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Key value for chemical safety assessment

Additional information

There are reliable in vitro studies available to assess the potential of the test substance for gene mutations in bacteria and in mammalian cells as well as for cytogenicity in mammalian cells.

 

Gene mutation in bacteria:

In a GLP conform study according to OECD guideline 471 the potential of the test substance to induce gene mutations based on the ability to induce back mutations was carried out using the Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 (BASF AG, 1991).

The standard plate test and preincubation test were performed both with and without liver microsomal activation (Aroclor induced rat liver S-9 mix). Each concentration, including the controls, was tested in triplicate. The test item was tested at the following concentrations: 20, 100, 500, 2500 and 5000 µg/plate dissolved in DMSO.

A weakly bacteriotoxic effect was occasionally observed depending on the strain and test conditions at doses ≥ 2500 µg/plate.

An increase in the number of his+ revertants was not observed both in the standard plate test and in the preincubation test either without S-9 mix or after the addition of a metabolizing system.

According to the results of the present study, the test substance Oxooel 13 is not mutagenic in the Ames test under the experimental conditions chosen here.

 

Gene mutation in mammalian cells:

A GLP conform study was performed to investigate the potential of Oxooel 13 to induce gene mutations at the HPRT locus in V79 cells according to OECD guideline 476 (BASF SE, 2010).

The highest concentration (4257 µg/mL) applied in the pre-experiment was limited by the solubility properties of the test item in organic solvents and aqueous medium. The concentration range of the main experiments was limited by cytotoxic effects and phase separation of the test item.

The assay was performed in two independent experiments, using two parallel cultures each.

In the first experiment the treatment period was 4 hours with and without metabolic activation. The second experiment was performed with a treatment time of 4 hours with and 24 hours without metabolic activation.

No substantial and reproducible dose dependent increase of the mutation frequency was observed in both main experiments.

Appropriate reference mutagens, used as positive controls, induced a distinct increase in mutant colonies and thus, confirmed the sensitivity of the test system and the activity of the metabolic activation system.

In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells.

Therefore, Oxooel 13 is considered to be non-mutagenic in this HPRT assay.

 

Cytogenicity in mammalian cells:

In an in vitro micronucleus assay in V79 cells, performed according to GLP and OECD 487 (2010), V79 cells were treated with the test item (99.8%) without metabolic activation for 4 hours at concentrations of 0.78; 1.56; 3.13; 6.25; 12.50; 25.00; 50.00; 100.00 μg/mL, for 24 hours at concentrations of 0.20; 0.39; 0.78; 1.56; 3.13; 6.25; 12.50; 25.00 μg/mL and with metabolic activation for 4 hours at concentration of 39.06; 78.13; 156.25; 312.50; 625.00; 1250.00; 2500.00, 4095.00 μg/mL in one experiment and 12.50; 25.00; 50.00; 100.00, 200.00 μg/mL in a further experiment (BASF SE 2011). The cells were harvested at 24 hours after start of the treatment. The test substance was added pure or diluted with the vehicle acetone to the culture medium in a concentration of 0.5% (v/v). A sample of at least 1000 cells for each culture were analyzed for micronuclei, i.e. 2000 cells for each test group.

The vehicle controls gave frequencies of micronucleated cells within the historical negative control data range for V79 cells. Both positive control substances, ethyl methanesulfonate and cyclophosphamide, led to the expected increase in the number of cells containing micronuclei.

On the basis of the results of the study, the test substance did not cause any relevant increase in the number of cells containing micronuclei either without S9 mix or after adding a metabolizing system in three experiments carried out independently of each other.

Thus, under the experimental conditions described, Oxooel 13 is considered neither to have a chromosome-damaging (clastogenic) effect nor to induce numerical chromosomal aberrations (aneugenic activity) under in vitro conditions in V79 cells in the absence and the presence of metabolic activation.

In conclusion, Oxooel 13 is considered to be non-clastogenic in this V79 cell micronucleus assay.

 


Short description of key information:
in vitro:

Gene mutation in bacteria:
Ames test, S. typhimurium, with and without metabolic activation: negative (GLP, OECD 471, BASF AG, 1991)

Gene mutation in mammalian cells:
HPRT test, V79 cells, with and without metabolic activation: negative (GLP, OECD 476, BASF SE, 2010)

Cytogenicity in mammalian cells:
Micronucleus test, V79 cells, with and without metabolic activation: negative (GLP, OECD 487; BASF SE 2011)

in vivo:
no data available

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC):

The available studies are considered reliable and suitable for classification purposes under 67/548/EEC. As a result the substance is not considered to be classified as a mutagen under Directive 67/548/EEC, as amended for the 28th time in Directive 2001/59/EC.

 

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008:

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified as a mutagen under Regulation (EC) No. 1272/2008.