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Administrative data

Key value for chemical safety assessment

Additional information

Justification for grouping of substances and read-across

There are no sufficient data available for the genotoxic properties of DiPE triisononanoate triethylhexanoate (CAS 68443-84-5). In order to fulfil the standard information requirements set out in Annex 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, 1.5, of Regulation (EC) No 1907/2006, whereby substances may be predicted as similar provided that their physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity.

Genetic toxicity

CAS

68443-84-5

647028-25-9

68424-31-7

15834-04-5

Chemical name

DiPE triisononanoate triethylhexanoate

Dipentaerythritol ester of nC5/iC9 acids

Fatty acids, C5-10, esters with pentaerythritol

2,2-bis[[(1-oxopentyl)oxy]methyl] propane-1,3-diyl divalerate

MW

1053.6 g/mol

983-1096 g/mol

612.9 g/mol

472.6 g/mol

Genetic Toxicity in vitro: gene mutation in bacteria

RA: CAS 647028-25-9

Experimental result: negative

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Genetic Toxicity in vitro: cytogenicity in mammalian cells

RA: CAS 647028-25-9

Experimental result: negative

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Genetic Toxicity in vitro: gene mutation in mammalian cells

RA: CAS 15834-04-5

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Experimental result: negative

Genetic Toxicity in vivo

RA: CAS 68424-31-7

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Experimental result: negative

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The above mentioned substances are considered to be similar on the basis of the structural similar properties and/or activities. The available endpoint information is used to predict the same endpoints forDiPE triisononanoate triethylhexanoate(CAS 68443-84-5).

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

Discussion

Genetic toxicity in vitro:

In vitro mutagenicity in bacteria

Mutagenic activity in bacteria can be assessed by use of data from the analogue substanceDipentaerythritol ester of nC5/iC9 acids, which was tested in a bacterial gene mutation assay (Ames test) performed according to OECD guideline 471 and in compliance with GLP (Thompson, 1999). The tester strains were Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and E. coli WP2 uvrA. The test concentrations for the main study were determined in a preliminary toxicity study with and without metabolic activation at concentrations from 0.15 up to 5000 µg/plate in TA 100 and E. coli WP2 uvrA. The first and second experiment of the main study were performed each in triplicates according to the plate incorporation procedure at concentrations up to 5000 µg/plate (vehicle: acetone) with and without a metabolic activation system (Aroclor 1254-induced rat liver S9-mix). Cytotoxicity was determined by inspection of the bacterial background lawn. The positive and negative 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. At 5000 µg/plate an oily precipitate was observed in all tested strains, but no cytotoxicity was observed up to the highest, precipitating dose.

Under the conditions of this study, the read-across substance 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 2,2-bis[[(1-oxopentyl)oxy]methyl]propane-1,3-diyl divalerate (CAS 15834-04-5) in mouse lymphoma L5178Y cells (Croda, 2010). In the first experiment, the cells were treated for 3 hours with 0.03, 0.1, 0.3, 1, 3, 10, 33, 100 µg/mL in the presence or absence of S9-mix (8% (v/v)). In the second experiment, test concentrations of 0.03, 0.1, 0.3, 1, 3, 10, 33, 100 µg/mL were applied with metabolic activation (12%, v/v) for 3 h and 0.1, 1, 3, 10, 33, 100, 200, 250 µg/mL without metabolic activation for 24 hours. The test substance was tested up to precipitating concentrations (100 µg/mL and above). 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 2,2-bis[[(1-oxopentyl)oxy]methyl]propane-1,3-diyl divalerate 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 the analogue substanceDipentaerythritol ester of nC5/iC9 acidsin accordance with OECD guideline 473 under GLP conditions (Wright, 2000). The induction of structural chromosome aberrations was evaluated in human lymphocytes in vitro incubated for 4 h with and without metabolic activation and 20 h without metabolic activation (S9-mix from rats treated with Aroclor 1245), respectively. Concentrations of 39.06 - 5000 µg/mL (4 h incubation, with and without S9-mix) and 156.25 - 5000 µg/mL (20 h incubation, without S9-mix) of the test substance were applied. The vehicle used in the testing was acetone. Cytotoxicity was evaluated by calculating the mitotic index of 2000 cells and cells were checked for polyploidy. There was a cloudy appearance of the test material at all concentrations in both treatment groups after 4 h exposure. The negative and positive controls showed the expected results and were within the range of historical control data. No cytotoxicity was observed up to the highest tested concentration. No increase in the incidence of chromosome aberrations was observed under the conditions of the study. Thus, the read-across substance did not show clastogenic activity in human lymphocytes in vitro under the conditions of the study.

Genetic toxicity in vivo

In vivo gene mutation:

An in vivo micronucleus assay of the structural analogue the Fatty acids, C5-10, esters with pentaerythritol (CAS 68424-31-7) in mice was carried out according to OECD guideline 474 under GLP conditions (Griffiths and Mackay, 1992). On the basis of a dose range finding study, the highest concentration for the main study was chosen to be 5000 mg/kg bw. A single intraperitoneal injection with this dosage in corn oil was given to groups of 5 male and 5 female mice. Bone marrow samples were taken 24 and 48 h after dosing. A concurrent negative control with the vehicle alone and a positive control group given cyclophosphamide (65 mg/kg bw) was included in the study. The negative and positive controls showed the expected results. The test material did not induce a statistically significant increase in the number of micronucleated polychromatic erythrocytes in the bone marrow of the animals. Comparison of the percentage of polychromatic erythrocytes showed no significant differences between the control and test group females. A small but significant decrease was, however, noted in male mice treated with the test material (24 h sampling). This small decrease was considered not to be biologically significant compared to the concurrent control values. Under the conditions of the study Fatty acids, C5-10, esters with pentaerythritol did not induce chromosomal mutations in the bone marrow of mice.

In conclusion, assessment of the available data together with the lack offunctional groups associated with genotoxicityindicates that there is no mutagenic potential ofTriisononanoic acid, triester with 2,2'-[oxybis(methylene)]bis[2-(hydroxymethyl)propane-1,3-diol] tris(2-ethylhexanoate).

 


Justification for selection of genetic toxicity endpoint
Hazard assessment is conducted by means of read-across from a structural analogue. 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:
Genetic toxicity in vitro:
RA-S, 647028-25-9, gene mutation (Ames test): S. typhimurium TA 1535, TA 1537, TA 98, and E. coli WP2 uvrA: negative with and without metabolic activation (according to OECD TG 471 and GLP)
RA-S, 647028-25-9, mammalian cytogenicity (chromosome aberration): human lymphocytes: negative (according to OECD TG 473 and GLP)
RA-S, 15834-04-5, mammalian mutagenicity (MLA): negative (according to OECD TG 476 and GLP)

Genetic toxicity in vivo:
RA-S, 647028-25-9, micronucleus test (bone marrow from mice): negative with and without metabolic activation (according to OECD TG 474 and GLP)

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

The available data on genetic toxicity of the test substance do not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.