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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: Guideline study. GLP accordance not stated in the report. Test article precipitated in soft agar from the lowest concentration used in the Ames assay. No analytical monitoring of test concentration was done.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1986
Report date:
1986

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
(1981)
Deviations:
yes
Remarks:
no statistical analysis was performed
GLP compliance:
no
Remarks:
not stated in the report
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Details on test material:
- Purity: Commercial grade
- Lot/batch No.: Op. 11006
- Stability of test article: In the report it was stated that stability was guaranteed by the sponsor.


Method

Target gene:
All strains are histidine auxotrophic mutants.
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):
not applicable
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium TA 102
Details on mammalian cell type (if applicable):
not applicable
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
One millilitre activation mixture contained 0.3 ml S9 fraction of liver from rats (Tif:RAIf(SPF)) induced with Aroclor 1254 and 0.7 ml of a solution of co-factors.
Test concentrations with justification for top dose:
A preliminary toxicity test was carried out with concentrations ranging from 0.08 to 5000 µg/plate suspended in dimethylsulfoxide.
Ames test (without and with microsomal activation): 20, 78, 313, 1250 and 5000 µg/plate suspended in dimethylsulfoxide.
Vehicle / solvent:
- Vehicle used: dimethylsulfoxide (DMSO)
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
vehicle DMSO
True negative controls:
no
Positive controls:
yes
Remarks:
Without S9-Mix
Positive control substance:
other: Daunorubicin-HCl (5 and 10 µg/plate)
Remarks:
TA 98
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
vehicle DMSO
True negative controls:
no
Positive controls:
yes
Remarks:
Without S9-Mix
Positive control substance:
4-nitroquinoline-N-oxide
Remarks:
TA 100

Migrated to IUCLID6: 0.125 and 0.25 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
vehicle DMSO
True negative controls:
no
Positive controls:
yes
Remarks:
Without S9-Mix
Positive control substance:
mitomycin C
Remarks:
TA 102

Migrated to IUCLID6: 0.5 and 1 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
vehicle DMSO
True negative controls:
no
Positive controls:
yes
Remarks:
Without S9-Mix
Positive control substance:
sodium azide
Remarks:
TA 1535

Migrated to IUCLID6: 2.5 and 5 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
vehicle DMSO
True negative controls:
no
Positive controls:
yes
Remarks:
Without S9-Mix
Positive control substance:
9-aminoacridine
Remarks:
TA 1537

Migrated to IUCLID6: 50 and 100 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
vehicle DMSO
True negative controls:
no
Positive controls:
yes
Remarks:
With S9-Mix
Positive control substance:
other: 2-aminoanthracene 5 µg/plate
Remarks:
TA 98, TA 100, TA 1537
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
vehicle DMSO
True negative controls:
no
Positive controls:
yes
Remarks:
With S9-Mix
Positive control substance:
other: 2-aminoanthracene 20 µg/plate
Remarks:
TA 102
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
vehicle DMSO
True negative controls:
no
Positive controls:
yes
Remarks:
With S9-Mix
Positive control substance:
cyclophosphamide
Remarks:
TA 1535

Migrated to IUCLID6: 250 µg/plate
Details on test system and experimental conditions:
The study consisted of a toxicity pre-screen test followed by the Ames assay (with and without S9-Mix).

OTHER EXAMINATIONS
A preliminary toxicity test was carried out with the concentrations ranging from 0.08 to 5000 µg/plate. Thereafter, the Ames test was performed with 20, 78, 313, 1250 and 5000 µg/plate test article (with and without microsomal activation). The substance precipitated in soft agar at and above a concentration of 20 µg/plate.

MUTATION TEST
Mutation assays with and without microsomal activation were performed twice in triplicate per strain and per group (standard plate test).
Each Petri dish contained approximately 20 ml of minimum agar, 0.1 ml of the test substance or the vehicle and 0.1 ml of a bacterial culture (in nutrient broth) in 2.0 ml of soft agar. Experiments with microsomal activation contained additionally 0.5 ml of an activation mixture, consistent of S9 fraction of liver from Aroclor 1254 induced rats and co-factors. The plates were incubated for about 48 hours at 37 ± 1.5°C in darkness.

EVALUATION CRITERIA
Arithmetic mean of revertant colonies was calculated. The test substance is considered to be non-mutagenic if the colony count in relation to the negative control is not doubled at any concentration.
Statistics:
No statistical analysis

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:
other: Cytotoxicity was assessed in a pretest up to 5000 µg/plate. No further details are available.
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: Cytotoxicity was assessed in a pretest up to 5000 µg/plate. No further details are available.
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: The substance precipitated in soft agar at and above a concentration of 20 µg/plate.

Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

The number of histidine-prototrophic mutants after treatment with the substance did not vary markedly in comparison to the negative control in any of the experiments.

Applicant's summary and conclusion

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