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Key value for chemical safety assessment

Additional information

Justification for read-across

Data on the in vitro genetic toxicity in bacterial and mammalian cells of 2,2-bis(hydroxymethyl)propane-1,3-diyl didocosanoate (CAS 68258-72-0) are not available. The genetic toxicity assessment was therefore based on studies conducted with analogue substances as part of a read across approach, which is in accordance with Regulation (EC) No. 1907/2006, Annex XI, 1.5. For each specific endpoint the source substance(s) structurally closest to the target substance is/are chosen for read-across, with due regard to the requirements of adequacy and reliability of the available data. Structural similarities and similarities in properties and/or activities of the source and target substance are the basis of read-across. A detailed justification for analogue read-across approach is provided in the technical dossier (see IUCLID Section 13).

Genetic toxicity (mutagenicity) in bacteria in vitro

CAS 403507-18-6

The in vitro genetic toxicity of fatty acids, C16-18 and C18uns., branched and linear ester with trimethylolpropane was assessed in a bacterial reverse mutation assay (Ames test) performed according to OECD Guideline 471 and under GLP conditions (Bowles, 2002). The plate incorporation method was applied, using S. typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102. All the strains were tested at concentrations up to the limit value of 5000 µg /plate, with and without metabolic activation. No cytotoxicity was observed, while precipitation was observed at 5000 µg/plate. The controls were shown to be valid. The test substance did not induce an increase in reversions in the S. typhimurium strains, with or without metabolic activation and is considered to be not mutagenic under the conditions of this study.

CAS 85186-89-6

A bacterial reverse mutation assay (Ames test) was performed according to a protocol similar to OECD guideline 471 and under GLP conditions, using fatty acids, C8-18 and C18-unsatd., esters with trimethylolpropane (Wiebel, 1999).The plate incorporation method was applied, exposing S. typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100, to concentrations of 8 – 5000 µg /plate, with and without metabolic activation. No cytotoxicity or precipitation was observed. The positive and negative controls included for each tester strain were shown to be valid. No increase in the mean number of revertants per plate was observed when compared to controls. The test substance was considered to be not mutagenic under the conditions of this study.

CAS 85116-93-4

The potential mutagenicity of fatty acid C16-18, ester with pentaerythriol was assessed in five S. typhimurium strains (TA 1535, TA 1537, TA1538, TA 98 and TA 100) in an Ames test similar to OECD guideline 471 and under GLP conditions (Banduhn, 1991). Concentrations ranging from 8 to 5000 µg/plate were selected for treatment in the main assay, with and without metabolic activation. Precipitation was observed at concentrations from 1000 µg/plate. No increase in the mean number of revertants per plate was observed when compared to controls. The positive and negative controls included for each tester strain were shown to be valid. Based on the study results, the test substance was considered non-mutagenic in the selected strains of S. typhimurium in the presence and absence of metabolic activation.

 

Genetic toxicity (cytogenicity) in mammalian cells in vitro

CAS 403507-18-6

The cytogenetic potential of fatty acids, C16-18 and C18uns., branched and linear ester with trimethylolpropane was assessed in an in vitro mammalian chromosome aberration test in primary human lymphocytes, performed according to OECD guideline 473 and under GLP conditions (Durward, 2004). In the first experiment, cells were incubated with test substance concentrations of 40 - 400 µg/mL for 4 hours with a 24-hour fixation time, in the presence and absence of a metabolic activation system. In the second experiment cells were incubated with 40 - 400 µg/mL for 24 hours with a 24-hour fixation time, without metabolic activation. In the presence of metabolic activation cell were exposed to 40 - 400 µg/mL for 4 hours followed by a 24-hour fixation time. No cytotoxicity was observed. Metaphase analysis was performed on cells exposed to 240 – 400 µg/mL for both experiments. Precipitation was observed in the culture medium at 40 µg/mL in the 24-hour exposure trial and at 80 µg/mL in the 4-hour exposure trials. The vehicle (solvent) and positive controls were shown to be valid. The test material did not induce a statistically significant increase in the frequency of cells with chromosome aberrations, with or without metabolic activation.

Genetic toxicity (mutagenicity) in mammalian cells in vitro

CAS 85186-89-6

An in vitro mammalian cell gene mutation assay was performed using fatty acids, C8-18 and C18-unsatd., esters with trimethylolpropane, according to OECD guideline 476 and under GLP conditions (Verspeek-Rip, 2010). Mouse lymphoma L5178Y cells were treated with 0.3 - 750 µg/mL for 3 hours with and without metabolic activation in the first experiment. In the second experiment, cells were treated with 0.3 - 750 µg/mL for 3 hours with and for 24 hours without metabolic activation. After an expression time of 48 hours in growth medium, cells were incubated for 11-12 days. Precipitation was seen at concentrations of 333 µg/mL and above. The positive and negative controls were valid and within the range of historical control data. No cytotoxicity was observed. No significant increase in mutation frequency was observed, with and without metabolic activation.

 

Overall conclusion for genetic toxicity

There are no available studies on the in vitro genetic toxicity of the target substance 2,2-bis(hydroxymethyl)propane-1,3-diyl didocosanoate. Therefore analogue read-across from source substances was applied from in vitro studies on bacterial and mammalian cells, using 3 source substances. The results of the available in vitro studies on source substances were negative. Based on the available data on target and source substances, and following the analogue approach, 2,2-bis(hydroxymethyl)propane-1,3-diyl didocosanoate is not expected to be mutagenic and clastogenic in vitro.


Justification for selection of genetic toxicity endpoint
Hazard assessment is conducted by means of read-across from structural analogues. 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 the source and target substances and overall quality assessment (refer to the endpoint discussion for further details).

Short description of key information:
Genetic toxicity in vitro:
Ames test (OECD 471): negative with and without metabolic activation in S. typhimurium strains TA 1535, TA 1537, TA 1538, TA 98, TA 100 and TA 102
Chromosome aberration (OECD 473): negative in primary human lymphocytes cells with and without metabolic activation
Gene mutation in mammalian cells (OECD 476): negative in mouse lymphoma L5178Y cells with and without metabolic activation

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

According to Article 13 of Regulation (EC) No. 1907/2006 "General Requirements for Generation of Information on Intrinsic Properties of substances", information on intrinsic properties of substances may be generated by means other than tests e.g. from information from structurally related substances (grouping or read-across), provided that conditions set out in Annex XI are met. Annex XI, "General rules for adaptation of this standard testing regime set out in Annexes VII to X” states that “substances whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity may be considered as a group, or ‘category’ of substances. This avoids the need to test every substance for every endpoint". Since the analogue concept is applied to 2,2-bis(hydroxymethyl)propane-1,3-diyl didocosanoate (CAS 68258-72-0), data will be generated from data for reference source substance(s) to avoid unnecessary animal testing. Additionally, once the analogue read-across concept is applied, substances will be classified and labelled on this basis.

Therefore, based on the analogue read-across approach, the available data on genetic toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008, and are therefore conclusive but not sufficient for classification.