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Toxicological information

Genetic toxicity: in vivo

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Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP status unknown, near guideline study, published in peer reviewed literature, minor restrictions in design and/or reporting but otherwise adequate for assessment

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
1998

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Deviations:
yes
Remarks:
positive control substance tested only in males (considered not to affect scientific validity)
GLP compliance:
not specified
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
Obtained from Fluka

Test animals

Species:
mouse
Strain:
Balb/c
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 9 weeks
- Weight at study initiation: Approximately 23 g

ENVIRONMENTAL CONDITIONS - not reported

IN-LIFE DATES: Not reported

Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
- Vehicle(s)/solvent(s) used: mineral oil
Duration of treatment / exposure:
Administration by i.p. injection (two equal parts of dose given after a 24 hour interval). 2-stage model followed: First: 1 mL mineral oil or test substance at a dose equal to 80% of LD50 was dosed to males. Second: Doses administered equal to 40 and 80% of LD50 in males and 80% of LD50 in females.
Frequency of treatment:
Two i.p. injections, 24 hrs apart
Post exposure period:
30, 48 and 72 hours after the first injection bone marrow samples were collected
Doses / concentrations
Remarks:
Doses / Concentrations:
1800, 2960, 3600
Basis:
nominal conc.
No. of animals per sex per dose:
8 male and 4 female/0 mg/kg (solvent control - mineral oil); 12 male/1800 mg/kg; 12 female/2960 mg/kg; 24 male/3600 mg/kg
Control animals:
yes, concurrent vehicle
Positive control(s):
mitomycin C;
- Route of administration: not reported
- Doses / concentrations: 2.5 mg/kg; 0 mg/kg (solvent control - distilled water)

Examinations

Tissues and cell types examined:
Bone marrow polychromatic erythrocytes (NCEs)
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION: not reported
DETAILS OF SLIDE PREPARATION: bone marrow samples were collected from each mouse 30, 48 and 72 hours after 1st injection and 4 bone marrow smears were prepared from each mouse.

METHOD OF ANALYSIS: 1000 polychromatic erythrocytes (PCEs) per mouse were analysed for the number of micronucleated cells (mPCEs). The ratio of PCEs to mPCEs was determined by counting both cell types until the level of 200 NCEs was reached.
Evaluation criteria:
A test substance was considered genotoxic if the test substance induced a statistically significant and reproducible increase in the number of mPCEs for at least one of the study points.
Statistics:
One way analysis of variance with multiple comparison test by Domański, 1979

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
yes
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): No induction at 30, 48 or 72 hours after dosing
- Ratio of PCE/NCE (for Micronucleus assay): In males, the highest dose (80% of LD50) induced a statistically significant decrease in the PCEs to NCEs
ratio (cytotoxicity index 0.40 and 0.42 for 30 and 72 hours respectively, compared with 0.61 in control males. In the study on females using the equivalent of 80% of LD50, the animals survived but these doses were found not to induce cytotoxicity on bone marrow cells.

Any other information on results incl. tables

Induction of micronuclei in PCEs in mice

(Table based on Janik-Spiechowicz et al 1998, Mutation Research 412 (1998) 299-305, Table 2)

Dose

Number and sex

% mPCEs

ratio PCEs : mPCEs

harvest time (hours)

harvest time (hours)

30

48

72

30

48

72

Mesitylene

1 mL mineral oil

8 male

 

0.21 ± 0.08

 

 

0.61

 

1 mL mineral oil

4 female

 

0.20 ± 0.08

 

 

0.60

 

1800 mg/kg

12 male

0.20 ± 0.00

0.10 ± 0.09

0.17 ± 0.09

0.62

0.56

0.58

2960 mg/kg

12 female

0.17 ± 0.09

0.20 ± 0.00

0.22 ± 0.05

0.51

0.60

0.58

3600 mg/kg

24 male

0.24 ± 0.11

0.17 ± 0.05

0.14 ± 0.05

0.40*

0.33

0.42*

Mitomycin C (positive control)

1 mL distilled water

4 male

 

 

0.15 ± 0.05

 

 

1.37

2.5 mg/kg

12 male

4.15 ± 0.38*

4.22 ± 0.35*

1.25 ± 0.15*

0.71*

0.48*

0.14*

* p ≥ 0.05 data for males treated at 80% LD50 and mineral oil expressed as mean values ± S.D. for 2-stage experiments

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
Negative micronucleus response in polychromatic erythrocytes of the bone marrow.
Executive summary:

Mesitylene was tested in the in vivo micronucleus assay using bone marrow cells of the Imp:Balb/c mouse. There was no effect on the frequency of micronucleated polychromatic erythrocytes.