Disodium 2,5-dichloro-4-(5-hydroxy-3-methyl-4-(sulphophenylazo)pyrazol-1-yl)benzenesulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1976

Report date:

1976

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: "Hazardous-Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12

Version / remarks:

February 1965

Deviations:

yes

Remarks:

slight modification

GLP compliance:

no

Remarks:

pre GLP

Test material

Test material information

Constituent 1

Reference substance name:

Acid Yellow 017

IUPAC Name:

Acid Yellow 017

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age at study initiation: 12-16 weeks.
- Weight at study initiation: average body weights of 2.80 kg males and 2.60 kg females.
- Housing: rabbits were caged singly.
- Diet: commercial irradiated diet (Styles-Oxoid), ad libitum.
- Water: sterile filtered water, ad libitum.
- Acclimation period: one week prior to the start of the trial.
- Health check: the eyes of the experimental animals were examined and found normal prior to the test.

ENVIRONMENTAL CONDITIONS
- Temperature: 20 ± 1 °C
- Relative humidity: 50-70 %
- Photoperiod: artificial light for 10 hours daily from 03.00 - 18.00 hours.

Test system

Controls:

yes

Amount / concentration applied:

100 mg of the test compound were instilled into the conjunctival sac of the left eye. The eyelids were then held closed for 1 second.

Duration of treatment / exposure:

30 seconds in three animals; in the remaining animals eyes were not washed.

Number of animals or in vitro replicates:

3 males and 3 females

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: with approximately 200 ml of warm water.
- Time after start of exposure: 30 seconds.

TOOL USED TO ASSESS SCORE
Corneal damage was assessed after staining with fluorescein.

SCORING SYSTEM
The rabbits were examined 1, 24, 48 and 72 hours after "application of the test compound and for any further period that was considered necessary. The ocular reactions were scored by "the method described in "Appraisal of the Safety of Chemicals in Food Drugs and Cosmetics", published by the Association of Food and Drug Officials of the U.S.A.

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 x B x 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 x 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) x 2; Maximum possible score = 20

The eye reactions were assessed as follows:
0 Non irritant
0 - 10 Minimally irritant
11 - 25 Slightly irritant
26 - 56 Moderately irritant
57 - 84 Markedly irritant
> 84 Extremely irritant

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: 6/6, both rinsed and not rinsed eyes

Time point:

24/48/72 h

Score:

< 1

Results 2

Irritation parameter:

iris score

Basis:

animal: 6/6, both rinsed and not rinsed eyes

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible within: 72 hrs

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3, rinsed eyes

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 6 days

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3, not rinsed eyes

Time point:

24/48/72 h

Score:

2

Reversibility:

fully reversible within: 10 days

Results 5

Irritation parameter:

chemosis score

Basis:

animal: 6/6, both rinsed and not rinsed eyes

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 10 days

Irritant / corrosive response data:

Minimal irritant (score 0 - 10)
A slight to mild conjunctival reaction was seen in all treated eyes one hour after application of the compound. Generally this had improved very slightly by 24 hours and thereafter continued to do so until all eyes were normal by 6 days (3/3 washed eyes) and 10 days (3/3 unwashed eyes).
Slight opacity of the cornea with damage to the surface epithelium was seen in one unwashed eye at one hour only Increased folding of the iris was noted in one washed eye at 24 and 48 hours.
There was very little difference in the reactions seen between washed and unwashed eyes, apart from the washed eyes recovering slightly sooner.

Any other information on results incl. tables

Overview of results - Rinsed eyes

	24 hrs	48 hrs	72 hrs	Mean 24/48/72 hrs
Cornea
Animal 14	0	0	0	0.00
Animal 15	0	0	0	0.00
Animal 16	0	0	0	0.00
Iris
Animal 14	1	1	0	0.66
Animal 15	0	0	0	0.00
Animal 16	0	0	0	0.00
Conjunctival redness
Animal 14	2	2	1	1.66
Animal 15	2	2	1	1.66
Animal 16	2	1	1	1.33
Conjunctival chemosis
Animal 14	0	0	0	0.00
Animal 15	1	0	0	0.33
Animal 16	0	0	0	0.00

Overview of results - Not rinsed eyes

	24 hrs	48 hrs	72 hrs	Mean 24/48/72 hrs
Cornea
Animal 11	0	0	0	0.00
Animal 12	0	0	0	0.00
Animal 13	0	0	0	0.00
Iris
Animal 11	0	0	0	0.00
Animal 12	0	0	0	0.00
Animal 13	0	0	0	0.00
Conjunctival redness
Animal 11	2	2	2	2.00
Animal 12	2	2	2	2.00
Animal 13	2	2	2	2.00
Conjunctival chemosis
Animal 11	1	1	1	1.00
Animal 12	1	1	1	1.00
Animal 13	1	1	0	0.66

Details on results - Rinsed eyes

Time score	1 hr			24 hrs			48 hrs			72 hrs			6 days			10 days
Animal N.	14	15	16	14	15	16	14	15	16	14	15	16	14	15	16	14	15	16
Cornea
A Opacity
B Area involved
a: AxBx5 (max 80)
Iris				1			1
b: score x 5 (max 10)				5			5
Conjunctivae
A Redness	1	1	1	2	2	2	2	2	1	1	1	1
B Chemosis	1	1	1		1
C Discharge	2	1	1
c: (A+B+C) x 2 (Max 20)	8	6	6	4	6	4	4	4	2	2	2	2
Total = a+b+c (Max: 110)	8	6	6	4	6	4	4	4	2	2	2	2

Not rinsed eyes

Time score	1 hr			24 hrs			48 hrs			72 hrs			6 days			10 days
Animal N.	11	12	13	11	12	13	11	12	13	11	12	13	11	12	13	11	12	13
Cornea
A Opacity		1
B Area involved		1
a: AxBx5 (max 80)		5
Iris
b: score x 5 (max 10)
Conjunctivae
A Redness	2	2	2	2	2	2	2	2	2	2	2	2	1	1	1
B Chemosis	1	2	2	1	1	1	1	1	1	1		1	1
C Discharge	1	1	1
c: (A+B+C) x 2 (Max 20)	5	10	10	6	6	6	6	6	6	6	4	6	4	2	2
Total = a+b+c (Max: 110)	8	15	10	6	6	6	6	6	6	6	4	6	4	2	2

Applicant's summary and conclusion

Conclusions:

Minimal irritant

Executive summary:

Method

The eye irritation was determined in an irritation test on rabbits according to the" Hazardous-Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965), similar to OECD guideline 405.

Result

Minimal irritant