Fatty acids, C16-18, tetraesters with 3,3'-oxybis[1,2-propanediol]

Endpoint summary

Additional information

Justification for grouping of substances and read-across

There are no data available for the genetic toxicity of the substance Fatty acids, C16-18, tetraesters with 3,3'-oxybis[1,2-propanediol] (CAS# 91050-80-5). In order to fulfil the standard information requirements set out in Annex VII and VIII, 8.4, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, read-across from structurally related substances was conducted.

In accordance with Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met. In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests", which includes the use of information from structurally related substances (grouping or read-across).

Having regard to the general rules for grouping of substances and read-across approach laid down in Annex XI, Item 1.5, of Regulation (EC) No 1907/2006, whereby toxicological properties may be predicted from data for the reference substance(s) on the basis of structural similarity, the substances listed below are selected as reference substances for assessment of toxicological endpoints, for which information gaps are identified.

Overview of genetic toxicity

CAS	Bacterial gene mutation	Cytogenicity in mammalian cells in vitro	Mammalian gene mutation
91050-80-5(a)	RA: 49553-76-6	RA: 49553-76-6	RA: 49553-76-6
49553-76-6(b)	negative	negative	negative

(a) Substances subject to the REACh  Phase-in registration deadline of 31 May 2013 are indicated in bold font. Only for this substance a full set of experimental results and/or read-across is given.

 (b) Substances that are either already registered under REACh or not subject to the REACh  Phase-in registration deadline of 31 May 2013 are indicated in normal font. Lack of data for a given endpoint is indicated by “--“.

 

The above mentioned substances are considered to be similar on the basis of structural similarity resulting in similar properties and/or activities. The available endpoint information is used to predict the same endpoints for Fatty acids, C16-18, tetraesters with 3,3'-oxybis[1,2-propanediol] (CAS# 91050-80-5). A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

 

Discussion

In vitro gene mutation study in bacteria

Since no studies investigating the in vitro gene mutation in bacteria of Fatty acids, C16-18, tetraesters with 3,3'-oxybis[1,2-propanediol] (CAS# 91050-80-5) are available, in accordance with Regulation (EC) No. 1907/2006 Annex XI, 1.5, a read-across from the structurally related analogue substances oleic acid, monoester with oxybis(propanediol) (CAS# 49553-76-6), was conducted.

CAS 49553-76-6

A bacterial gene mutation assay (Ames test) was performed with oleic acid, monoester with oxybis(propanediol), (CAS# 49553-76-6) according to OECD Guideline 471 and under GLP conditions (Myhre, 2012). The plate incorporation procedure was performed with Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 and E. coli WP2uver A strain in the absence and presence of metabolic activation (Aroclor 1254-induced rat liver S9-mix). A preliminary cytotoxicity test and a main assay were conducted each in triplicates at concentrations from 33.3 to 5000 µg/plate and 333 to 5000 µg/plate respectively (vehicle: DMSO). No increase in the number of revertant colonies was noted in any of the bacterial strains, with and without metabolic activation system. No cytotoxicity was observed up to the highest dose tested. The included positive and negative controls showed the expected results. Under the study conditions, the test substance did not induce mutations in the bacterial mutation assay in the absence and presence of a metabolic activation system in any of the strains tested.

Genetic toxicity (mutagenicity) in mammalian cells in-vitro

Since no studies investigating the in vitro gene mutation in mammalian cells of Fatty acids, C16-18, tetraesters with 3,3'-oxybis[1,2-propanediol] (CAS# 91050-80-5) are available, in accordance with Regulation (EC) No. 1907/2006 Annex XI, 1.5, a read-across from the structurally related analogue substances oleic acid, monoester with oxybis(propanediol) (CAS# 49553-76-6), was conducted.

 

CAS# 49553-76-6

The in vitro mammalian cell gene mutation study of with oleic acid, monoester with oxybis(propanediol), (CAS# 49553-76-6) was carried out according to OECD Guideline 476 under GLP conditions (Clarke, 2012). Gene mutations in the thymidine kinase locus were investigated in L5178Y mouse lymphoma cells in the presence and absence of a metabolic activation system (Aroclor 1254 rat liver S9). In the preliminary toxicity assay, the maximum concentration of test substance in medium was 5000 µg/mL. In the first experiment, cells were exposed for 4 h to test substance at concentrations of 10-150 µg/mL (in DMSO) with and without metabolic activation. The concentrations chosen for cloning were 10, 20, 30, 40 and 50 µg/mL in the presence and absence of metabolic activation. No cloned cultures exhibited induced mutant frequencies ≥ 90 mutants per 10 E6 clonable cells.

Concentrations of the second experiment without metabolic activation for an exposure time of 24 h ranged from 5-75 µg/mL. The concentrations chosen for cloning were 5, 10, 20, 30 and 40 µg/mL. No cloned cultures exhibited induced mutant frequencies ≥ 90 mutants per 10E6 clonable cells. The vehicle and positive controls in the study showed the expected results and were within the range of historical control data. Cytotoxicity was observed at concentrations ≥ 30 µg/mL without metabolic activation and ≥ 50 µg/mL with metabolic activation. There was no significant increase in the number of forward mutations at the thymidine kinase locus of L5178Y mouse lymphoma cells treated with the test material, neither in the presence nor in the absence of a metabolic activation system. Under the conditions of the study, oleic acid, monoester with oxybis(propanediol) did not show gene mutation activity in this test performed in L5178Y mouse lymphoma cells in vitro.

 

Genetic toxicity (cytogenicity) in mammalian cells in-vitro

Since no studies investigating the in vitro cytogenicity in mammalian cells of Fatty acids, C16-18, tetraesters with 3,3'-oxybis[1,2-propanediol] (CAS# 91050-80-5) are available, in accordance with Regulation (EC) No. 1907/2006 Annex XI, 1.5, a read-across from the structurally related analogue substances oleic acid, monoester with oxybis(propanediol) (CAS# 49553-76-6), was conducted.

CAS 49553-76-6

An in vitro mammalian chromosome aberration test was performed Oleic acid, monoester with oxybis(propanediol), (CAS# 49553-76-6)in cultured peripheral human lymphocytes comparable to OECD Guideline 473 and under GLP conditions (Glover, 2012). Duplicate cultures of human lymphocytes were evaluated for chromosome aberrations in the presence and absence of metabolic activation (rat liver S9-mix). Cells were exposed for 4 hours to the test substance dissolved in DMSO at concentrations of 25, 50, 100, 150, 200µg/mL without metabolic activation, for 4 hours at concentration of 50, 100, 200, 300 and 400 µg/mL with metabolic activation and for 22 hours without metabolic activation at concentration of 10, 25, 50, 75 and 100 µg/mL. The test substance induced cytotoxicity at 150 μg/mL in the 4-hour non activated test condition, at 100 μg/mL in the 22-hour non-activated test and in the 4-hour S9-activated test at 200 µg/mL and above. Vehicle (solvent) controls induced aberration frequencies within the range expected for normal human lymphocytes. Mitomycin C and Cyclophosphamide were used as positive control substances inducing statistically significant increases in aberration frequencies indicating the satisfactory performance of the test and of the activity of the metabolizing system. Evaluation of 100 well-spread metaphase cells from each culture for structural chromosomal aberrations revealed no increase in the frequency of chromosome aberrations and polyploid cells at any dose level tested in comparison to the negative controls. The test material was therefore considered to be non-clastogenic to human lymphocytes in vitro.

 

Conclusions

In summary, one study investigating the genetic mutation in bacteria in-vitro is available with oleic acid, monoester with oxybis(propanediol) (CAS# 49553-76-6)  providing negative results.

Furthermore, no mutagenicity in mammalian cells in-vitro was observed oleic acid, monoester with oxybis(propanediol), (CAS# 49553-76-6).

Short description of key information:
Negative results in Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and E.coli with and without metabolic activation (OECD 471, GLP, analogue approach).
Negative results in mammalian cell gene mutation tests using mouse lymphoma cells, with and without metabolic activation (OECD 476, GLP, analogue approach).

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Based on read-across from structurally similar substances, the available data on genetic toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.