Rape oil, reaction products with diethanolamine

Endpoint summary

Additional information

Reliable good quality and valid in vitro and in vivo studies covering the required endpoints of bacterial and mammalian mutagenicity as well as mammalian clastogenicity are available to assess the genotoxicity potential of the structural analogue amides, C18(unsatd.), N, N-bis(hydroxyethyl).

In vitro:

A study was conducted by NTP to determine the mutagenic potential of the read across substance oleic acid diethanolamine condensate (ODEA). Salmonella typhimurium strains TA97, TA98, TA100 and TA1535 were treated with the test substance using the Ames plate incorporation method at up to five dose levels for each bacterial strain, in triplicate, both with and without the addition of S9 mix. The dose range was 0.1 to 10 µg/plate (-S9 -mix) and 3.3 to 200 µg/plate (+S9 -mix). Cytotoxicity was observed at ≥3.3 µg/plate without metabolic activation and at 200 µg/plate with metabolic activation.No significant increase in the frequency of revertant colonies was recorded for any of the bacterial strains with any dose of the test substance, either with or without metabolic activation. The vehicle (ethanol) control or the negative control plates produced counts of revertant colonies within the normal range. All the positive control chemicals used in the test produced marked increases in the frequency of revertant colonies, both with and without the S9 -mix. Under the test conditions, the test substance was not mutagenic in Salmonella typhimurium strain TA97, TA98, TA100, or TA1535, with or without S9 metabolic activation.

 

Another study was performed by NTP to investigate the potential of the read across substance oleic acid diethanolamine condensate (ODEA) to induce mutations at the mouse lymphoma thymidine kinase locus using the cell line L5178Y. The assay was performed both with and without rat liver microsomal activation. Without metabolic activation, the test substance was treated at 0, 1.25, 2.5, 5 and 7.5 nL/mL in trial 1; 0, 2, 3, 4, 6, 8 and 12 nL/mL in trial 2 and 0, 3, 4, 6, 8, 12, 15 and 20 nL/mL in trial 3; with metabolic activation, at 0, 2.5, 5, 7.5, 10 and 15 nL/mL in trial 1 and at 0, 2.5, 5, 7.5, 10, 15 and 20 nL/mL in trial 2 for 4 h. After the 48-hour expression period, cells were plated in medium and soft agar supplemented with TFT for selection of TFT-resistant cells and cells were plated in nonselective medium and soft agar to determine cloning efficiency. Under the test conditions, no increase in the frequency of mutant colonies of L5178Y mouse lymphoma cells was noted after exposure to test substance, with or without S9 (NTP report 481, 1999).

In vivo:

 

A study was conducted to assess the clastogenic potential of the read across substance oleic acid diethanolamine condensate (ODEA). The test substance was dermally applied for 13 wk at 0, 50, 100, 200, 400 and 800 mg/kg bw. Peripheral blood samples were obtained from male and female mice, and smears were immediately prepared and fixed in absolute methanol.Under the conditions of the study, the test substance did not increase the frequencies of micronucleated normochromatic erythrocytes (NCEs) in peripheral blood of both male and female mice at the end of 13 wk (NTP, 2001).

Another study was conducted to assess the genotoxicity (chromosomal aberration) of the structurally similar LDEA using the mouse peripheral blood micronucleus test. Test substance was applied dermally for 14 wk with a frequency of 5 exposures/wk. Peripheral blood samples were obtained from male and female mice, and smears were immediately prepared and fixed in absolute methanol. The methanol-fixed slides were stained with acridine orange and coded. Slides were scanned to determine the frequency of micronuclei in 2,000 normochromatic erythrocytes (NCEs) in each of five animals per dose group. No increase in the frequency of micronucleated normochromatic erythrocytes was observed in the test at any dose level.Under the test conditions, the test material did not increase the frequency of micronuclei in peripheral blood cells of mice(NTP report 480, 1999).

The following information is taken into account for any hazard / risk assessment:

The structural analogue ODEA at 0.1 to 200 μg/plate was not mutagenic in Salmonella typhimurium strain TA97, TA98, TA100, or TA1535, with or without S9 metabolic activation. In addition, no increase in the frequency of mutant colonies of L5178Y mouse lymphoma cells was observed after exposure to ODEA at 1.25 to 20 nL/mL doses, with or without S9. Further, no increase in the frequency of micronucleated normochromatic erythrocytes was observed in peripheral blood samples from male and female mice treated dermally with ODEA at doses of 50-800 mg/kg bw/day for 13 weeks.

Based on the above overall weight of evidence, amides, C18(unsatd.), N,N-bis(hydroxyethyl) is considered to be non-genotoxic. On this basis, it is assumed that HE Rape Oil, reaction product with diethanolamine will be non-genotoxic also.

Justification for selection of genetic toxicity endpoint
No study was selected, since results from all the in vitro and in vivo studies conducted on structural analogues were negative.

Short description of key information:
The read across substance oleic acid diethanolamine condensate (ODEA) at 0.1 to 200 μg/plate was not mutagenic in Salmonella typhimurium strain TA97, TA98, TA100, or TA1535, with or without S9 metabolic activation. In addition, no increase in the frequency of mutant colonies of L5178Y mouse lymphoma cells was observed after exposure to ODEA at 1.25 to 20 nL/mL doses, with or without S9. Further, no increase in the frequency of micronucleated normochromatic erythrocytes was observed in peripheral blood samples from male and female mice treated dermally with ODEA at doses of 50-800 mg/kg bw/day for 13 weeks.
Based on the above overall weight of evidence, the structural analogue amides, C18(unsatd.), N,N-bis(hydroxyethyl) is considered to be non-genotoxic.
On this basis, it is assumed that HE Rape Oil, reaction product with diethanolamine will be non-genotoxic also.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

The structural analogue amides, C18 -unsatd., N, N-bis(hydroxyethyl) in the form of ODEA was consistently not mutagenic or clastogenic in short-term in vitro and in vivo genetic toxicity tests. On this basis, it is assumed that HE Rape Oil, reaction product with diethanolamine will be non-genotoxic also.

Therefore no classification is required for genotoxicity according to EC criteria (67/548/EEC) and according to CLP criteria (EC 1272/2008).