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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The endpoint conclusion is based on weight-of-evidence: relevant data show that two analogues are not mutagenic in vitro. This data is read-across to the registered substance.

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reason / purpose:
read-across source
Key result
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
See section Additional information on results
Vehicle controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
See section Additional information on results
Vehicle controls validity:
valid
Positive controls validity:
valid
Conclusions:
The mutagenic potential of a related member of MIRANOL ULTRA L32 (amphoacetates C12) was assessed in Salmonella typhimurium bacterial strains and Escherichia coli , according to EC method B.13-B.14 and modified OECD guideline 471, and in compliance with Good Laboratory Practice. It was concluded that the test item is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay. This data is read-across to the registered substance.
Executive summary:

The mutagenic potential of a related member of MIRANOL ULTRA L32 (amphoacetates C12) was assessed in Salmonella typhimurium bacterial strains and Escherichia coli , according to EC method B.13-B.14 and modified OECD guideline 471, and in compliance with Good Laboratory Practice.

The histidine-requiring S. typhimurium mutants TA 1535, TA 1537, TA 98, TA 100 and tryptophan-requiring E.coli mutant WP2uvrA were used in the presence and the absence of metabolic activation (+/- S9 -mix). All determinations were made in triplicate. The first experiment was conducted using a plate-incorporation method at 5 doses levels up to 5000 µg/plate. The second and third experiment were performed using a pre-incubation method. Simultaneously negative (solvent) and positive controls were used in all experiments.

In a preliminary range-finding assay, toxicity was seen.

In the plate-incorporation assay, 4 strains were treated at dose levels 313 to 5000 µg/plate, while TA100 was treated at 156 to 2500 µg/plate. Slight toxicity was observed at the highest dose level with TA1535, TA1537 and TA100, but there was no increase in the revertant numbers in any of the strains tested.

The second assay was performed with a pre-incubation step and using the same dose-range. Moderate thinning of the background lawn was observed at the highest dose level with TA98 tester strain, while marked toxicity was observed with TA1535, TA1537 and TA100 at the 3 highest dose levels. A third experiment was then performed with these 3 strains using a lower dose-range (39.1- 625 µg/plate without S9-mix, and 78.1-1250 µg/plate only for TA1535 and TA1537 in the presence of S9 -mix).

The test material did not cause any two-fold or greater increase in the mean number of revertant colonies in the test plates, both in the absence and the presence of metabolic activation. Positive controls gave the expected increases in the number of revertants, with and without S9 mix.

Under the conditions of this study, the test material did not demonstrate any in vitro mutagenic activity in these bacterial test systems. This data is read-across to the registered substance.

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reason / purpose:
read-across source
Key result
Species / strain:
lymphocytes: human peripheral blood lymphocytes (HPBL)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Conclusions:
A chromosome aberration study with Reaction products of 1H-Imidazole-1-ethanol, 4,5-dihydro-2-(C11 alkyl) derivs. and sodium hydroxide and chloroacetic acid (aqueous solution; amphoacetates C12) was performed according to OECD 473 guideline and GLP principles, in peripheral human lymphocytes in two independent experiments with and without metabolic activation. It is concluded that the substance is not clastogenic in human lymphocytes. This result is read-across to the registered substance.
Executive summary:

In accordance with OECD 473 (1998), EU Method B.10 (2008) and according to GLP principles, a chromosome aberration study with Reaction products of 1H-Imidazole-1-ethanol, 4,5-dihydro-2-(C11 alkyl) derivs. and sodium hydroxide and chloroacetic acid (aqueous solution; amphoacetates C12) was performed in human peripheral blood lymphocytes in two independent experiments with and without metabolic activation. The dosing preparations were adjusted for the surfactant content of the substance and were set based on the observed toxicity in the dose range finding study (no precipitation was observed up to and including the top dose of 5000 µg/mL).

In the first cytogenetic assay, the cells were treated for 4 hr (with and without S9-mix) and for 20 hr (without S9-mix) with the substance at doses ranging from 30 to 250 µg/mL. In the second cytogenetic assay, the cells were treated for 4 hr (with S9 -mix) with the substance at doses ranging from 30 to 200 µg/mL.

Toxicity was reached at the dose levels selected for scoring. In all exposure groups, the percentage of cells with structural or numerical aberrations in the test substance-treated group was not significantly increased relative to solvent control at any dose level (p > 0.05, Fisher's Exact test). The number of cells with chromosome aberrations found in the solvent and positive control cultures was within the laboratory historical control data range.

No effects of the substance on the number of polyploid cells and cells with endoreduplicated chromosomes were observed both in the absence and presence of S9-mix.

Based on the results of this study, it is concluded that the substance does not disturb mitotic processes and cell cycle progression and is not clastogenic in human lymphocytes. This result is read-across to the registered substance.

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reason / purpose:
read-across source
Key result
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Untreated negative controls validity:
valid
Positive controls validity:
valid
Conclusions:
A mouse lymphoma assay was conducted with Alkylamidoamine glycinate, majority in C12 & 14 (amphoacetates C8-C18), in accordance with OECD 476 and according to GLP principles. It was concluded that the substance is not mutagenic in the TK mutation test system. This result is read-across to the registered substance.
Executive summary:

A mouse lymphoma assay was conducted with Alkylamidoamine glycinate, majority in C12 & 14 (amphoacetates C8-C18), in accordance with OECD 476 and according to GLP principles. The spontaneous mutation frequencies in the solvent-treated control cultures were between the minimum and maximum value of the historical control data range. Positive control chemicals, methyl methane sulfonate and cyclophosphamide induced appropriate responses. In the absence of S9-mix, Alkylamidoamine glycinate, majority in C12 & 14 (amphoacetate) did not induce a significant increase in the mutation frequency in the first experiment. This result was confirmed in a repeat experiment with modifications in the duration of treatment time. In the presence of S9-mix, Alkylamidoamine glycinate, majority in C12 & 14 (amphoacetate) did not induce a significant increase in the mutation frequency in the first experiment. This result was confirmed in an independent experiment with modifications of the S9-mix concentration added for metabolic activation. In conclusion, the substance is not mutagenic in the TK mutation test system. This result is read-across to this substance, member of the same chemical category. This result is read-across to the registered substance.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Read-Across: Ames study (OECD 471)

The mutagenic potential of a substance analogue (amphoacetates C12) (aqueous solution) was assessed in Salmonella typhimurium bacterial strains and Escherichia coli, in accordance with OECD 471 and according to GLP principles. The histidine-requiring S. typhimurium mutants TA 1535, TA 1537, TA 98, TA 100 and tryptophan-requiring E.coli mutant WP2uvrA were used in the presence and the absence of metabolic activation. All determinations were made in triplicate. The first experiment was conducted using a plate-incorporation method at 5 dose levels up to 5000 µg/plate (aqueous solution) . The second and third experiments were performed using a pre-incubation method. Simultaneously negative (solvent) and positive controls were used in all experiments. In a preliminary range-finding assay, toxicity was seen. In the plate-incorporation assay, 4 strains were treated at dose levels 313 to 5000 µg/plate, while TA100 was treated at 156 to 2500 µg/plate. Slight toxicity was observed at the highest dose level with TA1535, TA1537 and TA100, but there was no increase in the revertant numbers in any of the strains tested. The second assay was performed with a pre-incubation step and using the same dose-range. Moderate thinning of the background lawn was observed at the highest dose level with TA98 tester strain, while marked toxicity was observed with TA1535, TA1537 and TA100 at the 3 highest dose levels. A third experiment was then performed with these 3 strains using a lower dose-range (39.1- 625 µg/plate without S9-mix, and 78.1-1250 µg/plate only for TA1535 and TA1537 in the presence of S9 -mix). The substance (aqueous solution) did not cause any two-fold or greater increase in the mean number of revertant colonies in the test plates, both in the absence and the presence of metabolic activation. Positive controls gave the expected increases in the number of revertants, with and without S9 mix. Under the conditions of the study, the substance did not demonstrate any in vitro mutagenic activity in these bacterial test systems. This result is read-across to this substance, member of the same chemical category. The read-across from this analogue to the registered substance for this result is considered scientifically justified based on the overall information available (the rationale to read across the data is attached in Section 13

 

Read-Across: Chromosome aberration study (OECD 473)

In accordance with OECD 473 (1998), EU Method B.10 (2008) and according to GLP principles, a chromosome aberration study with a substance analogue (amphoacetates C12) (dissolved in water) was performed in human peripheral blood lymphocytes in two independent experiments with and without metabolic activation. The dosing preparations were adjusted for the surfactant content of the substance and were set based on the observed toxicity in the dose range finding study (no precipitation was observed up to and including the top dose of 5000 µg/mL). In the first cytogenetic assay, the cells were treated for 4 hr (with and without S9-mix) and for 20 hr (without S9-mix) with the substance at doses ranging from 30 to 250 µg/mL. In the second cytogenetic assay, the cells were treated for 4 hr (with S9 -mix) with the substance at doses ranging from 30 to 200 µg/mL. Toxicity was reached at the dose levels selected for scoring. In all exposure groups, the percentage of cells with structural or numerical aberrations in the test substance-treated group was not significantly increased relative to solvent control at any dose level (p > 0.05, Fisher's Exact test). The number of cells with chromosome aberrations found in the solvent and positive control cultures was within the laboratory historical control data range. No effects of the substance on the number of polyploid cells and cells with endoreduplicated chromosomes were observed both in the absence and presence of S9-mix. Based on the results of this study, it is concluded that the substance does not disturb mitotic processes and cell cycle progression and is not clastogenic in human lymphocytes. These results are read-across to the registered substance. The rationale to read across the data is attached in Section 13.

 

Read-Across: in vitro gene mutation assay (OECD 476)

A mouse lymphoma assay was conducted with a substance analogue (amphoacetates C8 -C18) in accordance with OECD 476 and according to GLP principles. In the absence of S9-mix, the substance did not induce a significant increase in the mutation frequency in the first experiment. This result was confirmed in a repeat experiment with modifications in the duration of treatment time. In the presence of S9-mix, the substance did not induce a significant increase in the mutation frequency in the first experiment. This result was confirmed in an independent experiment with modifications of the S9-mix concentration added for metabolic activation. The spontaneous mutation frequencies in the solvent-treated control cultures were between the minimum and maximum value of the historical control data range. Positive control chemicals, methyl methane sulfonate and cyclophosphamide induced appropriate responses. In conclusion, the substance is not mutagenic in the TK mutation test system. This result is read-across to this substance, member of the same chemical category. The rationale to read across the data is attached in Section 13.

Justification for classification or non-classification

Based on the current data-set it is concluded that there are no indications that the registered substance has mutagenic properties and the substance does not need to be classified for mutagenicity in accordance with the CLP Regulation.