Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The test substance Isosorbid Monooleate is not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay and in the HPRT locus assay under in vitro conditions in CHO cells, in the absence and the presence of metabolic activation. Isosorbid Monooleate is considered not to have a chromosome-damaging (clastogenic) effect nor to induce numerical chromosomal aberrations (aneugenic activity) under in vitro conditions in V79 cells in the absence and the presence of metabolic activation.

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

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Ames test

In a GLP compliant study (BASF SE, 2014) performed according to OECD guideline 471, the test substance Isosorbide Monooleate 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 (TA 1535, TA 100, TA 1537, TA 98) and Escherichia coli (E.coli WP2 uvrA), in a reverse mutation assay. The test substance was tested both in the standard plate test (33 – 7000 µg/plate) and in the pre-incubation test (10 – 3500 µg/plate) both with and without the addition of a metabolizing system (phenobarbital and β-naphthoflavone induced rat liver S9 mix). Precipitation of the test substance was found from about 1000 µg/plate onward with and without S9 mix. A bacteriotoxic effect was occasionally observed depending on the strain and test conditions from about 1000 µg/plate onward. A biologically relevant increase in the number of his+ or trp+ revertants was not observed in the standard plate test or in the pre-incubation test either without S9 mix or after the addition of a metabolizing system. Thus, under the experimental conditions of this study, the test substance Isosorbide Monooleate is not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay in the absence and the presence of metabolic activation.

 

Gene mutation assay HPRT

In a GLP compliant study (BASF SE, 2015) performed according to OECD guideline 476, the substance Isosorbide Monooleate was assessed for its potential to induce gene mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus in Chinese hamster ovary (CHO) cells in vitro. Two independent experiments were carried out, both with and without the addition of liver S9 mix from phenobarbital- and β-naphthoflavone induced rats (exogenous metabolic activation). According to an initial range-finding cytotoxicity test for the determination of the experimental doses and taking into account the cytotoxicity actually found in the main experiments, the following concentrations were tested. The tested concentrations of the 1st main experiment were 3.1, 6.3, 12.5, 25.0, 50.0, and 100.0 μg/mL without metabolic activation and 1.6, 3.1, 6.3, 12.5, 25.0, and 50.0μg/mL with metabolic activation. The test concentrations of the 2nd main experiment were 4.4, 8.8, 17.5, 35.0, 70.0, and 100.0 μg/mL without metabolic activation and 2.2, 4.4, 8.8, 17.5, 35.0, and 70.0 μg/mL with metabolic activation. Following attachment of the cells for 20 - 24 hours, cells were treated with the test substance for 4 hours in the absence and in the presence of metabolic activation. Subsequently, cells were cultured for 6 - 8 days and then selected in 6-thioguanine-containing medium for another week. Finally, the colonies of each test group were fixed with methanol, stained with Giemsa and counted. The vehicle controls gave mutant frequencies within the range expected for the CHO cell line. Both positive control substances led to the expected increase in the frequencies of forward mutations. In this study, in the 1st and 2nd Experiment, at least the highest concentrations evaluated for gene mutations were clearly cytotoxic in the absence and the presence of metabolic activation. Based on the results of the present study, the test substance did not cause any relevant increase in the mutant frequencies without S9 mix or after the addition of a metabolizing system in two experiments performed independently of each other. Thus, under the experimental conditions of this study, the test substance Isosorbide Monooleate is not mutagenic in the HPRT locus assay under in vitro conditions in CHO cells in the absence and the presence of metabolic activation.

 

Micronucleus test

In a GLP compliant study (BASF SE, 2015) performed according to OECD guideline 487, the substance Isosorbide Monooleate was assessed for its potential to induce micronuclei in V79 cells in vitro (clastogenic or aneugenic activity). Two independent experiments were carried out, with and without the addition of liver S9 mix from phenobarbital- and β-naphthoflavone induced rats (exogenous metabolic activation). According to an initial range-finding cytotoxicity test for the determination of the experimental doses, the following concentrations were tested. The cells in the 1st experiment were exposed for 4 hours and harvested after being 24 hours in the medium. The tested concentrations in the 1st experiment were 3.13, 6.25, 12.50, 25.0, 50.0, and 100.0 μg/mL without metabolic activation and 6.25, 12.50, 25.0, 50.0, 100.0, and 200.0 μg/mL with metabolic activation. The cells in the 2nd experiment were exposed for 24 and 4 hours, and harvested after being 24 and 44 hours in the medium, without and with metabolic activation, respectively. The tested concentrations in the 2nd experiment were 6.25, 12.50, 25.0, 50.0, and 100.00 μg/mL without or with metabolic activation. A sample of 1000 cells for each culture was analyzed for micronuclei, i.e. 2000 cells for each test group. The vehicle controls gave frequencies of micronucleated cells within our historical negative control data range for V79 cells. Both positive control substances, ethyl methanesulfonate and cyclophosphamide, led to the expected increase in the number of cells containing micronuclei. Cytotoxicity indicated by clearly reduced cell count or proliferation index was observed at least at the highest applied test substance concentration in all experimental parts of this study. On the basis of the results of the present study, the test substance did not cause and biologically relevant increase in the number of cells containing micronuclei either without S9 mix or after adding a metabolizing system. Thus, under the experimental conditions described, Isosorbide Monooleate is considered not to have a chromosome-damaging (clastogenic) effect nor to induce numerical chromosomal aberrations (aneugenic activity) under in vitro conditions in V79 cells in the absence and the presence of metabolic activation. 

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result the substance is not considered to be classified for genotoxicity under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.