Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames test (OECD 471, GLP, read across): not mutagenic in TA 98, TA 100, TA 1535, TA 1537 and WP2 uvrA with and without metabolic activation

Micronucleus test (OECD 487, GLP, read across): not clastogenic in human lymphocytes with and without metabolic activation

Mouse lymphoma assay (OECD 476, GLP, read across): not mutagenic in mouse lymphoma L5178 cells with and without metabolic activation

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

There are no data available regarding genetic toxicity of the target substance Nonanoic acid, esters with adipic acid and trimethylolpropane. Therefore, read-across from the appropriate structural analogue substance trimethylolpropane tripelargonate (CAS 126-57-8) is conducted in accordance with Regulation (EC) No 1907/2006, Annex XI, 1.5. in order to fulfil the standard information requirements defined in Regulation (EC) No 1907/2006, Annex VII-VIII, 8.4. Common functional groups and structural similarities combined with similar toxicokinetic properties of the source and target substances 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).

Mutagenicity in bacterial cells

CAS 126-57-8

The potential mutagenicity of trimethylolpropane tripelargonate (CAS 126-57-8) was investigated in an Ames test (bacterial gene mutation assay) according to OECD guideline 471 and in compliance with GLP (RTC, 2013). 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 at 313, 625, 1250, 2500 and 5000 µg/plate in the absence of metabolic activation system and at concentrations of 100, 200, 400, 800 and 1600 µg/plate in the presence of metabolic activation system. No toxicity was observed with any tester strain at any dose level, in the absence or presence of S9 metabolism. Precipitation of the test item, which did not interfere with the scoring, was observed at the end of the incubation period at the highest concentrations in the presence of S9 metabolism only. The sterility of the S9 mix and of the test item solutions was confirmed by the absence of colonies on additional agar plates spread separately with these solutions. Marked increases in revertant numbers were obtained in these tests following treatment with the positive control items, indicating that the assay system was functioning correctly. No relevant increase in the number of revertant colonies was observed in the plate incorporation or pre-incubation assay, at any dose level, with any tester strain, in the absence or presence of S9 metabolism.

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 guideline 471 and in compliance with GLP (Nyco, 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. The positive controls included in both experiments showed the expected results. 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.

Cytogenicity in mammalian cells in vitro

CAS 126-57-8

Trimethylolpropane tripelargonate (CAS 126-57-8) was assayed for the ability to induce micronuclei in human lymphocytes, following in vitro treatment in the presence and absence of S9 metabolic activation. The test was performed similar to OECD guideline 487 and GLP (RTC, 2014). Two main experiments were performed. In the first experiment, the cells were treated for 3 hours in the presence and absence of S9 metabolism, respectively. The harvest time of 32 hours corresponding to approximately 2 cell cycles was used. As negative results were obtained, a second experiment was performed in the absence of S9 metabolism using approximately the same harvest time. A continuous treatment until harvest at 31 hours was used. Solutions of the test item were prepared in acetone. For the main experiment, the maximum dose level for treatment was selected in agreement with the study protocol and on the basis of the solubility of the test item. Dose levels of 4.06, 7.11, 12.4, 21.8, 38.1, 66.6, 117, 204, 357 and 625 µg/mL were used for the first main experiment. Based on the results obtained, dose levels of 4.06, 7.11, 12.4, 21.8, 38.1, 66.6, 117 and 204 µg/mL were used for the second main experiment. Each experiment included appropriate negative and positive controls. Two replicate cell cultures were prepared at each test point. The actin polymerisation inhibitor cytochalasin B was added prior to the targeted mitosis to allow the selective analysis of micronucleus frequency in binucleated cells. Dose levels were selected for the scoring of micronuclei taking into account the cytotoxicity of the test item treatments, calculated by the cytokinesis-block proliferation index (CBPI), and the observed test item precipitation by the end of treatment. 1000 binucleated cells per culture were scored to assess the frequency of micronucleated cells. Statistically increase in the incidence of micronuclei cells were observed following treatments with the positive controls Cyclosphosphamide, Motomycin-C and Colchicine, showing the validity of the test system.Following treatment with the test item, no statistically significant increase in the incidence of micronucleated cells over the concurrent vehicle control value was observed at any dose level in any treatment series.

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 performed according to OECD guideline 473 (Oleon, 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. The positive controls included in both experiments showed the expected results and thus verified the sensitivity of the assay. 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. 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.

Mutagenicity in mammalian cells in vitro

CAS 126-57-8

The test item Trimethylolpropane tripelargonate (CAS 126-57-8) was examined for mutagenic activity in mouse lymphoma L5178 cells in the absence and presence of S9 metabolic activation. The test was performed According to OECD guideline 476 and GLP (RTC, 2014). Based on the results obtained in the preliminary toxicity trial, two independent assays for mutation to trifluorothymidine resistance were performed using the following dose levels: Assay n°1 with and without S9, treatment of 3 hours doses: 25, 79.1, 250, 791 and 2500, µg/mL; Assay n°2: without S9, treatment of 24 hours doses: 78.1, 156, 313, 625, 1250 and 2500 µg/mL; Assay n°3 with S9, treatment of 3 hours doses: 78.1, 156, 313, 625, 1250 and 2500 µg/mL. No relevant toxicity was observed at any concentration tested in any treatment series. In both experiments, at the end of the treatment time, test item particles in suspension were noted at the highest or two highest concentrations.. Negative and positive control treatments were included in each mutation experiment, both in the absence and presence of S9 metabolism. The mutant frequencies in the solvent control cultures fell within the normal range. Marked increases were obtained with the positive control treatments indicating the correct functioning of the assay system. No increases in mutant frequencies were observed following treatment with the test item, in the absence or presence of S9 metabolism. It was concluded that Trimethylolpropane tripelargonate (CAS 126-57-8) did not induce mutation at the TK locus of L5178Y mouse lymphoma cells in vitro in the absence or presence of S9 metabolic activation under the reported experimental conditions.

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 guideline 476 (Oleon, 2012). Based on the results of a preliminary cytotoxicity test, mutations at the TK locus of mouse-lymphoma L5178Y cells were investigated at test substance concentrations of 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.

Conclusion for genetic toxicity

In summary, in vitro studies investigating the genetic mutation in bacteria, cytogenicity in mammalian cells and mutagenicity in mammalian cells are available for both analogue substances, Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane (CAS 95912-89-3) and Trimethylolpropane tripelargonate (CAS 126-57-8), all providing negative results. Therefore, no genetic toxicity is expected for the target substance Nonanoic acid, esters with adipic acid and trimethylolpropane.

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 Nonanoic acid, esters with adipic acid and trimethylolpropane, 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 and mutagenicity do not meet the classification criteria according to Regulation (EC) 1272/2008 and are therefore conclusive but not sufficient for classification.