D-Glucitol, 1,3:2,4-bis-O-[(4-chlorophenyl)methylene]-

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007-08-08 - 2008-04-03

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: non-GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

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:

chromosome aberration assay

Test material

Test material information

Constituent 1

Reference substance name:

(1R)-1-[(4R,4aR,8aS)-2,6-bis(4-chlorophenyl)-hexahydro-[1,3]dioxino[5,4-d][1,3]dioxin-4-yl]ethane-1,2-diol

EC Number:

700-591-1

Cas Number:

82203-23-4

Molecular formula:

C20H20Cl2O6

IUPAC Name:

(1R)-1-[(4R,4aR,8aS)-2,6-bis(4-chlorophenyl)-hexahydro-[1,3]dioxino[5,4-d][1,3]dioxin-4-yl]ethane-1,2-diol

Details on test material:

- Name of test material (as cited in study report): DCBS
- Substance type: organic
- Physical state: white powder

Test animals

Species:

mouse

Strain:

other: Kun-ming Mouse, SPF

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: no data
- Weight at study initiation: 25—30g
- Assigned to test groups randomly: Yes
- Fasting period before study: no data
- Housing: Male and female mice were fed in separate cages (L46 X W31 xH20cm). The cages were changed twice each week.

- Diet (e.g. ad libitum):
Feed was offered freely throughout the study period.

- Water (e.g. ad libitum): Water was offered freely throughout the study period.
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ±3 °C
- Humidity (%): 30—70 %
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark.

IN-LIFE DATES: From: To: no data

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: vegetable oil
- Justification for choice of solvent/vehicle: no data
- Concentration of test material in vehicle: 5000 mg, 2500mg, 1250 mg / 40 mL
- Amount of vehicle (if gavage or dermal): 40 mL

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
According to the doses design, 5000 mg, 2500 mg and 1250 mg of testing article were taken and then was-added with vegetable oil to 40 mL, misce bene for test.
The three doses of reagents, positive control (cyclophosphamide, 40mg/kg) and negative control (vegetable oil 20mL/kg) were administrated intragastrically respectively to the five groups, at a proportion of 0.4mL/20g weight. Animals from each group are sacrificed at 24, 36 and 48 hours after administration.

Duration of treatment / exposure:

24, 36 and 48 hours

Frequency of treatment:

gavage in one time

Post exposure period:

Animals from each group were sacrificed at 24, 36 and 48 hours after administration

Doses / concentrations

Remarks:

Doses / Concentrations:
5000 mg/kg, 2500mg/kg, 1250mg/kg
Basis:
nominal conc.

No. of animals per sex per dose:

15 male and 15 female

Control animals:

yes, concurrent vehicle

Positive control(s):

cyclophosphamide
- Justification for choice of positive control(s): no data
- Route of administration: gavage
- Doses / concentrations: 40mg/kg

Examinations

Tissues and cell types examined:

Bone marrow from femora

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: no data

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields):
Bone marrow is removed from femora of all animals and then made into slides with calf serum.

DETAILS OF SLIDE PREPARATION:
The slide is prepared, fixed and stained with Giemsa

METHOD OF ANALYSIS:
Record the number of PCEs(polychromatic erytrocytes) which contain micronucleus of the 2000 PCEs per mouse. Calculate the permil of the cells containing micronucleus. Count the 200 PCE as well and the NCE, calculating the PCE/NCE ratio.

Evaluation criteria:

No data

Statistics:

Using chi-square criterion to analyze the micronucleus percent of each group with negative control group statistically

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

other: Negative control is the vehicle control

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- no preliminary test recorded

RESULTS OF DEFINITIVE STUDY
- the ratio of Micronucleus and PCE/ NCE of test group have no obvious difference comparing with negative control (vehicle control) group. And the positive control show obvious difference.
- The mouse weight change of the test group has no obvious differece comparing with negative control (vehicle control) group and positive control.

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative
No toxic effect of the test item was observed in the experimental conditions, therefore, the genetic toxicity of test item is considered to negative.