1,3-Oxathiolane-2-carboxylic acid, 5-hydroxy-, (1R,2S,5R)-5-methyl-2-(1-methylethyl)cyclohexyl ester, (2R,5R)-

Genetic toxicity: in vitro

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:

11.08.2004 to 12.10.2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed according to the OECD guideline 471 and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(1R,2S,5R)-2-isopropyl-5-methylcyclohexyl (2R,5R)-5-hydroxy-1,3-oxathiolane-2-carboxylate

EC Number:

604-569-1

Cas Number:

147126-62-3

Molecular formula:

C14H24O4S

IUPAC Name:

(1R,2S,5R)-2-isopropyl-5-methylcyclohexyl (2R,5R)-5-hydroxy-1,3-oxathiolane-2-carboxylate

Constituent 2

Reference substance name:

OS Menthol Ester

IUPAC Name:

OS Menthol Ester

Constituent 3

Reference substance name:

(2R, 5R)-5-Hydroxy-[1,3] oxathiolane-2-carboxy1ic acid, 2S-isopropyl-5Rcmethyl-1R-cyclohexyl ester

IUPAC Name:

(2R, 5R)-5-Hydroxy-[1,3] oxathiolane-2-carboxy1ic acid, 2S-isopropyl-5Rcmethyl-1R-cyclohexyl ester

Test material form:

other: White to light pink/yellow solid

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Additional strain / cell type characteristics:

other: Tryptophan-deficient

Metabolic activation:

with and without

Metabolic activation system:

S9 mix (rat)

Test concentrations with justification for top dose:

Experiment I: s. typhimurium: 0, 7.4, 22, 67, 200 and 600 µg/plate and 0, 0.82, 2.47, 74, 22 and 67µg/plate with and without S9, respectively
E. coli: 0, 22, 67, 200, 600 and 1800 µg/plate and 0, 0.82, 2.47, 7.4, 22, 67 µg/plate with and without S9, respectively
Experiment II: 0, 0.82, 2.47, 74, 22 and 67µg/plate and 0, 0.27, 0.82, 2.47, 7.4 and 22 µg/plate with and without S9, respectively
E. coli: 0, 22, 67, 200, 600 and 1800 µg/plate and 0, 0.82, 2.47, 7.4, 22, 67 µg/plate with and without S9, respectively

Vehicle / solvent:

DMSO

Controls

Controls 1

Positive controls:

yes

Remarks:

5.0µg/plate

Positive control substance:

2-nitrofluorene

Remarks:

TA98 without metabolic activation

Controls 2

Positive controls:

yes

Remarks:

5.0µg/plate

Remarks:

TA100 without metabolic activation

Controls 3

Positive controls:

yes

Remarks:

5.0µg/plate

Positive control substance:

2-nitrofluorene

Remarks:

TA1535 without metabolic activation

Controls 4

Positive controls:

yes

Remarks:

100µg/plate

Positive control substance:

9-aminoacridine

Remarks:

TA1537 without metabolic activation

Controls 5

Positive controls:

yes

Remarks:

1.0µL/plate

Remarks:

WP2 uvrA without metabolic activation

Controls 6

Positive controls:

yes

Remarks:

5.0µg/plate

Positive control substance:

benzo(a)pyrene

Remarks:

TA98 with metabolic activation

Controls 7

Positive controls:

yes

Remarks:

5.0µg/plate

Positive control substance:

benzo(a)pyrene

Remarks:

TA100 with metabolic activation

Controls 8

Positive controls:

yes

Remarks:

100µg/plate

Positive control substance:

cyclophosphamide

Remarks:

TA1535 with metabolic activation

Controls 9

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

TA1537 with metabolic activation

Controls 10

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Details on test system and experimental conditions:

METHOD OF APPLICATION: 1st main experiment: in agar (plate incorporation); 2nd main experiment: preincubation
DURATION
- 1st main experiment: 48 - 72 hours incubation (37°C)
- 2nd main experiment: 30 min preincubation (30°C), 48 - 72 hours incubation (37°C)
NUMBER OF REPLICATIONS: 3
DETERMINATION OF CYTOTOXICITY
- Method: determination of background lawn

Evaluation criteria:

Positive result: A concentration-related increase over the range tested and/or a reproducible increase in at least one or more concentrations in the number of revertant colonies per plate in at least one strain with or without metabolic activation system. Biological relevance ofthe results will be considered first. A
statistical method may be used as an aid in evaluating the test results but it should not be the only determining factor. A positive result indicates that the test substance induces point mutations in S. typhimurium or E. coli.

Negative result: If a result does not meet the above criteria, it will be reported as negative. A negative result indicates that the test substance is non-mutagenicc in this test. Equivocal result: If no definite judgement can be made to fit the above criteria, even after repeated experiments, then the result will be described as equivocal.

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

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

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 with metabolic activation
negative without metabolic activation

OS Menthol Ester was not mutagenic to S.typhimurium strains, TA98, TA100 TA1535, TA1537, and E.coli strain, WP2 uvrA, under the conditions of
the test.

Executive summary:

Based on the results of this study the substance is considered to be not mutagenic to S. Typhimurim strains T98, T100, T1535 and T1537 as well as E. colit strain WP2 uvrA with and without metabolic activation.