[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

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2009-04-23 till 2009-07-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline-conform study under GLP without deviations.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.11 (Mutagenicity - In Vivo Mammalian Bone-Marrow Chromosome Aberration Test)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5385 (In Vivo Mammalian Cytogenetics Tests: Bone Marrow Chromosomal Analysis)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

chromosome aberration assay

Test material

Test material information

Constituent 1

Details on test material:

Batch No.: FEB 279-810

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Age at study initiation: minimum 7 weeks
- Weight at study initiation: the weight variation of animals should be minimal and not exceed ± 20% of the mean weight of each sex
- Assigned to test groups randomly: yes
- Housing: 5 animals of identical sex per cage
- Diet (e.g. ad libitum): Free access to Altromin 1324 maintenance diet
- Water (e.g. ad libitum): Free access to tap water, sulphur acidified to pH value of approximately 2.8 (drinking water, municipal residue control, micro -biologically controlled periodically)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 55 ± 10%
- Air changes (per hr): at least 10 x per hour
- Photoperiod (hrs dark / hrs light): 12 hours light, 12 hours dark

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

Cottonseed Oil

Duration of treatment / exposure:

The exposition times were 24 h for all dose groups evaluated and 48 h for the additional negative control and highest dose group

Frequency of treatment:

The animals received the test item once.

Post exposure period:

48 h

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
200 mg/kg bw
Basis:

Doses / concentrations 2

Remarks:

Doses / Concentrations:
500 mg/kg bw
Basis:

Doses / concentrations 3

Remarks:

Doses / Concentrations:
1000 mg/kg bw.
Basis:

No. of animals per sex per dose:

5

Control animals:

yes

Positive control(s):

Cyclophosphamide
- Route of administration: ip, single
- Doses / concentrations: 10 mL/kg bw

Examinations

Tissues and cell types examined:

bone marrow cells

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Remarks:

1000 mg/kg

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

RESULTS OF RANGE-FINDING STUDY:

One female and one male rat received a single dose of 2000 mg/kg bw ip and died within 26.5 hours for the female and 27.5 hours for the male after application of the test item. Three female and three male rats received a single dose of 1000 mg/kg bw ip and showed toxic symptoms as reduction of spontaneous activity, prone position, constricted abdomen and an abnorm pinna reflex, but survived 72 hours after the treatment. (Table 1). Due to the results obtained in the pre-experiment 1000 mg/kg was chosen as maximum tolerable dose (1 MTD) in the main experiment.

RESULTS OF DEFINITIVE STUDY:

For all dose groups, 100 metaphases per animal were scored for the incidence of structural chromosomal aberrations.

The negative controls (24 h and 48 h) evaluated were within the range of the historical control data (male: 0.0% - 5.1%, female: 0.0% - 3.7, Table 14). The mean values of aberrant cells observed for the negative control (24 h) were 0.6% for male and 0.2% for female rats. The mean values for the 48 h negative control were 0.4% for the male and 1.2% for the female rats.

The dose group treated with 1000 mg/kg bw (24 h treatment) showed mean values of 0.4% for male and 0.2% for female rats. The dose group treated with 500 mg/kg bw (24 h treatment) showed mean values of 0.6% for male and 1.2% for female rats. The dose group treated with 200 mg/kg bw (24 h treatment) showed mean values of 0.6% for both, the male and female rats. The mean values observed for the 1000 mg/kg bw (48 h treatment) were 0.6% as well for both, the male and female rats.

All mean values of aberrant cells noted after treatment with the test item were within the historical control data range of the negative control (Table 14). No biologically relevant increase of aberrant cells was found.

The nonparametric Mann-Whitney test was performed to verify the results. No statistically significant enhancement (p<0.05) of aberrant cells was noted at any dose group of the test item evaluated .

Cyclophosphamide (40 mg/kg bw) administered ip was used as positive control which induced a significant increase of induced aberrant cells frequency (percentage of aberrant cells was 54.4% for male and 44.8% for female rats). This demonstrates the validity of the assay.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative
The test item did not induce structural chromosomal damage in bone marrow cells of the rat.
Therefore, the test item is considered to be non-clastogenic in this Mammalian Bone Marrow Chromosome Aberration Test under the experimental conditions reported.