1-(2-Ethylbutyl)cyclohexanecarbonitrile

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

other information

Study period:

04.08.2008 to 11.08.2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The test was performed in accordance with OECD Guideline 471 for the Testing of Chemicals (Bacterial Reverse Mutation Test. Adopted 21st July 1997) and the test Method BI 3/B14 of Commission Directive 2000/32/EC. The study was performed according to the GLPs.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

CAT-Nitrile

IUPAC Name:

CAT-Nitrile

Method

Target gene:

Characteristics of the five strains are described in the following table:
TA98 His D 3052 uvrB pKM101 rfa Frameshift
TA I O0 His G 46 uv rB pKM 1 O1 rfa Base-pair substitution
TA 102 His G 428 pKM 1 O1 - Base- pa¡ r substitut ion
TA 1535 His G 46 UVrB - rfa Base-pair substitution
TA1537 His C 3076 WrB - rfa Frameshift

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

5-1.67-0.5-0.167 and 0.05 mg/plate

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

The controls of the test were in concordance with the expected results: - Sterility test showed no contamination during the study. - No cytotoxic effect was observed. - All positive controls performed showed valid ratios (R) above 2.5. - Positive and negative controls showed absolute numbers of revertant colonies comparable to historical data. - No concentration of the test item showed a biological significant increase (R equal or greather than 2.5) of the number of revertant either with or without S9 metabolic activation. - No dose response was observed in none of the tested bacterial strains.

Applicant's summary and conclusion

Conclusions:

The following conclusions can be inferred from the obtained results:
- No test substance showed ratios (R) above 2.5 as compared to the negative control, either with or without S9 metabolic activation.
- No dose response was observed in none of the tested bacterial strains.
Based on the results obtained in this study, the test item CAT Nitril was found to be NON MUTAGENIC and NON-PROMUT AGENIC under the test conditions.

Executive summary:

The present Bacterial Reverse Mutation Test (Ames test) was performed in order to evaluate the mutagenic potential of the test item. The test was performed in accordance with OECD Guideline 471 for the Testing of Chemicals (Bacterial Reverse Mutation Test. Adopted 21 st July 1997) and the test Method B13/B14 of Commission Directive 2000/32/EC. Doses ranging from 5 microL to 0.05 microL per plate were tested, where the maximum concentration used was 5mg/plate. No cytotoxicity was observed at any dose. Suspensions of amino-acid reqUiring strains of Salmonella typhimurium (TA98, TA100, TA102, TA1535, TA1537) were exposed by the direct plate incorporation method to five doses of the test item in the presence and in the absence of an exogenous metabolic activation system. Both tests were repeated with the pre-incubation method. The bacteria were allowed to grow for approximately 72h. Only revertant bacteria due to point or frameshift-mutations at specific locus are able to grow, forming colonies. These colonies were counted and compared to the number of spontaneous revertant colonies on solvent control plates (negative control). Similarly, specific standard mutagens were tested and used as positive controls. Based on the results obtained in this study, the test item CAT Nitril was found to be NON MUTAGENIC and NON-PROMUTAGENIC under the test conditions.