Tetrasodium 4,4'-bis[[4-morpholino-6-(p-sulphonatoanilino)-1,3,5-triazin-2-yl]amino]stilbene-2,2'-disulphonate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 April 2008 to 2 May 2008.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted Apri124, 2002

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Tetrasodium 4,4'-bis[[4-morpholino-6-(p-sulphonatoanilino)-1,3,5-triazin-2-yl]amino]stilbene-2,2'-disulphonate

EC Number:

249-323-0

EC Name:

Tetrasodium 4,4'-bis[[4-morpholino-6-(p-sulphonatoanilino)-1,3,5-triazin-2-yl]amino]stilbene-2,2'-disulphonate

Cas Number:

28950-61-0

Molecular formula:

C40H40-xN12NaxO14S4

IUPAC Name:

tetrasodium 2,2'-ethene-1,2-diylbis[5-({4-morpholin-4-yl-6-[(4-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)benzenesulfonate]

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: 0582900
- Expiration date of the lot/batch: 25-FEB-2013

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature (range of 20 ± 5 °C), light protected.
- Stability under storage conditions: Stable under storage conditions.

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Netherlands BV Kreuzelweg 53 NL-596l NM Horst / The Netherlands Postbus 6174 NL-5960 AD Horst / The Netherlands
- Age at study initiation: 15 weeks (male) and 14 weeks (females).
- Allocation: Male No. 61, Female Nos. 62, 63.
- Diet: pelleted standard Provimi Kliba 3418 rabbit maintenance (batch no. 03/08) provided by Provimi Kliba AG, CH-4303 Kaiseraugst ad libitum
- Water: community tap water from Füllinsdorf ad libitum
- Acclimation period: under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature: continuously monitored environment with ranges for room temperature 17-23 °C.
- Humidity: between 30-70 % (values above 70 % during cleaning process possible).
- Air changes: air-conditioned with 10-15 air changes per hour.
- Photoperiod: automatically controlled light cycle of 12 hours light and 12 hours dark.
- Other: music during the daytime light period.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g (per animal) of test item was weighed as delivered by the Sponsor and then moistened with approximately 0.5 ml of purified water before application.
- Concentration: a formulation of a 1 % (w/w) solution in purified water was prepared. The pH was found to be 7-8.

Duration of treatment / exposure:

4 hours

Observation period:

The skin reaction was assessed at approximately 1, 24, 48 and 72 hours, after exposure (removal of the dressing, gauze patch and test item).

Number of animals:

Three (one male and two females)

Details on study design:

TEST SITE
- Area of exposure: four days before treatment, the left flank was clipped with an electric clipper, exposing an area of approximately 100 cm2 (10 cm x 10 cm). The skin of the animals was examined one day before treatment, and regrown fur of all animals was clipped again. Animals with overt signs of skin injury or marked irritation which may have interfered with the interpretation of the results were not used in the test.
- Application: on the day of treatment, 0.5 g of test item was placed on a surgical gauze patch (ca. 2.5 cm x 2.5 cm). This gauze patch was applied to the intact skin of the clipped area. The patch was covered with a semi-occlusive dressing.
- Type of wrap if used: the dressing was wrapped around the abdomen and anchored with tape.

REMOVAL OF TEST SUBSTANCE
- Washing: the dressing was removed after 4 hours and the skin was flushed with lukewarm tap water to clean the application site.
Necropsy
No necropsy was performed on the animals sacrificed at termination of observation.
All rabbits were sacrificed by an intravenous injection of Pentobarbitone into the ear vein at a dose of at least 1 ml/kg body weight and discarded.

SCORING SYSTEM
The skin reaction was assessed according to the numerical scoring system listed in the Commission Directive 2004173/EC, April 29, 2004.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Results 2

Irritation parameter:

edema score

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0 - 2.3

Max. score:

4

Irritant / corrosive response data:

A slight to well-defined erythema was observed in all animals 1-hour after test item exposure and persisted as slight up to the 24-hour reading. No abnormal findings were observed on the treated skin of any animal 48 hours after treatment.
Coloration: no staining produced by the test item of the treated skin was observed.
Corrosion: neither alterations of the treated skin were observed nor were corrosive effects evident on the skin.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
The body weights of all rabbits were considered to be within the normal range of variability.

Any other information on results incl. tables

Individual animal skin irritation scores

Animal N.	Sex	Parameter	1 h	24 h	48 h	72 h	Mean*
61	M	Erythema	1	1	0	0	0.33
		Oedema	0	0	0	0	0.00
62	F	Erythema	2	1	0	0	0.33
		Oedema	0	0	0	0	0.00
63	F	Erythema	2	1	0	0	0.33
		Oedema	0	0	0	0	0.00

* Time point: 24, 48 and 72 hrs after patch removal.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 66042/A is not considered as irritating to rabbit skin.

Executive summary:

The primary skin irritation potential of test item was investigated according to OECD test guideline no. 404. The test item was applied by topical semi-occlusive application of 0.5 g to the intact left flank of each of three young adult New Zealand White rabbits. The duration of treatment was four hours. The scoring of skin reactions was performed 1,24, 48 and 72 hours after removal of the dressing. The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately.

The mean erythema/eschar score of the three animals was 0.33 for all animals and the mean oedema score was 0.00 for all animals. The application of test item to the skin resulted in very slight to well-defined erythema. This effect was reversible and was no longer evident 48 hours after treatment. The test item caused no staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no clinical signs were observed. Based on the study results, FAT 66042/A is considered as not irritating to rabbit skin.