Trisodium bis[(3'-nitro-5'-sulfonato-(6-amino-2-[4-(2-hydroxy-1-naphtylazo)phenylsulfonylamino]pyrimidin-5-azo)benzene-2',4-diolato)]chromate(III)

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May 19, 1994 - May 26, 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1992

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

1992

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium bis[(3'-nitro-5'-sulfonato-(6-amino-2-[4-(2-hydroxy-1-naphtylazo)phenylsulfonylamino]pyrimidin-5-azo)benzene-2',4-diolato)]chromate(III)

EC Number:

418-220-4

EC Name:

Trisodium bis[(3'-nitro-5'-sulfonato-(6-amino-2-[4-(2-hydroxy-1-naphtylazo)phenylsulfonylamino]pyrimidin-5-azo)benzene-2',4-diolato)]chromate(III)

Molecular formula:

C52H32CrN18Na3O20S4

IUPAC Name:

chromium(3+) trisodium bis(6-amino-2-{4-[2-(2-hydroxynaphthalen-1-yl)diazen-1-yl]benzenesulfonamido}-5-[2-(3-nitro-2-oxido-5-sulfonatophenyl)diazen-1-yl]pyrimidin-4-olate)

Test material form:

solid: particulate/powder

Test animals

Species:

rabbit

Strain:

other: New Zealand

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Animals: rabbit, Chbb: NZW (SPF)
- Source: Dr. Karl Thomae GmbH Postfach 1755 D-88397 Biberach an der Riss.
- Age at start of treatment: males at 13 weeks, females at 12 weeks.
- Body weight at start of acclimatization: male at 2.6 kg, females at 2.1 – 2.5 kg.
- Body weight at start of treatment: male at 2.7 kg, females at 2.2 – 2.7 kg.
- Identification: by unique cage number and corresponding tags.
- Acclimatization: four days under test conditions after health examination. Only animals without any visual signs of illness were used for the study.
- Accomodation: individually in stainless steel cages with an automatic cleaning system equipped with feed hoppers, drinking water bowls and wood for gnawing.
- Diet: pelleted standard Kliba 341, Batch 83/94 rabbit maintenance diet ("Kllba", Klingentalmueble AG, CH-4303 Kaiseraugst), ad libitum.
- Water: community tap water from Itingen, ad libitum. A bacteriological, chimical and contaminant analyses were performed.

ENVIRONMENTAL CONDITIONS
- Temperature: 20 ± 3 °C
- Humidity: 40-70 %
- Air changes: 10-15 ACH
- Photoperiod: 12 hours artificial fluorescent light (approx 100 Lux) / 12 hours dark, misic during light period.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Remarks:

bi-distilled

Controls:

yes, concurrent no treatment

Amount / concentration applied:

- Amount applied: 0.5 g/animal; test article applied moistoned.
- Application area: 6 cm2

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

1 male and 2 females

Details on study design:

TREATMENT
The skin of the animals was examined once, 1 day prior to test article administration. Only those animals with no signs of skin injury or irritation were used in the test.

TEST SITE
- Area of exposure: approximately 24 hours before treatment, the dorsal fur was clipped w1th an electric clipper, exposing an area of approximately 100 square centimetres (10 cm X 10 cm).
- Type of wrap if used: area exposed was covered with a 3.0 cm x 3.0 cm patch of surgical gauze. The gauze was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with an elastic bandage. The test article was applied in such a position as to preclude oral ingestion after removal of the dressing.

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

SCORING SYSTEM
The skin reaction was assessed at 1, 24, 48 and 72 hours after the removal of the dressing, gauze patch and test article.
The possible corrosive properties of the test article and the staining of the treated skin area related to the test article were described and recorded.
The skin reaction was assessed according to the following numericaí scoring system.

Erythema and eschar formation
No erythema 0
Very sllght erythema (barely perceptible) 1
Well defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) to sllght eschar formation (injuries in depth) 4
Maximum possible score 4

Edema formation
No edema 0
Very slight edema (barely perceptible) 1
Sllght edema (edges of área well defined by definite raising) 2
Moderate edema (raised approximately 1 mm) 3
Severe edema (raised more than 1 mm and extending beyond area of exposure) 4
Maximum possible score 4

Maximum cumulative score: 8

OTHER OBSERVATIONS
Viability, mortality and clinical signs: daily during observation period.
Body weights: at the start of acclimatization, on day 1 of test (application day) and at termination of observation.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Reversibility:

fully reversible

Results 2

Irritation parameter:

edema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Reversibility:

fully reversible

Irritant / corrosive response data:

Irritation
Test item showed a primary irritation score of 0.89 (max. 8.0) when applied to healthy intact rabbit skin.
Local signs (mean values from 24 to 72 hours) consisted of grade 0.44 erythema and grade 0.44 edema.

After 1 hour: skin red coloured by test article
After 24 hours: skin light-red coloured by test article; well-defined erythema and slight edema (animal ID 8 and 9, females)
After 48 hours: skin light-red coloured by test article; very slight erythema and very slight edema (animal ID 8, female)
After 72 hours: skin light-red coloured by test article; no other abnormal changes visible

Coloration
In the area of application red staining of the treated skin by pigment or colouring of the test article was observed.

Corrosion
No destructions or irreversible alterations of the treated skin were observed.
No corrosive effect was evident on the skin.

Other effects:

Clinical signs and mortality
No systemic symptoms were observed in the animals during the test and observation period, and no mortality occurred.

Body weights
The body weight gain of all rabbits was within the normal range of variability.

Any other information on results incl. tables

Individual mean score at 24, 48 and 72 hours

Reaction	Time	Animal/sex
		7 M	8 F	9 F
Erythema	1 hr	0	0	0
	24 hrs	0	2	1
	48 hrs	0	1	0
	72 hrs	0	0	0
Mean 24, 48 and 72 hrs		0	1	0.33
Edema	1 hr	0	0	0
	24 hrs	0	2	1
	48 hrs	0	1	0
	72 hrs	0	0	0
Mean 24, 48 and 72 hrs		0	1	0.33

Skin irritation score

Time period	Animal ID	Sex	Erythema	Edema	Comulative score	Mean cumulative score
1 h	7	M	0	0	0	0.00
	8	F	0	0	0
	9	F	0	0	0
24 h	7	M	0	0	0	2.00
	8	F	2	2	4
	9	F	1	1	2
48 h	7	M	0	0	0	0.67
	8	F	1	1	2
	9	F	0	0	0
72 h	7	M	0	0	0	0.00
	8	F	0	0	0
	9	F	0	0	0

Applicant's summary and conclusion

Interpretation of results:

other: not classified according to the CLP Regulation (EC 1272/2008)

Conclusions:

The substance was found to be non-irritant to the skin of rabbits under the test conditions.

Executive summary:

The skin irritation potential of the test material was investigated by topical application of 0.5 g onto 6 cm² intact dorsal skin of each of three young adult New Zealand rabbits. The scoring of skin reactions was performed 1, 24, 48 and 72 hours after removal of the dressing. The scores of each animal at the following reading times (24, 48, 72 hours) was used in calculating the respective mean values for each type of lesion.

The mean values 24/48/72 h for both erythema and oedema were 0, 1 and 0.33 in animals #1, #2 and #3 respectively. All the effects observed were reversible within the 72 h-study period.