Octan-4-olide

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 13th 200 - September 13th 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 guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Octan-4-olide

EC Number:

203-208-1

EC Name:

Octan-4-olide

Cas Number:

104-50-7

Molecular formula:

C8H14O2

IUPAC Name:

octan-4-olide

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Haarmann & Reimer GmbH; 50529270
- Purity: 99.2%

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

Aroclor-1254 indiuced rat liver S9

Test concentrations with justification for top dose:

Main test 1: 0, 15, 50, 150, 500, 1500, 5000 µg/plate
Main test 2: 0, 15, 50, 150, 500, 1500, 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

mitomycin C

other: 2-Aminoanthracene: All strains +S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation) for both experiments

DURATION
- Exposure duration: 48 – 72 hrs

NUMBER OF REPLICATIONS: 2

DETERMINATION OF CYTOTOXICITY
- Method: background lawn reduction

Statistics:

Estimation of the statistical significance of the difference between the mean number of revertants in the negative controls and the plates at each dosage level, using a Chi squared (X2) test.

Results and discussion

Test results

Test results 1

Species / strain:

other: S. typhimurium TA 1537, TA 98 and TA 102

Remarks:

Main test 1 & 2

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

5000 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

other: S. typhimurium TA 1535, TA 100

Remarks:

Main test 1 & 2

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Species / strain:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100

Remarks:

Main tests 1 & 2

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 102

Remarks:

Main tests 1 & 2

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

5000 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation of the test compound an the plates was not observed.

RANGE-FINDING/SCREENING STUDIES: According to an initial toxicity test, OCTALACTONE GAMMA was tested in concentrations of 15 to 5000 lag per plate in the presence and absence of S9 (data not shown).

HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%)
- Positive historical control data: A historical overview of the revertant frequencies of the strains used in the Freiburger Labor für Mutagenitätsprüfung of the years 1998 to 2000 was provded for positive controls
- Negative historical control data: A historical overview of the revertant frequencies of the strains used in the Freiburger Labor für Mutagenitätsprüfung of the years 1998 to 2000 was provided for spontaneous and solvent controls.

Applicant's summary and conclusion

Conclusions:

In conclusion, these results indicate that octan-4-olide, under the experimental conditions described, was not mutagenic to Salmonella typhimurium strains TA1535, TA1537, TA98, TA100, and TA102 in the presence and absence of a metabolizing system.

Executive summary:

In a reverse gene mutation assay in bacteria (2000073), strains of S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 1022 were exposed to octan-4-olide (99.2%) in DMSO at concentrations of 0, 15, 50, 150, 500, 1500, 5000 µg/plate (plate incorporation, both experiments) in the presence and absence of mammalian metabolic activation (Aroclor-1254 induced rat liver S9).

Octan-4-olide was tested up to limit concentrations (5000 µg/plate). The positive controls induced the appropriate responses in the corresponding strains. There was no evidence of induced mutant colonies over background.

This study is classified as acceptable. This study satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data.