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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 Aug 2000 to 5 Oct 2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to guideline
Guideline:
other: EEC-Guideline 92/69 L 383 A, B.14
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
cis-2-tert-butylcyclohexyl acetate
EC Number:
243-718-1
EC Name:
cis-2-tert-butylcyclohexyl acetate
Cas Number:
20298-69-5
Molecular formula:
C12H22O2
IUPAC Name:
2-tert-butylcyclohexyl acetate
Test material form:
liquid
Details on test material:
Test material used is consistent with the substance identity as described in section 1.2 of IUCLID - company composition for the quality described under the common name Verdox.

Method

Target gene:
his
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA97a, Ta98, TA100, TA102 and TA1535
Metabolic activation:
with and without
Metabolic activation system:
rat S9
Test concentrations with justification for top dose:
Study 1:
Strains TA971, TA100, TA1535, -S9: 0.016, 0.05, 0.16, 0.5 and 1.6 mg/plate
Strains TA97a (+S9), TA98 (+/-S9), TA100 (+S9), TA1535 (+S9), TA102 (+/-S9): 0.05, 0.16, 0.5, 1.6 and 5 mg/plate

Study 2:
Strains TA97 and TA1535 (-S9): 0.016, 0.05, 0.16, 0.5 and 2.6 mg/plate
Strains TA97 (+ S9), TA98 (+/-S9), TA100 (+/-S9), TA1535 (+S9) and TA102 (+/-S9): 0.05, 0.16, 0.5, 1.6 and 5 mg/plate.
Vehicle / solvent:
- Vehicle used: DMSO
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: ICR 191, 4-nitro-1,2-phenylenediamine, nitrofurantoine, natrium azide, 2-aminoanthracene, cumene hydroperoxide, danthron.
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Incubation period: 18 hours
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 3 replicates per concentration level and control

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth
Evaluation criteria:
For evaluation of the results the induction rate of the mean values was calculated: induction rate = (revertant colonies of the test item)/(revertant colonies of the corresponding control). The test item is to be interpretated mutagenic if there is a concentration effect relationship and the induction rate is ≥ 2.

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 97a
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES: The range of test concentrations was determined in a preliminary (non-GLP) test with the concentration rage 0.05-5 mg/plate (factor 10).

ADDITIONAL INFORMATION ON CYTOTOXICITY: the lowest cytotoxic concentrations were 1.6 mg/plate for TA97a (-S9), 5 mg/plate for TA100 (+/- S9), 1.6 mg/plate for TA1535 (-S9) and 5 mg/plate for TA1535 (+ S9). No cytotoxicity was observed in strains TA97a (+ S9), TA98 (+/-S9) and TA102 (+/-S9).
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
The substance gave negative results in the Ames test with Salmonella typhimurium strains TA97a, TA98, TA100, TA102 and TA1535, with and without metabolic activation, in a concentration range of 0.016-5 mg/plate.
Executive summary:

The ability of the test substance to induce gene mutations in Ames test was tested in the GLP-compliant OECD Guideline 471 study in Salmonella typhimurium strains TA97a, TA98, TA100, TA102 and TA1535, with and without metabolic activation. The tested concentration ranges were 0.05 -5 mg/plate for strains TA97a (+S9), TA98 (+/-S9), TA100 (+S9), TA102 (+/-S9) and TA1535 (+S9), and 0.016 -1.6 mg/plate for TA97a (-S9), TA100 (-S9) and TA1535 (-S9). The lowest cytotoxic concentrations were 1.6 mg/plate for TA97a (-S9) and TA1535 (-S9), and 5 mg/plate for TA 100 (+/-S9) and TA1535 (+ S9). No cytotoxicity was observed for strains TA97a (+S9), TA98 (+/-S9) and TA102 (+/-S9). The tests were performed in triplicate. No increased number of revertants was observed in any case. Based on the results of the study, the substance is concluded to be negative in Amest test.