[No public or meaningful name is available]

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 February 1998

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Version / remarks:

Adopted July 1997

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Remarks:

quality assurance statement included

Type of assay:

other: Micronucleus assay in rats

Test material

Test material information

Constituent 1

Reference substance name:

Chondroitin disaccharide deltadi-4S sodium salt

Cas Number:

136144-56-4

Molecular formula:

C14H19NO14S•2Na

IUPAC Name:

Chondroitin disaccharide deltadi-4S sodium salt

Constituent 2

Reference substance name:

α-deltaUA-[1→3]-GalNAc-6S = α-[4-deoxy-L-threo-hex-4-enopyranosyluronic acid] sodium salt-[1→3]-N-acetyl-D-galactosamine-6-sulphate

Cas Number:

136132-72-4

Molecular formula:

C14H19NO14S•2Na

IUPAC Name:

α-deltaUA-[1→3]-GalNAc-6S = α-[4-deoxy-L-threo-hex-4-enopyranosyluronic acid] sodium salt-[1→3]-N-acetyl-D-galactosamine-6-sulphate

Constituent 3

Reference substance name:

unknown metal constituents

IUPAC Name:

unknown metal constituents

Constituent 4

Reference substance name:

Chloride

Cas Number:

16887-00-6

Molecular formula:

Cl

IUPAC Name:

Chloride

Constituent 5

Reference substance name:

Ethanol

EC Number:

200-578-6

EC Name:

Ethanol

Cas Number:

64-17-5

Molecular formula:

C2H6O

IUPAC Name:

Ethanol

Constituent 6

Reference substance name:

unknown constituent

Molecular formula:

unknown

IUPAC Name:

unknown constituent

Constituent 7

Reference substance name:

Glucuronic acid

EC Number:

229-486-4

EC Name:

Glucuronic acid

Cas Number:

6556-12-3

Molecular formula:

C6H10O7

IUPAC Name:

(2S,3S,4S,5R,6R)-3,4,5,6-Tetrahydroxyoxane-2-carboxylic acid / (2S,3S,4S,5R,6R)-3,4,5,6- Tetrahydroxyoxane-2-carboxylic acid

Constituent 8

Reference substance name:

(2R,3S,5R)-3-amino-6-(hydroxyméthyl)oxane-2,4,5-triol

Cas Number:

7535-00-4

Molecular formula:

C6H13NO5

IUPAC Name:

(2R,3S,5R)-3-amino-6-(hydroxyméthyl)oxane-2,4,5-triol

Constituent 9

Reference substance name:

group of unknown constituents

Molecular formula:

unknown

IUPAC Name:

group of unknown constituents

Test material form:

solid

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

162 g to 190 g (males) 137 g to 164 g (females), 5 to 6 weeks old
Environmental conditions: period of acclimatization of mini 5 days; ventilated rooms; temperature of 22°C and humidity of 55%; lighting 12 hours a day.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

1% Carboxymethylcellulose in distilled water

Details on exposure:

oral intubation

Frequency of treatment:

3 daily treatments

Post exposure period:

24h

Doses / concentrations

Dose / conc.:

2 000 mg/kg bw/day

Remarks:

dose administered three times

No. of animals per sex per dose:

2 males and 2 females, exposed to 3 consecutive doses in a preliminary assay, 5 males and 5 females, exposed to 3 consecutive doses of the test substance in a confirmatory assay,

Control animals:

yes

Positive control(s):

Exposed to cyclophosphamide, a clastogenic agent, at dose of 25 mg/kg (intraperitoneal route)

Examinations

Tissues and cell types examined:

Erythrocytes

Details of tissue and slide preparation:

The bone marrow is extracted from the rats femurs with foetal calf serum. The cell suspensions are centrifuged for 5 min at 1000 rpm. The supernatant is eliminated and the centrifugate is spread on slides. The smears are stained using a technique derived from the May Grunwald Giemsa technique which makes it possible to distinguish between polychromatic and normochromatic erythrocytes.

Evaluation criteria:

Ratio between normochromatic and polychromatic erythrocytes, determined by analysis of 1000 erythrocytes/animal min.

Statistics:

Statistical analysis is performed for micronucleus numbers using the Mann Whitney U rank test (non parametric test used because the distribution of the numbers of micronuclei does not correspond to a Gaussian distribution but to a Poisson-type distribution).
The statistical comparison for the polychromatic/normochromatic erythrocyte ratio and for the weight homogeneity within the sex of each group is performed using the Student's test.

Results and discussion

Test results

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Remarks:

mortality endpoint only

Vehicle controls validity:

valid

Negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

The validity criteria were fulfilled.
The minimum lethal dose of the test compound was higher than 2000 mg/kg/d (x3) by the oral route in rat. Under these conditions this dose was retained as the limit dose for the micronucleus test according to the OECD guideline recommendation.
There was no statistically significant difference between the weights of trated and control rats.

Applicant's summary and conclusion

Conclusions:

In an OECD 474 micronucleus test in male and female rats, no mortality nor clastogenic activity can be demonstrated after 3 administrations of the test substance by oral route at 2000 mg/kg/d.