1,4-dimethyl-5-[[4-[methylbenzylamino]phenyl]azo]-1H-1,2,4-triazolium acetate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

Mutagenicity and Cytotoxicity of N-Methyl-2-pyrrdidinone and 4-(Methy1amino)butanoic Acid in the Salmonella/Microsome Assay

Author:

David A. Wells, Harvey F. Thomas and George A. Digenis

Year:

1988

Bibliographic source:

Journal of Applied Toxicology, Vol. 8(2), 135-139 (1988)

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

Ames assay was performed to determine the mutagenic nature of n-methyl-2-pyrrolodone

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1-mehyl-2-pyrrolodone

Cas Number:

872-50-4

Molecular formula:

C5H9NO

IUPAC Name:

1-mehyl-2-pyrrolodone

Details on test material:

- Name of test material: n-methyl-2-pyrrolodone
- IUPAC name: 1-methyl-2-pyrrolodone
- Molecular formula: C5H9NO
- Molecular weight: 99.1321 g/mol
- Substance type: Organic

Specific details on test material used for the study:

- Name of test material: n-methyl-2-pyrrolodone
- IUPAC name: 1-methyl-2-pyrrolodone
- Molecular formula: C5H9NO
- Molecular weight: 99.1321 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA97, TA98, TA100, TA102, TA104, TA2638 and UTH8413 and UTH8414

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

S9 liver homogenates were prepared from Aroclor induced rats

Test concentrations with justification for top dose:

0, 0.01, 0.1, 1.0, 10.0, 100.0 or 1000.0 µmol/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Sterile double distilled water
- Justification for choice of solvent/vehicle: Chemical solubility in Sterile double distilled water

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Sterile double distilled water

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

other: please refer remarks

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: No data
- Exposure duration: 2 days
- Expression time (cells in growth medium): 2 days
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: In all experiments four plates for each concentration of test chemical were used and each experiment was repeated once.

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

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

OTHER: No data

Rationale for test conditions:

No data

Evaluation criteria:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: No data
- Exposure duration: 2 days
- Expression time (cells in growth medium): 2 days
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: In all experiments four plates for each concentration of test chemical were used and each experiment was repeated once.

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

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

OTHER: No data

Statistics:

Pooled data were analyzed by Bonferroni t-tests of differences between means of the treatment doses vs control, at a significance level of P = 0.05. All calculations were performed by computer using theStatistical Analysis System (SAS Institute Inc., Cary. NC).

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA97, TA98, TA100, TA102, TA104, TA2638 and UTH8413 and UTH8414

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

Additional information on results:

No data

Remarks on result:

other: No mutagenic potential

Applicant's summary and conclusion

Conclusions:

N-methyl-2-pyrrolodone did not induce gene mutation in Salmonella typhimurium TA97, TA98, TA100, TA102, TA104, TA2638 and UTH8413 and UTH8414 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

Executive summary:

Ames assay was performed to determine the mutagenic nature of N-methyl-2-pyrrolodone. The study was performed as per the method described by Ames et al using Salmonella typhimurium TA97, TA98, TA100, TA102, TA104, TA2638 and UTH8413 and UTH8414 with and without S9 metabolic activation system. The test chemical was dissolved in sterile double distilled water depending on solubility and at dose levels of0, 0.01, 0.1, 1.0, 10.0, 100.0 or 1000.0µmol/plate. Concurrent solvent and positive control chemicals were included in the study. The plates were observed for a dose dependent and a 2-fold increase in the number of revertants/plate. All assay procedures were performed under yellow light to avoid photodynamic effects. N-methyl-2-pyrrolodonedid not induce gene mutation in Salmonella typhimurium TA97, TA98, TA100, TA102, TA104, TA2638 and UTH8413 and UTH8414 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.