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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:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
26Feb 1990 - 16Mar 1990
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
This study has been performed according to OECD and/or EC guidelines and according to GLP principles. Deviation: Only Salmonella typhimurium strains were tested, no E coli-strain was included. In repeat experiment only 3 instead of 5 analysable concentrations were tested.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report Date:
1990

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
No E.coli-strain was included. In repeat experiment only 3 instead of 5 analysable concentrations were tested.
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
Annex V of EEC Commission Directive 84/449/EEC
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Substance: colourless liquid
- Batch number: 3-2
- Storage conditions: in the dark
- Purity: not reported in this report

Method

Target gene:
Histidine gene
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
other: uvrB and rfa deficient
Species / strain / cell type:
S. typhimurium TA 1538
Additional strain / cell type characteristics:
other: uvrB and rfa deficient
Metabolic activation:
with and without
Metabolic activation system:
S9-mix (Aroclor 1254 induced)
Test concentrations with justification for top dose:
Preliminary test:
0; 312.5; 625; 1250; 2500; 5000 µg/ plate
Main study:
0; 8; 40; 200; 1000; 5000 µg/ plate (experiment 1)
0; 312.5; 625; 1250; 2500; 5000 µg/ plate (experiment 2)
Vehicle / solvent:
- DMSO
- Justification for choice of solvent/vehicle: DMSO control plates gave counts of revertant colonies within the normal range.
Controlsopen allclose all
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
other: N-Methyl-N'-nitro-N-nitrosoguanidine at 2 µg/ plate for TA1535 and TA100; 9-Aminoacridine at 50 µg/ plate for TA1537; 4-Nitro-O-phenylenediamine at 10 µg/ plate for TA1538 and TA98
Remarks:
without S9
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene at 2.5µg/ plate
Remarks:
with S9
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation);

CYTOTOXICITY: Growth of background lawn

NUMBER OF REPLICATIONS: 2 (preliminary) or 3 (main study)
Evaluation criteria:
- A substance is considered positive if a dose-related and statistically significant increase in mutation rate in one or more strains of bacteria in the presence and/or absence of S9-mix in both experiments at sub-toxic dose levels is found.
- A substance is considered negative if the number of induced revertants compared to spontaneous revertants is less than twofold at each dose level employed, the intervals of which should be between 2 and 5 fold and extend to the limits imposed by toxicity, solubility or up to 5000 µg/ plate.
Statistics:
All data are statistically analysed using the methods recommended by UKEMS (Kirkland (Ed.): Statistical Evaluation of Mutagenecity Test Data", UKEMS Sub-committee on guidelines for Mutagenicity Testing. Report - Part III (1989)- Cambridge university Press).

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:
cytotoxicity
Remarks:
≥ 1250 µg/ plate
Vehicle controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
≥ 1250 µg/ plate
Vehicle controls validity:
valid
Positive controls validity:
valid

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

In an AMES test according to OECD/EC guidelines under GLP circumstances using 5 different Salmonella typhimurium strains with two independent repeats, Nisseki-SAS-40 was found not to be mutagenic with or without metabolic activation.
Executive summary:

SAS-40 was tested in the Salmonella typhimurium reverse mutation assay according to OECD/EC guidelines with five histidine-requiring strains of Salmonella typhimurium (TA1535, TA1537, TA1538, TA98 and TA100) with and without metabolic activation (S9 -mix). The test was performed in two independent experiments. The test substance did not induce a significant dose-related increase in the number of revertant (His+) kolonies in any of the strains. Both the solvent control and positive controls gave results within expected ranges. Cytotoxicity was observed at ≥ 1250 µg/ plate. Based on the results of this study it can be concluded that SAS-40 is not mutagenic in the Salmonella typhimurium reverse mutation assay.