[No public or meaningful name is available]

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

Study period:

29 June 2004 to 16 July 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 474 (Mammalian Erythrocyte Micronucleus Test)

Version / remarks:

1997

Deviations:

no

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Test material form:

liquid

Test animals

Species:

mouse

Strain:

ICR

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Orient C., Ltd., Seoul, Korea
- Age at study initiation: six weeks
- Weight at study initiation: 23.5-27.0 g
- Assigned to test groups randomly: using algorithm of Path/Tox so that each group had similar body weight distribution
- Fasting period before study:
- Housing: up to six animals per cage in polycarbonate cages with bedding
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23°C ± 3°C
- Humidity (%): 50%
- Air changes (per hr): 10-20
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: mid-May 2004 To: July 8 2004

Administration / exposure

Route of administration:

oral: unspecified

Vehicle:

- Vehicle(s)/solvent(s) used: olive oil
- Justification for choice of solvent/vehicle: test item is insoluble in water
- Concentration of test material in vehicle: no information
- Amount of vehicle (if gavage or dermal): no information. It is not stated if administration was by gavage, but this is presumed to be the case

Duration of treatment / exposure:

24 hours

Frequency of treatment:

Two treatments at 24 hour intervals

Doses / concentrations

Remarks:

Doses / Concentrations:
500, 1000 and 2000 mg/kg bw
Basis:

No. of animals per sex per dose:

Six males

Control animals:

yes, concurrent vehicle

Positive control(s):

Positive control substance: cyclophosphamide
- Justification for choice of positive control(s): widely used in micronucleus assays
- Route of administration: intraperitoneal
- Doses / concentrations: 70 mg/kg bw

Examinations

Tissues and cell types examined:

Bone marrow

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: preliminary toxicity study

TREATMENT AND SAMPLING TIMES (in addition to information in specific fields): animals were sacrificed 24 hours after the second administration

DETAILS OF SLIDE PREPARATION: cells were collected in fetal bovine serum, centrifuged and smeared onto slides. Preparations were dried and fixed by submerging in methanol. Slides were stained with May-Grunwald and Giesma stains, then rinsed dried and mounted.

METHOD OF ANALYSIS: Slides were examined under x1000 magnification. Small round or oval bodies (1/5 to 1/20 diameter of PCE) were counted as micronuclei. Micronuclei were counted in 200 PCE's. The (PCE):(PCE+NCE) ratio was calculated by counting 500 cells

Evaluation criteria:

The result was judged as positive if there was a statistically significant and dose-related increase or a reproducible increase in the frequency of MNPCEs at least at one dose level.

Statistics:

Differences of numbers of MNPCRs between treated and vehicle control: Kruskal-Wallis' H-test and Dunnett's test.
Differences of numbers of MNPCRs between positive and vehicle control: Mann-Whitney's U-test.
Differences of PCE/(PCE+NCE) between treated and vehicle control:

Results and discussion

Test results

Key result

Sex:

male

Genotoxicity:

negative

Toxicity:

no effects

Remarks:

No effects observed in bone marrow, and no clinical signs were noted

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Any other information on results incl. tables

Table 1: Micronucleus test in male ICR mice

Chemical treated	Dose (mg/kg)	No. of animals	MNPCE/2000 PCEs	PCE/(PCE+NCE)
Vehicle	0	6	1.17	0.52
Test item	500	6	1.33	0.49
Test item	1000	6	1.83	0.55
Test item	2000	6	2.00	0.55
CPA	70	6	94.83**	0.45*

* Significantly different from the control at P <0.01

** Significantly different from the control at P < 0.05

Applicant's summary and conclusion

Conclusions:

The test substance has been tested in an in vivo micronucleus assay conducted according to OECD 474 and in compliance with GLP. No evidence of induction of micronuclei was observed in male ICR mice. No signs of systemic toxicity were observed and the test item did not cause toxicity in the bone marrow. Solvent and positive controls gave expected responses. It is concluded that the test substance is negative for the induction of micronuclei under the conditions of the study.