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EC number: 246-836-1 | CAS number: 25321-14-6
- 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
Developmental toxicity / teratogenicity
Administrative data
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 20 Feb 1980 - 15 Sep 1980
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Reliability as cited in OECD SIDS Dinitrotoluene (isomers mixture) CAS No.: 25321-14-6, 2004. Comparable to guideline study with acceptable restrictions (insufficient number of DNT-treated pregnant animals per dose (6-13))
Cross-reference
- Reason / purpose for cross-reference:
- reference to same study
Data source
Referenceopen allclose all
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 982
- Report date:
- 1982
- Reference Type:
- publication
- Title:
- Teratological and postnatal evaluation of dinitrotoluene in Fischer 344rats
- Author:
- Price, C.J. et al.
- Year:
- 1 985
- Bibliographic source:
- Fundamental and Applied Toxicolgy 5, 948-961
- Reference Type:
- publication
- Title:
- Teratological evaluation of technical grade dinitrotoluene in Fischer 344rat
- Author:
- Wolkowski-Tyl, R. et al.
- Year:
- 1 981
- Bibliographic source:
- Teratology 23, 70A
- Reference Type:
- secondary source
- Title:
- Dinitrotoluene (isomers mixture), CAS No.: 25321-14-6
- Author:
- OECD SIDS
- Year:
- 2 004
- Bibliographic source:
- OECD SIDS
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- Deviations:
- yes
- Remarks:
- lower number of DNT-treated pregnant animals per dose (6-13 instead of 20)
- GLP compliance:
- not specified
- Limit test:
- no
Test material
- Reference substance name:
- Dinitrotoluene
- EC Number:
- 246-836-1
- EC Name:
- Dinitrotoluene
- Cas Number:
- 25321-14-6
- Molecular formula:
- C7H6N2O4
- IUPAC Name:
- 1-methyl-2,4-dinitrobenzene; 2-methyl-1,3-dinitrobenzene
- Details on test material:
- - Name of test material (as cited in study report): technical grade dinitrotoluene (DNT)
- Analytical purity: 2,4-DNT (76%), 2,6-DNT (19%), 3,4-DNT (2.4%), 2,3-DNT (1.5%), 2,5-DNT (<1%) and 3,5-DNT (<1%) .
Constituent 1
Test animals
- Species:
- rat
- Strain:
- Fischer 344
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Breeding Laboratory, Inc ., Wilmington, MA, and Portage, MI
- Both males and females were approximately 56 days of age at the time of arrival at Research Triangle Institute (RTI), thus providing animals for the study which were young, but sexually mature at the time breeding was initiated. Males and females were bred between 3-7 months after arrival at RTI.
- Weight at study initiation: 145-226 g at gestational day 0 (females were matched across dose groups by body weight on gestational day 0)
- Housing: males were housed singly and females were group housed (4-per-cage)
- Diet and water: Purina Certified Rodent Chow (5002) and tap water were available ad libitum throughout the study
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.22 +/- 1.11
- Air changes (per hr): 12 to 14 times per hour
- Photoperiod (hrs dark / hrs light): 12/12
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- corn oil
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
- Dose volume: 2 mL/kg
- Dosage solutions of DNT were formulated at concentrations of 7.00, 17.50, 18.75, 37.50, 50.00 and 75.00 mg/mL.
- Dosing solutions were stored at 5°C, and used for dosing up to 8 days after formulation
VEHICLE
- purity: laboratory-grade
- Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- High performance liquid chromatography (HPLC) in the normal phase mode was used for the analysis of DNT dosage formulations.
Samples were quantified from the absolute peak height of the major DNT isomer ( i.e. 2,4-DNT) measured manually and results calculated from
an external standard calibration curve. - Details on mating procedure:
- - Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1:1
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: presence of sperm in vaginal smear referred to as day 0 of pregnancy - Duration of treatment / exposure:
- 14 days: gestational day 7 through 20
- Frequency of treatment:
- Daily
- Duration of test:
- Sacrifice on gestational day 20
Doses / concentrations
- Remarks:
- Doses / Concentrations:
0, 14, 35, 37.5, 75, 100, and 150 mg/kg/day
Basis:
actual ingested
- No. of animals per sex per dose:
- test concentration [mg/kg/day]: total no. of animals (pregnant and non-pregnant)
14: 22
35: 13
37.5: 22
75: 13
100: 23
150: 13
vehicle control: 37
positive control: 36 - Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Positive control: Hydroxyurea, 200 mg/kg bw/day in sterile distilled water, p.o.
- Dose selection rationale: is based upon pilot data which indicated that exposure of timed-pregnant Fischer 344 rats to DNT (75 or 150 mg/kg/day, po) on gd 9 through 12 was well tolerated
Examinations
- Maternal examinations:
- CLINICAL SIGNS OF TOXICITY:
- Time schedule: on gestational days (gd) 0, and 7 through 20 (prior to daily dosing)
BODY WEIGHT: Yes
- Time schedule for examinations: on gestational days (gd) 0, and 7 through 20 (prior to daily dosing)
FOOD CONSUMPTION
- Time schedule for examinations: determined twice a week, beginning on day 0 of gestation
- Calculation of average food intake (g) per animal per day
WATER CONSUMPTION
- Time schedule for examinations: determined twice a week, beginning on day 0 of gestation
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- liver weight, spleen weight
OTHER
- hematology on day of sacrifice: Methemoglobin assay, reticulocyte count, complete blood count - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early, middle, late, and full resorptions: Yes - Fetal examinations:
- - For each fetus and "full" resorption: body weight, crown-rump length, placental weight and sex
- External examinations: Yes; all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: all per litter
- Head examinations: Yes: all per litter
- Other: hematology: Methemoglobin assay, reticulocyte count, complete blood count - Statistics:
- Due to the practical considerations in processing large numbers of dams and fetuses, an experimental design with three replicates was employed. Data were pooled across replicates and nonparametric statistics were applied: Kruskal-Wallis one-way analysis of variance by ranks (Siegel, 1956) was used to test for differences among DNT-treated and vehicle control groups. Kruskal-Wallis was significant at p<0 .05, the Mann-Whitney U test was
utilized to make individual comparisons between vehicle and each DNT treated group (Siegel, 1956). Jonckbeere's test for k independent samples (Jonckheere, 1954) was employed to identify significant dose response trends. Nominal level measurements were analyzed by Fisher's Exact Test (pairwise comparisons) or Chi Square (comparisons among dose groups) (Siegel, 1956) . Statistical comparisons between vehicle control and positive control, ( i .e ., hydroxyurea-treated) groups were conducted using the Mann-Whitney U test (Siegel, 1956) or Fisher's Exact Test (Siegel, 1956). Statistical analysis of hematological data from methemoglobin analysis (RTI), differential and complete blood counts (CIIT) were performed using a t-test to compare vehicle control to hydroxyurea or DNT treatment groups. The alpha level for each statistical comparison was 0.05 and the significance level (two-tailed) for each statistical comparison was reported at p<0.05 or p<0.01 . - Historical control data:
- No data
Results and discussion
Results: maternal animals
Maternal developmental toxicity
- Details on maternal toxic effects:
- Maternal toxic effects:yes
Effect levels (maternal animals)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 14 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Basis for effect level:
- other: maternal toxicity
Maternal abnormalities
- Key result
- Abnormalities:
- effects observed, treatment-related
- Localisation:
- other: relative spleen weights; secondary effects on hematology were observed at higher doses
- Description (incidence and severity):
- relative spleen weights; secondary effects on hematology were observed at higher doses (See "Remarks on results including tables and figures").
Results (fetuses)
- Details on embryotoxic / teratogenic effects:
- Embryotoxic / teratogenic effects:no effects
Effect levels (fetuses)
open allclose all
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- >= 150 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: fetotoxicity
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- >= 150 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: embryotoxicity
Fetal abnormalities
open allclose all
Overall developmental toxicity
- Key result
- Developmental effects observed:
- no
- Lowest effective dose / conc.:
- 150 mg/kg bw/day (nominal)
- Treatment related:
- yes
Any other information on results incl. tables
MATERNAL TOXICITY
Clinical signs and maternal mortality
The clinical signs related to DNT treatment were rough coat, lethargy and hind limb weakness observed in 7/13 high-dose females.
Treatment |
Vehicle |
Hydroxyurea |
Dinitrotoluene |
||||||
Dose (mg/kg/d) |
- |
200 |
14 |
35 |
37.5 |
75 |
100 |
150 |
|
Females treated |
37 |
36 |
22 |
13 |
22 |
13 |
23 |
13 |
|
% Pregnant (gd 20) |
91 |
91 |
77 |
100 |
92 |
86 |
92 |
83 |
|
% Deaths (gd 0-20) |
0 |
0 |
4.5 |
7.7 |
0 |
0 |
4.3 |
46.2 |
Maternal weight gain
A significant treatment effect was observed for maternal weight gain during gestation, but only the highest dose group (150 mg/kg/day) showed a significant decrease in weight when pairwise comparisons were made with the vehicle control group. Absolute maternal weight gain (i.e., maternal weight gain during treatment minus gravid uterine weight) exhibited a significant dose-related decrease across DNT treatment groups; significant reduction of absolute maternal weight gain was observed at 14, 100 and 150 mg/kg/day DNT. Significant dose-related increases in relative maternal liver and spleen weights were observed across DNT treatment groups.
Water consumption did not appear to be affected by DNT treatment, but statistical analysis was not performed since average consumption was based on group housed animals which were not equally distributed across cages.
Maternal weight gain on gestational days 7 through 20
Treatment |
Vehicle |
Hydroxyurea |
Dinitrotoluene |
|||||
Dose (mg/kg/d) |
- |
200 |
14 |
35 |
37.5 |
75 |
100 |
150 |
Number sacrificed |
20 |
20 |
10 |
7 |
12 |
6 |
12 |
5 |
Total wt gain (g) |
61.8 ± 3.6* |
54.0 ± 4.6 |
64.9 ± 4.6 |
66.1 ± 6.0 |
55.8 ± 5.8 |
64.8 ± 6.8 |
52.7 ± 4.5 |
8.1 ± 20.1** |
Absolute wt. gain (g) |
24.1 ± 2.1 |
21.1 ± 1.7 |
17.2 ± 3.9* |
26.7 ± 4.3 |
22.2 ± 2.1 |
23.1 ± 2.1 |
15.2 ± 1.9** |
-14.0 ± 13.4** |
Liver wt (% body wt) |
4.09 ± 0.08** |
3,95 ± 0.05 |
3.91 ± 0.1* |
4.12 ± 0.09 |
3,96 ± 0.09 |
4.55 ± 0.1** |
4.58 ± 0.08** |
4.79 ± 0.36 |
Spleen wt (% body wt) | 0,197 +/- 0,003** | 0,209 +/- 0,009 | 0,185 +/- 0,007 | 0,223 +/- 0,011* | 0,215 +/- 0,006* | 0,246 +/- 0,010** | 0,320 +/- 0,027** | 0,284 +/- 0,059* |
*p<0.05
**p<0.01
Maternal hematological examination
No significant difference was observed in hematological parameters for dams sacrificed at 20 minutes or 120 minutes after final treatment on gd 20; therefore, data were pooled for analysis. At sacrifice on gd 20, dams treated with 100 mg/kg/day DNT exhibited statistically significant increases in methemoglobin, reticulocyte count, red blood cell size, red blood cell distribution width and platelet count. Statistically significant decreases in red blood cell count and hematocrit were also observed.
FETAL TOXICITY
- no significant effects on % resorptions, % dead or live fetuses;
- no significant effects on live litter size, fetal body weight, crown-rump length, sex distribution, fetal grouwth and morphological development;
- changes of fetal liver and spleen weights without dose dependence and also occurring in control groups
- 100 mg: changes of hematological parameters (as in dams), decreased reticulocyte count, decreased RBC count, and increased RBC size
- no statistically significant increased malformations at any dose: % malformed fetuses per litter (external, visceral, sceletal) in dose groups 14/35/37.5/75/100/150 mg/kg and vehicle/pos. control: 4.1/3.2/0.8/0.0/5.8/0.0 and 3.8/30.6
Total number of animals and pregnants evaluated in the study and total number of live litter and live fetuses examined:
mg/kg bw/day | total number of animals | total number of pregnants | number of live litters examined | number of live fetuses examined |
14 | 22 | 13 | 10 | 9,2 +/- 0,7 |
35 | 13 | 7 | 7 | 9,0 +/- 0,9 |
37.5 | 22 | 13 | 12 | 6,4 +/- 1,1 |
75 | 13 | 7 | 6 | 8,3 +/- 1,3 |
100 | 23 | 13 | 12 | 7,3 +/- 0,9 |
150 | 13 | 6 | 2 | 7,3 +/- 1,7 |
Vehicle control | 37 | 22 | 20 | 7,3 +/- 0,7 |
Positive control | 36 | 22 | 19 | 7,7 +/ 0,6 |
DOSAGE FORMULATIONS
Calculated dosages based upon analysis of DNT and hydroxyurea formulations are given below :
Nominal Treatment (mg/kg/day) | Theoretical Treatment Mean and Range (mg/kg/day ) |
Hydroxyurea (200) | 185.0 (175 .2-200 .4) |
DNT (14) | 12.1 [8 .5-17 .1] |
DNT (35) | 33.8 [28 .7-41 .2] |
DNT (37.5) | 38.4 [15 .5-70 .4] |
DNT (75) | 75.0 [71 .6-81 .9] |
DNT (100) | 110.5 [65 .6-155 .3] |
DNT (150) | 157.4 [125 .6-177 .3] |
Since dams received doses from as many as three individually formulated solutions during gestation and since a wide range of measured concentrations was represented within these formulations, it was not possible to confirm the actual oral exposure for individual dams. Discrepancies between dosage formulation records and the results of HPLC analyses may have been in part due to prolonged storage (6 to 11 months) prior to analysis.
Applicant's summary and conclusion
- Conclusions:
- DNT was not teratogenic in Fischer 344 rats even at dosages which produced significant dose-related signs of maternal and fetal toxicity .
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