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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
Study period:
1997
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Deviations:
no
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Diacetone alcohol
- Physical state: Colorless transparent liquid
- Analytical purity: 99.8%
- Storage conditions: Stored in a cool dark location

Method

Target gene:
Histidine opero
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Species / strain / cell type:
E. coli WP2 uvr A
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Phenobarbital and 5,6-benzoflavone-induced rat liver microsomes (S9)
Test concentrations with justification for top dose:
0, 313, 625, 1250, 2500, or 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Japanese Pharmacopeia water for injection
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
Japanese Pharmacopeia water for injection
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: - See Table 1
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 minutes at 37°C
- Exposure duration: 48 hours at 37°C

NUMBER OF REPLICATIONS: 3 plates for both the solvent and the positive control groups in the dose range finding tests and 1 for each dose. Additionally, during this study, 3 plates were used for each of the control groups and each dose. Independent repeat performed.
METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation; in suspension; as impregnation on paper disk

DETERMINATION OF CYTOTOXICITY
Dose levels: 0, 50, 150, 500, 1500, or 5000 µg/plate (±S9)

Evaluation criteria:
Of the five types of assay strains, under the conditions with and without the S9 mix in at least one type of assay strain, if the mean values for the number of mutant colonies on a plate containing the test substance were more than double than those of the solvent control when verifying the reproducibility and dose dependence, the test substance was deemed to be mutagenic (positive) for this study. If only one of these two tests confirmed a dose at which the mean number of colonies was double or more that of the solvent control values, and if the solvent control value was 10 or less, dose dependency could not be confirmed by an increase in the number of mutant colonies, it was deemed to be negative.
Statistics:
None required (number of revertant colonies counted).

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:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative
Executive summary:

DAA was tested a bacterial reverse mutation assay (according to OECD guideline 471) at doses of 0, 313, 625, 1,250, 2,500, or 5,000 µg/plate in Salmonella typhimurium strains TA 98, TA 100, TA 1535, and TA 1537 and Escherichia coli strain WP2 uvr A both in the absence and presence of exogenous metabolic activation (phenobarbital and 5,6-benzoflavone-induced rat liver S9).  Incubations at each concentration were done in triplicate and an independent repeat experiment was performed. Water for injection was used as the vehicle and positive controls were included in all incubations. No cytotoxicity and no increase in the reverse mutation rate were observed at any DAA concentration in any of the tester strains either in the absence or presence of metabolic activation. Incubation with positive control substances either in the absence or presence of metabolic activation resulted in anticipated increases in the reverse mutation rate.