cis-4-tert-butylcyclohexyl acetate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

- Ames test: non mutagenic (WoE)

- MLA: non mutagenic (read-across, OECD 476, GLP, K, rel. 2)

- CAT: non clastogenic (read-across, OECD 473, K, rel.2)

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Table 7.6/1: Summary of genotoxicity tests

 

Test n°	Test substance	Test / Guideline Reliability	Focus	Strains tested	Metabolic activation	Test concentration (µg/plate/mL)	Statement
1   Envigo, 2013	Cis-4-tert-butylcyclohexyl acetate	Ames Test (OECD 471) K, rel. 1	Gene mutation	TA 1535, TA 1537 TA 98 TA 100 WP2 uvrA	-S9 +S9	Up to 5000	-S9 : non mutagenic +S9 : non mutagenic
2   SITEK, 2009a	2-tert-pentylcyclohexyl acetate	Ames Test (OECD 471) S, rel. 2	Gene mutation	TA 1535, TA 1537 TA 98 TA 100	-S9 +S9	Up to 5000 µg/plate or up to cytotoxic concentration	-S9 : non mutagenic +S9 : non mutagenic
3   SITEK, 2009b	2-tert-pentylcyclohexyl acetate	ML/TK test (OECD 476) K, rel. 2	Gene mutation	mouse lymphoma L5178Y cells	-S9 +S9	-S9: up to 26 µg/mL +S9: up to 115 µg/mL	-S9 : non mutagenic +S9 : non mutagenic
4   SITEK, 2009c	2-tert-pentylcyclohexyl acetate	CHO CAT (OECD 473) K, rel. 2	Chromosomal aberration	Chinese Hamster Ovary cells	-S9 +S9	-S9: up to 45 µg/mL +S9: up to 90 µg/mL	-S9 : non clastogenic +S9 : non clastogenic

 

Gene mutation Assays (Tests n° 1 -3):

A bacterial reverse mutation assay (Ames test) was performed with cis-4-tert-butylcyclohexyl acetate (Test n°1). No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains in any tests, with any dose of test materials, either in the presence or absence of metabolic activation. Cis-4-tert-butylcyclohexyl acetate did not induce gene mutations in bacteria whereas all positive control chemicals (with and without metabolic activation) induced significant increase of colonies. Cis-4-tert butylcyclohexyl acetate is therefore considered as non-mutagenic according to the Ames test. Test n°2, conducted on the source substance, 2-tert-pentylcyclohexyl acetate (containing approximately 15% of 4-tert butylcyclohexyl acetate (cis and trans isomers)), supports the conclusion that the two substances have the same genotoxicity profile, and therefore confirms that results obtained form 2-tert-pentylcyclohexyl acetate studies can be extended to 4-tert-butylcyclohexyl acetate (Cf. section 13 for read-across justification).

No mammalian gene mutation test was located on 4-tert-butylcylclohexyl acetate. However, inability to produce gene mutation was confirmed on the source substance, 2-tert-pentylcyclohexyl acetate using an in vitro forward mutation assay in mouse lymphoma TK L5178Y cells (ML/TK test) (Test n°3). None of the dose levels up to the cytotoxicity limit with 2-tert-pentylcyclohexyl acetate, either in the presence or absence of metabolic activation, induced significant mutant frequency increases in the initial or repeat tests. 2-tert-pentylcyclohexyl acetate does not induce forward mutations at the TK locus in L5178Y mouse lymphoma cells under activation and non-activation conditions whereas both positive control chemicals (with and without metabolic activation) induced significant mutant frequency increases. 2-tert-pentylcyclohexyl acetate, and by analogy cis-4-tert butylcyclohexyl acetate, are therefore considered as negative for inducing forward mutations at the TK locus in L5178Y mouse lymphoma cells under activation and non-activation conditions used in this assay. This result confirms the results of the Ames tests and extends the non-mutagenic effect of cis-4-tert-butylcyclohexyl acetate to mammalian cells.

Chromosomal aberration (Test n°4):

No mammalian chromosome aberration test was located on cis-4-tert-butylcylclohexyl acetate. However, the clastogenic potential of the source substance, 2-tert-pentylcylclohexyl acetate, was determined using an in vitro chromosome aberration test in Chinese hamster ovary cells, which measures the potential of a substance to increase the incidence the of structural chromosome aberrations in cultured Chinese hamster ovary cells. None of the dose levels up to the cytotoxicity limit with 2-tert-pentylcylclohexyl acetate, either in the presence or absence of metabolic activation, induced significant increases in the frequency of cells with aberrations in either of two experiments. 2-tert-pentylcylclohexyl acetate does not induce structural aberrations in the chromosomes of Chinese hamster ovary cells under activation and non-activation conditions, whereas both positive control chemicals (with and without metabolic activation) induced significant increases in the frequency of aberrant cells. 2-tert-pentylcylclohexyl acetate, and by analogy cis-4-tert-butylcylclohexyl acetate, are therefore considered as negative for inducing chromosomal mutations in Chinese hamster ovary cells under activation and non-activation conditions used in this assay.

Justification for classification or non-classification

Harmonised classification:

The substance has no harmonised classification according to the Regulation (EC) No. 1272/2008.

Self-classification:

Based on the available information, no self-classification is proposed.