Reaction products of diazotised 3-amino-2-hydroxy-5-nitrobenzenesulphonic acid and 3-amino-4-hydroxy-5-nitrobenzenesulphonic acid, coupled with resorcinol, metalized with Ferric Chloride, condensed with Formaldehyde, sodium salts

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:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

study performed on the analogue substance; the read across justification is detailed in section 13. The Reliability of the Source Study is 2.

Justification for type of information:

Justification for read-across is detailed at section 13.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: Real Decreto 363/1995 of 10 March

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Real Decreto 1078/93 of 2 July

Deviations:

not specified

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Similar Substance 01

IUPAC Name:

Similar Substance 01

Test material form:

solid: particulate/powder

Test animals / tissue source

Species:

rabbit

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Housing: Tecniplast inox cages.
- Diet: 150 g per day standard diet.
- Water: tap water; ad libitum.
- Acclimation period: 7 days.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 2°C
- Humidity: 55 ± 25 %
- Air changes: 15 ACH
- Photoperiod: 12 hours dark

Test system

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

0.1 g fine powder.

Observation period (in vivo):

at 1, 24, 48 and 72 hours and at 7, 14 and 21 day after application.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no

SCORING SYSTEM:
Cornea
Opacity: degree of density
No ulceration or opacity........................................................................................... 0
Scattered or diffuse areas of opacity .................................................................... 1
Easily discernible translucent area, details of iris slightly obscured........................2
Nacrous area, 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, congestión, swelling ................................................... 1
No reaction to light, hemorrhage, gross destructlon (any or all of these) ...............2

Conjunctivae - redness
Blood vessels normal ...............................................................................................0
Some blood vessels definitely hyperemic (injected) ................................................1
Diffuse, crimson color, individual vessels not easily discernible............................. 2
Diffuse beefy red......................................................................................................3

Conjunctivae - chemosis
No swelling............................................................................................................... 0
Any swelling above normal (includes nictitating membranes)..................................1
Obvious swelling with partial eversión of lids.......................................................... 2
Swelling with lids about half closed .........................................................................3
Swelling with lids more than half-closed...................................................................4

TOOL USED TO ASSESS SCORE: hand-slit lamp.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

other: 3/3 animals

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible

Results 2

Irritation parameter:

iris score

Basis:

other: 3/3 animals

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible

Results 3

Irritation parameter:

conjunctivae score

Basis:

other: 2/3 animals

Time point:

24/48/72 h

Score:

>= 2

Reversibility:

not fully reversible within: 21 days

Remarks:

At rabbit #2 and #3, on 21th day, was given a conjunctival redness score of 1.

Results 4

Irritation parameter:

chemosis score

Basis:

other: 2/3 animals

Time point:

24/48/72 h

Score:

>= 2

Reversibility:

fully reversible

Any other information on results incl. tables

Mean values for cornea, iris, conjunctiva at 24, 48 and 72 hours.

	Rabbit #1	Rabbit #2	Rabbit #3	Mean
Corneal opacity	1	1	1	1
Iritis	0	0.67	1	0.55
Conjunctival redness	2	2.67	3	2.55
Conjunctival oedema	0.67	3	3.67	2.44

	Individual findings - Rabbit #1
	1 h	24 hrs	48 hrs	72 hrs	7 days	14 days	21 days
Corneal opacity	-	1	1	1	0	0	0
Iritis	1	0	0	0	0	0	0
Conjunctival redness	3	2	2	2	2	1	0
Conjunctival oedema	2	1	1	0	0	0	0

	Individual findings - Rabbit #2
	1 h	24 hrs	48 hrs	72 hrs	7 days	14 days	21 days
Corneal opacity	0	1	1	1	0	0	0
Iritis	0	1	1	0	0	0	0
Conjunctival redness	0	3	3	2	2	2	1
Conjunctival oedema	3	4	3	2	1	0	0

	Individual findings - Rabbit #2
	1 h	24 hrs	48 hrs	72 hrs	7 days	14 days	21 days
Corneal opacity	0	1	1	1	1	0	0
Iritis	0	1	1	1	1	0	0
Conjunctival redness	0	3	3	3	2	2	1
Conjunctival oedema	3	4	4	3	2	1	0

Applicant's summary and conclusion

Interpretation of results:

other: Classified as Eye Irrit. 2 according to the CLP Regulation (EC 1272/2008).

Conclusions:

The substance is classified as Eye Irrit. 2 under the Regulation EC 1272/2008 (CLP).

Executive summary:

The test was performed according to the Real Decreto 363/1995.

The eye irritation potential was investigated by instillation of the equivalent of 0.1 g/animal of the test substance. The scores of each animal at the reading times of 24, 48 and 72 hours was used in calculating the respective mean values for each type of lesion, as stated by the Regulation EC 1272/2008 (CLP).

Scores for corneal opacity and iritis were lower than the threshold value of 1 for 3/3 rabbits.

Scores for conjunctival redness and oedema were higher than the threshold value of 2 in 2/3 rabbits.

Every type of lesion was fully reversible within 21 days except the conjunctival redness response, which was identified in 2/3 rabbits on the 21th day of the test (graded as 1 according to the used scoring system).

From the present results the substance should be classified as corrosive to the eyes because the produced effects were not fully reversible within 21 days. However, the following considerations should be underlined.

Since the study did foresee the rising of eyes after the application of the test item in its solid form, the results of the test can be considered as an overestimation of the true irritating effects strictly related to the substance chemical/toxicological properties. Indeed, due to the intrinsic irritating nature of dusty materials, this overestimation is true especially for the conjunctival redness response and the fact that this lesion was not fully reversible within 21 days can be attributed to the lack of eye rinsing. Moreover, from the individual scoring of each animal, it can be observed a progressive reduction of this lesion, suggesting that the effect will most likely not be persistent.

For the above mentioned reason, the substance does not meet the criteria for classification as Eye Damage 1. Instead, a classification as Eye Irrit 2, under the Regulation EC 1272/2008 (CLP), is warranted.