REAKTIV OLIV F00-149

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:

2002-02-27 to 2002-04-29

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5395 (In Vivo Mammalian Cytogenetics Tests: Erythrocyte Micronucleus Assay)

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

442-640-7

EC Name:

-

Cas Number:

479541-17-8

Molecular formula:

Hill formula: C32H26CuFN9Na4O19S5

IUPAC Name:

Tetrasodium 2-(4-fluoro-6-(2-(2-sulfonatoethansulfonyl)ethyl amino)-1,3,5-triazin-2-ylamino)-5-hydroxy-6-(2-(2-hydroxy-5- sulfonatophenylazo)-4,5-dimethoxyphenylazo)-7-sulfonatonaphthalene-1-sulfonatocuprate(II)

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): Reaktiv Oliv F00-0149

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Winkelmann GmbH Gartenstrasse 27, 33178 Borchen
- Age at study initiation: 6 weeks
- Weight at study initiation: mean male: 190.6 g; mean female: mean= 149.9 g
- Housing: five animals per cage in transparent macrolon cages (type IV) on soft wood granulate in an air-conditioned room (room number 011)
- Diet (e.g. ad libitum): rat/mice diet ssniff R/M-H (V 1534), ad libitum
- Water (e.g. ad libitum): tap water in plastic bottles, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22
- Humidity (%): 50
- Photoperiod (hrs dark / hrs light): 12h light/dark

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

deionized water

Duration of treatment / exposure:

24 h

Frequency of treatment:

two doses separated by an interval of 24 hours (except positive control with only one dose)

Doses / concentrations

Remarks:

Doses / Concentrations:
2000 mg/kg
Basis:
no data

No. of animals per sex per dose:

15

Control animals:

yes, concurrent vehicle

Positive control(s):

The positive control was with Endoxan, which was administered once orally at a dose of 40 mg per kg body weight.

Examinations

Tissues and cell types examined:

bone marrow: 2000 polychromatic erythrocytes were counted for each animal

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION:
Based on the results of the acute oral toxicity-study PTOl-0361 the dose of 2000 mg/kg bw was selected as the limit dose

DETAILS OF SLIDE PREPARATION:
bone marrow smaples were fentrifuged with fetal bovine serum, sediment was smeared and stained as follows:
Staining procedure
-5 minutes in methanol
-5 minutes in May-Griinwald' s solution
-brief rinsing twice in distilled water
-10 minutes staining in 1 part Giemsa solution to 6 parts buffer solution, pH 7.2 (Weise) rinsing in distilled water
-drying
-coating with Entellan®

METHOD OF ANALYSIS:
2000 polychromatic erythrocytes were counted for each animal. The number of cells with micronuclei was recorded

Evaluation criteria:

Criteria for a positive response:
A substance is considered as positive if there is a dose-related increase in the number of micronucleated polychromatic erythrocytes or a significant increase in at least one dose group compared with the concurrent negative control group which is above the range of the historical control data. A test substance producing no significant or dose-related increase in the number of micronucleated polychromatic erythrocytes is considered non-clastogenic in this system.

Statistics:

Assuming the study is valid based on a monotone-dose-relationship, one-sided Wilcoxon tests were performed initially comparing control values with those of the highest dose group. A significance level of 5% is adopted for all tests.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

There was a slight increase in micronucleated polychromatic erythrocytes in male animals, but was within the historical control range of the negative control groups and was concluded to have no biological relevance.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative
Reactive Olive F00-0149 did not cause a substantial increase of micronucleated polychromatic erythrocytes and is therefore not considered clastogenic in the micronucleus test in vivo.

Executive summary:

In a Sprague-Dawley rat bone marrow micronucleus assay, 15 rats/sex were treated orally with two doses, 24 hours apart, of Reactive Olive at 2000 mg/kg bw. Bone marrow cells were harvested 24h post-treatment. The vehicle was deionized water. 

There were no signs of substance-related toxicity during the study. The positive control induced the appropriate response. There was not a significant increase in the frequency of micronucleated polychromatic erythrocytes in bone marrow after treatment.

This study is classified asacceptable and satisfies the requirement for Test Guideline OPPTS 870.5395; OECD 474 for in vivo cytogenetic mutagenicity data.