Butanoic acid, 4-amino-4-oxosulfo-, N-coco alkyl derivs., monosodium salts, compds. with triethanolamine

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The test material is considered to be non-mutagenic in vitro micronucleus test, when tested up to precipitating concentrations.

The test substance is not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay.

The test item is considered to be non-mutagenic in the HPRT assay.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In vitro micronucleus test

The test material, suspended in culture medium, was assessed for its potential to induce micronuclei in human lymphocytesin vitroin two independent experiments.

In each experimental group two parallel cultures were analyzed. Per culture at least 1000 binucleated cells were evaluated for cytogenetic damage.

The highest applied concentration in this study (2105 µg/mL of the test item) was chosen with regard to the purity (95%) of the test item and with respect to the current OECD Guideline 487.

Dose selection of the cytogenetic experiment was performed considering the toxicity data and the occurrence of test item precipitation in accordance with OECD Guideline 487.

In the absence and presence of S9 mix, no cytotoxicity was observed up to the highest evaluated concentration, which showed precipitation.

In the absence and presence of S9 mix, no relevant increase in the number of micronucleated cells was observed after treatment with the test item. In Experiment I in the absence of S9 mix, however, two values (1.45 and 1.13%) after treatment with 24.4 and 42.8 µg/mL, respectively, slightly exceeded the historical control data range (0.08 – 1.12% micronucleated cells). In the presence of S9 mix one value (1.35%), after treatment with 74.8 µg/mL, slightly exceeded the historical control data range (0.16 – 1.08% micronucleated cells). None of these values showed a statistical significance.

Appropriate mutagens were used as positive controls. They induced statistically significant increases in cells with micronuclei.

In conclusion, it can be stated that under the experimental conditions reported, the test item did not induce micronuclei as determined by the in vitro micronucleus test in humanlymphocytes.

Therefore, the test material is considered to be non-mutagenic in this in vitro micronucleus test, when tested up to precipitating concentrations. 

Salmonella typhimurium/Escherichia coli reverse mutation assay

The test substance was tested for its mutagenic potential based on the ability to induce point mutations in selected loci of several bacterial strains, i.e. Salmonella typhimurium and Escherichia coli, in a reverse mutation assay.

STRAINS: TA 1535, TA 100, TA 1537, TA 98 and E. coli WP2 uvrA

DOSE RANGE: 1 μg - 5300 μg/plate (SPT), 1 μg - 5300 μg/plate (PIT)

TEST CONDITIONS: Standard plate test (SPT) and preincubation test (PIT) both with and without metabolic activation (liver S9 mix from induced rats).

SOLUBILITY: Precipitation of the test substance was found in the standard plate from about 2650 μg/plate onward with and without S9 mix.

ACCEPTANCE CRITERIA: The experiment was considered valid since the acceptance criteria were met. The results of the negative as well as the positive controls performed in parallel corroborated the validity of this study, since the values fulfilled the acceptance criteria. In this study with and without S9 mix, the number of revertant colonies in the negative controls was within the range of the historical negative control data for each tester strain. In addition, the positive control substances with and without S9 mix induced a significant increase in the number of revertant colonies within the range of the historical positive control data.

TOXICITY: A bacteriotoxic effect was observed depending on the strain and test conditions from about 100 μg/plate onward.

MUTAGENICITY: A relevant increase in the number of his+ or trp+ revertants (factor ≥ 2: TA 100, TA 98 and E.coli WP2 uvrA or factor ≥ 3: TA 1535 and TA 1537) was not observed in the standard plate test or in the preincubation test without S9 mix or after the addition of a metabolizing system.

CONCLUSION: Under the experimental conditions of this study, the test substance is not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay in the absence and the presence of metabolic activation.

HPRT assay:

The study was performed to investigate the potential of the test material to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster.

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

The first main experiment was performed with a treatment period of 4 hours with and without microsomal activation. The second main experiment was performed with a 24 hours treatment period without microsomal activation and a 4 hours treatment period with microsomal activation.

The maximum test item concentration of the pre-experiment (2105.0 µg/mL) was chosen with respect to the current OECD guideline 476 regarding the water content of the test item.

The test item was suspended in culture medium.

The evaluated experimental points and the results are summarised inTable1.

The highest evaluated concentration in experiment I with and without metabolic activation and in experiment II with metabolic activation was limited by precipitation. The highest evaluated concentration in experiment II without metabolic activation was limited by cytotoxicity.

Cytotoxic effect indicated by an adjusted cloning efficiency I below 50% in both cultures was observed in experiment I at 93.75 µg/mL without metabolic activation, and in experiment II at 125.0 µg/mL and above without metabolic activation.

In experiment I the 95% confidence interval was exceeded at 93.75 µg/mL in culture without metabolic activation, and at 15.63 µg/mL in both cultures with metabolic activation. In experiment II without metabolic activation the 95% confidence interval was exceeded at 31.25 µg/mL in culture I, at 62.5 µg/mL in both cultures, at 150.0 µg/mL in culture I, and at 187.5 µg/mL in culture II. In experiment II with metabolic activation the 95% confidence interval was exceeded at 125.0 µg/mL in culture II. These isolated increases were judged as irrelevant as they were not reproduced and there was no dose dependent increase as indicated by a lacking statistical significance.

A t-test evaluating the data of both parallel cultures at these test points showed a significant response in experiment I at 93.75 µg/mL in the absence of metabolic activation. However, since this was the second concentration were precipitation was observed, this concentration will not be taken into the consideration of the judgement of the outcome of this study. In the presence of metabolic activation in experiment I the t-test showed a significant response at 15.63 µg/mL. However, this was the lowest concentration and there was no significant t-test at any other, even higher concentration. In experiment II with metabolic activation the t-test showed a significant response at 125.0 µg/mL. However no statistical significance was observed in the linear regression analysis. Furthermore the relative adjusted cloningefficiencyI at this concentration was below 10%. Therefore this concentration will not be taken into the consideration of the judgement of the outcome of this study.

A linear regression analysis (least squares) was performed to assess a possible dose dependent increase of mutant frequencies. A significant dose dependent trend of the mutation frequency indicated by a probability value of <0.05 was determined in the first culture of experiment I without metabolic activation. As the trend was not reproduced in the parallel culture it is consequently judged as irrelevant fluctuation. All other experimental parts showed no significant dose dependent trend of the mutation frequency.

All in all, no substantial and reproducible dose dependent increase of the mutation frequency was observed in the main experiments.

Appropriate reference mutagens, used as positive controls, induced a distinct increase in mutant colonies and thus, showed 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, the test material is considered to be non-mutagenic in this HPRT assay.

Justification for classification or non-classification

The available studies are considered reliable and suitable for classification purposes under (EC) No. 1272/2008. 

As a result the substance is not considered to be classified for genotoxicity under (EC) No. 1272/2008.