Reaction products of 6-amino-4-hydroxynaphthalene-2-sulphonic acid and 7-amino-4-hydroxynaphthalene-2-sulphonic acid, condensed with Formaldehyde and Sodium hydrogensulfite, coupled with diazotised Sodium 2-amino-4,6-dinitrophenoxide, metalized with Sodium Dichromate, sodium salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

1981

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of 6-amino-4-hydroxynaphthalene-2-sulphonic acid and 7-amino-4-hydroxynaphthalene-2-sulphonic acid, condensed with Formaldehyde and Sodium hydrogensulfite, coupled with diazotised Sodium 2-amino-4,6-dinitrophenoxide, metalized with Sodium Dichromate, sodium salts

EC Number:

947-452-3

Molecular formula:

not applicable

IUPAC Name:

Reaction products of 6-amino-4-hydroxynaphthalene-2-sulphonic acid and 7-amino-4-hydroxynaphthalene-2-sulphonic acid, condensed with Formaldehyde and Sodium hydrogensulfite, coupled with diazotised Sodium 2-amino-4,6-dinitrophenoxide, metalized with Sodium Dichromate, sodium salts

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: abbaye de Bellefontaine (49122 Bégrolles en Mauges, France).
-Weight at study initiation: 2,588 kg.
-Housing: the rabbits were individually housed in polystyrene cages :
0,35 x 0,55 x 0,32 m. Each cage was equipped with a food container and a water bottle.
-Diet: the animals were fed ad libitum with a certified pelleted diet Rabbit Diet SOC (Special Diets Services Ltd, witham, Essex, England) during the study.
-Water: the animals had free access to _tap water filtered with Millipore filters in water bottles.
-Acclimation period: 7 days. During this period, the animals were observed daily.

ENVIRONMENTAL CONDITIONS
-Temperature (°C): 18°C ± 3 °C
-Humidity (%): 50 ± 20 %.
-Air changes (per hr): The incoming, non-recycled air, was filtered by ani absolute filter.
-Photoperiod (hrs dark / hrs light): The light/dark cycle was 12 hours per day.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 g

Duration of treatment / exposure:

Single injection

Observation period (in vivo):

5 days

Number of animals or in vitro replicates:

6 rabbits

Details on study design:

SCORING SYSTEM
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.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3, #6

Time point:

24/48/72 h

Score:

0.67

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: unrinsed eye

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #4

Time point:

24/48/72 h

Score:

1

Reversibility:

fully reversible within: 5 days

Remarks on result:

other: unrinsed eye

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: unrinsed eye

Results 4

Irritation parameter:

iris score

Basis:

animal: #1, #2, #5

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: unrinsed eye

Results 5

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: unrinsed eye

Results 6

Irritation parameter:

iris score

Basis:

animal: #4, #6

Time point:

24/48/72 h

Score:

0.67

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: unrinsed eye

Results 7

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal: #1, #6

Time point:

24/48/72 h

Score:

2

Reversibility:

fully reversible within: 5 d

Remarks on result:

other: unrinsed eye

Results 8

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal: #2, #5

Time point:

24/48/72 h

Score:

1

Reversibility:

fully reversible within: 5 d

Remarks on result:

other: unrinsed eye

Results 9

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal: #3, #4

Time point:

24/48/72 h

Score:

1.33

Reversibility:

fully reversible within: 5 d

Remarks on result:

other: unrinsed eye

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.33

Reversibility:

fully reversible within: 5 d

Remarks on result:

other: unrinsed eye

Results 11

Irritation parameter:

chemosis score

Basis:

animal: #2, #4, #5, #6

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: unrinsed eye

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: unrinsed eye

Irritant / corrosive response data:

One hour after instillation of the test sample a swelling with lids about half closed occurred.
Redness of the conjunctivae and iridial and corneal reactions could not be recorded due to the staining of the eye by the compound.
On Day 1, the swelling disappeared in four animals and decreased in the two others. Some blood vessels were hyperaemic in two animals and a diffuse crimson colour was observed in four animals.
A moderate circumcorneal hyperaemia of the iris was observed in three animals.
A diffuse area of opacity covered an area between 1/4 and 1/2 of the cornea in four animals and less than 1/4 in one animal.
On Day 2, chemosis disappeared in five animals, redness decreased in two anhnals. The iridial reaction disappeared in one animal and the surface area of opacity decreased in three animals.
On Day 3, the iridial reactions disappeared in all animals and the corneal reactions in four animals. The ocular reactions were similar on Day 4 and had disappeared on Day 5.

Applicant's summary and conclusion

Interpretation of results:

other: not classified according to the CLP classification (EC 1272/2008)

Conclusions:

The substance was found to be not irritating for rabbit eye according to the CLP classification (EC 1272/2008).

Executive summary:

An acute eye irritation study was performed with the test sample after instillation into the conjunctival sac of six New-Zealand rabbits according to the OECD Guideline 405 (1981).

The scores obtained related to the effects observed into the treated eye were used to calculate the mean scores 24/48/72 h. Corneal opacity were 0.67 in four animals (#1, #2, #3 and #6), 1 in animal #4 and 0 in animal #5. Iris scores were 0 in three animals (#1, #2 and #5), 0.33 in animal #3 and 0.67 in two animals (#4 and #6). Conjunctival erythema (redness) scores were 2 in two animals (#1 and #5), 1 in two animals (#2 and #5) and 1.33 in two animals (#3 and #4). Conjunctival oedema (chemosis) were 0 in four animals (#2, #4, #5 and #6), 1.33 in animal #1 and 0.33 in animal #3. All the effects observed were fully reversible within 5-day observation period after the test item application.