Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames test (OECD 471): negative with and without metabolic activation in S. typhimurium TA 1535, TA 1537, TA 98 and TA 100, and E. coli WP2 uvrA pKM 101 (read-across)
Chromosome aberration (OECD 473): negative in primary human lymphocytes and CHO cells with and without metabolic activation (read-across)
Gene mutation in mammalian cells (OECD 476): negative in L5178Y mouse lymphoma cells with and without metabolic activation (read-across)

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Remarks:
Summary of available data used for the endpoint assessment of the target substance
Adequacy of study:
key study
Justification for type of information:
refer to analogue justification provided in IUCLID section 13
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Key result
Species / strain:
other: S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A pKM 101
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Remarks on result:
other: Source: CAS 95912-89-3

The available Ames test performed with the source substance CAS 95912-89-3 was selected as the key study for reasons of structural similarity to the registered substance and data reliability. Additional data with the source substance CAS 11138-60-6 were taken into account as supporting information. In this study, no significant increases in the frequency of revertant colonies were recorded for the tester strains S. typhimurium TA100, TA98, TA1535, TA1537 and TA1538 as well as for the tester strain E. coli WP2uvrA, with any dose of the test material, either with or without metabolic activation.

Conclusions:
The bacterial mutagenicity potential of the target substance is estimated based on an adequate and reliable in vitro study from the structural analogue source substances CAS 95912-89-3 and 11138-60-6, respectively. In the Ames studies, no mutagenic properties were determined in S. typhimurium strains TA1535, TA1537, TA98, TA100 and TA102, with and without metabolic activation. The read-across approach is justified in the analogue justification. The target and source substances are considered unlikely to differ in their mutagenic potential. The target substance Fatty acids, C8-10-(even numbered), esters with pentaerythritol and adipic acid is not expected to induce gene mutations in bacterial cells.
Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
refer to analogue justification provided in IUCLID section 13
Reason / purpose:
read-across source
Key result
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
other: Source: CAS 95912-89-3
Conclusions:
The mammalian mutagenicity potential of the target substance is estimated based on an adequate and reliable in vitro study from the structural analogue source substance CAS 95912-89-3. In the mouse lymphoma assay, the test substance did not induce an increase in the mutant frequency at any test substance concentration in the presence and absence of metabolic activation compared to controls. The read-across approach is justified in the analogue justification. The target and source substances are considered unlikely to differ in their mutagenic potential. The target substance Fatty acids, C8-10-(even numbered), esters with pentaerythritol and adipic acid is not expected to induce gene mutations in mammalian cells.
Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
refer to analogue justification provided in IUCLID section 13
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Key result
Species / strain:
lymphocytes: cultured peripheral human lymphocytes
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Key result
Species / strain:
Chinese hamster Ovary (CHO)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
48% in the highest dose for treatment of 4 h without metabolic activation, 33% with metabolic activation and 3% for 20 h exposure without metabolic activation
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: Source: CAS 95912-89-3
Conclusions:
The mammalian chromosome aberration potential of the target substance is estimated based on an adequate and reliable in vitro study from the structural analogue source substances CAS 95912-89-3 and 11138-60-6, respectively. In the mammalian chromosome aberration studies, no increase in the number of cells with chromosomal aberrations was observed compared to controls at any concentration in the presence and absence of metabolic activation. In addition, no polyploidy and no endoreduplication was noted in any of the treated cells. The read-across approach is justified in the analogue justification. The target and source substances are considered unlikely to differ in their mutagenic potential. The target substance Fatty acids, C8-10-(even numbered), esters with pentaerythritol and adipic acid is not expected a clastogen.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for read-across

Experimental data on in vitro mutagenicity and cytogenicity of Fatty acids, C8-10-(even numbered), esters with pentaerythritol and adipic acid are not available. The assessment was therefore based on studies conducted with analogue source 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 the analogue read-across approach is provided in the technical dossier (see IUCLID Section 13).

In vitro gene mutation in bacteria

CAS 95912-89-3

The potential mutagenicity of Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane (CAS 95912-89-3) was investigated in an Ames test (bacterial gene mutation assay) according to OECD 471 and in compliance with GLP (Key, 2010). The study was performed in two independent experiments in Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA 1537 and the Escherichia coli strain WP2 uvrA pKM 101 at concentrations of 0.06, 0.19, 0.56, 1.67 and 5 µL/plate, both in the absence and in the presence of liver microsomal activation system (S9 mix). In the first experiment, bacteria were incubated with the test substance concentrations for a period of 48 h using the plate incorporation method. In the second experiment, the same exposure duration was applied, but an additional preincubation period of 20 min was included in the assay. In both experiments, no cytotoxicity was observed up to the maximum concentration and the mean number of revertant colonies per plate was not increased at any test concentration. The positive controls included in both experiments showed the expected results.

Based on these results, Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane was not mutagenic in the presence and absence of metabolic activation in the selected strains of S. typhimurium (TA 98, TA 100, TA 1535 and TA 1537) and E. coli WP2 uvrA pKM 101 under the conditions of this bacterial mutation assay.

CAS 11138-60-6

In an Ames test conducted with the Fatty acids, 8-10 (even numbered), di- and triesters with propylidynetrimethanol (CAS 11138-60-6), Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98, TA 100 and E.coli WP2 uvr A were treated according to OECD 471 (Supporting, 1996). The test substance was diluted in ethanol and test substance concentrations of 0, 10, 33, 100, 333 and 1000 µg/plate were tested in triplicate, both with and without the addition of a rat liver homogenate metabolising system (S9). Precipitation of the test substance was observed at and above 100 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.

In vitro cytogenicity in mammalian cells

CAS 95912-89-3

The potential of Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane (CAS 95912-89-3) to induce chromosomal aberrations was investigated in a GLP-conform in vitro mammalian chromosome aberration test according to OECD 473 (WoE, 2012). Based on a preliminary cytotoxicity test, duplicate cultures of human peripheral lymphocytes were incubated with the test substance at concentrations of 312.5, 625, 1250, 2500 and 5000 µg/mL in two independent experiments. In the first experiment, cells were exposed to the test substance for a period of 4 h in the presence and absence of metabolic activation, while the duration of treatment in the second experiment was 4 h with S9 activation and 24 h without S9 activation, respectively. No signs of cytotoxicity were noted in both experiments each carried out without and with metabolic activation up to the maximum concentration of 5000 µg/mL. Based on these results, chromosome analyse was performed at concentrations of 625, 1250, 2500 and 5000 µg/mL. No increase in the number of cells with chromosomal aberrations was observed compared to controls at any concentration in the presence and absence of metabolic activation. In addition, no polyploidy and no endoreduplication was noted in any of the treated cells. The positive controls included in both experiments showed the expected results and thus verified the sensitivity of the assay.

Based on the negative results obtained in this chromosome aberration assay, it was concluded that Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane did not induce structural and numerical chromosomal aberrations in cultured peripheral human lymphocytes in the presence or absence of metabolic activation.

CAS 11138-60-6

An in vitro mammalian chromosome aberration test was performed with Fatty acids, 8-10 (even numbered), di- and triesters with propylidynetrimethanol (CAS 11138-60-6) in Chinese Hamster Ovary cells (CHO) according to OECD Guideline 473 and under GLP conditions (WoE, 1996). Duplicate cultures of CHO cells were evaluated for chromosome aberrations in the presence and absence of metabolic activation (Arochlor 1254 induced rat liver S9-mix). Cells were exposed for 4 and 20 hours without and for 4 hours with metabolic activation. The test substance was dissolved in ethanol and used at concentrations of 625, 1250, 2500, 5000 µg/mL. Cytotoxicity was observed at the highest dose tested regardless of metabolic activation. Mitomycin C and cyclophosphamide was used as positive control without and with metabolic activation respectively. Vehicle (solvent) controls induced aberration frequencies within the range expected. Positive control material induced 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 at any dose level tested in comparison to the negative controls. The test material was therefore considered to be non-clastogenic to CHO cells in vitro.

In vitro gene mutation in mammalian cells

CAS 95912-89-3

To assess the potential mutagenicity of Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane (CAS 95912-89-3) in mammalian cells, a GLP-conform Mouse Lymphoma Assay was performed according to OECD 476 (Key, 2012). Based on the results of a preliminary cytoxicity test, mutations at the TK locus of mouse-lymphoma L5178Y cells were investigated at test substance concentrations ranging from 312.5 to 5000 µg/mL in two independent experiments. In the first experiment, cells were treated for a period of 3 h both in the presence and absence of metabolic activation (S9 mix). In the second experiment, the same treatment duration was used in the presence of S9 mix, whereas exposure to the test substance was extended to 24 h in the absence of S9 mix. After an expression period of 48 h in the presence of 5-trifluorothymidine (TFT) selective medium, the test substance did not induce an increase in the mutant frequency at any test substance concentration in the presence and absence of metabolic activation compared to controls. No cytotoxicity and no alterations in the ratio of small to large mutant colonies were observed at any concentrations with and without S9 activation between treated and control cultures. The positive controls significantly increased mutant frequency, demonstrating the sensitivity of the test system and the efficacy of the metabolic activation system.

In conclusion, Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane did not increase the mutant frequency in mouse-lymphoma L5178Y cells in the absence and presence of metabolic activation.

Overall conclusion

There is no study available on the in vitro mutagenicity and cytogenicity with the target substance Fatty acids, C8-10-(even numbered), esters with pentaerythritol and adipic acid. Therefore, analogue read-across from source substances was applied from in vitro studies on genetic toxicity in bacteria and mammalian cells using 2 source substances. The results of the available in vitro studies on source substances were consistently negative. Based on the available data and following the analogue approach, Fatty acids, C8-10-(even numbered), esters with pentaerythritol and adipic acid is considered to be nonmutagenic in vitro.

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 Fatty acids, C8-10-(even numbered), esters with pentaerythritol and adipic acid 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) No. 1272/2008 and are therefore conclusive but not sufficient for classification.