1,2,3-propanetriyl tris[12-(acetoxy)octadecanoate]

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Key study conducted according to recognised testing guideline and with GLP certification.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 December 2010 - 03 January 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

21 July 1997

Deviations:

no

GLP compliance:

no

Remarks:

The study was reported throughly and the lack of GLP certification is considered not to effect the reliability of the study result.

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

Storage conditions: Room temperature
CPTC ID No.: MID-4853.01

Target gene:

Histidine locus

Species / strain / cell type:

S. typhimurium TA 97

Species / strain / cell type:

S. typhimurium TA 98

Species / strain / cell type:

S. typhimurium TA 100

Species / strain / cell type:

S. typhimurium TA 102

Species / strain / cell type:

S. typhimurium TA 1535

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver microsomes (S9)

Test concentrations with justification for top dose:

Test sample: 5.0, 1.0, 0.5, 0.1, 0.05 mg/plate (with and without metabolic activation).

Vehicle / solvent:

- Solvent: 2-Propanol
- Justification: The solubility ofthe test sample was tested in different solvents at 50 mg/mL concentration and found to
be soluble in 2-Propanol and was the solvent used to dissolve this test sample in the study.

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

mitomycin C

other: 2-aminoanthracene (w/S9), ICR 191 Acridine, Daunomycin

Details on test system and experimental conditions:

The bacterial reverse mutation assay was used to evaluate the mutagenic potential of the test sample at 5 concentrations of the test sample: 5.0, 1.0, 0.5, 0.1 and 0.05 mg. Testing was done with the appropriate solvent control and positive controls were plated with overnight cultures of the test systems (TA 97a, TA 98, TA 100, TA102, TA 1535) on selective minimal agar in the presence and absence ofAroclor-induced rat liver S9. All dose levels of the test sample, solvent controls and positive controls were plated in triplicate. (Refer to attachment A: Protocol M1 0-4853 for detailed test procedure).

Rationale for test conditions:

The Bacterial Reverse Mutation Assay is widely used to evaluate the mutagenic properties of chemicals.The test is based on the work of Dr. Bruce Ames and his coworkers and is commonly referred to as the Ames Test. Their studies involved the development of select histidine auxotrophs of S. typhimurium that are normally growth arrested due to mutations in a gene needed to produce the essential amino acid Histidine. in the absence of an external histidine source, the cells cannot grow to form colonies unless a reversion of the mutation occurs which allows the production of histidine to be resumed. As might be expected, Spontaneous reversions occur with each of the strains. However, chemical agents can induce a mutagenic response so that the number of revertant colonies is substantially higher than the spontaneous background reversion level. The test involves the analysis of the number of revertant colonies that are obtained with each strain in the presence and absence of the test chemical. Since the mutagenic response of a formulation could vary with the concentration, test articles are routinely dosed over an appropriate concentration range. in this-protocol, a complete set of positive and negative controls is included with each assay, and is plated routinely with all of the tester strains. AroclorTM 1254 induced rat liver microsomes are included to mimic the in viva activity of the liver enzymes in activating some pro-mutagens to mutagenic status.

Evaluation criteria:

Negative (solvent) Control Counts
The colonies that grew on the Minimal Glucose Agar plates developed from single cells that had regained their ability to grow in the absence of added histidine. The genetic reversion, from histidine auxotrophy to prototrophy, that enabled those cells to grow in the absence of exogenous histidine might have arisen spontaneously or as the result of a mutation induced by the treatments (see Maron & Ames, p. 181). It is important to realize that some of the colonies that arose in the positive control plates would have grown in the absence of treatment; they arose spontaneously. Accordingly, the negative (solvent) control colony counts constitute an important baseline in your evaluation of the test results. Unfortunately, the spontaneous reversion frequencies for the various tester strains can be quite variable - nevertheless, large deviations form the "normal" range of spontaneous reversion values may signal systematic problems with the assay.

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

The results in Tables 1 and 2 show that the test strains were sensitive to the positive control mutagens and had a spontaneous reversion rate well within the accepted values of each strain, indicating that under the test conditions, the strains were sensitive to the detection of potentially genotoxic agents. Test sample M10-4853.01 was not cytotoxic to the test system. The metabolic activation using the S9 activation mixture shows an active microsomal preparation.
Using the same test conditions, there was no detectable genotoxic activity associated with the five concentrations (5.0, 1.0, 0.5, 0.1, 0.05 mg) of test sample M10-4853.01 (Hetester HCA Lot #: G30074 Tables 1 and 2) either in the presence or absence of S9 enzyme activation.

Conclusions:

The test material failed to induce reverse mutation in any of the test concentrations and cell strains (Salmonella typhimurium) tested with and without metabolic activation.

Executive summary:

In this guideline (OECD 471) study, conducted without GLP certification, the test material did not induce mutagenic effects in any of the test concentrations or cell strains used, with and without metabolic activation. The test was conducted at 0.05, 0.1, 0.5, 1.0, and 5.0 mg/plate. Salmonella typhimurium was chosen as the model using strains 97, 98, 100, 102, 1535 with and without metabolic activation (Aroclor induced rat liver S9 mix).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

The registered substance failed to induce statistically significant reverse mutations in the AMES test. The registered substance does not meet the mutagen criteria of the Classification, Labelling, and Packaging (CLP) regulation (1272/2008).