1,1,2,2,3,3,4,4,4-nonafluoro-N-(2-hydroxyethyl)-N-methylbutane-1-sulphonamide

Endpoint summary

Additional information

The test article was tested in the Salmonella typhimurium reverse mutation assay with four histidine-requiring strains of Salmonella typhimurium (IA1535, TA1537, TA100 and TA98) and in the Escherichia coli reverse mutation assay with a tryptophan-requiring strain of Escherichia coli WP2uvrA. The test was performed in two independent experiments in the presence and absence of S9-mix. In the dose range finding test, the test article was tested up to concentrations of 5000 and 4740 micrograms/plate in the strains TA100 and WP2uvrA, respectively in the absence and presence of 5% (v/v) S9-mix. The test article did not precipitate on the plates at this dose level. In tester strain TA100, toxicity was observed at dose levels of 3330 and 5000 micrograms/plate in the absence of S9-mix and at a dose level of 1000 micrograms/plate and upwards already in the presence of S9-mix. In tester strain WP2uvrA, no toxicity was observed at any of the dose levels tested.

In the first mutation assay, the test article was tested up to concentrations of 5000 micrograms/plate in the absence and presence of 5% (v/v) S9-mix in the strains TA1535, TA1537 and TA98. Toxicity was observed in all three tester strains. In the second mutation assay, the test article was tested up to concentrations of 3330 micrograms/plate in tester strain TA1535, TA1537 and TA100 and up to 5000 micrograms/plate in tester strain TA98 and WP2uvrA in the absence and presence of 10% (v/v) S(-mix. Toxicity was observed in all tester strains, except in test strain WP2uvrA in the presence of S9-mix. The test article did not induce a dose-related, two-fold increase in the number of revertant (His+) colonies in each of the four tester strains and in the number of revertant (Trp+) colonies in tester strain WP2uvrA both in the absence and presence of S9-metabolic activation. These results were confirmed in an independently repeated experiment. Based on the results of this study, it is concluded that the test article is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.

 

The effect of the test article on the number of chromosome aberrations in cultured peripheral human lymphocytes in the presence and absence of a metabolic activation system was studied. The possible clastogenicity of the test article was tested in two independent experiments. In the first cytogenic assay, the test article was tested up to 420 micrograms/ml for a three hour exposure time with a 24 hour fixation time in the absence and presence of S9-mix. Appropriate toxicity was reached at this dose level. In the second cytogenic assay, the test article was tested up to 100 micrograms/ml for a 24 hour continuous exposure time with a 48 hour fixation time in the absence of S9-mix. Appropriate toxicity was reached at these dose levels. In the presence of 1.8 (v/v) S9-fraction, the test article was tested up to 450 micrograms/ml for a 3h exposure time with a 48 h fixation time. The test article precipitated in the culture medium at this dose level. Positive control chemicals, mitomycin C and cyclophosphamide, both produced a statistically significant increase in the incidence of cells with chromosome aberrations, indicating that the test conditions were adequate and that the metabolic activation system (9-mix) functioned properly. The test article did not induce a statistically significant or biologically relevant increase in the number of cells with chromosome aberrations in the absence and in the presence of S9-mix, in two independently repeated experiments. It is concluded that this test is valid and that the test article is not clastogenic in human lymphocytes under the experimental conditions described in this report.

The test substancewas evaluated for itsability to induce forward mutations at the thymidine kinase locus in L5178Y mouse lymphoma cellsin the presence and absence of an exogenous metabolic activation system. The study was conducted according to OECD 490 (2016) in compliance with OECD GLP regulations. Dimethyl sulfoxide (DMSO) was used as the vehicle. In the preliminary toxicity assay, the concentrations tested were 7.81, 15.6, 31.3, 62.5, 125, 250, 500, 1000 and 2000 µg/mL. The maximum concentration evaluated approximated the limit dose for this assay. Visible precipitate was observed at concentrations ≥500 µg/mL at the beginning and end of treatment. Relative suspension growth (RSG) was 67, 93 and 25% at concentrations of 125 µg/mL with 4-hour treatment with and without S9 and 24-hour treatment without S9, respectively. RSG was 0% at all higher concentrations using all treatment conditions. Based upon these results, the concentrations chosen for the definitive mutagenicity assay were 75, 100, 125, 150, 175, 200, 225 and 250 µg/mL (4-hour treatment with and without S9) and 25, 50, 75, 100, 125, 150, 175 and 200 µg/mL (24-hour treatment without S9). In the initial definitive mutagenicity assay (B1), no visible precipitate was observed at the beginning or end of treatment. Cultures treated at concentrations of 75, 100, 125, 150, 175 and 200 µg/mL (4-hour treatment with S9) (one replicate at 125 µg/mL was lost due to dilution error and one plate at 200 µg/mL was not cloned due to excessive toxicity), 75, 100, 125, 150 and 175 µg/mL (4-hour treatment without S9) and 25, 50, 75, 100 and 125 µg/mL (24-hour treatment without S9) exhibited 8 to 69%, 16 to 97% and 10 to 89% RSG, respectively, and were cloned. Relative total growth of the cloned cultures ranged from 12 to 69% (4‑hour treatment with S9), 18 to 91% (4-hour treatment without S9) and 9 to 76% (24‑hour treatment without S9). The cloning efficiency of the vehicle control for 4-hour treatment without S9 was above 120%; therefore, this condition was repeated. No increases in induced mutant frequency ≥90 mutants/10⁶clonable cells were observed under 4‑hour treatment with S9 and 24‑hour treatment without S9.  Based upon the above results, the concentrations chosen for the initial repeat of definitive mutagenicity assay (B2) were 75, 125, 150, 175, 180, 185, 200 and 225 µg/mL (4-hour treatment without S9). No visible precipitate was observed at the beginning or end of treatment. Four concentrations of test substance were not available for cloning and evaluation, therefore this condition was repeated with different concentrations. Based upon the above results, the concentrations chosen for the final repeat of definitive mutagenicity assay (B3) were 37.5, 75, 125, 150, 160, 175, 180, 185 and 200 µg/mL (4-hour treatment without S9). No visible precipitate was observed at the beginning or end of treatment. Cultures treated at concentrations of 75, 125, 150, 160, 175 µg/mL exhibited 11 to 124%, RSG, respectively, and were cloned. Relative total growth of the cloned cultures ranged 10 to 112%. No increases in induced mutant frequency ≥90 mutants/10⁶clonable cells were observed under any treatment condition. A dose dependent trend was observed.  However, it should be noted that the IMF was below 90 mutants/10⁶clonable cells.  Therefore, the trend was considered to be biologically irrelevant. Based on the results of the study, the test article does not induce forward mutations at the thymidine kinase locus in L5178Y mouse lymphoma cells, in the presence and absence of metabolic activation (S9).

Short description of key information:
In vitro genetic toxicity studies were conducted on C4 Alcohol. The results of the studies are:

Bacterial Reverse Mutation Assay: Negative when tested according to OECD 471.

Chromosome Aberration: Negative when tested according to OECD 473.

Mouse Lymphoma Assay: Negative when tested according to OECD 490.

Endpoint Conclusion:

Criteria for classifying the test article as mutagenic are not met.

Justification for classification or non-classification

Criteria for classifying the test article as mutagenic are not met.