Fatty acids, tall-oil, reaction products with formaldehyde and (Z)-N-9-octadecenyl-1,3-propanediamine

Endpoint summary

Additional information

Genetic toxicity in vitro

Bacterial reverse mutation assay:

Wagner (1995) performed an Ames test (OECD 471 and EU B.13/14) with S. typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and E. coli strains WP2 uvrA pKM 101 and E. coli WP2 with and without metabolic activation.

Following test concentrations were applied in triplicate:

Preliminary Toxicity Assay: 6.7, 10, 33, 67, 100, 333, 667, 1000, 3333, 5000 µg/plate

Mutagenicity Assay: 10, 33, 100, 333, 1000, 5000 µg/plate

Negative controls and positive controls were run in triplicate and were considered to be valid. The test system was exposed to the test article via the plate incorporation methodology originally described by Ames et al. (1975) and updated by Maron and Ames (1983). In the conformatory assay, the test system was exposed to the test article via the preincubation methodology described by Yahagi et al. (1977). According to the results of the study, the test substance is not mutagenic in the Ames test with and without metabolic activation.

 

Chromosome Aberration:

Verbaan (2013) performed an in vitro Chromosome Aberration test in cultured peripheral human lymphocytes, blood collected from healthy adult, non-smoking, male volunteers (OECD 473 and EU Method B.10). Two experiments were performed using different test concentrations with and without S9 activation: 3h with and without S9 mix (first cytogenicity assay); 3 (with S9 mix) and 24 and 48h (without S9 mix) (second cytogenicity assay); 24 hours without S9 mix (cytogenicity assay 2A). Both negative and positive controls were considered to be valid. Reaction product of oleid acid, N1-(9Z)-9-octadecen-1-yl-1,3-propanediamine and paraformaldehyde is not clastogenic in human lymphocytes under the experimental conditions.

In vitro mammalian cell gene mutation:

Verspeek-Rip (2013) performed an in vitro mammalian gene mutation assay in L5178Y TK+/- lymphoma cells according to OECD Guideline 476, EU Method B.17 and IWGT recommendations published in literature. Two experiments were performed using different concentrations with and without metabolic S9 activation. Plates were tested in duplicate. Plates were exposed for 3 hours with and without S9, second experiment: 24 hours without S9 and 3 hours with S9. The positive and negative (vehicle) controls were considered to be valid. It was concluded that the test substance did not induce mutation at the TK locus of L5178Y mouse lymphoma cells in vitro in the absence or presence of S9 metabolic activation under the experimental conditions.

Genetic toxicity in vivo

According to REACH Annex IX section 8.4, column 2, no further in vivo testing is required as no positive results were obtained in any of the three in vitro studies performed according to REACH Annexes VII and VIII section 8.4.

Justification for selection of genetic toxicity endpoint
One study can not be selected as there are 3 in vitro key studies available.

Short description of key information:
Genetic toxicity in vitro:
Bacterial reverse mutation assay: performed according to OECD Guideline 471 in S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvr A (Wagner, 1995). The test item was not mutagenic with and without metabolic activation.

In vitro mammalian chromosome aberration test: performed according to OECD Guideline 473 and EU Method B.10 in cultured peripheral human lymphocytes (Verbaan, 2013). The test item was not clastogenic.

In vitro mammalian cell gene mutation test: the test item was tested for mutation using L5178Y mouse lymphoma cells. The test was performed according to OECD Guideline 476 and E.U Method B.17 (Verspeek, 2013). The test item was not mutagenic under the experimental conditions.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Based on the available data and according to the criteria of the DSD and CLP regulation, reaction product of oleic acid N1 -(9Z)-9 -octadecen-1 -YL-1,3 -propanediamine and paraformaldehide should not be classified for mutageniticity.