coconut oil, reaction products with glycerol esters of 3,5-bis(1,1-dimethylethyl)-4-hydroxybenzenepropanoic acid

Endpoint summary

Additional information

In vitro:

In a reverse gene mutation assay in bacteria (RCC 570401, 1996), performed according to OECD guideline 471, S. typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvr A were exposed to the test substance (dissolved in DMSO) at concentrations of 33.3, 100, 333.3, 1000, 2500 and 5000 µg/plate in the presence and absence of mammalian metabolic activation (S9 liver microsomal fraction from phenobarbital treated rats). In the positive control groups (sodium azide, 4-nitro-o-phenylene-diamine, methyl methane sulfonate, 2-aminoanthracene) a clear increase in the number of revertants was observed (at least 13-fold). There was no evidence of induced mutant colonies over background or dose related increase in the test groups.

In a mammalian cell cytogenetics assay (chromosomal aberration; RCC 570402, 1996), performed according to OECD guideline 473,

V79 cell cultures were exposed to the test substance (dissolved in DMSO) at concentrations of 30 up to 5000 µg/ml in the presence and absence of mammalian metabolic activation (liver S9-mix from Phenobarbital/b-Naphthoflavone treated rats). Positive controls (cyclophosphamide, ethylmethanesulphonate) induced the appropriate response. In the treated groups, some aberrations were noted but no statistically significant increases in the frequency of aberrations per cell or in the proportion of aberrant metaphases at any dose level evaluated with or without S9 Mix were observed. Hence, treatment of V79 cells with the test substance revealed an ambiguous result concerning the induction of chromosomal aberrations.

In vivo:

In a NMRI mouse bone marrow micronucleus assay (RCC 582100, 1997), performed according to OECD guideline 474, 5 male and 5 female animals per dose were treated once by gavage with the test substance, which was dissolved in polyethylene glycol, at doses of 200, 670, and 2000 mg/kg bw. Bone marrow cells were harvested at 24 and 48 hours post-treatment.

During treatment of the mice, clinical signs like reduction of spontaneous activity and apathy were observed.

The positive control (cyclophosphamide) induced the appropriate increase in micronuclei. In the treated groups, there was no significant increase in the frequency of micronuclei in the bone marrow cells of mice after any treatment time.

In an unscheduled DNA synthesis assay (RCC 1160600, 2008), performed according to OECD guideline 486, 4 male Wistar rats per dose were treated once by gavage with the test substance, dissolved in polyethylene glycol, at doses of 1000 and 2000 mg/kg bw. After the treatment period of 4 and 16 hours, respectively, primary hepatocyte cultures were established and evaluated.

During treatment of the rats, clinical signs like reduction of spontaneous activity and ruffled fur observed. Both effects were reversible 24 hours after administration.

The positive controls (N,N’-dimethylhydrazinedihydrochloride, 2-acetylaminofluorene) induced the appropriate response of unscheduled DNA synthesis. In the treated groups, there was no significant increase in the frequency of unscheduled DNA synthesis in primary hepatocytes of rats after any treatment time.

Short description of key information:
Genetic toxicity in vitro:
Ames test: negative (OECD Guideline Study 471, RCC 570401, 1996)
Chromosome aberration: ambiguous (OECD Guideline Study 473, RCC 570402, 1996)
Genetic toxicity in vivo:
Micronucleus: negative (OECD Guideline Study 474, RCC 582100, 1997)
UDS: negative (OECD Guideline Study 486, RCC 1160600, 1997)

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available studies are considered reliable and suitable for classification purposes under 67/548/EEC. As a result the substance is not considered to be classified for mutagenicity under Directive 67/548/EEC, as amended for the 28th time in Directive 2001/59/EC.

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

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for mutagenicity under Regulation (EC) No. 1272/2008, as amended for the second time in Directive (EC 286/2011).