3,3'-[ethylenebis[(3-sulpho-p-phenylene)azo]]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonic acid] hexasodium salt AND 3,3'-[ethylenebis[(3-sulpho-p-phenylene)azo]]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonic] acid, compound with 2,2',2''-nitrilotriethanol (1:6)

Endpoint summary

Description of key information

Three rabbits were dermally treated with the substance during 4 hours. Slight irritation of the skin was seen during the 72 hour observation period. The effects were fully reversible at 72 hours. The substance is not irritating to the skin (Safepharm 2005).

Three rabbits received 96 mg of the substance in the eye. Very slight irritation was seen 1 hour after installation. At the other observations (until 72 hours after instillation) no signs of irritation were found. The substance is not irritating to the eyes (Safepharm 2005).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 July 2005 to 24 July 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study according to the guidelines under GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: David Percival Ltd, Moston, Sandbach, Cheshire UK
- Age at study initiation: 12-20 weeks
- Weight at study initiation: 2.0 to 3.5 kg
- Housing: individually
- Diet: Certified Rabbit Diet (Code 5322) from BCM IPS Ltd, London UK ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

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

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Remarks:

moistened with 0.5 mL of distilled water

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g

Duration of treatment / exposure:

4 hours

Observation period:

72 hours (observations at 1, 24, 48 and 72 hours after removal of the dressing)

Number of animals:

3 males

Details on study design:

TEST SITE
- Area of exposure: 2.5x 2.5 cm2
- Type of wrap if used: cotton gauze patch

REMOVAL OF TEST SUBSTANCE
- Washing (if done): swab with cotton wool soaked in distilled water
- Time after start of exposure: 4 hours

SCORING SYSTEM: Draize

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24-72 h

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24-72 h

Score:

0.44

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Other effects:

Blue staining of the application site at all timepoints

Animal	1		2		3
Time	erythema	oedema	erythema	oedema	erythema	oedema
1	0	0	0	0	0	0
24	1	1	1	1	1	1
48	1	1	1	0	1	0
72	0	0	0	0	0	0

Interpretation of results:

GHS criteria not met

Conclusions:

The substance is not irritating to the skin

Executive summary:

Three rabbits were dermally treated with the substance during 4 hours. Slight irritation of the skin was seen during the 72 hour observation period. The effects were fully reversible at 72 hours. The substance is not irritating to the skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 July 2005 to 04 August 2005

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: David Percival Ltd, Moston, Sandbach, Cheshire UK
- Age at study initiation: 12-20 weeks
- Weight at study initiation: 2.0 to 3.5 kg
- Housing: individually
- Diet: Certified Rabbit Diet (Code 5322) from BCM IPS Ltd, London UK ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

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

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml (96 mg)

Duration of treatment / exposure:

single instillation (eyelids were hold for 1 second)

Observation period (in vivo):

72 hours (observations at 1, 24, 48 and 72 hours after instillation)

Number of animals or in vitro replicates:

3 males

Details on study design:

First one animal was treated. After consideration of the ocular responses of this animal 2 additional animals were treated

REMOVAL OF TEST SUBSTANCE
- Washing (if done): none

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: ophthalmoscope

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24-72 h

Score:

0

Max. score:

0

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24-72 h

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 24-72 h

Score:

0

Max. score:

1

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24-72 h

Score:

0

Max. score:

0

Other effects:

Black staining of the fur arround the treated eye was observed at all timepoints

Animal no	1				2				3
Time	1	24	48	72	1	24	48	72	1	24	48	72
cornea	0	0	0	0	0	0	0	0	0	0	0	0
iris	0	0	0	0	0	0	0	0	0	0	0	0
conjunctivae
redness	1	0	0	0	1	0	0	0	1	0	0	0
chemosis	0	0	0	0	0	0	0	0	0	0	0	0
discharge	0	0	0	0	0	0	0	0	0	0	0	0

Interpretation of results:

GHS criteria not met

Conclusions:

The substance is not irritating to the eyes

Executive summary:

Three rabbits received 96 mg of the substance in the eye. Very slight irritation was seen 1 hour after installation. At the other observations (until 72 hours after instillation) no signs of irritation were found. The substance is not irritating to the eyes.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the outcome of the available studies, the substance does not need classification for skin and eye irritation according to EC No 1272/2008 (CLP Regulation)