Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

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

Genetic toxicity in vivo

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

Additional information

Only limited information on the mutagenic potential has been reported for D,L-alpha-tocopherol per se. Therefore, data

from the structural related Vitamin E ester D,L-alpha-tocopheryl acetate, which will be hydrolzed to free tocopherol under physiological conditions, and the analogue substance gamma-tocopherol are taken into account for the evaluation of a mutagenic potential. Please see read-across justification section 13.

In vitro genotoxicity

D,L-alpha- tocopheryl-acetate was evaluated for mutagenic activity in the Ames test (OECD 471, GLP) in absence and in presence of metabolic activation in five Salmonella typhimurium test strains (TA1535, TA97, TA98, TA100, and TA102). The concentration ranges from 50 to 5000 μg/plate.

No increase in the number of revertant colonies was apparent. It is concluded that D,L-alpha-tocopheryl-acetate is not mutagenic in the Ames test under the described experimental conditions (E.Gocke , 1999).

In another Ames test no increase in the number of revertants and no toxicity was observed with Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA 1537, both in the absence and presence of rat S9 -mix. The dose levels were 20, 100, 500, 2500 and 5000 µg/plate (BASF AG, 1989).

 

D,L-alpha tocopheryl acetate at concentrations between 75 and 1800 μg/ml was tested in an in vitro cytogenetics assay using human lymphocytes (OECD 473, GLP) with and without metabolic activation. After exposure to the test article the frequency of cells with structural or numerical chromosome aberrations was not increased to a biologically relevant extent. It is concluded that D,L-alpha tocopheryl acetate is not clastogenic or aneuploidogenic under the described experimental conditions (A. Chetelat, 1999).

No increase in frequency of chromosome aberrations was observed in a chromosome aberration test with D,L-alpha-tocopherol in Chinese hamster fibroblast cells (CHL), in the absence of metabolic activation. The dose levels were 0.125, 0.25 and 0.5 mg/ml (Ishidate, 1984).

 

In a gene mutation test in Chinese hamster ovary AS52 cells, D-gamma-tocopherol had no effect on mutant frequency in the absence of metabolic activation at dose levels of 2.9 and 14.6 µg/ml. A significant decrease in dosing efficiency was observed at the high concentration when compared to controls; cloning efficiency was approximately 85 -90% (control = 100%). D-gamma-tocopherol was therefore considered to cause no gene mutation under the applied test conditions in this mammalian gene mutation test (Cornwell, 2002).

 

In vivo genotoxicity

The effect of vitamin E (alpha-tocopheryl acetate) on the bone marrow of mice was investigated. The mice were fed a diet containing the test substance at concentrations of 0, 30 or 1000 ppm (corresponding to doses of ca. 0, 6 or 200 mg/kg bw/d, respectively) for up to 50 weeks. Reticulocytes from blood samples collected at various time intervals were used for micronucleus analysis. The incidence of reticulocytes containing micronuclei did not increase in the low group and did not decrease in the high group when compared with the mid group. There were no statistically significant intergroup differences at each time point (Umegaki, 1997).


Short description of key information:
No effects were noted in
- an Ames test, OECD 471, GLP, performed with the structural analogue D,L-alpha-tocopheryl acetate (E.Gocke, 1999)
- an in vitro chromosome aberration test in CHL cells, near-guideline and non-GLP with D,L-alpha-tocopherol (Ishidate, 1984)
- an in vitro chromosome aberration test in human lymphocytes, OECD 473, GLP, performed with the structural analogue D,L-alpha-tocopheryl acetate (A.Chetelat, 1999)
- an in vitro gene mutation test in CHO cells, near guideline and non-GLP with the structural analogue D-gamma-tocopherol (Cornwell, 2002),
- an in vivo micronucleus test, similar to OECD 474, non-GLP, performed with structural analogue D,L-alpha-tocopheryl acetate (Umegaki, 1997).

Thus, vitamin E, including D,L-alpha tocopherol, is considered to be non-genotoxic.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

According to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008 classification is not warranted.