Adiponitrile

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1984 to 1985

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Outlined protocol was followed.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)

GLP compliance:

yes (incl. QA statement)

Type of assay:

chromosome aberration assay

Test material

Test material information

Constituent 1

Reference substance name:

Adiponitrile

EC Number:

203-896-3

EC Name:

Adiponitrile

Cas Number:

111-69-3

Molecular formula:

C6H8N2

IUPAC Name:

hexanedinitrile

Details on test material:

- Name of test material (as cited in study report): Adiponitrile
- Physical state: Colorless to pale yellow liquid
- Analytical purity: 98%

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

83 male and 83 female albino rats approximately 46-51 days old received from Charles River Breeding Laboratories Inc., Kingston, New York. Animals were acclimated to laboratory conditions for 12 days prior to initiation of the study, housed individually in wire-mesh cages with food and water available ad libitum. 12 hours light and 12 hours dark diurnal cycle was maintained. Relative humidity and temperature were monitored daily.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

Isotonic saline for initial dosing. Follow-up dosing with corn oil.

Details on exposure:

A single dose of the test material was administered by oral gavage to two groups of 15 male and 15 female rats at levels of 0 and 300 mg/kg of body weight.

Frequency of treatment:

Single

Doses / concentrations

Remarks:

Doses / Concentrations:
300 mg/kg
Basis:

No. of animals per sex per dose:

15 female 15 male corn oil
5 female 5 male Cyclophosphamide (40 mg/kg)
15 female 15 male Adiponitrile (300mg/kg)

Examinations

Tissues and cell types examined:

Bone marrow cells.

Details of tissue and slide preparation:

Cells processed according to modified techniques described by Evans (1976) and Killian, et al. (1977). Collected from both gemurs of each animal by aspiration into 5 ml of Hank's Balanced Salt Solution prewarmed to 37°C. Aspirate centrifuged for 5 minutes at 11 rpm, supernate decanted and 5.0 ml of prewarmed 37°C 0.075M KCl was added. After 25 minutes, five drops of freshly prepared fixative was added. Tubes immediately capped, gently inverted and centrifuged for 5 minutes at 1100 rpm. Supernate was decanted and 5 ml of fixative were added slowly down the sides of the rube. Tubes resuspended and recentrifuged for 5 minutes. Supernate was discarded and new addition of 5 ml of fixative was added, cells washe once more with fixative then refridgerated. Chilled preparation centrifuded at 1100 rpm for 5 minutes, supernate decanted, suspended in 0.5-2ml fixative. Final suspension dispersed onto precleaned glass microscope sildes and air-dried. 2-4 slides were made for each animal and marked.

Evaluation criteria:

Abberations characterized:
Chromatid breaks
chromosome breaks
Chromatid and chromosome gaps.
Exchanges
cells with >= 10 aberrations
pulverized cells

Statistics:

The mean mitotic indices, mean chromosome numbers, percent aberrant cells and the mean number of aberrations per cell for each group were statistically compared using the Kruskal-Wallis nonparametric analysis of variance and nonparametric pairwise group comparisions (KW-ANOVA). Body weight data was analyzed by analysis of covariance (ANCOVA). All tests were one-tailed at the 95% confidence interval.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Negative controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative
Under the conditions of the study, Adiponitrile is considered not to be clastogenic.

Executive summary:

The acute oral administration of 300 mg/kg body weight, of Adiponitrile to male and female rats produced no significant increase in the frequency of chromosomal aberrations. The test material produced severe toxicity i n male rats as evidenced by abnormal clinical observations, significant loss of body weight and one animal death. Slight toxicity was seen in female rats as evidenced by

abnormal clinical observations. No significant differences were observed between the vehicle controls and test groups when comparing chromosome number and mitotic indices. Therefore, under the conditions of this study, Adiponitrile is considered not to be clastogenic.