Registration Dossier

Diss Factsheets

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation : AMES TEST: The test substance, benzyl propionate did not induce mutagenicity in an Ames assay conducted on S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 strains with and without S9 activation.

From the comparison of the data obtained for the test and reference cigarettes, it was concluded that the addition of ingredients did not result in a positive mutagenic response in any of the strains under the conditions already described. Hence, the use of the tested ingredients had no influence on the mutagenic activity of the cigarettes.

Cytotoxicity: The test substance, benzyl propionate did not induce cytotoxicity in an neutral red uptake assay conducted on mouse embryo BALB/c 3T3 cells.

Link to relevant study records
Reference
Endpoint:
genetic toxicity in vitro
Remarks:
Type of genotoxicity: other: cytotoxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Qualifier:
according to guideline
Guideline:
other:
Principles of method if other than guideline:
The cytotoxicity of the gas/vapor phase and the particulate phase was determined in the neutral red uptake assay with mouse embryo BALB/c 3T3 cells.
GLP compliance:
no
Type of assay:
other: neutral red uptake assay
Species / strain / cell type:
mammalian cell line, other: mouse embryo BALB/c 3T3 cells
Additional strain / cell type characteristics:
not specified
Metabolic activation:
not specified
Test concentrations with justification for top dose:
eight concentrations up to approximately 160 µg TPM/ml medium
Vehicle / solvent:
DMSO
Untreated negative controls:
not specified
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:
Three TPM batches and three GVP batches were assayed for each cigarette type. For each batch, four replicate 96-well microtiter plates were used. Each concentration was replicated six times per microtiter plate. In each well, 10(4) cells were seeded and cultivated in culture medium containing 10% fetal bovine serum
(FBS). After 24 h the cells were exposed for 24 h to the smoke fractions dissolved in culture medium containing 4.8% FBS. At the end of the exposure period, the medium containing the smoke fractions was replaced with culture medium containing neutral red (25 mg/ml). Following a 3-h incubation period, 100 ml of an extraction solution (1% acetic acid in 50% ethanol) was added to each well and the absorbance of each well (directly correlated with the number of viable cells) was measured at 540 nm on a microtiter plate reader. To ensure the validity of the assay, TPM from the Reference Cigarette 1R4F and pure acrolein were both assayed in parallel.
Evaluation criteria:
The cytotoxic response was characterized as the EC50 value; that is, the effective concentration in mg TPM/ml culture medium that reduced the number of viable cells in the exposed culture by 50% compared to the untreated control. The measure of dosing, mg TPM/ml, refers either to the mass of the particulate phase itself or to the trapped gas/vapor phase constituents accompanying that particle mass.
Statistics:
Arithmetic means and measures of variance were calculated as descriptive statistics. The one-way analysis of variance was used to compare the results obtained for the control cigarette and those obtained for the test cigarettes containing the same group of ingredients. In those cases where this overall comparison showed a significant difference between the cigarettes, the Duncan test for pairwise comparison was applied. Results were considered to be statistically significant at P40.05 without adjustment for multiple testing
Species / strain:
mammalian cell line, other: mouse embryo BALB/c 3T3 cells
Metabolic activation:
not specified
Genotoxicity:
not specified
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:
This study demonstrates that within the sensitivity and the specificity of the test systems, the addition of the commonly used ingredients added did not increase the or cytotoxic activity of the resulting mainstream smoke, even at the exaggerated levels used.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Conclusions:
Interpretation of results (migrated information):
negative

The test substance, benzyl propionate did not induce cytotoxicity in an neutral red uptake assay conducted on mouse embryo BALB/c 3T3 cells.
Executive summary:

The test substance, benzyl propionate did not induce cytotoxicity in an neutral red uptake assay conducted on mouse embryo BALB/c 3T3 cells.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Gene mutation: AMES TEST: The test substance, benzyl propionate did not induce mutagenicity in an Ames assay conducted on S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 strains with and without S9 activation.

From the comparison of the data obtained for the test and reference cigarettes, it was concluded that the addition of ingredients did not result in a positive mutagenic response in any of the strains under the conditions already described. Hence, the use of the tested ingredients had no influence on the mutagenic activity of the cigarettes.

Cytotoxicity: The test substance, benzyl propionate did not induce cytotoxicity in an neutral red uptake assay conducted on mouse embryo BALB/c 3T3 cells.

DNA Damage: Benzyl propionate was not mutagenic to Bacillus subtilis M45 and B. subtilis H17 in the recombination assay at a concentration of 21 µg/disk.

Justification for selection of genetic toxicity endpoint

Data is from a reliable data source having Klimisch rating K2.

Justification for classification or non-classification

The substance, benzyl propionate, is not classified as genotoxic as it gives negative result in the cytotoxicity assay conducted on mouse embryo BALB/c3T3 cells as well as in the bacterial cell recombination assay.Although the Ames test gives positive result for some bacterial strains, it is due to the ingredients already in cigarette smoke and not due to the addition of the test substance. Hence, the substance is concluded to be non genotoxic in nature.

Categories Display