reaction mass of: pentasodium bis[6-anilino-3,5'-disulfonatonaphthalene-2-azobenzene-1,2'-diolato]cobaltate(III);tetrasodium [6-anilino-3,5'-disulfonatonaphthalene-2-azobenzene-1,2'-diolato][6-anilino-5'-sulfamoyl-3-sulfonatonaphthalene-2-azobenzene-1,2'-diolato]cobaltate(III);trisodium bis[6-anilino-5'-sulfamoyl-3-sulfonatonaphthalene-2-azobenzene-1,2'-diolato]cobaltate(III)

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:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2002

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

444-290-0

EC Name:

-

Cas Number:

508202-43-5

Molecular formula:

Not applicable

IUPAC Name:

tricobalt(3+) dodecasodium tris(2-[2-(2-oxido-5-sulfamoylphenyl)diazen-1-yl]-6-(phenylamino)-3-sulfonatonaphthalen-1-olate) tris(2-[2-(2-oxido-5-sulfonatophenyl)diazen-1-yl]-6-(phenylamino)-3-sulfonatonaphthalen-1-olate)

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
-Source: Elevage Scientifique des Dombes F-01400 Chatillon sur Chalaronne / France
-Age at study initiation:
♂ 10 weeks
♀ 11 weeks
-Weight at study initiation:
♂ 2001 g
♀ 2133-2091 g
-Housing: individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks and haysticks 4646 were provided for gnawing.
-Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum
-Water: Community tap water from Füllinsdorf, ad libitum.

ENVIRONMENTAL CONDITIONS
-Temperature: 17-23 °C
-Humidity: 30-70 %
-Air changes: 10-15 air changas per hour
-Photoperiod: 12 hours cycle dark/light
-Other: music was played during the daytime light period.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount: 0.1 g

Duration of treatment / exposure:

Single injection

Observation period (in vivo):

1, 24, 48 and 72 hours, as well as 7,10,14,17 and 21 days after application.

Number of animals or in vitro replicates:

3 ( 1 ♂ and 2 ♀)

Details on study design:

The treated eyes were not rinsed after installation.

TOOL USED TO ASSESS SCORE
-Eye examinations were made with a Varta Cliptrix diagnostic-lamp

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 and #2

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: not rinsed

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: not rinsed

Results 3

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: not rinsed

Results 4

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal: #1 and #3

Time point:

24/48/72 h

Score:

1.33

Reversibility:

fully reversible within: 10 days

Remarks on result:

other: not rinsed

Results 5

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: not rinsed

Results 6

Irritation parameter:

chemosis score

Basis:

animal: #1 and #2

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: not rinsed

Results 7

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48 hours

Remarks on result:

other: not rinsed

Irritant / corrosive response data:

IRRITATION
Corneal opacity due to staining produced by the test item was observed in one animal from 1 to 72 hours after treatment. No abnormal findings were observed in the iris of any animal at any of the measurement intervals.
Reddening of the conjunctivae could not be assessed in one animal at the 1-hour reading due to the staining produced by the test item. Slight to moderate reddening was apparent in all animals from 1 to 24 hours after treatment. Slight reddening was still present in all animals at 48- and 72-hours reading and continued to be observed in one animal up to 7 days and in another animal up to 10 days.
Slight to moderate swelling was noted in all animals at the 1-hour examination. The severity of swelling gradually decreased and was no longer evident in any animal 48 hours after treatment.
Assessment of the sclerae was not possible in one animal at the 1-hour reading due to the staining produced by the test item. Moderate reddening was noted in the other two animals.
Slight to moderate reddening was still present in all animals at the 24-hour reading and continued to be observed in one animal up to 48 hours. Ocular discharge with moistening of the lids and hairs just adjacent to the lids was apparent in all animals at the 1 -hour examination. No abnormal findings were observed in the treated eye of any animal 21 days after treatment, the end of the observation period for all animals, except violet staining in one animal.

CORROSION
No corrosion of the cornea was observed at any of the reading times.

Other effects:

COLORATION
Color remnants of the test item were observed in the eye or conjunctival sac of all animals 1 hour after treatment. Marked violet staining caused by the test item was evident in all animals at the 1-hour reading. Light to marked violet staining was then observed in all animals up to 7 days. Light violet staining persisted in one animal up to termination.

VIABILITY/MORTALITY AND CLINICAL SIGNS
No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred

Any other information on results incl. tables

 

.

Applicant's summary and conclusion

Interpretation of results:

other: Classified according to the CLP Regulation (EC n. 1272/2008) Eye Dam. 1

Conclusions:

The test item induced irreversible damage to the rabbit eye, as the violet coloured staining persisted in one animal throughout the observation period.

Executive summary:

The eye irritation test was performed to the rabbit eye according to OECD Guidelines 405 (2002) and the method B.5 of EEC-Directive 92/69 EEC. 0.1 g of test item were installed into the left eye of 3 young adult New Zealand White rabbits. Scoring of irritation effects was performed after test item application at 1, 24, 48 and 72 hour, as well as 7, 10, 14, 17 and 21 day.

Mean values at 24, 48 and 72 hours for iris were 0.00 in all animals. The other mean values for corneal opacity were 0.00, 0.00 and 1.00; for the conjunctivae were 1.33, 1.00 and 1.33 (redness) and 0.00, 0.00 and 0.33 (chemosis), respectively in animals #1, #2 and #3. The instillation into the eye resulted in mild, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae, discharge and chemosis. These effects were reversible and were no longer evident 21 days after treatment, the end of the observation period for all animals.

No corrosion was observed at any of the measuring intervals.

A slight opacity of the cornea was noted in one animal due to the staining produced by the test item. Marked violet staining caused by the test item was evident in all animals at the 1-hour reading. Light to marked violet staining was then observed in all animals up to 7 days. Light violet staining persisted in one animal up to termination.

Thus, the test item induced irreversible damage to the rabbit eye: the violet coloured staining which persisted in one animal throughout the observation period.