Trisodium bis[3-hydroxy-4-[(2-hydroxy-1-naphthyl)azo]naphthalene-1-sulphonato(3-)]chromate(3-)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From August 09th to September 06th, 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted 24.02.1987

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Acid Blue 193

IUPAC Name:

Acid Blue 193

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Interfauna UK Ltd Wyton, Huntingdon, England
- Age at study initiation: adult
- Weight at study initiation: the animals were weighed immediately before application of the test substance.
- Housing: the rabbits were individually housed in stainless steel cages with flat rod bases or plastica cages with perforated bases, under standardised conventional conditions.
- Diet: Standard diet "Ssniff K4" approx. 100-120g per animal/day; once a day in the morning.
- Water: tap water ad libitum.
- Acclimation period: prior to use the animals were acclimated for approx. one week in the animal room.
- Health check: approx. 24 hours before the test fur was shaved from the dorso-lateral area of the trunk (6x6 cm) of each of the rabbits. Care was taken to avoid abrading the skin. Only animals with healty and intact skin were used.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 1.5 °C
- Humidity: 60-80 %
- Air changes:12-15 times per hour
- Photoperiod: from 6 am to 6 pm, 12 hours artifical illumination.

Test system

Vehicle:

unchanged (no vehicle)

Remarks:

pulverized

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 µl of the pulverised test substance-equivalent to approx. 34 mg

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

21days

Number of animals or in vitro replicates:

3 rabbits

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: treated eye was rinsed with normale saline.
- Time after start of exposure: 24 hours after instillation.

TOOL USED TO ASSESS SCORE: optical instrument (hand slit lamp) and fluorescein. Only effects persisting for more than 24 hours were included in the evaluation.

SCORING SYSTEM
For each animal the Draize scores (cornea, iris, conjunctivae, erythema and swelling and discharge) and the Mcdonald score (aqueous humor) were used. For each animal the Draize scores recorded approx. 24, 48 and 72h after application were added up. The total of these three values was divided by 3 to give the irritation index.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

1

Reversibility:

fully reversible within: 7 days in 2/3 animals

Results 2

Irritation parameter:

iris score

Basis:

animal: 2/3

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible within: 7 days in 3/3 animals

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

2

Reversibility:

fully reversible within: 21 days in 2/3 animals

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 21 days in 3/3 animals

Irritant / corrosive response data:

Exposure of the eye to test substance caused reactions of the mucous membranes and effects of the cornea in all three animals. Additionally, in all three animals the iris was transiently affected and discharge occured. In one animal signs was fully reversible within 21 days, in the second animal corneal signs did not prove to be completely reversible within 21 days and in the third animal a slight erythema was observed on day 21. This evidence indicates that the product may be considered as "severely irritating to the eye".

Any other information on results incl. tables

Individual reactions

Animal	Reactions	Score at							Mean 24/48/72 hrs
		1 hr	24 hrs	48 hrs	72 hrs	7 d	14 d	21 d
D11	Corneal opacity	1	1	1	1	0	0	0	1.00
D7	Corneal opacity	1	1	1	1	1*	1+	1+	1.00
E8	Corneal opacity	1	1	1*	1*	0	0	0	1.00
D11	Iris	0	0	0	1	0	0	0	0.33
D7	Iris	-	1	1	1	0	1	0	1.00
E8	Iris	0	1	0	1	0	0	0	0.67
D11	Conjunctival redness	e	2k	2k	2k	0	1	1	2.00
D7	Conjunctival redness	e	2	2k	2k	1	1	0	2.00
E8	Conjunctival redness	e	2	2k	2k	0	0	0	2.00
D11	Conjunctival chemosis	2v	2	1	1	0	1	0	1.33
D7	Conjunctival chemosis	2v	2	1	1	1	0	0	1.33
E8	Conjunctival chemosis	2v	2	1	1	0	0	0	1.33

e = evaluation not possible due to colouration by test substance

k = conjunctivae and nictitating membrane: injection of blood vessels

v = conjunctivae and nictitating membrane: strong formation of vesicles

* = lower margin of cornea: vascularisation

+ = lower and upper margin of cornea: vascularisation

Applicant's summary and conclusion

Interpretation of results:

other: Eye Dam 1 (H318), according to the CLP Regulation (EC) No 1272/2008

Remarks:

Migrated information

Conclusions:

Eye Dam 1, H318

Executive summary:

The irritant/corrosive potential of test substance was studied on the eye of rabbit in accordance with the OECD guideline 405. After gently pulling the lower lid away from the eyeball a volume of 100 µl of the pulverised test substance-equivalent to approx. 34 mg- was placed into the conjunctival sac of one eye of each of three rabbits. 24 hours after instillation of test substance, the treated eye was rinsed with normale saline.

Exposure of the eye to test substance caused reactions on mucous membranes and effects on cornea in all three animals. Additionally in all three animals the iris was transiently affected and discharge occured. In one animal signs was fully reversible within 21 days, in the second animal corneal signs did not prove to be completely reversible within 21 days and in the third animal a slight erythema was observed on day 21. This evidence indicates that the product may be considered as "severely irritating to the eye".

Conclusion

The mean values from gradings at 24, 48 and 72 hours recorded in the study were equal to 1 for corneal opacity in all animals, lower than 1 for iritis in 2 out of three animals, equal to 2 for conjunctival redness and lower than 2 for oedema in all animals. Corneal opacity was not fully recovered within 21 days in one animals, as well as in another animal the conjunctival redness was not recovered withing 21 days.

Due to the fact that at least in one animal effects on the cornea have not fully reversed within an observation period of normally 21 days, a classification as capable to causes serious eye damage (i.e. category 1, H318), according to the CLP Regulation (EC) No 1272/2008, is applied.