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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:
1980
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1980
Report Date:
1980

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
Identity FAT 65004/F TE
Batch 15060201 (China)
Purity determined in this study
Appearance
Smell
yellowish powder, solid at 20°C
neutral
pH-Value pH-value of a soln. of 2% (w/w) = 6.90
Expiration date August 25th, 2020
Storage to be stored at room-temperature
Specific details on test material used for the study:
None

Method

Target gene:
histidine-auxotrophic strains of Salmonella typhimurium
Species / strain
Species / strain / cell type:
other: histidine- auxotrophic TA 98, TA 100, TA 1535, TA 1537 and TA 1538, strains of Salmonella typhimurium and Strain E. coli WP2 uvrA
Metabolic activation:
with and without
Metabolic activation system:
S9 fraction of liver from rats
Test concentrations with justification for top dose:
5, 10, 50, 100, 500, 1000 and 5000 µg/0.1 ml.
Vehicle / solvent:
Dimethylsulfoxide (DMSO)
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Details on test system and experimental conditions:
The tests were performed with the following concentrations of the trial substance without and with microsomal activation: 5, 10, 50, 100, 500, 1000 and 5000 µg/0.1 ml.

Preparation of test solution:
Solvent : Dimethylsulfoxide (DMSO); Merck
Amount per plate: 0.1 ml
Method for suspension of refractory substances: Ultrasonication
Time of preparation: On the day of the experiment

Media:
(1) Minimal agar Composition: Vogel-Bonner Medium E, 1,5% Agar, 2% Glucose
(2) Soft agar
Amount per plate: 2 ml
Temperature: 45°C
Composition: 0.6% agar +0.6 % NaCl in twice distilled water 400 ml + 40ml solution of histidine (0.5mM)/biotin (0.5mM) (with S. typhimurium) or 40 ml solution of tryptophan (0.5mM) (with E. coli)


Test method and conditions:
(1) Test Method: Plate Method
(2) Test conditions
Composition of the plates
Bacterial suspension: 0.1 ml/plate
Test solution : 0.1 ml/plate
Na phosphate buffered solution: -
S9 Mix where required: 0.5 ml/plate
Soft agar solution, 45°C: 2.0 ml/plate
Others Conditions:
Incubation temperature: 37°C
Incubation time: 48 - 55 hours

The tests were carried out in accordance with the method described by AMES et al
Rationale for test conditions:
None
Evaluation criteria:
The criteria of mutagenicity used in this test are a doubling of the spontaneous reversion rate and a dose-effect relationship.
Statistics:
None

Results and discussion

Test results
Key result
Species / strain:
other: strains of Salmonella typhimurium: TA 98, TA l00, TA 1535, TA 1537, TA 1538 and E. coli WP2 uvrA
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
None

Any other information on results incl. tables

None

Applicant's summary and conclusion

Conclusions:
It is concluded that FAT65004/D is not mutagenic to histidine-auxotrophic strains of Salmonella typhimurium: TA 98, TA l00, TA 1535, TA 1537, TA 1538 and E. coli WP2 uvrA
Executive summary:

FAT 65 004/D was tested for mutagenic effects on histidine-auxotrophicstrains of Salmonella typhimurium and on a tryptophan-auxotrophic strain of E. coli. The investigations were performed with the following concentrations of the trial substance with and without microsomal activation: 5, 10, 50, 100, 500, 1000 and 5000 µg/0.1 ml.

 

These tests permit the detection of point mutations in bacteria induced by chemical substances. Any mutagenic effects of the substances are demonstrable on comparison of the number of bacteriain the treated and control cultures that have undergone back-mutationto histidine- or tryptophan-prototrophism. To ensurethat mutagenic effects of metabolites of the test substanceformed in mammals would also be detected, experiments were performedin which the cultures were additionally treated with anactivation mixture (rat liver microsomes and co-factors).

In the experiment performed without and with microsomal activation,comparison of the numbers of back-mutant colonies in the controls and the cultures treated with the various concentrations of FAT 65004/D revealed no marked deviations.  

No evidence of the induction of point mutations by FAT 65004/D or by the metabolites of the substance formed as a result ofmicrosomal activation was detectable in the strains of S. typhimuriumand E. coli used in these experiments.

It is concluded that FAT65004/D is not mutagenic to  histidine-auxotrophic strains of Salmonella typhimurium: TA 98, TA l00, TA 1535, TA 1537, TA 1538 and E. coli WP2 uvrA