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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:
key study
Study period:
April 1983
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1983
Report Date:
1983

Materials and methods

Test guidelineopen allclose all
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
OECD Guidelines for testing of chemicals, Draft No. 419 (status 1983)
Qualifier:
equivalent or similar to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
yes
Remarks:
EEC Directive 79/831, Annex V, Method No. 431 (status 1983)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
- Physical state: see above
- Appearance: see above
Specific details on test material used for the study:
TEST MATERIAL:
- Identification: P 5256
- Batch/Lot number: not available
- Purity: not available
- Physical appearance: powder
- Stability: pure: not available / in vehicle: not available
- Storage: the test material was stored at room temperature in the dark
To avoid any light effects on the test compound, all experimentation was performed under yellow light

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
Bacterial test organisms: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537
Source: B.N. Ames , University of California, Berkeley CA, 94720, USA, May 1981

Characterization of strains:
The strains used are mutants derived from Salm. typhimurium LT2 and have following genotype:
- S.typhimurium TA 1535 his G46 r fa- uvrB
- S.typhimurium TA 1537 his C3076 r fa- uvrB
- S.typhimurium TA 98 his D3052 r fa- uvrB- R+
- S.typhimurium TA 100 his G46 r fa- uvrB- R+
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium TA 1538
Details on mammalian cell type (if applicable):
Bacterial test organisms: Salmonella typhimurium TA 1538
Source: B.N. Ames , University of California, Berkeley CA, 94720, USA, May 1981

Characterization of strains:
The strains used are mutants derived from Salmonella typhimurium LT2 and have following genotype: S.typhimurium TA 1538 his D3052 r fa- uvrB
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 liver fractions of rats, treated with AROCLOR 1254
Test concentrations with justification for top dose:
1.58 , 5, 15.8, 50, 158, 500, 1580, 5000 micrograms per plate; substance was not toxic up to the highest concentration

Vehicle / solvent:
DMSO
Controls
Untreated negative controls:
yes
Remarks:
DMSO
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
N-ethyl-N-nitro-N-nitrosoguanidine
methylmethanesulfonate
Details on test system and experimental conditions:
Method of application: in agar (plate incorporation)
Evaluation criteria:
A material is identified as a mutagen in this test system if there is a reproducible demonstration of a dose effect relation with a 2-fold increase in the number of revertants over the controls in a minimum of one strain. With the strain TA 100, a 1.5-fold increase is the criterion for a positive result.
Statistics:
no data

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid

Applicant's summary and conclusion

Conclusions:
It can be stated that during this in vitro Salmonella/mammalian-microsome assay with P 5256, no gene mutagenic activity could be demonstrated under the experimental conditions reported.
Executive summary:

This study was performed to investigate the potential gene mutagenic activity of the test item according to the plate incorporation test of Ames et. al. (1) using the Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538. The test was performed with and without liver microsomal activation. The test material was tested at the following concentrations: 1.58, 5, 15.8, 50, 158, 500, 1580 and 5000 micrograms per plate. Each concentration, including the controls, was tested in triplicate. No toxic effect of the test material was observed. Up to the highest investigated dose, no relevant increase of the revertant colony numbers was obtained in any Salmonella typhimurium strain when compared with the corresponding controls. The presence of microsomal activation did not influence these findings. In conclusion, it can be stated that during this in vitro Salmonella/mammalian-microsome assay, no gene mutagenic activity could be demonstrated under the experimental conditions reported.