Fatty acids, tall-oil, esters with ethylene glycol

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

In vitro gene mutation in bacteria: Ames test (read-across, OECD 471): negative with and without metabolic activation in S. typhimurium TA1535, TA1537, TA98, TA100 and E. coli WP2 uvrA pKM

In vitro gene mutation in mammalian cells (read-across, OECD 476): negative in mouse lymphoma L5178Y cells with and without metabolic activation

The hazard assessment is based on the data currently available. New studies with the registered substance and/or other member substances of the glycol esters category will be conducted in the future. The finalised studies will be included in the technical dossier as soon as they become available and the hazard assessment will be re-evaluated accordingly.

For further details, please refer to the category concept document attached to the category object (linked under IUCLID section 0.2) showing an overview of the strategy for all substances within the glycol esters category.

Link to relevant study records

Reference

Reference 1

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 for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Key result

Species / strain:

S. typhimurium TA 1535

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:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1537

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 examined

True negative controls validity:

not examined

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 98

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 examined

True negative controls validity:

not examined

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium 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 examined

True negative controls validity:

not examined

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:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: Source: CAS 73138-39-3

The bacterial reverse mutation assay with the source substance Fatty acids, tall-oil, esters with ethylene glycol (CAS 73138 -39 -3) was selected as key study for reasons of data reliability and structural similarity.

Additional supporting data on mutagenicity in bacteria (Ames test) is given for the source substances Fatty acids, C18 and C18 unsatd., epoxidized, ester with ethylene glycol (CAS 151661-88-0), Fatty acids, C14-18 and C16-18-unsatd., esters with propylene glycol (CAS 84988-75-0) and Fatty acids, C16-18, esters with ethylene glycol (CAS 91031-31-1). The additional source substances were found not to be mutagenic in S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100, both in the presence and absence of metabolic activation.

Conclusions:

The read across approach is justified in the analogue justification. The target and source substance are considered unlikely to differ in their bacterial mutagenicity potential. Bacterial reverse mutation assays have been performed with the four analogue source substances in the presence and absence of metabolic activation. All results obtained were negative, i.e. no gene mutation in bacteria was observed. Therefore, no mutagenic potential in bacteria is expected for target substance Fatty acids, tall-oil, esters with ethylene glycol (68187-85-9).

Reference 2

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 for cross-reference:

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, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: Source: CAS 91031-31-1

Conclusions:

The read-across approach is justified in the analogue justification. The target and source substance are considered unlikely to differ in their genotoxic potential. In an in vitro mammalian cell gene mutation test using the thymidine kinase gene performed according to OECD guideline 476 with the source substance Fatty acids, C16-18, esters with ethylene glycol (CAS 91031-31-1) no genotoxicity was found in mouse lymphoma L5178Y cells. Therefore, no mutagenic potential in mammalian cells is expected for target substance Fatty acids, tall-oil, esters with ethylene glycol (68187-85-9).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

In vivo micronucleus assay (read-across, OECD 474): negative in mice

The hazard assessment is based on the data currently available. New studies with the registered substance and/or other member substances of the glycol esters category will be conducted in the future. The finalised studies will be included in the technical dossier as soon as they become available and the hazard assessment will be re-evaluated accordingly.

For further details, please refer to the category concept document attached to the category object (linked under IUCLID section 0.2) showing an overview of the strategy for all substances within the glycol esters category.

Link to relevant study records

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

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 for cross-reference:

read-across source

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: Source: CAS 151661-88-0

Conclusions:

The read-across approach is justified in the analogue justification. The target and source substance are considered unlikely to differ in their genotoxic potential. An in vivo mammalian erythrocyte micronucleus test (according to OECD guideline 474) was performed in male and female mice with the source substance Fatty acids, C18 and C18 unsatd., epoxidized, ester with ethylene glycol (CAS 151661-88-0). No increase in frequency of micronucleated polychromatic erythrocytes was found after a single oral gavage dose of 5000 mg/kg bw. Therefore, no in vivo genotoxic potential is expected for target substance Fatty acids, tall-oil, esters with ethylene glycol (68187-85-9).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Mode of Action Analysis / Human Relevance Framework

Not applicable.

Additional information

The hazard assessment is based on the data currently available. New studies with the registered substance and/or other member substances of the glycol esters category will be conducted in the future. The finalised studies will be included in the technical dossier as soon as they become available and the hazard assessment will be re-evaluated accordingly.

For further details, please refer to the category concept document attached to the category object (linked under IUCLID section 0.2) showing an overview of the strategy for all substances within the glycol esters category.

Genetic toxicity (mutagenicity) in bacteria in vitro

The mutagenic potential of Fatty acids, coco, esters with 1,3-butanediol (CAS 73138-39-3) was tested in a reverse mutation assay according to OECD guideline 471 under GLP conditions (key study, 2004). Salmonella typhimurium strains TA1535, TA1537, TA98, TA100 and E. coli WP2 uvrA pKM 101 were used in two experiments. Tester strains were incubated with test material at concentrations of 0, 50, 150, 500, 1500, and 5000 μg/plate with and without the addition of a metabolic activation system. No cytotoxicity or precipitation was reported. Vehicle and appropriate positive controls were included in the study design. Positive control materials induced 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 with concurrent vehicle and negative controls was observed in all strains treated with the test material, neither in the presence nor in the absence of metabolic activation. Fatty acids, coco, esters with 1,3-butanediol (CAS 73138-39-3) did not induce point mutations by base-pair changes or frame-shifts in the genome of the bacterial strains tested.

CAS 151661 -88-0

The in vitro mutagenicity of Fatty acids, C18 and C18 unsatd., epoxidized, ester with ethylene glycol (CAS 151661-88-0) was assessed in a bacterial reverse mutation study (Ames test), performed under GLP conditions according to OECD guideline 471 (supporting study, 1990). S. typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 1538 were exposed to the test substance at concentrations up to 5000 µg/plate in the first experiment and up to 2700 µg/plate in the second experiment. The vehicle, negative and positive controls were valid in the presence and absence of metabolic activation. In the first experiment without S9 mix, TA 1537 showed increased mutant colony rates up to 12-fold compared with the negative (solvent) control with the solvent acetone. This was judged to be a consequence of the observed cytotoxicity of acetone causing a reduced bacterial background lawn. In the second experiment with a narrower concentration range, no increase in colony rates was observed at all. It was concluded, that the results of TA 1537 in the first experiment were without biological significance. In summary it was concluded that the test substance did not induce reversions in the S. typhimurium strains, with or without metabolic activation.

CAS 91031-31-1

The in vitro mutagenicity of Fatty acids, C16-18, esters with ethylene glycol (CAS 91031-31-1) was assessed under GLP conditions in a bacterial reverse mutation study (Ames test) according to OECD guideline 471 (supporting study, 1991a) S. typhimurium strains TA 1535, TA 1537, TA98, TA100, and TA 1538 were exposed to the test substance at concentrations up to 5000 µg/plate in two experiments. At the highest concentration tested, precipitation occurred. No cytotoxicity was seen. The Vehicle, negative and positive controls were valid. In the presence and absence of metabolic activation the test substance did not induce reversions in the S. typhimurium strains.

CAS 84988-75-0

The in vitro genetic toxicity of Fatty acids, C14-18 and C16-18-unsatd., esters with propylene glycol (CAS 84988-75-0) was assessed in a bacterial reverse mutation study (Ames test), performed under GLP conditions according to OECD guideline 471 (supporting, 1991b). S. typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 1538 were exposed to the test substance at concentrations up to 5000 µg/plate. Precipitation was noted at the highest concentration, while no cytotoxicity was observed at any concentration level. The vehicle, negative and positive controls were valid in the presence and absence of metabolic activation. The test substance did not induce reversions in the S. typhimurium strains, with or without metabolic activation.

Genetic toxicity (mutagenicity) in mammalian cells in vitro

CAS 91031-31-1

The mutagenic properties of Fatty acids, C16-18, esters with ethylene glycol (CAS 91031-31-1) were assessed in an in vitro mammalian cell gene mutation study performed according to OECD guideline 476 under GLP conditions (key study, 2010). Gene mutations in the thymidine kinase locus were investigated in L5178Y mouse lymphoma cells in the presence and absence of a metabolic activation system (Phenobarbital/β-naphtoflavone-induced rat liver S9). In the first experiment, cells were exposed for 3 h to the test substance at concentrations of 0.1-333 µg/mL (in DMSO) with and without metabolic activation. In the second experiment, cells were exposed for 24 h to 3-175 µg/mL without metabolic activation, and for 3 h to 0.1-333 µg/mL with metabolic activation (12% S9-mix). The vehicle and positive controls in the study showed the expected results and were within the range of historical control data. Precipitation was noted at 100 µg/mL. Cytotoxicity (measured as relative total growth) was noted at 333 µg/mL during the 3-h treatment with and without metabolic activation, and at 100 µg/mL during the 24-h treatment without metabolic activation. There was no significant increase in the number of forward mutations at the thymidine kinase locus of L5178Y mouse lymphoma cells treated with the test material, neither in the presence nor in the absence of a metabolic activation system. Under the conditions of the study, Fatty acids, C16-18, esters with ethylene glycol did not show gene mutation activity in this test performed in L5178Y mouse lymphoma cells in vitro.

Genetic toxicity (micronucleus assay) in vivo

CAS 151661-88-0

A mammalian erythrocyte micronucleus test was performed under GLP conditions with Fatty acids, C18 and C18 unsatd., epoxidized, ester with ethylene glycol (CAS 151661-88-0) according to OECD guideline 474. Five mice/sex/group received a single oral gavage dose of 5000 mg/kg bw. Bone marrow was sampled from 5 male and 5 female mice at each of three samplings at 24, 48 and 72 hours after treatment. In each animal 1000 polychromatic erythrocytes were counted. No statistically significant increase in frequency of micro-nucleated polychromatic erythrocytes was found as compared with vehicle (arachis oil) and positive control (10 mg/kg bw cyclophosphamide) animals. No clinical signs were observed. Based on the study results, no in vivo genotoxic potential was found.

Overall conclusion for genetic toxicity

The analogue read-across from source substances was applied from in vitro studies on gene mutation in bacterial and mammalian cells and from an in vivo cytogenicity (micronucleus) study. The results of the available studies were consistently negative. Based on the available data from analogue substances, no mutagenic or clastogenic potential is expected for the target substance Fatty acids, tall-oil, esters with ethylene glycol (CAS 68187-85-9).

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". By applying the analogue concept to Fatty acids, tall-oil, esters with ethylene glycol (CAS 68187-85-9) data will be generated from analogue 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, including target substance data and 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.