1,7-Naphthalenedisulfonic acid, 4-amino-3-[(1E)-2-(2,4-disulfophenyl)diazenyl]-5-hydroxy-6-[(1E)-2-(4-nitro-2-sulfophenyl)diazenyl]-, sodium salt (1:5)

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13 July 2015 - 16 Aug 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed at a GLP laboratory in accordance with prescribed OECD, EU, US and Japanese test guidelines.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

Deviations from the maximum level of daily mean relative humidity occurred. Laboratory historical data do not indicate an effect of the deviations. The study integrity was not adversely affected by the deviation.

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Deviations from the maximum level of daily mean relative humidity occurred. Laboratory historical data do not indicate an effect of the deviations. The study integrity was not adversely affected by the deviation.

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

yes

Remarks:

Deviations from the maximum level of daily mean relative humidity occurred. Laboratory historical data do not indicate an effect of the deviations. The study integrity was not adversely affected by the deviation.

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: JMAFF Guidelines (2000), including the most recent revisions.

Deviations:

yes

Remarks:

Deviations from the maximum level of daily mean relative humidity occurred. Laboratory historical data do not indicate an effect of the deviations. The study integrity was not adversely affected by the deviation.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

pentasodium 4-amino-3-[(1E)-2-(2,4-disulfonatophenyl)diazen-1-yl]-5-hydroxy-6-[(1E)-2-(4-nitro-2-sulfonatophenyl)diazen-1-yl]naphthalene-1,7-disulfonate

EC Number:

811-858-8

Cas Number:

2149571-68-4

Molecular formula:

C22 H11 N6 O18 S5 .5Na

IUPAC Name:

pentasodium 4-amino-3-[(1E)-2-(2,4-disulfonatophenyl)diazen-1-yl]-5-hydroxy-6-[(1E)-2-(4-nitro-2-sulfonatophenyl)diazen-1-yl]naphthalene-1,7-disulfonate

Constituent 2

Reference substance name:

pentasodium 4-amino-3-[(2,4- disulfonatophenyl)diazenyl]-5-hydroxy-6-[(4-nitro-2- sulfonatophenyl)diazenyl]naphthalene-1,7-disulfonate

IUPAC Name:

pentasodium 4-amino-3-[(2,4- disulfonatophenyl)diazenyl]-5-hydroxy-6-[(4-nitro-2- sulfonatophenyl)diazenyl]naphthalene-1,7-disulfonate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): K1600 black dye
- Physical state: Powder, black

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River France, L’Arbresle Cedex, France
- Age at study initiation: Between 12 and 24 weeks old
- Weight at study initiation: At least 1.5 kg.
- Housing: Individually housed in labeled cages with perforated floors (Ebeco, Germany, dimensions 67 x 62 x 55 cm) and shelters (Ebeco, Germany, dimensions 40 x 32 x 23 cm).
- Diet (e.g. ad libitum): Pelleted diet for rabbits (Global Diet 2030 from Harlan Teklad®, Mucedola, Milanese, Italy) approximately 100 grams per day.
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C,
- Humidity (%): a relative humidity of 40 to 70%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Remarks:

20-23%

Controls:

other: Adjacent areas of the untreated skin of each animal served as controls.

Amount / concentration applied:

0.5g
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5g
- Concentration (if solution): Test substance was moistened with 0.4 mL of water.

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.4mL used to moisten the test substance.

Duration of treatment / exposure:

Four hours after application.

Observation period:

72 hours.

Number of animals:

3 males

Details on study design:

TEST SITE
- Area of exposure: Dorsal area, 150 square centimeters (10 x 15 cm)
- Type of wrap if used: metalline patch

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The skin cleaned of residual test substance using tap water and watery ethanol (50% v/v).
- Time after start of exposure: 4 hours

SCORING SYSTEM:
The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

ERYTHEMA AND ESCHAR FORMATION:
No erythema: 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beef redness)* 4
* Where signs of necrosis or corrosion (injuries in depth) prevent erythema scoring, the maximum grade for erythema (= 4) is given.


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 1 millimeter) 3
Severe oedema (raised more than 1 millimeter and extending beyond the area of exposure) 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Remarks:

Animal 937

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Max. score:

0

Reversibility:

no data

Results 2

Irritation parameter:

erythema score

Remarks:

Animal 938

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Max. score:

0

Reversibility:

no data

Results 3

Irritation parameter:

erythema score

Remarks:

Animal 939

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Max. score:

0

Reversibility:

no data

Results 4

Irritation parameter:

edema score

Remarks:

Animal 937

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Max. score:

0

Reversibility:

no data

Results 5

Irritation parameter:

edema score

Remarks:

Animal 938

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Max. score:

0

Reversibility:

no data

Results 6

Irritation parameter:

edema score

Remarks:

Animal 939

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Max. score:

0

Reversibility:

no data

Irritant / corrosive response data:

No skin irritation or corrosion was observed duirn the study.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The results were evaluated according to Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 on classification, labelling and packaging of substances and mixtures (including all amendments). No skin irritation was caused by 4 hours exposure to K1600 black dye.

Executive summary:

Study was performed at a GLP laboratory in accordance with prescribed OECD, EU, US and Japanese test guidelines. A minor deviation from the daily mean relative humidity occurred however this deviation was determined not to affect the validity of the study.

Three rabbits were exposed to 0.5 grams of K1600 black dye, moistened with water by application onto clipped skin for 4 hours using a semi-occlusive dressing. Skin reactions were assessed 1, 24, 48 and 72 hours after exposure.

The results were evaluated according to Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 on classification, labelling and packaging of substances and mixtures (including all amendments). No skin irritation was caused by 4 hours exposure to K1600 black dye.