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Genetic toxicity: in vitro

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in vitro gene mutation study in bacteria
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Does not meet important criteria of today's standard methods, methodological deficiencies in Ames experiment.

Data source

Reference Type:
study report
Report Date:

Materials and methods

Principles of method if other than guideline:
Overlay method Ames test as in Ames et al., Mutation Research 31: 347, 1975.
GLP compliance:
Type of assay:
bacterial reverse mutation assay

Test material

Details on test material:
- Description: Colourless liquid
- Analytical purity: not stated


Target gene:
Histidine synthesis gene
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9 fraction
Test concentrations with justification for top dose:
0.001 - 5.0 ul/plate equivalent to 1.5 - 7565 ug/plate
Vehicle / solvent:
Solvent was either deionised water of dimethylsulfoxide (DMSO); used to prepare stock solutions of solid materials. All dilutions of test materials were made in either deionised water or DMSO. The solvent employed and its concentration are recorded in the results section.
Negative solvent / vehicle controls:
Deionised water or DMSO
Positive controls:
MNNG, NF, QM were used as nonactivation controls and ANTH, AAF and AMQ were used as activation controls
Details on test system and experimental conditions:
Approximately 10(8) cells from an overnight culture of each indicator strain were added to seperate test tubes containing 2ml of molten agar supplemented with biotin and a trace of histidine .For non-activation tests, at least four dose levels of the test compound were added to the contents of the appropriate tubes and poured over the surface of selective agar plates. In activation tests, a minimum of four different concentrations of the test chemical were added to the appropriate tubes with cells. Just prior to pouring, an aliquot of reaction mixture was added to each of the activation overlay tubes, which were then mixed and the contents poured over the surface of a minimal agar plate and allowed to solidify. The plates were incubated for 48hours at 37 degrees centigrade and scored for the number of colonies growing on each plate. The concentrations of all chemicals are given in the results section. Positive and solvent controls using both directly active positive chemicals and those that require metabolic activation were run with each assay.
Evaluation criteria:
The numbers of colonies in each plate were counted and recorded on printed forms. These raw data were analysed in a computer program and reported in a printout.

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
TDCP did not produce any increase in the number of revertants.
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
The test compund did not demonstrate mutagenic activity in any of the assays conducted in this evaluation and was considered as not mutagenic under these test conditions.
Remarks on result:
other: all strains/cell types tested
Migrated from field 'Test system'.

Applicant's summary and conclusion

Interpretation of results (migrated information):