4-amino-3-nitrophenol

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 1992-03-05 to 1992-05-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Analysis certificate not available during the study. Another study report and SCCS Opinion provided the certificate of analysis the same batch (CIT Study, 8796 TAL).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

24th February 1987

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

4-amino-3-nitrophenol

EC Number:

210-236-8

EC Name:

4-amino-3-nitrophenol

Cas Number:

610-81-1

Molecular formula:

C6H6N2O3

IUPAC Name:

4-amino-3-nitrophenol

Test material form:

solid: crystalline

Remarks:

Dark red crystals

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: op 238
- Expiration date of the lot/batch: no data
- Purity test date:no data

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature
- Stability under test conditions: not specified
- Solubility and stability of the test substance in the solvent/vehicle:not specified

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Final preparation of a solid: the test substance was ground down to a fine dust with a mortar and a pestle then suspended in the vehicle.
FORM AS APPLIED IN THE TEST (if different from that of starting material): 99% suspension in 1,2-propanediol, at a volume of 10 mL/Kg

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Iffa Crédo, 69210 L'Arbresle, France
- Females (if applicable) nulliparous and non-pregnant: not specified
- Age at study initiation: approximatively 6 weeks old
- Weight at study initiation: males : 173±10g, females : 139±5g
- Fasting period before study: 18 hours before administration
- Housing: in group of 5 animals of the same sex in sterilizable polycarbonate cages (48x27x20cm) covered with a stainless steel lid containing food and water bottle.
- Diet (e.g. ad libitum): certified pellet diet "Rats - Mice sustenance ref. A04 C" (U.A.R., France). An analysis of the quality of the food and the major contaminants (pesticides, heavy metals, mycotoxins etc.) was performed by the supplier and given for each batch.
- Water (e.g. ad libitum): Free access to tap water filtered by a 0.22 micron membrane (Société Millipore, France) and contained in water bottles. Bacteriological and chemical analyses of the water and the detection of the major contaminants (pesticides, heavy metals and nitrosamines) were made periodically (Laboratoire municipal de Rouen, France)
- Acclimation period: at least 5 days before instillation
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°C
- Humidity (%): 50±20% relative humidity
- Air changes (per hr): the air was non-recycled and filtered by absolute filters
- Photoperiod (hrs dark / hrs light): 12 hours of light/12 hours of dark
IN-LIFE DATES: From:1992-03-27 to 1992-04-15 and from 1992-04-18 to 1992-05-07

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 1,2-propanediol

Details on oral exposure:

VEHICLE
- Concentration in vehicle: adjusted to each rats bodyweight, test item suspended as 99% in vehicle
- Amount of vehicle (if gavage): 10ml/kg
- Justification for choice of vehicle: no data
- Lot/batch no. (if required): No. 1256840 (Aldrich Chimie, France)
- Purity: no data
MAXIMUM DOSE VOLUME APPLIED: 10mg/kg
DOSAGE PREPARATION (if unusual): the test substance was ground down to a fine dust with a mortar and a pestle then suspended in the vehicle.

Doses:

500/100/1500 mg/kg bodyweight

No. of animals per sex per dose:

5 per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animal were observed frequently after administration of the test substance and at least once a day for 14 days in order to determine the reversibility of any clinical signs. The appearance or disappearance of any clinical signs was recorded for each animal. Mortality was checked frequently just after administration of the test substance and at least twice a day during the 14-day observation period. The animal were individually weighed just before administration of the test substance and then on days 5, 8 and 15. The body weight gain of the treated animals was compared to a reference curve of the C.I.T. control animals with the same initial weight.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

not applicable

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 500 - < 1 000 mg/kg bw

Based on:

test mat.

Mortality:

There were 0/5, 3/5 and 3/5 deaths in male between 30 minutes and 2 hours after treatment at 500, 1000 and 1500 mg/Kg respectively. In females, there were 1/5, 4/5 and 4/5 mortalities between 1 and 4 hours after treatment at 500, 1000 and 1500 mg/Kg respectively.

Clinical signs:

At 500, 1000 and 1500 mg/Kg, the principal clinical signs of toxicity observed after 15 or 30 minutes were sedation and dyspnea. In a few animals, tonico-clonic convulsions were seen. At 1000 and 1500 mg/Kg, hypersalivation was observed. The behaviour of the surviving animals was normal between 2 to 3 days post-treatment. At 1000 and 1500 mg/Kg, orange coloured uro-genital area and orange coloured sawdust were noted during 48 hours. At 500 mg/Kg, orange colouration of sawdust was observed during 48 hours.

Body weight:

The body weight gain of the surviving animals was similar to historical controls between days 1 and 15.

Gross pathology:

Macroscopic post-mortem examination revealed no abnormalities in the animals (3 animals at 1000 and 1500 mg/Kg and 9 animals at 500 mg/Kg) sacrified at the end of the study. dark red area of the stomach and intestines was observed at necropsy of 1 animal treated at 1000 mg/Kg and at necropsy of 7 animals treated at 1500 mg/Kg. Orange colouration of the tissues and organs was noted in all animals dead during the study.

Any other information on results incl. tables

Clinicalsigns

Dose (mg/Kg)	Time	Animals		Clinical signs
		Males	Females
500	15 min	01 -02 -03 -04 -05	01 -02 -03 -04 -05	None
	30 min	04   01-02-03-05	05   01-02-03-04	Tonico-clonic convulsions Sedation, dyspnea
	1 h	04     01-02-03-05	04-05     01-02-03	Tonico-clonic convulsions, orangecolourationof the extremities Sedation, dyspnea, orangecolourationof the extremities
	2 h	04     01-02-03-05	04-05     01-02-03	Tonico-clonic convulsions, orangecolourationof the extremities Sedation, dyspnea, orangecolourationof the extremities
	4 h	01-02-03-04-05	04 01-02-03-05	Mortality Sedation, piloerection, dyspnea, orangecolourationof the extremities
	D3 to D15	01-02-03-04-05	01-02-03-05	None
1000	15 min	02   01-03-04-05	01-03-04   02-05	Convulsions,dyspnea, hypersalivation Sedation,dyspnea, hypersalivation
	30 min	03 02-05   01-04	01-02-03-04-05	Mortality Tonico-clonic convulsions, dyspnea,hypersalivation Sedation, dyspnea, ataxia,hypersalivation
	1 h	02 01-04-05	01-02-04-05 03	Mortality Tonico-clonic convulsions, dyspnea, orangecolourationof the extremities
	2 h	01 04     05	03	Mortality Tonico-clonic convulsions, dyspnea, orangecolourationof the extremities Sedation, dyspnea, orangecolourationof the extremities
	4 h	04-05	03	Sedation, dyspnea, orangecolourationof the extremities Hypokinesia, orangecolourationof the extremities
	D 2 to D 15	04-05	03	None
1500	15 min	02-03   01-04-05	01-04   02-03-05	Tonico-clonic convulsions,dyspnea, hypersalivation Sedation,dyspnea,tremors, hypersalivation
	30 min	01-02-03-04-05	01-02-03-04-05	Tonico-clonic convulsions, dyspnea,hypersalivation
	1 h	02-03-05 01-04	01-04-05 02-03	Mortality Tonico-clonic convulsions, dyspnea, orangecolourationof the extremities
	2 h	01-04	03 02	Mortality Sedation, dyspnea, orangecolourationof the extremities
	4 h	01-04	02	Sedation, dyspnea, orangecolourationof the extremities
	D 2	01-04	02	Hypokinesia
	D 3 to D 15	01-04	02	None

 

Bodyweightgain (g)

Dose	Volume	Sex		Days
mg/Kg	mL/Kg			1 to 5	5 to 8	8 to 15
500	10	Male	M	52	32	61
			SD	6	5	8
1000	10		M	39	32	58
			SD	14	6	1
1500	10		M	49	30	46
			SD	11	4	15
500	10	Female	M	34	8	31
			SD	4	7	4
1000	10		M	36	7	30
			SD
1500	10		M	30	12	34
			SD

 

M : mean

SD : standard deviation

Animal found dead during the study not mentioned

 

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Death were mainly noted between 1 and 2 hours post-treatment. Mortality rates were 10%, 70% and 70% in rats at 500, 1000 and 1500 mg/Kg respectively. No sex-related difference was noted.
The body weight gain of the surviving animals was not influenced by the treatment.
Macroscopic post-mortem examination revealed no abnormalities in the animals sacrificed at the end of the study. Orange or dark red colouration of the tissues and organs was observed in all animals found dead during the study at all dose levels.
Under these experimental conditions, the LD50 of the test item administered by oral route in rats was higher than 500 mg/Kg and less than 1000 mg/Kg.

Executive summary:

This GLP-compliant study was performed to assess the acute oral toxicity of the test item, according to fixed dose OECD Guideline 401 method (dated 24th February 1987).

The test substance was administered by gavage to 3 groups of 10 fasted Sprague-Dawley rats (5 males and 5 females) at the dose levels of 500, 1000 and 1500 mg/Kg. The test item was administered in suspension  in 1,2 -propanediol, 99% at a volume of 10 mL/Kg. They were observed for mortality, clinical signs and bodyweight for 14 days observation period and were subjected to necropsy at day 15 for macroscopic examination of the main organs.

Death were mainly noted between 1 and 2 hours post-treatment. Mortality rates were 10%, 70% and 70% in rats at 500, 1000 and 1500 mg/Kg respectively. No sex-related difference was noted.

The body weight gain of the surviving animals was not influenced by the treatment.

Macroscopic post-mortem examination revealed no abnormalities in the animals sacrificed at the end of the study. Orange or dark red colouration of the tissues and organs was observed in all animals found dead during the study at all dose levels.

Under these experimental conditions, the LD50 of the test item administered by oral route in rats was higher than 500 mg/Kg and less than 1000 mg/Kg.

Thus, according to GHS regulation, the test substance is classified as hazardous category 4 according to GHS regulation.