Benzenesulfonic acid, 4-chloro-2-[2-[4,5-dihydro-3-methyl-5-oxo-1-(3-sulfophenyl)-1H-pyrazol-4-yl]diazenyl]-5-methyl-, ammonium salt (1:2)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Details on test material:

- Appearance: Yellow powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Lot/batch number of test material: KL384
- Expiration date of the lot/batch: not specified
- Purity test date: not specified


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in the dark
- Stability under storage conditions: not specified
- Stability under test conditions: not specified
- Solubility and stability of the test substance in the solvent/dispersant/vehicle/test medium: not applicable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: no

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Froxfield (U.K.) Ltd., Petersfield, Hampshire, England or Charles River (UK) Ltd., Margate, Kent, England
- Age at study initiation: approximately 12 to 19 weeks
- Weight at study initiation: 2.7 to 3.9 kg
- Housing: one animal per cage
- Diet: standard laboratory diet SDS Stanrab (P) Rabbit Diet ad libitum
- Water: drinking water ad libitum
- Acclimation period: yes; duration not specified
- Microbiological status when known: not specified
- Method of randomisation in assigning animals to test and control groups: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): approximately 19°C
- Humidity (%): 30 - 70
- Air changes (per hr): approximately 19
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: To: not specified

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): approx. 76 mg (occupying a volume of 0.1 ml)

Duration of treatment / exposure:

single application

Observation period (in vivo):

7d

Number of animals or in vitro replicates:

3 females

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no

SCORING SYSTEM:
Examination of the eyes was made after 1 hour and 1, 2, 3 (equivalent to 24, 48 and 72 hours after instillation), 4 and 7 days after instillation.

Ocular irritation was assessed using the prescribed numerical system:
- 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 areas, details of iris slightly obscured - 2
Nacreous areas, 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 the most severe reading of palpebral and bulbar conjunctivae, as compared to the control eye)
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 (lids and/or nictating membranes)
No swelling - 0
Any swelling above normal (includes nictating 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

Any other lesion not covered by this scoring system, was described.

TOOL USED TO ASSESS SCORE: handheld light

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

mean of all 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 2

Irritation parameter:

iris score

Basis:

mean

Remarks:

mean of all 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

mean of all 3 animals

Time point:

24/48/72 h

Score:

0.333

Max. score:

3

Reversibility:

fully reversible within:

Remarks:

2 d

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

mean of all 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 5

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 1 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within:

Remarks:

24 h

Results 6

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 1 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within:

Remarks:

24 h

Results 7

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: 1 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within:

Remarks:

24 h

Irritant / corrosive response data:

Dulling of the normal lustre of the cornea was seen in two animals one hour after instillation only.
No other corneal damage or iridial inflammation was observed.
Considerable swelling with partial eversion of the eyelids was seen in one animal one hour after instillation only. Temporary mild conjunctival reactions were seen in the remaining animals.
The eyes were normal two days after instillation

Other effects:

There were no signs of toxicity or ill health in any rabbit during the observation period.

Any other information on results incl. tables

Tab. 1: Ocular reactions observed after instillation of the test substance

Rabbit number	Region of eye		One hour	Day after instillation
				I23 47
543	Cornea		D	000 00
	Iris		0	00 0 0 0
	Conjunctiva	Redness	I	I0 0 0 0
		Chemosis	I	00000
580	Cornea		D	0 0 0 0 0
	Iris		0	0 000 0
	Conjunctiva	Redness	I	I000 0
		Chemosis	2	0 0 0 0 0
582	Cornea		0	0 0 0 0 0
	Iris		0	0 0 0 0 0
	Conjunctiva	Redness	I	1000 0
		Chemosis	1	00 0 00

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Instillation of the test substance into the rabbit eye elicited dulling of the cornea and mild transient conjunctival irritation. All reaction has resolved within 2d. The substance is therefore considered to be non-irritating.

Executive summary:

A study was performed to assess the eye irritation potential of the test substance to the rabbit.

The method followed was that described in EEC Methods for the determination of toxicity, Annex to Directive 92/69/EEC (OJ No. L383A, 29.12.92), Part B, Mediod B.5. Acute toxicity (eye irritation). The study was conducted under GLP conditions.

Three rabbits were each administered a single ocular dose of 76 mg of the test substance and observed for seven days after instillation. A single instillation of the test material into the eye of the rabbit elicited dulling of the cornea and transient mild conjunctival irritation. All reactions had resolved 2 days after instillation.