Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
Data is from publication.

Data source

Reference
Reference Type:
publication
Title:
The genotoxic potential of nicotine and its major metabolites
Author:
Doolittle, D. J., Winegar, R., Lee, C. K., Caldwell, W. S., Hayes, A. W. and deBethizy, J. D
Year:
1995
Bibliographic source:
Mutat. Res., 1995, 344, 95-102,1995

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:
To evaluate the mutagenic potential of test chemical in Salmonella Typhimurium strain TA 1535, TA 1537, TA 98 and TA 100 by Salmonella/microsome Assay.
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 (as cited in study report): Nicotine
- Substance type: organic
- Physical state: liquid
- Analytical purity: nicotine > 99.5% (determined by GC-MS)

Method

Target gene:
Histidine
Species / strain
Species / strain:
other: TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell lines (if applicable):
Not specified
Additional strain characteristics:
not specified
Cytokinesis block (if used):
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 mix (5%) from male Sprague-Dawley rats given a single 500 mg/kg injection i.p. of Aroclor 1254
Test concentrations with justification for top dose:
0 ,62.5,125.0,250.0,500.0, 750.0 and 1000.0 µg/plate
Vehicle:
phosphate buffer
Controls
Negative controls:
yes
Solvent controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: 9-aminoacridine 2-nitrofluorene sodium azide 2-aminoanthracene
Details on test system and conditions:
METHOD OF APPLICATION: Preincubation
DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h
- Triplicate plates at each concentration
Rationale for test conditions:
Not specified
Evaluation criteria:
A sample was considered to be mutagenic if it induced a concentration-dependent increase in revertants number with at least one concentration being at least two times the solvent control.
Statistics:
Yes ,Mean standard deviation was observed

Results and discussion

Test results
Species / strain:
other: TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
not specified
Negative controls valid:
not specified
Positive controls valid:
yes
Remarks on result:
other: No mutagenic effect were observed.
Additional information on results:
Not specified

Applicant's summary and conclusion

Conclusions:
Test chemical was evaluated for its mutagenic potential in Salmonella typhimurium TA 1535, TA 1537, TA 98 and TA 100 by AMES test. The test result was considered to be negative in all strain in the presence and absence of metabolic activation S9.
Executive summary:

Genetic toxicity in vitro study was assessed for test chemical. The test material was exposed to Salmonella typhimurium TA 1535, TA 1537, TA 98 and TA 100 in the presence and absence of metabolic activation S9, and 20 min standard Preincubation time. The test chemical was exposed at the concentration of 0, 62.5, 125, 250, 500, 750 and 1000 µg/plate. No mutagenic effects were observed in all strains, in the presence and absence of metabolic activation. Therefore test chemical was considered to be non mutagenic in Salmonella typhimurium TA 1535, TA 1537, TA 98 and TA 100 by AMES test. Hence the substance cannot be classified as gene mutant in vitro.