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Description of key information

LD50 was considered to be 1650 mg/kg (1380-1800) when SPF Wistar female rats were treated with 4-nitrobenzene-1,3-diamine orally by gavage.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Justification for type of information:
Data is from study report
Qualifier:
according to
Guideline:
other: as below
Principles of method if other than guideline:
Studies on the acute oral toxicity of 4-nitro-1,3-phenylenediamine in female Wistar rats
GLP compliance:
not specified
Test type:
standard acute method
Limit test:
no
Specific details on test material used for the study:
- Name of test material (as cited in study report): 4-nitro-m-phenylenediamine
- Molecular formula (if other than submission substance): C6H7N3O2
- Molecular weight (if other than submission substance): 153.14 g/mole
- Substance type: Organic
- Physical state: Solid
Species:
rat
Strain:
Wistar
Remarks:
SPF
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: SPF rats (Wistar strain TNO W 74, Ztichter: Winkelmann, Borchen)
- Age at study initiation: approximately 14 weeks old
- Weight at study initiation: average initial weight of 171 g.
- Fasting period before study: Not reported
- Housing: Conventional cages in Makrolon type III in a dust-free wood pellets in groups of 5 animals
- Diet (e.g. ad libitum): altromin R 1324, ad libitum
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: Not reported
- Identification: The identification of the animals was performed by picric acid marker of the skin and cage labeling

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±1.5 ° C
- Humidity (%): 60 ± 5 %
- Air changes (per hr): Not reported
- Photoperiod (hrs dark / hrs light): 12 hour light / dark cycle (artificial lighting 7-19 hours)

IN-LIFE DATES: From: To: Not reported
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
The substance was formulated at room temperature in water and administered by means of a rigid metal gavage at constant volume administered (20 ml / kg body weight) to 10 female animals/dose administered once.
Doses:
1000, 1500, 2000, 2500, 3100 mg/kg bw
No. of animals per sex per dose:
Totel: 50
1000 mg/kg bw: 10 female
1500 mg/kg bw: 10 female
2000 mg/kg bw: 10 female
2500 mg/kg bw: 10 female
3100 mg/kg bw 10 female
Control animals:
not specified
Details on study design:
On the day of application and in the following 14-day observation period, the animals were repeatedly inspected twice a day (once on weekends and holidays), for the nature, onset, duration and intensity of the clinical symptoms and possibly death.
Deaths and conclusion of the test animals slaughtered were randomly dissected
During the application and at the end of the 14 day observation period, the animals were weighed individually about living.
Statistics:
The calculation of the LD50 with the confidence interval for p <0.05 was carried out with the programmed probit analysis by Fink and Hund
Preliminary study:
not specified
Sex:
female
Dose descriptor:
LD50
Effect level:
1 650 mg/kg bw
Based on:
test mat.
95% CL:
1 380 - 1 800
Remarks on result:
other: 50 % mortality observed
Mortality:
when treated with 3100 mg/kg bw, all animals died
when treated with 2500 mg/kg bw, 9 female rats were died.
when treated with 2000 mg/kg bw, 7 female rats were died.
when treated with 1500 mg/kg bw, 5 female rats were died.
when treated with 1000 mg/kg bw, No mortality were observed.
Clinical signs:
Anesthesia, tremors, shaggy coat, stomach side position, reduction of the general condition, sedation were observed in treated female rats.
Body weight:
weight loss were observed in treated female rats.
Gross pathology:
No macroscopically specific findings were observed in treated rats died during the experiment and some at the end of the experiment killed animals.
Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
LD50 was considered to be 1650 mg/kg (1380-1800) when SPF Wistar female rats were treated with 4-nitrobenzene-1,3-diamine orally by gavage.
Executive summary:

In a acute oral toxicity study, SPF Wistar female rats were treated with 4-nitrobenzene-1,3-diamine in the concentration of 1000, 1500, 2000, 2500, 3100 mg/kg bw orally by gavage. All animals died at 3100 mg/kg bw, 9 female rats were died at 2500 mg/kg bw, 7 female rats were died at 2000 mg/kg bw, 5 female rats were died at 1500 mg/kg bw and No mortality were observed at 1000 mg/kg bw. In addition, Anesthesia, tremors, shaggy coat, stomach side position, reduction of the general condition, sedation and weight loss were observed in treated female rats. No macroscopically specific gross findings were observed treated female rats. Therefore, LD50 was considered to be 1650 mg/kg (1380-1800) when SPF Wistar female rats were treated with 4-nitrobenzene-1,3-diamine orally by gavage.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 650 mg/kg bw
Quality of whole database:
Data is Klimiach 1 and from study report

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Acute oral toxicity:

In different studies, 4-nitrobenzene-1,3-diamine has been investigated for acute oral toxicity to a greater or lesser extent. Often are the studies based on in vivo experiments data in rodents, i.e. most commonly in mice and rats for 4-nitrobenzene-1,3-diamine along with the study available on structurally similar read across substance 2-nitrobenzene-1,4-diamine (CAS no 5307-14-2).

In a experimental study conducted by Bayer AG Institute for Toxicology (L1384, T 4002138, 1982), SPF Wistar female rats were treated with 4-nitrobenzene-1,3-diamine in the concentration of 1000, 1500, 2000, 2500, 3100 mg/kg bw orally by gavage. All animals died at 3100 mg/kg bw, 9 female rats were died at 2500 mg/kg bw, 7 female rats were died at 2000 mg/kg bw, 5 female rats were died at 1500 mg/kg bw and No mortality were observed at 1000 mg/kg bw. In addition, Anesthesia, tremors, shaggy coat, stomach side position, reduction of the general condition, sedation and weight loss were observed in treated female rats. No macroscopically specific gross findings were observed treated female rats. Therefore, LD50 was considered to be 1650 mg/kg (1380-1800) when SPF Wistar female rats were treated with 4-nitrobenzene-1,3-diamine orally by gavage.

In another experimental study summarized by Cosmetic Ingredient Review (Journal of the American College of Toxicology, Volume 11, Number 4, 1992) by In a acute oral toxicity study, mice were treated with 4-nitrobenzene-1,3-diamine orally. 50 % mortality observed at 500 mg/kg bw in treated mice. Therefore, LD50 was considered to be 500 mg/kg bw when mice were treated with 4-nitrobenzene-1,3-diamine orally.  

Also it is further supported experimental study conducted by Bayer AG Institute for Toxicology (L1384, T 4002138, 1982), SPF Wistar male rats were treated with 4-nitrobenzene-1,3-diamine in the concentration of 1000, 1500, 2000, 2500, 3100 mg/kg bw orally by gavage. All animals died at 3100 mg/kg bw, 8 male rats were died at 2500 mg/kg bw, 4 male rats were died 2000 mg/kg bw, 1 male rats were died at 1500 mg/kg bw and No mortality were observed at 1000 mg/kg bw. Sedation, anesthesia, abdominal / side position, tremor, rough coat, reduction of the general condition were observed in treated male rats. No macroscopically specific findings were observed in treated rats died during the experiment and some at the end of the experiment killed animals. Therefore, LD50 was considered to be 2050 mg/kg (1800- 2300) when SPF Wistar male rats were treated with 4-nitrobenzene-1,3-diamine orally by gavage.

 

 

Further it is supported by experimental study conducted by Lloyoet al(Food Cosmetics Toxicology. Vol. 15, pp. 607-610. 1977) on structurally similar read across substance 2-nitrobenzene-1,4-diamine (CAS no 5307-14-2), CFY male and female rats were treated with 2-nitrobenzene-1,4-diamine in the concentration of 1800 mg/kg (40 %) as solutions or suspensions in 0.5% aqueous gum tragacanth, containing 0.05% Na2SO3 orally by gavage. 50% mortality was observed in treated male and female rats at 1800 mg/kg bw. Lethargy and piloerection were observed in treated rat. Other reactions elicited by more than one of the test compounds included increased salivation, ataxia, fine body tremors, changes in respiratory rate, dieresis and diarrhea were observed in treated rat. Autopsy of the animals that died as a result of treatment revealed changes included darkening of the liver and kidneys, darkening or pallor of the spleen, haemorrhage of the lungs and intestines, and injection of the intestinal and mesenteric blood vessels. Autopsy of the survivor rats at the end of each experiment did not reveal any abnormalities indicative of residual systemic effects. Therefore, LD50 was considered to be 1800 mg/kg bw(1500-2300) when CFY male and female rats were treated with 2-nitrobenzene-1,4-diamine orally.  

Thought, the LD50 2050 mg/kg value for male rat is available, since the female are more sensitive than male, the LD50 for 4-nitrobenzene-1, 3-diamine is considered to be 1650 mg/kg.

Thus, based on the above studies on 4-nitrobenzene-1, 3-diamine and its read across substances, it can be concluded that LD50 value is less than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, 4-nitrobenzene-1,3-diamine can be classified as “Category IV” of acute oral toxicity.

Justification for classification or non-classification

Based on the above predictions and studies on 4-nitrobenzene-1, 3-diamine and its read across substances, it can be concluded that LD50 value is less than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, 4-nitrobenzene-1,3-diamine can be classified as “Category IV” of acute oral toxicity.