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EC number: 201-853-3 | CAS number: 88-72-2
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Genetic toxicity: in vivo
Administrative data
- Endpoint:
- in vivo mammalian cell study: DNA damage and/or repair
- Remarks:
- Type of genotoxicity: DNA damage and/or repair
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline with acceptable restriction (only 90 cells scored /dose, results of the positive control not given, postive control not guideline listed and no rational for choice is given)
Data source
Reference
- Reference Type:
- publication
- Title:
- Unnamed
- Year:
- 1 992
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 486 (Unscheduled DNA Synthesis (UDS) Test with Mammalian Liver Cells in vivo)
- Deviations:
- yes
- Remarks:
- total of 90 cells scored/ dose, positive control not listed in guideline
- Principles of method if other than guideline:
- According to the method of Mirsalis et al (1985) Carcinogenesis 6, 1521-1524
- GLP compliance:
- yes
- Type of assay:
- unscheduled DNA synthesis
Test material
- Reference substance name:
- 2-nitrotoluene
- EC Number:
- 201-853-3
- EC Name:
- 2-nitrotoluene
- Cas Number:
- 88-72-2
- Molecular formula:
- C7H7NO2
- IUPAC Name:
- 1-methyl-2-nitrobenzene
- Details on test material:
- - Source: Aldrich Chemical Co. (Milwaukee, WI, USA),
- Analytical purity: >96%
- Impurities: < 1% (mostly m- and p-nitrotoluene)
- Storage: RT
- Stability: reanalysis performed at approx. 4 months intervals indicated that the test substance was stable under the storage conditions chosen
Constituent 1
Test animals
- Species:
- rat
- Strain:
- Fischer 344
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- - Source: Taconic Farms, Inc. (Germantown, NY
- Age at study initiation: 11 to 12 weeks
- Housing: 3/cage
- Acclimation period: 7-8 weeks
OTHER: Assignment to treatment groups; by weight class using a computer-generated randomization procedure.
ENVIRONMENTAL CONDITIONS: not reported
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- Vehicle: Corn oil
- Details on exposure:
- DOSE VOLUME: 5 ml/kg bw
- Duration of treatment / exposure:
- 12, 24h
- Frequency of treatment:
- single application
- Post exposure period:
- no
Doses / concentrations
- Remarks:
- Doses / Concentrations:
0, 100, 200, or 500 mg/kg (male rats); 0, 200, 500, or 750 mg/kg (female rats)
Basis:
other: nominal
- No. of animals per sex per dose:
- 12h: 3
24h: 3 - Control animals:
- yes, concurrent vehicle
- Positive control(s):
- 2,6-dinitrotoluene
Examinations
- Tissues and cell types examined:
- Hepatocytes
- Details of tissue and slide preparation:
- CRITERIA FOR DOSE SELECTION: no data
TREATMENT AND SAMPLING TIMES (in addition to information in specific fields): 12h and 24h after gavage, 3 rats were selected from each group for the collection of hepatocytes for UDS determination. Animals were anesthetized with sodium pentobarbitol, and the livers were perfused with Hanks¿ balanced salts containing ethylene glycol-bis (baminoethylether)-N,N-tetra-acetic acid (EGTA) and HEPES buffer at pH 7.2 for 2 ¿ 4 minutes and with a collagenase solution for 4-10 minutes. The liver was removed and hepatocytes were obtained by mechanical dispersion of the excised liver tissue in Williams¿ Medium C with collagenase.
DETAILS OF SLIDE PREPARATION:
Cells were cultured on plastic coverslips in Williams Medium E for 1.5 to 2 hours at 35°C.
Unattached cells were then removed and cultures refed with 2.5 ml Williams¿ Medium E (without fetal bovine serum) containing 3H-thymidine for 4h. Cell cultures were found to contain >80% viable cells. The cultures were refed with Williams¿ Medium E (without fetal bovine serum) containing unlabeled thymidine, and returned to the incubator for 14 to 19 hours after which they were washed (Williams¿ Medium E without serum) and the nuclei swollen by the incubation with 1% sodium citrate (8-12min) and fixed in glacial acetic acid: ethanol (1:3), washed (deionized water) and dried for for at least 24h. The coverslips were mounted on glass slides, dipped in Kodak NTB2 photographic emulsion, and dried. Coated slides were stored for 7 days at 4°C in light-tight boxes containing Drierite. The emulsions were developed in D19, fixed, and stained with Williams¿ modified hematoxylin and eosin procedure.
METHOD OF ANALYSIS:
The cells were examined microscopically at approximately 1500X magnification under oil immersion. UDS was measured by counting nuclear grains and subtracting the largest number of grains from 3 nuclear-sized areas adjacent to each nucleus (background count).
This value is referred to as the net nuclear grain count (NNG) and can be a negative number if the number of grains in any background area exceeds the number of grains in the nucleus. The NNG was determined for 30 randomly selected cells on each coverslip. The mean NNG was determined from triplicate coverslips, if available (90 total nuclei), for each treated animal (2 or 3 animals per dose level). The percentage of cells in S-phase was calculated as those cells exhibiting nuclei blackened by grains too numerous to count.
Approximately 2000 cells were counted from randomly selected areas of each slide. For each dose, 3 slides were scored for each of 3 animals (6000 total cells). - Evaluation criteria:
- According to the srtudy report, the test was considered positive if an increase in the mean net nuclear grain count was observed to at least 5 grains per nucleus in excess of the concurrent vehicle control, or the percentage of nuclei with 5 or more net grains was increased above 10% of the examin d population, in excess of the concurrent vehicle control.
The test is considered negative if the mean net grain count for all groups is less than 1 above the concurrent control value, and/or the percent of nuclei with 5 or more net grain counts does not increase more than 2% above the concurrent vehicle control. - Statistics:
- Significance of the response was determined using the Student¿s t-test modified for unpaired observations with unequal variances.
Results and discussion
Test results
- Sex:
- male/female
- Genotoxicity:
- positive
- Remarks:
- Statistically significant increase of UDS from 100 mg/kg bw in male rats and from 200 mg/kg bw in female rats. Also an increase of the number of hepatocytes in S-phase, cultured from both male (at 500 mg/kg bw) and female (>= 200 mg/kg bw) rats was seen
- Toxicity:
- not specified
- Vehicle controls validity:
- valid
- Positive controls validity:
- not specified
- Additional information on results:
- Along o-nitrotoluene, m-, and p- nitrotoluene were also included in the panel of substances tested in the rat in vivo/in vitro UDS test. A comparison of UDS activity of the 3 nitrotoluene isomers was performed using identical doses (0, 100, 200, 500 mg/kg bw) given to male F344 rats. From these data, it was apparent that the o-nitrotoluene was the only isomer which was positive for induction of UDS.
o-Nitrotoluene was also found to increase the number of hepatocytes in S-phase, cultured from both male and female rats (see table 1). In this assay, neither m- or p-nitrotoluene caused an increase in S-phase hepatocytes in either sex of the rats (data not shown in documentation).
Any other information on results incl. tables
Table 1:Unscheduled DNA Synthesis in Male and Female Rats and Percent Liver Cells in S-Phase (12 and 24 Hours after Single Oral Gavage Dose of o-Nitrotoluene, respectively)
Dose |
Male |
Female |
||
mg/kg bw |
UDS (12h after gavage) |
Cells in S phase (24 h after gavage) |
UDS (12h after gavage) |
Cells in S phase (24 h after gavage |
mean net nuclear grain count ± SD |
% |
mean net nuclear grain count ± SD |
% |
|
0 |
-2.57 ± 0.18 |
0.66 ± 0.18 |
-5.96 ± 0.59 |
0.58 ± 0.27 |
100 |
- 0.05 ± 0.47* |
0.86 ± 0.42 |
Not tested |
Not tested |
200 |
5.64 ± 0.57** |
3.61 ± 0.94 |
- 2.24 ± 1.0** |
2.40 ± 0.70 |
500 |
13.11 ± 1.14** |
3.20 ± 0.47* |
0.94 ± 0.93** |
7.17 ± 0.70 |
750 |
Not tested |
Not tested |
1.45 ± 0.93* |
12.98 ± 3.9 |
* Significantly different from control group (P=0.05)
** Significantly different from control group (P=0.01)
Applicant's summary and conclusion
- Executive summary:
NTP, 1992
Groups of 6 male F344/ rats/dose and 6 female F344 rats/dose were administered oral doses of o-Nitrotoluene in corn oil via gavage; 0; 100; 200; 500 mg/kg for males and 0; 200; 500 and 750 mg/kg bw for females, respectively, according to a method similar to OECD guideline 486 (total of 90 cells scored/ dose, positive control not listed in guideline) Perfusion of liver and preparation of hepatocytes was performed 12h and 24h after gavage.
o-Nitrotoluene induced DNA damage in mammalian liver cells in vivo (male and female F344 rats) that could be repaired by unscheduled DNA synthesis in vitro; Male rats (>= 100 mg/kg bw), Female rats (>= 200 mg/kg bw).
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