Benzoic acid, 2-[[(2'-cyano[1,1'-biphenyl]-4-yl)methyl]amino]-3-nitro-, methyl ester

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Step 1: Date of start of the experimental period: 25 May 2004 - Step 2: Date of end of the experimental period: 4 Jun. 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

methyl 2-[[4-(2-cyanophenyl)phenyl]methylamino]-3-nitrobenzoate

EC Number:

604-135-1

Cas Number:

139481-28-0

Molecular formula:

C22H17N3O4

IUPAC Name:

methyl 2-[[4-(2-cyanophenyl)phenyl]methylamino]-3-nitrobenzoate

Test material form:

solid: particulate/powder

Details on test material:

batch No. February 2004

Specific details on test material used for the study:

One sample of CAN 3, batch No. February 2004, was used. CAN 3 was stored at ambient temperature and protected from light in accordance with the Sponsor' s instructions.
Appearance: Yellow powder.

Test animals

Species:

rabbit

Strain:

other: New Zealand Albino

Details on test animals or test system and environmental conditions:

Choice of species:
The rabbit was chosen because of its acceptance as a predictor of irritant/corrosive effects of chemicals in man and the recognition by regulatory authorities that this species is suitable for skin irritation studies.

Sex:
Female

Origin:
Animals came from the CEGAV specialised breeding establishment (Les Hautes Noes, Saint Mars d'Egrenne, 61350 Passais la Conception, France).

Date of delivery:
19 May 2004.

Identification:
Animals were individually identified by an ear clip.

Body weight:
Generally between 2.5 kg and 2.8 kg at the start of the experiment.

Acclimatisation:
For at least 5 days before the treatment, in the area where the experiment took place.

Housing:
Animals were housed individually in cages of standard size. Excreta were removed by unrolling plasticised brown paper, previously placed under cages. These cages were placed in an airconditioned (17- 21 °C) animal house kept at between 45% and 65% relative humidity in which non-recycled filtered air was changed approximately ten times per hour. The artificial day/night cycle was 12 hours light and 12 hours darkness with light on at 7.30 a.m.

Feeding:
SDS/DIETEX STANRAB (P) SQC feed was distributed daily at fixed times. The criteria for acceptable levels of contaminants in the feed supply was within the limits of the analytical specifications established by the diet manufacturer. The certificate of analysis concerning this feed product is included in the study report

Drinking water:
Drinking water was available ad libitum in polycarbonate feeder bottles with a stainless steel nipple. A specimen of water is obtained every 6 months and sent to the Laboratoire Départemental d'Analyse du Cher- 216 rue Louis Mallet, 18014 Bourges Cedex, France- for analysis.
The criteria for acceptab1e levels of contaminants in the water supply was within the limits of the
analytical specifications.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

yes

Amount / concentration applied:

As required by the guideline, 0.5 g of CAN 3 was applied. The test substance was placed on a ganze square of approximately 6 cm2 . Bach application was
sufficiently moistened with 0.5 mL of sterile water in order to allow good contact between the test substance and the skin. The side of the ganze square covered with the test substance was applied to the skin. This ganze square was protected by a pad consisting of a semi-occlusive micro-porous and non-allergenic dressing and held in piace with a supple and aerated fastening system consisting of elastic, adhesive, hypoallergenic tape. The animals were fitted with this pad for the duration of the exposure period.

Duration of treatment / exposure:

CAN 3 was applied once only, at a duration of exposure of 4 hours.

Observation period:

One hour, 24 hours, 48 hours and 72 hours approximately after remaval of the dressing, any skin lesions, which may bave developed on the right fiank of each animai, were evaluated.

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: the test substance was placed on a gauze square of approximately 6 cm2.
- % coverage:
- Type of wrap if used: The side of the gauze square covered with the test substance was applied to the skin. This gauze square was protected by a pad consisting of a semi-occlusive micro-porous and non-allergenic dressing and held in place with a supple and aerated fastening system consisting of elastic, adhesive, hypoallergenic tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period, the dressing was removed and residual test substance removed, using sterile water. Any damage to the epidermis was avoided.
- Time after start of exposure: CAN 3 was applied once only, at a duration of exposure of 4 hours.

OBSERVATION TIME POINTS
One hour, 24 hours, 48 hours and 72 hours approximately after remaval of the dressing, any skin lesions, which may have developed on the right flank of each animal, were evaluated.

SCORING SYSTEM:
- Method of calculation: Lesions were graded using the following scoring system in accordance with OECD Guideline No. 404.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 5

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 6

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Any other information on results incl. tables

The application of the test substance induced yellow coloration of the application site. In order to allow satisfactory conditions for grading, the application site was rinsed using a waterlogged compress at the time of removal of the dressing. The absence of local reactions to the rinsing procedure with sterile water was confirmed simultaneously: no oedema and no erythema was noted at the issue of this procedure. Any cutaneous lesion was evaluated approximately one hour, 24, 48 and 72 hours after removal of the dressing. Mean indices were calculated from results obtained from each rabbit at times 24, 48 and 72 hours.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions adopted, CAN 3 (batch February 2004) was found to be non irritant for the skin of the Rabbit.

Executive summary:

Irritant property and/or degree of corrosion of CAN 3 (batch February 2004) were evaluated in the Rabbit following a single semi-occluded application to intact skin, in compliance with requirements of OECD Guideline No, 404 (April 24, 2002) and Method B4 of Council Directive No, 67/548/EEC and subsequent amendments. Three animals were used for the study.

0.5 g of CAN 3 was applied on the skin. Adjacent surfaces of untreated skin served as a control for the trial. The test substance was moistened with 0.5 mL of sterile water in order to allow good contact between the test substance and the skin. Semi-occlusive dressings held the test substance in place for 4 hours on the skin of the animals. The application of the test substance induced yellow coloration of the application site. In order to allow satisfactory conditions for grading, the application site was rinsed using a waterlogged compress at the time of removal of the dressing. The absence of local reactions to the rinsing procedure with sterile water was confirmed simultaneously: no oedema and no erythema was noted at the issue of this procedure. Any cutaneous lesion was evaluated approximately one hour, 24, 48 and 72 hours after removal of the dressing. Mean indices were calculated from results obtained from each rabbit at times 24, 48 and 72 hours.