Chromate(5-), bis[4-(hydroxy-κO)-7-[2-(2-hydroxy-1-naphthalenyl)diazenyl]-3-[2-[2-(hydroxy-κO)- 3-nitro-5-sulfophenyl]diazenyl]-2-naphthalenesulfonato(4-)]-, sodium (1:5)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

415-070-1

EC Name:

-

Molecular formula:

C52H26CrN10Na5O22S4

IUPAC Name:

chromium(3+) pentasodium bis(6-[2-(2-hydroxynaphthalen-1-yl)diazen-1-yl]-2-[2-(3-nitro-2-oxido-5-sulfonatophenyl)diazen-1-yl]-3-sulfonatonaphthalen-1-olate)

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Remarks:

(SPF-Quality)

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
-Source: Broekman Institute, Someren, The NetherTands.
-Age at study initiation: Approx. 13 weeks
-Weight at study initiation: 2451 - 2802 grams
-Housing: Individually in labelled cages with perforated floors and equipped with an automatic drinking system (ITL, Bergen, The Netherlands).
-Diet: Standard laboratory rabbit diet (LKK-20, pellet diameter 4 mm, Hope Farms,
Woerden, The Netherlands) approx. 100 g per day.
-Water: Free access to tap-water diluted with decalcified water.
-Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
-Temperature (°C): 21°C
-Humidity (%): 55 %
-Air changes (per hr): 15 air changes per hour
-Photoperiod (hrs dark / hrs light): 12 hours cycle dark/light by artificial fluorescent light

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

97 ± 1 mg per animal

Duration of treatment / exposure:

one single injection

Observation period (in vivo):

approximately 1, 24, 48 and 72 hours and 7 days after instillation of the test substance.

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

OBSERVATIONS
-Irritation:Immediately after the 24 hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.
-Viability / Mortality/ Toxicity: twice daily.
-Body Weight: once, on day of treatment (prior to instiUation).
-Staining of the treated eye was described and recorded at each observation. In case standard lighting was considered Inadequate for observing minor effects, eye examinations were performed using a pocket flash light. In cases of equivoca! results when comparing the treated and untreated eyes, the illustrated guide from the Consumer Product Safety Commission, Washington, D.C. 20207 was used for additional control purposes.

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
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:

other: all animals

Time point:

24/48/72 h

Score:

0

Results 2

Irritation parameter:

iris score

Basis:

other: all animals

Time point:

24/48/72 h

Score:

0

Results 3

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: Redness was difficult to score, due to black discolouration by the test substance.

Results 4

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Redness was difficult to score, due to black discolouration by the test substance. At 24 hours grade 2 for eyelids and at 72 hours grade 1 for sclera.

Results 5

Irritation parameter:

chemosis score

Remarks:

redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.33

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Redness was difficult to score, due to black discolouration by the test substance. At 24 grade 1 and at 72 hours grade 2 for sclera.

Results 6

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: at 24 hours grade 2 for sclera.

Results 7

Irritation parameter:

chemosis score

Basis:

animal: #2 and #3

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: at 48 and at 72 hours grade 1 for sclera.

Irritant / corrosive response data:

-Irritation: slight dulling of normal lustre was also noted on day 1 in all animals, but had disappeared after 24 hours. Treatment of the eyes with 2% fluorescein revealed no corneal epithelial damage in any of the animals, and confirmed the absence of corneal injury after 24 hours. The irritation consisted of redness (maximum grades 1 -2) and chemosis (maximum grade 2) of the conjunctivae and lacrimation. However, conjunctival redness could not be scored on day 1 and was difficult to score at the following observations, due to black discolouration by the test substance. A small amount of discharge was also noted in animals #1 and #2 during the study.
-Corrosion: there was no evidence of ocular corrosion.
-Colouration: black staining of the conjunctival tissues and of the fur on the head and paws of the animals was noted throughout the observation period.

Applicant's summary and conclusion

Interpretation of results:

other: Classified for serious eye damage (Category 1) according to the CLP Regulation (EC 1272/2008)

Conclusions:

The test substance produced irreversible effects on the rabbit eye.

Executive summary:

The eye irritation test was performed to the rabbit eye according to OECD Guidelines 405 (1987) and the method B.5 of EEC-Directive 92/69 EEC. The substance was found to cause in all animals slight dulling of normal lustre at 1 hour, disappeared after 24 hours. Treatment of the eyes with 2% fluorescein revealed no corneal epithelial damage in any of the animals, and confirmed the absence of corneal injury after 24 hours. Mean values at 24, 48 and 72 hours for iris and corneal opacity were 0 in all animals. The substance affected the conjunctivae: redness scores (mean values at 24, 48 and 72 hours ) were 0.67 for animal #1 and 1.33 for animals #2 and #3 and chemosis scores (mean values at 24, 48 and 72 hours ) were 1 for all animals. These effects on conjunctival redness and chemosis were reversible within 72 hours (animal #1) and 7 days (animals #2 and #3). However, conjunctival redness could not be scored on day of treatment and was difficult to score at all the following observations, due to black discolouration caused by the test substance. A black staining of the conjunctival tissues of all animals was noted throughout all the observation period and the reversibility could not be verified.