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Key value for chemical safety assessment

Additional information

Ames Test:

The potential of the test item to induce gene mutations was investigated in the bacterial reverse mutation assay according to guideline OECD 471, EU method B 13/14, and EPA OPPTS 870.5100. The Salmonella typhimurium tester strains TA 98, TA 100, TA 1535, TA 1537, and TA 102 were tested in the plate incorporation test (experiment I) and the pre-incubation test (experiment II) for mutagenic activity. Each assay was conducted with and without metabolic activation (S9 mix from phenobarbital/ß-naphthoflavone induced rats). The test item was dissolved in DMSO. The concentrations, including the controls, were tested in triplicate. The following concentrations were prepared and used in the experiments:

Eperiment I and Experiment II:

31.6, 100, 316, 1000, 2500, and 5000 µg/plate

No precipitation of the test item was observed in any tester strain used in experiment I and II (with and without metabolic activation).

No toxic effects of the test item were noted in any of the five tester strains used up to the highest dose group evaluated (with and without metabolic activation) in experiment I and II.

No biologically relevant increases on revertant colony numbers of any of the five tester strains were observed following treatment with the test item at any concentration level, neither in the present nor absence of metabolic activation in experiment I and experiment II.

The reference mutagens induced a distinct increase of revertant colonies indicating the validity of the experiments.

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used.Therefore, the test item is considered to be non-mutagenic in this bacterial revrse mutation assay.

 

Chromosome aberration test:

The possible potential of the test item to induce structural chromosome aberrations was tested in the Chinese hamster V79 cells in the vitro mammalian chromosome aberration test according to OECD guideline 473, EU method B 10, and EPA OPPTS 870.5375.

The chromosomes were prepared 20 h after start of treatment with the test item. The treatment interval was 4 h with and without metabolic activation (S 9 mix from phenobarbital/ß-naphthoflavone induced rats) in experiment I. In experiment II, the treatment interval was 4 h with and 20 h without metabolic activation. Duplicate cultures were treated at each concentration. 100 metaphases per culture were scored for structural chromosomal aberrations.The following concentrations were evaluated for the microscopic analysis of chromosomal aberrations:

Experiment I:

with and without metabolic activation: 100, 2500 and 5000 µg/mL

Experiment II:

with metabolic activation: 3000, 4000 and 5000 µg/mL

without metabolic activation: 1000, 2500 and 5000 µg/mL

No precipitation and no toxic effects of the test item were noted with and without metabolic activation in all dose groups evaluated in experiment I and II.

In both experiments no biologically relevant increase of the aberration rates was noted after treatment with the test item with and without metabolic activation. The aberration rates of all dose groups treated with the test item were within the historical control data of the negative control.

In the experiments I and II with and without metabolic activation no biologically relevant increase in the frequencies of polyploid cells was found after treatment with the test item as compared to the controls.

EMS (600 and 900 µg/mL) and CPA (0.83 µg/mL) were used as positive controls and induced distinct and biologically relevant increases in cells with structural chromosomal aberrations.

In conclusion, it can be stated that during the described in vitro chromosomal aberration test and under the experimental conditions reported, the test item did not induce structural chromosomal aberrations in the Chinese hamster V79 cell line.Therefore, the test item is considered to be non-clastogenic.

 

HPRT test:

An in vitro cell gene mutation test according to OECD guideline 476 and GLP was conducted with D-Glucose, ether with glycerol using Chinese hamster ovary cells (CHO).

Cells were incubated for 4 hours with metabolic activation and for 4 and 24 hours without metabolic activation. Concentrations of 625, 1250, 2500 and the limit dose of 5000 µg/mL were analyzed for gene mutation. No precipitation or cytotoxicity of about or below 20% of the respective negative control was observed.

The negative controls gave mutant frequencies within the range expected for the CHO cell line. Both positive control substances, Ethylmethanesulfonate and 7,12-Dimethylbenzanthracene, without and with metabolic activation, respectively, led to the expected increase in the frequencies of forward mutations.

Thus, under the experimental conditions of this study, the test substance D-Glucose, ether with glycerol is not mutagenic in the HPRT locus assay under in vitro conditions in CHO cells in the absence and presence of metabolic activation.

In conclusion, D-Glucose, ether with glycerol was found not to be mutagenic in vitro in a bacterial reverse mutation assay and in a chromosomal aberration and a gene mutation test in mammalian cell culture.


Short description of key information:
Ames Test:
The test item is considered to be non-mutagenic in the bacterial reverse mutation assay.

Chromosome aberration test:
Under the experimental conditions, the test item did not induce chromosomal aberrations in vitro.

HPRT test:
Under the experimental conditions, the test item did not induce gene mutation at the HPRT locus.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

No classification and labelling is required according to Regulation No (EC) 1272/2008 (CLP) or Directive 67/548/EEC (DSD) criteria.