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Diss Factsheets

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:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable deviations (bone marrow samples were collected only once after the last treatment. No informations about staining technique)
Cross-reference
Reason / purpose for cross-reference:
reference to same study

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Unnamed
Year:
2001
Reference Type:
secondary source
Title:
4-Nitrotoluene - CAS N°: 99-99-0 - SIDS Initial Assessment Report.
Author:
OECD
Year:
2003
Bibliographic source:
UNEP Publications

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Deviations:
yes
Remarks:
bone marrow samples were collected only once after the last treatment. No informations about staining technique
GLP compliance:
yes
Type of assay:
micronucleus assay

Test material

Constituent 1
Chemical structure
Reference substance name:
4-nitrotoluene
EC Number:
202-808-0
EC Name:
4-nitrotoluene
Cas Number:
99-99-0
Molecular formula:
C7H7NO2
IUPAC Name:
1-methyl-4-nitrobenzene
Details on test material:
- Name of test material (as cited in study report): p-nitrotoluene
- Analytical purity: 99%

Test animals

Species:
rat
Strain:
Fischer 344
Sex:
male

Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
- Vehicle(s)/solvent(s) corn oil
Details on exposure:
Rats were injected intraperitoneally
Duration of treatment / exposure:
72 hours
Frequency of treatment:
3 times at 24 hour interval
Post exposure period:
no
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 150, 300, 600 mg/kg bw in corn oil
Basis:
nominal conc.
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Positive control(s):
cyclophosphamide

Examinations

Tissues and cell types examined:
Rat bone marrow
Details of tissue and slide preparation:
The animals were killed 24 hours after the third injection, and blood smears were prepared from bone marrow cells
obtained from the femurs. Air-dried smears were fixed and stained; 2,000 polychromatic erythrocytes (PCEs) were
scored in up to five animals per dose group.
Evaluation criteria:
In the micronucleus test, an individual trial is considered positive if the trend test P value is less than or equal to 0.025 or if the P value for any single dosed group is less than or equal to 0.025 divided by the number of dosed groups. A final call of positive for micronucleus induction is preferably based on reproducibly positive trials. Ultimately, the final call is determined by the scientific staff after considering the results of statistical analyses, the reproducibility of any effects observed, and the magnitudes of those effects.
Statistics:
Statistical software package that tested for increasing trend over dose groups with a one-tailed Cochran-armitage trend test followed by pairwisecomparison between each dosed group and the control group (Integrated Laboratory Systems (ILS) (1990).Micronucleus Data Management and Statistical AnalysisSoftware, Version 1.4. ILS, P.O. Box 13501, Research Triangle Park, NC 27707.

Results and discussion

Test results
Sex:
male
Genotoxicity:
negative
Toxicity:
no effects
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
no signs of intoxication were reported; no increases in micronucleated PCE's in the bone marrow of male rats (P<=0.466):

Any other information on results incl. tables

Induction of micronuclei in bone marrow polychromatic erythrocytes of male rats treated with p-nitrotoluene by intraperitoneal injectiona

 Compound  Dose (mg/kg)  Number of rats with erythrocytes scores Micronucleated PCEs/1000 PCEb   P value c
 Corn oild  0  5  0.80 ± 0.12  
 p-nitrotoluene  150  5  1.00 ± 0.22  0.3186
   300  0.80 ± 0.12  0.5000
   600  5  0.90 ± 0.33  0l4041
       p=0.466 e  
 Cyclphosphamide f  25  5  10.30  ± 2.79  0.0000

a Study was performed at Integrated Laboratory Systems, Inc.

b PCE=polychromatic erythrocyte b Mean ± standard error

c Pairwise comparison with the vehicle control. Dosed group values are significant at P<=0.008; positive control values are significant at P<= 0.05

d Vehicle control

e Significance of micronucleated PCEs/1,000 PCEs tested by the one-tailed trend test; significant at P <=0.025

f Positive control

Applicant's summary and conclusion

Executive summary:

4-Nitrotoluene caused no increases in micronucleated polychromatic erythrocytes (PCEs) in the bone marrow of male rats given 0, 150, 300, or 600 mg/kg bw by intraperitoneal injection three times at 24 hour intervals. In male mice, treated with 0, 150, 300, or 600 mg/kg bw by intraperitoneal injection three times at 24 hour intervals, results of a first trial were considered positive, based on the responses of the two lowest doses; the trend test was not significant due to a downturn at the highest dose level. A second trial failed to induce a significant increase in micronucleated PCEs over the same dose range. Therefore the authors concluded the overall results as negative. Neither in the study with rats nor in the studies with mice were any signs of toxicity reported (U.S. Department of Health and Human Services, 2002).