Registration Dossier

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study run to a method comparable with current guidelines and to GLP

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Deviations:
no
GLP compliance:
yes
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
- Name of test material : 3-NITRO-1 ,2,4-TRIAZOL-5-0NE (NTO)
- Substance type: energetic explosive
- Physical state: light green to white crystalline solid with no odor
- Purity ca.99%
Specific details on test material used for the study:
Supplied by Ordnance Systems, Inc., Kingsport, Tennessee
Lot no.:BAE 07B 305001
Purity:99.6%

Test animals

Species:
rat
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
None stated

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
- Vehicle(s)/solvent(s) used: neat polyethylene glycol (PEG 200)

No additional data
Details on exposure:
None stated
Duration of treatment / exposure:
14 day
Frequency of treatment:
Daily
Post exposure period:
None stated
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 1000, 1500, and 2000 mg/kg
Basis:
actual ingested
No. of animals per sex per dose:
6 rats/sex/dose
Control animals:
yes, concurrent vehicle
Positive control(s):
The positive control group received a single dose of 200 mg/kg of EMS two days prior to study termination.

Examinations

Tissues and cell types examined:
The samples for micronucleus assay evaluations were prepared using the MicroFlow Plus Kit® (Litron Laboratories) following the manufacturer’s instructions. Briefly, peripheral blood (approximately 120 μL) was collected from the saphenous vein by puncturing the vein with an 18 gauge needle and collecting the blood using a pipette with a tip that was pre-coated in anti-coagulant.
Peripheral blood samples were processed for flow cytometric evaluation of micronucleated reticulocytes (MN-RET).
Details of tissue and slide preparation:
None stated
Evaluation criteria:
None stated
Statistics:
A one-way analysis of variance (ANOVA) was used to test for significant differences in %MN-RET for female and male rats separately. The Tukey multiple comparison test was used to evaluate the differences between dose groups. The results were considered to be statistically significant at p < 0.05. SPSS® version 16.0 was used for all analyses.

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
not specified
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
The frequency of micronucleated reticulocytes (%MN-RET) ranged from 0.20 to 0.23% in female rats and 0.21 to 0.27% in male rats treated with 1, 1.5, and 2 g/kg per day of the test substance in neat PEG 200. Treatment with the test substance did not statistically significantly (p = 0.096 and p = 0.616, respectively) increase the frequency of micronucleated reticulocytes (%MN-RET) in the peripheral blood of female or male rats. The positive control significantly increased, relative to the untreated control, the frequency of micronucleated reticulocytes in the peripheral blood of female and male rats (p = 0.005 and p = 0.047, respectively). These results indicate that the test substance is not genotoxic in rat peripheral blood under the test conditions.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
The test substance is not genotoxic in rat peripheral blood under the test conditions.