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Toxicological information

Genetic toxicity: in vitro

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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:
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

Reference
Name:
Unnamed
Type:
Constituent
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:
other: TA97, TA98, TA100, TA102, TA104, TA2638 and UTH8413 and UTH8414
Details on mammalian cell lines (if applicable):
Not applicable
Additional strain 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:
- Vehicle(s)/solvent(s) used: Sterile double distilled water
- Justification for choice of solvent/vehicle: Chemical solubility in Sterile double distilled water
Controls
Negative controls:
not specified
Solvent 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 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:
other: TA97, TA98, TA100, TA102, TA104, TA2638 and UTH8413 and UTH8414
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not specified
Positive controls valid:
yes
Remarks on result:
other: No mutagenic potential
Additional information on results:
No data

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.