (50) (49) (49)

Adenoma 3 (6%) 9 (18%) 3 (6%) 3 (6%)

Carcinoma 1 (2%) 3 (6%) 2 (4%) 1 (2%)

Ovary (50) (50) (50) (50)

Cystadenoma 1 (2%)

Granulosa c ell t umor m alignant 1 (2%)

Granulosa

- t heca t umor m alignant 2 (4%)

Uterus (50) (50) (50) (50)

Adenoma 1 (2%)

Polyp s tromal 8 (16%) 8 (16%) 8 (16%) 7 (14%)

Sarcoma s tromal 2 (4%) 1 (2%) 1 (2%)

Vagina (3) (0) (0) (0)

Sarcoma 1 (33%)



### Hematopoietic System

Bone m arrow (50) (50) (50) (50)

Lymph n ode (1) (3) (2) (2)

Lymph n ode, b ronchial (1) (4) (5) (7)

Squamous c ell c arcinoma, m etastatic, lung 1 (25%)

Lymph n ode, m ediastinal (34) (30) (31) (28)

Carcinoma, m etastatic, t hyroid g land 1 (3%)

Lymph n ode, m esenteric (49) (49) (50) (50)

Spleen (50) (50) (50) (50)

Thymus (46) (45) (45) (43)

Thymoma b enign 1 (2%)



### Integumentary System

Mammary g land (50) (50) (50) (50)

Carcinoma 4 (8%) 3 (6%) 3 (6%) 1 (2%)

Fibroadenoma 19 (38%) 14 (28%) 15 (30%) 15 (30%)

Fibroadenoma, m ultiple 6 (12%) 6 (12%) 6 (12%) 1 (2%)

Skin (50) (50) (50) (50)

Basal c ell a denoma 1 (2%) 1 (2%)

Basal c ell c arcinoma 1 (2%)

Keratoacanthoma 1 (2%) 1 (2%) 1 (2%)

Squamous c ell p apilloma 1 (2%)

Pinna, n eural c rest t umor 1 (2%) 1 (2%)

Sebaceous g land, a denoma 1 (2%)

Subcutaneous t issue, f ibroma 2 (4%)

Subcutaneous t issue, l ipoma 1 (2%)

### 106 1- Bromopropane, NTP TR 564

T ABLE 

### B1

Summary of the Incidence of Ne oplas ms in Fem ale Rats in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Musculoskeletal System

Bone (50) (50) (50) (50)

Skeletal m uscle (0) (0) (1) (4)

Alveolar/ b ronchiolar c arcinoma, metastatic,

lung 1 (100%)



### Nervous System

Brain (50) (50) (50) (50)

Carcinoma, m etastatic, p ituitary g land 1 (2%) 1 (2%)

Glioma m alignant 1 (2%)

Spinal c ord (0) (1) (0) (3)



### Respiratory System

Larynx (50) (50) (50) (50)

Lung (50) (50) (50) (50)

Alveolar/ b ronchiolar a denoma 1 (2%) 4 (8%) 2 (4%)

Alveolar/ b ronchiolar a denoma, m ultiple 1 (2%)

Alveolar/ b ronchiolar c arcinoma 1 (2%) 1 (2%) 2 (4%)

Squamous c ell c arcinoma 1 (2%)

Nose (50) (50) (49) (50)

Fibroma 1 (2%)

Pleura (0) (0) (1) (0)

Alveolar/ b ronchiolar c arcinoma, metastatic,

lung 1 (100%)

Trachea (50) (50) (50) (50)



### Special Senses System

Ear (0) (0) (1) (1)

Eye (50) (50) (50) (50)

Harderian g land (50) (50) (50) (50)

Zymbal ’ s g land (1) (1) (2) (0)

Carcinoma 1 (100%) 1 (100%) 2 (100%)



### Urinary System

Kidney (49) (50) (50) (50)

Sarcoma, m etastatic, s tomach, forestomach 1 (2%)

Renal t ubule, c arcinoma 1 (2%)



### Systemic Neoplasms

Multiple o rgans b (50) (50) (50) (50)

Histiocytic s arcoma 1 (2%)

Leukemia m ononuclear 16 (32%) 13 (26%) 17 (34%) 15 (30%)

Lymphoma m alignant 1 (2%)

### 1- Bromopropane, NTP TR 564 107

T ABLE 

### B1

Summary of the Incidence of Ne oplas ms in Fem ale Rats in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Neoplasm Summary

Total a nimals with p rimary n eoplasms c 49 48 46 43

Total p rimary n eoplasms 119 120 106 96

Total a nimals with b enign n eoplasms 45 46 36 41

Total b enign n eoplasms 84 90 72 72

Total a nimals with m alignant neoplasms 26 23 28 20

Total m alignant n eoplasms 34 29 34 24

Total a nimals with m etastatic neoplasms 2 4 1

Total m etastatic n eoplasms 2 6 2

Total a nimals with uncertain n eoplasms

-

b enign or m alignant 1 1

Total u ncertain n eoplasms 1 1

a Number of animals examined microscopically at the site and the number of animals with neoplasm

b Number of animals with any tissue examined microscopically

c Primary neoplasms: all neoplasms except metastatic neoplasms

### 108 1- Bromopropane, NTP TR 564

T ABLE 

### B2

Statistical Analysis of Primary Neoplasms in Fem ale Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Adrenal Cortex : Adenoma

Overall rate a 3/50 (6%) 3/50 (6%) 0/50 (0%) 0/50 (0%)

Adjusted rate b 6.6% 6.9% 0 .0 % 0 .0 %

Terminal rate c 2/34 (6%) 3/33 (9%) 0/30 (0%) 0/24 (0%)

First incidence (days) 701 730 (T)

— e

— Poly

- 3 test d P=0.042N P=0.645 P=0.131N P=0.156N



### Adrenal Medulla : Benign Pheochromocytoma

Overall rate 2/50 (4%) 4/50 (8%) 1/50 (2%) 0/50 (0%)

Adjusted rate 4.4% 9.1% 2.3% 0 .0 %

Terminal rate 2/34 (6%) 3/33 (9%) 0/30 (0%) 0/24 (0%)

First incidence (days) 730 (T) 687 607

— Poly

- 3 test P=0.118N P=0.323 P=0.519N P=0.281N



### Adrenal Medulla: Benign or Complex Pheochromocytoma

Overall rate 3/50 (6%) 4/50 (8%) 1/50 (2%) 0/50 (0%)

Adjusted rate 6.6% 9.1% 2.3% 0 .0 %

Terminal rate 2/34 (6%) 3/33 (9%) 0/30 (0%) 0/24 (0%)

First incidence (days) 677 687 607

— Poly

- 3 test P=0.062N P=0.481 P=0.327N P=0.156N



### Clitoral Gland: Adenoma

Overall rate 3/49 (6%) 9/50 (18%) 3/49 (6%) 3/49 (6%)

Adjusted rate 6.7% 20.5% 7.2% 8 .0 %

Terminal rate 2/33 (6%) 8/33 (24%) 3/30 (10%) 1/24 (4%)

First incidence (days) 677 719 730 (T) 627

Poly

- 3 test P=0.390N P=0.054 P=0.631 P=0.583



### Clitoral Gland: Carcinoma

Overall rate 1/49 (2%) 3/50 (6%) 2/49 (4%) 1/49 (2%)

Adjusted rate 2.3% 6.8% 4.8% 2.7%

Terminal rate 1/33 (3%) 2/33 (6%) 2/30 (7%) 0/24 (0%)

First incidence (days) 730 (T) 632 730 (T) 724

Poly

- 3 test P=0.556N P=0.303 P=0.478 P=0.720



### Clitoral Gland: Adenoma or Carcinom a

Overall rate 4/49 (8%) 12/50 (24%) 4/49 (8%) 4/49 (8%)

Adjusted rate 9 .0 % 27.2% 9.6% 10.6%

Terminal rate 3/33 (9%) 10/33 (30%) 4/30 (13%) 1/24 (4%)

First incidence (days) 677 632 730 (T) 627

Poly

- 3 test P=0.353N P=0.023 P=0.606 P=0.549



### Large Intestine (Colon and Rectum): Adenoma

Overall rate 0/50 (0%) 1/50 (2%) 2/50 (4%) 5/50 (10%)

Adjusted rate 0 .0 % 2.3% 4.7% 13.3%

Terminal rate 0/34 (0%) 1/33 (3%) 1/30 (3%) 4/24 (17%)

First incidence (days)

— 730 (T) 607 719

Poly -3 test P=0.004 P=0.493 P=0.225 P=0.018



### Lung: Alveolar/ b ronchiolar Adenoma

Overall rate 1/50 (2%) 4/50 (8%) 1/50 (2%) 2/50 (4%)

Adjusted rate 2.2% 9.1% 2.4% 5.3%

Terminal rate 1/34 (3%) 3/33 (9%) 1/30 (3%) 0/24 (0%)

First incidence (days) 730 (T) 635 730 (T) 627

Poly

- 3 test P=0.509 P=0.170 P=0.746 P=0.440

### 1- Bromopropane, NTP TR 564 109

T ABLE 

### B2

Statistical Analysis of Primary Neoplasms in Fem ale Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Lung: Alveolar/ b ronchiolar Adenoma or Carcinoma

Overall rate 2/50 (4%) 5/50 (10%) 3/50 (6%) 2/50 (4%)

Adjusted rate 4.4% 11.3% 6.9% 5.3%

Terminal rate 1/34 (3%) 3/33 (9%) 1/30 (3%) 0/24 (0%)

First incidence (days) 694 635 417 627

Poly

- 3 test P=0.521N P=0.205 P=0.482 P=0.628



### Mammary Gland: Fibroadenoma

Overall rate 25/50 (50%) 20/50 (40%) 21/50 (42%) 16/50 (32%)

Adjusted rate 53.8% 42.8% 48.4% 41.3%

Terminal rate 18/34 (53%) 12/33 (36%) 16/30 (53%) 11/24 (46%)

First incidence (days) 638 528 642 501

Poly -3 test P=0.200N P=0.194N P=0.383N P=0.172N



### Mammary Gland: Carcinoma

Overall rate 4/50 (8%) 3/50 (6%) 3/50 (6%) 1/50 (2%)

Adjusted rate 8.7% 6.8% 7 .0 % 2.7%

Terminal rate 0/34 (0%) 2/33 (6%) 2/30 (7%) 1/24 (4%)

First incidence (days) 612 575 684 730 (T)

Poly

- 3 test P=0.196N P=0.524N P=0.544N P=0.248N



### Mammary Gland: Fibroadenoma or Carcinoma

Overall rate 27/50 (54%) 20/50 (40%) 23/50 (46%) 16/50 (32%)

Adjusted rate 57.1% 42.8% 53.1% 41.3%

Terminal rate 18/34 (53%) 12/33 (36%) 18/30 (60%) 11/24 (46%)

First incidence (days) 612 528 642 501

Poly

- 3 test P=0.149N P=0.115N P=0.428N P=0.102N



### Pancreatic Islets: Adenoma or Carcinoma

Overall rate 1/50 (2%) 3/50 (6%) 2/50 (4%) 0/50 (0%)

Adjusted rate 2.2% 6.9% 4.7% 0 .0 %

Terminal rate 1/34 (3%) 3/33 (9%) 2/30 (7%) 0/24 (0%)

First incidence (days) 730 (T) 730 (T) 730 (T)

— Poly

- 3 test P=0.315N P=0.293 P=0.477 P=0.537N



### Pituitary Gland (Pars Distalis): Adenoma

Overall rate 35/50 (70%) 33/50 (66%) 28/50 (56%) 30/50 (60%)

Adjusted rate 73.9% 68.2% 62 .0 % 71.3%

Terminal rate 25/34 (74%) 21/33 (64%) 20/30 (67%) 17/24 (71%)

First incidence (days) 633 433 534 450

Poly

- 3 test P=0.418N P=0.347N P=0.149N P=0.485N



### Pituitary Gland (Pars Distalis): Adenoma or Carcinoma

Overall rate 35/50 (70%) 34/50 (68%) 29/50 (58%) 30/50 (60%)

Adjusted rate 73.9% 70.3% 64.1% 71.3%

Terminal rate 25/34 (74%) 22/33 (67%) 20/30 (67%) 17/24 (71%)

First incidence (days) 633 433 534 450

Poly

- 3 test P=0.404N P=0.433N P=0.206N P=0.485N



### Skin: Squamous Cell Papilloma, Keratoacanthoma

, Basal Cell Adenoma, or Basal Cell Carcinoma

Overall rate 1/50 (2%) 1/50 (2%) 1/50 (2%) 4/50 (8%)

Adjusted rate 2.2% 2.3% 2.4% 10.6%

Terminal rate 1/34 (3%) 1/33 (3%) 1/30 (3%) 4/24 (17%)

First incidence (days) 730 (T) 730 (T) 730 (T) 730 (T)

Poly

- 3 test P=0.050 P=0.753 P=0.746 P=0.128

### 110 1- Bromopropane, NTP TR 564

T ABLE 

### B2

Statistical Analysis of Primary Neoplasms in Fem ale Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Thyroid Gland (C

- Cell): Adenoma

Overall rate 3/50 (6%) 3/50 (6%) 3/50 (6%) 4/50 (8%)

Adjusted rate 6.6% 6.8% 7 .0 % 10.6%

Terminal rate 3/34 (9%) 2/33 (6%) 2/30 (7%) 4/24 (17%)

First incidence (days) 730 (T) 659 694 730 (T)

Poly

- 3 test P=0.310 P=0.649 P=0.634 P=0.400



### Thyroid Gland (C

- Cell): Adenoma or Carcinoma

Overall rate 4/50 (8%) 5/50 (10%) 5/50 (10%) 6/50 (12%)

Adjusted rate 8.8% 11.4% 11.7% 15.8%

Terminal rate 4/34 (12%) 4/33 (12%) 4/30 (13%) 5/24 (21%)

First incidence (days) 730 (T) 659 694 626

Poly

- 3 test P=0.212 P=0.482 P=0.462 P=0.264



### Uterus: Stromal Polyp

Overall rate 8/50 (16%) 8/50 (16%) 8/50 (16%) 7/50 (14%)

Adjusted rate 17.3% 18 .0 % 18.5% 17.9%

Terminal rate 6/34 (18%) 6/33 (18%) 5/30 (17%) 3/24 (13%)

First incidence (days) 438 577 607 590

Poly

- 3 test P=0.519 P=0.573 P=0.551 P=0.584



### Uterus: Stromal Polyp or Stromal Sarcoma

Overall rate 10/50 (20%) 8/50 (16%) 8/50 (16%) 8/50 (16%)

Adjusted rate 21.3% 18 .0 % 18.5% 20.3%

Terminal rate 7/34 (21%) 6/33 (18%) 5/30 (17%) 3/24 (13%)

First incidence (days) 438 577 607 590

Poly

- 3 test P=0.516N P=0.449N P=0.472N P=0.558N



### All Organs: Mononuclear Cell Leukemia

Overall rate 16/50 (32%) 13/50 (26%) 17/50 (34%) 15/50 (30%)

Adjusted rate 34.3% 29.2% 37.6% 38.7%

Terminal rate 9/34 (27%) 10/33 (30%) 8/30 (27%) 10/24 (42%)

First incidence (days) 633 590 541 611

Poly

- 3 test P=0.294 P=0.385N P=0.453 P=0.423



### All Organs: Benign Neoplasms

Overall rate 45/50 (90%) 46/50 (92%) 36/50 (72%) 41/50 (82%)

Adjusted rate 93.3% 92.2% 79.3% 92.9%

Terminal rate 33/34 (97%) 30/33 (91%) 26/30 (87%) 23/24 (96%)

First incidence (days) 438 433 534 450

Poly

- 3 test P=0.399N P=0.577N P=0.028N P=0.654N



### All Organs: Malignant Neoplasms

Overall rate 26/50 (52%) 23/50 (46%) 28/50 (56%) 20/50 (40%)

Adjusted rate 53.3% 50 .0 % 58.9% 50.4%

Terminal rate 13/34 (38%) 15/33 (46%) 15/30 (50%) 12/24 (50%)

First incidence (days) 493 575 417 578

Poly

- 3 test P=0.519N P=0.452N P=0.363 P=0.475N

### 1- Bromopropane, NTP TR 564 111

T ABLE 

### B2

Statistical Analysis of Primary Neoplasms in Fem ale Rats in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

All Organs: Benign or Malignant Neoplasms

Overall rate 49/50 (98%) 48/50 (96%) 46/50 (92%) 43/50 (86%)

Adjusted rate 98 .0 % 96 .0 % 92.7% 96.6%

Terminal rate 33/34 (97%) 31/33 (94%) 27/30 (90%) 24/24 (100%)

First incidence (days) 438 433 417 450

Poly

- 3 test P=0.415N P=0.500N P=0.211N P=0.613N

(T) Terminal sacrifice

a Number of neoplasm

- bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal

gland , clitoral gland, lung, pancreatic islets, pituitary gland, and thyroid gland; for other tissues, denominator is numb er of animals necropsied.

b Poly

- 3 estimated neoplasm incidence after adjustment for intercurrent mortality

c Observed incidence at terminal kill

d Beneath the chamber control incidence is the P value associated with the trend test. Beneath the exposed gr oup incidence are the P values

corresponding to pairwise comparisons between the chamber controls and that expos ed group. The Poly

- 3 test accounts for the differential

mortality in animals that do not reach terminal sacrifice. A negative trend or a lower incidence in an expos ure group is indicated by N

.

e Not applicable; no neoplasms in animal group

### 112 1- Bromopropane, NTP TR 564

T ABLE 

### B3 a

Historical Incidence of Large Intestine (Colon or Rectum) Neoplasms in Control Female F344/N Rats a



### Study (Study Start) Incidence in Controls

Adenoma Carcinoma Adenoma or Carcinoma

Historical Incidence: Inhalation Studies

1

- Bromopropane (July 2003) 0/50 0/50 0/50

Cumene (June 2001) 0/50 0/50 0/50

Diethylamine (August 2003) 0/50 0/50 0/50

Methyl isobutyl ketone (May 2000) 0/50 0/50 0/50

α

- Methylst y rene (August 2001) 0/50 0/50 0/50

Propargyl alcohol (October 2001) 0/50 0/50 0/50

Tetralin (June 2003) 0/50 0/50 0/50

Total (%) 0/3 50 0/3 50 0/3 50



### Overall Historical Incidence: All Routes

Total (%) 3 /1,3 50 (0. 2 %) 1/1,3 50 (0.1%) 4 /1,3 50 (0. 3 %)

Mean 

### ± standard deviation 0. 2% 

### ± 0. 6% 0.1% 

### ± 0.4% 0. 3 % ± 0. 7 %

Range 0%

- 2% 0%

- 2% 0%

- 2%

a Data as of April 29, 2009

### 1- Bromopropane, NTP TR 564 113

T ABLE 

### B3b

Historical Incidence of Skin Neoplasms in Control Fem ale F344/N Rats a



### Study (Study Start) Incidence in Controls

Squamous Cell Papilloma Keratoacanthoma Basal Cell Adenoma

Historical Incidence: Inhalation Studies

1- Bromopropane (July 2003) 0/50 1 /50 0 /50

Cumene (June 2001) 0 /50 0 /50 0 /50

Diethylamine (August 2003) 0/50 1 /50 0/50

Methyl isobutyl ketone (May 2000) 0/50 0/50 0/50

α

- Methylstyrene (August 2001) 0 /50 0 /50 0 /50

Propargyl alcohol (October 2001) 0 / 50 0 / 50 0 / 50

Tetralin (June 2003) 0 /50 0 /50 0 /50

Total (%) 0/3 50 2 /3 50 ( 0.6 %) 0 /3 50

Mean 

### ± standard deviation 0.6 .% 

### ± 1. 0%

Range 0%

- 2%



### Overall Historical Incidence: All Routes

Total (%) 5 /1,3 50 ( 0.4 %) 8 /1,3 50 ( 0.6 %) 3 /1,3 50 ( 0. 2 %)

Mean 

### ± standard deviation 0.4 % 

### ± 0.8 % 0.6 % 

### ± 1. 2% 0. 2 % ± 0.6%

Range 0% -2% 0% -4% 0% -2%



### Basal Cell Carcinoma Squamous Cell Papilloma,

Keratoacanthoma, Basal Cell Adenoma,

or Basal Cell Carcinoma

Historical Incidence: Inhalation Studies

1- Bromopropane (July 2003) 0 /50 1 /50

Cumene (June 2001) 0 /50 0 /50

Diethylamine (August 2003) 0/50 1 /50

Methyl isobutyl ketone (May 2000) 0/50 0/50

α

- Methylstyrene (August 2001) 0 /50 0 /50

Propargyl alcohol (October 2001) 0 / 50 0 / 50

Tetralin (June 2003) 0 /50 0 /50

Total (%) 0 /3 50 2 /3 50 ( 0.6 %)

Mean 

### ± standard deviation 0.6 .% 

### ± 1.0 %

Range 0%

- 2%



### Overall Historical Incidence: All Routes

Total (%) 1 / 1,3 50 ( 0.1 %) 16 / 1,3 50 ( 1. 2 %)

Mean 

### ± standard deviation 0.1 % 

### ± 0.4 % 1.2 % 

### ± 1.8 %

Range 0% -2% 0% -6%

a Data as of April 29, 2009

### 114 1- Bromopropane, NTP TR 564

T ABLE 

### B4

Summary of the Incidence of Nonneoplastic Lesions in Fem ale Rats

in the 2

- Year Inhalation Study of 1- Bromopropane a



### Chamber Control 125 ppm 250 ppm 500 ppm

Disposition Summary

Animals initially in study 50 50 50 50

Early deaths

Accidental deaths 2

Moribund 13 17 17 23

Natural deaths 3 3 1

Survivors

Terminal sacrifice 34 33 30 24

Animals examined microscopically 50 50 50 50



### A limentary S ystem

Intestine l arge, c olon (47) (50) (48) (49)

Intestine l arge, r ectum (47) (50) (48) (49)

Inflammation, s uppurative 1 (2%)

Necrosis 1 (2%)

Intestine s mall, i leum (46) (50) (47) (48)

Intestine s mall, j ejunum (47) (50) (47) (48)

Liver (50) (50) (50) (50)

Angiectasis 3 (6%) 2 (4%) 2 (4%) 1 (2%)

Basophilic f ocus 40 (80%) 43 (86%) 42 (84%) 39 (78%)

Clear c ell f ocus 30 (60%) 34 (68%) 35 (70%) 35 (70%)

Cyst 1 (2%)

Degeneration, c ystic 1 (2%)

Eosinophilic f ocus 1 (2%) 1 (2%)

Hepatodiaphragmatic n odule 4 (8%) 9 (18%) 5 (10%) 5 (10%)

Mixed c ell f ocus 2 (4%) 2 (4%) 3 (6%) 3 (6%)

Necrosis 1 (2%) 1 (2%)

Vacuolization c ytoplasmic 4 0 (80%) 41 (82%) 39 (78%) 47 (94%)

Bile d uct, c yst 1 (2%)

Serosa, f ibrosis 1 (2%)

Mesentery (13) (26) (17) (23)

Necrosis 13 (100%) 26 (100%) 16 (94%) 22 (96%)

Oral m ucosa (2) (0) (1) (1)

Foreign b ody 1 (100%) 1 (100%)

Hyperplasia, s quamous 2 (100%) 1 (100%) 1 (100%)

Inflammation, chronic a ctive 1 (100%)

Pancreas (50) (50) (50) (50)

Acinus, a trophy 9 (18%) 12 (24%) 7 (14%) 7 (14%)

Salivary g lands (50) (50) (50) (50)

Hyperplasia 1 (2%)

Inflammation, c hronic a ctive 1 (2%)

Necrosis 1 (2%)

Stomach, f orestomach (50) (50) (50) (50)

Hyperplasia, b asal c ell 1 (2%) 1 (2%)

Hyperplasia, s quamous 4 (8%) 2 (4%) 2 (4%)

Ulcer 2 (4%) 3 (6%) 3 (6%) 6 (12%)

Stomach, g landular (50) (50) (50) (50)

Erosion 1 (2%)

Ulcer 3 (6%) 1 (2%) 1 (2%) 1 (2%)

Tongue (0) (1) (0) (1)

Hyperplasia, s quamous 1 (100%)

Tooth (0) (0) (1) (0)

Inflammation, c hronic a ctive 1 (100%)

a Number of animals examined microscopically at the site and the number of animals with lesion

### 1- Bromopropane, NTP TR 564 115

T ABLE 

### B4

Summary of the Incidence of Nonneoplastic Lesions in Fem ale Rats

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Cardiovascular System

Heart (50) (50) (50) (50)

Cardiomyopathy 35 (70%) 32 ( 64%) 34 (68%) 21 (42%)

Atrium, t hrombosis 1 (2%)



### Endocrine System

Adrenal c ortex (50) (50) (50) (50)

Degeneration, c ystic 1 (2%) 1 (2%)

Hyperplasia 27 (54%) 30 (60%) 24 (48%) 26 (52%)

Necrosis 1 (2%)

Vacuolization c ytoplasmic 1 2 ( 24%) 16 (32%) 16 (32%) 11 (22%)

Adrenal m edulla (50) (50) (50) (50)

Hyperplasia 4 (8%) 9 (18%) 7 (14%) 6 (12%)

Metaplasia, o sseous 1 (2%)

Islets, p ancreatic (50) (50) (50) (50)

Hyperplasia 3 (6%) 4 (8%) 4 (8%)

Metaplasia

, h epatocyte 2 (4%) 1 (2%)

Pituitary g land (50) (50) (50) (50)

Cyst 2 (4%) 1 (2%) 2 (4%) 2 (4%)

Hemorrhage 2 (4%) 4 (8%) 5 (10%) 3 (6%)

Pars d istalis, h yperplasia 7 (14%) 11 (22%) 7 (14%) 7 (14%)

Thyroid g land (50) (50) (50) (50)

Ultimobranchial c yst 1 (2%) 1 (2%)

C

- cell, h yperplasia 39 (78%) 36 (72%) 37 (74%) 37 (74%)

Follicular c ell, h yperplasia 1 (2%) 2 (4%)



### General Body System

None



### Genital System

Clitoral g land (49) (50) (49) (49)

Hyperplasia 3 (6%) 5 (10%) 9 (18%) 4 (8%)

Inflammation, c hronic a ctive 22 (45%) 2 6 (52%) 26 (53%) 18 (37%)

Ovary (50) (50) (50) (50)

Cyst 3 (6%) 1 (2%) 7 (14%) 3 (6%)

Interstitial c ell, h yperplasia 1 (2%)

Uterus (50) (50) (50) (50)

Congestion 1 (2%)

Cyst 1 (2%)

Decidual r eaction 2 (4%)

Hemorrhage 2 (4%) 1 (2%)

Hydrometra 1 (2%)

Inflammation, c hronic a ctive 1 (2%)

Endometrium, h yperplasia 1 (2%)

Vagina (3) (0) (0) (0)

Muscularis, h ypertrophy 2 (67%)

### 116 1- Bromopropane, NTP TR 564

T ABLE 

### B4

Summary of the Incidence of Nonneoplastic Lesions in Fem ale Rats

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

Hematopoietic System

Bone m arrow (50) (50) (50) (50)

Hyperplasia, h istiocytic 1 (2%) 1 (2%) 1 (2%) 1 (2%)

Lymph n ode (1) (3) (2) (2)

Inflammation, c hronic a ctive 1 (100%)

Deep c ervical, e ctasia 1 (33%)

Deep c ervical, h yperplasia, l ymphoid 1 (33%)

Pancreatic, h emorrhage 1 (50%)

Lymph n ode, b ronchial (1) (4) (5) (7)

Angiectasis 1 (100%) 2 (40%) 1 (14%)

Hemorrhage 1 (20%) 1 (14%)

Lymph n ode, m ediastinal (34) (30) (31) (28)

Angiectasis 2 (6%) 2 (7%)

Hemorrhage 2 (7%)

Lymph n ode, m esenteric (49) (49) (50) (50)

Hemorrhage 1 (2%)

Hyperplasia, l ymphoid 1 (2%)

Infiltration c ellular, h istiocyte 1 (2%) 1 (2%)

Spleen (50) (50) (50) (50)

Hematopoietic c ell p roliferation 3 3 (66%) 32 (64%) 29 (58%) 21 (42%)

Hemorrhage, c hronic 1 (2%) 1 (2%) 2 (4%)

Infarct, c hronic 1 (2%) 1 (2%) 1 (2%) 1 (2%)

Inflammation, g ranulomatous 2 (4%)

Lymphoid f ollicle, a trophy 1 (2%)

Thymus (46) (45) (45) (43)

Cyst 1 (2%)



### Integumentary System

Mammary g land (50) (50) (50) (50)

Galactocele 1 (2%) 3 (6%) 1 (2%)

Hyperplasia 1 (2%)

Inflammation, c hronic a ctive 1 (2%) 1 (2%)

Skin (50) (50) (50) (50)

Hyperkeratosis 1 (2%) 2 (4%) 2 (4%)

Hyperplasia, s quamous 1 (2%)

Inflammation, s uppurative, c hronic 1 (2%) 1 (2%)

Inflammation, c hronic a ctive 1 (2%) 2 (4%) 1 (2%)

Ulcer 1 (2%) 1 (2%)



### Musculoskeletal System

Bone (50) (50) (50) (50)

Fracture 1 (2%)

Inflammation, s uppurative, c hronic 1 (2%)

Skeletal m uscle (0) (0) (1) (4)



### Nervous System

Brain (50) (50) (50) (50)

Angiectasis 1 (2%)

Hemorrhage 1 (2%)

Spinal c ord (0) (1) (0) (3)

Cyst e pithelial i nclusion 1 (33%)

### 1- Bromopropane, NTP TR 564 117

T ABLE 

### B4

Summary of the Incidence of Nonneoplastic Lesions in Fem ale Rats

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 125 ppm 250 ppm 500 ppm

R espiratory S ystem

Larynx (50) (50) (50) (50)

Foreign b ody 8 (16%) 6 (12%) 3 (6%) 4 (8%)

Inflammation, s uppurative, c hronic 3 (6%)

Inflammation, c hronic a ctive 18 (36%) 25 (50%) 30 (60%) 32 (64%)

Metaplasia, s quamous 3 (6%) 2 (4%) 6 (12%) 21 (42%)

Necrosis 1 (2%)

Respiratory e pithelium, h yperplasia 1 (2%)

Lung (50) (50) (50) (50)

Hemorrhage 4 (8%) 6 (12%) 4 (8%) 4 (8%)

Inflammation, s uppurative, c hronic 4 (8%)

Inflammation, c hronic a ctive 6 (12%) 10 (20%) 12 (24%) 2 (4%)

Alveolar e pithelium, h yperplasia 1 1 (22%) 10 (20%) 16 (32%) 8 (16%)

Alveolus, i nfiltration c ellular, h istiocyte 30 (60%) 30 (60%) 29 (58%) 26 (52%)

Nose (50) (50) (49) (50)

Foreign b ody 6 (12%) 6 (12%) 6 (12%) 9 (18%)

Inflammation, s uppurative, c hronic 1 (2%) 3 (6%) 7 (14%)

Inflammation, c hronic a ctive 24 (48%) 37 (74%) 37 (76%) 36 (72%)

Epithelium, a ccumulation, h yaline d roplet 48 (96%) 48 (96%) 48 (98%) 47 (94%)

Glands, h yperplasia 6 (12%) 23 (46%) 28 (57%) 30 (60%)

Olfactory e pithelium, h yperplasia 1 (2%)

Olfactory e pithelium, m etaplasia,

r espiratory 3 (6%) 4 (8%) 6 (12%) 9 (18%)

Respiratory e pithelium, h yperplasia 5 (10%) 13 (26%) 9 (18%) 18 (36%)

Respiratory e pithelium, m etaplasia,

squamous 1 (2%) 1 (2%) 5 (10%) 1 (2%)

Pleura (0) (0) (1) (0)

Trachea (50) (50) (50) (50)

Inflammation, c hronic a ctive 4 (8%) 1 (2%) 6 (12%)

Epithelium, h yperplasia 4 (8%)

Epithelium, m etaplasia, s quamous 1 (2%)

Epithelium, n ecrosis 1 (2%)



### Special Senses System

Ear (0) (0) (1) (1)

Inflammation, s uppurative, c hronic 1 (100%)

Eye (50) (50) (50) (50)

Cornea, i nflammation, c hronic a ctive 1 (2%)

Lens, c ataract 4 (8%) 3 (6%) 5 (10%) 2 (4%)

Harderian g land (50) (50) (50) (50)

Inflammation, s uppurative, c hronic 1 (2%)

Inflammation, c hronic a ctive 1 (2%) 1 (2%)

Zymbal ’ s g land (1) (1) (2) (0)



### Urinary System

Kidney (49) (50) (50) (50)

Cyst 1 (2%) 1 (2%)

Nephropathy, c hronic 35 (71%) 35 (70%) 31 (62%) 29 (58%)

Cortex, i nfarct 1 (2%)

Cortex, r enal t ubule, a ccumulation,

h yaline droplet 1 (2%)

Pelvis, i nflammation, s uppurative 1 (2%)

Pelvis, i nflammation, c hronic a ctive 1 (2%)

### 118 1- Bromopropane, NTP TR 564

### 119



# APPENDIX C

SUMMARY OF LESIONS I N MALE MICE

IN THE 2

- YEAR INHALATION STUDY

OF 1- BROMOPROPANE

T ABLE 

### C1 Summary of the Incidence of Neoplasms in Male Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

....................................................................... 120

T ABLE 

### C2 Statistical Analysis of Primary Neoplasms in Male Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

...................................................................... 125

T ABLE 

### C3 Summary of the Incidence of Nonneoplastic Lesions in Male Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

...................................................................... 128

### 120 1- Bromopropane, NTP TR 564

T ABLE 

### C1

Summary of the Incidence of Ne oplas ms in M ale Mice in the 2

- Year Inhalation Study of 1- Bromopropane a



### Chamber Control 62.5 ppm 125 ppm 250 ppm

Disposition Summary

Animals initially in study 50 50 50 50

Early deaths

Moribund 6 13 9 9

Natural deaths 7 4 9 5

Survivors

Terminal sacrifice 37 33 32 36

Animals examined microscopically 50 50 50 50



### Alimentary System

Gallbladder (40) (40) (34) (38)

Intestine l arge, r ectum (45) (48) (43) (48)

Intestine s mall, d uodenum (44) (48) (43) (46)

Polyp a denomatous 1 (2%) 1 (2%)

Intestine s mall, j ejunum (44) (48) (43) (46)

Carcinoma 4 (9%) 1 (2%) 1 (2%)

Liver (50) (50) (50) (50)

Hemangiosarcoma 1 (2%) 1 (2%) 1 (2%) 1 (2%)

Hepatoblastoma 1 (2%) 1 (2%) 1 (2%)

Hepatocellular a denoma 16 (32%) 14 (28%) 19 (38%) 15 (30%)

Hepatocellular a denoma, m ultiple 14 (28%) 18 (36%) 14 (28%) 8 (16%)

Hepatocellular c arcinoma 9 (18%) 8 (16%) 15 (30%) 7 (14%)

Hepatocellular c arcinoma, m ultiple 6 (12%) 7 (14%) 4 (8%) 4 (8%)

Hepatocholangiocarcinoma 1 (2%) 1 (2%) 1 (2%) 1 (2%)

Hepatocholangiocarcinoma, m ultiple 1 (2%)

Capsule, c arcinoma, m etastatic, pancreas 1 (2%)

Mesentery (9) (11) (6) (7)

Carcinoma, m etastatic, p ancreas 1 (11%) 1 (17%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (14%)

Pancreas (48) (50) (48) (50)

Carcinoma 1 (2%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Acinar c ell, c arcinoma 1 (2%)

Salivary g lands (49) (50) (49) (50)

Stomach, f orestomach (47) (50) (48) (50)

Squamous c ell p apilloma 2 (4%)

Stomach, g landular (47) (49) (47) (49)

Tooth (8) (8) (9) (11)



### C ardiovascular S ystem

Blood v essel (3) (1) (0) (0)

Heart (50) (50) (50) (50)

Alveolar/ b ronchiolar c arcinoma, metastatic,

lung 1 (2%)

Carcinoma, m etastatic, p ancreas 1 (2%)

Hemangiosarcoma 1 (2%)

Hepatocellular c arcinoma, m etastatic, liver 1 (2%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%) 1 (2%) 2 (4%)

### 1- Bromopropane, NTP TR 564 121

T ABLE 

### C1

Summary of the Incidence of Ne oplas ms in M ale Mice in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

Endocrine System

Adrenal c ortex (49) (50) (49) (50)

Adenoma 1 (2%) 1 (2%)

Carcinoma, m etastatic, p ancreas 1 (2%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Subcapsular, a denoma 3 (6%) 3 (6%) 1 (2%)

Adrenal m edulla (50) (49) (49) (50)

Pheochromocytoma b enign 1 (2%) 1 (2%)

Pheochromocytoma m alignant 1 (2%)

Islets, p ancreatic (48) (50) (48) (50)

Adenoma 1 (2%) 1 (2%)

Pituitary g land (46) (48) (46) (48)

Pars d istalis, a denoma 1 (2%) 1 (2%)

Thyroid g land (49) (50) (48) (50)



### G eneral B ody S ystem

Peritoneum (0) (0) (1) (0)

Carcinoma, m etastatic, p ancreas 1 (100%)



### G enital S ystem

Coagulating g land (1) (0) (0) (0)

Epididymis (50) (50) (50) (50)

Preputial g land (49) (49) (50) (50)

Bilateral, s quamous c ell c arcinoma 1 (2%)

Prostate (49) (50) (48) (50)

Seminal v esicle (48) (50) (49) (50)

Adenoma 1 (2%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Testes (50) (50) (50) (50)

Interstitial c ell, a denoma 1 (2%)



### H ematopoietic S ystem

Bone m arrow (48) (48) (48) (50)

Hemangiosarcoma 1 (2%) 1 (2%)

Mast c ell t umor b enign 1 (2%)

Lymph n ode (1) (1) (3) (0)

Alveolar/ b ronchiolar c arcinoma,

metastatic, lung 1 (33%)

Lumbar, c arcinoma, m etastatic,

uncertain primary site 1 (33%)

Popliteal, c arcinoma, m etastatic,

uncertain primary site 1 (33%)

Lymph n ode, b ronchial (23) (30) (32) (31)

Carcinoma, m etastatic, p ancreas 1 (3%)

Hepatocellular c arcinoma, m etastatic, liver 1 (3%) 1 (3%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (3%) 1 (3%) 2 (6%)

Lymph n ode, m andibular (27) (28) (21) (26)

### 122 1- Bromopropane, NTP TR 564

T ABLE 

### C1

Summary of the Incidence of Ne oplas ms in M ale Mice in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

Hematopoietic System (continued)

Lymph n ode, m ediastinal (35) (32) (35) (34)

Carcinoma, m etastatic, p ancreas 1 (3%) 1 (3%)

Hepatocellular c arcinoma, m etastatic, liver 1 (3%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (3%) 1 (3%) 1 (3%) 2 (6%)

Lymph n ode, m esenteric (48) (46) (44) (48)

Carcinoma, m etastatic, p ancreas 1 (2%) 1 (2%)

Hepatocellular c arcinoma, m etastatic,

liver 1 (2%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Spleen (48) (50) (49) (50)

Hemangiosarcoma 2 (4%) 1 (2%)

Mast c ell t umor m alignant, m etastatic,

uncertain primary site 1 (2%)

Thymus (38) (35) (37) (32)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (3%) 1 (3%)



### I ntegumentary S ystem

Skin (50) (50) (50) (50)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Subcutaneous t issue, f ibrous histiocytoma 1 (2%)

Subcutaneous t issue, h emangiosarcoma 1 (2%) 1 (2%)

Subcutaneous t issue, s arcoma 1 (2%)



### M usculoskeletal S ystem

Bone (50) (49) (50) (50)

Hemangiosarcoma 1 (2%)

Osteosarcoma 1 (2%)

Skeletal m uscle (0) (2) (1) (2)

Alveolar/ b ronchiolar c arcinoma, metastatic,

lung 1 (100%)

Hemangiosarcoma 1 (50%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (50%) 1 (50%)



### N ervous S ystem

Brain (50) (50) (49) (50)

### 1- Bromopropane, NTP TR 564 123

T ABLE 

### C1

Summary of the Incidence of Ne oplas ms in M ale Mice in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

R espiratory S ystem

Larynx (48) (50) (48) (50)

Lung (50) (50) (49) (49)

Alveolar/ b ronchiolar a denoma 6 (12%) 4 (8%) 4 (8%) 4 (8%)

Alveolar/ b ronchiolar a denoma, m ultiple 1 (2%) 1 (2%)

Alveolar/ b ronchiolar c arcinoma 8 (16%) 7 (14%) 8 (16%) 9 (18%)

Alveolar/ b ronchiolar c arcinoma, m ultiple 2 (4%) 1 (2%)

Carcinoma, m etastatic, h arderian g land 1 (2%)

Carcinoma, m etastatic, p ancreas 1 (2%) 1 (2%)

Hemangiosarcoma, m etastatic, l iver 1 (2%)

Hemangiosarcoma, m etastatic, s pleen 1 ( 2%)

Hepatocellular c arcinoma, m etastatic, liver 7 (14%) 10 (20%) 12 (24%) 4 (8%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%) 1 (2%) 1 (2%) 2 (4%)

Osteosarcoma, m etastatic,

u ncertain primary site 1 (2%)

Bronchiole, a denoma 1 (2%)

Nose (50) (50) (50) (50)

Carcinoma, m etastatic, h arderian g land 1 (2%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Pleura (0) (0) (1) (1)

Alveolar/ b ronchiolar c arcinoma,

metastatic, lung 1 (100%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (100%)

Trachea (49) (50) (47) (50)



### Special Senses System

Eye (45) (47) (44) (47)

Harderian g land (46) (49) (49) (48)

Adenoma 6 (13%) 3 (6%) 2 (4%) 3 (6%)

Carcinoma 3 (7%) 1 (2%) 7 (14%) 5 (10%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Bilateral, a denoma 1 (2%)



### Urinary System

Kidney (49) (50) (50) (49)

Alveolar/ b ronchiolar c arcinoma,

metastatic, lung 1 (2%)

Hepatocellular c arcinoma, m etastatic, liver 1 (2%) 1 (2%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%) 1 (2%) 1 (2%) 2 (4%)

Urethra (0) (0) (0) (1)

Urinary b ladder (47) (50) (47) (48)



### Systemic Lesions

Multiple o rgans b (50) (50) ( 50) (50)

Histiocytic s arcoma 2 (4%) 1 (2%)

Lymphoma m alignant 2 (4%) 3 (6%)

### 124 1- Bromopropane, NTP TR 564

T ABLE 

### C1

Summary of the Incidence of Ne oplas ms in M ale Mice in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

Neoplasm Summary

Total a nimals with p rimary n eoplasms c 47 47 46 36

Total p rimary n eoplasms 90 81 90 69

Total a nimals with b enign n eoplasms 36 35 35 29

Total b enign n eoplasms 51 47 43 35

Total a nimals with m alignant neoplasms 30 29 34 24

Total m alignant n eoplasms 39 34 47 34

Total a nimals with m etastatic neoplasms 9 12 17 6

Total m etastatic n eoplasms 18 24 39 21

Total a nimals with m alignant neoplasms

of uncertain primary site 2 1

a Number of animals examined microscopically at the site and the number of animals with neoplasm

b Number of animals with any tissue examined microscopically

c Primary neoplasms: all neoplasms except metastatic neoplasms

### 1- Bromopropane, NTP TR 564 125

T ABLE 

### C2

Statistical Analysis of Primary Neoplasms in M ale Mice in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

Adrenal Cortex : Adenoma

Overall rate a 4/49 (8%) 3/50 (6%) 2/49 (4%) 0/50 (0%)

Adjusted rate b 9.1% 6.9% 4.5% 0.0%

Terminal rate c 3/37 (8%) 2/33 (6%) 2/32 (6%) 0/36 (0%)

First incidence (days) 586 655 729 (T)

— e

Poly

- 3 test d P=0.040N P=0.502N P=0.329N P=0.066N



### Harderian Gland: Adenoma

Overall rate 7/50 (14%) 3/50 (6%) 2/50 (4%) 3/50 (6%)

Adjusted rate 15.9% 6.9% 4.4% 7.2%

Terminal rate 7/37 (19%) 1/33 (3%) 1/32 (3%) 3/36 (8%)

First incidence (days) 729 (T) 684 702 729 (T)

Poly

- 3 test P=0.133N P=0.161N P=0.072N P=0.179N



### Harderian Gland: Carcinoma

Overall rate 3/50 (6%) 1/50 (2%) 7/50 (14%) 5/50 (10%)

Adjusted rate 6.8% 2.3% 15.1% 12 .0 %

Terminal rate 3/37 (8%) 1/33 (3%) 3/32 (9%) 5/36 (14%)

First incidence (days) 729 (T) 729 (T) 604 729 (T)

Poly

- 3 test P=0.123 P=0.312N P=0.178 P=0.327



### Harderian Gland: Adenoma or Carcinoma

Overall rate 10/50 (20%) 4/50 (8%) 8/50 (16%) 8/50 (16%)

Adjusted rate 22.7% 9.2% 17.2% 19.1%

Terminal rate 10/37 (27%) 2/33 (6%) 3/32 (9%) 8/36 (22%)

First incidence (days) 729 (T) 684 604 729 (T)

Poly

- 3 test P=0.547 P=0.074N P=0.350N P=0.446N



### Small Intestine (Jejunum): Carcinoma

Overall rate 4/50 (8%) 1/50 (2%) 1/50 (2%) 0/50 (0%)

Adjusted rate 9.1% 2.3% 2.2% 0.0%

Terminal rate 4/37 (11%) 0/33 (0%) 0/32 (0%) 0/36 (0%)

First incidence (days) 729 (T) 631 682

— Poly

- 3 test P=0.034N P=0.181N P=0.170N P=0.067N



### Liver: Hepatocellular Adenoma

Overall rate 30/50 (60%) 32/50 (64%) 33/50 (66%) 23/50 (46%)

Adjusted rate 62.5% 67.6% 68.3% 53.3%

Terminal rate 23/37 (62%) 24/33 (73%) 21/32 (66%) 20/36 (56%)

First incidence (days) 328 424 537 417

Poly

- 3 test P=0.186N P=0.377 P=0.347 P=0.248N



### Liver: Hepatocellular Carcinoma

Overall rate 15/50 (30%) 15/50 (30%) 19/50 (38%) 11/50 (22%)

Adjusted rate 32.7% 33.1% 39.1% 25.4%

Terminal rate 9/37 (24%) 8/33 (24%) 8/32 (25%) 7/36 (19%)

First incidence (days) 539 520 537 470

Poly

- 3 test P=0.289N P=0.571 P=0.332 P=0.302N



### Liver: Hepatocellular Adenoma or Carcinoma

Overall rate 38 /50 ( 76 %) 38 /50 ( 76 %) 41 /50 (8 2 %) 28 /50 ( 56 %)

Adjusted rate 77.7 % 78.1 % 82.3 % 63.1 %

Terminal rate 28 /37 (7 6 %) 25 /33 (7 6 %) 24 /32 ( 7 5%) 22/ 36 ( 61 %)

First incidence (days) 328 424 537 4 17

Poly

- 3 test P=0.0 66 N P=0.5 78 P=0. 370 P=0.0 89 N

### 126 1- Bromopropane, NTP TR 564

T ABLE 

### C2

Statistical Analysis of Primary Neoplasms in M ale Mice in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

Liver: Hepatocellular Carcinoma or Hepatoblastoma

Overall rate 16 /50 ( 32 %) 16 /50 ( 32 %) 19 /50 ( 38 %) 11 /50 ( 22 %)

Adjusted rate 34.9 % 35.3 % 3 9.1 % 25.4 %

Terminal rate 10 /37 ( 27 %) 9 /33 ( 27 %) 8 /32 ( 2 5%) 7 /36 (1 9 %)

First incidence (days) 539 520 537 47 0

Poly

- 3 test P=0. 208 N P=0.5 69 P=0. 415 P=0. 230N



### Liver: Hepatocellular Adenoma, Hepatocellular Carcinoma, or Hepatoblastoma

Overall rate 38/50 (76%) 39/50 (78%) 41/50 (82%) 28/50 (56%)

Adjusted rate 77.7% 80.1% 82.3% 63.1%

Terminal rate 28/37 (76%) 26/33 (79%) 24/32 (75%) 22/36 (61%)

First incidence (days) 328 424 537 417

Poly

- 3 test P=0.055N P=0.479 P=0.370 P=0.089N



### Lung: Alveolar/bronchiolar Adenoma

Overall rate 6/50 (12%) 5/50 (10%) 4/49 (8%) 5/49 (10%)

Adjusted rate 13.3% 11.5% 9.0% 11.9%

Terminal rate 4/37 (11%) 5/33 (15%) 4/32 (13%) 4/36 (11%)

First incidence (days) 544 729 (T) 729 (T) 360

Poly

- 3 test P=0.476N P=0.530N P=0.380N P=0.554N



### Lung: Alveolar/bronchiolar Carcinoma

Overall rate 8/50 (16%) 7/50 (14%) 10/49 (20%) 10/49 (20%)

Adjusted rate 17.8% 15.9% 22.0% 24.3%

Terminal rate 6/37 (16%) 6/33 (18%) 5/32 (16%) 9/36 (25%)

First incidence (days) 539 520 617 708

Poly

- 3 test P=0.209 P=0.518N P=0.407 P=0.317



### Lung: Alveolar/bronchiolar Adenoma or Carcinoma

Overall rate 13/50 (26%) 12/50 (24%) 14/49 (29%) 15/49 (31%)

Adjusted rate 28.3% 27.3% 30.8% 35.7%

Terminal rate 9/37 (24%) 11/33 (33%) 9/32 (28%) 13/36 (36%)

First incidence (days) 539 520 617 360

Poly

- 3 test P=0.225 P=0.552N P=0.484 P=0.301



### All Organs: Hemangiosarcoma

Overall rate 2/50 (4%) 4/50 (8%) 2/50 (4%) 2/50 (4%)

Adjusted rate 4.5% 9.2% 4.4% 4.8%

Terminal rate 2/37 (5%) 3/33 (9%) 1/32 (3%) 1/36 (3%)

First incidence (days) 729 (T) 661 718 708

Poly

- 3 test P=0.477N P=0.331 P=0.684N P=0.675



### All Organs: Malignant Lymphoma

Overall rate 2/50 (4%) 0/50 (0%) 3/50 (6%) 0/50 (0%)

Adjusted rate 4.5% 0.0% 6.6% 0.0%

Terminal rate 1/37 (3%) 0/33 (0%) 3/32 (9%) 0/36 (0%)

First incidence (days) 578

— 729 (T)

— Poly

- 3 test P=0.313N P=0.244N P=0.506 P=0.252N



### All Organs: Benign Neoplasms

Overall rate 36/50 (72%) 35/50 (70%) 35/50 (70%) 29/50 (58%)

Adjusted rate 73.2% 73.5% 72.4% 65.9%

Terminal rate 26/37 (70%) 26/33 (79%) 23/32 (72%) 25/36 (69%)

First incidence (days) 328 424 537 360

Poly

- 3 test P=0.232N P=0.582 P=0.556N P=0.290N

### 1- Bromopropane, NTP TR 564 127

T ABLE 

### C2

Statistical Analysis of Primary Neoplasms in M ale Mice in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

All Organs: Malignant Neoplasms

Overall rate 30/50 (60%) 30/50 (60%) 34/50 (68%) 24/50 (48%)

Adjusted rate 62.7% 63.5% 69.0% 54.8%

Terminal rate 20/37 (54%) 18/33 (55%) 19/32 (59%) 19/36 (53%)

First incidence (days) 328 520 537 470

Poly

- 3 test P=0.270N P=0.554 P=0.329 P=0.289N



### All Organs: Benign or Malignant Neoplasms

Overall rate 47/50 (94%) 47/50 (94%) 46/50 (92%) 36/50 (72%)

Adjusted rate 94.0% 94.3% 92.4% 78.6%

Terminal rate 34/37 (92%) 31/33 (94%) 29/32 (91%) 28/36 (78%)

First incidence (days) 328 424 537 360

Poly

- 3 test P=0.005N P=0.640 P=0.529N P=0.024N

(T) Terminal sacrifice

a Number of neoplasm

- bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal

cortex, liver, and lung; for other tissues, denominator is number of animals necropsied.

b Poly

- 3 estimated neoplasm incidence after adjustment for intercurrent mortality

c Observed incidence at terminal kill

d Beneath the chamber control incidence is the P value associated with the trend test. Beneath the exposed group incidence are the P va lues

corresponding to pairwise comparisons between the chamber controls and that expose d group. The Poly

- 3 test accounts for the differential

mortality in animals that do not reach terminal sacrifice. A negative trend or a lower incidence in an expos ure group is indicated by N

.

e Not applicable; no neoplasms in animal group

### 128 1- Bromopropane, NTP TR 564

T ABLE 

### C3

Summary of the Incidence of Nonneoplastic Lesions in M ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane a



### Chamber Control 62.5 ppm 125 ppm 250 ppm

Disposition Summary

Animals initially in study 50 50 50 50

Early deaths

Moribund 6 13 9 9

Natural deaths 7 4 9 5

Survivors

Terminal sacrifice 37 33 32 36

Animals examined microscopically 50 50 50 50



### Alimentary System

Gallbladder (40) (40) (34) (38)

Degeneration, h yaline 2 (5%) 1 (3%)

Hemorrhage 1 (3%)

Hyperplasia 1 (3%)

Infiltration c ellular, p olymorphonuclear 1 (3%)

Intestine l arge, r ectum (45) (48) (43) (48)

Inflammation, s uppurative 1 (2%)

Ulcer 1 (2%)

Artery, i nflammation, c hronic a ctive 1 (2%)

Intestine s mall, d uodenum (44) (48) (43) (46)

Inflammation, a cute 1 (2%)

Intestine s mall, j ejunum (44) (48) (43) (46)

Liver (50) (50) (50) (50)

Angiectasis 3 (6%)

Basophilic f ocus 1 (2%) 2 (4%) 1 (2%) 3 (6%)

Clear c ell f ocus 9 (18%) 14 (28%) 16 (32%) 12 (24%)

Eosinophilic f ocus 20 (40%) 16 (32%) 8 (16%) 9 (18%)

Fatty c hange, f ocal 1 (2%) 3 (6%) 5 (10%)

Inflammation, c hronic 1 (2%) 1 (2%)

Mineralization 1 (2%)

Mixed c ell f ocus 2 (4%) 7 (14%) 1 (2%) 1 (2%)

Necrosis 5 (10%) 4 (8%) 3 (6%) 9 (18%)

Tension l ipidosis 4 (8%) 3 (6%) 6 (12%) 2 (4%)

Vacuolization c ytoplasmic 1 (2%)

Bile d uct, h yperplasia 1 (2%)

Mesentery (9) (11) (6) (7)

Artery, i nflammation 1 (14%)

Artery, m ineralization 1 (11%)

Fat, n ecrosis 7 (78%) 11 (100%) 5 (83%) 5 (71%)

Pancreas (48) (50) (48) (50)

Atrophy 1 (2%) 1 (2%)

Basophilic f ocus 1 (2%) 1 (2%)

Cyst 1 (2%)

Inflammation, c hronic a ctive 1 (2%)

Salivary g lands (49) (50) (49) (50)

Atrophy 1 (2%) 1 (2%)

Stomach, f orestomach (47) (50) (48) (50)

Hyperplasia, s quamous 5 (11%) 7 (14%) 5 (10%) 3 (6%)

Inflammation 2 (4%) 2 (4%) 1 (2%)

Ulcer 2 (4%) 4 (8%) 3 (6%) 2 (4%)

Artery, i nflammation, c hronic a ctive 1 (2%)

a Number of animals examined microscopically at the site and the number of animals with lesion

### 1- Bromopropane, NTP TR 564 129

T ABLE 

### C3

Summary of the Incidence of Nonneoplastic Lesions in M ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

Alimentary System (continued)

Stomach, g landular (47) (49) (47) (49)

Mineralization 2 (4%)

Artery, i nflammation, c hronic a ctive 1 (2%)

Tooth (8) (8) (9) (11)

Dysplasia 7 (88%) 8 (100%) 9 (100%) 11 (100%)

Malformation 1 (13%)



### Cardiovascular System

Blood v essel (3) (1) (0) (0)

Inflammation, c hronic a ctive 1 (33%)

Thrombosis 1 (100%)

Aorta, m ineralization 2 (67%)

Heart (50) (50) (50) (50)

Cardiomyopathy 12 (24%) 10 (20%) 14 (28%) 9 (18%)

Hemorrhage 1 (2%)

Inflammation, a cute 1 (2%)

Mineralization 2 (4%)

Thrombosis 1 (2%) 2 (4%) 1 (2%)

Artery, i nflammation, c hronic a ctive 1 (2%) 1 (2%)

Epicardium, i nflammation, c hronic 1 (2%)



### Endocrine System

Adrenal c ortex (49) (50) (49) (50)

Hyperplasia 16 (33%) 18 (36%) 14 (29%) 12 (24%)

Hypertrophy 23 (47%) 19 (38%) 22 (45%) 28 (56%)

Subcapsular, h yperplasia 1 (2%)

Adrenal m edulla (50) (49) (49) (50)

Hyperplasia 1 (2%) 2 (4%) 1 (2%) 2 (4%)

Islets, p ancreatic (48) (50) (48) (50)

Hyperplasia 1 (2%)

Pituitary g land (46) (48) (46) (48)

Cyst 1 (2%) 1 (2%)

Pars d istalis, h yperplasia 2 (4%) 2 (4%) 1 (2%) 1 (2%)

Pars i ntermedia, h yperplasia 1 (2%)

Thyroid g land (49) (50) (48) (50)

Follicular c ell, h yperplasia 1 (2%)



### General Body System

Peritoneum (0) (0) (1) (0)



### Genital System

Coagulating g land (1) (0) (0) (0)

Inflammation, s uppurative 1 (100%)

Epididymis (50) (50) (50) (50)

Granuloma s perm 1 (2%) 1 (2%) 1 (2%)

Necrosis 2 (4%)

Preputial g land (49) (49) (50) (50)

Inflammation, s uppurative 1 (2%)

Inflammation, g ranulomatous 1 (2%) 1 (2%)

### 130 1- Bromopropane, NTP TR 564

T ABLE 

### C3

Summary of the Incidence of Nonneoplastic Lesions in M ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

Genital System (continued)

Prostate (49) (50) (48) (50)

Inflammation, s uppurative 2 (4%) 2 (4%) 2 (4%)

Inflammation, c hronic a ctive 1 (2%) 1 (2%)

Artery, i nflammation, c hronic a ctive 1 (2%)

Seminal v esicle (48) (50) (49) (50)

Amyloid d eposition 1 (2%)

Inflammation, s uppurative 1 (2%)

Inflammation, c hronic a ctive 1 (2%)

Testes (50) (50) (50) (50)

Atrophy 1 (2%) 1 (2%) 1 (2%) 3 (6%)



### Hematopoietic System

Bone m arrow (48) (48) (48) (50)

Lymph n ode (1) (1) (3) (0)

Lymph n ode, b ronchial (23) (30) (32) (31)

Infiltration c ellular, m ixed c ell 1 (3%)

Lymph n ode, m andibular (27) (28) (21) (26)

Lymph n ode, m ediastinal (35) (32) (35) (34)

Infiltration c ellular, m ixed c ell 1 (3%)

Lymph n ode, m esenteric (48) (46) (44) (48)

Angiectasis 1 (2%)

Spleen (48) (50) (49) (50)

Angiectasis 1 (2%)

Atrophy 7 (15%) 8 (16%) 2 (4%) 3 (6%)

Congestion 4 (8%) 4 (8%) 1 (2%) 3 (6%)

Hematopoietic c ell p roliferation 20 (42%) 19 (38%) 14 (29%) 16 (32%)

Hyperplasia, l ymphoid 5 (10%) 2 (4%) 5 (10%) 2 (4%)

Infarct 1 (2%)

Necrosis 1 (2%)

Necrosis, l ymphoid 1 (2%) 1 (2%) 2 (4%)

Thymus (38) (35) (37) (32)

Atrophy 13 (34%) 12 (34%) 19 (51%) 12 (38%)

Cyst 5 (13%) 1 (3%) 7 (19%) 4 (13%)

Necrosis, l ymphoid 1 (3%) 1 (3%) 3 (9%)

Medulla, h yperplasia, l ymphoid 16 (42%) 9 (26%) 14 (38%) 6 (19%)



### Integumentary System

Skin (50) (50) (50) (50)

Cyst e pithelial i nclusion 1 (2%)

Inflammation, c hronic a ctive 5 (10%) 5 (10%) 3 (6%) 8 (16%)

Ulcer 1 (2%)



### Musculoskeletal System

Bone (50) (49) (50) (50)

Hyperostosis 2 (4%) 1 (2%)

Necrosis 1 (2%)

Skeletal m uscle (0) (2) (1) (2)



### Nervous System

Brain (50) (50) (49) (50)

### 1- Bromopropane, NTP TR 564 131

T ABLE 

### C3

Summary of the Incidence of Nonneoplastic Lesions in M ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

R espiratory S ystem

Larynx (48) (50) (48) (50)

Vacuolization c ytoplasmic 5 (10%) 10 (21%) 11 (22%)

Lung (50) (50) (49) (49)

Congestion, c hronic 1 (2%) 1 (2%)

Hemorrhage 2 (4%) 2 (4%) 1 (2%)

Inflammation, c hronic a ctive 2 (4%)

Mineralization 1 (2%)

Pigmentation 1 (2%)

Thrombosis 1 (2%)

Alveolar e pithelium, h yperplasia 2 (4%) 2 (4%) 1 (2%) 5 (10%)

Alveolus, i nfiltration c ellular, h istiocyte 3 (6%) 1 (2%) 1 (2%)

Bronchiole, h yperplasia 3 (6%) 1 (2%)

Bronchiole, n ecrosis 1 (2%)

Bronchiole, r egeneration 1 (2%) 44 (88%) 38 (78%) 47 (96%)

Bronchiole, v acuolization c ytoplasmic 18 (36%) 19 (39%) 17 (35%)

Nose (50) (50) (50) (50)

Foreign b ody 1 (2%)

Inflammation, s uppurative 2 (4%) 3 (6%) 2 (4%) 3 (6%)

Polyp, i nflammatory 3 (6%)

Glands, h yperplasia 1 (2%)

Olfactory e pithelium, a trophy 2 (4%) 4 (8%) 7 (14%) 4 (8%)

Olfactory e pithelium, m etaplasia,

r espiratory 7 (14%) 6 (12%) 3 (6%)

Respiratory e pithelium, h yperplasia 16 (32%) 29 (58%) 23 (46%) 26 (52%)

Respiratory e pithelium, v acuolization

cytoplasmic 12 (24%) 19 (38%) 20 (40%)

Pleura (0) (0) (1) (1)

Trachea (49) (50) (47) (50)

Vacuolization c ytoplasmic 15 (30%) 24 (51%) 24 (48%)



### S pecial S enses S ystem

Eye (45) (47) (44) (47)

Cataract 1 (2%) 1 (2%) 1 (2%)

Degeneration 1 (2%) 1 (2%) 1 (2%)

Cornea, i nflammation, a cute 1 (2%)

Cornea, i nflammation, c hronic a ctive 2 (4%) 1 (2%)

Retina, a trophy 1 (2%) 1 (2%)

Harderian g land (46) (49) (49) (48)

Hyperplasia 3 (7%) 4 (8%) 1 (2%) 4 (8%)



### U rinary S ystem

Kidney (49) (50) (50) (49)

Amyloid d eposition 1 (2%)

Cyst 1 (2%)

Hydronephrosis 1 (2%) 1 (2%)

Infarct 1 (2%) 2 (4%) 3 (6%) 1 (2%)

Inflammation, s uppurative 1 (2%) 2 (4%) 2 (4%)

Metaplasia, o sseous 2 (4%) 1 (2%)

Mineralization 2 (4%)

Nephropathy 44 (90%) 46 (92%) 47 (94%) 41 (84%)

Papilla, n ecrosis 1 (2%)

Renal t ubule, h yperplasia 2 (4%) 1 (2%) 2 (4%)

Renal t ubule, n ecrosis 1 (2%)

### 132 1- Bromopropane, NTP TR 564

T ABLE 

### C3

Summary of the Incidence of Nonneoplastic Lesions in M ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

Urinary System (continued)

Urethra (0) (0) (0) (1)

Bulbourethral g land, a ngiectasis 1 (100%)

Urinary b ladder (47) (50) (47) (48)

Inflammation, c hronic a ctive 1 (2%) 1 (2%) 1 (2%)

Mineralization 1 (2%)

### 133



# APPENDIX D

SUMMARY OF LESIONS IN FEMALE MICE

IN THE 2

- YEAR INHALATION STUDY

OF 1- BROMOPROPANE

T ABLE 

### D1 Summary of the Incidence of Neoplasms in Fem ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

....................................................................... 134

T ABLE 

### D2 Statistical Analysis of Primary Neoplasms in Fem ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

...................................................................... 138

T ABLE 

### D3 Historical Incidence of Alveolar/bronchiolar Neoplasms

in Control Female B6C3F1 Mice

...................................................................................................... 142

T ABLE 

### D4 Summary of the Incidence o f Nonneoplastic Lesions in Fem ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

...................................................................... 143

### 134 1- Bromopropane, NTP TR 564

T ABLE 

### D1

Summary of the Incidence of Ne oplas ms in Fem ale Mice in the 2

- Year Inhalation Study of 1- Bromopropane a



### Chamber Control 62.5 ppm 125 ppm 250 ppm

Disposition Summary

Animals initially in study 50 50 50 50

Early deaths

Moribund 10 7 8 8

Natural deaths 4 3 5

Survivors

Died last week of study 1 1

Terminal sacrifice 3 5 40 37 4 1

Animals examined microscopically 50 50 50 50



### Alimentary System

Esophagus (50) (50) (50) (50)

Squamous c ell c arcinoma 1 ( 2%)

Gallbladder (43) (36) (36) (44)

Intestine l arge, c ecum (48) (49) (46) (50)

Leiomyoma 1 (2%) 1 (2%)

Intestine l arge, c olon (48) (49) (46) (50)

Leiomyoma 1 (2%)

Intestine l arge, r ectum (47) (49) (46) (50)

Intestine s mall, d uodenum (46) (49) (46) (50)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Intestine s mall, i leum (48) (49) (46) (50)

Hemangiosarcoma 1 (2%)

Intestine s mall, j ejunum (46) (49) (46) (50)

Hemangiosarcoma 1 (2%)

Leiomyosarcoma 1 (2%)

Liver (50) (50) (50) (50)

Hemangiosarcoma 1 (2%)

Hepatoblastoma 1 (2%)

Hepatocellular a denoma 13 (26%) 8 (16%) 13 (26%) 9 (18%)

Hepatocellular a denoma, m ultiple 6 (12%) 5 (10%) 2 (4%) 3 (6%)

Hepatocellular c arcinoma 5 (10%) 4 (8%) 7 (14%)

Hepatocholangiocarcinoma 1 (2%)

Mesentery (11) (9) (11) (13)

Hemangiosarcoma 1 (9%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (8%)

Pancreas (50) (49) (50) (50)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Salivary g lands (50) (50) (50) (50)

Stomach, f orestomach (50) (50) (49) (50)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Squamous c ell p apilloma 2 (4%)

Stomach, g landular (50) (49) (48) (50)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Leiomyosarcoma, m etastatic,

i ntestine small, jejunum 1 (2%)

Tongue (0) (0) (1) (0)

Squamous c ell c arcinoma 1 (100%)

Tooth (0) (1) (0) (0)

### 1- Bromopropane, NTP TR 564 135

T ABLE 

### D1

Summary of the Incidence of Ne oplas ms in Fem ale Mice in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

C ardiovascular S ystem

Heart (50) (50) (50) (50)

Hemangiosarcoma 1 (2%)



### E ndocrine S ystem

Adrenal c ortex (50) (50) (50) (50)

Hepatocholangiocarcinoma, metastatic,

liver 1 (2%)

Adrenal m edulla (50) (50) (49) (50)

Pheochromocytoma b enign 3 (6%) 2 (4%)

Pheochromocytoma m alignant 1 (2%)

Islets, p ancreatic (50) (49) (49) (49)

Carcinoma 1 (2%)

Pituitary g land (46) (49) (47) (48)

Pars d istalis, a denoma 5 (11%) 7 (14%) 5 (11%) 9 (19%)

Pars d istalis, c arcinoma 1 (2%)

Pars i ntermedia, a denoma 4 (9%)

Thyroid g land (49) (50) (46) (50)



### G eneral B ody S ystem

Peritoneum (0) (0) (0) (1)

Leiomyosarcoma, m etastatic,

i ntestine small, jejunum 1 (100%)



### G enital S ystem

Ovary (50) (50) (50) (49)

Cystadenoma 3 (6%) 1 (2%) 1 (2%) 2 (4%)

Granulosa c ell t umor b enign 1 (2%)

Granulosa c ell t umor m alignant 1 (2%)

Luteoma 1 (2%) 1 (2%)

Uterus (50) (50) (50) (50)

Carcinoma 1 (2%)

Choriocarcinoma, m etastatic,

u ncertain primary site 1 (2%)

Leiomyosarcoma 1 (2%)

Polyp s tromal 2 (4%) 2 (4%) 3 (6%)



### H ematopoietic S ystem

Bone m arrow (50) (50) (49) (50)

Lymph n ode (6) (5) (3) (8)

Lymph n ode, b ronchial (37) (32) (39) (34)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (3%)

Lymph n ode, m andibular (45) (30) (30) (37)

Lymph n ode, m ediastinal (45) (39) (41) (37)

Hemangiosarcoma 1 (2%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (3%)

Lymph n ode, m esenteric (48) (47) (47) (49)

Spleen (50) (49) (50) (50)

Hemangiosarcoma 1 (2%) 1 (2%) 1 (2%)

Thymus (49) (45) (45) (45)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Thymoma b enign 1 (2%)

### 136 1- Bromopropane, NTP TR 564

T ABLE 

### D1

Summary of the Incidence of Ne oplas ms in Fem ale Mice in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

I ntegumentary S ystem

Mammary g land (50) (50) (50) (50)

Adenosquamous c arcinoma 1 (2%) 2 (4%)

Carcinoma 2 (4%) 1 (2%)

Skin (50) (50) (50) (50)

Sebaceous g land, c arcinoma 1 (2%)

Subcutaneous t issue, f ibrosarcoma 1 (2%)

Subcutaneous t issue, f ibrous histiocytoma 1 (2%) 2 (4%)

Subcutaneous t issue, h emangioma 1 (2%)

Subcutaneous t issue, hemangiosarcoma 1 (2%)

Subcutaneous t issue, s arcoma 7 (14%) 2 (4%) 3 (6%) 1 (2%)



### Musculoskeletal System

Bone (50) (50) (50) (50)

Hemangiosarcoma 1 (2%)

Skeletal m uscle (0) (0) (0) (1)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (100%)



### Nervous System

Brain (50) (50) (50) (50)

Carcinoma, m etastatic, p ituitary g land 1 (2%)

Meningioma b enign 1 (2%)

Peripheral n erve (1) (1) (1) (1)

Spinal c ord (1) (1) (1) (1)



### Respiratory System

Larynx (50) (50) (50) (50)

Lung (50) (50) (50) (50)

Alveolar/ b ronchiolar a denoma 1 (2%) 6 (12%) 4 (8%) 8 (16%)

Alveolar/ b ronchiolar a denoma, m ultiple 2 (4%)

Alveolar/ b ronchiolar c arcinoma 5 (10%) 4 (8%) 3 (6%)

Alveolar/ b ronchiolar c arcinoma, m ultiple 2 (4%) 1 (2%) 1 (2%)

Choriocarcinoma, m etastatic, u terus 1 (2%)

Hepatoblastoma, m etastatic, l iver 1 (2%)

Hepatocellular c arcinoma, m etastatic,

liver 1 (2%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Osteosarcoma, m etastatic,

u ncertain primary site 1 (2%)

Sarcoma, m etastatic, s kin 1 (2%)

Bronchiole, a denoma 1 (2%) 1 (2%)

Nose (50) (50) (50) (50)

Pleura (0) (0) (1) (0)

Trachea (50) (49) (50) (50)



### Special Senses System

Eye (47) (49) (45) (50)

Harderian g land (49) (49) (48) (50)

Adenoma 3 (6%) 2 (4%) 4 (8%) 2 (4%)

Carcinoma 2 (4%) 1 (2%) 1 (2%)

Zymbal ’ s g land (0) (1) (0) (0)

Carcinoma 1 (100%)

### 1- Bromopropane, NTP TR 564 137

T ABLE 

### D1

Summary of the Incidence of Ne oplas ms in Fem ale Mice in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

U rinary S ystem

Kidney (50) (49) (49) (50)

Hepatoblastoma, m etastatic, l iver 1 (2%)

Hepatocholangiocarcinoma, m etastatic,

liver 1 (2%)

Urinary b ladder (50) (49) (48) (50)

Hemangiosarcoma 1 (2%)



### S ystemic L esions

Multiple o rgans b (50) (50) (50) (50)

Histiocytic s arcoma 1 (2%) 2 (4%) 2 (4%)

Lymphoma m alignant 17 (34%) 9 (18%) 15 (30%) 11 (22%)



### Neoplasm Summary

Total a nimals with p rimary n eoplasms c 42 37 43 34

Total p rimary n eoplasms 79 66 82 67

Total a nimals with b enign n eoplasms 22 24 30 28

Total b enign n eoplasms 35 34 43 41

Total a nimals with m alignant neoplasms 33 27 32 22

Total m alignant n eoplasms 44 32 39 26

Total a nimals with m etastatic neoplasms 2 2 2 2

Total m etastatic n eoplasms 2 3 3 14

Total a nimals with m alignant neoplasms

of uncertain primary site 1 1

a Number of animals examined microscopically at the site and the number of animals with neoplasm

b Number of animals with any tissue examined microscopically

c Primary neoplasms: all neoplasms except metastatic neoplasms

### 138 1- Bromopropane, NTP TR 564

T ABLE 

### D2

Statistical Analysis of Primary Neoplasms in Fem ale Mice in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

Adrenal Medulla: Benign Pheochromocytoma

Overall rate a 3/50 (6%) 0/50 (0%) 2/49 (4%) 0/50 (0%)

Adjusted rate b 6.7% 0.0% 4.5% 0.0%

Terminal rate c 2/36 (6%) 0/40 (0%) 1/37 (3%) 0/41 (0%)

First incidence (days) 708

— e 631

— Poly

- 3 test d P=0.123N P=0.110N P=0.503N P=0.109N



### Adrenal Medulla: Benign or Malignant Pheochromocytoma

Overall rate 4/50 (8%) 0/50 (0%) 2/49 (4%) 0/50 (0%)

Adjusted rate 9.0% 0.0% 4.5% 0.0%

Terminal rate 3/36 (8%) 0/40 (0%) 1/37 (3%) 0/41 (0%)

First incidence (days) 708

— 631

— Poly

- 3 test P=0.060N P=0.054N P=0.340N P=0.054N



### Harderian Gland: Adenoma

Overall rate 3/50 (6%) 2/50 (4%) 4/50 (8%) 2/50 (4%)

Adjusted rate 6.7% 4.3% 8.9% 4.2%

Terminal rate 3/36 (8%) 2/40 (5%) 4/37 (11%) 1/41 (2%)

First incidence (days) 731 (T) 731 (T) 731 (T) 663

Poly

- 3 test P=0.456N P=0.477N P=0.505 P=0.472N



### Harderian Gland: Adenoma or Carcinoma

Overall rate 5/50 (10%) 2/50 (4%) 4/50 (8%) 3/50 (6%)

Adjusted rate 11.2% 4.3% 8.9% 6.3%

Terminal rate 5/36 (14%) 2/40 (5%) 4/37 (11%) 2/41 (5%)

First incidence (days) 731 (T) 731 (T) 731 (T) 663

Poly

- 3 test P=0.372N P=0.196N P=0.494N P=0.323N



### Liver: Hepatocellular Adenoma

Overall rate 19/50 (38%) 13/50 (26%) 15/50 (30%) 12/50 (24%)

Adjusted rate 40.8% 27.6% 32.3% 25.4%

Terminal rate 14/36 (39%) 12/40 (30%) 11/37 (30%) 11/41 (27%)

First incidence (days) 481 678 589 703

Poly

- 3 test P=0.108N P=0.127N P=0.263N P=0.083N



### Liver: Hepatocellular Carcinoma

Overall rate 5/50 (10%) 4/50 (8%) 7/50 (14%) 0/50 (0%)

Adjusted rate 11.1% 8.5% 15.4% 0.0%

Terminal rate 3/36 (8%) 3/40 (8%) 5/37 (14%) 0/41 (0%)

First incidence (days) 669 678 589

— Poly

- 3 test P=0.047N P=0.471N P=0.388 P=0.027N



### Liver: Hepatocellular Adenoma or Carcinoma

Overall rate 24/50 (48%) 15/50 (30%) 20/50 (40%) 12/50 (24%)

Adjusted rate 51.1% 31.8% 43.0% 25.4%

Terminal rate 17/36 (47%) 14/40 (35%) 15/37 (41%) 11/41 (27%)

First incidence (days) 481 678 589 703

Poly

- 3 test P=0.017N P=0.043N P=0.282N P=0.007N



### Lung: Alveolar/bronchiolar Adenoma

Overall rate 1/50 (2%) 6/50 (12%) 4/50 (8%) 10/50 (20%)

Adjusted rate 2.2% 12.8% 8.9% 20.8%

Terminal rate 1/36 (3%) 6/40 (15%) 4/37 (11%) 7/41 (17%)

First incidence (days) 731 (T) 731 (T) 731 (T) 649

Poly

- 3 test P=0.007 P=0.064 P=0.181 P=0.006

### 1- Bromopropane, NTP TR 564 139

T ABLE 

### D2

Statistical Analysis of Primary Neoplasms in Fem ale Mice in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

Lung: Alveolar/bronchiolar Carcinoma

Overall rate 0/50 (0%) 7/50 (14%) 5/50 (10%) 4/50 (8%)

Adjusted rate 0.0% 14.9% 11.1% 8.5%

Terminal rate 0/36 (0%) 6/40 (15%) 5/37 (14%) 4/41 (10%)

First incidence (days)

— 709 731 (T) 731 (T)

Poly

- 3 test P=0.277 P=0.009 P=0.031 P=0.068



### Lung: Alveolar/bronchiolar Adenoma or Carcinoma

Overall rate 1/50 (2%) 9/50 (18%) 8/50 (16%) 14/50 (28%)

Adjusted rate 2.2% 19.2% 17.8% 29.2%

Terminal rate 1/36 (3%) 8/40 (20%) 8/37 (22%) 11/41 (27%)

First incidence (days) 731 (T) 709 731 (T) 649

Poly

- 3 test P<0.001 P=0.010 P=0.016 P<0.001



### Ovary: Cystadenoma

Overall rate 3/50 (6%) 1/50 (2%) 1/50 (2%) 2/49 (4%)

Adjusted rate 6.7% 2.1% 2.2% 4.3%

Terminal rate 3/36 (8%) 1/40 (3%) 1/37 (3%) 1/40 (3%)

First incidence (days) 731 (T) 731 (T) 731 (T) 730

Poly

- 3 test P=0.480N P=0.287N P=0.302N P=0.484N



### Pituitary Gland (Pars Distalis): Adenoma

Overall rate 5/46 (11%) 7/49 (14%) 5/47 (11%) 9/48 (19%)

Adjusted rate 12.2% 15.2% 11.8% 19.1%

Terminal rate 5/34 (15%) 6/39 (15%) 5/35 (14%) 6/41 (15%)

First incidence (days) 731 (T) 723 731 (T) 470

Poly

- 3 test P=0.237 P=0.461 P=0.608N P=0.278



### Pituitary Gland (Pars Distalis): Adenoma or Carcinoma

Overall rate 6/46 (13%) 7/49 (14%) 5/47 (11%) 9/48 (19%)

Adjusted rate 14.6% 15.2% 11.8% 19.1%

Terminal rate 5/34 (15%) 6/39 (15%) 5/35 (14%) 6/41 (15%)

First incidence (days) 708 723 731 (T) 470

Poly

- 3 test P=0.322 P=0.586 P=0.477N P=0.391



### Pituitary Gland (Pars Intermedia): Adenoma

Overall rate 0/46 (0%) 0/49 (0%) 4/47 (9%) 0/48 (0%)

Adjusted rate 0.0% 0.0% 9.4% 0.0%

Terminal rate 0/34 (0%) 0/39 (0%) 4/35 (11%) 0/41 (0%)

First incidence (days)

— — 731 (T)

— Poly

- 3 test P=0.551

— f P=0.064

—



### Skin (Subcutaneous Tissue) : Sarcoma

Overall rate 7/50 (14%) 2/50 (4%) 3/50 (6%) 1/50 (2%)

Adjusted rate 15.0% 4.2% 6.6% 2.1%

Terminal rate 1/36 (3%) 0/40 (0%) 2/37 (5%) 1/41 (2%)

First incidence (days) 562 505 610 731 (T)

Poly

- 3 test P=0.029N P=0.074N P=0.169N P=0.029N



### Skin (Subcutaneous Tissue) : Fibrous Histiocytoma, Fibrosarcoma, or Sarcoma

Overall rate 8 /50 (1 6 %) 4 /50 ( 8 %) 4 /50 ( 8 %) 1/50 (2%)

Adjusted rate 1 7.1 % 8.3 % 8.7 % 2.1%

Terminal rate 2 /36 ( 6 %) 1 /40 ( 3 %) 2/37 (5%) 1/41 (2%)

First incidence (days) 562 505 537 731 (T)

Poly

- 3 test P=0.0 14 N P=0. 16 4N P=0.1 84 N P=0.0 15 N

### 140 1- Bromopropane, NTP TR 564

T ABLE 

### D2

Statistical Analysis of Primary Neoplasms in Fem ale Mice in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

Uterus: Stromal Polyp

Overall rate 0/50 (0%) 2/50 (4%) 2/50 (4%) 3/50 (6%)

Adjusted rate 0.0% 4.2% 4.4% 6.4%

Terminal rate 0/36 (0%) 1/40 (3%) 1/37 (3%) 2/41 (5%)

First incidence (days)

— 547 589 719

Poly

- 3 test P=0.121 P=0.252 P=0.241 P=0.129



### All Organs: Hemangiosarcoma

Overall rate 4/50 (8%) 2/50 (4%) 2/50 (4%) 1/50 (2%)

Adjusted rate 9.0% 4.3% 4.4% 2.1%

Terminal rate 4/36 (11%) 1/40 (3%) 1/37 (3%) 1/41 (2%)

First incidence (days) 731 (T) 687 603 731 (T)

Poly

- 3 test P=0.131N P=0.312N P=0.328N P=0.162N



### All Organs: Hemangioma or Hemangiosarcoma

Overall rate 4/50 (8%) 3/50 (6%) 2/50 (4%) 1/50 (2%)

Adjusted rate 9.0% 6.4% 4.4% 2.1%

Terminal rate 4/36 (11%) 2/40 (5%) 1/37 (3%) 1/41 (2%)

First incidence (days) 731 (T) 687 603 731 (T)

Poly

- 3 test P=0.107N P=0.470N P=0.328N P=0.162N



### All Organs: Malignant Lymphoma

Overall rate 17/50 (34%) 9/50 (18%) 15/50 (30%) 11/50 (22%)

Adjusted rate 37.2% 19.0% 33.0% 23.2%

Terminal rate 13/36 (36%) 8/40 (20%) 13/37 (35%) 8/41 (20%)

First incidence (days) 591 583 631 649

Poly

- 3 test P=0.198N P=0.040N P=0.421N P=0.104N



### All Organs: Benign Neoplasms

Overall rate 22/50 (44%) 24/50 (48%) 30/50 (60%) 28/50 (56%)

Adjusted rate 47.2% 50.3% 64.0% 57.3%

Terminal rate 17/36 (47%) 21/40 (53%) 25/37 (68%) 21/41 (51%)

First incidence (days) 481 547 589 470

Poly

- 3 test P=0.150 P=0.463 P=0.072 P=0.217



### All Organs: Malignant Neoplasms

Overall rate 33/50 (66%) 28/50 (56%) 33/50 (66%) 22/50 (44%)

Adjusted rate 68.4% 57.0% 67.9% 45.2%

Terminal rate 22/36 (61%) 20/40 (50%) 23/37 (62%) 15/41 (37%)

First incidence (days) 540 505 345 568

Poly

- 3 test P=0.021N P=0.168N P=0.566N P=0.016N

### 1- Bromopropane, NTP TR 564 141

T ABLE 

### D2

Statistical Analysis of Primary Neoplasms in Fem ale Mice in the 2

- Year Inhalation Study of 1

- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

All Organs: Benign or Malignant Neoplasms

Overall rate 42/50 (84%) 37/50 (74%) 44/50 (88%) 34/50 (68%)

Adjusted rate 85.0% 74.4% 89.6% 68.8%

Terminal rate 29/36 (81%) 28/40 (70%) 33/37 (89%) 26/41 (63%)

First incidence (days) 481 505 345 470

Poly

- 3 test P=0.064N P=0.142N P=0.349 P=0.044N

(T) Terminal sacrifice

a Number of neoplasm

- bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal

medulla , liver, lung , ovary, and pituitary gland ; for other tissues, denominator is number of animals necropsied.

b Poly

- 3 estimated neoplasm incidence after adjustment for intercurrent mortality

c Observed incidence at terminal kill

d Beneath the chamber control incidence is the P value associated with the trend test. Beneath the exposed group incidence are the P values

corresponding to pairwise comparisons between the chamber controls and that expos ed group. The Poly

- 3 test accounts for the differential

mortality in animals that do not reach terminal sacrifice. A negative trend or a lower incidence in an expos ur e group is indicated by N

.

e Not applicable; no neoplasms in animal group

f Value of statistic cannot be computed

.

### 142 1- Bromopropane, NTP TR 564

T ABLE 

### D3

Historical Incidence of Alveolar/bronchiolar Neoplasms in Control Female B6C3F1 Mice a



### Study (Study Start) Incidence in Controls

Adenoma Carcinoma Adenoma or Carcinoma

Historical Incidence: Inhalation Studies

1

- Bromopropane (July 2003) 1/50 0/50 1 /50

Cumene (June 2001) 1/50 3 /50 4 /50

Diethylamine (August 2003) 2/50 3 /50 5 /50

Methyl isobutyl ketone ( June 2000) 4/50 0/50 4 /50

α

- Methylst y rene (July 2001) 1/50 1 /50 2 /50

Propargyl alcohol (September 2001) 3/50 2 /50 5 /50

Tetralin (June 2003) 6/50 0/50 6 /50

Total (%) 18 /350 ( 5 .1%) 9 /350 (2.6%) 27 /350 (7.7%)

Mean 

### ± standard deviation 5 .1% 

### ± 3.8 % 2.6 % 

### ± 2.8 % 7.7 % ± 3.6 %

Range 2%-1 2% 0% -6% 2%-1 2%



### Overall Historical Incidence: All Routes

Total (%) 7 3/1, 496 ( 4.9 %) 59 /1 ,496 ( 3.9 %) 128 /1, 496 ( 8.6 %)

Mean 

### ± standard deviation 4.9 % 

### ± 3.4 % 3.9 % 

### ± 3 .4% 8.6 % ± 3 .7%

Range 0%-1 2% 0% -1 2% 2%- 18 %

a Data as of April 29, 2009

### 1- Bromopropane, NTP TR 564 143

T ABLE 

### D4

Summary of the Incidence of Nonneoplastic Lesions in Fem ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane a



### Chamber Control 62.5 ppm 125 ppm 250 ppm

Disposition Summary

Animals initially in study 50 50 50 50

Early deaths

Moribund 10 7 8 8

Natural deaths 4 3 5

Survivors

Died last week of study 1 1

Terminal sacrifice 3 5 40 37 4 1

Animals examined microscopically 50 50 50 50



### Alimentary System

Esophagus (50) (50) (50) (50)

Gallbladder (43) (36) (36) (44)

Inflammation, c hronic a ctive 1 (3%)

Intestine l arge, c ecum (48) (49) (46) (50)

Hemorrhage 1 (2%)

Inflammation, s uppurative 1 (2%)

Intestine l arge, c olon (48) (49) (46) (50)

Necrosis 1 (2%)

Intestine l arge, r ectum (47) (49) (46) (50)

Artery, i nflammation, c hronic a ctive 1 (2%)

Intestine s mall, d uodenum (46) (49) (46) (50)

Intestine s mall, i leum (48) (49) (46) (50)

Epithelium, h yperplasia 1 (2%)

Intestine s mall, j ejunum (46) (49) (46) (50)

Epithelium, h yperplasia 1 (2%)

Liver (50) (50) (50) (50)

Angiectasis 1 (2%) 1 (2%) 2 (4%)

Basophilic f ocus 2 (4%) 3 (6%) 2 (4%) 3 (6%)

Clear c ell f ocus 2 (4%) 1 (2%) 2 (4%)

Eosinophilic f ocus 6 (12%) 7 (14%) 7 (14%) 5 (10%)

Fatty c hange 1 (2%) 1 (2%) 2 (4%)

Fatty c hange, f ocal 2 (4%)

Hematopoietic c ell p roliferation 1 (2%)

Infarct 1 (2%)

Inflammation, a cute 1 (2%)

Inflammation, c hronic 1 (2%) 1 (2%)

Malformation, l obular 1 (2%)

Mineralization 1 (2%)

Mixed c ell f ocus 2 (4%) 3 (6%) 3 (6%)

Necrosis 1 (2%) 1 (2%) 2 (4%) 1 (2%)

Tension l ipidosis 3 (6%) 10 (20%) 4 (8%) 1 (2%)

Thrombosis 1 (2%)

Vacuolization c ytoplasmic 1 (2%)

Bile d uct, h yperplasia 1 (2%)

Hepatocyte, m itotic a lteration 1 (2%)

Mesentery (11) (9) (11) (13)

Artery, i nflammation 1 (9%)

Fat, h emorrhage 1 (11%) 1 (9%)

Fat, n ecrosis 9 (82%) 9 (100%) 9 (82%) 1 2 (92%)

Pancreas (50) (49) (50) (50)

Atrophy 7 (14%) 4 (8%) 5 (10%) 2 (4%)

Basophilic f ocus 1 (2%) 2 (4%)

Inflammation, c hronic a ctive 1 (2%) 1 (2%)

Salivary g lands (50) (50) (50) (50)

a Number of animals examined microscopically at the site and the number of animals with lesion

### 144 1- Bromopropane, NTP TR 564

T ABLE 

### D4

Summary of the Incidence of Nonneoplastic Lesions in Fem ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

Alimentary System (continued)

Stomach, f orestomach (50) (50) (49) (50)

Hyperplasia, s quamous 3 (6%) 2 (4%) 2 (4%) 1 (2%)

Inflammation 1 (2%)

Ulcer 2 (4%) 2 (4%) 2 (4%)

Artery, i nflammation, c hronic a ctive 1 (2%)

Stomach, g landular (50) (49) (48) (50)

Infiltration c ellular, m ixed c ell 1 (2%)

Infiltration c ellular, m ononuclear c ell 1 (2%)

Mineralization 1 (2%) 1 (2%)

Necrosis 1 (2%) 1 (2%)

Artery, i nflammation, c hronic a ctive 1 (2%)

Tongue (0) (0) (1) (0)

Tooth (0) (1) (0) (0)

Dysplasia 1 (100%)



### Cardiovascular System

Heart (50) (50) (50) (50)

Cardiomyopathy 6 (12%) 12 (24%) 8 (16%) 8 (16%)

Inflammation, s uppurative 1 (2%)

Mineralization 3 (6%) 2 (4%)

Necrosis 1 (2%)

Thrombosis 2 (4%)

Artery, i nflammation, c hronic a ctive 1 (2%) 1 (2%)



### Endocrine System

Adrenal c ortex (50) (50) (50) (50)

Hyperplasia 3 (6%) 10 (20%) 4 (8%) 6 (12%)

Hypertrophy 4 (8%) 3 (6%) 1 (2%) 9 (18%)

Inflammation, s uppurative 1 (2%)

Subcapsular, h yperplasia 1 (2%) 1 (2%)

Adrenal m edulla (50) (50) (49) (50)

Hyperplasia 5 (10%) 1 (2%) 4 (8%)

Islets, p ancreatic (50) (49) (49) (49)

Hyperplasia 2 (4%) 1 (2%)

Pituitary g land (46) (49) (47) (48)

Pars d istalis, a ngiectasis 1 (2%) 1 (2%) 2 (4%)

Pars d istalis, h yperplasia 15 (33%) 14 (29%) 11 (23%) 15 (31%)

Pars i ntermedia, h yperplasia 1 (2%) 2 (4%)

Pars i ntermedia, h ypertrophy 1 (2%) 1 (2%)

Thyroid g land (49) (50) (46) (50)

Follicular c ell, h yperplasia 1 (2%) 1 (2%) 1 (2%) 1 (2%)



### General Body System

Peritoneum (0) (0) (0) (1)

### 1- Bromopropane, NTP TR 564 145

T ABLE 

### D4

Summary of the Incidence of Nonneoplastic Lesions in Fem ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

G enital S ystem

Ovary (50) (50) (50) (49)

Angiectasis 1 (2%) 2 (4%) 2 (4%)

Congestion, c hronic 1 (2%)

Cyst 12 (24%) 8 (16%) 12 (24%) 2 (4%)

Hyperplasia, t ubular 1 (2%)

Thrombosis 2 (4%) 2 (4%) 1 (2%)

Uterus (50) (50) (50) (50)

Angiectasis 2 (4%) 3 (6%) 6 (12%) 3 (6%)

Fibrosis 1 (2%)

Inflammation, s uppurative 1 (2%)

Inflammation, a cute 1 (2%)

Thrombosis 1 (2%) 1 (2%) 2 (4%) 1 (2%)

Endometrium, h yperplasia, c ystic 39 (78%) 43 (86%) 43 (86%) 41 (82%)



### Hematopoietic System

Bone m arrow (50) (50) (49) (50)

Angiectasis 2 (4%)

Atrophy 1 (2%)

Lymph n ode (6) (5) (3) (8)

Iliac, a ngiectasis 2 (33%) 1 (20%)

Iliac, h yperplasia, l ymphoid 1 (17%)

Lumbar, a ngiectasis 1 (20%) 1 (33%) 1 (13%)

Renal, a ngiectasis 1 (13%)

Renal, h yperplasia, l ymphoid 1 (17%) 1 (20%)

Lymph n ode, b ronchial (37) (32) (39) (34)

Lymph n ode, m andibular (45) (30) (30) (37)

Hyperplasia, l ymphoid 1 (3%)

Lymph n ode, m ediastinal (45) (39) (41) (37)

Hyperplasia, l ymphoid 1 (2%)

Lymph n ode, m esenteric (48) (47) (47) (49)

Angiectasis 2 (4%) 2 (4%) 1 (2%)

Hyperplasia, l ymphoid 1 (2%)

Spleen (50) (49) (50) (50)

Atrophy 4 (8%) 1 (2%) 2 (4%) 5 (10%)

Congestion 1 (2%) 2 (4%)

Hematopoietic c ell p roliferation 13 (26%) 10 (20%) 13 (26%) 16 (32%)

Hemorrhage 1 (2%)

Hyperplasia, h istiocytic 2 (4%)

Hyperplasia, l ymphoid 7 (14%) 7 (14%) 5 (10%) 3 (6%)

Hyperplasia, p lasma c ell 1 (2%)

Necrosis, l ymphoid 1 (2%) 1 (2%)

Pigmentation, h emosiderin 1 (2%) 2 (4%)

Thymus (49) (45) (45) (45)

Atrophy 14 (29%) 13 (29%) 17 (38%) 16 (36%)

Cyst 5 (10%) 7 (16%) 6 (13%) 9 (20%)

Necrosis, l ymphoid 2 (4%) 4 (9%)

Medulla, h yperplasia, l ymphoid 25 (51%) 29 (64%) 30 (67%) 26 (58%)



### Integumentary System

Mammary g land (50) (50) (50) (50)

Hyperplasia 1 (2%) 1 (2%) 1 (2%)

Skin (50) (50) (50) (50)

Cyst 1 (2%)

Inflammation, c hronic a ctive 4 (8%) 2 (4%) 1 (2%)

### 146 1- Bromopropane, NTP TR 564

T ABLE 

### D4

Summary of the Incidence of Nonneoplastic Lesions in Fem ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

M usculoskeletal S ystem

Bone (50) (50) (50) (50)

Cyst 1 (2%)

Skeletal m uscle (0) (0) (0) (1)



### N ervous S ystem

Brain (50) (50) (50) (50)

Necrosis 1 (2%) 1 (2%)

Artery, i nflammation 1 (2%)

Peripheral n erve (1) (1) (1) (1)

Degeneration 1 (100%)

Spinal c ord (1) (1) (1) (1)

Inflammation, c hronic a ctive 1 (100%)



### R espiratory S ystem

Larynx (50) (50) (50) (50)

Vacuolization c ytoplasmic 3 (6%) 2 (4%) 2 (4%)

Artery, i nflammation, c hronic a ctive 1 (2%)

Lung (50) (50) (50) (50)

Inflammation, c hronic a ctive 1 (2%)

Alveolar e pithelium, h yperplasia 1 (2%) 1 (2%) 3 (6%)

Alveolus, i nfiltration c ellular, h istiocyte 4 (8%) 1 (2%)

Bronchiole, h yperplasia 1 (2%) 1 (2%)

Bronchiole, n ecrosis 1 (2%)

Bronchiole, r egeneration 45 (90%) 43 (86%) 49 (98%)

Bronchiole, v acuolization c ytoplasmic 3 (6%) 4 (8%) 3 (6%)

Nose (50) (50) (50) (50)

Inflammation, s uppurative 1 (2%)

Olfactory e pithelium, a trophy 6 (12%)

Olfactory e pithelium, m etaplasia,

r espiratory 4 (8%) 5 (10%) 14 (28%)

Respiratory e pithelium, h yperplasia 11 (22%) 25 (50%) 28 (56%) 27 (54%)

Respiratory e pithelium, v acuolization

cytoplasmic 3 (6%) 5 (10%) 8 (16%)

Pleura (0) (0) (1) (0)

Trachea (50) (49) (50) (50)

Vacuolization c ytoplasmic 8 (16%) 7 (14%) 4 (8%)



### Special Senses System

Eye (47) (49) (45) (50)

Cataract 3 (6%) 1 (2%) 2 (4%)

Cornea, h yperplasia, s quamous 1 (2%)

Cornea, i nflammation, c hronic a ctive 1 (2%) 2 (4%) 1 (2%)

Cornea, m ineralization 1 (2%) 2 (4%) 1 (2%)

Harderian g land (49) (49) (48) (50)

Hyperplasia 3 (6%) 1 (2%) 2 (4%) 6 (12%)

Inflammation, c hronic a ctive 1 (2%)

Zymbal ’ s g land (0) (1) (0) (0)

### 1- Bromopropane, NTP TR 564 147

T ABLE 

### D4

Summary of the Incidence of Nonneoplastic Lesions in Fem ale Mice

in the 2

- Year Inhalation Study of 1- Bromopropane

Chamber Control 62.5 ppm 125 ppm 250 ppm

U rinary S ystem

Kidney (50) (49) (49) (50)

Amyloid d eposition 1 (2%)

Cyst 1 (2%)

Inflammation, s uppurative 1 (2%)

Inflammation, c hronic a ctive 1 (2%)

Metaplasia, o sseous 3 (6%) 4 (8%) 2 (4%)

Nephropathy 33 (66%) 37 (76%) 40 (82%) 39 (78%)

Renal t ubule, h yperplasia 1 (2%)

Transitional e pithelium, h yperplasia 1 (2%)

Urinary b ladder (50) (49) (48) (50)

Inflammation, c hronic a ctive 1 (2%)

Artery, i nflammation, c hronic a ctive 2 (4%)

### 148 1- Bromopropane, NTP TR 564

### 149



# APPENDIX E

GENETIC TOXICOLOGY

B ACTERIAL 

### M UTAGENICITY 

### T EST 

### P ROTOCOL



### ....................................................................................................... 150

M OUSE 

### P ERIPHERAL 

### B LOOD 

### M ICRONUCLEUS 

### T EST 

### P ROTOCOL



### ........................................................................... 150

E VALUATION 

### P ROTOCOL



### ................................................................................................ ......................................... 151

R ESULTS



### ................................................................................................ .................................................................... 151

T ABLE 

### E1 Mutagenicity of 1- Bromopropane in Bacterial Tester Strains

....................................................... 152

T ABLE 

### E2 Frequency of Micronuclei in Peripheral Blood Erythrocytes of Mice

Following Treatment with 1- Bromopropane by Inhalation for 3 Months

.................................... 154

### 150 1- Bromopropane, NTP TR 564



# GENETIC TOXICOLOGY

B ACTERIAL 

# M UTAGENICITY 

# T EST 

# P ROTOCOL

Testing at Bio R eliance Corporation was performed as reported by Zeiger et al. ( 1992 ). 1- Bromopropane was sent to

the laboratory as a coded aliquot. In the tests conducted at Bio R eliance Corporation , i t was incubated with the

Salmonella typhimurium tester strains TA97, TA98, TA100, and TA1535 either in buffer or S9 mix (metabolic

activation enzymes and cofactors from Aroclor 1254

- induced male Sprague

- Dawley rat or Syrian hamster liver) for

20 minutes at 37° C. Top agar supplemented with L



### - histidine and d

- biotin was added, and the contents of t he tubes

were mixed and poured onto the surfaces of minimal glucose agar plates. Histidine

- independent mutant colonies

arising on these plates were counted following incubation for 2 days at 37° C. The slightly modified protocol used

at SITEK Research La boratories tested the same lot (1581313004) of 1

- bromopropane used in the 2

- year studies

, used only 10% rat liver S9 for exogenous metabolic activation

, and employed Escherichia coli strain

WP2 uvrA / pKM101 as a bacterial tester strain in addition to S. typ himurium strains TA98 and TA100. Incubati on of

bacterial strains with 1

- bromopropane and subsequent plating were carried out as described above for the traditional

protocol.

Each trial consisted of triplicate plates of concurrent positive and negative controls and five doses of

1- bromopropane . The high dose was limited by toxicity in some trials and by the limit dose of 10,000 μg/plate in

those trials where only slight toxici ty was observed. All trials were repeated

, and those that were conducted with S9

activation enzymes were repeated using the same or higher concentrations of S9.

In this assay, a positive response is defined as a reproducible, dose

- related increase in histidine

- independent

(revertant) colonies in any one strain/activation combination. An equivocal response is defined as an increase in

revertants that is not dose related, is not reproducible, or is not of sufficient magnitude to support a determination of

mutagenicity. A negative response is obtained when no increase in revertant colonies is observed following

chemical treatment. There is no minimum percentage or fold increase required for a chemical to be judged positive

or weakly positive, although p ositive calls are typically reserved for increases in mutant colonies that are at least

twofold over background

.



# M OUSE 

# P ERIPHERAL 

# B LOOD 

# M ICRONUCLEUS 

# T EST 

# P ROTOCOL

A detailed discussion of this assay is presented by MacGregor et al. (1990). At the end o f the 3- month toxicity

study , peripheral blood samples were obtained from male and female mice. Smears were immediately prepared and

fixed in absolute methanol. The methanol

- fixed slides were stained with a cridine orange and coded. Slides were

scanned to determine the frequency of micronuclei in 2 ,000 normochromatic erythrocytes (NCEs ; immature

erythrocytes ) per animal. In addition, the percentage of polychromatic erythrocytes ( PCEs or reticulocytes) in a

population of 1,000 erythrocytes was determined as a measure of bone marrow toxicity.

The results were tabulated as the mean of the pooled results from all animals within a n exposure group plus or

minus the standard error of the mean. The frequency of micronucleated cells among NCEs was analyzed by a

statistical software package that tested for increasing trend over exposure groups with a one

- tailed

Cochran

- Armitage trend test, followed by pairwise comparisons between each expos ed group and the chamber

control group. In the presence of excess binomial variation, as detected by a binomial dispersion test, the binomial

variance of the Cochran

- Armitage test was adjusted upward in proportion to the excess variation. In the

micronucleus test, an individual trial is considered positive if the trend test P value is less than or equal to 0.025 or if

the P value for any single exposed group is less than or equal to 0.025 divided by the number of expos ed groups. A

final call of positive for micronucleus induction is preferably based on reprod ucibly positive trials (as noted above).

Results of the 3- month stud y were accepted without repeat tests, because additional test data could not be obtained.

Ultimately, the final call is determined by the scientific staff after considering the results o f statistical analyses, the

reproducibility of any effects observed, and the magnitudes of those effects.

### 1- Bromopropane, NTP TR 564 151



# E VALUATION 

# P ROTOCOL

These are the basic guidelines for arriving at an overall assay result for assays performed by the National

Toxicology Program . Statistical as well as biological factors are considered. For an individual assay, the statistical

procedures for data analysis have been described in the preceding protocols. There have been instances, however, in

which multiple aliquots of a chemica l were tested in the same assay, and different results were obtained among

aliquots and/or among laboratories. Results from more than one aliquot or from more than one laboratory are not

simply combined into an overall result. Rather, all the data are cr itically evaluated, particularly with regard to

pertinent protocol variations, in determining the weight of evidence for an overall conclusion of chemical activity in

an assay. In addition to multiple aliquots, the in vitro assays have another variable that must be considered in

arriving at an overall test result. In vitro assays are conducted with and without exogenous metabolic activation.

Results obtained in the absence of activation are not combined with results obtained in the presence of activation ;

each testing condition is evaluated separately. The summary table in the Abstract of this Technical Report presents

a result that represents a scientific judgment of the overall evidence for activity of the chemical in an assay

.



# R ESULTS

1- Bromopropane (doses up to 10,000 μ g/plate) was not mutagenic in either of two independent bacterial

mutagenicity assays, with or without exogenous metabolic activation (Table E1). Bacterial strains tested included

S. typhimurium strains TA97, TA98, TA100, and TA1535, as well as E. coli strain WP2 uvrA / pKM101. In addition

to the negative results in the two bacterial tests, no increases in the frequencies of micronucleated NCE s were seen

in peripheral blood of male or female B6C3F1 mice exposed for 3 months to 6 2.5 to 500 ppm 1

- bromopropane via

inhalation (Table E2). The percentage of reticulocytes ( PCEs ) in the peripheral blood of male and female mice was

unaltered by 1

- bromopropane exposure, suggesting a lack of chemical

- associated bone marrow toxicity.

### 152 1- Bromopropane, NTP TR 564

T A BLE 

### E1

Mutagenicity of 1- Bromopropane in Bacterial Tester Strains a



### Revertants/Plate b



### Dose −S9 +hamster S9 +rat S9

Strain (μg/Plate) Trial 1 Trial 2 10% 30% 10% 30%

Study performed at BioReliance Corporation

TA100 0 150 ± 4 121 ± 16 104 ± 2 103 ± 9 124 ± 2 140 ± 8

33 115 ± 6 118 ± 6 121 ± 7

1 0 0 144 ± 1 0 1 24 ± 4 1 09 ± 1 2 1 0 4 ± 8 1 23 ± 2 13 3 ± 5

3 3 3 1 39 ± 7 1 27 ± 7 92 ± 8 108 ± 6 1 28 ± 10 1 1 7 ± 12

1

, 0 0 0 149 ± 7 1 28 ± 6 111 ± 7 98 ± 6 14 4 ± 5 1 03 ± 7

3, 333 117 ± 2 c 107 ± 11 c 98 ± 19 c 118 ± 6 c 10 7 ± 7 c 123 ± 7 c

1 0, 000 Toxic Toxic Toxic

Trial summary Negative Negative Negative Negative Negative Negative

Positive control d 728 ± 27 279 ± 14 6 61 ± 22 615 ± 14 5 23 ± 32 636 ± 27



### TA1535 0 16 ± 2 10 ± 2 10 ± 2 12 ± 2 12 ± 2 1 4 ± 3

33 9 ± 1 9 ± 2 9 ± 2

10 0 19 ± 2 11 ± 0 1 2 ± 2 18 ± 2 8 ± 1 e 14 ± 2

33 3 1 4 ± 3 10 ± 1 1 1 ± 2 1 5 ± 4 8 ± 1 1 8 ± 3

1

, 00 0 14 ± 1 10 ± 1 12 ± 1 17 ± 1 1 3 ± 2 1 5 ± 0

3, 333 6 ± 3 f 9 ± 1 c 8 ± 1 c 10 ± 1 c 8 ± 2 c 8 ± 0 c

1 0

, 000 2 ± 1 c 6 ± 1 c 4 ± 1 c

Trial summary Negative Negative Negative Negative Negative Negative

Positive control 226 ± 10 19 9 ± 19 6 0 ± 6 13 7 ± 9 12 7 ± 13 80 ± 8



### TA97 0 134 ± 27 1 05 ± 12 1 5 6 ± 6 12 5 ± 9 1 61 ± 7 1 68 ± 13

3 3 1 30 ± 12 16 3 ± 13 1 56 ± 9

10 0 10 8 ± 9 1 21 ± 3 11 4 ± 17 1 41 ± 11 1 41 ± 9 1 70 ± 9

33 3 98 ± 5 1 32 ± 6 1 24 ± 4 1 33 ± 12 1 49 ± 1 4 1 68 ± 5

1

, 00 0 1 17 ± 4 1 2 5 ± 6 1 43 ± 7 166 ± 3 e 147 ± 8 18 0 ± 13

3, 333 86 ± 3 c 1 17 ± 7 c 1 13 ± 9 c 91 ± 16 c 1 22 ± 10 c 1 21 ± 13 c

1 0,000 40 ± 22 f 53 ± 3 c 70 ± 15 c

Trial summary Negative Negative Negative Negative Negative Negative

Positive control 354 ± 17 2 11 ± 4 988 ± 30 1,063 ± 1 38 1,084 ± 3 2 418 ± 16



### TA98 0 23 ± 3 14 ± 1 1 6 ± 1 29 ± 1 25 ± 2 31 ± 4

3 3 1 8 ± 2 21 ± 3 24 ± 3

10 0 2 3 ± 2 15 ± 2 2 4 ± 3 2 3 ± 2 2 2 ± 2 27 ± 3

3 3 3 1 7 ± 2 16 ± 1 14 ± 1 1 7 ± 2 21 ± 5 2 3 ± 2

1 ,0 00 1 9 ± 2 1 5 ± 1 20 ± 2 2 2 ± 1 20 ± 2 2 1 ± 1

3 ,3 33 1 9 ± 1 1 7 ± 2 c 1 2 ± 3 c 1 9 ± 2 c 1 5 ± 1 c 15 ± 2 c

10, 000 Toxic 1 1 ± 2 c 13 ± 0 c

Trial summary Negative Negative Negative Negative Negative Negative

Positive control 340 ± 17 166 ± 3 875 ± 2 4 5 78 ± 17 302 ± 22 824 ± 2 8

### 1- Bromopropane, NTP TR 564 153

T ABLE 

### E1

Mutagenicity of 1- Bromopropane in Bacterial Tester Strains

Revertants/Plate

Dose

− S9 +10% rat S9

Strain (μg/Plate) Trial 1 Trial 2 Trial 1 Trial 2

Study Performed at SITEK Research Laboratories (lot 1581313004 used in the 2

- year studies)

TA100 0 6 2 ± 2 67 ± 4 76 ± 3 65 ± 2

5 00 57 ± 6 57 ± 3

1 ,00 0 62 ± 4

1,50 0 6 4 ± 3 53 ± 4

2,5 00 50 ± 2 35 ± 1 84 ± 2 67 ± 3

3,500 38 ± 6 16 ± 1 c 75 ± 3 63 ± 6

5 ,00 0 11 ± 2 c 6 6 ± 3 66 ± 4

7

, 5 00 62 ± 5 60 ± 4

1 0

, 0 00 59 ± 7 53 ± 7

Trial summary Negative Negative Negative Negative

Positive control 488 ± 27 635 ± 35 878 ± 19 982 ± 37



### TA98 0 18 ± 1 2 2 ± 3 2 9 ± 3 24 ± 2

5 00 1 6 ± 0 16 ± 1

1 ,00 0 19 ± 2

1,50 0 1 7 ± 3 18 ± 1

2,5 00 1 8 ± 2 14 ± 1 2 4 ± 0 2 3 ± 1

3,500 8 ± 2 c 5 ± 1 c 21 ± 4 2 1 ± 0

5 ,00 0 2 ± 1 c 2 6 ± 4 2 6 ± 1

7,5 00 23 ± 3 2 0 ± 1

1 0,000 6 ± 1 c 8 ± 2 c

Trial summary Negative Negative Negative Negative

Positive control 693 ± 2 6 693 ± 26 1

, 163 ± 77 742 ± 37



### Escherichia coli WP2 uvrA /pKM101

0 177 ± 3 1 63 ± 7 185 ± 8 1 90 ± 23

5 00 197 ± 25 190 ± 15

1,50 0 211 ± 20 1 94 ± 6

2,5 00 153 ± 7 1 39 ± 12 226 ± 17 214 ± 14

3,500 1 48 ± 10 1 2 4 ± 3 203 ± 1 4 20 1 ± 8

5 ,00 0 88 ± 3 c 7 6 ± 1 7 c 1 9 2 ± 6 1 95 ± 10

7,5 00 1 71 ± 6 16 4 ± 25

1 0,0 00 149 ± 7 142 ± 4

Trial summary Negative Negative Negative Negative

Positive control 1,883 ± 20 1, 955 ± 18 993 ± 79 1,1 64 ± 25

a The detailed protocol for the BioReliance Corporation study is presented by Zeiger et al. (1992); the study performed at SITEK Research

Laboratories used a slight modification of those procedures. 0 μ g/plate was the solvent control.

b Revertants are presented as mean ± standard error from three plates.

c Slight toxicity

d The positive controls in the absence of metabolic activation were sodium azide ( TA100 and TA1535 ), 9

- aminoacridine ( TA97 ),

4- nitro

- o- phenylenediamine (TA98), and methyl methanesulfonate ( E. coli ). The positive control for metabolic activation with all strains was

2- aminoanthracene.

e Contamination

f Slight toxicity and precipitate

### 154 1- Bromopropane, NTP TR 564

T ABLE 

### E2

Frequency of Micronuclei in Peripheral Blood Erythrocytes of Mice

Following Treatment with 1- Bromopropane by Inhalation for 3 Months a



### Compound Exposure

Concentration

(ppm) Number of Mice

with Erythrocytes

Scored Micronucleated NCEs/

1,000 NCEs b 

### P value c 

### PCEs (%) b



### Male

Air d 0 5 2.00 ± 0. 61 2. 52 ± 0. 30

1

- Bromopropane 62.5 5 3.10 ± 0. 81 0. 0615 2.1 6 ± 0. 31

125 5 2.70 ± 0. 64 0. 1533 2. 96 ± 0 .1 1

250 5 1.30 ± 0. 41 0. 8887 2.80 ± 0. 18

500 5 2.30 ± 0. 46 0.32 35 2.8 0 ± 0.4 4

P= 0.757 e



### Female

Air 0 5 1.80 ± 0. 25 2.24 ± 0. 52

1

- Bromopropane 62.5 5 1.70 ± 0. 25 0. 5672 2 .94 ± 0. 23

125 5 1.60 ± 0 .19 0. 6343 2. 74 ± 0. 52

250 5 1.40 ± 0. 33 0. 7604 2. 42 ± 0.2 7

5 0 0 5 1.80 ± 0. 20 0. 5000 2. 48 ± 0. 29

P=0. 500

a S tudy was performed at ILS, Inc. The detailed protocol is presented by MacGregor et al. (1990).

PCE=polychromatic erythrocyte; NCE=normochromatic erythrocyte.

b Mean ± standard error

c Pairwise comparison with the chamber control group; significant at P ≤ 0 .00 6

d Chamber control

e Significance of micronucleated NCEs/1,000 NCEs tested by the one

- tailed trend test ; significant at P ≤0 .025

### 155



# APPENDIX F

CLINICAL PATHOLOGY R ESULTS

T ABLE 

### F1 Hematology and Clinical Chemistry Data for Rats in the 3- Month Inhalation Study

of 1- Bromopropane

............................................................................................................................ 156

T ABLE 

### F2 Hematology Data for Mice in the 3- Month Inhalation Study of 1- Bromopropane

...................... 161

### 156 1- Bromopropane, NTP TR 564

T ABLE 

### F1

Hematology and Clinical Chemistry Data for Rats in the 3- Month Inhalation Study of 1

- Bromopropane a



### Chamber

Control 62.5 ppm 125 ppm 250 ppm 500 ppm 1,0 0 0 ppm

Male

Hematology

n

Day 3 10 10 10 9 10 10

Day 2 3 10 10 10 10 10 10

Week 14 10 10 10 10 10 10

Hematocrit (%)

Day 3 45.3 ± 0. 5 44.2 ± 0. 4 44.7 ± 0. 4 44.9 ± 0. 5 45.0 ± 0. 6 47.0 ± 0. 6

Day 2 3 47.2 ± 0. 5 47 .2 ± 0.4 47.9 ± 0.4 47.4 ± 0. 4 47.2 ± 0. 5 47.9 ± 0.5

Week 14 48.2 ± 0. 2 48.4 ± 0. 4 48.0 ± 0. 3 48.5 ± 0. 3 47.9 ± 0.3 49.1 ± 0. 5

Packed cell volume ( mL/dL )

Day 3 44.3 ± 0. 6 43.8 ± 0. 4 43.8 ± 0. 4 44.3 ± 0. 4 44.5 ± 0. 5 46.5 ± 0. 5**

Day 2 3 46.0 ± 0. 6 46.0 ± 0. 5 46.8 ± 0.4 46.4 ± 0. 5 46.1 ± 0. 4 47.1 ± 0. 4

Week 14 47.8 ± 0. 3 47.8 ± 0. 3 47.7 ± 0. 4 47.8 ± 0. 3 47.6 ± 0. 4 48.4 ± 0. 5

Hemoglobin (g/dL)

Day 3 13.5 ± 0.2 13.5 ± 0.1 13.6 ± 0.2 13.8 ± 0.2 13.7 ± 0. 1 14.3 ± 0. 2 **

Day 2 3 14.8 ± 0. 2 14.7 ± 0. 2 14.9 ± 0. 1 14.8 ± 0. 2 14.7 ± 0. 1 14. 9 ± 0. 1

Week 14 15.4 ± 0. 1 15.5 ± 0.1 15.5 ± 0.1 15.6 ± 0.1 15.5 ± 0.1 15.8 ± 0. 2

Erythrocytes (10 6 /μL)

Day 3 7.12 ± 0. 10 7.07 ± 0.0 8 7.14 ± 0.0 9 7.30 ± 0. 08 7.34 ± 0.0 9 7.71 ± 0. 1 1 **

Day 2 3 7.80 ± 0. 10 7.74 ± 0.0 9 7.84 ± 0. 10 7.78 ± 0. 07 7.67 ± 0. 09 7.84 ± 0. 05

Week 14 9.07 ± 0. 06 9.02 ± 0. 05 9.01 ± 0.05 9.06 ± 0.0 4 8.98 ± 0.0 8 9.15 ± 0.0 9

Reticulocytes (10 3 /μL)

Day 3 360.8 ± 28.8 356.4 ± 32.6 381.7 ± 25.1 350.3 ± 30.4 362.4 ± 33.7 346.3 ± 17.0

Day 2 3 209.2 ± 19.6 187.6 ± 16.7 233.4 ± 19.1 240.4 ± 15.2 222.6 ± 10.8 222.0 ± 11.8

Week 14 106.1 ± 14.6 131.2 ± 12.4 137.4 ± 12.4 127.6 ± 15.7 97.8 ± 12.6 119.3 ± 10.7

Nucleated erythrocytes /100 leukocytes

Day 3 0. 70 ± 0. 15 1.10 ± 0. 28 0. 90 ± 0. 35 0. 78 ± 0 .28 0. 20 ± 0. 13 0. 10 ± 0.1 0*

Day 2 3 0. 20 ± 0. 13 0.00 ± 0.00 0. 1 0 ± 0. 1 0 0.00 ± 0.00 0. 3 0 ± 0. 15 0.00 ± 0.00

Week 14 0.0 0 ± 0.0 0 0. 3 0 ± 0. 15 0.1 0 ± 0.1 0 0. 30 ± 0. 2 1 0. 5 0 ± 0. 22 0. 3 0 ± 0. 21

Mean cell volume (fL)

Day 3 62 .2 ± 0. 2 61.9 ± 0. 4 61.4 ± 0. 4 60.7 ± 0. 3** 60.6 ± 0.4 ** 60.3 ± 0. 4* *

Day 2 3 59.0 ± 0.3 59.4 ± 0. 3 59.8 ± 0. 4 59.6 ± 0. 2 60.2 ± 0. 4* 60.0 ± 0. 3

Week 14 52.7 ± 0.2 53.0 ± 0. 2 52.9 ± 0.2 52.8 ± 0. 2 53.0 ± 0. 3 52.9 ± 0.2

Mean cell hemoglobin (pg)

Day 3 18.9 ± 0.1 19. 0 ± 0.1 19.0 ± 0. 1 1 8.9 ± 0. 1 1 8

. 7 ± 0.1 18. 6 ± 0.1 *

Day 2 3 18.9 ± 0. 1 19. 0 ± 0.1 19. 0 ± 0.1 19. 0 ± 0.1 19. 2 ± 0.1 19.0 ± 0.1

Week 14 17.1 ± 0. 0 17.2 ± 0.1 17.1 ± 0. 1 17.2 ± 0.1 17.3 ± 0. 1 17. 2 ± 0. 1

Mean cell hemoglobin concentration (g/dL)

Day 3 30.4 ± 0.1 30.7 ± 0. 2 30.8 ± 0. 2 31.1 ± 0.2 * 30.8 ± 0. 2 30.8 ± 0. 2

Day 2 3 32.1 ± 0.1 32.0 ± 0. 1 31.8 ± 0.1 31.8 ± 0.2 31.9 ± 0. 1 31.7 ± 0.1

Week 14 32.4 ± 0.1 32.5 ± 0.1 32.4 ± 0. 1 32.6 ± 0. 2 32.6 ± 0. 1 32. 5 ± 0. 2

Platelets (10 3 /μL)

Day 3 905.9 ± 33.7 875.4 ± 16.8 888.2 ± 16.3 799.0 ± 30.6** 759.8 ± 29.9** 820.9 ± 33.4* *

Day 2 3 754.5 ± 18.1 798.8 ± 14.0 820.2 ± 16.4* 802.4 ± 20.1 * 843.3 ± 14.8** 845.8 ± 15.6**

Week 14 678.6 ± 10.6 674.2 ± 8.3 653.8 ± 14.4 661.8 ± 13.9 647.8 ± 16.1 651.7 ± 6.4

Leukocytes (10 3 /μL)

Day 3 10.39 ± 0. 50 9.77 ± 0. 71 10.77 ± 0. 4 7 10.95 ± 0. 56 8. 97 ± 0.5 3 7.79 ± 0.4 3**

Day 2 3 7.90 ± 0. 53 8.40 ± 0. 40 8.13 ± 0. 57 7.73 ± 0. 32 8.24 ± 0. 44 7.68 ± 0. 44

Week 14 7.25 ± 0. 17 6.78 ± 0. 22 7.48 ± 0. 41 8.09 ± 0. 37 8.07 ± 0. 34 8.56 ± 0. 33*

Segmented neutrophils (10 3 /μL)

Day 3 1.22 ± 0. 06 1.19 ± 0. 06 1.34 ± 0.0 6 1.34 ± 0.0 9 1.10 ± 0.09 0.92 ± 0.0 5**

Day 2 3 1.16 ± 0. 03 1.25 ± 0.0 4 1.15 ± 0. 06 1.18 ± 0. 04 1.07 ± 0. 05 0. 93 ± 0. 03**

Week 14 1.19 ± 0. 09 1.18 ± 0.0 9 1.34 ± 0.1 1 1.32 ± 0. 07 1.40 ± 0. 08 1.37 ± 0.1 1

### 1- Bromopropane, NTP TR 564 157

T ABLE 

### F1

Hematology and Clinical Chemistry Data for Rats in the 3- Month Inhalation Study of 1

- Bromopropane

Chamber

Control 62.5 ppm 1 25 ppm 250 ppm 500 ppm 1, 0 00 ppm

Male (continued)

Hematology (continued)

n

Day 3 10 10 10 9 10 10

Day 2 3 10 10 10 10 10 10

Week 14 10 10 10 10 10 10

Bands (10 3 /μL)

Day 3 0.0 0 ± 0.0 0 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00

Day 2 3 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00

Week 14 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00

Lymphocytes (10 3 /μL)

Day 3 8.93 ± 0. 46 8.39 ± 0. 66 9.19 ± 0. 43 9.38 ± 0.4 9 7.65 ± 0.4 5 6.72 ± 0. 39**

Day 2 3 6.62 ± 0. 50 7.01 ± 0. 36 6.86 ± 0. 51 6.42 ± 0. 2 9 7.03 ± 0. 39 6.63 ± 0. 43

Week 14 5.79 ± 0. 17 5.35 ± 0. 16 5.73 ± 0. 35 6.49 ± 0. 34 6.33 ± 0. 34 6.91 ± 0. 27*

Monocytes (10 3 /μL)

Day 3 0.1 4 ± 0.0 2 0. 09 ± 0.01 0.1 4 ± 0.0 2 0. 09 ± 0.0 2 0. 14 ± 0.0 3 0. 08 ± 0.0 2*

Day 2 3 0. 0 4 ± 0.0 1 0. 06 ± 0.0 1 0. 04 ± 0.0 1 0. 05 ± 0.0 1 0.0 5 ± 0.0 1 0. 05 ± 0.0 1

Week 14 0.1 7 ± 0.0 5 0.1 6 ± 0.0 4 0. 27 ± 0.0 8 0. 18 ± 0.0 5 0. 22 ± 0.0 7 0. 2 1 ± 0.0 3

Basophils (10 3 /μL)

Day 3 0.00 7 ± 0.00 2 0.00 4 ± 0.00 2 0.00 6 ± 0.00 2 0.00 7 ± 0.00 2 0.00 3 ± 0.00 2 0.00 5 ± 0.00 2

Day 2 3 0.00 7 ± 0.00 2 0.00 5 ± 0.00 2 0.00 8 ± 0.00 2 0.00 6 ± 0.00 2 0.00 6 ± 0.00 2 0.00 7 ± 0.00 2

Week 14 0.00 2 ± 0.00 1 0.00 2 ± 0.00 2 0.0 1 0 ± 0.00 4 0.00 4 ± 0.00 2 0.00 3 ± 0.00 2 0.00 9 ± 0.00 9

Eosinophils (10 3 /μL)

Day 3 0.0 9 ± 0.01 0.10 ± 0.0 1 0.10 ± 0.01 0.13 ± 0.0 2 0.0 9 ± 0.01 0.0 6 ± 0.0 1

Day 2 3 0.0 8 ± 0.01 0.0 8 ± 0.01 0.0 8 ± 0.0 1 0.0 7 ± 0.0 1 0.0 8 ± 0.0 1 0.0 6 ± 0.0 1

Week 14 0.0 9 ± 0.0 1 0. 10 ± 0.0 1 0. 13 ± 0.01 0.0 9 ± 0.0 3 0. 12 ± 0.02 0.0 7 ± 0.02

Clinical Chemistry

n 10 10 10 10 10 10

Urea nitrogen (mg/dL)

Day 3 8.1 ± 0. 4 9 .0 ± 0. 2* 9.0 ± 0.4* 10.3 ± 0. 4 ** 11.0 ± 0. 5 ** 16.7 ± 0. 6* *

Day 2 3 10.7 ± 0.3 11.5 ± 0.6 11.5 ± 0.3 12.3 ± 0. 3** 14.6 ± 0. 5** 16.5 ± 0.4 **

Week 14 16. 9 ± 0. 3 15.8 ± 0. 5 15. 6 ± 0. 5* 16. 3 ± 0. 5 17. 0 ± 0. 3 14.5 ± 0.5 **

Creatinine (mg/dL)

Day 3 0. 24 ± 0.02 0. 26 ± 0.02 0. 26 ± 0.02 0. 26 ± 0.0 2 0. 26 ± 0.02 0. 30 ± 0.0 0 *

Day 2 3 0. 31 ± 0.0 1 0. 30 ± 0.0 0 0. 33 ± 0.02 0. 3 6 ± 0.0 2* 0. 37 ± 0.0 2** 0. 38 ± 0.01 **

Week 14 0. 61 ± 0.0 3 0.6 3 ± 0.0 2 0. 62 ± 0.02 0. 62 ± 0.0 1 0. 63 ± 0.02 0. 67 ± 0. 02

Total protein (g/dL)

Day 3 5.9 ± 0.1 5. 8 ± 0. 0 5. 7 ± 0.1 * 5. 6 ± 0.1* * 5. 6 ± 0.1** 5. 7 ± 0.1*

Day 2 3 6.3 ± 0. 0 6.2 ± 0.0 6.3 ± 0. 0 6. 3 ± 0.1 6. 2 ± 0.1 6. 2 ± 0.1

Week 14 7.3 ± 0.1 7.2 ± 0. 1 7.1 ± 0. 1 7.3 ± 0. 0 7.2 ± 0.1 7 .2 ± 0.1

Albumin (g/dL)

Day 3 4.3 ± 0.0 4.2 ± 0. 0 4.1 ± 0.0 ** 4. 1 ± 0. 0 ** 4.1 ± 0.0** 4. 2 ± 0. 0 **

Day 2 3 4.5 ± 0. 0 4. 4 ± 0.0 4.4 ± 0. 0 4. 4 ± 0.0 4. 3 ± 0. 0** 4. 3 ± 0.0 **

Week 14 4. 9 ± 0. 0 4. 8 ± 0.0 4. 7 ± 0. 1 4. 8 ± 0.0 4. 7 ± 0.0 4.8 ± 0.0

Globulin (g/dL)

Day 3 1.6 ± 0. 1 1.6 ± 0. 0 1.6 ± 0.0 1.5 ± 0. 0 1.5 ± 0. 0 1.6 ± 0. 0

Day 2 3 1.8 ± 0. 0 1.8 ± 0.0 1.9 ± 0. 0 1.8 ± 0.0 1.9 ± 0. 0 1.8 ± 0.0

Week 14 2.4 ± 0.1 2.5 ± 0.0 2.4 ± 0.0 2.5 ± 0.0 2.5 ± 0.0 2.5 ± 0.0

Albumin /globulin ratio

Day 3 2.7 ± 0. 1 2.7 ± 0. 0 2.7 ± 0. 1 2.7 ± 0. 0 2.7 ± 0. 1 2.7 ± 0. 0

Day 2 3 2 .5 ± 0. 0 2.4 ± 0.0 2.4 ± 0. 0 2.4 ± 0. 1 2.3 ± 0. 0** 2.4 ± 0.0 **

Week 14 2.0 ± 0. 0 2.0 ± 0.0 1.9 ± 0. 1 1.9 ± 0.0* 1.9 ± 0.0* 1.9 ± 0.0*

### 158 1- Bromopropane, NTP TR 564

T ABLE 

### F1

Hematology and Clinical Chemistry Data for Rats in the 3- Month Inhalation Study of 1

- Bromopropane

Chamber

Control 62.5 ppm 125 ppm 250 ppm 500 ppm 1, 0 00 ppm

M ale (continued)

Clinical Chemistry (continued)

n 10 10 10 10 10 10

Alanine aminotransferase (IU/L)

Day 3 56 ± 2 49 ± 1** 40 ± 1** 32 ± 1** 50 ± 7 ** 31 ± 5 **

Day 2 3 47 ± 1 44 ± 1 38 ± 1** 30 ± 1* * 25 ± 1 ** 22 ± 1 **

Week 14 141 ± 16 115 ± 9 121 ± 8 124 ± 9 120 ± 7 71 ± 5 **

Alkaline phosphatase (IU/L)

Day 3 689 ± 13 691 ± 1 5 679 ± 7 686 ± 1 2 706 ± 22 6 45 ± 1 8

Day 2 3 438 ± 1 1 467 ± 13 476 ± 9 470 ± 1 2 429 ± 9 453 ± 9

Week 14 258 ± 6 231 ± 3** 238 ± 6* 236 ± 4* 230 ± 5 ** 2 32 ± 5 **

Creatine kinase (IU/L)

Day 3 400 ± 43 b 385 ± 35 b 41 7 ± 37 b 397 ± 4 4 b 400 ± 22 b 311 ± 2 5 b

Day 2 3 417 ± 63 b 350 ± 41 b 380 ± 4 8 b 712 ± 190 b 363 ± 49 b 358 ± 39 b

Week 14 355 ± 113 b 191 ± 2 6 b 189 ± 4 0 b 22 4 ± 46 b 223 ± 38 b 137 ± 1 8 b

Sorbitol dehydrogenase (IU/L)

Day 2 3 17 ± 1 17 ± 0 17 ± 1 16 ± 1 18 ± 1 17 ± 0

Week 14 27 ± 2 24 ± 1 27 ± 1 30 ± 1 35 ± 1** 34 ± 1 **

Bile salts (μmol/L)

Day 3 19.5 ± 1.0 16.6 ± 0.5 16.4 ± 0.5 16.4 ± 0.9 25.3 ± 2.6 21.6 ± 2.2

Day 2 3 15.7 ± 0.4 15.0 ± 0.6 15.3 ± 0.3 15.2 ± 0.6 13.5 ± 0.4** 14.6 ± 0.5

Week 14 17.7 ± 0.9 15.7 ± 0.5 18.1 ± 1.2 16.4 ± 0.6 18.7 ± 0.6 17.5 ± 1.1



### Female

Hematology

n 10 10 10 10 10 10

Hematocrit (%)

Day 3 48.4 ± 0. 5 47.3 ± 0. 4 47. 7 ± 0. 5 47.7 ± 0. 5 47.8 ± 0. 5 47.4 ± 0.7

Day 2 3 47.6 ± 0. 5 48.1 ± 0.4 47.7 ± 0. 5 48.2 ± 0. 6 48.7 ± 0. 4 50.5 ± 0.4 **

Week 14 48.2 ± 0. 3 47.8 ± 0.4 47.6 ± 0. 5 48.8 ± 0. 3 47.3 ± 0.4 48.4 ± 0. 4

Packed cell volume ( mL/dL )

Day 3 48.2 ± 0. 4 47.2 ± 0. 4 46.9 ± 0.5 47.5 ± 0.6 47.7 ± 0. 6 47.8 ± 0. 6

Day 2 3 47.9 ± 0. 5 47.9 ± 0.4 47.5 ± 0. 3 48.4 ± 0.5 48.5 ± 0. 4 50.1 ± 0.4 **

Week 14 48.2 ± 0.4 47.6 ± 0.4 47.5 ± 0. 5 48.5 ± 0. 3 47.3 ± 0. 3 47.7 ± 0. 4

Hemoglobin (g/dL)

Day 3 15.0 ± 0. 2 14.5 ± 0.1 14.5 ± 0. 2 14.5 ± 0. 2 14.7 ± 0. 2 14.7 ± 0.2

Day 2 3 15.4 ± 0. 2 15.3 ± 0.1 15.4 ± 0. 1 15.6 ± 0.2 15.8 ± 0. 1* 16.2 ± 0. 2**

Week 14 15.7 ± 0.1 15.6 ± 0. 1 15.5 ± 0. 2 15.8 ± 0.1 15.6 ± 0.1 15.6 ± 0.1

Erythrocytes (10 6 /μL)

Day 3 7.97 ± 0. 11 7.69 ± 0.0 7 7.69 ± 0. 12 7.75 ± 0. 11 7.87 ± 0 .14 7.95 ± 0.11

Day 2 3 8.16 ± 0. 12 8.04 ± 0.0 6 8.03 ± 0. 06 8.08 ± 0. 11 8.23 ± 0. 10 8.48 ± 0.0 9

Week 14 8.39 ± 0.07 8.36 ± 0.0 6 8.34 ± 0. 09 8.49 ± 0.05 8.35 ± 0.0 4 8.45 ± 0.0 7

Reticulocytes (10 3 /μL)

Day 3 293.9 ± 16.6 295.4 ± 22.4 316.4 ± 10.9 274.5 ± 14.3 294.0 ± 14.6 290.2 ± 18.3

Day 2 3 156.7 ± 9.9 158.4 ± 11.8 148.5 ± 10.9 160.7 ± 8.2 162.3 ± 8.7 168.4 ± 7.6

Week 14 110.2 ± 11.5 114.7 ± 15.5 112.4 ± 18.5 113.0 ± 8.2 104.9 ± 17.8 115.9 ± 16.8

Nucleated erythrocytes /100 leukocytes

Day 3 0.20 ± 0.2 0 0. 20 ± 0. 13 0.20 ± 0. 13 0. 60 ± 0. 22 0. 60 ± 0. 31 0. 20 ± 0.1 3

Day 2 3 0. 20 ± 0. 1 3 0.00 ± 0.00 0. 20 ± 0. 13 0.1 0 ± 0.1 0 0.0 0 ± 0.0 0 0.1 0 ± 0.1 0

Week 14 0.70 ± 0. 21 0. 4 0 ± 0. 22 0.30 ± 0.1 5 0.50 ± 0. 27 0. 8 0 ± 0. 39 0.70 ± 0. 40

### 1- Bromopropane, NTP TR 564 159

T ABLE 

### F1

Hematology and Clinical Chemistry Data for Rats in the 3- Month Inhalation Study of 1

- Bromopropane

Chamber

Control 62.5 ppm 125 ppm 250 ppm 500 ppm 1, 0 00 ppm

Female (continued)

Hematology (continued)

n 10 10 10 10 10 10

Mean cell volume (fL)

Day 3 60.5 ± 0.5 61.4 ± 0. 2 61.0 ± 0. 4 61.3 ± 0. 2 60.6 ± 0. 4 60.2 ± 0.2

Day 2 3 58.8 ± 0. 4 59.5 ± 0.3 59.2 ± 0. 6 59.9 ± 0. 4 59.0 ± 0. 4 59.1 ± 0. 4

Week 14 57.4 ± 0.2 57.0 ± 0.2 56.9 ± 0.1 57.2 ± 0. 1 56.6 ± 0. 1** 56.5 ± 0. 2**

Mean cell hemoglobin (pg)

Day 3 18.8 ± 0.1 18.9 ± 0.1 18.8 ± 0.1 18.8 ± 0.1 18.7 ± 0.1 18.6 ± 0.1

Day 2 3 18.8 ± 0.1 19.1 ± 0.1 19.1 ± 0. 1 19.3 ± 0.1 * 19.2 ± 0. 2 19. 1 ± 0.1

Week 14 18. 7 ± 0.1 1 8.7 ± 0. 1 1 8.6 ± 0.1 18. 6 ± 0.1 18 .7 ± 0.1 18. 4 ± 0.1 *

Mean cell hemoglobin concentration (g/dL)

Day 3 3 1.1 ± 0.1 30.8 ± 0.1 30.8 ± 0.2 30.6 ± 0.1 30.8 ± 0. 2 30.8 ± 0.1

Day 2 3 32.1 ± 0.1 32.0 ± 0.1 32.4 ± 0.2 32.2 ± 0. 1 32.5 ± 0. 2 32.2 ± 0. 1

Week 14 32.6 ± 0.1 32.8 ± 0. 1 32.6 ± 0.1 32.6 ± 0.1 33.0 ± 0.1 32.6 ± 0. 1

Platelets (10 3 /μL)

Day 3 812.0 ± 25.7 824.1 ± 21.5 796.1 ± 19.3 805.1 ± 35.1 740.4 ± 30.7 778.1 ± 29.5

Day 2 3 773.3 ± 17.5 791.6 ± 13.7 806.4 ± 10.6 757.4 ± 19.3 791.4 ± 15.1 775.0 ± 21.8

Week 14 681.6 ± 12. 3 647.1 ± 10.7 656.6 ± 12.0 660.7 ± 15.7 658.2 ± 13.6 647.5 ± 11.6

Leukocytes (10 3 /μL)

Day 3 12.82 ± 0. 51 11.64 ± 0. 33 11.75 ± 0. 57 11.64 ± 0. 51 11.46 ± 0. 53 8.41 ± 0. 45**

Day 2 3 7.39 ± 0. 54 7.48 ± 0. 27 9.13 ± 0.3 9* 8.90 ± 0. 36 9.27 ± 0.4 1* 7.98 ± 0. 36

Week 14 6.34 ± 0.3 4 6.54 ± 0. 26 6.51 ± 0.3 1 6.55 ± 0. 31 6.44 ± 0.32 7.64 ± 0.4 3

Segmented neutrophils (10 3 /μL)

Day 3 1.41 ± 0.0 7 1. 43 ± 0. 07 1.29 ± 0.0 6 1.40 ± 0. 11 1.21 ± 0. 07* 1. 10 ± 0.1 4**

Day 2 3 1.10 ± 0.11 1. 09 ± 0. 08 1.33 ± 0.1 9 1.23 ± 0.1 5 1.28 ± 0. 14 1.12 ± 0.1 2

Week 14 1.03 ± 0.1 0 0.90 ± 0. 05 1.03 ± 0.0 6 1.10 ± 0. 09 0.89 ± 0. 05 1.33 ± 0. 10

Bands (10 3 /μL)

Day 3 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00

Day 2 3 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00

Week 14 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00

Lymphocytes (10 3 /μL)

Day 3 11.11 ± 0.50 9.97 ± 0. 35 10.20 ± 0. 54 9.98 ± 0.4 1 10.02 ± 0. 48 7.10 ± 0. 40**

Day 2 3 6.13 ± 0. 49 6.22 ± 0. 20 7.59 ± 0.3 1* 7.46 ± 0. 32 7.78 ± 0. 3 5 * 6.64 ± 0. 30

Week 14 5.04 ± 0. 28 5.49 ± 0. 24 5.25 ± 0. 28 5.19 ± 0. 27 5.29 ± 0.31 5.95 ± 0.3 8

Monocytes (10 3 /μL)

Day 3 0. 12 ± 0.0 1 0. 09 ± 0.0 2 0.1 2 ± 0.0 2 0. 11 ± 0.02 0. 10 ± 0.0 1 0. 08 ± 0.0 1

Day 2 3 0.05 ± 0.0 1 0. 07 ± 0.0 1 0. 07 ± 0.0 2 0. 07 ± 0.0 1 0.0 6 ± 0.0 1 0. 10 ± 0.0 2

Week 14 0.17 ± 0.06 0. 06 ± 0.0 2 0.13 ± 0.0 4 0.1 6 ± 0.0 5 0.1 6 ± 0.0 4 0. 29 ± 0.0 7

Basophils (10 3 /μL)

Day 3 0.0 18 ± 0.00 2 0.0 13 ± 0.00 3 0.0 15 ± 0.00 4 0.0 15 ± 0.00 3 0.0 13 ± 0.00 3 0.00 9 ± 0.00 2

Day 2 3 0.00 8 ± 0.00 7 0.00 5 ± 0.00 2 0.00 6 ± 0.00 2 0.00 6 ± 0.00 2 0.00 9 ± 0.00 2 0.00 6 ± 0.00 2

Week 14 0.00 4 ± 0.00 2 0.00 2 ± 0.00 1 0.0 15 ± 0.00 8 0.00 5 ± 0.00 2 0.00 7 ± 0.00 2 0.00 7 ± 0.00 3

Eosinophils (10 3 /μL)

Day 3 0. 17 ± 0.0 2 0.14 ± 0.01 0. 12 ± 0.01 0. 13 ± 0.02 0. 1 2 ± 0.01 0.1 2 ± 0.01

Day 2 3 0. 10 ± 0.0 1 0. 10 ± 0.0 1 0.1 3 ± 0.0 1 0.1 3 ± 0.0 1 0. 15 ± 0.02 * 0. 10 ± 0.0 1

Week 14 0.0 9 ± 0.0 1 0. 10 ± 0.0 1 0.0 8 ± 0.0 1 0.1 0 ± 0.01 0. 10 ± 0.02 0.0 7 ± 0.0 1

### 160 1- Bromopropane, NTP TR 564

T ABLE 

### F1

Hematology and Clinical Chemistry Data for Rats in the 3- Month Inhalation Study of 1

- Bromopropane

Chamber

Control 62.5 ppm 1 25 ppm 250 ppm 50 0 ppm 1, 0 00 ppm

Fem ale (continued)

Clinical Chemistry

n 10 10 10 10 10 10

Urea nitrogen (mg/dL)

Day 3 10.3 ± 0 .5 10.4 ± 0.5 8.9 ± 0. 5 10.2 ± 0.4 12.4 ± 0.4* 14.5 ± 0. 4 **

Day 2 3 11.6 ± 0. 4 11.4 ± 0.3 11.5 ± 0. 3 11.3 ± 0. 2 12.8 ± 0. 3** 16.6 ± 0. 3**

Week 14 16. 2 ± 0. 4 17.0 ± 0. 7 15.5 ± 0. 4 16.4 ± 0. 5 16.9 ± 0. 4 14.8 ± 0. 5

Creatinine (mg/dL)

Day 3 0.27 ± 0.0 2 0.25 ± 0.0 2 0.27 ± 0.0 2 0.27 ± 0.0 2 0.28 ± 0.0 1 0.31 ± 0.01

Day 2 3 0.24 ± 0.0 2 0. 27 ± 0.02 0. 28 ± 0.0 1 0.28 ± 0.0 1 0.30 ± 0.02 ** 0. 37 ± 0.0 2**

Week 14 0.70 ± 0.0 2 0.69 ± 0.01 0.68 ± 0.01 0.68 ± 0.01 0.63 ± 0.02 0.71 ± 0.0 2

Total protein (g/dL)

Day 3 6.1 ± 0.1 5.8 ± 0.1 ** 5. 9 ± 0.1 * 5. 8 ± 0.1 ** 5. 7 ± 0. 0** 5. 7 ± 0.1 **

Day 2 3 6.2 ± 0. 0 6.1 ± 0.0 6.3 ± 0.1 6.2 ± 0.1 6.2 ± 0.1 6.1 ± 0.1

Week 14 7.4 ± 0.1 7. 4 ± 0.1 7.3 ± 0.1 7.3 ± 0.1 7.4 ± 0.1 7.4 ± 0.1

Albumin (g/dL)

Day 3 4.6 ± 0.0 4. 4 ± 0.0 ** 4. 4 ± 0.0 ** 4. 4 ± 0.1 ** 4. 3 ± 0.0 ** 4. 3 ± 0. 0**

Day 2 3 4. 6 ± 0.0 4.5 ± 0.0 4.6 ± 0. 1 4.5 ± 0. 0 4.5 ± 0.0 4.5 ± 0. 0

Week 14 5.3 ± 0.0 5. 3 ± 0. 0 5. 2 ± 0.1 5.2 ± 0.1 5.3 ± 0. 0 5.2 ± 0. 1

Globulin (g/dL)

Day 3 1.5 ± 0.0 1.4 ± 0. 0 1.5 ± 0.0 1.4 ± 0. 0 1.4 ± 0.0 1.4 ± 0. 0

Day 2 3 1.7 ± 0. 0 1.6 ± 0.0 1.8 ± 0. 0 1.7 ± 0.0 1.7 ± 0. 0 1.6 ± 0.0

Week 14 2.1 ± 0. 0 2.1 ± 0.0 2.1 ± 0. 1 2.1 ± 0. 1 2.1 ± 0. 1 2.2 ± 0. 0

Albumin /globulin ratio

Day 3 3.2 ± 0. 1 3.1 ± 0.1 3.0 ± 0.0 3.1 ± 0.1 3.1 ± 0. 1 3.2 ± 0.1

Day 2 3 2.8 ± 0.1 2.9 ± 0. 1 2.6 ± 0. 0 2.7 ± 0.0 2.7 ± 0. 0 2.9 ± 0. 1

Week 14 2.6 ± 0. 0 2.5 ± 0.0 2.5 ± 0.0 2.6 ± 0.0 2.5 ± 0. 1** 2.4 ± 0.0* *

Alanine aminotransferase (IU/L)

Day 3 51 ± 2 41 ± 1 ** 33 ± 1** 26 ± 1 ** 25 ± 2 ** 19 ± 1 **

Day 2 3 44 ± 1 34 ± 1** 29 ± 1** 22 ± 1** 17 ± 1* * 20 ± 5 **

Week 14 71 ± 8 61 ± 5 57 ± 5 53 ± 3* 41 ± 4** 37 ± 7**

Alkaline phosphatase (IU/L)

Day 3 598 ± 15 589 ± 1 0 579 ± 1 7 563 ± 7 529 ± 13 ** 5 15 ± 11 **

Day 2 3 326 ± 9 342 ± 8 325 ± 9 326 ± 7 314 ± 9 337 ± 9

Week 14 215 ± 7 191 ± 5** 182 ± 5** 201 ± 6* 179 ± 5* * 187 ± 8**

Creatine kinase (IU/L)

Day 3 373 ± 33 b 329 ± 4 8 b 326 ± 25 b 338 ± 26 b 303 ± 29 b 217 ± 1 0 ** b

Day 2 3 285 ± 25 b 331 ± 25 b 334 ± 25 b 399 ± 57 b 311 ± 17 b 296 ± 1 8 b

Week 14 179 ± 20 b 182 ± 12 b 226 ± 37 b 168 ± 23 b 192 ± 34 b 157 ± 22 b

Sorbitol dehydrogenase (IU/L)

Day 2 3 16 ± 1 17 ± 0 15 ± 1 14 ± 2 16 ± 1 67 ± 27*

Week 14 19 ± 1 18 ± 1 18 ± 1 1 8 ± 1 18 ± 1 87 ± 33

Bile salts (μmol/L)

Day 3 17.3 ± 0.7 17.8 ± 1 .3 16.1 ± 0 .9 14.0 ± 0.8** 14.7 ± 0.7** 15.3 ± 0.9 *

Day 2 3 16.9 ± 1.0 13.6 ± 0.5 14.3 ± 0.9 13.1 ± 0.5* 13.3 ± 0.5* 19.2 ± 2.9

Week 14 16.2 ± 0.9 17.7 ± 1.2 15.9 ± 2.1 13.7 ± 0.5 15.7 ± 1.2 30.7 ± 6.3

* Significantly different (P ≤ 0.05) from the chamber control group by Dunn’s or Shirley’s test

** P ≤ 0.01

a Mean ± standard error. Statistical tests were performed on unrounded data.

b n=9

### 1- Bromopropane, NTP TR 564 161

T ABLE 

### F2

Hematology Data for Mice in the 3- Month Inhalation Study of 1

- Bromopropane a



### Chamber

Control 62.5 ppm 12 5 ppm 25 0 ppm 50 0 ppm

M ale

n 10 10 10 9 6

Hematocrit (%) 47.8 ± 0.4 48.4 ± 0. 3 49.3 ± 0. 4* 48. 1 ± 0. 2 47.9 ± 0. 5

Packed cell volume ( mL/dL ) 47.9 ± 0. 5 49.2 ± 0. 3 50.1 ± 0. 4* * 4 9

. 1 ± 0. 2 48.9 ± 0. 7

Hemoglobin (g/dL) 15.2 ± 0.2 1 5.5 ± 0. 1 15.9 ± 0. 1** 15.4 ± 0. 1 15.1 ± 0.2

Erythrocytes (10 6 /μL) 9.79 ± 0. 09 9.91 ± 0. 04 9.98 ± 0. 09 9.62 ± 0. 05 9.46 ± 0.1 5

Reticulocytes (10 3 /μL) 143.8 ± 8.9 158.6 ± 12.0 145.6 ± 12.6 130.2 ± 11.7 153.5 ± 26.9

Nucleated erythrocytes

/100 leukocytes 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00

Howell

- Jolly bodies

(%) erythrocytes 0.1 ± 0. 0 0.2 ± 0. 0 0.2 ± 0. 0 0.2 ± 0. 1 0.1 ± 0. 0

Mean cell volume (fL) 48.9 ± 0.2 49.6 ± 0.2 * 50.2 ± 0. 3** 51.1 ± 0. 2** 51.7 ± 0.2 **

Mean cell hemoglobin (pg) 15. 5 ± 0.1 15. 6 ± 0.1 15. 9 ± 0.1 ** 1 6.0 ± 0.1 ** 1 5 .9 ± 0.1* *

Mean cell hemoglobin

concentration (g/dL) 31.7 ± 0.1 31.6 ± 0. 1 31.6 ± 0.1 31.4 ± 0.1 * 30.9 ± 0. 1**

Platelets (10 3 /μL) 891.7 ± 27.6 894.0 ± 17.4 874.9 ± 14 .7 870.0 ± 1 5 .5 880.3 ± 43 .4

Leukocytes (10 3 /μL) 2.36 ± 0. 28 2.50 ± 0. 25 2.42 ± 0. 22 2.72 ± 0. 17 3.86 ± 0. 4 1 **

Segmented neutrophils (10 3 /μL) 0.29 ± 0.0 5 0.31 ± 0.0 3 0.29 ± 0.0 4 0.31 ± 0.0 3 0.43 ± 0.0 3*

Bands (10 3 /μL) 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00

Lymphocytes (10 3 /μL) 1.94 ± 0. 23 2.09 ± 0. 21 2.02 ± 0. 2 0 2.28 ± 0.1 6 3.31 ± 0. 36**

Monocytes (10 3 /μL) 0.06 ± 0.0 1 0.03 ± 0.0 1 0.0 4 ± 0.0 2 0.0 7 ± 0.02 0.0 5 ± 0.0 2

Basophils (10 3 /μL) 0.0 13 ± 0.00 2 0.00 9 ± 0.00 3 0.0 12 ± 0.00 3 0.00 9 ± 0.00 2 0.0 12 ± 0.00 3

Eosinophils (10 3 /μL) 0.06 ± 0.0 1 0.0 6 ± 0.0 2 0.05 ± 0.01 0.05 ± 0.0 1 0.0 7 ± 0.02



### Female

n 10 10 10 10 5

Hematocrit (%) 50.1 ± 0. 3 50.1 ± 0. 5 49.4 ± 0. 3 48.5 ± 0. 4** 48.6 ± 0. 5*

Packed cell volume ( mL/dL ) 50.9 ± 0. 3 50.7 ± 0. 4 50.0 ± 0. 2 49.1 ± 0. 5** 49.1 ± 0. 5*

Hemoglobin (g/dL) 16.2 ± 0. 1 16.2 ± 0.1 15. 9 ± 0. 1 15. 6 ± 0.1 ** 15. 6 ± 0.2 *

Erythrocytes (10 6 /μL) 10.18 ± 0. 08 10.13 ± 0.0 7 9.92 ± 0.0 4 * 9.60 ± 0.0 8** 9.53 ± 0.1 1**

Reticulocytes (10 3 /μL) 153.9 ± 1 0.6 148.8 ± 10.9 134.9 ± 17.3 123.9 ± 8.0 112.2 ± 10.1

Nucleated erythrocytes

/ 100 leukocytes 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00

Howell

- Jolly bodies

(%) erythrocytes 0.0 ± 0. 0 0.1 ± 0. 0 0.1 ± 0. 0 0.1 ± 0. 0 0.0 ± 0. 0

Mean cell volume (fL) 50.0 ± 0.2 50.1 ± 0.1 50.4 ± 0.2 51 .1 ± 0.2 ** 51.6 ± 0. 2**

Mean cell hemoglobin (pg) 15. 9 ± 0.1 1 6.0 ± 0. 0 16.1 ± 0.1 16 .3 ± 0.1 ** 1 6.3 ± 0.1 **

Mean cell hemoglobin

concentration (g/dL) 31.8 ± 0. 1 32.0 ± 0.1 31.9 ± 0.1 31.8 ± 0.1 31.7 ± 0. 2

Platelets (10 3 /μL) 846.7 ± 18.9 821.5 ± 30.0 835.0 ± 17.5 819.0 ± 16.9 732.4 ± 5 0.0

Leukocytes (10 3 /μL) 2.73 ± 0. 24 2.81 ± 0.2 3 2.33 ± 0. 16 2.78 ± 0. 33 2.57 ± 0. 25

Segmented neutrophils (10 3 /μL) 0.32 ± 0.0 5 0.29 ± 0. 03 0.26 ± 0.0 2 0.31 ± 0.0 3 0.26 ± 0. 03

Bands (10 3 /μL) 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00

Lymphocytes (10 3 /μL) 2.35 ± 0. 1 9 2.45 ± 0. 21 1.97 ± 0. 14 2.36 ± 0.3 2 2.25 ± 0. 22

Monocytes (10 3 /μL) 0.0 4 ± 0.01 0.0 3 ± 0.01 0.0 5 ± 0.0 2 0.0 8 ± 0.0 2 0.0 4 ± 0.02

Basophils (10 3 /μL) 0.00 7 ± 0.00 2 0.0 1 0 ± 0.00 2 0.00 9 ± 0.00 3 0.0 12 ± 0.00 2 0.00 8 ± 0.00 4

Eosinophils (10 3 /μL) 0.0 2 ± 0.0 1 0.0 3 ± 0.01 0.0 4 ± 0.0 1 0.0 3 ± 0.0 0 0. 0 1 ± 0.0 0

* Significantly different (P ≤ 0.05) from the chamber control group by Dunn’s or Shirley’s test

** P ≤ 0.0 1

a Mean ± standard error. Statistical tests were performed on unrounded data.

### 162 1- Bromopropane, NTP TR 564

### 163



# APPENDIX G

ORGAN WEIGHTS

AND ORGAN

- WEIGHT

- TO

- BODY

- WEIGHT RATIOS

T ABLE 

### G1 Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios for Rats

in the 2- Week Inhalation Study of 1- Bromopropane

...................................................................... 164

T ABLE 

### G2 Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios for Rats

in the 3- Month Inhalation Study of 1- Bromopropane

................................................................... 165

T ABLE 

### G3 Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios for Mice

in the 2- Week Inhalation Study of 1- Bromopropane

..................................................................... 166

T ABLE 

### G4 Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios for Mice

in the 3- Month Inhalation Study of 1- Bromopropane

................................................................... 167

### 164 1- Bromopropane, NTP TR 564

T ABLE 

### G1

Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios for Rats in the 2- Week Inhalation Study

of

1- Bromopropane a



### Chamber

Control 1 25 ppm 250 ppm 500 ppm 1 ,000 ppm 2 ,000 ppm

M ale

n 5 5 5 4 5 5

Necropsy body wt 169 ± 4 167 ± 4 169 ± 5 166 ± 6 164 ± 5 123 ± 4**

Heart

Absolute 0.62 ± 0.0 1 0.6 1 ± 0.01 0.63 ± 0.0 2 0.63 ± 0.02 0. 6 2 ± 0.0 2 0. 56 ± 0.0 2

Relative 3.686 ± 0.0 26 3. 648 ± 0.11 6 3. 711 ± 0.0 3 2 3. 779 ± 0.07 5 3. 802 ± 0. 046 4.550 ± 0.1 03**

R. Kidney

Absolute 0. 65 ± 0.0 2 0. 70 ± 0.0 2 0.7 0 ± 0.0 2 0. 72 ± 0.03 0. 79 ± 0.03** 0. 63 ± 0.0 2

Relative 3.843 ± 0. 096 4.191 ± 0.0 76* 4. 162 ± 0. 07 4 * 4.3 50 ± 0.0 95** 4. 813 ± 0. 091** 5.144 ± 0. 090**

Liver

Absolute 7.57 ± 0. 14 7.26 ± 0. 17 7.61 ± 0.2 2 7.98 ± 0. 22 8.93 ± 0. 19** 6.57 ± 0. 24**

Relative 44.895 ± 0.729 43.588 ± 0. 572 45.158 ± 0.735 48.215 ± 0. 754** 54.535 ± 0.909** 53.600 ± 0.936**

Lung

Absolute 1.37 ± 0. 15 1.36 ± 0.1 3 1.26 ± 0. 11 1.16 ± 0.11 1.10 ± 0.0 5 0.9 0 ± 0.0 5 **

Relative 8.122 ± 0. 878 8.241 ± 0. 991 7.449 ± 0. 556 7.001 ± 0. 617 6.674 ± 0. 131 7.365 ± 0. 314

R. Testis

Absolute 1.000 ± 0.0 27 1. 022 ± 0.0 19 1. 018 ± 0.03 3 1.0 16 ± 0.0 46 1.022 ± 0.0 37 0.893 ± 0.0 42

Relative 5.924 ± 0. 075 6.1 42 ± 0.0 87 6.0 36 ± 0.1 00 6. 127 ± 0.1 02 6.229 ± 0.1 42 7. 265 ± 0.1 19 **

Thymus

Absolute 0.461 ± 0.02 5 0.4 57 ± 0.0 16 0. 477 ± 0.0 28 0. 523 ± 0.01 7 0. 452 ± 0.01 6 0. 286 ± 0.01 1 **

Relative 2. 742 ± 0.1 89 2. 743 ± 0.0 87 2. 826 ± 0.1 42 3.177 ± 0. 200 2.761 ± 0. 104 2.342 ± 0. 103



### Female

n 5 5 5 5 5 5

Necropsy body wt 120 ± 2 124 ± 3 120 ± 4 124 ± 5 119 ± 2 107 ± 2*

Heart

Absolute 0.48 ± 0.01 0.48 ± 0.01 0.4 8 ± 0.01 0. 5 4 ± 0.01** 0.48 ± 0.01 0.51 ± 0.01

Relative 3. 984 ± 0.0 68 3. 877 ± 0.1 43 3. 982 ± 0. 081 4.325 ± 0.06 9 4.058 ± 0.0 36 4.759 ± 0. 103**

R. Kidney

Absolute 0. 48 ± 0.0 1 0.58 ± 0.0 1** 0.5 4 ± 0.02 ** 0.57 ± 0.0 3** 0.6 0 ± 0.0 2 ** 0.6 1 ± 0.0 2 **

Relative 3.999 ± 0.0 74 4. 647 ± 0. 116** 4. 521 ± 0. 036** 4.6 15 ± 0. 067** 5.049 ± 0.1 07** 5.718 ± 0.16 1**

Liver

Absolute 4.75 ± 0. 1 2 5.24 ± 0.09 5.22 ± 0.2 8 6.01 ± 0.2 6** 6.19 ± 0.1 9 ** 5.82 ± 0.1 3**

Relative 39.548 ± 0.581 42.259 ± 0.673* 43.253 ± 0.977** 48.342 ± 0.856 ** 52.105 ± 1.280 ** 54.698 ± 0.8 3 2* *

Lung

Absolute 0.83 ± 0.0 3 1.07 ± 0.0 9* 0.91 ± 0.04 1.22 ± 0. 11 0. 8 6 ± 0.0 4 0.79 ± 0.0 2

Relative 6.915 ± 0.2 0 1 8.609 ± 0. 579 7.586 ± 0.3 77 9.874 ± 1.027** 7.245 ± 0. 304 7.395 ± 0. 258

Thymus

Absolute 0.3 60 ± 0.00 7 0. 400 ± 0.0 12 0.365 ± 0.0 22 0. 424 ± 0.0 31 0. 374 ± 0.02 3 0. 300 ± 0.01 3

Relative 3.000 ± 0. 023 3.220 ± 0.09 5 3.025 ± 0. 086 3.417 ± 0.2 03 3.138 ± 0. 155 2.817 ± 0.1 1 6

* Significantly different (P ≤ 0.05) from the chamber control group by Williams’ or Dunnett’s test

** P ≤ 0.01

a Organ weights (absolute weights) and body weights are given in grams; organ

- weight

- to

- body

- weight ratios (relative weights) are given as

mg organ weight/g body weight (mean ± standard error).

### 1- Bromopropane, NTP TR 564 165

T ABLE 

### G2

Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios for Rats in the 3

- Month Inhalation Study

of

1- Bromopropane a



### Chamber

Control 62.5 ppm 12 5 ppm 25 0 ppm 50 0 ppm 1, 0 00 ppm

n 10 10 10 10 10 10



### M ale

Necropsy body wt 345 ± 7 346 ± 5 348 ± 6 350 ± 7 344 ± 10 305 ± 6 **

Heart

Absolute 0.93 ± 0.0 2 0.93 ± 0.02 0.9 5 ± 0.0 1 0.9 3 ± 0.01 0. 92 ± 0.0 2 0. 8 6 ± 0.0 2 *

Relative 2.690 ± 0.0 23 2.698 ± 0.0 27 2.727 ± 0.03 8 2.662 ± 0.0 37 2.675 ± 0.05 9 2.819 ± 0.0 34

R. Kidney

Absolute 1.0 1 ± 0.0 2 1.0 2 ± 0.02 1.0 6 ± 0.02 1.0 8 ± 0.0 3 1.08 ± 0.0 7 1.02 ± 0.0 3

Relative 2.929 ± 0.0 24 2.936 ± 0.04 1 3.035 ± 0.0 19 3.102 ± 0. 110 3.161 ± 0. 209 3.3 47 ± 0.05 0 **

Liver

Absolute 11.21 ± 0.2 8 11.21 ± 0. 30 11.71 ± 0.3 3 12.45 ± 0. 31* 13.34 ± 0. 43** 13.23 ± 0. 36 **

Relative 32.435 ± 0. 387 32.318 ± 0. 481 33.674 ± 0. 638 35.543 ± 0. 696 ** 38.800 ± 0. 435 ** 43.386 ± 0. 496 **

Lung

Absolute 1. 73 ± 0.06 2 .01 ± 0. 18 1. 82 ± 0.0 8 1.80 ± 0.0 4 1. 69 ± 0.0 5 1. 55 ± 0.0 5

Relative 5.007 ± 0.1 84 5. 797 ± 0. 4 65 5.2 57 ± 0. 248 5. 152 ± 0. 165 4.950 ± 0.2 02 5.1 01 ± 0. 123

Spleen

Absolute 0.678 ± 0.01 5 0.682 ± 0.0 12 0.677 ± 0.0 15 0.689 ± 0.01 4 0.691 ± 0.02 1 0.625 ± 0.0 15

Relative 1.963 ± 0.0 24 1.970 ± 0.01 7 1.947 ± 0.0 27 1.968 ± 0.0 31 2.011 ± 0. 030 2.053 ± 0.0 33

R. Testis

Absolute 1.4 3 0 ± 0.0 27 1.4 34 ± 0.02 2 1.4 3 6 ± 0.0 34 1.4 05 ± 0.0 22 1.3 20 ± 0.0 78 1.3 36 ± 0.0 17

Relative 4. 142 ± 0.0 50 4.1 47 ± 0.0 77 4. 132 ± 0.0 79 4 .015 ± 0.04 4 3.809 ± 0.1 70 4.396 ± 0.0 69

Thymus

Absolute 0. 346 ± 0.01 2 0.3 21 ± 0.0 10 0.32 7 ± 0.01 2 0.3 38 ± 0.0 15 0. 355 ± 0.01 6 0. 323 ± 0.01 5

Relative 1. 002 ± 0.0 33 0.927 ± 0.02 8 0.941 ± 0.0 28 0.965 ± 0.0 42 1.0 29 ± 0. 02 1 1. 064 ± 0.05 0



### Female

Necropsy body wt 202 ± 3 203 ± 5 207 ± 4 209 ± 2 205 ± 3 190 ± 4

Heart

Absolute 0. 59 ± 0.01 0.6 0 ± 0.0 1 0.6 2 ± 0.01 0. 63 ± 0.01 0. 63 ± 0.01 0.6 1 ± 0.01

Relative 2.942 ± 0.0 64 2.978 ± 0.0 53 3.009 ± 0.0 59 3.001 ± 0.02 8 3.090 ± 0.05 8 3.207 ± 0.0 5 1 **

R. Kidney

Absolute 0.6 5 ± 0.01 0.6 6 ± 0.0 2 0.67 ± 0.01 0.6 8 ± 0.01 0. 68 ± 0.01 0. 71 ± 0.0 2 **

Relative 3.209 ± 0.0 37 3.237 ± 0.0 43 3.252 ± 0.0 81 3.263 ± 0.0 45 3.298 ± 0.0 45 3.719 ± 0.05 8**

Liver

Absolute 6.02 ± 0.0 9 6.38 ± 0.2 0 6.62 ± 0.1 6* 6. 60 ± 0.1 8* 7.29 ± 0.1 7** 7.98 ± 0. 1 8 * *

Relative 29.910 ± 0. 219 31.398 ± 0.5 38 32.001 ± 0.5 73 * 31.497 ± 0. 754* 35.490 ± 0. 666** 41.895 ± 0.39 8* *

Lung

Absolute 1. 16 ± 0.0 3 1.1 9 ± 0.0 3 1.3 0 ± 0.0 6 1.2 4 ± 0.0 5 1. 25 ± 0.0 5 1. 08 ± 0.0 3

Relative 5.752 ± 0.1 26 5.904 ± 0. 199 6.283 ± 0.2 76 5.945 ± 0.2 32 6.089 ± 0. 252 5.658 ± 0. 172

Spleen

Absolute 0.4 20 ± 0.00 9 0. 422 ± 0.010 0. 446 ± 0.0 06* 0. 461 ± 0.00 5 * 0. 451 ± 0.00 9 * 0. 439 ± 0.0 10 *

Relative 2.085 ± 0.0 30 2.084 ± 0.0 53 2.162 ± 0.04 8 2.204 ± 0.0 30 2.200 ± 0.0 52 2.311 ± 0.0 57**

Thymus

Absolute 0.2 61 ± 0.00 9 0.271 ± 0.00 8 0.2 90 ± 0.01 2 0.3 01 ± 0.0 11 * 0.3 02 ± 0.0 1 0* 0.2 71 ± 0.0 14

Relative 1. 296 ± 0.04 2 1. 336 ± 0.0 39 1.39 7 ± 0.0 3 9 1.4 37 ± 0.0 4 2 1.4 71 ± 0.0 50* 1.4 25 ± 0.0 73*

* Significantly different (P ≤ 0.05) from the chamber control group by Williams’ or Dunnett’s test

** (P ≤ 0.01 )

a Organ weights (absolute weights) and body weights are given in grams; organ

- weight

- to

- body

- weight ratios (relative weights) are given as

mg organ weight/g body weight (mean ± standard error).

### 166 1- Bromopropane, NTP TR 564

T ABLE 

### G3

Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios for Mice in the 2- Week Inhalation Study

of

1- Bromopropane a



### Chamber

Control 1 25 ppm 250 ppm 500 ppm 1 ,000 ppm 2 ,000 ppm

M ale

n 5 5 5 1 4 0 b

Necropsy body wt 28.0 ± 0.4 28.9 ± 0. 5 27.0 ± 0. 4 25.8 26.6 ± 0.5

Heart

Absolute 0.1 4 ± 0.0 1 0.1 4 ± 0.0 1 0.1 2 ± 0.00 0.1 2 0. 11 ± 0.0 1 **

Relative 4 .995 ± 0. 216 4.906 ± 0.1 49 4.594 ± 0.0 75 4.651 4.243 ± 0.23 2 *

R. Kidney

Absolute 0.24 ± 0.01 0.25 ± 0.0 1 0.2 4 ± 0.00 0. 24 0. 2 4 ± 0.0 1

Relative 8.641 ± 0. 099 8.638 ± 0.30 1 8.743 ± 0. 127 9.302 8.9 41 ± 0. 161

Liver

Absolute 1.46 ± 0.0 4 1.50 ± 0.0 5 1.39 ± 0.0 2 1.44 1.65 ± 0.05 *

Relative 52.109 ± 0. 653 51.841 ± 0. 99 2 51.568 ± 0. 464 55.814 61.936 ± 1.839 **

Lung

Absolute 0.1 8 ± 0.01 0.1 9 ± 0.0 1 0.1 9 ± 0.00 0.1 9 0.1 8 ± 0.0 1

Relative 6.562 ± 0. 238 6.489 ± 0.2 91 7.116 ± 0. 111 7.364 6.679 ± 0. 125

R. Testis

Absolute 0.10 7 ± 0.00 2 0.10 2 ± 0.00 5 0. 10 7 ± 0.002 0. 1 09 0.1 03 ± 0.00 3

Relative 3 .807 ± 0. 029 3.516 ± 0. 162 3.955 ± 0. 127 4.2 25 3.863 ± 0.1 4 9

Thymus

Absolute 0.0 59 ± 0.00 2 0.0 57 ± 0.00 4 0.0 60 ± 0.00 3 0.0 49 0.0 50 ± 0.00 6

Relative 2 .107 ± 0. 080 1.9 60 ± 0.15 0 2.2 22 ± 0. 109 1. 899 1.876 ± 0.1 91



### Female

n 5 5 5 5 4 3

Necropsy body wt 22.9 ± 0. 2 22.2 ± 0. 5 22.8 ± 0. 2 23.4 ± 0.6 22.8 ± 0. 5 22.7 ± 0. 3

Heart

Absolute 0.12 ± 0.00 0.1 2 ± 0.00 0.1 2 ± 0.0 1 0. 12 ± 0.0 1 0. 11 ± 0.0 1 0. 12 ± 0.00

Relative 5.1 46 ± 0. 129 5.410 ± 0.0 82 5.258 ± 0. 256 4. 990 ± 0. 314 4.923 ± 0. 161 5.152 ± 0.2 05

R. Kidney

Absolute 0.16 ± 0.00 0.17 ± 0.00 0.18 ± 0.00 * 0.1 7 ± 0.0 1 * 0.1 9 ± 0.0 1 ** 0.1 9 ± 0.00**

Relative 6.977 ± 0.1 35 7.762 ± 0.1 74 7. 795 ± 0.1 69 7. 377 ± 0. 315 8.355 ± 0.4 25* * 8.235 ± 0. 03 4* *

Liver

Absolute 1.19 ± 0.03 1. 1 3 ± 0.0 3 1. 23 ± 0. 02 1. 35 ± 0.03 * 1.52 ± 0.0 7** 1.69 ± 0. 1 0 **

Relative 51.985 ± 0 .718 51.014 ± 0.533 53.680 ± 0 .826 57.791 ± 1.0 92 ** 66.739 ± 2.104 ** 74.471 ± 3.176 **

Lung

Absolute 0.1 7 ± 0.00 0.1 8 ± 0.0 1 0.1 9 ± 0.0 1 0.1 9 ± 0.01 0.1 9 ± 0.0 1 0.20 ± 0.0 1

Relative 7. 506 ± 0.1 63 8.287 ± 0.25 9 8.404 ± 0. 220 8. 147 ± 0.370 8.348 ± 0. 330 8.677 ± 0. 274*

Thymus

Absolute 0.07 4 ± 0.00 4 0.0 7 8 ± 0.004 0.0 77 ± 0.00 2 0.0 62 ± 0.00 6 0.0 58 ± 0.00 5* 0.0 48 ± 0.00 5 **

Relative 3. 208 ± 0.1 69 3. 527 ± 0. 1 1 1 3. 389 ± 0. 0 8 0 2.689 ± 0. 319 2.552 ± 0.2 62 * 2.114 ± 0.1 99 **

* Significantly different (P ≤ 0.05) from the chamber control group by Williams’ or Dunnett’s test

** P ≤ 0.01

a Organ weights (absolute weights) and body weights are given in grams; organ

- weight

- to

- body

- weight ratios (relative weights) are given as

mg organ weight/g body weight (mean ± standard error).

b No data were available for the 2, 000 ppm male s due to 100% mortality.

### 1- Bromopropane, NTP TR 564 167

T ABLE 

### G4

Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios for Mice in the 3

- Month Inhalation Study

of

1- Bromopropane a



### Chamber

Control 62.5 ppm 125 ppm 250 ppm 50 0 ppm

M ale

n 10 10 10 9 6

Necropsy body wt 39.6 ± 0. 8 3 9.0 ± 0. 9 39.6 ± 0. 6 37.9 ± 0. 7 37.4 ± 1.4

Heart

Absolute 0.16 ± 0.00 0.1 6 ± 0.00 0.16 ± 0.00 0.15 ± 0.00 0.1 6 ± 0.00

Relative 4. 051 ± 0.0 54 4. 014 ± 0. 050 4. 023 ± 0.0 49 4. 080 ± 0. 058 4. 210 ± 0.1 48

R. Kidney

Absolute 0. 3 2 ± 0.0 1 0.3 1 ± 0.0 1 0. 30 ± 0.00 0.2 9 ± 0.0 1** 0.2 8 ± 0.0 1 **

Relative 8. 133 ± 0. 216 7.947 ± 0.123 7.669 ± 0.1 17 7.687 ± 0. 208 7.458 ± 0. 258*

Liver

Absolute 1.68 ± 0.0 5 1.66 ± 0.0 5 1.7 1 ± 0.03 1.71 ± 0.0 3 1.80 ± 0.0 8

Relative 42.485 ± 0.50 3 42.584 ± 0.6 41 43.288 ± 0. 379 45.129 ± 0. 393 ** 48.038 ± 1

. 126 **

Lung

Absolute 0.2 1 ± 0.01 0.2 1 ± 0.01 0.2 3 ± 0.01 0.2 1 ± 0.0 0 0.2 1 ± 0.01

Relative 5.236 ± 0. 147 5.477 ± 0. 168 5.734 ± 0.2 59 5.505 ± 0.1 67 5.742 ± 0.3 02

Spleen

Absolute 0. 067 ± 0.00 2 0.067 ± 0.002 0.0 69 ± 0.00 2 0. 067 ± 0.00 2 0. 06 3 ± 0.00 2

Relative 1.696 ± 0.0 51 1.724 ± 0.0 5 0 1.743 ± 0. 032 1.763 ± 0.0 51 1.699 ± 0.0 69

R. Testis

Absolute 0.11 6 ± 0.00 3 0.1 20 ± 0.002 0.10 8 ± 0.00 9 0.11 4 ± 0.00 2 0.11 5 ± 0.00 1

Relative 2.933 ± 0.0 53 3. 099 ± 0.0 51 2.747 ± 0. 22 6 3.014 ± 0.0 5 0 3.081 ± 0.0 98

Thymus

Absolute 0.04 9 ± 0.00 5 0.04 5 ± 0.00 4 0.04 7 ± 0.00 4 0.04 9 ± 0.003 0.0 56 ± 0.00 6

Relative 1. 236 ± 0. 107 1. 154 ± 0.0 84 1.19 2 ± 0.0 87 1. 277 ± 0.0 60 1. 481 ± 0. 104



### Female

n 10 10 10 10 5

Necropsy body wt 30.9 ± 0.7 33.0 ± 1.3 31.9 ± 0. 8 30.2 ± 0. 9 31.3 ± 1.4

.

Heart

Absolute 0.1 4 ± 0.00 0.14 ± 0.00 0.1 4 ± 0.00 0.1 4 ± 0.0 1 0.1 4 ± 0.0 1

Relative 4. 3 78 ± 0. 087 4.33 2 ± 0.1 12 4. 404 ± 0.07 8 4.6 51 ± 0.11 0 4. 365 ± 0. 210

R. Kidney

Absolute 0. 20 ± 0.00 0. 2 0 ± 0.00 0. 2 1 ± 0.0 1 0. 20 ± 0.0 1 0. 22 ± 0.0 1* *

Relative 6. 330 ± 0.1 55 6. 199 ± 0.1 79 6. 552 ± 0.1 84 6.780 ± 0. 164 7.187 ± 0. 265**

Liver

Absolute 1. 42 ± 0.0 3 1. 49 ± 0.0 6 1. 51 ± 0.03 1. 4 9 ± 0.04 1. 65 ± 0. 11*

Relative 46.102 ± 0. 767 45.104 ± 0. 424 47.586 ± 0.8 8 0 49.377 ± 0. 6 40 ** 52.666 ± 1. 904 **

Lung

Absolute 0.2 0 ± 0.01 0.22 ± 0.01 0.2 2 ± 0.0 1 0.21 ± 0.01 0.2 6 ± 0.0 2**

Relative 6.518 ± 0. 276 6.642 ± 0. 298 7.018 ± 0.2 22 7.028 ± 0. 190 8.497 ± 0.9 2 9 **

Spleen

Absolute 0. 093 ± 0.00 2 0. 098 ± 0.00 3 0.0 96 ± 0.00 4 0. 092 ± 0.00 4 0. 082 ± 0.00 7

Relative 3.0 19 ± 0.0 73 2 .980 ± 0.07 2 3. 006 ± 0. 079 3. 060 ± 0. 114 2.643 ± 0. 257

Thymus

Absolute 0.05 8 ± 0.00 5 0.05 8 ± 0.003 0.05 4 ± 0.002 0.0 5 4 ± 0.00 2 0.05 5 ± 0.00 4

Relative 1. 861 ± 0.1 45 1. 749 ± 0.0 81 1. 692 ± 0.07 3 1. 797 ± 0.0 62 1.767 ± 0. 138

* Significantly different (P ≤ 0.05) from the chamber control group by Williams’ or Dunnett’s test

** P ≤ 0.01

a Organ weights (absolute weights) and body weights are given in grams; organ

- weight

- to

- body

- weight ratios (relative weights) are given as

mg organ weight/g body weight (mean ± standard error).

### 168 1- Bromopropane, NTP TR 564

### 169



# APPENDIX H

REPRODUCTIVE TISSUE EVALUATIONS

AND ESTROUS CYCLE CHARAC TERIZATION

T ABLE 

### H1 Summary of Reproductive Tissue Evaluations for Male Rats

i n the 3- Month Inhalation Study of 1- Bromopropane

.................................................................... 170

T ABLE 

### H2 Estrous Cycle Characterization for Female Rats

in the 3- Month Inhalation Study of 1- Bromopropane

................................................................... 170

T ABLE 

### H3 Summary of Reproductive Tissue Evaluations for Male Mice

i n the 3- Month Inhalation Study of 1

- Bromopropane

................................................................... 171

T ABLE 

### H4 Estrous Cycle Characterization for Female Mice

in the 3- Month Inhalation Study of 1

- Bromopropane

................................................................... 171

### 170 1- Bromopropane, NTP TR 564

T ABLE 

### H1

Summary of Reproductive Tissue Evaluations for Male Rats in the 3- Month Inhalation St udy

o f 1

- Bromopropane a



### Chamber Control 250 ppm 500 ppm 1,000 ppm

n 10 10 10 10

Weights (g)

Necropsy body wt 3 45 ± 7 35 0 ± 7 344 ± 10 305 ± 6 **

L. Cauda epididymis 0.1865 ± 0.00 39 0. 1929 ± 0.00 38 0. 1740 ± 0.0 106 0. 1603 ± 0.00 67*

L. Epididymis 0. 4862 ± 0.00 81 0.4 99 4 ± 0.0 042 0.4 622 ± 0.01 92 0.4 423 ± 0.00 83*

L. Testis 1.5128 ± 0.0 313 1. 4833 ± 0.0 252 1.38 67 ± 0.0 839 1.4 104 ± 0.026 9

Spermatid measurement

Spermatid heads (10 6 /g testis) 123.6 ± 2.3 129.9 ± 6 .1 112.9 ± 12.3 130.5 ± 3.8

Spermatid heads (10 6 /testis) 171.8 ± 4.6 172.0 ± 9.2 150.8 ± 16.9 169.5 ± 6.0

Epididymal spermatozoal measurements

M otility (%) 90.99 ± 0.83 84.87 ± 0.94** 81.84 ± 0.80** b 65.82 ± 3.17**

Sperm (10 6 /g cauda epididymis) 699 ± 26 638 ± 34 576 ± 63 523 ± 3 0**

Sperm (10 6 /cauda epididymis) 130.27 ± 5.19 122.90 ± 6.63 105.27 ± 1 2 .01 82.39 ± 2.51**

* Significantly different ( P ≤ 0.0 5) from the chamber control group by Williams’ test ( cauda epididymis and epididymis weights)

** Significantly different (P ≤ 0.01) from the chamber control group by Williams’ test (body weights) or Shirley’s test (epididymal spermatozoa l

measurements)

a Data are presented as mean ± standard error. Differences from the chamber control group are not significant by Dunnett’s test (te stis weights)

or Dunn’s test (spermatid measurements).

b n=9



### T ABLE 

### H2

Estrous Cycle Characterization for Female Rats in th e 3- Month Inhalation Study of 1

- Bromopropane a



### Chamber Control 250 ppm 500 ppm 1,000 ppm

Number weighed at necropsy 10 10 10 10

Necropsy body wt (g) 202 ± 3 209 ± 2 205 ± 3 1 90 ± 4

Proportion of regular cycling females b 9/ 10 10 /10 10 / 10 10/10

Estrous cycle length (days) 4. 9 ± 0. 11 c 5.0 ± 0. 05 4. 9 ± 0. 07 5.0 ± 0. 05

Estrous stages (% of cycle) d

Diestrus 55.8 43.3 40.0 41.7

Proestrus 17.5 5 .0 4.2 6.7

Estrus 20 .8 3 2.5 3 6.7 3 3.3

Metestrus 5. 0 1 9.2 1 9.2 18.3

Uncertain diagnoses 0. 8 0.0 0. 0 0.0

a Necropsy body weights and estrous cycle length data are presented as mean ± standard error. Differences from the chamber con trol group are

not significant by Dunnett’s test (body weight) or Dunn’s test (estrous cycle length).

b Number of females with a regular cycle/number of females cycling

c Estrous cycle was longer than 12 days or unclear in 1 of 10 animals.

d Evidence shows that females exposed to 250, 500, or 1,000 ppm differ significantly (Wilk’s Criterion, P ≤ 0.05) from the chamber control

females in the relative length of time spent in the estrous stages. The tests for equality of transition probability matrices among exposed

groups and between the chamber control group and each exposed group indicated th at e xposed females spent significantly more time in

extended estrus (P<0.001) and significantly less time in extended diestrus (P<0.005) than the chamber control group.

### 1- Bromopropane, NTP TR 564 171

T ABLE 

### H3

Summary of Reproductive Tissue Evaluations for Male Mice in the 3- Month Inhalation St udy

o f 1

- Bromopropane a



### Chamber Control 125 ppm 250 ppm 500 ppm

n 10 10 9 6

Weights (g)

Necropsy body wt 39.6 ± 0.8 39.6 ± 0.6 37.9 ± 0.7 37.4 ± 1.4

L. Cauda epididymis 0. 0164 ± 0.00 08 0. 0170 ± 0.00 09 0. 0179 ± 0.0 0 05 0. 0192 ± 0.00 07

L. Epididymis 0. 0538 ± 0.00 10 0. 0492 ± 0.0 0 16* 0. 0532 ± 0.0 014 0. 0535 ± 0.00 12

L. Testis 0.1137 ± 0.0 020 0.1063 ± 0. 0085 0.1104 ± 0.0 028 0.1103 ± 0.0 010

Spermatid measurement

Spermatid heads (10 6 /g testis) 188.7 ± 8.1 164.9 ± 19.7 180.5 ± 3 .7 199.3 ± 8 .0

Spermatid heads (10 6 /testis) 19.83 ± 0.89 17.57 ± 2.09 18.17 ± 0.48 20.11 ± 0 .59

Epididymal spermatozoal measurements

M otility (%) 91.57 ± 0.73 89.70 ± 0.66 b 87.78 ± 1.10* 88.13 ± 0.56**

Sperm (10 6 /g cauda epididymis) 1,237 ± 60 1,204 ± 66 b 1,062 ± 70 890 ± 68**

Sperm (10 6 /cauda epididymis) 19.93 ± 0.47 19.66 ± 0.69 b 18.92 ± 0.97 16.93 ± 1.15

* Significantly different (P ≤ 0.0 5 ) from the chamber control group by Dunnett’s test (epididymis weights) or Shirley’s test (sperm motility)

** Significantly different (P ≤ 0.01) from the chamber control group by Shirley’s test

a Data are presented as mean ± standard error. Differences from the chamber control group are not significant by Dunnett’s test (body, cauda

epididymis, and testis weights) or Dunn’s test (spermatid m easurements and sperm/cauda epididymis ).

b n=9



### T ABLE 

### H4

Estrous Cycle Characterization for Female Mice in th e 3

- Month Inhalation Study of 1

- Bromopropane a



### Chamber Control 125 ppm 250 ppm 5 00 ppm

Number weighed at necropsy 10 10 10 5

Necropsy body wt (g) 30.9 ± 0.7 31.9 ± 0.8 30.2 ± 0.9 31.3 ± 1. 4

Proportion of regular cycling females b 9 /10 8 /10 6 /10 5 / 5

Estrous cycle length (days) 4. 2 ± 0. 11 4.6 ± 0. 50 4. 2 ± 0. 08 4.6 ± 0. 10*

Estrous stages (% of cycle) c

Diestrus 27.5 27.5 25.8 41.7

Proestrus 0.0 0 .0 0.0 0.0

Estrus 49.2 48.3 52.5 41.7

Metestrus 23.3 24.2 21.7 16.7

* Significantly different (P ≤ 0.0 5 ) from the chamber control group by Dunn’s test

a Necropsy body weights and estrous cycle length data are presented as mean ± standard error. Differences from the chamber control group are

not significant by Dunnett’s test (body weight).

b Number of females with a regular cycle/number of females cycling

c Evidence shows that females exposed to 500 ppm differ significantly (Wilk’s Criterion, P ≤ 0.05) from the chamber control females in the

relative length of time spent in the estrous stages. The tests for equality of transition probability matrices among exposed grou ps and between

the chamber control group and each exposed group indicated that 500 ppm females spent significantly more time in extended diestrus

(P=0.035) and 250 ppm females spent significantly more time in extended estrus (P<0.001) than the chamber cont rol group.

### 172 1- Bromopropane, NTP TR 564

### 173



# APPENDIX I

CHEMICAL CHARACTERIZ ATION AND

GENERATION OF CHAMBER CONCENTRATIONS

P ROCUREMENT AND 

### C HARACTERIZATION OF 

### 1-B ROMOPROPANE



### ........................................................................ 174

V APOR 

### G ENERATION AND 

### E XPOSURE 

### S YSTEM



### ....................................................................................................... 174

V APOR 

### C ONCENTRATION 

### M ONITORING



### .................................................................................................................. 175

C HAMBER 

### A TMOSPHERE 

### C HARACTERIZATION



### ...................................................................................................... 175

F IGURE 

### I1 Infrared Absorption Spectrum of 1- Bromopropane

....................................................................... 177

F IGURE 

### I2 1 

### H- Nuclear Magnetic Resonance Spectrum o f 1- Bromopropane

.................................................. 178

T ABLE 

### I1 Gas Chromatography Systems Used in the Inhalation Studies of 1- Bromopropane

................... 179

F IGURE 

### I3 Schematic of the Vapor Generation and Delivery System

in the Inhalation Studies of 1- Bromopropane

.................................................................................. 180

T ABLE 

### I2 Summary of Chamber Concentrations in the 2- Week Inhalation Studies

of 1- Bromopropane

............................................................................................................................ 181

T ABLE 

### I3 Summary of Chamber Concentrations in the 3- Month Inhalation Studies

of 1- Bromopropane

........................................................................................................................... 181

T ABLE 

### I4 Summary of Chamber Concentrations in the 2- Year Inhalation Studies

of 1- Bromopropane

........................................................................................................................... 182

### 174 1- Bromopropane, NTP TR 564



# CHEMICAL CHARACTERIZATION AND

GENERATION OF CHAMBER CONCENTRATIONS

P ROCUREMENT AND 

# C HARACTERIZATION OF 

# 1-B ROMOPROPANE

1- Bromopropane was obtained in 55

- gallon metal drums from Diaz Chemical Corporation ( Holley

, NY ) in on e lot

(106

- 015) and from Albemarle PCC (Thann, France) in one lot ( 1581313004 ). Lot 106

- 015 was used i n the 2

- week

and 3

- month studies, and lot 1581313004 was used during the 2

- year studie s. Identity and purity analyses were

conducted by the study laboratory a t Battelle Toxicology Northwest (Richland, WA), and by Chemir/Polytech

Laboratories, Inc. (Maryla nd Heights, MO). Reports on analyses performed in support of the 1- bromopropane

studies are on file at the National Institute of Environmental Health Sciences.

Both lots of the chemical, a colorless to pale yellow liquid with a strong

, characteristic odor, were identified as

1- bromopropane by Chemir/Polytech Laboratories, Inc., by infrared (IR) and 1 

### H- nuclear magnetic resonance (NMR)

spectroscopy. All spectra were consistent with the literature reference spectra ( Aldrich , 1993, 1997) o f

1- bromopropane

. Representative IR and 1 

### H- NMR spectra are presented in Figures I1 and I2 , respectively

.

The purity of each lot w as determined by the study laboratory using gas chromatography (GC) by system A

(Table I1). In addition, Chemir/Polytech L aboratories, Inc., determined the moisture content of each lot by Karl

Fischer titration and the purity of each lot by elemental and hydrogen bromide (HBr) analyses. The analyses of the

bulk chemical for HBr were performed by volumetric extraction of the samples with deionized water subsequent to

an analysis of the extractables for free bromide ion using ion chromatography (IC). IC was performed on a Dionex

DX100 ion chromatograph (Dionex Corporation, Sunnyvale, CA) equipped with a conductivity detector a nd IonPac

(Dionex) column

.

For lot 106

- 015 , Karl Fischer titration indicated a water content of 39 ppm

. E lemental analyses for carbon

, hydrogen , nitrogen, and sulfur were in agreement with the theoretical values for 1

- bromopropane; residual HBr was

determined to be 1.3 ppm. GC indicated one major peak and three impurities with areas exceeding 0.1% of the total

peak area; these peaks matched the retention times for prepared standards of 1

- propanol (0.14%), 2

- bromopropane

(0.11%), and di

- n

- propyl ether (0.74%). Measured purity of the bulk chemical was consistent throughout the

sampled metal drum, and t he overall purity of lot 106

- 015 was determined to be approximately 99 %.

For lot 1581313004 , Karl Fischer titration indicated a water content of 119 ppm . E lemental analyses for carbon

, hydrogen , nitrogen, and sulfur were in agreement with the theoretical values for 1

- bromopropane; residual HBr was

determined to be 61.6 ppm. GC detected one major peak and no impurities with areas greater than 0.1% of the total

peak area. Three impurities identified with prepared standards were 1

- propanol (0.03%), 2

- bromopropane (0.02%),

and di

- n

- propyl ether (0.02%). Measured purity of the bulk chemical was consistent throughout the sampled metal

drums, and t he o verall purity of lot 1581313004 was determined to be 99. 9% or greater

.

To ensure stability, t he bulk chemical was stored at controlled room temperature in the original shipping containers

( 55

- gallon metal drums )

. Periodic reanalyses of the bulk chemical were performed during the 2

- week, 3

- month, and

2- year studies by the study laboratory using GC by system A

, and n o degradation of the bulk chemical was detected.



# V APOR 

# G ENERATION AND 

# E XPOSURE 

# S YSTEM

A diagram of the 1- bromopropane vapor generation and delivery system used in the studies is shown in Figure I3.

1- Bromopropane was pumped through a preheater ( for the 2

- week and 3

- month studies ) and into a heated glass

column filled with glass beads that increased the surface area for vaporization. Heated nitrogen entered the column

from below and assisted in vaporizing the chemical while conveying it into a short distribution manifold

. Concentration in the manifold was determined by the chemical pump rate and nitrogen flow rate. The pressure in

the distribution manifold was kept fixed to ensure constant flow through the manifold and into all chambers as the

flow of vapor to each chamber was adjusted.

### 1- Bromopropane, NTP TR 564 175

M etering valve s at the manifold controlled flow to each chamber through individual T eflon ® 

### delivery lines that

carried the vapor from the manifold to three

- way exposure valves at the chamber inlets. The exposure valves

diverted vapor delivery to exposure chamber exhaust until the generation system was stable and exposures were

ready to proceed. To initiate exposure , the chamber exposure valves were rotated to allow the 1

- bromopropane

vapor to flow to each exposure chamber inlet duct where it was further diluted with filtered, conditioned air to

achieve the desired exposure concentration.

The study laboratory designed the inhalation exposure chamber (Harford Systems Division of Lab Products, Inc.,

Aberdeen, MD) so that uniform vapor concentrations could be maintained throughout the chamber with the catch

pans in place. The total active m ixing volume of each chamber was 1.7 m 3 

### . A small particle detector

(Model 3022A, TSI Incorporated, St. Paul, MN) was used with and without animals in the exposure chambers to

ensure that 1- bromopropane vapor, and not aerosol, was produced. No particle co unts above the minimum

resolvable level (approximately 200 particles/cm 3 

### ) were detected.



# V APOR 

# C ONCENTRATION 

# M ONITORING

Summaries of the chamber vapor concentrations are given in Tables I2 through I 4. Chamber and room

concentrations of 1

- bromopropane were monitored by an on

- line gas chromatograph (system B, Table I1). Samples

were drawn from each exposure chamber approximately every 20 (2

- week and 3

- month studies) or 30 (2

- year

studies) minutes during each 6

- hour exposure period using Hastelloy ® 

### -C str eam

- select and gas

- sampling valves

(VALCO Instruments Company, Houston, TX) in a separate, heated valve oven. The sample lines composing each

sample loop were made from Teflon ® 

### tubing and were connected to the exposure chamber relative humidity

sampling l ines at a location close to the gas chromatograph. A vacuum regulator maintained a constant vacuum in

the sample loop to compensate for variations in sample line pressure. An in

- line flow meter between the vacuum

regulator and the gas chromatograph allow ed digital measurement of sample flow.

The on

- line gas chromatograph was checked throughout the day for instrument drift against an on

- line standard

vapor of 1

- bromopropane in nitrogen supplied by a standard generator (Kin

- Tek ; Precision Calibration Syste ms,

La Marque, TX). The on

- line gas chromatograph was re calibrated as required to meet acceptance criteria.

Calibration was performed by a comparison of chamber concentration data to data from grab samples that were

collected with activated coconut charc oal gas sampling tubes (ORBO TM- 32 ; Supelco, Bellefonte, PA), extracted with

methylene chloride containing 1

- bromobutane as an internal standard, and analyzed using an off

- line gas

chromatograph (system C ). Known volumes of chamber atmosphere were sampled a t a constant flow rate ensured

by a calibrated critical orifice. The off

- line gas chromatograph was calibrated with gravimetrically prepared

standards of 1

- bromopropane containing 1

- bromobutane as an internal standard in methylene chloride.



# C HAMBER 

# A TMOSPHERE 

# C HARACTERIZATION

Buildup and decay rates for chamber vapor concentrations were determined with and without animals present in the

chambers. At a chamber airflow rate of 15 air changes per hour, the theoretical value for the time to achieve 90% of

t he target concentration after the beginning of vapor generation (T 90 

### ) and the time for the chamber concentration to

decay to 10% of the target concentration after vapor generation was terminated (T 10 

### ) was approximately

12.5 minutes. For rats and mice in the 2- week s tudies , T 90 

### and T 10 

### values ranged from 9 to 10 minutes with animals

present . For rats and mice in the 3- month studies, T 90 

### values averaged 9 minutes without animals present and

ranged from 9 to 10 minutes with animals present; T 10 

### values r anged from 8 to 9 minutes without animals present

and from 9 to 11 minutes with animals present. For rats and mice in the 2

- year studies , T 90 

### values ranged from 9 to

10 minutes without animals present and from 9 to 11 minutes with animals present ; T 10 

### val ues ranged from 9 to

10 minutes without animals present and from 10 to 11 minutes w ith animals present . A T 90 

### value of 12 minutes was

selected for the 2

- week studies, and a T 90 

### value of 10 minutes was selected for the 3

- month and 2

- year studies.

The unif ormity of vapor concentration in the inhalation exposure chambers without animals present was evaluated

before the 3- month and 2

- year studies began ; in addition, concentration uniformity with animals present in the

chambers was measured once during the 2- week studies

, once during the 3- month studies , and quarterly during the

2- year studies. The vapor concentration was measured using the on

- line gas chromatograph (system B , Table I1)

with the stream

- selection valve fixed in one position to allow continuous monitoring from a single input line.

### 176 1- Bromopropane, NTP TR 564

During the 2

- week studies and prior to the 3

- month and 2

- year studies, concentrations were measured at 12 chamber

positions, one in front and one in back for each of the six possible animal cage unit positions per c hamber. During

the 3

- month and 2

- year studies, concentrations were measured at the regular monitoring port and from sample ports

at levels where animals were present. Chamber concentration uniformity was maintained throughout the studies.

The persistenc e of 1

- bromopropane in the chambers after vapor delivery ended was determined by monitoring the

vapor concentration in the 2,000 ppm chamber s in the 2- week studies, the 500 ppm (rats and mice) and 1,000 ppm

(rats) chambers in the 3- month studies, and the 2 50 ppm (mice) and 500 ppm (rats) chambers in the 2

- year s tudies

, with (all studies) and without (3

- month and 2

- year studies) animals present in the chambers. In the 2- week studies

, the concentration decreased to 1% of the target concentration within 19 mi nutes with animals present. In the

3- month studies, the concentration decreased to 1% of the target concentration within 20 minutes without animals

present and within 25 ( rat and mouse chamber s ) and 21 ( rat chamber) minutes with animals present. In the 2- year

studies, the concentration decreased to 1% of the target concentration within 19 (rats) and 20 (mice) minutes without

animals present and within 25 (rats) and 22 (mice) minutes with animals present

.

Samples of the test atmosphere from the distribut ion lines and low and high exposure concentration chambers were

collected prior to the 3

- month and 2

- year studies and also at the beginning and end of one generation day during the

2- week, 3

- month, and 2

- year studies; the atmosphere samples were collected with adsorbent gas sampling tubes

containing activated charcoal (ORBO TM- 32), followed by a tube containing silica gel (ORBO TM- 52; Supelco, Inc.)

and extracted with methylene chloride. Additional samples were collected from the generator reservoir, and all o f

the samples were analyzed using GC by system A to measure the stability and purity of 1

- bromopropane in the

generation and delivery system. To assess whether impurities or degradation products co

- eluted with

1- bromopropane or the solvent, a second GC an alysis of the samples was performed using a polar column capable of

resolving compounds with similar boiling points and polarities (system D ). Fourier transform IR was used to

determine if HBr was present in the atmosphere samples ; spectra were generated using a MIDAC I

- 1101

spectrometer with a 9.5 m path

- length gas cell (MIDAC Corporation, Irvine, CA) and were compared to those of

prepared HBr standards.

No evidence of degradation of 1

- bromopropane was noted in any part of the exposure system. Three impurity peaks

with areas greater than 0.1% of the total peak area were consistently detected in atmosphere and generator reservoir

samples collected prior to the 3

- month studies and during the 2

- week and 3

- month studies. These peaks matched the

retention times for prepared standards of 1

- propanol, 2

- bromopropane, and di

- n

- propyl ether and had area percent

values similar to those measured during the initial bulk purity analyses of lot 106

- 015. No impurity peaks were

detected in atmosphere or generator reservoir samples collected pr i or to or during the 2

- year studies. Using the

polar column, no additional impurities were detected in any of the atmosphere or generator reservoir samples

collected prior to or during any of the studies. HBr levels in the expo sure atmosphere were determined to be less

than 0.07% during the 2

- week studies and less than 1% prior to and during the 3

- month and 2

- year studies.

### 1- Bromopropane, NTP TR 564 177

F IGURE 

### I1

Infrared Absorption Spectrum of 1

- Bromopropane

### 178 1- Bromopropane, NTP TR 564

F IGURE 

### I2

1 

### H- Nuclear Magnetic Resonance Spectrum of 1

- Bromopropane

### 1- Bromopropane, NTP TR 564 179

T ABLE 

### I1

Gas Chromatography Systems Used in the Inhalation Studies of 1

- Bromopropane a



### Detection System Column Carrier Gas Oven Temperature

Program

System A

Flame ionization DB

- 5, 30 m × 0.53 mm,

5.0

- μm film thickness

(J&W Scientific , Folsom, CA) or

Rtx

- 5, 30 m × 0.53 mm,

5.0

- μm film thickness ( Restek

, Bell e fonte, PA ) Helium at 4 psi head pressure 35° C for 3 minutes, then

3° C/minute to 65° C, then

7° C/minute to 260° C, held for

1 minute



### System B

Flame ionization DB

- 5, 15 m × 0.53 mm,

1.5

- μm film thickness

(J&W Scientific ) Nitrogen at 17 mL/minute Isothermal at 35° C



### System C

Flame ionization DB

- 5, 30 m × 0.53 mm,

5.0

- μm film thickness

(J&W Scientific ) or

Rtx

- 5, 30 m × 0.53 mm,

5.0

- μm film thickness ( Restek) Helium at 6 psi head pressure 40 ° C for 1 minute, then

6 ° C/minute to 9 5° C, then

15 ° C/minute to 15 0° C, held for

2 minute s



### System D

Flame ionization DB

- Wax , 30 m × 0.53 mm,

1 .0

- μm film thickness

(J&W Scientific ) Helium 40 ° C for 1 minute, then

6 ° C/minute to 9 5° C, then

15 ° C/minute to 15 0° C, held for

2 minute s

a The gas chromatographs were manufactured by Hewlett

- Packard (Palo Alto, CA).

### 180 1- Bromopropane, NTP TR 564

F IGURE 

### I3

Schematic of the Vapor Generation and Delivery System in the Inhalation Studies

of 1

- Bromopropane

### 1- Bromopropane, NTP TR 564 181

T ABLE 

### I2

Summary of Chamber Concentrations in the 2

- Week Inhalation Studies of 1- Bromopropane

Target Concentration

(ppm) Total Number of Readings Average Concentration a



### (ppm)

Rat Chambers

125 225 127 ± 2

250 226 252 ± 3

500 227 499 ± 4

1,000 226 1,015 ± 12

2,000 228 2,028 ± 17



### Mouse Chambers

125 245 127 ± 2

250 246 251 ± 3

500 247 499 ± 3

1,000 246 1,013 ± 13

2,000 248 2,027 ± 17

a Mean ± standard deviation



### T ABLE 

### I3

Summary of Chamber Concentrations in the 3

- Month Inhalation Studies of 1- Bromopropane

Target Concentration

(ppm) Total Number of Readings Average Concentration a



### (ppm)

Rat Chambers

62.5 1,222 62.2 ± 1.2

1 25 1,237 124 ± 2

250 1,247 247 ± 4

500 1,241 497 ± 8

1,000 1,243 1,005 ± 15



### Mouse Chambers

6 2

. 5 1,264 62.1 ± 1.2

1 25 1,277 124 ± 2

2 50 1,287 247 ± 4

5 00 1,281 497 ± 8

a Mean ± standard deviation

### 182 1- Bromopropane, NTP TR 564

T ABLE 

### I4

Summary of Chamber Concentrations in the 2

- Year Inhalation Studies of 1- Bromopropane

Target Concentration

(ppm) Total Number of Readings Average Concentration a



### (ppm)

Rat Chambers

125 8,018 125 ± 3

250 8,011 250 ± 6

500 8,042 500 ± 11



### Mouse Chambers

62.5 8,494 62.5 ± 1.6

125 8,009 125 ± 3

250 8,038 250 ± 5

a Mean ± standard deviation

### 183



# APPENDIX J

INGREDIENTS, NUTRIENT COMPOSITION,

AND CONTAMINANT LEVE LS

IN NTP

- 2000 RAT AND MOUSE R ATION

T ABLE 

### J1 Ingredients of NTP

- 2000 Rat and Mouse Ration

............................................................................ 184

T ABLE 

### J2 Vitamins and Minerals in NTP

- 2000 Rat and Mouse Ration

......................................................... 184

T ABLE 

### J3 Nutrient Composition of NTP

- 2000 Rat and Mouse Ration

........................................................... 185

T ABLE 

### J4 Contaminant Levels in NTP

- 2000 Rat and Mouse Ration

............................................................. 186

### 184 1- Bromopropane, NTP TR 564

T ABLE 

### J1

Ingredients of NTP

- 2000 Rat and Mouse Ration

Ingredients Percent by Weight

Ground hard winter wheat 22.26

Ground #2 yellow shelled corn 22.18

Wheat middlings 15.0

Oat hulls 8.5

Alfalfa meal (dehydrated, 17% protein) 7.5

Purified cellulose 5.5

Soybean meal (49% protein) 5.0

Fish meal (60% protein) 4.0

Corn oil (without preservatives) 3.0

Soy oil (without preservatives) 3.0

Dried brewer’s yeast 1.0

Calcium carbonate (USP) 0.9

Vitamin premix a 0.5

Mineral premix b 0.5

Calcium phosphate, dibasic (USP) 0.4

Sodium chloride 0.3

Choline chloride (70% choline) 0.26

Methionine 0.2

a Wheat middlings as carrier

b Calcium carbonate as carrier



### T ABLE 

### J2

Vitamins and Minerals in NTP

- 2000 Rat and Mouse Ration a



### Amount Source

Vitamins

A 4,000 IU Stabilized vitamin A palmitate or acetate

D 1,000 IU D

- activated animal sterol

K 1.0 mg Menadione sodium bisulfite complex

α

- Tocopheryl acetate 100 IU

Niacin 23 mg

Folic acid 1.1 mg

d

- Pantothenic acid 10 mg d

- Calcium pantothenate

Riboflavin 3.3 mg

Thiamine 4 mg Thiamine mononitrate

B 12 52 μg

Pyridoxine 6.3 mg Pyridoxine hydrochloride

Biotin 0.2 mg d

- Biotin



### Minerals

Magnesium 514 mg Magnesium oxide

Iron 35 mg Iron sulfate

Zinc 12 mg Zinc oxide

Manganese 10 mg Manganese oxide

Copper 2.0 mg Copper sulfate

Iodine 0.2 mg Calcium iodate

Chromium 0.2 mg Chromium acetate

a Per kg of finished product

### 1- Bromopropane, NTP TR 564 185

T ABLE 

### J3

Nutrient Composition of NTP

- 2000 Rat and Mouse Ration

Nutrient Mean ± Standard

Deviation Range Number of Samples

Protein (% by weight) 1 4.8 ± 0. 65 13 .7 – 16.1 24

Crude fat (% by weight) 8.1 ± 0. 36 7. 4

– 9.0 2 4

Crude fiber (% by weight) 9. 2 ± 0. 45 8.2

– 9.9 2 4

Ash (% by weight) 5.0 ± 0. 25 4. 4

– 5.4 2 4



### Amino Acids (% of total diet)

Arginine 0.7 70 ± 0.0 7 0 0.670

– 0. 97 0 1 8

Cystine 0.22 5 ± 0.02 3 0. 15 0

– 0.2 5 0 1 8

Glycine 0. 706 ± 0.04 3 0.620

– 0. 80 0 1 8

Histidine 0.3 62 ± 0.0 82 0.310

– 0. 68 0 1 8

Isoleucine 0.524 ± 0.046 0.430

– 0. 66 0 1 8

Leucine 1.0 87 ± 0.06 6 0.960

– 1. 24 0 1 8

Lysine 0.7 12 ± 0. 118 0. 310

– 0. 84 0 1 8

Methionine 0.40 7 ± 0.05 1 0. 26 0

– 0.4 9 0 1 8

Phenylalanine 0. 626 ± 0.0 4 3 0.540

– 0. 72 0 1 8

Threonine 0.50 0 ± 0.0 46 0.430

– 0. 61 0 18

Tryptophan 0.1 4 2 ± 0.0 2 4 0.110

– 0. 20 0 1 8

Tyrosine 0.3 88 ± 0.05 8 0.280

– 0. 5 40 1 8

Valine 0.6 67 ± 0.04 5 0.550

– 0. 73 0 1 8



### Essential Fatty Acids (% of total diet)

Linoleic 3.9 2 ± 0.24 3 3. 4 9

– 4.54 1 8

Linolenic 0.30 ± 0.0 35 0.21

– 0.35 1 8



### Vitamins

Vitamin A (IU/kg) 4

, 870 ± 121 3, 230

– 8 ,9 0 0 2 4

Vitamin D (IU/kg) 1,000 a

α

- Tocopherol (ppm) 8 4 .2 ± 1 6

. 60 52.0

– 1 10 .0 15

Thiamine (ppm) b 9.0 ± 3.71 6. 4 – 25.2 24

Riboflavin (ppm) 6 .8 ± 2.11 4.20

– 11

. 2 0 15

Niacin (ppm) 79.0 ± 10.50 66.4

– 98.2 15

Pantothenic acid (ppm) 23.9 ± 3. 73 17.4

– 29. 8 15

Pyridoxine (ppm) b 9. 21 ± 2. 20 6.4

– 1 3.7 15

Folic acid (ppm) 1. 75 ± 0. 54 1.2 0

– 3 .2 7 15

Biotin (ppm) 0.33 2 ± 0.1 2 0.225

– 0.704 15

Vitamin B 12 (ppb) 6 0.5 ± 46.5 18.3

– 174.0 15

Choline (ppm) b 3, 064 ± 2 70 2,700

– 3,790 15



### Minerals

Calcium (%) 0.9 64 ± 0.043 0. 884

– 1.0 3 0 2 4

Phosphorus (%) 0.5 7 8 ± 0.02 6 0. 535

– 0. 623 2 4

Potassium (%) 0. 665 ± 0.02 3 0.62 6

– 0.69 4 15

Chloride (%) 0.37 6 ± 0.0 41 0.300

– 0. 474 15

Sodium (%) 0.19 1 ± 0.01 7 0.160

– 0.2 2 2 15

Magnesium (%) 0. 201 ± 0.009 0.185

– 0.21 7 15

Sulfur (%) 0.17 0 ± 0.02 9 0.1 16

– 0.209 15

Iron (ppm) 1 82 ± 46 .7 135

– 311 15

Manganese (ppm) 5 4.1 ± 7.89 42.1

– 73.1 15

Zinc (ppm) 5 5

. 0 ± 9.55 43.3

– 78.5 15

Copper (ppm) 6. 65 ± 1.790 3.21

– 10.50 15

Iodine (ppm) 0. 512 ± 0. 221 0.233

– 0. 972 15

Chromium (ppm) 0. 604 ± 0.2 53 0.330

– 1

. 380 14

Cobalt (ppm) 0.2 5 ± 0.0 74 0.20

– 0. 47 14

a From formulation

b As hydrochloride (thiamine and pyridoxine) or chloride (choline)

### 186 1- Bromopropane, NTP TR 564

T ABLE 

### J4

Contaminant Levels in NTP

- 2000 Rat and Mouse Ration a



### Mean ± Standard

Deviation b 

### Range Number of Samples

Contaminants

Arsenic (ppm) 0. 25 ± 0. 120 0.1 4

– 0.50 2 4

Cadmium (ppm) 0.0 7 ± 0.0 22 0.04

– 0. 10 2 4

Lead (ppm) 0.09 ± 0.0 23 0.05

– 0.13 24

Mercury (ppm) <0.02 2 4

Selenium (ppm) 0. 21 ± 0.0 58 0.1 6

– 0. 45 2 4

Aflatoxins (ppb) <5.00 2 4

Nitrate nitrogen (ppm) c 13.1 ± 4.17 7.89

– 24.4 24

Nitrite nitrogen (ppm) c <0.61 2 4

BHA (ppm) d <1.0 24

BHT (ppm) d <1.0 24

Aerobic plate count (CFU/g) 10 ± 0.0 10 2 4

Coliform (MPN/g) 3.0 ± 0.0 3.0 2 4

Escherichia coli (MPN/g) <10 2 4

Salmonella (MPN/g) Negative 2 4

Total nitrosoamines (ppb) e 4. 4 ± 1.7 3 2. 3 – 8. 5 24

N

- Nitrosodimethylamine (ppb) e 2.4 ± 1.29 1. 1

– 5. 6 2 4

N

- Nitrosopyrrolidine (ppb) e 1.9 ± 0.80 0 .9

– 4.1 2 4



### Pesticides (ppm)

α

- BHC <0.01 2 4

β

- BHC <0.02 2 4

γ

- BHC <0.01 2 4

δ

- BHC <0.01 2 4

Heptachlor <0.01 2 4

Aldrin <0.01 2 4

Heptachlor epoxide <0.01 2 4

DDE <0.01 2 4

DDD <0.01 2 4

DDT <0.01 2 4

HCB <0.01 2 4

Mirex <0.01 2 4

Methoxychlor <0.05 24

Dieldrin <0.01 2 4

Endrin <0.01 2 4

Telodrin <0.01 2 4

Chlordane <0.05 2 4

Toxaphene <0.10 2 4

Estimated PCBs <0.20 2 4

Ronnel <0.01 2 4

Ethion <0.02 2 4

Trithion <0.05 2 4

Diazinon <0.10 2 4

Methyl chlorpyrifos 0.1 22 ± 0.1 39 0.020

– 0.4 16 2 4

Methyl parathion <0.02 2 4

Ethyl parathion <0.02 2 4

Malathion 0. 165 ± 0.1 78 0.020

– 0.589 2 4

Endosulfan I <0.01 2 4

Endosulfan II <0.01 2 4

Endosulfan sulfate <0.03 2 4

a All samples were irradiated. CFU=colony

- forming units; MPN=most probable number; BHC=hexachlorocyclohexane or

benzene hexachloride

b For values less than the limit of detection, the detection limit is given as the mean.

c Sources of contamination: alfalfa, grains, and fish meal

d Sources of contamination: soy oil and fish meal

e All values were corrected for percent recovery.

### 187



# APPENDIX K

SENTINEL ANIMAL PROG RAM

Methods

................................................................................................ .................................................................... 188

Results

................................................................................................ ....................................................................... 190

### 188 1- Bromopropane, NTP TR 564



# SENTINEL ANIMAL PROG RAM

M ETHODS

Rodents used in the Carcinogenesis Program of the National Toxicology Program are produced in optimally clean

facilities to eliminate potential pathogens that may affect study results. The Sentinel Animal Program is part of the

periodic monitoring of animal health that occurs during the toxicologic evaluation of chemical compounds. Under

this program, the disease state of the rodents is monitored via sero logy on sera from extra (sentinel) animals in the

study rooms. These animals and the study animals are subject to identical environmental conditions. The sentinel

animals come from the same production source and weanling groups as the animals used for the studies of chemical

compounds.

Serum samples were collected from five male and five female chamber control rats and mice at the end of the

2- week and 3

- month studies, five male and five female sentinel rats and mice 1 week after the start of the 3

- month

and 2

- year studies, five male and five female sentinel rats and mice at 6 and 12 months in the 2

- year studies, five

male and four female sentinel rats at 18 months in the 2

- year studies, five male and five female mice at 18 months in

the 2

- year studies, and five males and five females from the 500 ppm rats and 250 ppm mice at the end of the 2

- year

studies. Blood from each animal was collected and allowed to clot, and the serum was separated ; fecal samples

were collected from five male and five female mice at 18 months in the 2

- year study for Helicobacter species by

polymerase chain reaction testing. The samples were processed appropriately and sent to MA Bioservices/

BioReliance (Rockville, MD) for determination of antibody titers. The laboratory methods and agents for which

testing was performed are tabulated below; the times at which samples were collected during the studies are also

listed.

Method and Test



# R ATS Time of Collection

2

- Week Study

ELISA

H

- 1 (Toolan’s H

- 1 virus) Study termination

KRV (Kilham rat virus) Study termination

Mycoplasma pulmonis Study termination

PVM (pneumonia virus of mice) S tudy termination

RCV /SDA Study termination

(rat coronavirus /sialodacryoadenitis virus )

Sendai S tudy termination

3

- Month Study

ELISA

H

- 1 1 week

KRV 1 week

Mycoplasma arthritidis Study termination

M. pulmonis 1 week, study termination

PVM 1 week, study termination

RCV/SDA 1 week, study termination

Sendai 1 week, study termination

Immunofluorescence Assay

Parvovirus Study termination

### 1- Bromopropane, NTP TR 564 189

Method and Test



# R ATS (continued) 

### Time of Collection

2

- Year Study

ELISA

H

- 1 1 week

KRV 1 week

M

. pulmonis 1 week, study termination

M. arthritidis Study termination

PVM 1 week, 6, 12, and 18 months, study termination

RCV/SDA 1 week, 6, 12, and 18 months, study termination

Sendai 1 week, 6, 12, and 18 months, study termination

Immunofluorescence Assay

M. arthritidis Study termination

PVM 18 months

RCV/SDA 12 and 18 months

Sendai 18 months

Parvovirus 6, 12, and 18 months, study termination



# M ICE

2

- Week Study

ELISA

GDVII (mouse encephalomyelitis virus) Study termination

MVM (minute virus of mice) Study termination

MHV (mouse hepatitis virus) Study termination

M. pulmonis Study termination

PVM S tudy termination

Sendai Study termination

3

- Month Study

ELISA

Ectromelia virus Study termination

EDIM (epizootic diarrhea of infant mice) Study termination

GDVII 1 week, study termination

LCM (lymphocytic choriomeningitis virus) Study termination

MVM 1 week

Mouse adenoma virus

- FL Study termination

MHV 1 week, study termination

M. arthritidis Study termination

M. pulmonis 1 week, study termination

PVM 1 week, study termination

Reovirus 3 Study termination

Sendai 1 week, study termination

Immunofluorescence Assay

MCMV (mouse cytomegalovirus) Study termination

Parvovirus Study termination

### 190 1- Bromopropane, NTP TR 564

Method and Test



# M ICE (continued) 

### Time of Collection

2

- Year Study

ELISA

Ectromelia virus 6, 12, and 18 months, study termination

EDIM 6, 12, and 18 months, study termination

GDVII 1 week, 6, 12, and 18 months, study termination

LCM 6, 12, and 18 months, study termination

MVM 1 week, 12 and 18 months, study termination

Mouse adenoma virus 6, 12, and 18 months, study termination

MHV 1 week, 6, 12, and 18 months, study termination

M. arthritidis Study termination

M. pulmonis 1 week, study termination

PVM 1 week, 6, 12, and 18 months, study termination

Reovirus 3 6, 12, and 18 months, study termination

Sendai 1 week, 6, 12, and 18 months, study termination

Immunofluorescence Assay

Ectromelia virus 12 months, study termination

EDIM 12 months

GDVII 12 months

Mouse adenoma virus

- FL 18 months

MCMV Study termination

MHV Study termination

M. pulmonis Study termination

Reovirus 3 12 months

Parvovirus 6 and 18 months, study termination

Polymerase Chain Reaction

Helicobacter species 18 months



# R ESULTS

All test results were negative.