1,2-diacetoxybut-3-ene

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

no

GLP compliance:

yes

Type of assay:

other: Mouse micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

1,2-diacetoxybut-3-ene

EC Number:

421-720-5

EC Name:

1,2-diacetoxybut-3-ene

Cas Number:

18085-02-4

Molecular formula:

C8H12O4

IUPAC Name:

1-(acetyloxy)but-3-en-2-yl acetate

Test material form:

liquid

Test animals

Species:

mouse

Strain:

CD-1

Details on species / strain selection:

Crlj: CD1(ICR) strain

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: CRL Japan
- Age at study initiation: 7 weeks
- Weight at study initiation: 32.9 to 36.8 g (male DRF), 25.4 to 29.3 g (female DRF), 34 to 36.8 g (male micronucleus test).
- Assigned to test groups randomly: Yes
- Housing: Polycarbonate cages (group housed)
- Diet (e.g. ad libitum): Pelleted diet (adlib)
- Water (e.g. ad libitum): Provided adlib
- Acclimation period: At least 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3°C
- Humidity (%): 50 +/- 20°C
- Air changes (per hr): > 10 changes per hour
- Photoperiod (hrs dark / hrs light): 12 h day (150 to 300 Lux)

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: Olive oil
- Justification for choice of solvent/vehicle: Substance insoluble in water and 0.5% carboxymethyl cellulose.
- Concentration of test material in vehicle: 10 mL/Kg

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The test substance 4000 mg and 1000 mg were weighed for the tests of dose determination and micronucleus assay respectively. These were added to a volume of olive oil and adjusted to 20 mL (DRF) or 5 mL (micronucleus test). The lower doses were serially diluted from the concentration. Test solutions were prepared before use.

Duration of treatment / exposure:

DRF: 24 or 48 hours
Micronucleus test: 24 hours

Frequency of treatment:

Administered once

Post exposure period:

24 or 48 hours (DRF) and 24 hours (micronucleus test)

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

125 mg/kg bw/day (nominal)

Remarks:

DRF

Doses / concentrations 2

Dose / conc.:

250 mg/kg bw/day (nominal)

Remarks:

DRF

Doses / concentrations 3

Dose / conc.:

500 mg/kg bw/day (nominal)

Remarks:

DRF

Doses / concentrations 4

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

DRF

Doses / concentrations 5

Dose / conc.:

2 000 mg/kg bw/day (nominal)

Remarks:

DRF

Doses / concentrations 6

Dose / conc.:

500 mg/kg bw/day (nominal)

Remarks:

Micronucleus assay

Doses / concentrations 7

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

Micronucleus assay

Doses / concentrations 8

Dose / conc.:

2 000 mg/kg bw/day (nominal)

Remarks:

Micronucleus assay

No. of animals per sex per dose:

2 males and 2 females per dose group in the DRF and 5 males per dose group in the micronucleus study.

Control animals:

yes, concurrent vehicle

Positive control(s):

Positive control: Mitomycin C (as prescribed in the test guideline)

Examinations

Tissues and cell types examined:

Bone marrow cells

Details of tissue and slide preparation:

The animals were sacrificed by cervical dislocation at 24 or 48 hours after dosing in the DRF and after 24 hours in the micronucleus test. The right femurs were dissected and bone marrow cells were treated and prepared on to glass slides using standard procedures. The slides were subsequently stained as appropriate.

Evaluation criteria:

Positive: The number of MNPCE increased dose-dependantly or significantly increased at a single dose point compared with negative controls.
Negative: Not meeting positive criteria.

Statistics:

The incidences of MNPCE and the ratio of PCE to RBC of each test substance concentration group and positive control group were compared with that of the negative control group for significant differences using Wilcoxon's rank sun test.

Results and discussion

Test results

Sex:

male

Genotoxicity:

negative

Toxicity:

yes

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

The induced frequency of the MNPCE in the positive control group was significantly higher with that of the negative control group. The PEC ratio to RBC did not differ compared with the negative control group. The results show that the study was conducted appropriately, and all values values were within relevant historical ranges.

Applicant's summary and conclusion

Conclusions:

The experimental findings conclude that the test substance was negative for mutagenicity as the substance did not increase the incidence of MNPCE in the mouse femoral bone marrow.