Isostearic acid, esters with methyl α-D-glucoside

Endpoint summary

Additional information

Bacterial reverse gene mutation assay

In a reverse gene mutation assay in bacteria according to OECD guideline 471, strains of S. typhimurium (TA 1535, TA 1537, TA 98, and TA 100) and E. coli (WP2 uvrA) were exposed to Isostearic acid, esters with methyl α-D-glucoside at concentrations of 33, 100, 333, 1000, and 3330 µg/plate (dose range finding test: 3, 10, 33, 100, 333, 1000, 3330, and 5000 µg/plate) in the presence and absence of mammalian metabolic activation as plate incorporation assay.

All bacterial strains showed negative responses over the entire dose range, i.e. no significant dose-related increase in the number of revertants in two independently repeated experiments. The negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly. There was no evidence of induced mutant colonies over background, and no biologically relevant decrease in the number of revertants at any of the concentrations tested in all tester strains in the absence and presence of S9-mix.

Based on the results of this study it is concluded that Isostearic acid, esters with methyl α-D-glucoside is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.

This study is classified asacceptable. It satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) dat

Mammalian cell cytogenetic assay

In a mammalian cell cytogenetic assay, human peripheral blood lymphocyte cultures were exposed to Isostearic acid, esters with methyl α-D-glucoside in dimethyl sulfoxide at concentrations of 0, 33, 100, and 333 µg/ml culture medium (3 h exposure time, 24 h fixation time with and without metabolic activation in the first cytogenetic assay). In the second cytogenetic assay concentrations of 0, 100, 300, 400, 500, 600, 700, and 800 µg/ml (24 h and 48 h exposure time and fixation time) without S9-mix and 0, 33, 100, and 300 µg/ml (3 h exposure time, 48 h fixation time) with S9-mix were tested, respectively.

Isostearic acid, esters with methyl α-D-glucoside was tested up to cytotoxic and precipitating concentrations. In the first cytogenetic assay, Isostearic acid, esters with methyl α-D-glucoside was tested up to 333 µg/ml for a 3 h exposure time with a 24 h fixation time in the absence and presence of 1.8% (v/v) S9-fraction. Isostearic acid, esters with methyl α-D-glucoside precipitated in the culture medium at this dose level. In the second cytogenetic assay, Isostearic acid, esters with methyl α-D-glucoside was tested up to 800 µg/ml for 24 h and 48 h continuous exposure time with a 24 h and 48 h fixation time in the absence of S9-mix. Appropriate toxicity was reached at 600 µg/ml. In the presence of S9-mix Isostearic acid, esters with methyl α-D-glucoside was tested up to 300 µg/ml for a 3 h exposure time with a 48 h fixation time. Isostearic acid, esters with methyl α-D-glucoside precipitated in the culture medium at this dose level.

Isostearic acid, esters with methyl α-D-glucoside did not induce a statistically significant or biologically relevant increase in the number of cells with chromosome aberrations in the absence and presence of S9-mix, in either of the two independently repeated experiments.

No biologically relevant effects of Isostearic acid, esters with methyl α-D-glucoside on the number of polyploid cells and cells with endoreduplicated chromosomes were observed, both in the absence and presence of S9-mix. Therefore it can be concluded that Isostearic acid, esters with methyl α-D-glucoside does not disturb mitotic processes and cell cycle progression and does not induce numerical chromosome aberrations under the experimental conditions described in this report.

It is concluded that this test is valid and that Isostearic acid, esters with methyl α-D-glucoside is not clastogenic in human Iymphocytes under the experimental conditions described in this report.

Positive controls induced the appropriate response. There was no evidence of chromosome aberrations induced over background. The number of cells with chromosome aberrations found in the solvent control cultures was within the laboratory historical control data range.

This study is classified as acceptable. This study satisfies the requirement for OECD Test Guideline 473.

 

Mammalian cell gene mutation assay

In a mammalian cell gene mutation assay, mouse lymphoma L5178Y cells cultured in vitro were exposed to Isostearic acid, esters with methyl α-D-glucoside in DMSO. The test was performed in two independent experiments in the absence and presence of S9-mix.

In the first experiment Isostearic acid, esters with methyl α-D-glucoside was tested up to concentrations of 240 µg/ml in the absence of 8% (v/v) S9-mix and up to concentrations of 350 µg/ml in presence of 8% (v/v) S9-mix with an incubation time of 3 hours. Isostearic acid, esters with methyl α-D-glucoside was tested up to cytotoxic levels of 12 and 14% in the absence and presence of S9-mix, respectively.

In the second experiment Isostearic acid, esters with methyl α-D-glucoside was tested up to concentrations of 210 µg/ml in the absence of 12% (v/v) S9-mix and up to concentrations of 375 µg/ml in the presence of 12 (v/v) S9-mix. The incubation times were 24 hours and 3 hours for incubations in the absence and presence of S9-mix, respectively. Isostearic acid, esters with methyl α-D-glucoside was tested up to cytotoxic levels of 19 and 41% in the absence and presence of S9-mix, respectively.

In the absence of S9-mix, Isostearic acid, esters with methyl α-D-glucoside did not induce a significant increase in the mutation frequency in the first experiment. This result was confirmed in an independent repeat experiment with modifications in the duration of treatment time.

In the presence of S9-mix, Isostearic acid, esters with methyl α-D-glucoside did not induce a significant increase in the mutation frequency in the first experiment. This result was confirmed in an independent repeat experiment with modifications in the concentration of the S9-mix for metabolic activation.

Isostearic acid, esters with methyl α-D-glucoside is not mutagenic in the mouse lymphoma L5178Y test system under the experimental conditions described in this report.

The positive controls induced the appropriate response. The spontaneous mutation frequencies in the solvent-treated control cultures were between the minimum and maximum value of the historical control data range.

This study is classified as acceptable. This study satisfies the requirement for Test Guideline OECD 476 for in vitro mutagenicity data.

Short description of key information:
Gene mutagenicity in bacteria (Reverse Mutation Assay)
Ames test (Salmonella typh. strains TA98, TA100, TA1535, TA1537, E. coli WP2 uvrA) with and without metabolic activation: negative
OECD TG 471

Chromosome aberrations in Peripheral Human Lymphocyte Cultures, with and without metabolic activation: negative
OECD TG 473

In-vitro Mammalian Cell Gene Mutation Test (Mouse Lymphoma L5178Y TK+/- cells), with and without metabolic activation: negative
OECD TG 476

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

There were no indications of mutagenic activity in the Bacterial Reverse Mutation Assay, of clastogenicity in-vitro in cultured Peripheral Human Lymphocyte Cultures and of mutagenicity in the mammalian cell culture (Mouse Lymphoma) test system. Therefore according to Directive 67/548/EEC as well as GHS Regulation EC No 1272/2008 no classification and labelling for mutagenic toxicity is necessary.