4-amino-3-nitrophenol

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 1992-03-05 to 1992-05-25

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

The test item is 6% diluted substance tested in vehicle instead of smallest amount of water (or, where necessary, of another suitable vehicle) in according to 404 OECD (adopted: 28 July 2015), no analytical certificate of the test substance available.

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 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

12 May 1981

Deviations:

yes

Remarks:

yes The test substance is 6% diluted in vehicle. However, according to 404 OECD protocols (28 July 2015), a solid test substance has to be used with the smallest amount of water (or, where necessary, another suitable vehicle).

GLP compliance:

yes

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: flakes

Remarks:

bown red flakes

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: not specified
- Purity test date: not specified
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature
- Stability under test conditions: no data
- Solubility and stability of the test substance in the solvent/vehicle: no data
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: no data
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test substance was ground down to a fine dust and then suspended at a concentration of 6% in the 1,2-propanediol
- Final dilution of a dissolved solid, stock liquid or gel: 6% (w/w)

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage Cunicole du Val de Selle, France
- Age at study initiation: not specified
- Weight at study initiation: 2.4±0.05kg
- Housing: Individually in polystyrene cages (35 x 55 x 32 cm or 48.2 x 58 x 36.5 cm)
- Diet (e.g. ad libitum): Certified pellet diet "Rabbits sustenance ref. 112C"(U.A.R., 91360 Villemoisson-sur-Orge, France). An analytical certificate of the quality of the food and the major food contaminants (pesticides, heavy metals, mycotoxins, etc.) was made available by the supplier and given for each batch.
- Water (e.g. ad libitum): Free access to tap water filtered by a 0.22 micron filtered membrane (Société Millipore, 78140 Velizy, France) and contained in water bottles. Bacteriological and chemical analyses of the water and detection of the major contaminants (pesticides, heavy metals and nitrosamines) were made periodically (laboratoire Municipal et Regional de Rouen, 76000 Rouen, France- Centre de nutrition humaine , 54000 Nancy France - Laboratoire Departemental d'analyses, 27000 Evreux, France)
- Acclimation period: at least 5 days during which they were observed daily

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18±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: 21 March 1992 To: 29 March 1992

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

other: 1,2-propanediol

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 ml
- Concentration (if solution): 6%
VEHICLE
- Amount(s) applied (volume or weight with unit): 0.47 ml
- Concentration (if solution): 94%
- Lot/batch no. (if required): 1256840
- Purity:99%

Duration of treatment / exposure:

4 Hours

Observation period:

One hour, 24h, 48h, 72h

Number of animals:

3 males

Details on study design:

TEST SITE
- Preparation : The day before treatment, the flancks of each animal were partially clipped with an electric clipper. Only animals showing no obvious signs of irritation were used for the study.
- Application : A single dose of 0.5 ml of a solution containing the test substance at a concentration of 6% in 1.2-propanediol was applied to a 6cm² dry hydrophilic gauze patch (Semes France, France) and this was then applied to the right flank of each animal.
- Area of exposure: 6 cm²
- Type of wrap if used: adhesive hypoallergic aerated semi-occlusive dressing (Laboratoires de Pansements et d'Hygiène, France)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the dressing were removed and residual test substance was removed with a gauze patch saturated with water for injections batch n° 1343 (Biosédra, France).
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
1, 24, 48, 72 hours

SCORING SYSTEM:
- Method of calculation:
Oedema Formation
No oedema ..........................................................................................................................0
Very slight oedema (barely perceptible)..............................................................................1
Slight oedema (edges of area well defined by definite raising) ..........................................2
Moderate oedema (raised approximately 1millimetre) ..........................................................3
Severe oedema (raised more than 1 millimetre and extended beyond area of exposure) ..4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

edema score

Basis:

mean

Remarks:

Mean calculated score for each site

Time point:

other: 1/24/48/72H

Score:

0

Max. score:

4

Results 2

Irritation parameter:

erythema score

Basis:

mean

Remarks:

Mean calculated score for each site

Time point:

other: 1/24/48/72H

Max. score:

4

Remarks on result:

not determinable

Remarks:

Possible erythema was obscured by colouration caused by the residual test substance.

Irritant / corrosive response data:

Macroscopic examination: no edema was noted. Possible erythema was obscured by colouration caused by the residual test substance.
Microscopic examination: slight dermal inflammatory cell aggregation was noted in 1 animal. This type of lesion can be found spontaneously in the laboratory rabbit and is considered to be of no toxicological importance.

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

Under the conditions of this study, the test item at 6% in 1.2-propanediol caused no edema to rabbit skin and no lesions related to the application of the test item were noted at the microscopic examination. The study cannot be used for classification. Indeed, the test dose is 6% in diluted vehicle instead of pure or with the smallest amount of water (or,where necessary, of another suitable vehicle) according to REACh and CLP regulation.

Executive summary:

This GLP-compliant study was performed to assess the potential of the test material to induce skin

irritation in vivo, according to OECD Guideline 404 (dated May 12, 1981). 0.5 ml of 6% test substance diluted in 1.2 propanediol were applied under semi-occlusive dressing during 4 hours, on a shaved skin area (right flank) of 3 male rabbits. The left flank served as a control. Skin reactions were recorded 1 hour and then 24, 48 and 72 hours after the removal of the dressing. Results: Macroscopic examination: no edema was noted. Possible erythema was obscured by colouration caused by the residual test substance. Microscopic examination: slight dermal inflammatory cell aggregation was noted in 1 animal. This type of lesion can be found spontaneously in the laboratory rabbit and is considered to be of no toxicological importance.

The study cannot be used for classification. Indeed, the test dose is 6% in diluted vehicle instead of pure or with the smallest amount of water (or, where necessary, of another suitable vehicle) according to REACh and CLP regulation.