Tetrasodium 2,2''-diamino-8,8''-dihydroxy-4',4'''-{4-[bis(2-hydroxyethyl)amino]-1,3,5-triazine-2,6-diyldiimino}bis(naphthalene-1-azobenzene-2',6-disulfonate)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 20th to September 16th, 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Red LF 6339

IUPAC Name:

Red LF 6339

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage Scientifique des Dombes, F-01400 Chatillon sur Chalaronne / France
- Age at study initiation: 12 weeks (male), 10-11 weeks (females)
- Housing: Individually in stainless-steel cages equipped with feed hoppers and drinking water bowls. Wood bloks and haysticks were provided for gnawing.
- Diet (e.g. ad libitum): pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum
- Water (e.g. ad libitum): community tap water from Fullinsdorf ad libitum
- Acclimation: under laboratory conditions after health examination.

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

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

- Test item preparation: grounded with a pestle and mortat prior to use. 0.1 g (per animal) of test item was then weighed and applied undiluted
- Amount applied: 0.1 g

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

3 per sex

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): not rinsed after installation

SCORING SYSTEM:
Method of calculation assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69/EEC, July 31, 1992 approximately 1, 24, 48 and 72 hours as well as 7, 10, 14, 17 and 21 days after the removal of the dressing, gauze patch and test item.
When present, corrosion and/or staining of sclera and cornea by the test item were recorded and reported.

TOOL USED TO ASSESS SCORE: Varta Cliptrix dianostic lamp

OBSERVATIONS
- Viabillity/Mortality: daily from delivery of the animals to the termination of the test
- Clinical signs: daily from delivery of the animals to the termination of the test
- Body weights: at start of acclimatization, on the day of appliacation and at termination of the observation

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

iris score

Basis:

other: all animals

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 2

Irritation parameter:

chemosis score

Basis:

other: all animals

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 3

Irritation parameter:

cornea opacity score

Basis:

other: all animals

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal: 85

Time point:

24/48/72 h

Score:

0.33

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Results 5

Irritation parameter:

conjunctivae score

Basis:

animal: 86

Time point:

24/48/72 h

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 72 hrs

Results 6

Irritation parameter:

conjunctivae score

Basis:

animal: 87

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

- no clinical signs of systemic toxicity were observed
- no mortality occured
- Irritation: slight to moderate reddening of the conjuctivae was noted in all animals from 1 to 24 hours after treatment. Slight reddening continued to be observed in one animal up to the 48-hour examination and in another animal up to 72-hr examination. Slight swelling of the conjuctivae was observed in all animals 1 hour after treatment. Slight reddening of the sclerae was apparent in all animals from 1 to 24 hours after treatment and persisted in two animals up to 48-hr examination. Increased colour discharge was present in all animals 1 hour after treatment.
- Coloration: a ligtht red staining was observed in the treated eye of all animals for the majority of the observation period and was still present in one animal at termination, 21 days after treatment.
- Corrosion: no corrosion of the cornea was observed at any of the reading times.
- The mean value obtained for each of the scoring parameters indicate that the test item is not irritating

Applicant's summary and conclusion

Conclusions:

The substance is not irritant to eye, but produces red staining in the eye throughout the observation period that is still apparent in one animal at termination 21 days after treatment.