Esterification products of Grape seeds, Vitis vinifera L. (Vitaceae), extract with hexadecanoyl chloride

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

1

Reference substance name:

Esterification products of Grape seeds, Vitis vinifera L. (Vitaceae), extract with hexadecanoyl chloride

EC Number:

941-319-3

Molecular formula:

Unknown (Substance of Unknown or Variable Composition)

IUPAC Name:

Esterification products of Grape seeds, Vitis vinifera L. (Vitaceae), extract with hexadecanoyl chloride

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Palmitoyl grape seed extract is the chemical name of BERKEMYOL PEPIN DE RAISIN (the trade name).

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

Commercially available post-mitochondrial fraction (S9) from livers of rodents treated with the enzyme inducing agent Aroclor

Test concentrations with justification for top dose:

Cytotoxicity evaluation of the test item was performed on 5 concentarions prepared by 1:3 serial dilutions starting at 5 (C5) up to 0.06 (C1) mg/plate, based on the solubility profile of the test item. Concentartions C4 to C1 ware prepared by 1:3 serial dilutions in the selected solvent (Corn oil) from the C5 concentartion.

Details on test system and experimental conditions:

All the bacterial strains used in the Ames test carry a mutant gene that prevents them from synthetizing an essential amino acid. These strains may carry additional mutations which increase their sensitivity to different types of mutagens. All S.typhimurium strains used in the test carry the rfa mutation. This mutation causes an alteration in the lipopolysaccharide (LPS) layer making the bacteria more permeable to larger molecules. The uvrB and uvrA deletions eliminate the accurate excision repair mechanism resulting in an increase in the rate of mutations due to an alternative DNA repair mechanism. The plasmid pKM101 in several strains enhances the chemical mutagenesis via an increase in the error-prone recombinational DNA repair mechanism.

The bacterial strains used for the study were grown from controlled Working Banks obtained from Master Banks (generated in Vivotecnia) in nutrient broth supplemented with the corresponding antibiotics when required.

Evaluation criteria:

The criteria used for determining a positive result involved taking into account a dose-response effect in the range tested and /or a reproducible increase at on or more concentrations in the number of revertant colonies per plate in at least one strain with or without metabolic activation system.

Results and discussion

Test results

Key result

Species / strain:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Applicant's summary and conclusion

Conclusions:

Based on the results obtained in this study (refer to attached study report), it can be concluded that the test item does not induce point mutations or frame-shifts in the genome of bacterial strains with or without metabolic activation regardless of the procedure.

Therefore, the test item BERKEMYOL PEPINS DE RAISIN at an exposure dose range of 5 - 0.06 mg/plate is considered to be NON MUTAGENIC / NON PRO-MUTAGENIC under the experimental conditions assayed.

Executive summary:

The bacterial reverse mutation test (Ames test) assesses the mutagenic or promutagenic ptential of the test item in the several bacterial strains.

The test was performed in accordance with OECD Guideline 471 for the Testing of Chemicals (Bacterial Reverse Mutation Test, adopted 21st July 1997) and the test Method B13/B14 of Commission Directive 2000/32/EC.

No cyototoxic activity was observed at a test item concentration of 5 mg/plate.

Five test item doses ranging from 5 and 0.06 mg/plate were assayed. None of the concentrations assayed for the test item showed an increase in the R value either with or without S9 metabolic activation regardless of the procedure.

No dose response for the test item was observed in any of the tested bacterial strains.

Based on the results obtained in this study, it can be concluded that the test item does note induce point mutations or frame-shifts in the genome of the bacterial strains with or without metabolic activation regardless of the procedure.

Therefore, the test item (BERKEMYOL PEPINS DE RAISIN) is not considered to be non mutagenic / non pro-mutagenic under the experimental conditions assayed.