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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:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed publication

Data source

Reference
Reference Type:
publication
Title:
Gene mutaton toxicity study of the test chemical
Author:
Haworth et al
Year:
1983
Bibliographic source:
Environmental Mutagenesis

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
The Salmonella/mammalian microsome test was performed to determine the mutagenic nature of the test chemical in vitro
GLP compliance:
not specified
Type of assay:
bacterial gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: 7-Diethylamino-4-methylcoumarin
- Molecular formula: C14H17NO2
- Molecular weight: 231.2933 g/mol
- Substance type: Organic
- Physical state: Powder Solid
- Purity: Label purity: Practical
Analyzed purity: No data available
- Impurities (identity and concentrations): No data available

Method

Target gene:
Histidine
Species / strain
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
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
Liver S-9 fractions were prepared from male Sprague-Dawley rats and male Syrian hamsters
Test concentrations with justification for top dose:
0, 100, 333, 1000, 3333, 5450 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Soluble and stable in DMSO
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 2-Aminoanthracene, 4-Nitro-o-phenylenediamine
Details on test system and experimental conditions:
METHOD OF APPLICATION: Preincubation

DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 h
- Expression time (cells in growth medium): 48 h
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: Three plates were used, and the experiment was repeated no less than 1 week after completion of the initial test.

NUMBER OF CELLS EVALUATED: No data available

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available

OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available

OTHER: No data available
Rationale for test conditions:
No data
Evaluation criteria:
Increase in the number of revertants
Statistics:
Statistical analysis of Salmonella plate test data was performed as per Margolin et al, 1981

Results and discussion

Test results
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:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
True negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data available
- Effects of osmolality: No data available
- Evaporation from medium: No data available
- Water solubility: No data available
- Precipitation: No data available
- Other confounding effects: No data available

RANGE-FINDING/SCREENING STUDIES: To select the dose range for the mutagenesis assay, the test chemicals were checked for toxicity to TA100 up to a concentration of 10 mg/plate or the limit of solubility, both in the presence and absence of S-9 mix. One or more parameters were used as an indication of toxicity: viability on complete medium (EGG) and reduced numbers of revertant colonies per plate and/or thinning or absence of the bacterial lawn (CWR, EGG, SRI). If toxicity was not apparent in the preliminary toxicity determination, the highest dose tested was 10 mg/plate; otherwise the upper limit of solubility was used. If toxicity was observed, the doses of test chemical were chosen so that the high dose exhibited some degree of toxicity.

COMPARISON WITH HISTORICAL CONTROL DATA: No data available
Remarks on result:
other: No mutagenic potential

Any other information on results incl. tables

Dose(µg/plate)

TA100

NA

RLI

HLI

0.0(Solvent control)

114±10.5

151±13.9

132±5.2

100.0

118±5.9

148±15.2

115±6.1

333.0

108±3.3

121±8.8

113±2.3

1000.0

93±3.5

107±2.5

130±4.6

3333.0

127±10.9

140±4.4

127±18.0

5450.0

122±12.3

141±9.6

150±21.5

POS

506±21.0

698±47.9

1522±90.7

Dose(µg/plate)

TA1535

NA

RLI

HLI

0.0(Solvent control)

6±2.2

12±0.6

5±1.2

100.0

7±1.2

8±1.9

6±1.9

333.0

4±0.3

7±0.6

7±2.0

1000.0

4±1.8

5±1.2

3±1.5

3333.0

2±1.5

7±1.3

3±1.5

5450.0

3±0.3

5±1.7

2±0.6

POS

345±6.2

143±40.0

77±4.3

Dose(µg/plate)

TA1537

NA

RLI

HLI

0.0(Solvent control)

4±1.5

8±0.0

10±1.5

100.0

5±0.9

6±0.6

11±3.2

333.0

5±1.0

11±2.6

7±1.5

1000.0

5±1.2

9±2.0

8±2.6

3333.0

3±0.7

9±1.9

8±3.2

5450.0

3±1.0

6±2.6

7±0.9

POS

829±8.5

60±8.2

69±2.7

Dose(µg/plate)

TA98

NA

RLI

HLI

0.0(Solvent control)

16±2.0

21±1.7

23±1.2

100.0

16±0.9

16±2.0

23±1.8

333.0

13±0.7

17±2.0

21±4.7

1000.0

15±2.0

22±5.9

18±3.8

3333.0

15±4.1

23±2.3

20±4.6

5450.0

9±1.2

20±2.7

22±3.4

POS

348±29.0

414±29.4

1263±155.7

RLI: rat liver S-9 Aroclor 1254 induced;

HLI: hamster liver S-9Aroclor 1254 induced,

Mutagenic responses of Salmonella tester strains TA100, TA1535, TA1537, and TA98 (mean ± SEM) to test chemicals.

Applicant's summary and conclusion

Conclusions:
The test chemical is not mutagenic to Salmonella typhimurium TA100, TA1535, TA1537, TA98 in the preincubation assay performed with and without S9 metabolic activation system and hence does not classify as a gene mutant in vitro.
Executive summary:

The Salmonella/mammalian microsome test was performed to determine the mutagenic nature of test chemical in vitro.

 

Preincubation assay was performed using Salmonella typhimurium TA100, TA1535, TA1537, TA98 with and without S9 metabolic activation system. To select the dose range for the mutagenesis assay, the test chemicals were checked for toxicity to TA100 up to a concentration of 10 mg/plate or the limit of solubility, both in the presence and absence of S-9 mix.

 

The doses thus selected were 0, 100, 333, 1000, 3333, 5450 µg/plate. Appropriate positive controls were also incorporated in the study.

The test chemical is not mutagenic to Salmonella typhimurium TA100, TA1535, TA1537, TA98 in the preincubation assay performed with and without S9 metabolic activation system and hence does not classify as a gene mutant in vitro.