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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:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report Date:
2000

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes
Type of assay:
bacterial gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Purity test date: 99.85%
- Lot/batch No.: 98033005
- Expiration date of the lot/batch: 30 july 2000
-storage condition: at room temperature and protected from light.

Method

Target gene:
Five strain of bacteria salmonella typhimurium: TA 1535, TA1537, TA98, TA 100 and TA 102.
Species / strain
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:

The concentrations are:
.312,5; 625; 1250; 2500 and 5000 µg/plate for the strain 102 in the first assay.
. 156.25; 312.5; 625; 1250; and 2500 µg/plate



Vehicle:
Dimethyldisulfide (DMSO)
Controls
Negative controls:
no
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
benzo(a)pyrene
Remarks:
Migrated to IUCLID6: sodium azide, 9-Aminoacridine, 2-Nitrofluorene, Mytomicin
Details on test system and conditions:
All experiment were performed according to the direct plate incorporation method except for the second and third tests with S9 mix, which were perfomed according to the preincubation method (60 minutes, 37°C).
Evaluation criteria:
The evaluation of the toxicity was performed on the basis of the observation of the decrease in the number of revertant colonies and/or a thinning of the bacterial lawn.
Statistics:
not applicable

Results and discussion

Test results
Species / strain:
other: TA1535, TA 1537, TA98, TA100 et TA102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
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

Under our experimental conditions, the test susbtance n-heptanol (batch No. 9803005, purity: 99.5%) does not show mutagenic activity in the bacterial reverse mutation test with salmonella typhimurium.
Executive summary:

The objective of this study was to evaluate the potential of the test substance  n-HEPTANO L (batchNo.9803005,purity:99.85%)  to induce reverse mutation in Salmonella typhimurium.

 

  A preliminary toxicity test was performed to define the dose-levels of n-HEPTANOL  to be used for the mutagenicity study . The test substance was then tested in two independent experiments, with and without a metabolic activation system, the S9mix, prepared from alive rmicrosomal fraction(S9fraction) of rats induced with Aroclor1254. A third experiment was performed with S9mix.

 

All experiments were performed according to the direct plate incorporation method except for the second and third tests with S9 mix,which were performed according to the preincubation method(60minutes,37°C).

 

Five strains of bacteria Salmonella typhimurium:TA1535, TA1537, TA98, TA lOO and TA102 were used. Each  strain  was exposed  to five  dose-levels  of the test substance (three plates/dose-levet).After 48 to 72 hours of incubation at 37°C,the revertant colonies were scored. The evaluation of the toxicity was performed on the basis of the observation of the decrease in the number of revertant colonies and/or a thinning of the bacterial lawn.

 

The test substance n-HEPTANOL  was dissolved in dimethylsulfoxide  (DMSO). The dose-levels of the positive controls were as follows :

without S9 mix:

g/plateofsodiumazide(NaN3):TA1535andTA100strains,

50µg/plate of 9-Aminoacridine(9AA):TA1537strain,

0.5 µg/plate of 2-Nitrofluorene(2NF):TA98strain,

.0.5 µg/plate of Mitomycin C (MMC):TA102strain.

 

with S9 mix:

. 2 µg/plate of 2-Anthramine (2AM): TA1535,TA1537,TA98andTAlOOstrains,

. 10µg/plate of 2-Anthramine (2AM): TA102strain.

 

 

 

Results

 

Since the test substance was toxic in the preliminary test, the choice of the highest dose-level for the main test was based

 on the level of toxicity, according to the criteria specified in the international guidelines.


 

Experiments without S9 mix:

The selected treatment-levels were as follows:

.312.5,625,1250,2500 and 5000 µg/plate, for the TA102 strain in the first experiment,

.156.25,312.5,625,1250 and 2500 µg/plate, for the all tester strains except for the TA102 strain in the first experiment as well as for the TA98 and TA102 strains in the second experiment,

. 78.125,156.25, 312.5,625 and 1250 µg/plate, for the TA1535,TA1537 and TA100 strains in the second experiment.

 

A slight to strong toxicity was noted,depending on the tester strain and the dose-levels.

 

The test substance did not induce any noteworthy increase in the number of revertants,in both experiments,in any of the five strains.

 

Experiments with S9 mix:

The selected treatment-levels were as follows:

. 312.5,625,1250, 2500 and 5000 µg/plate, for the TA102strain in the first experiment,

.156.25,312.5,625, 1250 and 2500µg/plate, for the all tester strains except for the TA102 strain in the first experiment as well as for the TA1537 and TA 102 strains in the second experiment,

78.125,156.25,312.5,625 and 1250 µg/plate, for the TA1535, TA98 and TA 100 strains in

these cond experiment as weil as for the TAl02 strain in the third experiment,

39.06,78.125,156.25,312.5and 625µg/plate, for all tested strains,except for the TA102 strain, in the third experiment.

 

A slight to strong toxicity was induced,depending on the tester strain,the dose-level and the experimental conditions.

 

The test substance did not induce any noteworthy increase in the number of revertants, in all the experiments, in any of the five strains.

 

The number of revertants for the vehicle and positive controls was as specified in the acceptance criteria. The study

  was therefore considered valid.

 

 Under our experimental conditions, the test substance n-heptanol (batch N. 9803005, purity: 99.5% ) does not show mutagenic activity in the bacterial reverse mutation test with salmonella typhimurium.