Sodium bis[2-chloro-4-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-5-hydroxybenzenesulphonamidato(2-)]chromate(1-)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October 1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

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: "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US "Hazardous Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965).

Deviations:

not specified

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Sodium bis[2-chloro-4-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-5-hydroxybenzenesulphonamidato(2-)]chromate(1-)

EC Number:

266-577-8

EC Name:

Sodium bis[2-chloro-4-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-5-hydroxybenzenesulphonamidato(2-)]chromate(1-)

Cas Number:

67109-27-7

Molecular formula:

C32H24Cl2CrN10O8S2.Na

IUPAC Name:

sodium bis[2-chloro-4-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-5-hydroxybenzenesulphonamidato(2-)]chromate(1-)

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Remarks:

Porton strain

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Inhouse breeding
- Age at study initiation: 12 - 16 weeks
- Weight at study initiation: Males 2-93 kg and Female 2.78 kg
- Housing: caged singly
- Diet: A commercial irradiated diet (Styles-Oxoid) was fed ad libitum
- Water: Sterile filtered water ad libitum
- Acclimation period: One week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 1
- Humidity (%): 50 - 70
- Photoperiod (hrs dark / hrs light): artificial light for 10 h

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g of the test substance.

Duration of treatment / exposure:

single application

Observation period (in vivo):

1 and 24 h and after 2, 3, 6, 10 and 14 days.

Number of animals or in vitro replicates:

6 rabbits (3 males and 3 females)

Details on study design:

The eyes of the experimental animals were examined and found normal prior to the test. 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. The right eye served as a control. After 30 seconds the compound was, as far as possible, flushed out of the eyes of three of the rabbits with approximately 200 ml of warm water.

SCORING SYSTEM: Evaluation of the Eye Reactions.
CORNEA:
A. Opacity-degree of density (area most dense taken for reading)
-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 Total maximum = 80

IRIS
A. Values
-Normal: 0
-Folds above normal, congestion, swelling, circum-corneal 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

CONJUNCTIVAE
A. Redness (refers to palpebral 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 canthus 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 Total maximum =20

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks on result:

other: Eyes rinsed and eyes not rinsed

Results 2

Irritation parameter:

cornea opacity score

Remarks:

Area involved

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

3.55

Max. score:

4

Reversibility:

not fully reversible within: 14 d

Remarks on result:

other:

Remarks:

Eyes rinsed and eyes not rinsed

Results 3

Irritation parameter:

iris score

Basis:

mean

Remarks:

Animal no 11, 12, 13, 15 and 16

Time point:

24 h

Score:

1

Max. score:

2

Reversibility:

not fully reversible within: 14 d

Results 4

Irritation parameter:

iris score

Remarks:

Eyes not rinsed

Basis:

animal #4

Remarks:

Rabbit 14

Time point:

24 h

Score:

0

Max. score:

2

Results 5

Irritation parameter:

iris score

Remarks:

Eyes rinsed

Basis:

mean

Remarks:

Rabbit 11,12,13

Time point:

48 h

Score:

1

Max. score:

2

Reversibility:

not fully reversible within: 14 d

Results 6

Irritation parameter:

iris score

Remarks:

Eyes not rinsed

Basis:

mean

Remarks:

Rabbit 14,15,16

Time point:

48 h

Score:

0

Max. score:

2

Results 7

Irritation parameter:

iris score

Remarks:

Eyes rinsed

Basis:

mean

Remarks:

Rabbit 11,12,13

Time point:

72 h

Score:

1

Max. score:

2

Reversibility:

not fully reversible within: 14 d

Results 8

Irritation parameter:

iris score

Remarks:

Eyes not rinsed

Basis:

mean

Remarks:

Rabbit 14,15,16

Time point:

72 h

Score:

0

Max. score:

2

Results 9

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

2.11

Max. score:

3

Reversibility:

not fully reversible within: 14 d

Results 10

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

1.83

Max. score:

4

Reversibility:

not fully reversible within: 14 d

Results 11

Irritation parameter:

overall irritation score

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

4.44

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

The primary eye irritation score is 4.44.

Any other information on results incl. tables

Conjunctiva:

On initial application of the compound there appeared to be a moderately painful reaction. A mild to moderate conjunctival reaction was seen in all treated eyes one hour after the application of the compound. At 24 h there was very little change from the initial reaction but thereafter these started to improve until at termination of study on day 14 one washed eye was normal, 5/5 unwashed and 2/5 washed eyes were showing a very slight to slight reaction and the remaining unwashed eye a moderate reaction.

Cornea:

Slight opacity with damage to the surface epithelium was seen in all treated eyes at one hour. This was still present at 24 hours and did not vary much for up to 72 h. At 6 days the 3 washed eyes were normal but the unwashed eyes were all showing slight pannus formation. In 2 of these the pannus increased in severity slightly over the remainder of the test period and was still present at termination.

Iris:

Increased folding was seen in 3/5 unwashed and 2/5 washed eyes at 24 h. The washed eyes were normal by 48 h and one unwashed eye at 14 days. The other 2 unwashed eyes were still showing increased folding at termination. All eyes would have eventually returned to normal if sufficient time had been allowed to elapse. However, 2 out of the 3 unwashed eyes might have had some very small residual scarring of the cornea. Generally, the reactions seen in washed eyes, particularly after the initial reaction, were less severe and recovery was faster than those in unwashed eyes.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

FAT 20028/B is considered as an irritant to the rabbit eye with the primary eye irritation index is 4.44.

Executive summary:

The study was carried out with FAT 20028/B in NZW rabbit according to the procedure adopted was the technique described in the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US "Hazardous Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965). The substance was tested on 6 rabbits (3 males/3 females), New Zealand White rabbit (Porton strain). Only rabbits showing normal ophthalmic findings were included in the test. 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. The right eye served as a control. After 30 seconds the compound was, as far as possible, flushed out of the eyes of three of the rabbits with approximately 200 ml. of warm water. 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.

 

RESULTS:

Conjunctiva:

On initial application of the compound there appeared to be a moderately painful reaction. A mild to moderate conjunctival reaction was seen in all treated eyes one hour after the application of the compound. At 24 h there was very little change from the initial reaction but thereafter these started to improve until at termination of study on day 14 one washed eye was normal, 5/5 unwashed and 2/5 washed eyes were showing a very slight to slight reaction and the remaining unwashed eye a moderate reaction.

 

Cornea:

Slight opacity with damage to the surface epithelium was seen in all treated eyes at one hour. This was still present at 24 hours and did not vary much for up to 72 h. At 6 days the 3 washed eyes were normal but the unwashed eyes were all showing slight pannus formation. In 2 of these the pannus increased in severity slightly over the remainder of the test period and was still present at termination.

 

Iris:

Increased folding was seen in 3/5 unwashed and 2/5 washed eyes at 24 h. The washed eyes were normal by 48 h and one unwashed eye at 14 days. The other 2 unwashed eyes were still showing increased folding at termination. All eyes would have eventually returned to normal if sufficient time had been allowed to elapse. However, 2 out of the 3 unwashed eyes might have had some very small residual scarring of the cornea. Generally, the reactions seen in washed eyes, particularly after the initial reaction, were less severe and recovery was faster than those in unwashed eyes. Therefore, FAT 20028/B is to be considered as a minimal irritant to the eye of rabbits and the primary eye irritation index is 4.44.