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EC number: 255-449-7 | CAS number: 41583-09-9
Genetic toxicity: in vivo
Administrative data
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
Data source
Referenceopen allclose all
- Reference Type:
- secondary source
- Title:
- Unnamed
- Year:
- 1 993
- Reference Type:
- publication
- Title:
- Evaluation of a Three-Exposure Mouse Bone Marrow Micronucleus Protocol: Results With 49 Chemicals
- Author:
- Shelby M.D. et al.
- Year:
- 1 993
- Bibliographic source:
- Environ. Mol. Mutagen., 21, 160-179
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- Deviations:
- yes
- Remarks:
- only 5-7 males per dose used, differing staining agent
- Principles of method if other than guideline:
- Standard protocol of the US National Toxicology Program
- GLP compliance:
- not specified
- Type of assay:
- other: micronucleus assay
Test material
- Reference substance name:
- Melamine
- EC Number:
- 203-615-4
- EC Name:
- Melamine
- Cas Number:
- 108-78-1
- Molecular formula:
- C3H6N6
- IUPAC Name:
- 1,3,5-triazine-2,4,6-triamine
Constituent 1
Test animals
- Species:
- mouse
- Strain:
- B6C3F1
- Sex:
- male
Administration / exposure
- Route of administration:
- intraperitoneal
- Vehicle:
- - Vehicle(s)/solvent(s) used: corn oil
- Details on exposure:
- Injection volume was 0.4 mL
- Duration of treatment / exposure:
- 3 days
- Frequency of treatment:
- one injection/day on 3 consecutive days
- Post exposure period:
- 1st study: For evaluation of bone marrow smears animals were sacrificed 24 hours after the 3rd exposure; for evaluation of peripheral blood smears
animals were sacrificed 48 hours after the 3rd exposure
2nd study: animals were sacrificed 24 hours after the 3rd exposure for both evaluation of bone marrow smears and evaluation of peripheral blood smears.
Doses / concentrationsopen allclose all
- Dose / conc.:
- 0 other: mg/kg bw
- Remarks:
- 1st study
- Dose / conc.:
- 500 other: mg/kg bw
- Remarks:
- 1st study
- Dose / conc.:
- 1 000 other: mg/kg bw
- Remarks:
- 1st study
- Dose / conc.:
- 2 000 other: mg/kg bw
- Remarks:
- 1st study
- Dose / conc.:
- 1 000 other: mg/kg bw
- Remarks:
- 2nd study
- Dose / conc.:
- 2 000 other: mg/kg bw
- Remarks:
- 2nd study
- No. of animals per sex per dose:
- 5 (exemption: only 3 animals were scored at the concentration of 2000 mg/kg bw in the first study)
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- 7,12-dimethylbenzanthracene (12.5 mg/kg)
Examinations
- Tissues and cell types examined:
- bone marrow and peripheral blood
- Details of tissue and slide preparation:
- DETAILS OF SLIDE PREPARATION:
Staining with acridine orange
METHOD OF ANALYSIS:
Bone marrow:
Slides were evaluated for the number of MN-PCE among 2000 PCE and for the percentage of PCE among 200 erythrocytes.
Peripheral blood:
According to the NTP-Standard protocol 2000 PCEs are evaluated for the number of MN-PCE and 1000 erythrocytes are evaluated for teh percentage of PCE among erythrocytes. - Evaluation criteria:
- A positive response is determined by statistical difference from the control group.
- Statistics:
- The data were analyzed using the Micronucleus Assay Data Management and Statistical software package (version 1.4 ), which was designed specifically for in vivo micronucleus data. The level of significance was set at an alpha level of 0.05.
To determine whether a specific treatment result ed in a significant increase in MN -PCE, the number of MN-PCE were pooled within each dose group and analyzed by a one- tailed trend test. In the software package used , the trend test incorporates a variance inflation factor to account for excess animal variability. In the event that the increase in the dose response cu rve is nonmonotonic , the software program allows for the data to be analyzed for a significant positive trend after data at the highest dose only has been excluded . However, in this event, the alpha level is adjusted to 0 .01 to protect against false positives.
The %PCE data were analyzed by an analysis of variance (ANOVA) test based on pooled data. Pairwise comparisons between each group and the concurrent solvent control group was by an unadjusted one-tailed Pearson chi -squared test which incorporated the calculated variance inflation factor for the study
Results and discussion
Test results
- Key result
- Sex:
- male
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Vehicle controls validity:
- valid
- Negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- No signs of toxicity were observed
Any other information on results incl. tables
Table 1: MN data analysis
Dose (mg/kg) |
MN-PCE/1000 (No. of animals scored) |
P value (a) |
Survival |
%PCEs (b) |
|
1st study, bone marrow (Sample collection time: 24 hours) |
0.003 |
||||
0 |
2.10 +/- 0.62 (5) |
5/5 |
57.3 |
||
500 |
1.80 +/-0.34 (5) |
0.685 |
5/5 |
56.6 |
|
1000 |
2.20 +/- 0.25 (5) |
0.439 |
5/5 |
57.1 |
|
2000 |
3.80 +/- 0.70 (5) |
0.013 |
6/6 |
56.1 |
|
Positive control |
6.90 +/- 0.87 (5) |
0.0001 |
- |
- |
|
2nd study, bone marrow (Sample collection time: 24 hours) |
0.340 |
||||
0 |
4.60 +/- 0.64 (5) |
5/5 |
60.9 |
||
1000 |
4.20 +/- 1.21 (5) |
0.596 |
5/5 |
51.7 |
|
2000 |
5.30 +/- 1.59 (5) |
0.344 |
5/6 |
56.3 |
|
Positive control |
8.40 +/- 0.56 (5) |
- |
- |
||
1st study, peripheral blood (c) (Sample collection time: 48 hours) |
0.964 |
||||
0 |
2.70 +/- 0.26 (5) |
5/5 |
1.2 |
||
500 |
3.50 +/- 0.91 (5) |
0.154 |
5/5 |
1.5 |
|
1000 |
2.40 +/- 0.56 (5) |
0.663 |
5/5 |
1.1 |
|
2000 |
1.50 +/- 0.76 (3) |
0.940 |
3/5 |
1.2 |
|
2nd study, peripheral blood (Sample collection time: 24 hours) |
0.411 |
||||
0 |
4.10 +/- 0.73 (5) |
no data available |
no data available |
||
1000 |
3.60 +/- 0.93 (5) |
0.716 |
no data available |
no data available |
|
2000 |
4.30 +/- 0.56 (5) |
0.413 |
no data available |
no data available |
|
Positive control |
8.40 +/- 0.40 (5) |
0.0001 |
no data available |
no data available |
(a) omitting the high dose value
(b) ((No. of PCEs)/(No. of PCEs + No. of NCEs)) x 100
(c) no data on positive control available
Applicant's summary and conclusion
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