## 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)

Scheduled Peer Review Date: November 19 , 200 9

NOTICE

This DRAFT Technical Report is distributed solely for the purpose of predissemination peer review under the applicable inform ation

quality guidelines. It has not been formally disseminated by the NTP. It does not represent and should not be construed to represent

NTP determination or policy

.



## NTP TR 564

NIH Publication No. 10

- 5906



# National Toxicology Program

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 Nationa l Institute for Occupational 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 c oordinating 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, scienti fic and

medical communities, 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 Techni cal Report series are

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

a nd carcinogenicity studies 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 studie s in compliance with its laboratory health and safety guidelines and FDA Good

Laboratory 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 Heal th 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 char ge

electronically 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)

Scheduled Peer Review Date: November 19 , 200 9

NOTICE

This DRAFT Technical Report is distributed solely for the purpose of predissemination peer review under the applicable information

quality guidelines. It has not been formally disseminated by the NTP. It does not represent and should not be construed to represent

NTP determination or policy.



## NTP TR 564

NIH Publication No. 10

- 5906



# National Toxicology Program

National Institutes of Health

Public Health Service

U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES

### 2

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# 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 evaluated 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

. Wan g, 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



### P. M. Foster , Ph.D.

, Observer

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.



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

, Observer

National Toxicology Program



### S. Newbigging, M.Sc., D.V.M., D.V.Sc., Observer

ILS, Inc.



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

.

National Toxicology Program



### R.E. Sills, D.V.M., Ph.D.

, Observer

National Toxicology Program



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

National Toxicology Program

### 1- Bromopropane , NTP TR 564 3

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

## 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



### P. M. Foster , Ph.D., Observer

National Toxicology Program



### S.L. Grumbein

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

Battelle Toxicology Northwest



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

GlaxoSmithKline



### T. Kojitani, D.V.M., Ph.D., Observer

National Toxicology Program



### 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.



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

National Toxicology Program



### 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 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft

### 5

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# C ONTENTS

ABSTRACT

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

EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY

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

TECHNICAL REPORTS REVIEW SUBCOMMITTEE

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

SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMITTEE COMMENTS

................................ 16

INTRODUCTION

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

MATERIALS AND METHODS

.............................................................................................................................. 31

RESULTS

................................................................................................ ................................................................... 51

DISCUSSION AND CONCLUSIONS

..................................................................................................................... 97

REFERENCES

................................................................................................ ........................................................ 10 5

A PPENDIX 

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

- Year Inhalation Study

of 1- Bromopropane

..................................................................................................................... A-1

A PPENDIX 

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

- Year Inhalation Study

of 1- Bromopropane

.................................................................................................................... B-1

A PPENDIX 

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

- Year Inhalation Study

of 1- Bromopropane

.................................................................................................................... C-1

A PPENDIX 

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

- Year Inhalation Study

of 1- Bromopropane

.................................................................................................................... D-1

A PPENDIX 

### E Genetic Toxicology

...................................................................................................................... E-1

A PPENDIX 

### F Clinical Pathology Results

.......................................................................................................... F-1

A PPENDIX 

### G Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios

.................................................. G-1

A PPENDIX 

### H Reproductive Tissue Evaluations and Estrous Cycle Characterization

................................ H-1

### 6 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft A PPENDIX 

### I Chemical Characterization and

Generation of Chamber Concentrations

.................................................................................... I-1

A PPENDIX 

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

in NTP

- 2000 Rat and Mouse Ration

........................................................................................... J-1

A PPENDIX 

### K Sentinel Animal Program

.......................................................................................................... K-1

### 7

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# ABSTRACT

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 pesticides,

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

e missive 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 afety and H ealth A dministration based on the potential for widespread

occupational and environmental exposure and a lack of toxicity and carcinogenicity 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.

### 8 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 

# 2-W EEK 

# S TUDY IN 

# R ATS

Groups of five male and five female rats were exposed to 1

- bromopropane vapor at concentrations o f 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 significantly 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 expos ed groups of females were

significantly increased. 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

incr eased

. Nasal lesions included s uppurative 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

Group s 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 1,000 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

. Kidney

weights of 1,000 and 2,000 ppm females were significantly increased. Microscopic lesions relate d to

1- bromopropane exposure occurred in 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 concen trations 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

### 1- Bromopropane , NTP TR 564 9

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION surviv ed 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 hepatotoxicity 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 kidney weights of 1,000 ppm females were significantly increased. Results of sperm count

and vaginal cytology evaluations showed exposure concentration

- related decreases in sperm motility and counts in

male rats, reaching 2 8 % and 37%, respectively, in the 1,000 ppm group. Female rats in all thr ee expos ure groups

evaluated exhibited altered estrous cycle s, spending significantly 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

observed 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 similar to

those of the chamber contr ols. 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 we ights of 500 ppm males were significantly

decreased. Male mice in the 500 ppm group had decreased sperm counts that 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 significantly 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.

### 10 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 

# 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 mac roscopic, soft, pale

- yellow to green, variably sized nodules were seen predominantly 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 s hown to be suppurative inflammation , many with

Splendore

- Hoeppli material.

The incidence of adenoma of the large intestine (colon or rectum ) in 500 ppm females was significantly greater than

that 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.

T he incidences of keratoacanthoma, basal cell adenoma, basal cell carcinoma, or squamous cell carcinoma

(combined) in all exposed groups of males were significantly greater than that in the chamber control group and

exceeded the historical control range for inhalation studies. The incidences of keratoacanthoma and of

keratoacanthoma or squamous cell carcinoma (combined) in 250 and 500 ppm mal es were also significantly

increased and exceeded the historical control range s for inhalation studies. In 500 ppm females, the incidence of

squamous cell papilloma, keratoacanthoma, basal cell adenoma, or basal cell carcinoma (combined) exceeded the

hist orical control range for inhalation studies.

The incidence of malignant mesothelioma in 500 ppm males was significantly greater than that in the chamber

control group.

The incidences of pancreatic islet adenoma in all exposed groups of males and of pancreatic islet adenoma or

carcinoma (combined) in 125 and 250 ppm males were significantly increased.

### 1- Bromopropane , NTP TR 564 11

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION Treatment

- related nonneoplastic lesions were observed in the respiratory system of exposed male and female rats.

In the nose, the incidences of suppurative chronic inflammation, chronic active inflammation, 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

metaplasia (except 125 ppm females) were increased in all exposed groups, and the incidences of suppurative

chronic inflammation were increased in the 500 ppm groups. In the trachea, there were increased incidence s 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 alveolar/bronchiolar adenoma, alveolar/bronchiolar carcinoma,

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 significantly

increased incidences of cytoplasmic vacuolization of the bronchiolar epithelium in all exposed male groups and

regeneration of the bronchiolar epithelium in all exposed groups of males and females

.

In the nose, there were significantly increased incidences 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 hyperplasia in all exposed female groups and in 62.5 and 250 ppm males. There were

significantly increased incidences of respiratory metaplasia of olfactory epithelium in 62.5 and 125 ppm m ales and

125 and 250 ppm females.

### 12 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T here were significantly increased incidences of cytoplasmic 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 conducted with

and without induced rat liver activation enzymes. Bacterial strains tested included S. typhimurium strains TA97,

TA98, TA100, and TA1535, as w ell as E. coli strain WP2 uvrA /pKM101. In addition, no increases in the

frequencies of micronucleated 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

Un der 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 neoplasms of the skin. Increased incidences of malignant mesothelioma and pancreatic islet adenoma

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 incidences of adenoma of the large intestine. Increased

incidences of neoplasms of the skin may also have been related to 1- bromopropane 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 carcinogenic activity of 1

- bromopropane in femal e

B6C3F1 mice based on increased incidences of alveolar/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, the trachea of female rats and male and female mice, and the lung of 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 14

.

### 1- Bromopropane , NTP TR 564 13

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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,

m etaplasia, 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)

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

epithel ium, hyperplasia

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

26/50) ; olfactory

epithelium, metaplasia,

respiratory (0/50, 7/50,

6/50, 3/50)

Larynx : vacuolization

cytoplasmic (0/48, 5/50,

10/48, 11/50)

Trachea : vacuolization

cytoplasmic (0/49, 15/50,

24/47, 24/50) Lung : bro nchiole,

regeneration ( 0/50, 45/50,

43/50, 49/50)

Nose : respiratory

epithelium, vacuolization

cytoplasmic (0/50, 3/50,

5/50, 8/50); respiratory

epithelium, hyperplasia

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

27/50); olfactory

epithelium, metaplasia,

respiratory (0/50, 4/5 0,

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 : adenoma

(0/50, 1/50, 2/50, 5/50) None Lung :

alveolar/bronchiolar

adenoma (1/50, 6/50,

4/50, 10/50);

alveola r/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) Skin : squamous cell

papilloma,

keratoacanthoma, basal

cell adenoma, or basal

cell carcinoma (1/50,

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

Level of evidence of

carcinogenic activity Some evidence Clear evidence No evidence Clear evidence

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

### 14 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 

## 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 de monstrate that a chemical is carcinogenic for laboratory animals under the conditions of the study and indicate that exposure to

the chemical has the potential for hazard to humans. Other organizations, such as the International Agency for Research on C an cer, 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 determination 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) increase of a combination of malignant and benign neoplasms, or (iii) marked increase of benign neo plasms

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 (mal ignant, benign, or combined) in which the strength of the response is less than that required for clear

evidence.

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

neoplasms that m ay 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 carcinogenic activity.

For studies showing multiple chemical

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

categories, the following convention has been adopted to convey completely the study results. 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 ex tend the actual

boundary of an individual category of evidence. Such consideration should allow for incorpora tion of scientific experience and current

understanding of long

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

two adjacent levels. These considerations should include:

**•** adequacy o f the experimental design and conduct;

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

**•** progression (or lack thereof) from benign to malignant neoplasia 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 pot ential to become malignant;

**•** combining benign and malignant tumor incidence known or thought to represent stages of progression in the same organ or tissu e;

**•** latency in tumor induction;

**•** multiplicity in site

- specific neoplasia;

**•** metastases;

**•** supporting informat ion 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 positive or false negative concerns;

**•** structure

- activity correlations; and

**•** in some cases, genetic toxicology

.

### 1

- Bromopropane , NTP TR 564 15

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

## NATIONAL TOXICOLOGY PROGRAM BOARD OF SCIENTIFIC COUNSELORS

TECHNICAL REPORTS REVIEW SUBCOMMITTEE

The members of the Technical Reports Review 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.

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 Immunology

University o f South Carolina School of Medicine

Columbia, SC 

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

Drug Safety Evaluation

Sanofi

- aventis

Alfortville, France



### Kenneth M. Portier, Ph.D.

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

### 16 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 

## SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMITTEE COMMENTS

NOTE: A summary of the Technical Reports Review Subcommittee’s remarks will appear in a future draft of this

report.

### 17

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# INTRODUCTION

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

- bromopropane produces

hydrog en 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 applications and in most commercial

products is stabilized to inhibit hydrolys is, and formulated to improve performance and enhance the useable life of

the product ( UNEP, 2001; HSDB

, 200 9).

### 18 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 

# P RODUCTION



# , U SE



# , AND 

# H UMAN 

# E XPOSURE

Industrial production of 1

- bromo propane generally involves reacting propanol with an excess of hydrogen bromide

gas. The principal product is 1

- bromopropane with small amounts of 2

- bromopropane (isopropyl bromide) and

other by

- products. Various modifications of the synthetic method are used to increase the purity of the

1- bromopropane, and distillation 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 pha rmaceutical intermediate or

process agent. U nited S tates production 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 intermediat e in the production of pesticides,

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 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 has a relatively high vapor pressure and is well

- suited

for use in cleaning equipment where it is repeatedly vaporized. Because it 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

applied adhesives containing 1

- bromopropane were used extensively by foam fabricating companies (Hanley et al

.,

2006) . Because 1

- bromopropane has a relatively short atmospheric half

- life (16 days)

, it is considered to have a

relatively low ozone depletion potential and has been marketed as a replacement for ozone depleting refrigerants

(e.g.

, chlorofluorohydrocarbons and hydroch lorofluorocarbons) ( Fed

. Regist

. , 2000)

.

Emissive use of 1

- bromopropane leads to substant ial 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 ccupational S afety and

H ealth (NIOSH) obtained personal breathing zone exposure data for workers at three foam fabrication plants that

used spray adhesive s containing 1

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

- weighted average ( TWA )

### 1- Bromopropane , NTP TR 564 19

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION exposures ranged from 18 to 381 ppm (mean = 142 ppm) (NIOSH

, 2002

, 2003) . In two of these plants the 8 -h our

TWA exposure concentrations 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 with [ 14 

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

- bromopropane 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 identified 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

- bromopropane, with m aximum

levels of S

- propyl glutathione detected 6 h ours after administrati on

. Exposure to 1

- bromopropane has been shown

to deplete 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- bromopropane 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 when 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)

.

The disposition and metabolism of 1

- bromopropane and the factors influencing them were extensively examined by

the N ational T oxicology Program (NT P) in male F344 /N rats and B6C3F1 mice following inhalation exposure

(800 ppm) or intravenous administration (5, 20, or 100 mg/kg) (Garner et al ., 2006). [ 13 

### C 3 

### ]-1- bromopropane and

[ 14 

### C] -1- bromopropane were co

- administered to enable characterization of urinary metabolites using nuclear

magnetic resonance spectroscopy and liquid chromatography coupled with either tandem mass spectrometry or

radiochromatography. By 4 hours f ollowing intravenous administration, rats and mice exhaled a majority of the

administered [ 14 

### C] -1- brom opropane 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%). The urinary metabolites characterized in rats and mice following both inhalation exposure and

### 20 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft intravenous administration were N

- acetyl

- S

- propylcysteine, N

- acetyl

- S

- (2

- hydroxypropyl)cys teine, N

- acetyl

3- (propylsulfinyl)alanine, 1

- bromo -2- hydroxypropane

- O

- glucuronide, N

- ace tyl

- S

- (2

- oxopropyl)cysteine, and

N

- acetyl -3- [(2

- oxopropyl)sulfinyl]alanine

.

In rats, but not in mice, the route of elimination and the metabolite distribution change d significantly as the dose

increased

, with t he percentage of dose excreted as volatile 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

dio xide decreased. An investigation of the molar ratio of 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 increased, the relative

proportion of N

- acetyl

- S

- propylcysteine and N

- acetyl

- S

- (2

- hydroxypropyl)cystei ne 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

c ytochrome 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

- bromopropane 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- hydroxypropane

,

N

- acetyl

- S

- (2

- hydroxypropyl)cysteine, was observed as the major urinary metabolite in wild type mice; the products

of direct conjugation of 1

- bromopropane with glutathione were reported ly insignificant. The ratio of glu tathione

conjugation to 2

- hydroxylation was increased five fold in the CYP 2E1 knockout mice relative to wild type mice. In

Sprague

- Dawley rats exposed to up to 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 dependence in the

metabolic mechanism in the rat liver was also proposed (Kim et al ., 1999)

. These results suggest that CYP2E1 is a

### 1- Bromopropane , NTP TR 564 21

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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 metabolism 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 wi th 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 investigated in rats by several routes of administration. The

inhalation LC

50 

### for 1

- bromopropane 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 r ats 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 ).

The LC

50 

### for 2

- bromopropane, a structurally related brominated alkane is repor ted as 31,171 ppm for a 4

- hour

inhalation exposure in mice ( ACGIH, 2005 ). The LD

50 

### for intraperitoneal injection of 2

- bromopropane 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 ).

### 22 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 23

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# R EPRODUCTIVE AND 

# D EVELOPMENTAL 

# T OXICITY



## Experimental An imals

1- Bromopropane 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 weight s and increased incidence s

of skeletal variations were observed in pups of dams exposed to 1

- bromopropane at 500 ppm or greater for

6 h ours /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 demonstrated that exposure to

400 or 800 ppm 1

- bromopropane for 12 weeks (8 hours/day, 7 days/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 significant 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 po tential for

1- bromopropane to produce adverse reproductive and developmental effects in humans (NTP, 2003). CERHR

concluded that there was convincing evidence for reproductive and developmental toxicity in experimental animals.

Evidence in humans was lim ited, but in the monograph, not e was made of a new case that was not available to the

expert panel indicating positive findings in women (altered menstruation) occupationally exposed to

1- bromopropane. 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 occupational exposure range (18 to

### 24 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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

- bromopropane

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

- bromopropane 8 h ours /d ay , 7 d ays per week for up to 7 weeks. Unexpectedly

,

1- bromopropane was found to be more neurotoxic than 2- bromopropane

. Electrophysiological changes (decrease d

maximum motor nerve conduction velocity and increased distal latency) and histopathologic 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 hindli mb 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

- brom opropane 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 degeneration in the peripheral nerve s, pre

- terminal axon swelling in the

gracile nucleus in the medulla oblongata, weight loss in the cerebrum , limb muscle weakness, distal latency

prolongation, and decrease of motor nerve conduction 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% b y weight) to strip metal for about 2 months. Nerve conduction studies indicated

a primary symmetric demyelinating polyneuropathy, and magnetic resonance imaging scans revealed evidence of

central nervous system (CNS) involvement. Subsequently, neurologic al signs of toxicity were reported in three

workers at a foam fabrication factory where they used a spray adhesive containing 1

- bromopropane (Ichihara et al

.,

### 1- Bromopropane , NTP TR 564 25

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 2004b) . The workers complained of numbness in the feet, thighs, and buttocks, and reduced sensations of vibration

in the feet suggesting peripheral neuropathy. Other symptoms such as headache, dizz iness, and memory loss

suggested adverse effects on the CNS. NIOSH reported that four additional workers in a different cushion

manufacturing company were hospitalized about a year earlier with similar neurological symptoms (NIOSH, 2002)

.

All four workers had elevated serum bromide le vels; however, exposure

- related effects on blood cell counts, male

reproductive functions, or nerve conduction velocity were not detected. NIOSH conducted 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 hand s 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, suggesting that 2

- bromopropane exposure caused polyneuropathy

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

.



# I MMUNOTOXICITY

1- Bromopropane was demonstrated to cause immunosuppression after oral administration to mice. A single

t reatment of female BALB/c mice with 1

- bromopropane by gavage significantly suppressed the antibody response

to T

- cell dependent antigen and the production of splenic intracellular IL

- 2 in response to conconavalin A ( Lee

et al ., 2007a ). Similarly, oral tre atment with 1,3

- dibromopropane significantly suppressed the antibody response to

T- dependent antigen in a dose

- dependent 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

- bromopropane was first indicated to be immunotoxic in a study designed to evaluate

reproductive toxicity in male rats. In this study, leukocyte numbers were significantly decreas ed in rats treated with

2- bromopropane concentrations that caused reproductive toxicity ( Yu et al ., 199 7 ). In a subsequent study ( Jeong

et al ., 2002) , 2

- bromopropane was shown to cause immunosuppression in rats following oral exposure to

1,000 mg/kg of 2

- bromopropane for 28 consecutive days. 2

- Bromopropane treatment significantly reduced numbers

of leukocytes, spleen cells, and thymic cells and suppre ssed the antibody response to SRBC s.

### 26 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft As part of the inhalation studies reported in this Technical Report, the potential immunotoxicity of 1

- bromopropane

was investigated because of the earlier reports of immunosuppression following oral exposure to 1

- bro mopropane

(Lee et al ., 2007a) and the immunotoxicity of structurally related chemicals. In an NTP study, 1-b romopropane was

found to cause some evidence of immunosuppression in F 344/N rats and B6C3F1 mice after inhalation exposure for

4 and 10 weeks ( Anderson et al ., 2009 ). 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

- bromopropane in experimental animals or epidemiology studies in

humans. However, NTP studies in rodents have demonstrated that several structurally related brominated

hydrocarbons are potent multisite carcinogens. Inhalation exposure of F344 /N rats to 1,2

- dibromoethane (ethylene

dibromide) caused carcinomas and adenocarcinomas of the nasal cavity and hemangiosarcomas of the circulatory

sy stem 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 adenomas and carcinomas in females

(NTP, 1982 a) . 1,2

- Dibromoethane was also carcinogenic in B6C3F1 mice

, causing alveolar/bronchiolar adenomas

and carcinomas in males and females; and hemangiosarcomas of the circulatory system, fibrosa r comas in

subcutaneous tissue, carcinomas of the nasal cavity, and adenocarcinomas of the mamm ary gland in females.

Inhalation exposure to 1,2

- dibromo -3- chloropropane caused carcinomas, squamous

- cell carcinomas, and

adenocarcinomas of the nasal cavity and squamous cell papillomas 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

papillary carcinomas in females and squamous cell carcinomas of the nasal cavity and alveolar/bronchiolar

adenomas and carcinomas in males and females. Dermal applicat ion of 2,3

- dibromo -1- propanol caused tumors at

multiple sites in F344/N rats and B6C3F1 mice (NTP , 1993 ).

### 1- Bromopropane , NTP TR 564 27

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# G ENETIC 

# T OXICITY

Published data on the 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 some mammalian test systems. Barber et al . (1981)

reported that 1

- bromopropane, tested over a concentration 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 concentrati on, was

unaffected by the addition of S9, thus indicating that 1

- bromopropane is a direct

- acting mutagen 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 letha l 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 significant treatment

- related increases in pre

or post

- implantation 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 induce

dominant lethal mutations in germ cells of male mice. Negative results were also reported from an earlier dominant

lethal study conducted in male Sprague

- Dawley rats treated with 400 mg/kg per day 1

- bromopropane 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 undosed females for 8 weeks

(Saito

- Suzuki et al ., 1982). Examination of uterine contents at gestation day 13 or 14 revealed sma ll 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 post

- meiotic

sperm (primarily early spermatids, mating weeks 5 and 6) (Saito

- Suzuki et al ., 1982).

### 28 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft NIOSH assessed DNA

- damage levels, measured by the alkaline single cell gel electrophoresis ( C omet) assay in

peripheral leukocytes of 64 workers from two facilities 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 (hi gh

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

significant 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 fac ility showed a small but not statistically

significant increase, the result of a significant (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 assessme nts of DNA damage in exposed workers, whole blood cultures from an unexposed

individual were treated in vitro with either 1

- bromopropane or 2

- bromopropane, and evaluated for DNA damage

using the Comet assay (Toraason et al ., 2006). Both 1

- bromopropane and 2- bromopropane induced significant

increases in DNA damage at the highest concentration 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 in duce

significant levels of DNA damage.



# R EGULATORY 

# S TANDARDS

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) regu lations and under the

Montreal Protocol on Substances that Deplete the Ozone Layer. In the late 1990s, 1

- bromopropane 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 adhesives

. As the United States Occupational Safety

and Health Administration ( OSHA ) began phasing in a more restrictive exposure standard for methylene chloride,

the use of 1

- bromopropane in adhesives increase d. In April 2000, OSHA ’ s final phase for instituting a workplace

exposure standard for methylene chloride took full effect

, leading companies to switch to 1

- bromopropane rather

### 1- Bromopropane , NTP TR 564 29

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION than install ventilation or other controls needed to reduce employee methylene chloride exposures to the requisite

25 ppm level. The increased production and use of 1

- bromopropane was a concern because it was not regulated to

protect workers, consumers, or the environment.

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 coat ings ( Fed

. Regist

. , 2007b)

. The American Conference of

Governmental Industrial Hygienists (2009) has recommended a threshold limit value 8

- hour time

- weighted average

for 1

- bromopropane of 10 ppm.



# S TUDY 

# R ATIONALE

1- Bromopropane was nominated by OSHA for evaluation of toxicokineti cs, genetic toxicity, reproductive and

developmental toxicity, neurotoxicity, and carcinogenicity. The nomination was based on the potential for

widespread occupational and environmental exposure to 1

- bromopropane and a lack of toxicity and carcinogenicit y

data. Because previous short

- term studies demonstrated 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 carcinogenicity of 1

- bromopropane

.

The 2

- week, 3

- month, and 2

- year studies were conducted in male and female F344/N rats and B6C3F1 mice to

evaluate the toxicity and carcinogenicity of 1

- bromopropane in association with inhalation exposure. Additional

s tudies were conducted to evaluate the absorption, distribution, metabolism, and excretion of 1

- bromopropane and

its potential immunotoxicity (Garner et al ., 2006; Anderson et al ., 2009)

.

### 30 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft

### 31

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# 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 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) (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 f or 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 t otal 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 determined to be

approximately 99%.

For lot 1581313004, Karl Fischer titration indicated a water content of 119 ppm. Elementa l analyses for carbon,

hydrogen, nitrogen, and sulfur were in agreement with the theoretical values for 1

- bromopropane; residual HBr was

### 32 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 bulk chemical was consistent throughout the sampled metal

drums, and the overall purity of lot 1581313004 was determined to 99. 9 % or greater

.

To ensure stability, the 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 , and no degradation of the bulk chemical was detected.



# V APOR 

# G ENERATION AND 

# E XPOSURE 

# S YSTEM

A preheater was necessary for the 2

- week and 3

- month studies. 1- Bromopropane was pumped t hrough a preheater

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 e ach 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 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 va por concentrations could be maintained throughout the chamber with the catch

pans in place. The total active mixing volume of each chamber was 1.7 m 3 

### . A small particle detector was used with

### 1- Bromopropane , NTP TR 564 33

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION and without animals in the exposure chambers to ensure that 1-b romopropane vapor, and not aerosol, was produced.

No particle counts above the minimum resolvable level (approximately 200 particles/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 exposure 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

were connected to the exposure chamber relative humidity sampling lines 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 allowed

digital measurement of sample flow. Summaries of the chamber vapor concentrat ions 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

concentration data to data from grab samples that were collected with activated coconut charcoal gas sampling

tubes, extracted with methylene chloride con taining 1

- bromobutane as an internal standard, and analyzed using an

off

- line gas chromatograph. Known volumes of chamber atmosphere were sampled at a constant flow rate ensured

by a calibrated critical orifice. The off

- line gas chromatograph was calibra ted 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 with out 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 beginning of vapor generation (T 90 

### ) and the time for the chamber concentrati on to

### 34 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 degradation impurities were

conducted periodically throughout the studies by GC. Chamber uniformity was maintained and no degradation was

detected.



# 2-W EEK 

# S TUDI ES

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 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 were 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 during exposure periods; water w as available ad libitum . Rats and mice

were housed individually . Clinical findin gs 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 summarized in Table 1.

Necropsies were performed on all animals . The heart, right kidney, liver, 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 to a thickness

of 4 to 6 μ m, and stained with hematoxylin and eosin. H istopathologic examinations were performed on all

chamber control and 2,000 ppm animals . The lung and nose of rats and mice; th e mediastinal lymph node, sciatic

### 1- Bromopropane , NTP TR 564 35

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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- bromopropane and to determine 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 selected for parasite

evaluation and gross observation for evidence 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 we re 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 anesthetized 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

chemistry analyses; blood was collected from the retroorbital sinus of mice at the end of the study for hematolo gy

### 36 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 volume; mean cell hemoglobin; and

mean cell hemoglobin concentra tion were determined using an Abbott Cell

- Dyn 3700 hematology 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 7100 Aerospray Slide Stainer (Wescor, Inc., Logan, UT). Leuko cyte

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 chemistry analyses were placed in tubes containing

separator gel and allowed to clot. After clot retraction occurred, the samples were centrifuged, and the serum was

aliquoted for assay of serum chemistry analytes usin g 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 collected for sperm motility and vaginal cytology evaluations on

rats and mice exposed to 0, 12 5 (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. Relative numbers of

leukocytes, nucleated epithelial cells, and large squamous epithelial cells were determined and used to ascertain

estrous cycle stage (i.e., diestrus, proestrus, estrus, and metestrus). Male animals were evaluated for sperm count

and motility. The left testis and left epididymis were isolated and weighe d. The tail of the epididymis (cauda

epididymis) was then removed from the epididymal body (corpus epididymis) and weighed. Test yolk (rats) or

modified Tyrode’s buffer (mice) was applied to slides and a small incision was made at the distal border of th e

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 esti mates, each left cauda epididymis was placed in buffered saline solution. Caudae were finely

minced, and the tissue was incubated in the saline solution and then heat fixed at 65° C. Sperm density was then

determined microscopically with the aid of a hemacytometer. To quantify spermatogenesis, the testicular spermatid

### 1- Bromopropane , NTP TR 564 37

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION head count was determined by removing the tunica albuginea and homogenizing the left testis in phosphate

- buffered

saline containing 10% dimethyl sulfoxide. Homogenization

- resistant spermat id 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 solution 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 eo sin. Complete histopathologic 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 histopathology 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 assessment

pathologists were resolved. 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 and 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

.

### 38 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 

## 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 parasite

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 hou sed individually . Feed was available 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 daily. Clinical findings 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 initially, weekly for

the first 13 weeks, then every 4 weeks through week 93, every 2 weeks thereafter, and a t 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

b uffered formalin (eyes were fixed in Davidson’s solution for up to 72 hours 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 f or microscopic examination. For all paired organs (e.g., adrenal gland, kidney, ovary),

samples from each organ were examined. Tissues examined microscopically 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 slides, paraffin blocks, and residual wet tissues were sent to the

### 1- Bromopropane , NTP TR 564 39

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION NTP Archives for inventory, slide/block match, and wet tissue audit. The slides, individual anima l 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 counts were verified, and the histotechnique 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 lung 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 inconsistencies in the diagnoses made by the laboratory and quality

assessment pathologists. Representative histopathology slides containing examples of lesions related to chemical

administration, examples of disagreements in diagnoses between the laboratory and quality assessment pathologists,

or lesions of general interest were presented by the coordinator to the PWG for review. The PWG consisted of the

qua lity assessment pathologist and other pathologists experienced in rodent toxicologic pathology. This group

examined the tissues without any knowledge of dose groups or previously rendered diagnoses. When the PWG

consensus 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

ta ken through the level of the incisive papilla 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 of Levels I and II (squamous epithelium) and

the dorsal meatus of Level II (olfactory epithelium). Level III is lined almost entirely by olfactory epithelium,

except for the ventral meatus, which is lined by resp iratory epithelium.

### 40 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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=maxilloturbinate; NT=nasoturbinate; ET=ethmoid turbinates

### 1- Bromopropane , NTP TR 564 41

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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

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 weeks 18 to 19 weeks Rats: 110 weeks

Mice: 109 to 111 weeks



### Size of Study Groups

5 males and 5 females Rats: 10 males and 10 females

Mice: 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

### 42 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 fluorescent 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 ra ts) 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 thereafter, and at the

end of the studies.

### 1- Bromopropane , NTP TR 564 43

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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; hemog lobin; erythrocyte, nucleated

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 histopa thology 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,

esoph agus, 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,

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. In addition, the liver,

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 addition 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 (c ecum, colon,

rectum), small intestine (duodenum, jejunum,

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

nodes (mandibular, mesenteric, bronchial,

mediastinal), mammary gland, nose, ovary,

pancreas, pancreatic islets, parathyroid gland,

pituitary gland, preput ial 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.

### 44 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 weighed.

Vaginal samp les 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 cycle

stages and estrous cycle length were

evaluated. None



# S TATISTICAL 

# M ETHODS



## Survival Analyses

The probability of survival was estimated by the product

- 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 analyses for possible dose

- related effects on survival used Cox’s

(1972) method for testing two groups for equality and Tarone’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 calculation of statistical significance, the incidences of most neoplasms (Tables A2,

B2, C2, and D2) and al l nonneoplastic lesions are given as the numbers of animals affected at each site examined

microscopically. However, when macroscopic examination was required to detect neoplasms in certain tissues (e.g.,

mesentery, pleura, peripheral nerve, skeletal musc le, tongue, tooth, and Zymbal’s gland ) before microscopic

### 1- Bromopropane , NTP TR 564 45

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION evaluation, the denominators consist of the number of animals that had a gross abnormality

. W hen neoplasms had

multiple potential sites of occurrence (e.g., leukemia or lymphoma), 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 method

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 animals 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

- response procedure that

modifies the Cochran

- Armitage linear trend test to take survival differences into account. More specifically, this

method modifies the denominator in the quantal estimate of lesion incidence to approximate more closely the total

number 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 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 function 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 evaluation of neoplasm on set 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 the 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 differential mortality, was accommodated by adjusting the variance of the Poly

- 3 statis tic as

recommended by Bieler and Williams (1993).

### 46 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft Tests of significance included pairwise comparisons of each exposed group with controls and a test for an overall

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 Variables

Two approaches were employed to assess the significance 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 analyz ed with the parametric multiple comparison procedures of Dunnett (1955) and

Williams (1971, 1972). Hematology, clinical chemistry, spermatid, and epididymal spermatozoal data, which have

typically skewed distributions, were analyzed using the nonparametri c multiple comparison methods of Shirley

(1977) (as modified by Williams, 1986) and Dunn (1964). Jonckheere’s test (Jonckheere, 1954) was used to assess

the significance of the dose

- related trends and to determine whether a trend

- sensitive test (Williams’ or Shirley’s

test) was more appropriate for pairwise comparisons 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 outlier 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 conformance 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 measurement s across exposure concentrations

. Proportions

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 matrices among exposure groups and between the control group and each

exposed group was tested using chi

- square statistics.

### 1- Bromopropane , NTP TR 564 47

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

## 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. However, historical control data are often helpful in interpreting

potential treatment

- related effects, particularly for uncommon or rare neoplasm types. For meanin gful comparisons,

the conditions for studies 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 incorporated a new diet

(NTP

- 2000) t hat 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 compl eted within the most recent 5

- year period. A second potential source of

variability is route of administration. 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 administration are

included for comparison, 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 Regulat ions (21 CFR, Part 58). In addition, as records from the 2

- year studies were submitted

to the NTP Archives, these studies were audited retrospectively by an independent quality assurance contractor.

Separate audits covered completeness and accuracy of th e pathology data, pathology specimens, 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 comments were resolved or

otherwise 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 mutations in

various strains of Salmo nella 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 numerical chromosomal alterations and are formed when acentric fragments or

### 48 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft whole chromosomes fail to incorporate into either of two daughter nuclei during cell division (Schmid, 1975;

Heddle et al. , 1983). The protocols for these studies and the results are given in Appendix E.

The genetic toxicity studies have evolved from an earlier effort by the NTP to develop a comprehensive database

permitting a critical anticipation of a chemical’s carcinogenicity in experimental animals based on numerous

considerations, including the mole cular structure of the chemical 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 mechanisms of chemical

- induced DNA da mage based on the relationship between electrophilicity

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

1985). However, it should be noted that not all cancers arise through genotoxic mechanism s.

DNA reactivity combined 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 sho wn to be the most predictive 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 th e predictivity of the Salmonella test alone. However, these other

tests can provide useful information 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 hav e high predictivity 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 positive results in rodent carcinogenicity studies (Witt et al. , 2000) . Because o f the

theoretical and observed associations between induced genetic damage and adverse effects in somatic and germ

cells, the determination of in vivo genetic effects is important to the overall understanding of the risks associated

with exposure to a part icular chemical. Most organic chemicals that are identified by the International Agency for

Research on Cancer as human carcinogens, other than hormones, are genotoxic. The vast majority of these are

### 1- Bromopropane , NTP TR 564 49

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION detected by both the Salmonella assay and rodent bone marrow cytogenetics tests (Shelby, 1988; Shelby and Zeiger,

1990).

### 50 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft

### 51

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# RE SULTS

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 h ind limb splaying was observed in some

2,000 ppm rats aft er 1 week of exposure to 1

- bromopropane

. This was a transient change and the animals showed

marked improvement after 1 day.

T ABLE 

### 2

Survival and Body Weights of Rats in the 2

- Week Inhalation Study of 1

- Bromopropane

Final Weight Concentration

a 

### Mean Body Weight b 

### (g) Survival Relative to Controls (ppm) 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 98

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

2 ,0 00 5/5 98 ± 4 12 3 ± 4** 25 ± 2 ** 73



### Female

0 5/5 87 ± 3 12 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 10 0

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

### 52 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft The absolute right kidney weights of 1,000 ppm males and all exposed groups of females we re 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. Relative liver weights were significantly increased in 500 ppm or greater males and all exposed

groups of females.

M icroscopic lesions related to 1

- bromopropane exposure occurred in the nose. I ncidences of minimal to mild

suppurativ e 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 1,000 and 2,000 ppm females (Table 3).

Sciatic nerve and spinal cord were examined microscopically, but no lesions were identified.

Exposure Concentration Selection Rationale : Based on decreased body weights and indications of neurotoxicity at

2,000 ppm in the 2

- week study, 1

- bromopropane exposure 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 No nneoplastic 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 53

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# 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.

T ABLE 

### 4

Survival and Body Weights o f Rats in the 3

- Month Inhalation Study of 1

- Bromopropane

Final Weight Concentration

a 

### Mean Body Weight b 

### (g) Survival Relative to Controls (ppm) Initial Final Change (%)

Male

0 10 /10 99 ± 2 345 ± 7 2 46 ± 6

62.5 10/10 102 ± 2 34 6 ± 5 2 44 ± 5 10 0

125 10/10 1 00 ± 2 34 8 ± 6 24 8 ± 5 101

250 10 /10 10 1 ± 3 350 ± 7 2 50 ± 6 10 1

500 10 /10 99 ± 2 344 ± 10 2 45 ± 10 100

1,000 10 /10 1 00 ± 2 305 ± 6** 205 ± 4 ** 88



### Female

0 10 /10 88 ± 1 202 ± 3 11 4 ± 3

62.5 10 /10 92 ± 3 20 3 ± 5 1 11 ± 3 101

125 10/10 88 ± 2 20 7 ± 4 1 20 ± 4 10 3

250 10/10 91 ± 2 20 9 ± 2 11 9 ± 3 10 4

500 10/10 86 ± 2 20 5 ± 3 119 ± 3 10 2

1,000 10 /10 88 ± 2 1 90 ± 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.

### 54 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft F IGURE 

### 3

G rowth Curves for Rats Exposed to 1

- Bromopropane

by Inhalation for 3 Months

### 1

- Bromopropane , NTP TR 564 55

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION There were no changes in hematology endpoints that were considered related 1

- bromopropane exposure (Table F1).

There were early, transient decreases in albumin and total protein concentrations and alanine aminotransferase

activit ies in most expos ed groups of male and female rats. These transient decreases may have be en associated with

effects of 1

- bromopropane on hepatic protein metabolism. Additionally, sorbitol d ehydrogenase (SDH) activit y w as

increase d at day 23 and at the end of the study in females exposed to 1,000 ppm and at the end of the study in males

exposed to 500 or 1,000 ppm. 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). T he absolute spleen weights of females exposed to 125 ppm or

greater and th e relative spleen weight of 1,000 ppm females were greater than those of the chamber controls. In

addition, the absolute and relative right kidney weights of 1,0 00 ppm females were greater than those of the chamber

controls.

There were significant exposure concentration

- related decreases in sperm motility in male rats exposed to 250 ppm

or greater (6.7%, 10.1%, and 27.7% in the 250, 500, and 1,000 ppm groups, respectively)

, and there were significant

decreases in the number of sperm per gram cauda a nd the total sperm per cauda (25.2% and 3 6.8%, respectively) , as

well as significant de creases in the absolute weights of the cauda (14%) and left epididymis (1 9 %) of 1,000 ppm

male rats (Table H1). Female rats in each of the exposure groups evaluated dif fere d significantly from the chamber

controls in the relative amount of time spent in the various estrous cycle stages , with each exposed group spending

significantly 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 hepatocellular

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 ep atocellular vacuolar degeneration was significantly increased in 1,000 ppm females.

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

### 56 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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



### population of enlarged, pale

- staining, degenerative, “ balloon

- like” cells admixed with low numbers of necrotic

hepatocytes. 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, supp urative, prostatic inflammation 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 F3 44/N rats, the biological significance 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 inhalation 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 compromise the 2

- year study.

### 1

- Bromopropane , NTP TR 564 57

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# 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 w as significantly less than that of the chamber control group

.

T he majority of the early deaths in the 500 ppm males w ere attributed to various types of neoplasia, none of which

w ere treatment related. However, in nine of these males, the cause of death w as attributed to inflammation in

various organs, all of which contained Splendore

- Hoeppli material. Survival of exposed groups of females

decreased with increasing exposure concentration

, but survival in each exposed group was not significantly different

from that of 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) 70 2 692 683 637

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

### 58 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft F IGURE 

### 4

Kaplan

- Meier Survival Curves for Rats

Exposed to 1- Bromopropane by Inhalation for 2 Years

### 1- Bromopropane , NTP TR 564 59

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

## 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 controls

throughout the study ( Tables 7 and 8 ; Figure 5 ). 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/v entral ulcer/ abscess (2/50, 7/50, 6/50, 20/50). No clinical findings related to exposure to 1

- bromopropane

were observed in females

. In male and female rats exposed to 1

- bromopropane, there was an exposure

- related

increased incidence of soft, pale

- yellow to green, variably sized nodules . These lesions were predominantly located

in the nose and/or skin, but other sites included bone, ear, Harderian gland, larynx, lung, muscle, peritoneum,

preputial gland, and prostate gland. The incidence s of these lesio ns 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 inflammation , many with Splendore

Hoeppli material.

### 60 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### 7

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

- Year Inhalation Study of 1- Bromopropane

Day s Chamber Control 1 25 ppm 25 0 ppm 5 00 ppm

on Av. Wt. No. of Av. Wt. Wt. (% of No. of Av. Wt. Wt. (% of No. of Av. Wt. Wt. (% of No. of

Study (g) Survivors (g) controls) Survivors (g) controls) Survivors (g) controls) 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 6 244 106

14

- 52 420 440 10 5 4 37 104 430 103

53

- 10 3 512 547 10 7 538 105 506 99

### 1- Bromopropane , NTP TR 564 61

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### 8

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

- Year Inhalation Study of 1- Bromopropane

Day s Chamber Control 1 25 ppm 25 0 ppm 5 00 ppm

on Av. Wt. No. of Av. Wt. Wt. (% of No. of Av. Wt. Wt. (% of No. of Av. Wt. Wt. (% of No. of

Study (g) Survivors (g) controls) Survivors (g) controls) Survivors (g) controls) 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

### 62 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft F IGURE 

### 5

Growth Curves for Rats Exposed to 1

- Bromopropane

by Inhalation for 2 Years

### 1- Bromopropane , NTP TR 564 63

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

## Pathology and Statistical Analyses

This section describes the statistically significant or biologically noteworthy changes in the incidences of malignant

mesothelioma and neoplasms or nonneoplastic lesions of the large intestine, skin, pancreatic islet s, nose, larynx,

t rachea, and lung

. Summaries of the incidences of neoplasms and nonneoplastic lesions

, statistical 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 mentio ned 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

significantly greater than that in the chamber control s ( 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 were not statistically

significant, the incidence in the 250 ppm group exceeded the historical control range s for inhalation studies and all

ro utes (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

thickene d by multiple layers of slightly enlarged neoplastic epithelial cells with basophilic cytoplasm and enlarged

nuclei. There were variable numbers of mixed inflammatory cells in the stroma around the glands and in the stalk.

No invasion of the submucosa la yer of the large intestine was observed.

### 64 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 number 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

c orresponding 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 neoplasm in animal group

i Value of statis tic cannot be computed.

j Historical incidence for inhalation studies : 0/ 350; all routes 3/ 1,350 ( 0.2 % ± 0.6 %), range 0%-2%

### 1- Bromopropane , NTP TR 564 65

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION Skin : In male rats, there were exposure concentration

- related increased incidences of keratoacanthoma;

keratoacanthoma 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 keratoacanthoma

and of keratoacanthoma or squamous cell carcinoma (combined ) in 250 and 500 ppm males were significantly

increased and exceeded the historical control range s for inhalation studies (Tables 10 and A3 b ). The incidences of

keratoacanthoma, basal cell adenoma, basal cell carcinoma, or squamous cell carcinoma (combine d) 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 squamous cell carcinoma in 500 ppm males

were not significantly i ncreased, but they exceeded the respective historical control ranges for inhalation studies

(Tables 10, A1

, and A3 b ).

In female rats, there were increased incidences of squamous cell papilloma, keratoacanthoma, basal cell adenoma, 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 respective historical control ranges for inhalation studies (Table s 10 and

B3b).

Keratoacanthomas were well

- demarcated, variably sized, crateriform masses in the dermis composed of squamous

epithelium that formed thick folds. The center of the mass was filled with abundant keratin. Squamous cell

carcinomas were characterized as masses composed of cords of pleomorphic squa mous cells infiltrating the dermis

and/or subcutis. There were varying amounts of keratin and fibrous connective tissue, which separated the cords of

squamous cells within the neoplasms. Basal cell adenomas were well

- demarcated masses composed of cords a nd

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 carcinomas were locally invasive and often

contained areas of necros is.

### 66 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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

Keratoacanthoma c

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

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

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

First i ncidence (days)

— h 729 (T) 488 562

Poly

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

Squamous Cell Carcinoma i 1 1 0 2

Keratoacanthoma or Squamous Cell Carcinoma j

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 k

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 l

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: 10/349 ( 2.9 % ± 3.7 %), range 0% -8 %; all routes: 66/1,39 8 ( 4.7 % ± 4.2 %), range 0%

- 16 %

d Number of animals with neoplasm per number 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 comparison s 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 neoplasm in animal group

i Historical incidence for inhalation studies: 1 /3 49 ( 0.3 % ± 0.8 %), range 0% -2 %; all routes: 8/1,398 ( 0.6 % ± 0 .9%), range 0% -2%

j Historical incidence for inhalation studies: 11/349 (3. 2 % ± 3.5%), range 0%

- 8%; all routes: 74 /1,398 ( 5.3 % ± 4. 1 %), range 0% -16%

k Historical incidence for inhalation studies: 1 9 /349 ( 5.5 % ± 4 .5%), range 0%

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

l Historical incidence for inhalation studies: 2 /3 50 ( 0.6 % ± 1. 0 %), range 0% -2 %; all routes: 16 /1,3 50 ( 1. 2 % ± 1.8 %), range 0% -6%

### 1- Bromopropane , NTP TR 564 67

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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 commo n site of

origin for mesotheliomas in F344/N rats ( Hall, 1990 )

, wa s the site of origin for all mesotheliomas in this study.

These mesotheliomas were characterized by numerous, complex, papillary structures composed 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 structures. There was often extensive invasion 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 Mesotheliom a 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 wit h 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

### 68 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft P ancreatic Islets : In male rats, the incidences of pancreatic 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 1 2 and A3 d ). The incidences of

pancreatic islet carcinoma were increased in the 125 and 250 ppm groups, but these increases were not statistically

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

increased 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.

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 ki ll

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. A negative trend is indicated by N

. f Not applicable; no neoplasm in animal group

g Historical incidence for inhalation studies: 17 /3 49 ( 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 /3 49 ( 10.6 % ± 4.8 %), range 6%-18 %; all routes: 119 /1, 394 (8.6% ± 4.0 %), range 0%- 18%

### 1- Bromopropane , NTP TR 564 69

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION Adenomas were usually discrete, well

- circumscribed masses of isle t 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 surrounding fibrous capsule or pancreatic

tissue.

Nose : I ncidence s of suppurative chronic inflammation in 500 ppm males and females were significantly greater

than those in the chamber controls (Table s 13 , A4, and B4 ). Suppurative chronic inflammation was characterized by

marked numbers of neutrophils with intralesional 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 -H oeppli material from suppurative inflammation without

Splendore

- Hoeppli material, which was observed in other organs. Occasionally

, neutrophils in the affected area

would invade the subjacent lamina propria. Application of special staining for bacteria i ndicated the presence of

Giemsa

- positive and Gram

- negative bacteria associated with 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 compared 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 lymphocyt es and macrophages, with fewer neutrophils within or

around the nasolacrimal duct, or within 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 e xposed groups of females compared to

that in the chamber controls (Table 13)

. Hyaline droplet accumulation was characterized by the presence of

intracytoplasmic, eosinophilic globules primarily within the olfactory epithelium and to a lesser extent the

### 70 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 71

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION respiratory epithelium. In general, this lesion was seen in many animals in the chamber control and exposed groups

and it occurred most often in Levels II and III.

T he incidences of glandular hyperplasia were significantly 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 submucosal glands

.

The incidences of respiratory epithelial hyperplasia were significantly increased in 125 and 500 ppm females

(Tables 1 3 and B4). Respiratory epithelial hyperplasia was located most often along the septum in L evel 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 significantl y increased in 500 ppm females

(Tables 1 3 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 cil iated

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

inflammation 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 inflamm ation in 250 ppm males and females and 500 ppm females were

significantly 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.

### 72 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 metaplasia was characterized by the replacement of normal

respiratory epithelium by squamous epithelial cells.

Trachea : In females, the incidences of chronic active inflammation and epithelial hyperplasia were significantly

increased in the 500 ppm group (Tables 13 and B4 ). Both lesions also occurred in males, but the incidences were

not significant ly 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.

Lung : 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 morphologically similar to that seen in other organs.

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), a nd 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.

### 1- Bromopropane , NTP TR 564 73

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# M ICE



# 2-W EEK 

# S TUD Y

All early deaths occurred during the first week of exposure; all 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 ). 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 decreased. A bnormal breathing, lethargy, and eye discharge were observed primarily during week 1

and occurred only in groups exposed to 500 ppm or greater.

T ABLE 

### 14

Survival and Body Weights of Mice in the 2

- Week Inhalation Study of 1

- Bromopropane

Final Weight Concentration

a 

### Mean Body Weight b 

### (g) Survival Relative to Controls (ppm) 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 10 2

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

### 74 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft In 1,000 ppm males, the absolute and relative heart weight s w ere significantly less than th ose of the chamber

controls while the absolute and relative liver weight s w ere significantly greater than th ose of the chamber controls

(Table G 3 ). A bsolute right kidn ey weights were significantly increased in 250 ppm or greater females, and the

relative right kidney weights were significantly 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 significantly decreased

.

M icroscopic lesions related to 1

- bromopropane exposure occurred in the lung, liver, and nose of males and fe males

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 vacuolization, 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 vacuolization were significantly increased in 1,000 ppm males and females. Sporadic incidences of

nasal les ions 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 Selection 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.

### 1- Bromopropane , NTP TR 564 75

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### 15

Incidences of Selected Nonneoplastic Lesions in Mice in the 2

- Week Inhalation Study of 1

- Bromoprop ane

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)

Cent r ilobular, 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

### 76 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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)

Cent r ilobular, 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 77

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# 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 observe d in moribund mice during week 1

.

There were no changes in hematology endpoints that were considered to be related to 1

- bromopropane exposure

(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 rela tive 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 Mean Body Weight b 

### (g) Survival a 

### Relative to Controls (ppm) Initial Final Change (%)

Male

0 10 /10 23.6 ± 0.3 39.6 ± 0. 8 1 6.0 ± 0.7

62.5 10/10 23.7 ± 0. 2 3 9.0 ± 0. 9 1 5.3 ± 0. 8 98

125 10/10 23.6 ± 0.1 3 9.6 ± 0. 6 1 5.9 ± 0. 6 10 0

250 9 /10 c 23.6 ± 0. 3 37. 9 ± 0. 7 1 4.4 ± 0.6 96

500 6 /10 d 23.4 ± 0.3 3 7.4 ± 1 .4 1 4.2 ± 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 3 3.0 ± 1.3 1 2.7 ± 1.1 10 7

125 10/10 20.4 ± 0.3 3 1.9 ± 0. 8 11.5 ± 0. 6 10 3

250 10/10 1 9.7 ± 0.3 30.2 ± 0.9 10.4 ± 0.9 98

500 5 /10 e 20.2 ± 0.1 3 1.3 ± 1. 4 11.0 ± 1. 5 10 1 

### Final Weight

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 eek s of death: 1, 1, 1, 2

e W eek of death s : 1

### 78 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft F IGURE 

### 6

Growth Curves for Mice Exposed to 1

- Bromopropane

by Inhalation for 3 Months

### 1- Bromopropane , NTP TR 564 79

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION There were slight increases in the cauda epididymis weights ( 9.1 % and 17.1 %) and significant decreases in sperm

motility (4.1% and 3.8%) 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

significantly 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 diestrus 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. I ncidences of cytoplasmic vacuolization of

the respiratory epithelium were significantly increased in 500 ppm males and females

, and the incidence of necrosis

of the respiratory epithelium was significantly increased in 500 ppm females (Table 1 7 ). O lfactory epithelium

necrosis and necrosis of the lateral nasal glands occurred in a few 500 ppm m ales

. A single incidence of

cytoplasmic vacuolization of the respiratory 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. Vacuolation consisted of

a single, large, clear vacuole displacing 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 airways 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 incidences of tracheal cytoplasmic vacuolization were significantly increased in 500 ppm males and

females (Table 17)

. Vacuolation of the laryngeal lining epithelium also occurred in one 62.5 ppm female that

survived to the end of the study. Vacuolation was characterized by cells with a large expansile clear space

occupying the majority of the cytoplasm. These cells ofte n had a pyknotic nucleus at the center, suggesting that

vacuolation progressed to necrosis. These cells had often lost their cilia. Hyperplasia 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 organization of the epithelium.

### 80 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 81

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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

### 82 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 epithelium 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 devoid of epithelial cells or lined by

attenuated epithelial cells. Cells remaining often contained colorless vacuoles displacing the cytoplasm.

Regeneration of bronchiolar e pithelium occurred in all groups of mice and was not treatment related

. Regeneration

was described as flatte ne d and disorganized epithelium with variation in nuclear size and location. Nuclei tended to

be apical rather than basilar. This lesion was grad ed as minimal in all cases.

In the liver, i ncidences of hepatocellular necrosis and hepatocellular degeneration were significantly increased in

500 ppm males and females; a single incidence of cytoplasmic vacuolization occurred in a 500 ppm male (Table 1 7 ).

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 massive in some animals ; it was characterized by uniform eosinophilic staining of

hepatocytes and loss of cell ular detail. Cytoplasmic degeneration and/or vacuolation, which w ere often seen

peripheral to the necrosis, consisted of swollen and foamy hepatocytes with one large or many tiny clear vacuoles

displacing the cytoplasm and with centrally located nuclei wi th condensed chromatin. Chronic inflammation

occurred in 500 ppm terminal 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, macrophages, large pigment

- laden macrophages, and occasional 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

.

Necrosis of the adrenal cortex (the zona fasciculata being most affected with extension into the zona glomerulosa to

a lesser degree) occurred in 500 ppm female mice that died early , and the incidence of this lesion was signific antly

increased in this group (Table 1 7)

. Chronic inflammation was noted at the corticomedullary junction in the adrenal

cortex of two 500 ppm females that survived to the end of the study.

### 1- Bromopropane , NTP TR 564 83

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION Exposure Concentration Selection Rationale : Based on mortality at the highest exposure concentration, changes in

organ weights, and the incidences of various nonneoplastic lesions in the 3

- month study, 1

- bromopropane exposure

concentrations selected for the 2

- year inhalation study in mice were 62.5, 125, and 250 ppm.

### 84 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 

# 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 control group s.

T ABLE 

### 18

Survival of Mice in the 2

- Year Inhalation Study of 1

- Bromopropane

62.5 ppm 125 ppm 25 0 ppm Control

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. 9 34 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 

### Chamber

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 85

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION F IGURE 

### 7

Kaplan

- Meier Survival Curves for Mice

Exposed to 1- Bromopropane by Inhalation for 2 Years

### 86 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 

## Body Weights and Clinical Findings

Mean body weights of exposed groups of male and female mice were similar to those of the chamber controls

throughout the study ( Figure 8 ; Tables 19 and 20 ). No clinical findings related to exposure to 1

- bromopropane were

observed.

### 1- Bromopropane , NTP TR 564 87

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION F IGURE 

### 8

Growth Curves for Mice Exposed to 1

- Bromopropane

by Inhalation for 2 Years

### 88 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### 19

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

- Year Inhalation Study of 1- Bromopropane

Day s Chamber Control 62.5 ppm 125 ppm 250 ppm

on Av. Wt. No. of Av. Wt. Wt. (% of No. of Av. Wt. Wt. (% of No. of Av. Wt. Wt. (% of No. of

Study (g) Survivors (g) controls) Survivors (g) controls) Survivors (g) controls) 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

6 91 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 89

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### 20

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

- Year Inhalation Study of 1- Bromopropane

Day s Chamber Control 62.5 ppm 125 ppm 250 ppm

on Av. Wt. No. of Av. Wt. Wt. (% of No. of Av. Wt. Wt. (% of No. of Av. Wt. Wt. (% of No. of

Study (g) Survivors (g) controls) Survivors (g) controls) Survivors (g) controls) 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

### 90 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 

## 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 analyses of primary neoplasms that occurred with an incidence of at least 5% in at

least one animal group, and historical incidences for the neoplas ms 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/bronchiolar ad enoma or carcinoma (combined) . T he incidences of

alveolar/bronchiolar adenoma or carcinoma (combined) 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 incidences 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 alveola r/bronchiolar carcinoma and alveolar/bronchiolar

adenoma or carcinoma (combined) in all exposed female groups exceed ed the respective historical control range s for

inhalation studies (Tables 2 1 and D3)

.

A lveolar/bronchiolar carcinomas varied from modera tely

- differentiated, circumscribed lesions to anaplastic, poorly

circumscribed, infiltrative lesions. They were variable in size and shape and their growth patterns were papillary or

solid with regions of squamous metaplasia, spindle cell differentiation, and necrosis. A lveolar/bronchiolar

adenomas were usually smaller, well

- differentiated, circumscribed lesions that often compressed the adjacent

parenchyma ; and m ost were papillary

.

Cytoplasmic vacuolization of bronchiolar epithelium occurred in all expo sed 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 th e 3- month inhalation study of 1

- bromopropane in

mice

.

### 1- Bromopropane , NTP TR 564 91

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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

### 92 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 mortality

g Observed incidence at terminal kill

h Beneath the cha m ber control incidence is the P value associated with the trend test. Beneath the exposed group incidence is the P value

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

- 3 test accounts for differential

mortality in animals that do n ot 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 %



### 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. Histologically, bronchiolar

regeneration was characterized by disorganized, elongated, and flattened epithelial cells lining terminal bronchioles

(Plates 8 through 10)

. Regenerative epithelial cells often had increased basophilia, lacked cilia, had varia ble

amounts of cytoplasm, and had increased nuclear to cytoplasmic ratios. Nuclei of regenerative cells tended to be

### 1- Bromopropane , NTP TR 564 93

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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 homogenous, hypereosinophilic cytoplasm.

Nose : There w ere exposure concentration

- related increase d incidence s o f cytoplasmic vacuolization of respiratory

epithelium in male and female mice ; the incidences in all exposed groups of males and in 125 and 2 5 0 ppm females

were significantly increased (Table s 21 , C 3 , and D4 )

. This lesion did not occur in chamber control mice.

Cytoplasmic vacuolization involved the respiratory epithelium on the lateral surfaces of nasoturbinates in L evels I

and II (Plate 1 1)

. Histologically, this lesion was characterized by large, solitary, clear vacuoles expanding the

cytoplasm of respiratory epithelial cells. Oftentimes, affected epithelial cells 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

.

Th ere w ere treatment

- related increase d incidence s of respiratory epithelial hyperplasia in the dorsal meatus(es) in

L evel I in mice of both sexes (Table s 21 , C 3 , and D4 ) ; the incidences in all exposed female groups and in 62.5 and

250 ppm males were signific antly increased

. Although present in chamber controls, the incidence and severity of

respiratory epithelial hyperplasia increased with increasing e xposure concentration, and male mice were more

severely affected than female mice. Histological criteria for respiratory epithelial hyperplasia included thickened

epithelium due to increased cellularity, submucosal epithelial invaginations with intraepithelial crypt

- like structures

(pseudoglands), 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 epithelia l hyperplasia was diagnosed

when the respiratory epithelium displayed focal or regional mucosal thickening with submucosal epithelial

invaginations in one or both dorsal meatus(es). Grade 2 (mild) respiratory epithelial hyperplasia was diagnosed

when at l east 2/3 of one or both dorsal meatus(es) displayed respiratory epithelium with submucosal epithelial

invaginations and/or epithelial papillary projections that together involved < 25% into the dorsal meatus(es)

.

Grade 3 (moderate) respiratory epithelial hyperplasia was diagnosed when two thirds to all of both dorsal

meatus(es) displayed respiratory epithelium with submucosal epithelial invaginations forming intraepithelial

### 94 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft crypt

- like structures (pseudoglands) and finger

- like papillary projections of epith elium that together involved 25 % to

50% into the dorsal meatus(es). Grade 4 (marked) respiratory epithelial hyperplasia was diagnosed when

approximately all of both dorsal meatuses had hyperplastic respiratory epithelium with tortuous, submucosal

invagina tions forming intraepithelial crypt

- like structures (pseudoglands) and finger

- like papillary projections of

epithelium 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 epithelium by ciliated, pseudostratified epithelium resembling respiratory

epit helium of the anterior nasal cavity; this lesion was often associated with submucosal inflammation and/or

glandular hyperplasia. The severity of the l esions w as graded minimal to mild based on the degree of involvement

of the dorsal meatuses and/or turbin ates.

The incidence of olfactory epithelial atrophy was significantly 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 signific ant (Tables 21 and C 3)

. This lesion consisted of segmental thinning of the olfactory epithelium

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 submucosal inflammation and/or glandular hyperplasia, and usually was

present within the dorsal meatuses

.

Larynx 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 cytoplasm of respiratory epithelial cells (Plate 1 2)

. A ffected epithelial cells often lacked

### 1- Bromopropane , NTP TR 564 95

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION cilia. Occasionally, nuclei of affected epithelial cells we re pyknotic. This lesion is similar to that described in the

3- month inhalation study of 1

- bromopropane in mice.



# G ENETIC 

# T OXICOLOGY

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

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 normochromatic erythrocytes were seen in peripheral blood of male or female B6C3F1 mice

exposed for 3 months to 62.5 to 500 ppm 1

- bromopropane via inhalation (Table E2). The percentage of

reticulocytes (polychromatic erythrocytes) in the peripheral blood of male and female mice was unaltered by

1- bromopropane exposure, suggesting a lack of chemical

- associated bone marrow toxicity.

### 96 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board DraftP 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 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&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&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 l ung of a

male B6C3F1 mouse exposed to 250 ppm 1

- bromopropane by inhalation for

2 years. Note the abundant necrotic 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

### 97

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# DISCUSSION AND CONCLUSIONS

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 suc h as vapor and immersion degreasing operations and critical cleaning of electronics and

metals. 1

- Bromopropane has also been proposed 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 carcinogenicity data. Because previous 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 carcinogenicity 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 the primary target site in these mice

, and severe centrilobular

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 hepat ic centrilobular

necrosis was present only 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 o ne 250 ppm male

. The centrilobular location of the lesions in the region of the liver that contains the

highest levels of P450 activity suggests that the hepatotoxicity may be due to a reactive metabolite of

1- bromopropane. In mic e, 1- bromopropane is oxi datively metabolized in the liver by cytochrome P450 2E1

(CYP2E1) to reactive metabolites that may be responsible for the toxicity of 1

- bromopropane (Garner et al ., 2007;

Liu et al ., 2009 ). 2- Oxo -1- bromopropane, 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 metabolites occurs primarily via

### 98 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft glutathione

- S

- transferase (GST)

- mediated conjugation with glutathione ( 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 ., 198 3). 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 glutathion e by inhibiting enzymes required

for glutathione synthesis. Alternatively, 1

- bromopropane metabolites may inhibit hepatocellular GST thereby

preventing glutathione conjugation. 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 occur red 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 50 0 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 dehydrogenase 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

- bromopropane than rats (Garner et al ., 2006). This

species difference in metabolic capacity may partially explain why extensive hepatotoxicity and early deaths

occurred in exposed mice and not in rats.

1- Bromopropane also caused lesions in the respiratory tract of mice, although these lesions were not considered as

severe as those in the liver. In the 2

- week study, respiratory tract lesions were most severe in mice that died early

after exposure to 500 ppm or greater . Necrosis and vacuolization of the nasal respiratory and olfactory epithelium

and prominent necr osis of the bronchiolar epithelium were present only in early death mice. In survivors exposed to

500 ppm or greater , lesions consisted primarily of regeneration of the olfactory epithelium and the bronchiolar

epithelium. In the 3

- month study, mild necro sis of the nasal respiratory and olfactory epithelium and minimal

### 1- Bromopropane , NTP TR 564 99

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION necrosis of the epithelium lining the larynx and trachea were observed primarily in 500 ppm mice that died early.

Minimal regeneration of the bronchiolar epithelium was observed in the majo rity 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 nasal epithelium cannot be r uled out.

In the 2

- week and 3

- month studies, r ats were less susceptible 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 inflamm ation were present in a few 1, 000 and 2

, 000 ppm male rats. There were no exposure

- related

nasal lesi ons in rats exposed for 3 months. Although the mechanism of this species difference is unclear, it may be

associated with differences in metabolic capabil ity in the nasal epithelium.

The potential immunotoxicity of 1

- bromopropane was investigated because of earlier reports of immunosuppression

following oral exposure to 1

- bromopropane (Lee et al ., 2007a), and the structurally related chemicals

2- bromopro pane ( Jeong et al

. , 2002 ), and 1,3

- dibromopropane (Lee et al ., 2007b). 1

- Bromopropane caused some

evidence of immunosuppression in F 344/N rats and B6C3F1 mice after inhalation exposure for 4 and 10 weeks

(Anderson et al ., 2009)

. Significant decreases in the IgM response to sheep red blood cells (SRBCs) were observed

in both species after exposure to 1

- bromopropane (Anderson et al ., 2009). In addition, s ignificant decreases in the

total number of spleen cells and T

- cells were obse rved 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 immunotoxicity of

1- bromopropane and 1,3

- dibromopropane may result from depletion of glutathione . Immunosuppres sion caused by

these two chemicals was correlated with chemical

- induced hepatotoxicity and decreases in liver and spleen

glutathione . Partial depletion of glutathione has been shown to affect immune function (effector phase of cytotoxic

T- cell responses a nd 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

- bromopropane may be due to r educed 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

### 100 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft mice. There was clear evidence that 1

- bromopropane exposure caused increase d in cidences of alveolar/bronchiolar

adenoma or carcinoma (combined) 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

alveolar/bronchiolar adenoma or carcinoma (combined) in female mice increased in a n exposure concentration

related manner

, were significantly greater than that in the cha mber controls at all exposure concentrations , and

exceeded the range in historical controls for inhalation 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 range in historical controls for inhalation studies and for all routes

. The incidences of alveolar/bronchiolar

carcinoma in 62.5 and 125 ppm female mice were significantly increased compared 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. Alveolar/bronchiolar

c arcinoma w as not observed in chamber control female mice . Chronic exposure to the structurally related

1,2

- dibromoethane also caused alveolar/bronchiolar adenomas and carcinomas 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 ra ts at various sites. There was

clear evidence that increased incidences of adenoma of the large intestine (colon and rectum ) were treatment related

in female rats. The increased incidence s of adenoma w ere exposure concentration related, and the incidence in the

500 ppm group was significantly greater than that in the chamber controls. There was some evidence that adenomas

of the large intestine in male rats were treatment related. Adenomas were found in the large intestine of two

250 ppm male rats (4%) and in one 500 ppm male rat (2%). Although 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. Ade nomas 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,

1984; Nigro, 1985). Oral treatment of rats with two brominated methanes ( bromodichloromethane and

### 1- Bromopropane , NTP TR 564 101

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION tribromomethane ) also resulted in significantly increased incidences of adenocarcinoma of the large intestine in both

male and female rats (NTP, 1987, 1989 ).

There was some evidence that chronic exposure to 1

- bromopropane caused an increase in the incidences of

neoplasms of the skin in male rats

, and skin neoplasms in fem ale rats may have been related to exposure

. In males,

t he combined incidences of all neoplas m s of epithelial origin increased in a significant exposure concentration

dependent manner. The incidences were significantly increased in all exposed groups of m ales 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 cell carcinoma. In

males, the incidences of keratoacanthoma increased with a positive trend ; the incidences in the 250 and 500 ppm

groups were significantly increased compared to chamber controls

, and exceeded the historical control range for

inhalation studies

. The increased incidence of keratoacanthoma is a concern because there is evidence that it can

progress to squamous cell carcinoma, a highly malignant tumor ( Elwell et al ., 1990 ). Squamous cell carcinoma is a

rare neoplasm occurring in only 0. 6 % of historical controls from a ll 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

un like 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

- bromopropane exposure was associated with an increased incidence of pancreatic islet adenoma

in male rats 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 studies (6%) , and t he incidences in exposed groups did not exceed the historical

control range s of inhalation studies or all routes.

### 102 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft The evidence for malignant mesothelioma in all organs was equivocal in male rats. The incidences of malignant

mesothelioma occurred with a positive 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 studies. The historical control range for malignant

mesothelioma in male rats for inhalation studies and by all routes is 0% to 6%. Although the incidence in the

500 ppm group was signific antly increased, it only exceeded the historical control range by one incidence.

An unusual nonneoplastic finding in rats exposed to 1

- bromopropane for 2 years was the presence of Splendore

Hoeppli reaction material associated with suppurative inflammati on. 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 mechanism of formation of Splendore

- Hoeppli material is unknown, but it is thought to

represent deposition of antigen

- antibody complexes and debris from host inflammatory cells (Hussein, 2008).

Alternatively, it may represent glycoproteins, lipid, and calcium derived from host leukocytes (Bhagavan et al

.,

1982). Histologically, Splendore

- Hoeppli material appears as strongly eosinophilic, amorphous material

surrounding or adjacent to the causative agent, typically fungi, helminthes or bacteria (Hussein, 2008). The

Splendore

- Hoeppli bodies appear microscopically as distinct r adiating, star

- like, or club

- shaped configurations. In

the United States, Splendore

- Hoeppli bodies are often seen in association with botryomycotic infections caused by

Proteus

, Escherichia coli or Pseudomonas aeruginosa ( Wenig et al ., 1996 ). In this study, cul 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 equally immunosup p ressed after

1- bromopropane exposure. Species differences in the presence 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 rat s was considered to be treatment related. The

incidence of lesions 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. Although the mechanism is not clear, immunosuppression in

1- bromopropane

- exposed rats may have contributed to the development of Splendore

- Hoeppli bodies. The

### 1- Bromopropane , NTP TR 564 103

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION development of botryomycosis and formation of Splendore

- Hoeppli bodies in humans may be facilitated by

immunosuppression as it has been associated with diabetes, immunoglobulin deficiency, cutaneous anergy,

corticosteroid treatment, and AIDS (Schlossberg et al

., 1980 ; Brunken et al ., 1983; Pa tterson et al ., 1987 ).

Because adequate data were available demonstrating the developmental and reproductive toxicity of

1- bromopropane, 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 studies, we conclude that

1- bromopropane 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 toxicity 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 conclusio n was that “there is serious concern for reproductive and developmental effects of 1

- bromopropane at

the upper end of the human occupational exposure range (18 to 381 ppm).” “Serious concern” is the highest level of

NTP conclusion regarding the possibilit ies that human development and reproduction might be adversely affected.



# 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 neoplasms of the skin. Increased incidences of malignant mesothelioma and pancreatic islet adenoma

may also have been related to 1

- bromopropane exposure. There was clear evidence of carcin ogenic activity of

1- bromopropane in female F344/N rats based on increased incidences of adenoma of the large intestine. Increased

incidences of neoplasms of the skin may also have been related to 1

- bromopropane exposure. There was no

evidence of carcino genic 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 carcinogenic activity of 1

- bromopropane in female

B6C3F1 mice based on increased incidences of alveolar/bro nchiolar neoplasms.

### 104 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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, the trachea of female rats and male and female mice, and the lung of mice.

Suppurativ e 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 14.

### 105

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

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### 112 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft

### A

- 1

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# APPENDIX A

SUMMARY OF LESIONS IN MALE R ATS

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

................................ ................................ ....... A

- 2

T ABLE 

### A2 Statistical Analysis of Primary Neoplasms in Male Rats

in the 2

- Year Inhalation Study of 1

- Bromopropane

................................ ................................ ...... A

- 6

T ABLE 

### A3 a Historical Incidence of Large Intestine (Colon or Rectum) Neoplasms

i n Control Male F344/N Rats

................................ ................................ ................................ .......... A

- 10

T ABLE 

### A3 b Historical Incidence of Skin Neoplasms i n Control Male F344/N Rats

................................ ....... A

- 11

T ABLE 

### A3 c Historical Incidence of Malignant Mesothelioma i n Control Male F344/N Rats

........................ A

- 12

T ABLE 

### A3 d Historical Incidence of Pancreatic Islet Neoplasms i n Control Male F344/N Rats

.................... A

- 1 2

T ABLE 

### A 4 Summary of the Incidence of Nonneoplastic Lesions in Male Rats

in the 2

- Year Inhalation Study of 1

- Bromopropane

................................ ................................ .... A

- 1 3

### A

- 2 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### A 1

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 A

- 3

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### A 1

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%)

### A

- 4 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### A 1

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 A

- 5

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### A 1

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

### A

- 6 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 A

- 7

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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

### A

- 8 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 A

- 9

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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.

### A

- 10 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 A

- 11

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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 Squamous Cell Carcinoma

Historical Incidence: Inhalation Studies

1

- Bromopropane (July, 2003) 0/50 0 /50 1 /50

Cumene (June, 2001) 4/50 1 /50 0 /50

Diethylamine (August, 2003) 0/50 0/50 0 /50

Methyl isobutyl ketone (May, 2000) 0/50 0/50 0 / 5 0

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 0 /50

Total (%) 10/349 (2.9%) 4 /349 ( 1.2 %) 1 /349 ( 0.3 %)

Mean 

### ± standard deviation 2.9% 

### ± 3.7% 1.2 .% 

### ± 1.1 % 0.3 % ± 0.8 %

Range 0%

- 8% 0 %

- 2 % 0 %

- 2 %



### Overall Historical Incidence: All Routes

Total (%) 66 /1,39 8 ( 4.7 %) 15 /1,39 8 ( 1.1 %) 8 /1,39 8 ( 0. 6 %)

Mean 

### ± standard deviation 4.7 % 

### ± 4.2 % 1.1 % 

### ± 1.4 % 0.6 % ± 0

. 9 %

Range 0%

- 1 6 % 0%

- 4 % 0%

- 2 %



### 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

Cumene (June, 2001) 4/50 5 /50

Diethylamine (August, 2003) 0/50 1 /50

Methyl isobutyl ketone (May, 2000) 0/50 0/50

α

- Methylstyrene (August, 2001) 1/50 2 /50

Propargyl alcohol (October, 2001) 4/49 5 /49

Tetralin (June, 2003) 1/50 5 /50

Total (%) 1 1 /349 ( 3.2 %) 1 9 /349 ( 5.4 %)

Mean 

### ± standard deviation 3.2 % 

### ± 3. 5 % 5.5 % 

### ± 4.5 %

Range 0%

- 8% 0%

- 10 %



### Overall Historical Incidence: All Routes

Total (%) 74 / 1,398 ( 5.3 %) 97 / 1,398 ( 6.9 %)

Mean 

### ± standard deviation 5.3 % 

### ± 4. 1 % 6.9 % 

### ± 4.9 %

Range 0%

- 1 6 % 0%

- 2 0 %

a Data as of April 29, 2009

### A

- 12 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 A

- 13

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### A 4

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

### A

- 14 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### A 4

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 A

- 15

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### A 4

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%)

### A

- 16 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### A 4

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 A

- 17

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### A 4

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%)

### A

- 18 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft

### B

- 1

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# APPENDIX B

SUMMARY OF LESIONS IN FEMALE RATS

IN THE 2

- YEAR INHALATION STUDY

OF 1

- BROMOPROPANE

T ABLE 

### B 1 Summary of the Incidence of Neoplasms in Fem ale Rats

in the 2

- Year Inhalation Study of 1

- Bromopropane

................................ ................................ ....... B

- 2

T ABLE 

### B 2 Statistical Analysis of Primary Neoplasms in Fem ale Rats

in the 2

- Year Inhalation Study of 1

- Bromopropane

................................ ................................ ....... B

- 6

T ABLE 

### B 3 a Historical Incidence of Large Intestine (Colon or Rectum) Neoplasms

i n Control Female F344/N Rats

................................ ................................ ................................ ....... B

- 10

T ABLE 

### B 3 b Historical Incidence of Skin Neoplasms i n Control Female F344/N Rats

................................ ... B

- 11

T ABLE 

### B 4 Summary of the Incidence of Nonneoplastic Lesions in Fem ale Rats

in the 2

- Year Inhalation Study of 1

- Bromopropane

................................ ................................ .... B

- 1 2

### B

- 2 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### B1

Summary of the Incidence of N e 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 B

- 3

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### B1

Summary of the Incidence of N e 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%)

### B

- 4 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### B1

Summary of the Incidence of N e 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 B

- 5

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### B1

Summary of the Incidence of N e 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

### B

- 6 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 B

- 7

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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

### B

- 8 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 B

- 9

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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 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 as sociated 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 r each 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

### B

- 10 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 B

- 11

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### B 3 b

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

### B

- 12 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 B

- 13

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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%)

### B

- 14 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 B

- 15

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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%)

### B

- 16 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft

### C

- 1

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# APPENDIX C

SUMMARY OF LESIONS I N MALE MICE

IN THE 2

- YEAR INHALATION STUDY

OF 1

- BROMOPROPANE

T ABLE 

### C 1 Summary of the Incidence of Neoplasms in Male Mice

in the 2

- Year Inhalation Study of 1

- Bromopropane

................................ ................................ ....... C

- 2

T ABLE 

### C 2 Statistical Analysis of Primary Neoplasms in Male Mice

in the 2

- Year Inhalation Study of 1

- Bromopropane

................................ ................................ ...... C

- 7

T ABLE 

### C 3 Summary of the Incidence of Nonneoplastic Lesions in Male Mice

in the 2

- Year Inhalation Study of 1

- Bromopropane

................................ ................................ .... C

- 10

### C

- 2 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### C1

Summary of the Incidence of N e 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 C

- 3

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### C1

Summary of the Incidence of N e 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)

### C

- 4 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### C1

Summary of the Incidence of N e 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

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Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### C1

Summary of the Incidence of N e 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%)

### C

- 6 1

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NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### C1

Summary of the Incidence of N e 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

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Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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

### C

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NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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

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Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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

### C

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NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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

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Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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%)

### C

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NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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

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Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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%)

### C

- 14 1

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NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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%)

### D

- 1

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# APPENDIX D

SUMMARY OF LESIONS I N FEMALE MICE

IN THE 2

- YEAR INHALATION STUDY

OF 1

- BROMOPROPANE

T ABLE 

### D 1 Summary of the Incidence of Neoplasms in Fem ale Mice

in the 2

- Year Inhalation Study of 1

- Bromopropane

................................ ................................ ....... D

- 2

T ABLE 

### D 2 Statistical Analysis of Primary Neoplasms in Fem ale Mice

in the 2

- Year Inhalation Study of 1

- Bromopropane

................................ ................................ ...... D

- 6

T ABLE 

### D 3 Historical Incidence of Alveolar/bronchiolar Neoplasms

in Control Female B6C3F1 Mice

................................ ................................ ................................ .... D

- 10

T ABLE 

### D4 Summary of the Incidence of Nonneoplastic Lesions in Fem ale Mice

in the 2

- Year Inhalation Study of 1

- Bromopropane

................................ ................................ .... D

- 1 1

### D

- 2 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### D1

Summary of the Incidence of N e 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 D

- 3

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### D1

Summary of the Incidence of N e 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%)

### D

- 4 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### D1

Summary of the Incidence of N e 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 D

- 5

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### D1

Summary of the Incidence of N e 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

### D

- 6 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 D

- 7

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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

### D

- 8 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 D

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Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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

cor responding 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

.

### D

- 10 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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

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Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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

### D

- 12 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 D

- 13

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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%)

### D

- 14 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 D

- 15

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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%)

### D

- 16 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft

### E-1

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# APPENDIX E

GENETIC TOXICOLOGY

B ACTERIAL 

### M UTAGENICITY 

### T EST 

### P ROTOCOL



### ....................................................................................................... E-2

M OUSE 

### P ERIPHERAL 

### B LOOD 

### M ICRONUCLEUS 

### T EST 

### P ROTOCOL



### ........................................................................... E-2

E VALUATION 

### P ROTOCOL



### ................................................................................................ ......................................... E-3

R ESULTS



### ................................................................................................ .................................................................... E-3

T ABLE 

### E1 Mutagenicity of 1- Bromopropane in Bacterial Tester Strains

....................................................... E-4

T ABLE 

### E2 Frequency of Micronuclei in Peripheral Blood Erythrocytes of Mice

Following Treatment with 1- Bromopropane by Inhalation for 3 Months

.................................... E-6

### E-2 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 

# 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 li ver) for

20 minutes at 37° C. Top agar supplemented with L



### - histidine and d

- biotin was added, and the contents of the 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 Laboratories 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. typhimurium 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 toxicity was observed. All trials were repeated

, and those that were conducted with S9

activation enzymes were repeated using the same or higher concentrations of S 9.

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 do se 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 positive 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 of 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 polychroma tic 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 st andard 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 variatio n. 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

fina l call of positive for micronucleus induction is preferably based on reproducibly 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 of statistical analyses, the

reproducibility of any effects observed, and the magnitudes of those effects.

### 1- Bromopropane , NTP TR 564 E-3

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# 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 chemical 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 a re not

simply combined into an overall result. Rather, all the data are critically 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 w ere seen

in peripheral blood of male or female B6C3F1 mice exposed for 3 months to 62.5 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

- bromopro pane exposure, suggesting a lack of chemical

- associated bone marrow toxicity.

### E-4 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### E1

Mutagenicity of 1- Bromopropane in Bacterial Tester Strains a



### n b 

### Reverta ts/Plate

Dose −S9 +hamster S9 +rat S9

Strain (μg/Plate) Trial 1 Trial 2 10% 30% 10% 30%

Study perfor med at BioRelianc e C orporation

TA100 0 150 ± 4 121 ± 16 104 ± 2 103 ± 9 124 ± 2 140 ± 8

33 115 ± 6 118 ± 6 121 ± 7

100 144 ± 10 1 24 ± 4 1 09 ± 12 104 ± 8 1 23 ± 2 133 ± 5

333 1 39 ± 7 1 27 ± 7 92 ± 8 108 ± 6 1 28 ± 10 117 ± 12

1,000 149 ± 7 1 28 ± 6 111 ± 7 98 ± 6 144 ± 5 1 03 ± 7

3, 333 c 117 ± 2 107 ± 11 c 98 ± 19 c 118 ± 6 c c c

1 0,000 Toxic Toxic 107 ± 7 123 ± 7

Toxic

Trial summary a Neg tive Negative Nega tive Nega tive Nega tive Nega tive

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 c 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

e 3, 333 6 ± 3 9 ± 1 c 8 ± 1 c 10 ± 1 c 8 ± 2 c 8 ± 0 c

10, 000 2 ± 1 c 6 ± 1 c 4 ± 1 c

Trial summary ve Negati Negative Negative Nega tive Nega tive Nega tive

Positive control 226 ± 10 199 ± 19 6 0 ± 6 13 7 ± 9 127 ± 13 80 ± 8



### TA97 0 134 ± 27 1 05 ± 12 156 ± 6 125 ± 9 1 61 ± 7 1 68 ± 13

33 1 30 ± 12 16 3 ± 13 1 56 ± 9

10 0 108 ± 9 1 21 ± 3 114 ± 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 ± 14 1 68 ± 5

1, 00 0 1 17 ± 4 125 ± 6 1 43 ± 7 166 ± 3 c 147 ± 8 180 ± 13

c 3, 333 86 ± 3 c 1 17 ± 7 1 13 ± 9 c 91 ± 16 c 1 22 ± 10 c 1 21 ± 13 c

1 0,000 40 ± 22 e 53 ± 3 c 70 ± 15 c

Trial summary Negative Nega tive Nega tive Nega tive Nega tive Nega tive

Positive control 354 ± 17 2 11 ± 4 988 ± 30 1,063 ± 1 38 1,084 ± 32 418 ± 16



### TA98 0 23 ± 3 14 ± 1 1 6 ± 1 29 ± 1 25 ± 2 31 ± 4

33 1 8 ± 2 21 ± 3 24 ± 3

10 0 23 ± 2 15 ± 2 24 ± 3 2 3 ± 2 22 ± 2 27 ± 3

333 1 7 ± 2 16 ± 1 14 ± 1 1 7 ± 2 21 ± 5 2 3 ± 2

1 ,0 00 1 9 ± 2 15 ± 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 19 ± 2 c 1 5 ± 1 c 15 ± 2 c

10, 000 Toxic c c 11 ± 2 13 ± 0

Trial summary Nega tive Nega tive Nega tive Nega tive Nega tive Nega tive

Positive control 340 ± 17 166 ± 3 875 ± 24 5 78 ± 17 302 ± 22 824 ± 28

### 1- Bromopropane , NTP TR 564 E-5

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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 Perfo rmed at SITEK Res earch Laborato ries (lot 1581313 004 used in the 2

- year s tudies)

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

10,0 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 16 ± 0 16 ± 1

,0 1 0 0

0 19 ± 2

1,50 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

c 5 ,00 0 2 ± 1 2 6 ± 4 2 6 ± 1

7 ,5 00 23 ± 3 2 0 ± 1

1 0,000 c c 6 ± 1 8 ± 2

Trial summary Negative Negative Negative Nega tive

Positive control 693 ± 26 693 ± 26 1, 163 ± 77 742 ± 37



### Escherichia co li WP2 uvrA /pK M 101

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 ± 10 124 ± 3 203 ± 14 20 1 ± 8

5 ,00 0 88 ± 3 c c 76 ± 17 192 ± 6 1 95 ± 10

7,5 00

1 0,000 1 71 ± 6 164 ± 25

149 ± 7 142 ± 4

Trial summary Negative Negative Nega tive Nega tive

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- phenyl enediamine (TA98), and methyl methanesulfonate ( E. coli ). The positive control for metabolic activation with all strains was

2- aminoanthracene.

e Slight toxicity and precipitate

### E-6 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft Table E2

Frequency of Micronuclei in Peripheral Blood Erythrocytes of Mice

Following Treatment with 1- Bromopropane by Inhalation for 3 Months a



### Exposure Number of Mice Micronucleated NCEs/ Compound Concentration with Erythrocytes val c b 

### P ue PCEs (%) b 

### 1,000 NCEs (ppm) Scored

Male

Air d 0 5 2.00 ± 0. 61 2. 52 ± 0. 30

1- Bromopropane 62.5 5 3.10 ± 0. 81 0. 06 15 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

500 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 8

d Chamber control

e Significance of micronucleated NCEs/1,000 NCEs tested by the one

- tailed trend test ; significant at P ≤0 .025

### F-1

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# 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

............................................................................................................................ F-2

T ABLE 

### F2 Hematology Data for Mice in the 3- Month Inhalation Study of 1- Bromopropane

...................... F-7

### F-2 1- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 F-3

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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*

### F

- 4 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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. 2 1 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 F

- 5

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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

### F

- 6 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 5 0 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 F-7

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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.

### F-8 1- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft

### G

- 1

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# APPENDIX G

ORGAN WEIGHTS

AND ORGAN

- WEIGHT

- TO

- BODY

- WEIGHT RATIOS

T ABLE 

### G 1 Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios for Rats

in the 2

- Week Inhalation Study of 1

- Bromopropane

................................ ................................ ..... G

- 2

T ABLE 

### G 2 Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios for Rats

in the 3

- Month Inhalation Study of 1

- Bromopropane

................................ ................................ .. G

- 3

T ABLE 

### G 3 Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios for Mice

in the 2

- Week Inhalation Study of 1

- Bromopropane

................................ ................................ .... G

- 4

T ABLE 

### G 4 Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratios for Mice

in the 3

- Month Inhalation Study of 1

- Bromopropane

................................ ................................ .. G

- 5

### G-2 1- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 G-3

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION T ABLE 

### G2

Organ Weights and Organ

- Weight

- to

- Body

- Weight Ratio s 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).

### G-4 1- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### G3

Organ Weights and Orga n- 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% m ortality.

### 1- Bromopropane, NTP TR 564 G-5

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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).

### G-6 1- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft

### H

- 1

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# APPENDIX H

REPRODUCTIVE TISSUE EVALUATIONS

AND ESTROUS CYCLE CH ARACTERIZATION

T ABLE 

### H 1 Summary of Reproductive Tissue Evaluations for Male Rats

i n the 3

- Month Inhalation Study of 1

- Bromopropane

................................ ................................ ... H

- 2

T ABLE 

### H 2 Estrous Cycle Characterization for Female Rats

in the 3

- Month Inhalation Study of 1

- Bromopropane

................................ ................................ .. H

- 2

T ABLE 

### H3 Summary of Reproductive Tissue Evaluations for Male Mice

i n the 3

- Month Inhal ation Study of 1

- Bromopropane

................................ ................................ .. H

- 3

T ABLE 

### H 4 Estrous Cycle Characterization for Female Mice

in the 3

- Month Inhalation Study of 1

- Bromopropane

................................ ................................ .. H

- 3

### H-2 1- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft T ABLE 

### H1

Summary of Reproductive Tissue Evaluations for Male Rats in the 3- Month Inhalation St udy

of 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 sig nificant 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 the 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 control 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 day s 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 exposed 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- Brom opropane, NTP TR 564 H-3

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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 con trol 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 groups and between

the chamber control group and each exposed group indicated that 5 00 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 control group.

### H

- 4 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft

### I

- 1

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# APPENDIX I

CHEMICAL CHARACTERIZ ATION AND

GENERATION OF CHAMBE R CONCENTRATIONS

P ROCUREMENT AND 

### C HARACTERIZATION OF 

### 1

- B ROMOPROPANE



### ................................ ................................ ......... I

- 2

V APOR 

### G ENERATION AND 

### E XPOSURE 

### S YSTEM



### ................................ ................................ ................................ ........ I

- 2

V APOR 

### C ONCENTRATION 

### M ONITORING



### ................................ ................................ ................................ ................... I

- 3

C HAMBER 

### A TMOSPHERE 

### C HARACTERIZATION



### ................................ ................................ ................................ ....... I

- 3

F IGURE 

### I 1 Infrared Absorption Spectrum of 1

- Bromopropane

................................ ................................ ........ I

- 5

F IGURE 

### I 2 1 

### H

- Nuclear Magnetic Resonance Spectrum o f 1

- Bromopropane

................................ ................... I

- 6

T ABLE 

### I 1 Gas Chromatography Systems Used in the Inhalation Studies of 1

- Bromopropane

.................... I

- 7

F IGURE 

### I 3 Schematic of the Vapor Generation and Delivery System

in the Inhalation Studies of 1

- Bromopropane

................................ ................................ ................... I

- 8

T ABLE 

### I 2 Summary of Chamber Concentrations in the 2

- Week Inhalation Studies

of 1

- Bromopropane

................................ ................................ ................................ ............................. I

- 9

T ABLE 

### I 3 Summary of Chamber Concentrations in the 3

- Month Inhalation Studies

of 1

- Bromopropane

................................ ................................ ................................ ............................ I

- 9

T ABLE 

### I 4 Summary of Chamber Concentrations in the 2

- Year Inhalation Studies

of 1

- Bromopropane

................................ ................................ ................................ .......................... I

- 10

### I

- 2 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 

# 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 in 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. (Maryland 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) of

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 g as chromatography (GC) by system A

(Table I1). In addition, Chemir/Polytech Laboratories, 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 analyse s 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 chromatogra ph (Dionex Corporation, Sunnyvale, CA) equipped with a conductivity detector and and

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 pr epared 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 p pm. 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 overall 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

colum n 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 det ermined 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 I

- 3

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION M etering valve s at th e manifold controlled flow to each chamber through individual Teflon ® 

### 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 throughou t the chamber with the catch

pans in place. The total active mixing 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

- bro mopropane vapor, and not aerosol, was produced. No particle counts 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) mi nutes during each 6

- hour exposure period using Hastelloy ®



### - C stream

- 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 lines 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 met er between the vacuum

regulator and the gas chromatograph allowed 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 supplie d by a standard generator (Kin

- Tek ; Precision Calibration Systems,

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 fro m grab samples that were

collected with activated coconut charcoal 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 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

- bromobutan e 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

the 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 terminat ed (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

ran ged from 9 to 10 minutes with animals present; T 10 

### values ranged 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 pre sent and from 9 to 11 minutes with animals present ; T 10 

### values 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 1 0 minutes was selected for the 3

- month and 2

- year studies.

The uniformity 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.

### I

- 4 1

- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 chamber. 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 persistence 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 mic e) and 1,000 ppm

(rats) chambers in the 3

- month studies, and the 250 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 minutes 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) minut es with animals present

.

Samples of the test atmosphere from the distribution 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 of

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 degrad ation products co

- eluted with

1

- bromopropane or the solvent, a second GC analysis 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

dete rmine 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 d uring 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 exposure 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 I-5

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION F IGURE 

### I1

Infrared Absorption Spectrum of 1

- Bromopropane

### I-6 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft F IGURE 

### I2

1 

### H- Nuclear Magnetic Resonance Spectrum of 1

- Bromopropane

### 1- Bromopropane, NTP TR 564 I-7

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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).

### I-8 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft F IGURE 

### I3

Schematic of the Vapor Generation and Delivery System in the Inhalation Studies

of 1

- Bromopropane

### 1- Bromopropane, NTP TR 564 I-9

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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

### I- 10 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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

### J-1

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# 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

............................................................................. J-2

T ABLE 

### J2 Vitamins and Minerals in NTP

- 2000 Rat and Mouse Ration

.......................................................... J-2

T ABLE 

### J3 Nutrient Composition of NTP

- 2000 Rat and Mouse Ration

............................................................ J-3

T ABLE 

### J4 Contaminant Levels in NTP

- 2000 Rat and Mouse Ration

.............................................................. J-4

### J-2 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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 J-3

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 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)

### J-4 1- Bromopropane , NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 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.

### K-1

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 

# APPENDIX K

SENTINEL ANIMAL PROG RAM

Methods

................................................................................................ ................................................................... K-2

Results

................................................................................................ ...................................................................... K-4

### K

- 2 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft 

# 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 heal th that occurs during the toxicologic evaluation of chemical compounds. Under

this program, the disease state of the rodents is monitored via serology on sera from extra (sentinel) animals in the

study rooms. These animals and the study animals are subje ct 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 studie s, 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. Bloo d 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 serology methods and

viral agents for which testing was performed are tabulated below; the times at whic h blood was collected during the

studies are also listed.

Method and Test Time of Collection



# R ATS

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 (rat coronavirus) Study termination

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

(rat coronavirus/sialodacryoadenitis virus) 1 week, study termination

Sendai 1 week, study termination

Immunofluorescence Assay

Parvovirus Study termination

### 1

- Bromopropane , NTP TR 564 K

- 3

Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION Method and Test Time of Collection



# R ATS (continued)



### 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

Parvovirus 6, 12, and 18 months, s tudy 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

### K

- 4 1

- Bromopropane, NTP TR 564

NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft Method and Test Time of Collection



# M ICE (continued)



### 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

MCMV Study termination

Parvovirus 6, 12, and 18 months, study termination

Polymerase Chain Reaction

Helicobacter species 18 months



# R ESULTS

All test results were negative.