Registration Dossier

Administrative data

Key value for chemical safety assessment

Additional information

Justification for grouping of substances and read-across

There are no studies available to assess the potential of the substance Fatty acids, tall-oil, triesters with trimethylolpropane (CAS 94581-09-6), to induce effects on genetic toxicity. In order to fulfil the standard information requirements set out in Annex VII-VIII, 8.4.1-3 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 put 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).

Overview of genetic toxicity

CAS

In vitro gene mutation in bacteria

In vitro cytogenicity in mammalian cells

Mammalian gene mutation

94581-09-6

Target substance

RA: Formerly CAS 85005-23-8

RA: CAS 403507-18-6

RA: Formerly CAS 85186-89-6

RA: CAS 403507-18-6

RA: CAS 85186-89-6

Formerly 85005-23-8

negative

--

--

403507-18-6

negative

negative

--

Formerly 85186-89-6

negative

--

negative

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, tall-oil triester with trimethylolpropane (CAS 94581-09-6). A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

In vitro gene mutation in bacteria

Formerly CAS 85005-23-8

In an Ames test, Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 as well as E. coli WP2 uvr A were treated with the test substance Fatty acids, C16-18 (even numbered) and C18-unsatd., branched and linear, di and triesters with trimethylolpropane (Formerly CAS 85005-23-8) diluted in Tween 80 according to OECD Guideline 471 (Verspeek-Rip, 1997). Test substance concentrations of 3, 10, 33, 100, 333, 1000, 3330 or 5000 µg/plate were tested in triplicate in two indipendent experiment, both with and without the addition of a rat liver homogenate metabolising system (S9-mix). Precipitation of the test substance was observed at and above 3330 µg/plate. The test material caused no cytotoxicity up to the highest, precipitating dose. No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test material, either with or without metabolic activation. The value of the controls for TA1537, in the presence of S9-mix, was just outside the historical control range but not considered to affect the validity of the study. Thus, Fatty acids, C16 -18 (even numbered) and C18 -unsatd., branched and linear, di and triesters with trimethylolpropane did not induce point mutations by base-pair changes or frame-shifts in the genome of the strains tested.

CAS 403507-18-6

The mutagenic potential of Fatty acids, C16-18 and C18-unsatd., branched and linear ester with trimethylolpropane (CAS 403507-18-6) was tested in a reverse mutation assay comparable to OECD Guideline 471 and under GLP conditions (Bowles, 2002). Salmonella typhimurium strains TA1535, TA1537, TA98, TA100 and TA102 were used. Tester strains were incubated with test material dissolved in acetone at concentrations of 50, 150, 500, 1500 and 5000 µg/plate with and without the addition of a metabolic activation system (phenobarbitale and beta-naphthoflavone induced rat liver S9 mix). Vehicle and appropriate positive controls were included into the study design. Positive control materials induced statistically significant increases in the frequency of revertant colonies indicating the satisfactory performance of the test and the activity of the metabolizing system. No increase in the frequency of revertant colonies compared to concurrent vehicle controls was observed in all strains treated with the test substance, neither in the presence nor in the absence of metabolic activation. No cytotoxicity was observed, but the test substance was tested up to precipitating concentrations. Thus, Fatty acids, C16-18 and C18-unsatd., branched and linear ester with trimethylolpropane did not induce point mutations by base-pair changes or frame-shifts in the genome of the strains tested.

Formerly CAS 85186-89-6

The mutagenic potential of Fatty acids, C8-18 and C18-unsatd., esters with trimethylolpropane (Formerly CAS 85186-89-6) was examined in a reverse mutation assay comparable to OECD Guideline 471 and under GLP conditions (Wiebel, 1999). Salmonella typhimurium strains TA1535, TA1537, TA98, TA100 and TA1538 were used. Tester strains were incubated with test material dissolved in acetone at concentrations of 8, 40, 200, 1000 and 5000 µg/plate (no toxicity but tested up to precipitating concentrations) with and without the addition of a metabolic activation system (phenobarbitale and beta-naphthoflavone induced rat liver S9 mix). Vehicle, negative and appropriate positive controls were included into the study design. Positive control materials induced statistically significant increases in the frequency of revertant colonies indicating the satisfactory performance of the test and the activity of the metabolizing system. No increase in the frequency of revertant colonies compared to concurrent controls was observed in all strains treated with the test substance, neither in the presence nor in the absence of metabolic activation. No cytotoxicity was observed but the test substance was tested up to limit concentrations. Thus, Fatty acids, C8-18 and C18-unsatd., esters with trimethylolpropane did not induce point mutations by base-pair changes or frame-shifts in the genome of the strains tested.

In vitro cytogenicity in mammalian cells

CAS 403507-18-6

An in vitro mammalian chromosome aberration test was performed with Fatty acids, C16-18 and C18-unsatd., branched and linear ester with trimethylolpropane (CAS 403507-18-6) in cultured peripheral human lymphocytes comparable to OECD Guideline 473 and under GLP conditions (Durward, 2004). Duplicate cultures of human lymphocytes were evaluated for chromosome aberrations in the presence and absence of metabolic activation (rat liver S9-mix). In the first experiment cells were exposed for 4 hours to the test substance dissolved in acetone at concentrations of 240, 320, 400 µg/mL with and without metabolic activation. In the second experiment cells were exposed for 4 hours to 240, 320, 400 µg/mL with metabolic activation and for 24 hours to 240, 320, 400 µg/mL followed by 24 hours expression time without metabolic activation. The test substance did not induce cytotoxicity but a cloudy precipitate was already visible at 40 µg/mL. Vehicle (solvent) controls induced aberration frequencies within the range expected for normal human lymphocytes. Mitomycin C and Cyclophosphamide were used as positive control materials inducing statistically significant increases in aberration frequencies indicating the satisfactory performance of the test and of the activity of the metabolizing system. Evaluation of 200 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.

In vitro gene mutation in mammalian cells

Formerly CAS 85186-89-6

An in vitro Mammalian Cell Gene Mutation Assay according to OECD Guideline 476 and GLP was performed with Fatty acids, C8-10(even), C14-18(even) and C16-18(even)-unsatd., triesters with trimethylolpropane (Formerly CAS 85186-89-6) in mouse lymphoma L5178Y cells (Verspeek-Rip, 2010). The cells were treated for 3 and 24 hours with 8% (v/v) and without S9-mix in the first experiment, respectively, and with 12% (v/v) with and without S9-mix in the second experiment, respectively. In the first experiment the test substance was tested at 0.3, 1, 3, 10, 33, 100, 333 and 750 μg/mL up to precipitation with 8% (v/v) and without S9-mix for 3 h. In the second experiment the test substance was tested at 0.3, 1, 3, 10, 33, 100, 333 and 750 μg/mL up to precipitation with 12% (v/v) for 3 hours and without S9-mix for 24 hours Cyclophosphamide and Methylmethanesulfonate were used as positive controls with and without S9 mix, respectively. No toxicity was observed and all dose levels were evaluated in the absence and presence of S9-mix. Positive and negative controls were valid and in 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 is not mutagenic in the mouse lymphoma L5178Y test system under the experimental conditions described.


Justification for selection of genetic toxicity endpoint
Hazard assessment is conducted by means of read-across from structural analogues. All available studies are adequate and reliable based on the identified similarities in structure and intrinsic properties between source and target substances and overall quality assessment (refer to the endpoint discussion for further details).

Short description of key information:
Negative results in Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537, TA 1538, TA 102 and E.coli WP uvr A with and without metabolic activation (OECD 471, GLP, analogue approach).
Negative results in mammalian citogenicity test using peripheral human lymphocytes (OECD 473, GLP, analogue approach)
Negative results in mammalian cell gene mutation test using mouse lymphoma L5178Y 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 the available data on the structural related substances 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.