Chromate(2-), [2,4-dihydro-4-[(2-hydroxy-4-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)][3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulfonato(3-)]-, disodium

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

no data

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:

1978

Report date:

1978

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

(observation period was up to 7 d instead of 21 d; eyes were not examined at 1 h after test substance application; clinical observations for systemic effects was not done)

Principles of method if other than guideline:

not applicable

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Chromate(2-), [2,4-dihydro-4-[(2-hydroxy-4-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)][3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulfonato(3-)]-, disodium

EC Number:

274-386-6

EC Name:

Chromate(2-), [2,4-dihydro-4-[(2-hydroxy-4-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)][3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulfonato(3-)]-, disodium

Cas Number:

70209-87-9

Molecular formula:

C32H21CrN10Na2O11S

IUPAC Name:

Disodium-[2,4-dihydro-4-[(2-hydroxy-4-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)][3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulfonato(3-)]-chromate(2-)

Test material form:

not specified

Details on test material:

- Name of test material (as cited in study report): FAT 20060/B
- Substance type: Dye
- Analytical purity: Approx. 82%
- Lot/batch No.: EN 78708

Test animals / tissue source

Species:

rabbit

Strain:

Himalayan

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 1.5 to 2 kg
- Housing: The rabbits were caged singly in metal cages
- Diet: Standard rabbit food - NAFAG, Gossau SG (ad libitum)
- Water: ad libitum
- Acclimation period: 4 d

ENVIRONMENTAL CONDITIONS
- Temperature: 20±1 °C.
- Humidity: 55±5 %
- Photoperiod: 10 h (light cycle day)

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: the second eye on the treated animals served as control

Amount / concentration applied:

- Amount applied: 100 mg
- Concentration: Undiluted

Duration of treatment / exposure:

Unwashed eye: Duration of treatment same as observation period
Washed eye: 30 seconds

Observation period (in vivo):

The eyes were examined for any ocular reaction approximately 1, 2, 3, 4 and 7 d (i.e., 24, 48, 72, 96 and 168 h) after the treatment

Number of animals or in vitro replicates:

6 (i.e., 3 males and 3 females)

Details on study design:

PROCEDURE:
-The test was carried out in accordance with the procedure set out in the Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics (1959) of the US Association of Food and Drug Officials (AFDO) which is similar to OECD Guideline 405.
-Only rabbits with normal ophthalmic findings were used for the test.
-The test substance in an amount of 100 mg was inserted into the conjunctival sac of the left eye of the rabbits and the eyelids were gently closed for one second. The right eye was not treated and served as an untreated control.
-In 3 of the 6 rabbits approximately 30 seconds after treatment the treated eye was flushed with 10 mL of lukewarm water.
-The eye irritation was evaluated with a slit-lamp and was scored for each rabbit on 1, 2, 3, 4 and 7 d.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Results 2

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

13.9

Max. score:

80

Reversibility:

fully reversible

Remarks on result:

other: Individual scores were not cited.

Results 3

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

10

Reversibility:

fully reversible

Remarks on result:

other: Individual scores were not cited.

Results 4

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

14.4

Max. score:

20

Reversibility:

fully reversible

Remarks on result:

other: Individual scores were not cited.

Irritant / corrosive response data:

The irritation index was calculated to be 13.9 for the cornea, 0 for the iris and 14.4 for the conjunctivae. The test substance caused moderate overall eye irritation to rabbits. Rinsing the eyes following instillation increased the effect of the test substance.

Other effects:

none

Any other information on results incl. tables

Results of the eye irritation:

Unwashed eyes after application of test substance:

Days after Appl.	Rabbit 1 (M)			Rabbit 2 (M)			Rabbit 3 (M)
	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva
1	15	0	18	10	0	18	10	0	16
2	15	0	16	10	0	18	10	0	14
3	15	0	16	10	0	18	10	0	14
4	15	0	16	10	0	18	10	0	10
7	10	0	10	10	0	14	0	0	6

Cornea: 10.6; Iris: 0; Conjunctivae: 14.8

 

Eyes washed after application of test substance:

Days after Appl.	Rabbit 4 (F)			Rabbit 5 (F)			Rabbit 6 (F)
	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva
1	10	0	18	20	0	18	15	0	20
2	10	0	16	40	0	18	15	0	20
3	10	0	14	40	0	16	15	0	18
4	5	0	10	40	0	14	15	0	18
7	0	0	0	20	0	14	5	0	6

 

Cornea: 17.3; Iris: 0; Conjunctivae: 14

 

R (effect of rinsing) = unrinsed eyes/rinsed eyes (i.e., 8.4/10.4) = 0.8

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

Under the conditions of the study, the test substance was considered to be moderately irritating to the eye.

Executive summary:

An in vivo study was conducted to evaluate the acute eye irritation potential of the test substance (at ca. 82 % purity) in rabbits according to the procedure described in the Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics (1959) of the US Association of Food and Drug Officials (AFDO), which is similar to OECD Guideline 405 but with few deviations. 100 mg of the test substance was placed into the conjunctival sac of the left eye of six rabbits (i.e., 3 male and 3 females). The right eye remained untreated and was used as control. In three of the six rabbits, washing was done with 10 mL of lukewarm water after 30 seconds of test substance instillation. Assessment of ocular damage/irritation was made with a slit-lamp and scored for each individual rabbit approximately on 1, 2, 3, 4 and 7 d following treatment. The total maximum score was 80 for the cornea, 10 for the iris, 20 for the conjunctivae and 110 for the entire eye. The irritation index was calculated to be 13.9, 0 and 14.4 for the cornea, iris and conjunctivae, respectively. Rinsing the eyes following instillation increased the effect of the test substance. Under the conditions of the study, the test substance was considered to be moderately irritating to the eye.