Trisodium bis[3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-4-hydroxy-5-nitrobenzenesulphonato(3-)]chromate(3-)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

other: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Test procedures cannot be subsumed under testing guideline today commonly used in the ambit of the REACH Regulation, nevertheless they followed other internationally accepted guideline, are well documented and scientifically acceptable. Justification for Read Across is detailed in the endpoint summary.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1978

Report date:

1978

Materials and methods

Principles of method if other than guideline:

The procedure used was the test for eye irritants described in the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959), the US Association of Food and Drug Officials (AFDO)..

GLP compliance:

no

Remarks:

pre GLP

Test material

Test material information

Constituent 1

Reference substance name:

Similar Substance 01 (AR357)

IUPAC Name:

Similar Substance 01 (AR357)

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: animals were housed individually in metal cages.
- Diet: ad libitum, standard rabbit food - NAFAG, Gossau SG.
- Water: ad libitum.
- Acclimation period: prior to treatment they were adapted to our laboratories for a minimum of 4 days. Only rabbits with normal ophthalmic findings were used for these tests.

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

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 g
- Application: the test material in an amount of 0.1 g was inserted into the conjunctival sac of the left eye of the rabbits and the lids were gently closed for one second.

Duration of treatment / exposure:

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

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3 males and 3 females

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: in 3 of the 6 rabbits the treated eye was flushed with 10 ml of lukewarm water.
- Time after start of exposure: approximately 30 seconds after treatment.

TOOL USED TO ASSESS SCORE: hand-slit lamp

SCORING SYSTEM
The eye irritation was appraised on day 1, 2, 3, 4 and 7 and was scored for each individual rabbit.
The primary eye irritation index represents the sum of the mean values for cornea, iris and conjunctivae. This procedure was carried out separately for unrinsed and rinsed eyes respectively.

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, haemorrhage, 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 = 2

The total maximum score was thus 80 for the cornea, 10 for the iris, 20 for the conjunctivae and 110 for the entire eye. The severity of overall eye irritation
was rated as:
0 - 10 minimal
11 - 25 slight
26 - 56 moderate
57 - 84 marked
above 84 extreme

The ratio unrinsed/rinsed eyes indicates the effect of the rinsing the eyes 30 seconds after instillation of the compound.
< 1 = rinsing increased the effect of the compound
1.1 - 1.5 = practically no effect
1.6 - 2 = little but assessable effect
2.1 - 4 = good effect
4.1 - 10 = very good effect
> 10 = action of the compound practically abolished

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Score:

13.9

Max. score:

80

Remarks on result:

other: moderate individual scores were not cited

Results 2

Irritation parameter:

iris score

Basis:

mean

Score:

0

Max. score:

10

Remarks on result:

other: moderate individual scores were not cited

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Score:

14.4

Max. score:

20

Remarks on result:

other: moderate 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.

Any other information on results incl. tables

Results of the eye irritation

 

Unwashed eyes after application of test substance

Days after application	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 application	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:

other: not applicable, according to the CLP Regulation

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

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

Executive summary:

Method

The substance was tested on 6 rabbits; amounts of 0.1 g of the test substance were introduced into the conjunctival sac. The eyes of 3 animals were rinsed 30 seconds after application. The eye irritation was appraised with a slit lamp on day 1, 2, 3, 4 and 7.

Result

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.

Conclusion

The criteria for the evaluation of the eye irritation potential followed in the test are different from those reported into the OECD guideline. The scoring system used for the cornea opacity and degree of density, for the iris and conjunctivae redness and chemosis is the same of that reported into the OECD, but the timing for the removal of the substance are very different (30 seconds in the current study and 1 hours in the OECD).

Unfortunately, the raw tables are not included into the study report and thus, the results cannot be completely re-evaluated, according to the CLP Regulation (EC 1272/2008).

On the basis of the calculation system for the scores, it can only be deduced that iris was not impaired in both the cases of rinsed and not rinsed eyes. Cornea and conjunctivae were affected, but the injury extension cannot be evaluated.

In conclusion, the test item effects recorded in the current study cannot be assessed according to the CLP Regulation (EC 1272/2008).