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Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Type of information:
experimental study
Adequacy of study:
key study
Study period:
July 1st, 1996 to July 18th, 1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report date:
1996

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.5395 (In Vivo Mammalian Cytogenetics Tests: Erythrocyte Micronucleus Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Version / remarks:
1983
Deviations:
no
GLP compliance:
yes
Type of assay:
mammalian erythrocyte micronucleus test

Test material

Constituent 1
Reference substance name:
3‐benzoyl‐2,6‐bis({[3‐(‐lambda5‐diazynylidene)‐4‐oxo‐3,4‐dihydronaphthalen‐1‐yl]sulfonyl}oxy)phenyl 3‐(‐lambda5‐diazynylidene)‐4‐oxo‐3,4‐dihydronaphthalene‐1‐sulfonate
Cas Number:
11562-47-8
Molecular formula:
not applicable as UVCB
IUPAC Name:
3‐benzoyl‐2,6‐bis({[3‐(‐lambda5‐diazynylidene)‐4‐oxo‐3,4‐dihydronaphthalen‐1‐yl]sulfonyl}oxy)phenyl 3‐(‐lambda5‐diazynylidene)‐4‐oxo‐3,4‐dihydronaphthalene‐1‐sulfonate
Test material form:
solid

Test animals

Species:
mouse
Strain:
other: SHOE:NMRI
Details on species / strain selection:
The mouse is the recommended test species for this assay.
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Tierzucht Schönwalde GmbH i.G., Schönwalde, Germany
- Age at study initiation: appr. 8 weeks
- Weight at study initiation: males mean: 36.8 g, females mean: 29.8 g
- Housing: five animals per cage
- Diet: ad libitum rat/mice diet ssniff (ssniff GmbH, Soest, Germany)
- Water: tap water ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/- 3
- Humidity (%): 50+/-20
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
- Vehicle/solvent used: sesame oil

Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
On the day of the experiment, the test item was suspended in sesame oil. A magnetic stirrer was used to keep the preparation homogenous until administration.
Duration of treatment / exposure:
one treatment
Frequency of treatment:
one oral gavage treatment
Post exposure period:
Animals were killed 12, 24 or 48 h after dosing.
Doses / concentrationsopen allclose all
Dose / conc.:
2 000 mg/kg bw/day (nominal)
Remarks:
killing times: 12, 24 or 48 h, 5 males and 5 females each
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
killing times: 12, 24 or 48 h, 5 males and 5 females each
No. of animals per sex per dose:
5 per killing time
Control animals:
yes
yes, concurrent vehicle
Positive control(s):
cyclophosphamide
- Route of administration: oral (gavage)
- Doses / concentrations: 50 mg/kg bw

Examinations

Tissues and cell types examined:
erythrocytes
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION:
prelinimary toxicity study

DETAILS OF SLIDE PREPARATION:
A suspension of bone marow (taken from the femorae) and fetal bovine serum was formed and centrifuged for 5 minutes at app. 1200 rpm. The supernatant was discarded. The sediment was smeared on a clean slide. Staining was performed with Giemsa solution.

METHOD OF ANALYSIS:
1000 polychromatic erythrocytes were counted for each animal. The number of cells with micronuclei was recorded.
Evaluation criteria:
A substance is considered positive if there is a significant increase in the numbe rof micronucleated polychromatic erythrocytes for at least one of the time points compared with the concurent negative control group.
Statistics:
Wilcoxon-test (one-sided)

Results and discussion

Test results
Key result
Sex:
male/female
Genotoxicity:
negative
Toxicity:
no effects
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
No mortality and no clinical signs of toxicity were observed. No macroscopic findings were recorded at dissection.

Any other information on results incl. tables

Please refer to the attached background material.

Applicant's summary and conclusion

Conclusions:
The test item was not clastogenic in vivo under the conditions of the present study.
Executive summary:

The test substance was tested in an in vivo micronucleus test according to OECD Guideline 474. The test item was suspended in sesame oil and administered once via gavage at doses of 0 and 2000 mg/kg bw to male and female NMRI mice. The animals were killed either after 12, 24 or 48 h (5/sex/killing time). Endoxan (cyclophosphamide) was used as positive control at a dose of 50 mg/kg bw. The number of polychromatic and normochromatic erythrocytes containing micronuclei was not increased. The ratio of polychromatic/normochromatic erythrocytes in both male and female animals remained unaffected by the treatment with the test item and was statistically not different from the control values. The positive control induced a marked statistically significant increase in the number of polychromatic cells with micronuclei. The ratio of polychromatic erythrocytes to normocytes was not changes to a significant extent. Under the conditions of the study, th test item is not mutagenic in this test.

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