1,3'-Bipyridinium, 5'-[[4-[[4-[(1',2'-dihydro-6'-hydroxy-3,4'-dimethyl-2'-oxo[1,3'-bipyridinium]-5'-yl)azo]benzoyl]amino]phenyl]azo]-1',2'-dihydro-6'-hydroxy-3,4'-dimethyl-2'-oxo-, salt with 2-hydroxypropanoic acid (1:2)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Remarks:

source of read across

Adequacy of study:

key study

Study period:

From 29th February to 4th March, 1988

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:

1988

Report date:

1988

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

1981

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Similar substance 01

IUPAC Name:

Similar substance 01

Specific details on test material used for the study:

For the purpose of the study the test material was ground to a fine powder using a mortar and pestle before use.

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: at the start of the study the animals were approximately twelve to sixteen weeks old.
- Weight at study initiation: at the start of the study the animals weighed 2.72 - 2.96 kg.
- Housing: individually housed in suspended metal cages.
- Diet: Rabbit Diet, Preston Farmers Limited, New Leake, Boston, Lincolnshire, U.K., ad libitum.
- Water: drinking water, ad libitum.
- Acclimation period: minimum acclimatisation period of five days.

ENVIRONMENTAL CONDITIONS
- Temperature: 17 - 19 °C
- Humidity: 40 - 48 %
- Air changes: approximately 15 changes per hour.
- Photoperiod: the lighting was controlled by a time switch to give 12 hours light and 12 hours darkness.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

A volume of 0.1 ml of the test material (as measured by gently compacting the required volume into an adapted syringe) which was found to weigh approximately 64 mg was placed into the right eye of each rabbit by gently pulling the lower lid away from the eyeball to form a cup into which the test material was dropped. The upper and lower eyelids were held together for about one second immediately after application, to prevent loss of the test material from the eye, and then released.

Observation period (in vivo):

72-hour

Number of animals or in vitro replicates:

Three rabbits

Details on study design:

Immediately before commencement of the test, both eyes of each rabbit provisionally selected, were examined for evidence of ocular irritation or defect using the light source from an ophthalmoscope. Animals showing evidence of ocular lesions were rejected and replaced.

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

SCORING SYSTEM
Assessment of damage/irritation was made 1, 24, 48 and 72 hours following treatment, according to the numerical evaluation given (i.e. 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").

Grades of ocular lesions
CORNEA
Opacity: Degree of density (area most dense taken for reading).
No ulceration or opacity 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre) details of iris clearly visible 1
Easily discernible translucent area, details of iris slightly obscured 2
Opalescent area, no details of iris visible, size of pupil barely discernible 3
Opaque cornea, iris not discernible through the opacity 4

IRIS
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive) 1
No reaction to light, haemorrhage, gross destruction (any or all of these) 2

CONJUNCTIVAE
Redness: (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Blood vessels normal 0
Some blood vessels definitely hyperaemic (injected) 1
Diffuse, crimson colour, individual vessels not easily discernible 2
Diffuse beefy red 3

CHEMOSIS Value
Chemosis: Lids and/or nictitating membranes.
No swelling 0
Any swelling above normal (includes nictitating membranes) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids more than half closed 4

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

Results 2

Irritation parameter:

iris score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible within: 24 hours

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 48 hours

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 48 hours

Irritant / corrosive response data:

No ocular irritation was noted at the 72-hour observation and the study was terminated.

A dulling of the normal lustre of the corneal surface and iridial inflammation were noted in all treated eyes one hour after treatment. No adverse corneal or
iridial effects were noted at subsequent observations.
Moderate conjunctival irritation was also noted in all treated eyes one hour after instillation. The conjunctival irritation regressed and all treated eyes appeared normal at the 48-hour observation.

Any other information on results incl. tables

Individual and mean scores for cornea, iris and conjuncivae.

Rabbit	Time after treatment (h)	Corneal opacity	Iridial inflammation	Conjunctival redness	Conjunctival chemosis
80	1	D	1	2	2
	24	0	0	1	0
	48	0	0	0	0
	72	0	0	0	0
Mean 24, 48 and 72 hrs		0.0	0.0	0.3	0.0
41	1	D	1	2	2
	24	0	0	1	1
	48	0	0	0	0
	72	0	0	0	0
Mean 24, 48 and 72 hrs		0.0	0.0	0.3	0.3
34	1	D	1	2	2
	24	0	0	1	1
	48	0	0	0	0
	72	0	0	0	0
Mean 24, 48 and 72 hrs		0.0	0.0	0.3	0.3

D: dulling of corneal surface

Applicant's summary and conclusion

Interpretation of results:

other: not classified, according to the CLP Regulation (EC 1272/2008)

Conclusions:

Not eye irritant.

Executive summary:

A study was performed to assess the irritation of the test material to the eye of the New Zealand White rabbit. The method followed OECD guideline 405. A volume of 0.1 ml of the test material was placed into the right eye of each rabbit. The upper and lower eyelids were held together for about one second immediately after application, to prevent loss of the test material from the eye, and then released. Assessment of damage/irritation was made 1, 24, 48 and 72 hours following treatment. No ocular irritation was noted at the 72-hour observation and the study was terminated. A dulling of the normal lustre of the corneal surface and iridial inflammation were noted in all treated eyes one hour after treatment. No adverse corneal or iridial effects were noted at subsequent observations. Moderate conjunctival irritation was also noted in all treated eyes one hour after instillation. The conjunctival irritation regressed and all treated eyes appeared normal at the 48-hour observation.

Conclusion

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for iritis, lower than 2 for both conjunctival redness and oedema, in all the three tested animals.

Therefore, the substance does not meet the criteria to be classified as eye irritating, according to the CLP Regulation (EC 1272/2008).