Registration Dossier

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:
1989
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study without detailed documentation.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report Date:
1989

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
reliability scoring based on 1997 guideline.
Deviations:
yes
Remarks:
Salmonella typhimurium TA102 or Escherichia coli-WP2 uvrA strain not tested; 2-aminoanthracene was the only compound used to test the efficacy of the S9 fraction.
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: colourless liquid
Details on test material:
- Name of test material (as cited in study report): Sec-butylchlorid
- Physical state: Liquid
- Analytical purity: Not reported
- Storage condition of test material: 4°C

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Rat liver S9 (compound used to induce CYP450 not reported)
Test concentrations with justification for top dose:
8, 40, 200, 1000 and 5000 µg/plate

Vehicle / solvent:
- Vehicle(s)/solvent(s) used: ethanol
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
ethanol
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: see Table 1
Details on test system and experimental conditions:
METHOD OF APPLICATION: plate incorporation

DURATION
- Exposure duration: 48 hours at 37 °C

NUMBER OF REPLICATIONS: 4 plates per strain and dose; two independent experiments performed.


Evaluation criteria:
A reproducible and dose related increase in the number of mutant counts for at least one strain is considered positive. For TA 100, TA 98 and TA 1535 a two fold increase of revertants compared to the negative controls should be reached, whereas for TA 1537 a threefold increase should be reached. Otherwise the results are deemed to be negative.
Statistics:
Revertant colony numbers were calculated as average and standard deviation. No statistical analysis was performed.

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:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Remarks:
(5000 µg/plate)
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES WITH STRAIN TA100:
Test concentration: 19.5, 39, 78, 156, 312, 625, 1250, 2500, 5000 and 10000 µg/plate
No bacterial toxicity was observed.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

There was no indication of a bacteriotoxic effect of Sec-butylchloride in all of the employed doses. The total bacteria counts consistently produced results comparable to the negative controls (i.e. vehicle controls). An inhibition of bacterial growth was not noted. 

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

No evidence of mutagenic activity of sec-butylchlorid was found.
Executive summary:

No evidence of mutagenic activity of sec-butylchlorid was found. In this study four strains (TA98, TA100, TA1535 and TA1537) were tested with and without metabolic activation (S9 -mix) in concentrations up to 5000 µg/plate.