3,10-diamino-6,13-dichloro-2-((6-(((4-(1,1-dimethylethyl)phenyl)sulfonyl)amino)-2-naphthalenyl)sulfonyl)-4,11-triphenodioxazinedisulfonic acid, lithium potassium sodium salt

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October 14, 2001 - December 04, 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

July 31, 1992.

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted July 17, 1992.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

440-770-9

EC Name:

-

Cas Number:

371921-63-0

Molecular formula:

C38 H29 Cl2 N5 O12 S4 .x K .x Li .x Na

IUPAC Name:

3,10-diamino-2-{[6-(4-tert-butylbenzenesulfonamido)naphthalen-2-yl]sulfonyl}-6,13-dichloro-5,12-dioxa-7,14-diazapentacene-4,11-disulfonic acid lithium hydride potassium hydride sodium hydride

Test material form:

solid

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage Scientifique des Dombes F-01400 Chatillon sur Chalaronne I France
- Age at study initiation: 16-17 weeks (male #31); 11-12 weeks (female #32); 18-19 weeks ( female #33)
- Weight at study initiation: 2979g (male #31); 2105g (female #32); 3339g ( female #33)
- Housing: Individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with wood (RCC Ltd, Fullinsdorf) and haysticks for gnawing.
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 91 /01) provided by Provimi Kliba AG, CH-4303 Kaiseraugst. Results of analysis are archived at RCC Ltd, ltingen.
Haysticks (QS no. 24/01) provided by Eberle Nafag AG, CH-9200 Gossau.
- Water (e.g. ad libitum): Community tap water from Fullinsdorf, ad libitum. Results of
bacteriological, chemical and contaminant analyses are archived at RCC Ltd, ltingen.
- Acclimation period: 5 days (October 2001, 16 to 21). Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
Amount(s) applied (volume or weight with unit): 0.5g test item/animal

VEHICLE
The test item was moistened with approximately 0.1 ml purified water before application.

Duration of treatment / exposure:

4 hours

Observation period:

14 days

Number of animals:

1 male & 2 females

Details on study design:

TEST SITE
- Area of exposure: 4 cm x 4 cm
- % coverage: 16% (are of patch on which th e test item is applied / area of left flank that is clipped)
- Type of wrap if used: Test item was placed on a surgical gauze patch (ca. 4 cm x 4 cm). This gauze patch was applied to the intact skin of the clipped area. The patch was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site so that any reactions (erythema) were clearly visible at that time.
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
(indicate if minutes, hours or days): h, 24h, 48h, 72h, 7d, 10d, 14d

SCORING SYSTEM:
- Method of calculation: Grading of Skin Reactions (ERYTHEMA AND ESCHAR FORMATION & OEDEMA FORMATION) from EEC COMMISSION DIRECTIVE 92/69/EEC, JULY 31, 1992. Note that the same grading is used in OECD guideline 404 (adopted: 28 July 2015)

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal: #31, #32, #33

Time point:

other: 1h, 24h, 48h, 72h, 7d, 10d, 14d

Score:

0

Max. score:

0

Reversibility:

other: not relevant in absence of effects

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

edema score

Basis:

animal: #31, #32, #33

Time point:

other: 1h, 24h, 48h, 72h, 7d, 10d, 14d

Score:

0

Max. score:

0

Reversibility:

other: not relevant in absence of effects

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No abnormal findings were noted for erythema, oedema and flaking for eachanimal at each time point.

Other effects:

- Other adverse local effects: light blue staining produced by the test item is observed for each animal at each time point.

Any other information on results incl. tables

VIABILITY/MORTALITY/CLINICAL SIGNS

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.

IRRITATION

The test item did not elicit any skin reactions at the application site of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0.

COLORATION

A light blue coloured staining was observed at the test site of all animals throughout the entire 14 day observation period.

CORROSION

No irreversible alterations of the treated skin were observed nor were corrosive effects evident on the skin.

BODY WEIGHTS

The body weights of all rabbits were considered to be within the normal range of variability.

Animal#	Sex	First Day of Acclimatization	Day of Treatment	Last Day of Observation
31	Male	2979g	3057g	3317g
32	female	2105g	2307g	2690g
33	female	3339g	3443g	3675g

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on criteria defined in Regulation (EC) No 1272/2008 with the exception of the blue staining which persisted in all animals up to termination of the test, BLUE GS 5664.80 is considered to be "not irritating" to rabbit skin and is not classified according to CLP criteria.

Executive summary:

The primary skin irritation potential of BLUE GS 5664.80 was investigated by topical semiocclusive 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, as well as 7, 10 and 14 days after removal of the dressing.

The test item did not elicit any skin reactions at the application site of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0.

A light blue coloured staining was however observed at the test site of all animals throughout the entire 14 day observation period. No corrosive effects were noted on the treated skin of any animal at any measuring interval.

Based upon the referred classification criteria (EEC Commission Directive 93/21/EEC of April 27, 1993 and Regulation (EC) No 1272/2008), with the exception of the blue staining which persisted in all animals up to termination of the test, BLUE GS 5664.80 is considered to be "not irritating" to rabbit skin and is not classified according to CLP criteria.