Registration Dossier

Administrative data

Key value for chemical safety assessment

Additional information

Justification for grouping of substances and read-across

There are no data available for the in vitro mutagenicity in mammalian cells of Fatty acids, C14-18 and C16-18-unsatd., mixed esters with castor oil, castor oil fatty acids, 2-ethylhexanoic acid and 2,2-bis(hydroxymethyl)-1-butanol (CAS 92113-48-9). In order to fulfil the standard information requirements set out in Annex VIII, 8.4.3, , in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, read-across from a structurally related substance 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, 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, 1.5, of Regulation (EC) No 1907/2006, whereby physicochemical, toxicological and ecotoxicological properties may be predicted from data for reference substance(s) by interpolation to other substances on the basis of structural similarity, Fatty acids, C8-18 and C18-unsatd., esters with trimethylolpropane (CAS 85186-89-6) is selected as source substance for assessment of in vivo skin and eye irritation.

Genetic toxicity

CAS

92113-48-9 (a)

85186-89-6 (b)

Chemical name

Fatty acids, C14-18 and C16-18-unsatd., mixed esters with castor oil, castor oil fatty acids, 2-ethylhexanoic acid and 2,2-bis(hydroxymethyl)-1-butanol

Fatty acids, C8-18 and C18-unsatd., esters with trimethylolpropane

MW

512.8 – 975.5

512.76 -933.56 g/mol

Genetic Toxicity in vitro: gene mutation in bacteria

Experimental result:
negative

Experimental result:
negative

Genetic Toxicity in vitro: cytogenicity in mammalian cells

Experimental result:
negative

--

Genetic Toxicity in vitro: gene mutation in mammalian cells

RA: CAS 85186-89-6

Experimental result:
negative

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

(b) Reference (read-across) substances 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 the similarities in structure, properties and/or activities. The available endpoint information is used to predict the same endpoint for Fatty acids, C14-18 and C16-18-unsatd., mixed esters with castor oil, castor oil fatty acids, 2-ethylhexanoic acid and 2,2-bis(hydroxymethyl)-1-butanol (CAS 92113-48-9).

A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

Discussion

Gene mutation in bacteria

A study with Fatty acids, C14-18 and C16-18-unsatd., mixed esters with castor oil, castor oil fatty acids, 2-ethylhexanoic acid and 2,2-bis(hydroxymethyl)-1-butanol was performed in bacteria according to OECD guideline 471 and in compliance with GLP in the tester strains Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and E. coli WP2 uvrA (Verspeek-Rip, 2003). The test concentrations for the main study were in the range from 3 to 5000 µg/plate as determined in a preliminary toxicity study with and without metabolic activation in TA 100 and E. coli WP2 uvrA. The two independent experiments of the main study were performed each in triplicates with and without a metabolic activation system (Aroclor 1254-induced rat liver S9-mix). Cytotoxicity was determined by inspection of the bacterial background lawn and the increase in size of microcolonies. The positive and vehicle controls in the experiments showed the expected results and were therefore considered valid. No increase in the number of revertant colonies was noted in any of the bacterial strains, with and without metabolic activation system. From 1000 µg/plate upwards precipitation was observed with and without metabolic activation system, but no cytotoxicity was observed up to the highest, precipitating dose.

Under the conditions of this study Fatty acids, C14-18 and C16-18-unsatd., mixed esters with castor oil, castor oil fatty acids, 2-ethylhexanoic acid and 2,2-bis(hydroxymethyl)-1-butanol did not induce mutations in the bacterial mutation test in the absence and presence of a metabolic activation system in any of the strains tested.

In vitro mutagenicity in mammalian cells:

An in vitro Mammalian Cell Gene Mutation Assay according to OECD Guideline 476 and GLP was performed with Fatty acids, C8-18 and C18-unsatd., esters with trimethylolpropane (CAS 85186-89-6) in mouse lymphoma L5178Y cells (Verspeek-Rip, 2010). In the first experiment, the cells were treated for 3 hours with 0.3, 1, 3, 10, 33, 100, 333, and 750 µg/mL in the presence or absence of S9-mix (8% (v/v)). In the second experiment, test concentrations of 0.3, 1, 3, 10, 33, 100, 333, and 750 µg/mL were applied with metabolic activation (12%, v/v) for 3 h and 0.3, 1, 3, 10, 33, 100, 333, and 750 µg/mL without metabolic activation for 24 hours. The test substance was tested up to precipitating concentrations (333 µg/mL and above). Cyclophosphamide and methylmethanesulfonate were used as positive controls with and without S9-mix, respectively. No cytotoxicity was observed and all dose levels were evaluated in the absence and presence of S9-mix. Positive and negative controls were valid and in the range of historical control data. No significant increase in the mutation frequency at the TK locus was observed after treatment with the test substance either in the absence or in the presence of S9-mix. It was concluded that Fatty acids, C8-18 and C18-unsatd., esters with trimethylolpropane (CAS 85186-89-6) is not mutagenic in the mouse lymphoma L5178Y test system under the experimental conditions described.

In vitro cytogenicity in mammalian cells:

An in vitro mammalian chromosome aberration test was conducted with Fatty acids, C14-18 and C16-18-unsatd., mixed esters with castor oil, castor oil fatty acids, 2-ethylhexanoic acid and 2,2-bis(hydroxymethyl)-1-butanol in accordance with OECD guideline 473 under GLP conditions (Meerts, 2004). The induction of structural chromosome aberrations was evaluated in human lymphocytes in vitro incubated for 3 h with and without metabolic activation and 24 and 48 h without metabolic activation (S9-mix from rats treated with Aroclor 1245), respectively. Concentrations of 10, 33, 100 µg/mL (3 h incubation, with and without S9-mix) and 10, 33, 66, 100 µg/mL (24 and 48 h incubation, without S9-mix) of the test substance were applied. The vehicle used in the testing was DMSO. Cytotoxicity was evaluated by calculating the mitotic index. Precipitation was observed at 100 µg/mL and no cytotoxicity was observed. The vehicle and positive controls showed the expected results and were within the range of historical control data. No increase in the incidence of chromosome aberrations was observed under the conditions of the study. Thus, Fatty acids, C14-18 and C16-18-unsatd., mixed esters with castor oil, castor oil fatty acids, 2-ethylhexanoic acid and 2,2-bis(hydroxymethyl)-1-butanol did not show clastogenic activity in human lymphocytes in vitro under the conditions of the study.

Conclusion

Negative results in an Ames test and a chromosomal aberration test were observed with Fatty acids, C14-18 and C16-18-unsatd., mixed esters with castor oil, castor oil fatty acids, 2-ethylhexanoic acid and 2,2-bis(hydroxymethyl)-1-butanol . The analogue substance Fatty acids, C8-18 and C18-unsatd., esters with trimethylolpropane was negative in a mouse lymphoma assay. In conclusion, based on experimental data and read-across from an analogue substance Fatty acids, C14-18 and C16-18-unsatd., mixed esters with castor oil, castor oil fatty acids, 2-ethylhexanoic acid and 2,2-bis(hydroxymethyl)-1-butanol is considered to be not mutagenic and not clastogenic in vitro.


Justification for selection of genetic toxicity endpoint
Hazard assessment is conducted by means of substance specific data (OECD 471, OECD 473) and read-across from structural analogues (OECD 476). No study was selected, since all available in vitro genetic toxicity studies were negative. All available studies are adequate and reliable based on the identified similarities in structure and intrinsic properties between source and target substance and overall quality assessment (refer to the endpoint discussion for further details).

Short description of key information:
In vitro:
Mutagenicity in bacteria (OECD 471): negative
Cytogenicity in mammalian cells in a chromosome aberration test (OECD 473): negative
Mutagenicity in mammalian cells in a MLA assay (OECD 476): negative (based on read-across)

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Based on substance-specific data and read-across from the 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.