Sodium 4(-2-hydroxy-1-naphthylazo)naphthalenesulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 16th Feb. 2004 to 8th Mar. 2004

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

24 April 2002

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

31 July 1992

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Sodium 4(-2-hydroxy-1-naphthylazo)naphthalenesulphonate

EC Number:

216-760-3

EC Name:

Sodium 4(-2-hydroxy-1-naphthylazo)naphthalenesulphonate

Cas Number:

1658-56-6

Molecular formula:

C20H13N2NaO4S

IUPAC Name:

NA

Test material form:

solid: particulate/powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Granja Cunicola San Bernardo, S.L. (Tulebras, Navarra, Spain).
- Age at study initiation: 9-10 weeks.
- Weight at study initiation: 2.6 - 3.1 kg.
- Housing: individually in stainless steel cages (52 x 58 x 43 cm) with a grille floor and placed on a rack.
- Diet: free access throughout the Study to a standard diet for rabbits, Panlab 112 (Panlab, S.L., Barcelona, Spain analysed by the manufacturer.
- Water: water ad libitum provided in plastic bottles.
- Acclimation period: 7 days.

ENVIRONMENTAL CONDITIONS
- Temperature: 19-23 “C
- Humidity: 30-60 %
- Photoperiod: 12 hours of light (7:00 am to 7:00 pm) and 12 hours of darkness every 24 hours.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 g

Duration of treatment / exposure:

ca 1 hours

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3 rabbits

Details on study design:

ADMINISTRATION
The test item was administered initially to one animal and afterwards applied to two more animals.

REMOVAL OF TEST SUBSTANCE
- Washing: with physiological saline before the first observation.

SCORING SYSTEM
The behaviour of animals was observed immediately following installation of the product and they were then returned to thei cages and observed at regular intervals so as not to miss any serious effect of the test item.
The degree of ocular irritation was assessed after approximately 1, 24, 48, 72 hour(s) and 7 days after the treatment, following the criteria given below.


CORNEA
- Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity: 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible: 1
Easily discernible translucent area, details of iris slightly obscured: 2
Nacreous areas, no details of iris visible, size of pupil barely discernible: 3
Opaque cornea, iris not discernible through the opacity: 4

- Area of cornea affected
No ulceration or opacity: 0
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

IRIS
Normal: 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection, any of these or any combination thereof, iris still reacting to light (sluggish reaction is positive): 1
No reaction to light, haemorrhage, gross destruction (any or all of these): 2

CONJUNCTIVAE
- Redness: (refers to palpebral and bulbar conjunctivae, excluding cornea and iris).
Blood vessels normal 0
Some blood vessels definitely hyperaemic (injected): 1
Diffuse, crimson colour, individual vessels not easily discernible: 2
Diffuse beefy red: 3

- Chemosis: lids and/or nictating membrane
No swelling: 0
Any swelling above normal (includes nictating membrana): 1
Obvious swelling with partial eversion of lids: 2
Swelling with lids about half closed: 3
Swelling with lids more than half closed: 4

- 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, in a considerable area around the eye: 3.

TOOL USED TO ASSESS SCORE:
All the observations except the one at one hour after administration, the observation of the cornea was completed with the instillation of an aqueous solution of 2% sodium fluorescein (COLIRCUST FLUORESCEINA®, Laboratorios Cusi, S.A., Batches S03-D and 3CUJ2E). This procedure involved the instillation of the aqueous solution of fluorescein in the test area, followed by washing the area with a 0.9% physiological saline solution. Once the excess fluorescein had been removed, the corneal alterations were observed with the aid ofa transilluminator with a cobalt blue filter.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #3

Time point:

24/48/72 h

Score:

0

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.67

Reversibility:

fully reversible within: 72 hours

Results 3

Irritation parameter:

iris score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0

Results 4

Irritation parameter:

chemosis score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48 hours

Results 5

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Reversibility:

fully reversible within: 72 hours

Results 6

Irritation parameter:

conjunctivae score

Basis:

animal: #2 and #3

Time point:

24/48/72 h

Score:

1.33

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

- Initally treated animal: one hour after the application, redness diffuse, crimson colour, individual vessels not easily discernible (grade 2) was recorded, accompanied by obvious swelling with partial eversion of lids (grade 2). In this animal lacrimation with moistening of the lids and surrounding fur (grade 2) was also observed.
The lesions in the conjunctivae were difficult to assess one hour after the administration, because of the stain caused by the test item. Twenty-four hours after the administration, this animal presented some blood vessels definitely hyperaemic (grade l), with slight swelling (grade 1). Forty-eight hours after the administration, the blood vessels definitely hyperaemic (grade 1) persisted, but the swelling had remitted completely.
No ocular lesions were recorded in this animal 72 hours and 7 days after the administration.

- The two more animals: one hour after the administration, two animals were seen to have redness diffuse, crimson colour, individual vessels not easily discernible (grade 2), with obvious swelling with partial eversion of lids (grade 2) in one of them, and slight edema (grade 1) in the other one.
Both animals presented lacrimation with moistening of the lids and surrounding fur (grade 2). The lesions in the conjunctivae were difficult to assess in both animals one hour after the administration because of the stain caused by the test item. Twenty-four and 48 hours after the administration, scattered or diffuse areas of opacity, details of iris clearly visible (grade 1) was recorded, covering a quarter of the area or less, but not zero (grade 1).
Twenty-four hours after the administration, both animals were seen to have redness diffuse, crimson colour, individual vessels not easily discernible (grade 2), accompanied by slight swelling (grade 1). Forty-eight and 72 hours after the administration, some blood vessels definitely hyperaemic (grade 1) were observed. No ocular lesions were recorded in these animals 7 days after the administration.

Any other information on results incl. tables

Animal No	Reaction	1 h	24 hrs	48 hrs	72 hrs	7 days	Mean 24/48/72 hrs
#1	Cornea opacity	0	0	0	0	0	0.00
	Iris	0	0	0	0	0	0.00
	Conjunctival redness	2	1	1	0	0	0.67
	Conjunctival chemosis	2	1	0	0	0	0.33
#2	Cornea opacity	0	1	1	0	0	0.67
	Iris	0	0	0	0	0	0.00
	Conjunctival redness	2	2	1	1	0	1.33
	Conjunctival chemosis	2	1	0	0	0	0.33
#3	Cornea opacity	0	0	0	0	0	0.00
	Iris	0	0	0	0	0	0.00
	Conjunctival redness	2	2	1	1	0	1.33
	Conjunctival chemosis	1	1	0	0	0	0.33

Applicant's summary and conclusion

Interpretation of results:

other: not classified according to the CLP Regulation (EC) No 1272/2008

Conclusions:

The test item was found not to be irritanting to the rabbit eyes.

Executive summary:

The degree of primary eye irritation caused by the test substance was evaluated in a study performed according to the OECD Guideline No. 405 (2002) and the Method B.5 of the European Communities Commission Directive (92/69/EEC) of 31 July 1992d. 0.1 g of test item was administered to the right eye of three animals; the left eye remained untreated and was used as a control. The observations following the administration were at 1, 24, 48, 72 hour and 7, 14 day.

The mean values (24/48/72 h) for the iris were 0 in all animals, while for the corneal opacity were 0 in two animals (#1 and #3) and 0.67 in one animal (#2). The conjunctival oedema (chemosis) mean scores were 0.33 in all animals, while the conjunctival redness ones were 0.67 in one animal (#1) and 1.33 in the other two animals (#2 and #3). No ocular lesions were recorded in any of the treated animals 7 days after the administration.