[No public or meaningful name is available]

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Not mutagenic or genotoxic.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2002

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Justification for type of information:

Judged to be scientifically reliable and appropriate for risk assessment by the Expert Panel of dermatologists and scientists at the Cosmetics Ingredient Review, Washington, DC., U.S.A.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Principles of method if other than guideline:

OECD guideline method by GLP

GLP compliance:

not specified

Type of assay:

in vitro mammalian cell gene mutation tests using the thymidine kinase gene

Target gene:

TK

Species / strain / cell type:

mouse lymphoma L5178Y cells

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Test concentrations with justification for top dose:

concentrations ranged from 6 to 46 µg/ml (with and without metabolic activation)

Vehicle / solvent:

not specified

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

3-methylcholanthrene

ethylmethanesulphonate

Details on test system and experimental conditions:

Three experimental traisl were included. Mouse lymphoma cells were tested with and without metabolic activation.

Key result

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Conclusions:

The test material was reported by an expert panel for the U.S. Cosmetic Ingredient Review to be nonmutagenic in the mouse lymphoma gene mutation assay (MLA).

Reference 2

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: experimental result in a reputable laboratory; precedes establishment of guidelines and GLP

Justification for type of information:

An analogue approach is used for the hazard assessment of several human health endpoints. The hypothesis for the analogue approach is that data can be read-across from data-rich substances, namely acetyl tributyl citrate (ATBC, CAS 77-90-7), acetyl triethyl citrate (CAS 77-89-4) and triethyl citrate (TEC, CAS 77-93-0) to acetyl trihexyl citrate (ATHC, CAS 24817-92-3). The analogue approach is based on common breakdown products via physical and biological processes, and similar functional groups, according to Annex XI, Section 1.5, of Regulation EC No. 1907/2006. Read-across to ATHC is indicated in order to avoid unnecessary in vivo testing according to Article 25 of Regulation EC No. 1907/2006.
The analogue approach to evaluating the safety of triethyl citrate is adopted here, reflecting the approach used by various expert panels and authoritative bodies in their safety assessment of TEC, included JECFA, EFSA, U.S. FDA, EPA and CIR. The use of analogues for hazard evaluation is justified (Scenarios 1 and 2 of the RAAF, 2015) because the substances have common breakdown products via physical and biological processes, which reflects the similar functional groups in their chemical structure. The proposed analogues have similar functional groups, including: a citric acid (tricarboxylic acid) backbone, three short-chain alkyl esters, and an acetyl group (except for TEC). Other than the acetyl group, there are no other functional groups which may introduce additional toxicities. The substances display similar classification based on similar toxicities.
The target substance is expected to have essentially the same effect in the toxicity test/endpoint as does the source substance. Dose descriptors obtained for derivation of a DNEL are adequate and appropriate, and do not underestimate the hazards of the registered substance.
This information is adequate to fulfill the data requirements of Annex IX, to be the basis for classification and labelling decisions, and for risk assessment.

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

standard Ames assay method with S9 fractions from additional species

GLP compliance:

no

Remarks:

precedes establishment of GLP

Type of assay:

bacterial reverse mutation assay

Target gene:

histidine

Species / strain / cell type:

S. typhimurium TA 1535

Species / strain / cell type:

S. typhimurium TA 1537

Species / strain / cell type:

S. typhimurium TA 1538

Species / strain / cell type:

Saccharomyces cerevisiae

Details on mammalian cell type (if applicable):

strain D4

Metabolic activation:

with and without

Metabolic activation system:

rat, mouse and monkey S-9 liver fraction

Test concentrations with justification for top dose:

0.4 to 1.7%

Vehicle / solvent:

not specified

Untreated negative controls:

yes

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Details on test system and experimental conditions:

Two types of assays were performed, i.e. plate tests and suspension tests. Based on cell toxicity studies, concentrations from 0.4 to 1.7% were used as the dose levels in the mutagenicity tests.

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not valid

Positive controls validity:

not specified

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not valid

Positive controls validity:

not specified

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not valid

Positive controls validity:

not specified

Key result

Species / strain:

Saccharomyces cerevisiae

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not valid

Positive controls validity:

not specified

Remarks on result:

other: negative

Remarks:

Not mutagenic

Conclusions:

The results are negative, non-mutagenic.

The analogue triethyl citrate was tested in a reverse bacterial mutatation assay using Salmonella typhimurium strains 1535, 1537 and 1538, and Saccharomyces cervisiae D4, with and without S-9 liver fractions (metabolic activation) from rat, mouse and monkey. All results were negative (non-mutagenic). These data are applicable to the target substance, and are adequate for risk assessment and classification and labelling.

Reference 3

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Evaluated as reliable by several authoritative bodies, such as JECFA, EFSA, CSTEE, U.S. Environmental Protection Agency, Cosmetic Ingredient Review Panel, etc.

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Target gene:

none. whole chromosomes

Species / strain / cell type:

lymphocytes: rat

Metabolic activation:

with and without

Metabolic activation system:

rat liver S9

Test concentrations with justification for top dose:

not specified

Vehicle / solvent:

not specified

Untreated negative controls:

yes

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Remarks:

Assume all appropriate controls included in this guideline study.

Details on test system and experimental conditions:

Cultures were initiated by inoculating approximately 0.3 ml of whole blood/5 ml of culture medium. Cultures were set up in duplicate at each dose level and incubated at 37°C. Forty-eight hours after setting up of whole blood cultures, the rat lymphocytes were treated with the test substance for four hours at concentrations of 0 (negative control), 4, 13.3, 40, 133 and 400 µg/ml both with and without S-9 metabolic activation. S-9 liver homogenate prepared from Aroclor 1254-teated Sprague-Dawley rats was purchased from an outside vendor. S-9 mix was prepared fresh and used on the day of preparation. DMSO was used to dissolve the test substance and also was used as the negative control group substance. The final concentration of DMSO in the culture medium was 1%. Cultures treated with 1000 µg/ml ethyl methanesulfonate and 4.2 µg/ml cyclophosphamide were used as positive controls for the non-activation and activation assays, respectively. The cultures were harvested 24 hours after termination of treatment. Mitotic indices were determined as the number of cells in metaphase among 1000 cells/replicate and expressed as percentages. Based upon the data on mitotic indices, cultures treated with the test substance at concentrations of 50, 133 and 400 µg/ml were selected for determining the frequencies of chromosomal aberrations. A total of 100 cells were scored in each replicate in the treated and negative control cultures and 50 cells were scored in the positive control cultures. Only those metaphases that contained 42 chromosomes were scored with the exception of severely damaged cells (cells having ≥10 aberrations/cell), in which case, accurate counts of the chromosomes were not possible. Gaps were not included in calculations of total cytogenetic aberrations. The following aberrations were included in the evaluation: chromatid breaks, chromatid exchanges, chromosome breaks, and chromosome exchanges.

Statistics:

yes, no information on methods

Key result

Species / strain:

lymphocytes: rat

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

The test substance showed no evidence of mutagenic activity in the presence and absence of an S-9 metabolic activation system.

Remarks on result:

other: negative

Remarks:

non-clastogenic

Conclusions:

The test was negative for induction of genotoxicity.

An analogue substance, acetyl tributyl citrate, showed no evidence of clastogenic activity in the presence and absence of an S-9 metabolic activation system in cultured rat lymphocytes. Data can be read-across between the analogues and target substance based on common breakdown products. This is adequate to fulfill the information requirements, to be the basis for classification and labelling decisions, and for risk assessment.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

No studies required

Mode of Action Analysis / Human Relevance Framework

Not applicable

Justification for classification or non-classification

There is no evidence of mutagenicity or clastogenicity in studies of analogues of the registered substance. The substance is not classified according to Regulation EC. No. 1272/2008.