summary of drinking water carcinogenicity study of 2 ......(study no.0613) summary of drinking water...
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Summary of Drinking Water Carcinogenicity Study
of 2-Methyl-1-Propanol
in B6D2F1 Mice
September 2009
Japan Bioassay Research Center
Japan Industrial Safety and Health Association
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PREFACE
The tests were contracted and supported by the Ministry of Health, Labour and Welfare of
Japan. The tests were conducted by Japan Bioassay Research Center (JBRC) and the report was
prepared by JBRC and peer reviewed by outside expert pathologist. Complete report was
submitted to Ministry of Health, Labour and Welfare of Japan on September 30, 2009.
This English Summary was translated by JBRC from Japanese complete report.
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(Study No.0613)
Summary of Drinking Water Carcinogenicity Study
of 2-Methyl-1-Propanol
in B6D2F1 Mice
Purpose, materials and methods
2-Methyl-1-propanol (CAS No. 78-83-1) is a colorless clear liquid with a melting point of
-108C and a boiling point of 107.9C. It is soluble in water (8.7 wt%), alcohol, and ether.
The carcinogenicity and chronic toxicity of 2-methyl-1-propanol were examined in
B6D2F1/Crlj mice. Groups of test animals were administered 2-methyl-1-propanol in their
drinking water for 2 years (104 weeks). Each group consisted of either 50 male or 50 female
mice. The drinking water concentrations of 2-methyl-1-propanol were 0, 5000, 10000 or 20000
ppm (w/w) for males, and 0, 2500, 5000 or 10000 ppm for females. The highest dose level was
chosen so as not to exceed the maximum tolerated dose (MTD), based on both growth rate and
toxicity in a previous 13-week toxicity study. 2-Methyl-1-propanol was analyzed for purity and
stability by both infrared spectrometry and gas chromatography before and after its use. The
concentrations of 2-methyl-1-propanol in the drinking water were determined by gas
chromatography at the time of preparation and on the 4th day after preparation, while stored at
room temperature. The animals were observed daily for clinical signs and mortality. Body
weight, water consumption and food consumption were measured once a week for the first 14
weeks and every 4 weeks thereafter. Animals found dead, in a moribund state, or surviving to
the end of the 2-year administration period underwent complete necropsy. Urinalysis was
performed near the end of the administration period. For hematology and blood biochemistry at
the terminal necropsy, surviving animals were fasted overnight and bled under deep ether
anesthesia. Organs and tissues were removed, weighed and examined for macroscopic lesions at
necropsy. The organs and tissues were then fixed and embedded in paraffin. Five μm thick
tissue sections were prepared and stained with hematoxylin and eosin and examined
microscopically. Incidences of neoplastic lesions were statistically analyzed by Fisher’s exact
test. Any positive dose-response trends of 2-methyl-1-propanol induction of neoplastic lesions
were analyzed by Peto’s test. Incidences of non-neoplastic lesions and urinalysis were analyzed
by the Chi-square test. Changes in body weight, food consumption, hematological and blood
biochemical parameters, and organ weights were analyzed by Dunnett’s test. The present studies
were conducted in accordance with the Organisation for Economic Co-operation and
Development (OECD) Good Laboratory Practice and with reference to the OECD Guideline for
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(Study No.0613)
Testing of Chemicals 451 “Carcinogenicity Studies”.
Results
No significant differences in survival rates were found between any of the groups
administered 2-methyl-1-propanol and their respective controls. The growth rates of the females
administered 10000 ppm were slightly suppressed toward the end of the administration periods,
and body weights at 104 weeks were 88% of the control. Food consumption in the 5000 ppm-
and 10000 ppm-administered females showed temporary decreases. Food consumption in the
5000 ppm-administered males also showed temporary decreases. Food consumption in the
10000-administered males decreased substantially, and consumption in the 20000-administered
males was decreased for most of the 2-year administration period. Water consumption was
decreased through the most of the 2-year administration period in males administered 2500 ppm
or above and in females administered 5000 ppm or above. No other clinical signs were observed.
No significant increases in the incidence of neoplastic or tumor-related lesions was found in
any of the 2-methyl-1-propanol-administered groups of either sex compared with their
respective controls; nor was there a change in any other parameter in any of the groups.
The no-observed-adverse-effect-level (NOAEL) of 2-methyl-1-propanol in the drinking water
was 20000 ppm (1796 mg/kg body weight per day) in males and 5000 ppm (664 mg/kg body
weight per day) for the endpoint of body weight in females.
Conclusions
There was no evidence of carcinogenic activity of 2-methyl-1-propanol in male or female
mice.
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(Study No. 0613)
Incidences of selected neoplastic lesions of male mice in the 2-year drinking water
carcinogenicity study of 2-Methyl-1-propanol
Dose (ppm) 0 5000 10000 20000 Petotest
Cochran-Armitage test
Number of examined animals
50 50 50 50
benign tumor lung bronchiolar-alveolar
adenoma 7 4 4 3
liver hepatocellular adenoma 12 7 14 7 malignant tumor lung bronchiolar-alveolar
carcinoma 4 10 7 5
lymph node
malignant lymphoma 13 6 6 6
liver hepatocellular carcinoma 6 9 6 6 histiocytic sarcoma 3 3 1 4 hemangiosarcoma 1 2 3 1
Incidences of selected neoplastic lesions of female mice in the 2-year drinking water
carcinogenicity study of 2-Methyl-1-propanol
Dose (ppm) 0 2500 5000 10000 Petotest
Cochran-Armitage test
Number of examined animals
50 50 50 50
benign tumor liver hepatocellular adenoma 3 1 4 1 hemangioma 3 3 3 1 pituitary adenoma 12 9 5 6 ovary cystadenoma 0 0 3 1 hemangioma 0 1 4 0 Harderian gland
adenoma 3 2 0 4
malignant tumor lung bronchiolar-alveolar
carcinoma 2 2 1 3
lymph node malignant lymphoma 12 18 19 16 uterus histiocytic sarcoma 8 14 7 14 Significant difference * : p≦0.05 ** : p≦0.01 (Fisher test) ↑: p≦0.05 increase ↑↑: p≦0.01 increase (Peto, Cochran-Armitage test) ↓: p≦0.05 decrease ↓↓: p≦0.01 decrease (Cochran-Armitage test)
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(Study No. 0613)
SELECTED TABLES TABLE C 1 BODY WEIGHT CHANGES AND SURVIVAL ANIMAL NUMBERS:
MALE TABLE C 2 BODY WEIGHT CHANGES AND SURVIVAL ANIMAL NUMBERS:
FEMALE TABLE C 3 BODY WEIGHT CHANGES: MALE TABLE C 4 BODY WEIGHT CHANGES: FEMALE TABLE D 1 FOOD CONSUMPTION CHANGES AND SURVIVAL ANIMAL
NUMBERS: MALE TABLE D 2 FOOD CONSUMPTION CHANGES AND SURVIVAL ANIMAL
NUMBERS: FEMALE TABLE D 3 FOOD CONSUMPTION CHANGES: MALE TABLE D 4 FOOD CONSUMPTION CHANGES: FEMALE TABLE E 1 WATER CONSUMPTION CHANGES AND SURVIVAL ANIMAL
NUMBERS: MALE TABLE E 2 WATER CONSUMPTION CHANGES AND SURVIVAL ANIMAL
NUMBERS: FEMALE TABLE E 3 WATER CONSUMPTION CHANGES: MALE TABLE E 4 WATER CONSUMPTION CHANGES: FEMALE TABLE F 1 CHEMICAL INTAKE CHANGES: MALE TABLE F 2 CHEMICAL INTAKE CHANGES: FEMALE TABLE G 1 HEMATOLOGY: MALE TABLE G 2 HEMATOLOGY: FEMALE TABLE H 1 BIOCHEMISTRY: MALE TABLE H 2 BIOCHEMISTRY: FEMALE
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(Study No. 0613)
TABLE I 1 URINALYSIS: MALE TABLE I 2 URINALYSIS: FEMALE TABLE K 1 ORGAN WEIGHT, ABSOLUTE: MALE TABLE K 2 ORGAN WEIGHT, ABSOLUTE: FEMALE TABLE L 1 ORGAN WEIGHT, RELATIVE: MALE TABLE L 2 ORGAN WEIGHT, RELATIVE: FEMALE TABLE M 1 HISTOPATHOLOGICAL FINDINGS: NON-NEOPLASTIC LESIONS: MALE: ALL ANIMALS TABLE M 4 HISTOPATHOLOGICAL FINDINGS: NON-NEOPLASTIC LESIONS: FEMALE: ALL ANIMALS TABLE P 1 NEOPLASTIC LESIONS-INCIDENCE AND STATISTICAL ANALYSIS: MALE TABLE P 2 NEOPLASTIC LESIONS-INCIDENCE AND STATISTICAL ANALYSIS: FEMALE TABLE R 1 CAUSE OF DEATH: MALE TABLE R 2 CAUSE OF DEATH: FEMALE
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(Study No. 0613)
FIGURES FIGURE 1 SURVIVAL ANIMAL RATE OF MALE MICE IN THE 2-YEAR
DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL FIGURE 2 SURVIVAL ANIMAL RATE OF FEMALE MICE IN THE 2-YEAR
DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL FIGURE 3 BODY WEIGHT CHANGES OF MALE MICE IN THE 2-YEAR
DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL FIGURE 4 BODY WEIGHT CHANGES OF FEMALE MICE IN THE 2-YEAR
DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL FIGURE 5 FOOD CONSUMPTION CHANGES OF MALE MICE IN THE 2-YEAR
DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL FIGURE 6 FOOD CONSUMPTION CHANGES OF FEMALE MICE IN THE
2-YEAR DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL FIGURE 7 WATER CONSUMPTION CHANGES OF MALE MICE IN THE 2-YEAR
DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL FIGURE 8 WATER CONSUMPTION CHANGES OF FEMALE MICE IN THE
2-YEAR DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL
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(Study No. 0613)
FIGURE 1 SURVIVAL ANIMAL RATE OF MALE MICE IN THE 2-YEAR DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL
FIGURE 2 SURVIVAL ANIMAL RATE OF FEMALE MICE IN THE 2-YEAR DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL
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(Study No. 0613)
FIGURE 3 BODY WEIGHT CHANGES OF MALE MICE IN THE 2-YEAR DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL
FIGURE 4 BODY WEIGHT CHANGES OF FEMALE MICE IN THE 2-YEAR DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL
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(Study No. 0613)
FIGURE 5 FOOD CONSUMPTION CHANGES OF MALE MICE IN THE 2-YEAR DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL
FIGURE 6 FOOD CONSUMPTION CHANGES OF FEMALE MICE IN THE 2-YEAR DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL
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(Study No. 0613)
FIGURE 7 WATER CONSUMPTION CHANGES OF MALE MICE IN THE 2-YEAR DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL
FIGURE 8 WATER CONSUMPTION CHANGES OF FEMALE MICE IN THE 2-YEAR DRINKING WATER STUDY OF 2-METHYL-1-PROPANOL
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