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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non-GLP, non-guideline study, published in peer reviewed literature, minor restrictions in design and/or reporting but otherwise adequate for assessment
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
publication
Title:
Improved method for mutagenicity testing of gaseous compounds by using a gas sampling bag
Author:
Araki A, Noguchi T, Kato F & Matsushima T
Year:
1994
Bibliographic source:
Mutation Research, 307, 335-344

Materials and methods

Test guideline
Qualifier:
no guideline followed
Deviations:
not applicable
Principles of method if other than guideline:
Mutagenicity test on S. Typhimurium TA98, TA100, TA1535 and TA1537 and E. Coli WP2 uvrA using the developed gas exposure method (using a gas sampling bag as an exposure vessel and a preparation vessel of diluted gas).
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
2-butene was obtained from Tokyo Kasei Co. Ltd, Tokyo, Japan

Method

Target gene:
not applicable
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
E. coli WP2 uvr A
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 from Phenobarbitol and 5,6-benzoflavone induced rat liver
Test concentrations with justification for top dose:
Toxic concentration levels or about 50% of the maximum exposure concentration.
Vehicle / solvent:
Vehicle(s)/solvent(s): none (diluted by HEPA-filtered air)
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
not specified
True negative controls:
not specified
Positive controls:
not specified
Positive control substance:
not specified
Remarks:
The objective was to develop an improved method for testing gases. Use of standard positive and negative control substances was not stated, buta,-1,3-diene was used as a positive substance & the data presented indicates that an air control was used
Details on test system and experimental conditions:
Mutagenicity testing was performed by the gas exposure method using a gas sampling bag with or without metabolic activation. The exposure condition was : bacterial plates made by the agar overlay method with 2mL top agar per plate, 100 µL of S9 per plate, exposure temperature of 37°C, exposure period of 48 h and exposure volume of gas was 500 mL per plate. Tests were performed at toxic concentration levels or about 50% levels as the maximum concentration.

The conditions listed above were chosen based on the results of a preliminary study with 1,3 butadiene in which the effect of the volume of gas per plate (357, 625, 1250, 2500 or 5000 mL/plate, which corresponded to 14, 8, 4, 2 or 1 plates/bag), the amount of S9 (50, 100, 200 or 400 µL/plate), the exposure temperature (30 or 37°C), exposure period (2, 4, 14, 24 or 48 hours), the amount of top agar (0, 1, or 2 mL/plate) were examined.
Evaluation criteria:
After incubation the number of revertant colonies was counted.
Statistics:
none

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
none
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

none

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

2 -Butene was not mutagenic, with and without metabolic activation, in S. Typhimurium strains: TA 1535, TA 1537, TA 98 and TA 100 and E. Coli WP2 uvr A.
Executive summary:

2-butene was not mutagenic to S Typhimurium TA98, TA100, TA1535, TA 1537 and E.coli WP2uvrA, with or without metabolic activation, using a gas sampling bag method. Concentrations were not specified but tests were carried out at increasing levels until toxicity was observed. Buta-1,3-diene was mutagenic under these conditions indicating that the method was robust.