Anisyl formate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation toxicity study was performed by E. Roemera et al. ( Food and Chemical Toxicology 2002) to determine the mutagenic nature of Anisyl Formate IUPAC : 4-methoxyphenyl)methyl formate (122-91-8) using Salmonella typhimurium strains. Genetic toxicity in vitro study was assessed for Anisyl Formate(122-91-8) to evaluate its possible mutagenic potential .For this purpose AMES assay was performed according to guideline OECD 471.Anisyl Formate was exposed toS. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 strains with and without rat liver S9 activation byplate incorporation assay at the concentration of0, 1, 2, 3, 4, 5 and 5 mg/plate. DMSO was used as solvent. Negative and positive control were used .Anisyl Formate 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 rat liver S9 activation byplate incorporation assay .Although positive response is observed in some strains, there were also no apparent trends discernible that would suggest a change in mutagenic activity caused by the addition of the ingredient chemicals. Therefore Anisyl Formate(122-91-8) was not likely to classify as gene mutant in vitro.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from publication.

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

In vitro genotoxicity study of cigarette smoke (Anisyl Formate) by using Salmonella plate incorporation (Ames) assay.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

Details on test material
- Name of test material (as cited in study report): Anisyl Formate
- Molecular formula: C9H10O3
- Molecular weight: 166.175 g/mole
- Substance type: Organic
- Physical state: Liquid

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium, other: TA98, TA100, TA102, TA1535 and TA1537.

Details on mammalian cell type (if applicable):

Not applicable.

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

Not applicable.

Metabolic activation:

with and without

Metabolic activation system:

Metabolic activation system consisting of the postmitochondrial fraction of the livers from rats treated with Aroclor 1254

Test concentrations with justification for top dose:

0, 1, 2, 3, 4, 5 and 5 mg/plate

Vehicle / solvent:

DMSO

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

Details on test system and experimental conditions:

Details on test system and conditions
METHOD OF APPLICATION: In agar (plate incorporation)

DURATION
- Preincubation period: No data available
- Exposure duration: 44–48 h.
- Expression time (cells in growth medium): No data available
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): Mouse embryo BALB/c 3T3 cells

NUMBER OF REPLICATIONS: 3

NUMBER OF CELLS EVALUATED: 5

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: Neutral red uptake assay

OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available

OTHER: No data available

Rationale for test conditions:

Not specified

Evaluation criteria:

Number of His+ revertant colonies was determined

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 (Duncan, 1955) for pairwise comparison was applied. Results were considered to be statistically significant at P40.05 without adjustment for multiple testing.

Key result

Species / strain:

S. typhimurium, other: TA98, TA100, TA102, TA1535 and TA1537.

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:

COMPARISON WITH HISTORICAL CONTROL DATA: Yes

Conclusions:

Anisyl Formate 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 rat liver S9 activation.

Executive summary:

Genetic toxicity in vitro study was assessed forAnisyl Formate(122-91-8) to evaluate its possible mutagenic potential .For this purpose AMES assay was performed according to guideline OECD 471.Anisyl Formate was exposed toS. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 strains with and without rat liver S9 activation byplate incorporation assay at the concentration of0, 1, 2, 3, 4, 5 and 5 mg/plate. DMSO was used as solvent. Negative and positive control were used .Anisyl Formatedid 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 rat liver S9 activation byplate incorporation assay .Although positive response is observed in some strains, there were also no apparent trends discernible that would suggest a change in mutagenic activity caused by the addition of the ingredient chemicals. ThereforeAnisyl Formate(122-91-8) was not likely to classify as gene mutant in vitro.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Genotoxicity In-vitro

Various publications were reviewed to determine the mutagenic nature of 4-methoxyphenyl)methyl formate (122-91-8). The studies are as mentioned below:

Gene mutation toxicity study was performed by E. Roemera et al. ( Food and Chemical Toxicology 2002) to determine the mutagenic nature of Anisyl Formate IUPAC : 4-methoxyphenyl)methyl formate (122-91-8) using Salmonella typhimurium strains. Genetic toxicity in vitro study was assessed for Anisyl Formate(122-91-8) to evaluate its possible mutagenic potential .For this purpose AMES assay was performed according to guideline OECD 471.Anisyl Formate was exposed toS. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 strains with and without rat liver S9 activation byplate incorporation assay at the concentration of0, 1, 2, 3, 4, 5 and 5 mg/plate. DMSO was used as solvent. Negative and positive control were used .Anisyl Formate 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 rat liver S9 activation byplate incorporation assay .Although positive response is observed in some strains, there were also no apparent trends discernible that would suggest a change in mutagenic activity caused by the addition of the ingredient chemicals. Therefore Anisyl Formate(122-91-8) was not likely to classify as gene mutant in vitro.

Supported by a experimental study conducted by Richard R. Baker et al.(Food and Chemical Toxicology ,2004), Gene mutation toxicity study in vitro was performed to determine the mutagenic nature of Anisyl Formate. Genetic toxicity in vitro study was assessed for Anisyl Formate (122-91-8) to evaluate its possible mutagenic potential .For this purpose AMES assay was performed according to guideline OECD 471.Anisyl Formate were tested inSalmonella typhimuriumstrains TA98, TA100, TA102, TA1535 and TA1537 with and without S9 metabolic activation by Bacterial gene mutation assay. Statistically significant positive responses with revertant response-NFDPM dose relationships were observed in both control and treated bacterial cell. No mutagenic effects were observed in any of the treated strain. Therefore, Anisyl Formatewas considered to be negative gene toxic. 

It is further supported by experimental study conducted by Richard R. Baker et al.(Food and Chemical Toxicology ,2004), Gene mutation toxicity study in vitro was performed to determine the mutagenic nature of Anisyl Formate using mammalian cell line. Genetic toxicity in vitro study was assessed for Anisyl Formate (122-91-8) to evaluate its possible mutagenic potential .For this purpose micronucleus assay was performed according to guideline OECD 473.Anisyl Formate were tested by using V79 Chinese hamster micronucleus cell line with and without S9 metabolic activation. Cigarette smoke TPM produced from the test cigarettes did not produce any biologically-relevant increases in the levels of micronucleus induction, above those of the relevant control cigarette TPM. The test material was exposed to the cell line at the concentration of 0, 5, 10, 15, 20, 25, 30 and 35 µg/ml .Therefore; Anisyl Formatewas considered being negative gene toxic.  

Based on the data and prediction, available for the target chemical Anisyl Formate IUPAC : 4-methoxyphenyl)methyl formate (122-91-8) does not induce gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.

Justification for classification or non-classification

Thus based on the above annotation for the target chemical Anisyl Formate IUPAC : 4-methoxyphenyl)methyl formate (122-91-8)  and CLP criteria is not likely to classify as a gene mutant in vitro.