Disodium 2,5-dichloro-4-[4-[[5-[[(dodecyloxy)carbonyl]amino]-2-sulphonatophenyl]azo]-4,5-dihydro-3-methyl-5-oxo-1H-pyrazol-1-yl]benzenesulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November from 05th to 19th, 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

treated eye not rinsed

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

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

Test material

Test material information

Constituent 1

Reference substance name:

Acid Yellow 218

IUPAC Name:

Acid Yellow 218

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: David Percival Ltd., Moston, Sandbach, Cheshire, U.K.
- Age at study initiation: approximately twelve to sixteen weeks old.
- Weight at study initiation: 2.72 - 2.86 kg
- Housing: individually housed in suspended metal cages.
- Diet: free access to food (Rabbit Diet, Preston Farmers Limited, Nerv Leake, Boston, Lincolnshire, U.K.l) was allowed throughout the study.
- Water: free access to mains drinking water.
- Acclimation period: minimum acclimatisation period of five days.
- Health check: immediately before the start of the test, both eyes of the three provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. Animals showing evidence of ocular Iesions were rejected and replaced.

ENVIRONMENTAL CONDITIONS
- Temperature: 16 - 24 °C
- Relative humidity: 51 - 72 %
- Air changes: approximately 15 changes per hour.
- Photoperiod: 12 hours light and 12 hours darkness.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 ml (approx. 96 mg)

Observation period (in vivo):

14 days

Number of animals or in vitro replicates:

One rabbit was initially treated; after consideration of the ocular responses produced in the first treated animal, two additional animals were treated.
1 male and 2 females

Details on study design:

TREATED EYES
In order to minimise pain on instillation of the test material, one drop of local anaesthetic "Ophthaine", 0.5 % proxymetacaine hydrochloride was instilled into both eyes of these animals 1-2 minutes before treatment.

TOOL USED TO ASSESS SCORE: examination of the eye was facilitated by use of the light source from a standard ophthalmoscope.

SCORING SYSTEM
Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation described in Draize J.H. 1959, Association of Food and Drug Officials of the united States, Austin, Texas, "The Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics"). Any other adverse ocular effects were also noted.
Additional observations were made on days seven and fourteen to assess the reversibility of the ocular effects.

CORNEA:
A Opacity and degree of density (most dense area scored) .
No opacity: 0
Scattered or diffuse area, details of iris clearly visible: 1
Easily discernible translucent areas, details of iris slightly obscured: 2
Opalescent areas, no details of iris visible, size of pupil barely discernible: 3
Opaque, iris invisible: 4

B Area of cornea involved
One quarter (or less) but not zero: 1
Greater than one quarter, but less than half: 2
Greater than half, but less than three quarters: 3
Greater than three quarters, up to whole area: 4
A × B × 5; Maximum possible score = 80

IRITIS:
A Values
Normal: 0
Folds above normal, congestion, swelling, circumcornea injection (any or all of these or combination of any thereof) iris still reacting to light (sluggish reaction is positive): 1
No reaction to light, hemorrhage, gross destruction (any or all of these): 2
A × 5; Maximum possible score = 10

CONJUNCTIVAE:
A Redness (refers to lids and bulbar conjunctivae excluding cornea and iris)
Vessels normal: 0
Vessels definitely injected above normal: 1
More diffuse, deeper crimson red, individual vessels not easily discernible: 2
Diffuse beefy red: 3

B Chemosis
No swelling: 0
Any swelling above normal (includes nictitating membrane): 1
Obvious swelling with partial eversion of lids: 2
Swelling with lids about half closed: 3
Swelling with lids about half closed to completely closed: 4

C Discharge
No discharge: 0
Any amount different from normal (does not include small amounts observed in inner cantus of normal animals): 1
Discharge with moistening of the lids and hairs just adjacent to lids: 2
Discharge with moistening of the lids and hairs, and considerable area around the eye: 3
(A + B + C) × 2; Maximum possible score = 20

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: 14 days

Results 2

Irritation parameter:

iris score

Basis:

animal: 2/3

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible within: 72 hours

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 2/3

Time point:

24/48/72 h

Score:

>= 2

Reversibility:

fully reversible within: 14 days

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 2/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Vocalisation was noted in one animal approximately ten seconds after dosing.
Faint yellow-coloured staining caused by the test material was noted in all treated eyes during the study. This did not prevent accurate evaluation of ocular reactions.
A dulling of the normal lustre of the corneal surface was noted in two treated eyes one hour after treatment. Areas of diffuse corneal opacity were noted in one treated eye one hour after treatment and in all treated eyes at the 24-hour observation. Diffuse corneal opacity persisted in all treated eyes up to the 72-hour observation and in one treated eye at the 7-day observation.
Iridial inflammation was noted in all treated eyes one hour after treatment, at the 24 and 48-hour observations and in one treated eye at the 72-hour observation.
Moderate to severe conjunctival irritation was noted in all treated eyes one hour after treatment with moderate conjunctival irritation at the 24 and 48-hour observations. Minimal to moderate conjunctival irritation was noted in all treated eyes at the 72-hour observation with minimal conjunctival irritation in one treated eye at the 7-day observation.
Residual test material was noted around the treated eyes of all animals during the study.
All treated eyes appeared normal 7 - 14 days after treatment.
The test material produced a maximum group mean score of 36.0 and was classified as a moderate irritant to the rabbit eye according to a modified Kay and Calandra classification system.

Any other information on results incl. tables

Animal	Reaction	24 hrs	48 hrs	72 hrs	7 days	14 days	Mean 24/48/72 hrs
70 male	Corneal opacity	1	1	1	1	0	1.00
88 female	Corneal opacity	1	1	1	0	0	1.00
92 female	Corneal opacity	1	1	1	0	0	1.00
70 male	Iris	1	1	1	0	0	1.00
88 female	Iris	1	1	0	0	0	0.67
92 female	Iris	1	1	0	0	0	0.67
70 male	Conjunctival redness	2	2	2	1	0	2.00
88 female	Conjunctival redness	2	2	1	0	0	1.70
92 female	Conjunctival redness	3	3	1	0	0	2.33
70 male	Conjunctival chemosis	3	2	2	0	0	2.33
88 female	Conjunctival chemosis	2	1	0	0	0	1.00
92 female	Conjunctival chemosis	2	2	1	0	0	1.67

Individual reactions

	70 male						88 female					92 female
	1 hr	24 hrs	48 hrs	72 hrs	7 days	14 days	1 hr	24 hrs	48 hrs	72 hrs	7 days	1 hr	24 hrs	48 hrs	72 hrs	7 days
Cornea opacity	1	1	1	1	1	0	d	1	1	1	0	d	1	1	1	0
Area	4	4	2	2	1	0	4	3	3	2	0	4	4	4	2	0
Score	20	20	10	10	5	0	0	15	15	10	0	0	20	20	10	0
Iris	1	1	1	1	0	0	1	1	1	0	0	1	1	1	0	0
Score	5	5	5	5	0	0	5	5	5	0	0	5	5	5	0	0
Conjuntival redness	2s	2	2	2	1	0	2s	2	2	1	0	2s	3	3	1	0
Conjuntival chemosis	3	3	2	2	0	0	2	2	1	0	0	3	2	2	1	0
Conjuntival discharge	3re	2re	1re	0re	0	0	3re	1re	1re	0	0	3re	2re	1re	0re	0
Score	16	14	10	10	2	0	14	10	8	2	0	16	14	12	5	0
Total score	41	39	25	25	7	0	19	30	28	12	0	21	39	37	14	0

d = dulling of the nornal lustre of the corneal surface

re = resídual test material around the eye

s = faint yellow staining

Applicant's summary and conclusion

Interpretation of results:

other: Eye Irrit. 2 (H319), according to the CLP Regulation (EC 1272/2008)

Conclusions:

Eye irritant.

Executive summary:

The study was performed to assess the irritancy potential of the test material following a single application to the rabbit eye. The method used followed the recommendations of OECD guideline 405.

One rabbit was initially treated. A volume of 0.1 ml of test material, which was found to weigh approximately 96 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye. The left eye remained untreated and was used for control purposes. Immediately after administration of test substance, an assessment of the initial local pain reaction was made. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

Vocalisation was noted in one animal approximately ten seconds after dosing. Faint yellow-coloured staining caused by the test material was noted in all treated eyes during the study. This did not prevent accurate evaluation of ocular reactions. A dulling of the normal lustre of the corneal surface was noted in two treated eyes one hour after treatment. Areas of diffuse corneal opacity were noted in one treated eye one hour after treatment and in all treated eyes at the 24-hour observation. Diffuse corneal opacity persisted in all treated eyes up to the 72-hour observation and in one treated eye at the 7-day observation. Iridial inflammation was noted in all treated eyes one hour after treatment, at the 24 and 48-hour observations and in one treated eye at the 72-hour observation. Moderate to severe conjunctival irritation was noted in all treated eyes one hour after treatment with moderate conjunctival irritation at the 24 and 48-hour observations. Minimal to moderate conjunctival irritation was noted in all treated eyes at the 72-hour observation with minimal conjunctival irritation in one treated eye at the 7-day observation. All treated eyes appeared normal 7 - 14 days after treatment.

Conclusion

The mean values from gradings at 24, 48 and 72 hours were equal to 1 for corneal opacity in all the animals tested, lower than 1 for iritis in 2 out of 3 animals, equal/higher than 2 for conjunctival redness in 2 out of 3 animals and lower than 2 for conjunctival oedema in 2 out of 3 animals. All treated eyes appeared normal 7 - 14 days after treatment.