1-chlorobutane

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March - June 1979

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: comparable to guideline study with acceptable restrictions

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference 1

Reference Type:

publication

Title:

Unnamed

Year:

1986

Reference 2

Reference Type:

secondary source

Title:

1-chlorobutane, CAS No. 109-69-3

Author:

OECD SIDS

Year:

1997

Bibliographic source:

SIDS Initial Assessment Report for SIAM 6

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

yes

Remarks:

only necropsy and histologic examination were performed, other observations (e.g. food consumption, haematology, clinical biology) are not regarded; administration five days per week

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

1-chlorobutane

EC Number:

203-696-6

EC Name:

1-chlorobutane

Cas Number:

109-69-3

Molecular formula:

C4H9Cl

IUPAC Name:

1-chlorobutane

Details on test material:

- Name of test material (as cited in study report): n-butyl chloride
- Storage condition of test material: in the dark at 0 °C

Test animals

Species:

rat

Strain:

other: F344/N

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Portage, USA
- Age at study initiation: 7 weeks
- Weight at study initiation: males: 23 g; females: 18-19 g
- Housing: five per cage in polycarbonate cages (Lab Products, Rochelle Park, NJ, USA); Bedding: Aspen bed hardwood chips (American Exelsior Co., Baltimore,MD, USA) or Chips hardwood chips (Agway Inc., Syracuse, NY, USA)
- Diet: ad libitum Wayne Lab Blox pellets (Allied Mill, Chicago, IL; USA)
- Water: ad libitum (Automatic watering system; Edstrom Industries, Waterford, WI, USA)
- Acclimation period: 18 days


ENVIRONMENTAL CONDITIONS
- Temperature: mean 21.8 °C
- Humidity: 5% - 74% (average 40%)
- Air changes (per hr): 10
- Photoperiod: 12 hrs dark / 12 hrs light ( fluorescent light)


IN-LIFE DATES: March - July 1979

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Preparation were hand agitated for 10 sec and sealed in serum vials. Dose mixtures were stored at 4 °C for a maximum of 10 days.

VEHICLE
- Amount of vehicle (if gavage): 5 mL/kg bw/d

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Periodic analyses for n-butyl chloride in corn oil were performed by the testing and analytical chemistry laboratories. Duplicate 1 mL samples were extracted with methanol containing 2 mg/mL of n-amyl alcohol as an internal standard. Samples were analyzed by GC-FID. Individually spiked portions of undosed corn oil (5 or 6 concentrations bracketing the specified concentration range of the referee sample) were used to obtain standard data. The samples were determined from the linear regression equation computed from the standard data (peak area analysis).
Analyzed mixtures were within +/- 10% of the target concentration.

Duration of treatment / exposure:

13 weeks (90d)

Frequency of treatment:

5 d/w

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 30, 60, 120, 250 and 500 mg/kg bw/d
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Doses selected for the 13-week studies were based on weight gain depression and clinical signs observed in the 14-days studies.

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: two times per day

CLINICAL SIGNS
- Time schedule: recorded once per week

BODY WEIGHT: Yes
- Time schedule for examinations: once per week

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: No

CLINICAL CHEMISTRY: No

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

The following tissues were examined: gross lesions and tissue masses, mandibular lymph node, mammary gland, skin, salivary gland, sternebrae, thyroid gland, small intestine, colon, liver, prostate/testes or ovaries/uterus, lungs and bronchi, heart, esophagus, stomach, brain, thymus, trachea, pancreas, spleen, kidneys, adrenal glands, urinary bladder, pituitary gland, spinal cord (if neurologic signs present) and eyes (if grossly abnormal).

Tissues were preserved in 10% neutral buffered formalin, embedded in paraffin, sectioned and stained with hematoxylin and eosin.
After completion of the pathology examination, the slides, individual animal data records and summary tables were sent to an independent quality assurance laboratory for evaluation.

Statistics:

Data recording:
Data were recorded in the Cacinogenesis Bioassay Data System (Linhart et al., 1974). The data elements include the recommendations by the International Union Against Cancer (Berenblum, 1969)

Survival analyses:
The probability of survival was estimated by the product-limit procedure of Kaplan and Meier (1985) and is presented in the form of graphs. Statistical analyses for a possible dose-related effect on survival used the method of Cox (1972) for testing two groups for equality and Tarone's (1975) life table test for a dose-related trend. All reported P values for the survival analysis are two-sided.

Results and discussion

Results of examinations

Details on results:

CLINICAL SIGNS AND MORTALITY
6/10 males that received 500 mg/kg bw/d n-butyl chloride died before the end of the study, three of these because of gavage accidents.
Hyperactivity and convulsion on one or more occasions in male and female animals of the 250 and 500 mg/kg bw/d groups (250 mg/kg bw/d: 5/10 males and 2/10 females; 500 mg/kg bw/d: 9/10 males and 8/10 females). No animals of the lower dose group were hyperactive or convulsed.

BODY WEIGHT AND WEIGHT GAIN
The final mean body weights of males of the 250 and 500 mg/kg bw/d dose group were 11% or 20% lower than those of the vehicle control group. Females exposed to 250 and 500 mg/kg bw/d were 6% or 10% lower than that of the controls.

GROSS PATHOLOGY
Mild to moderate compound-related extramedullary hematopoiesis in the spleen in 3/10 males were observed that received 500 mg/kg bw/d. In two rats the severity was mild and in a third animal moderate. This lesion was not examined in the verhicle control group.

HISTOPATHOLOGY:
no histopathologic changes

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

120 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

2

Dose descriptor:

LOAEL

Effect level:

250 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: clinical signs; mortality; body weight;

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Body weights of rats in the thirteen-week gavage studies of n-butyl chloride

Dose (mg/kg bw/d)	Mean body weights (g)			Final weight relative to vehicle controls (%)
	initial^a	final	change^b
Males
0	131 +/- 2	299 +/- 4	+168 +/- 3	---
30	131 +/- 2	300 +/- 5	+169 +/- 5	100
60	130 +/- 2	290 +/- 4	+160 +/- 3	97
120	131 +/- 2	285 +/- 3	+154 +/- 4	95
250	131 +/- 2	265 +/- 4	+134 +/- 3	89
500	131 +/- 2	240 +/- 10	+113 +/- 9	80
Females
0	103 +/- 1	181 +/- 8	+78 +/- 6	---
30	102 +/- 1	177 +/- 3	+75 +/- 2	98
60	103 +/- 1	176 +/- 2	+73 +/- 1	97
120	103 +/- 2	174 +/- 1	+71 +/- 1	96
250	103 +/- 1	171 +/- 2	+68 +/- 2	94
500	103 +/- 1	163 +/- 2	+60 +/- 2	90

a: Initial means group body weights +/- standard error of the mean. Subsequent calculations are based on those animals surviving to the end of the study.

b: Mean body weight change of the survivors of the group +/- standard error of the mean. Final body weights were taken during week 12 of the study.