A 2:1:1 mixture of: trisodium N(1')-N(2):N(1''')-N(2'')-η-6-[2-amino-4-(or 6)-hydroxy-(or 4-amino-2-hydroxy)phenylazo]-6''-(1-carbaniloyl-2-hydroxyprop-1-enylazo)-5',5'''-disulfamoyl-3,3''-disulfonatobis(naphthalene-2,1'-azobenzene-1,2'-diolato-O(1),O(2'))-chromate; trisodium N(1')-N(2):N(1''')-N(2'')-η-6,6''-bis(1-carbaniloyl-2-hydroxyprop-1-enylazo)-5',5'''-disulfamoyl-3,3''-disulfonatobis(naphthalene-2,1'azobenzene-1,2'-diolato-O(1),O(2'))-chromate; trisodium N(1')-N(2):N(1''')-N(2'')-η-6,6''-bis[2-amino-4-(or 6)-hydroxy-(or 4-amino-2-hydroxy)phenylazo]5',5'''-disulfamoyl-3,3''-disulfonatobis(naphthalene-2,1'azobenzene-1,2'-diolato-O(1),O(2'))-chromate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 29. Sep. to 13. Oct. 1987

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

1987

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

1984

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

A 2:1:1 mixture of: trisodium N(1')-N(2):N(1''')-N(2'')-η-6-[2-amino-4-(or 6)-hydroxy-(or 4-amino-2-hydroxy)phenylazo]-6''-(1-carbaniloyl-2-hydroxyprop-1-enylazo)-5',5'''-disulfamoyl-3,3''-disulfonatobis(naphthalene-2,1'-azobenzene-1,2'-diolato-O(1),O(2'))-chromate; trisodium N(1')-N(2):N(1''')-N(2'')-η-6,6''-bis(1-carbaniloyl-2-hydroxyprop-1-enylazo)-5',5'''-disulfamoyl-3,3''-disulfonatobis(naphthalene-2,1'azobenzene-1,2'-diolato-O(1),O(2'))-chromate; trisodium N(1')-N(2):N(1''')-N(2'')-η-6,6''-bis[2-amino-4-(or 6)-hydroxy-(or 4-amino-2-hydroxy)phenylazo]5',5'''-disulfamoyl-3,3''-disulfonatobis(naphthalene-2,1'azobenzene-1,2'-diolato-O(1),O(2'))-chromate

EC Number:

402-850-1

EC Name:

A 2:1:1 mixture of: trisodium N(1')-N(2):N(1''')-N(2'')-η-6-[2-amino-4-(or 6)-hydroxy-(or 4-amino-2-hydroxy)phenylazo]-6''-(1-carbaniloyl-2-hydroxyprop-1-enylazo)-5',5'''-disulfamoyl-3,3''-disulfonatobis(naphthalene-2,1'-azobenzene-1,2'-diolato-O(1),O(2'))-chromate; trisodium N(1')-N(2):N(1''')-N(2'')-η-6,6''-bis(1-carbaniloyl-2-hydroxyprop-1-enylazo)-5',5'''-disulfamoyl-3,3''-disulfonatobis(naphthalene-2,1'azobenzene-1,2'-diolato-O(1),O(2'))-chromate; trisodium N(1')-N(2):N(1''')-N(2'')-η-6,6''-bis[2-amino-4-(or 6)-hydroxy-(or 4-amino-2-hydroxy)phenylazo]5',5'''-disulfamoyl-3,3''-disulfonatobis(naphthalene-2,1'azobenzene-1,2'-diolato-O(1),O(2'))-chromate

Molecular formula:

not applicable for UVCB substance

IUPAC Name:

trichromium(3+) nonasodium 6-[2-(2-amino-4-hydroxyphenyl)diazen-1-yl]-2-[2-(2-oxido-5-sulfamoylphenyl)diazen-1-yl]-3-sulfonatonaphthalen-1-olate 6-[2-(2-amino-6-hydroxyphenyl)diazen-1-yl]-2-[2-(2-oxido-5-sulfamoylphenyl)diazen-1-yl]-3-sulfonatonaphthalen-1-olate 6-[2-(4-amino-2-hydroxyphenyl)diazen-1-yl]-2-[2-(2-oxido-5-sulfamoylphenyl)diazen-1-yl]-3-sulfonatonaphthalen-1-olate tris(6-{2-[(1Z)-2-hydroxy-1-(phenylcarbamoyl)prop-1-en-1-yl]diazen-1-yl}-2-[2-(2-oxido-5-sulfamoylphenyl)diazen-1-yl]-3-sulfonatonaphthalen-1-olate)

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: Kleintierfarm Madörin AG, 4414 Füllinsdorf, CH
- Age at study initiation: 14 to 15 weeks
- Weight at study initiation: 2.5 to 2.6 kg
- Housing: individually in stainless steel cages equipped with an automatic cleaning and drinking system (Dipl. Ing. W. Ehret Gmbh, Versuchstiertechnik, 7830 Emmendingen, Germany)
- Diet: ad libitum; pelleted standard Kliba 341, batch 30/87 rabbit maintenance diet ("Kliba", Klingentalmühle AG, 4303 Kaiseraugst, CH) (analytical test report demonstrates suitability)
- Water: ad libitum; community tap water from Itingen (bacteriological assay and chemical water analysis demonstrated suitability)
- Acclimation period: 4 days under laboratory conditions after veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature: 20 °C ± 3 degrees
- Humidity: 40 to 70 %
- Air changes: 10 to 15 per hour
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount: 0.1 g per animal
- Concentration: undiluted; solid

Duration of treatment / exposure:

One single application

Observation period (in vivo):

14 days

Number of animals or in vitro replicates:

Males: 1
Females: 2
Total: 3

Details on study design:

TREATMENT
- The test item was placed in the conjunctival sacs of the left eye of each animal after gently pulling the lower lid away from the eye ball. The lids were then gently held together for about 1 second to prevent loss of the test item.

REMOVAL OF TEST SUBSTANCE :
- Washing: no washing

TOOL USED TO ASSESS SCORE:
- Eye examinations were made with a slit-lamp 30 SL (C. Zelss AG, Zürich, CH) and a "Varta Cliptrix" diagnostic-lamp (A. Riegger, Basel, CH)

SCORING SYSTEM:
- Irritation scores of each animal at 1, 24, 48 and 72 hours and 7 and 14 days were measured.
- The corrosive properties of the test item and the colour of the treated eye were described and recorded.
- The following numerical system was used to grade the eye lesions:

CORNEAL IRRITATION: opacity; degree of density (densest area used for assessment)
0 No ulceration or opacity
1* Scattered or diffuse areas of opacity (other than slight dulling of normal luster), details of iris clearly visible
2* Easily discernible translucent area, details of iris slightly obscured
3* Nacrous area, no details of iris visible, size of pupil barely discernible
4* Opaque cornea, iris not discernible through the opacity

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

CONJUNCTIVAL IRRITATION: redness (refers to palpebral and bulbar conjunctivae, cornea and iris)
0 Blood vessels normal
1 Some blood vessels definitely hyperemic (injected)
2* Diffuse, crimson colour, individual vessels not easily discernible
3* Diffuse beefy red

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

* Starred figures indicate positive effect

OBSERVATIONS
- Viability / mortality: daily
- Body weights: day 1 of test (application day) and at termination of test.

NECROPSY
- All rabbits were killed by intravenous injection of T61 into the ear vein.
- No macroscopic organ examination was performed.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7

Results 3

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 5

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.67

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 6

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Results 7

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 9

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritant / corrosive response data:

CORROSION
- A slight diffuse or slight opacity was observed in all animals within 1 hour to 7 days.

Other effects:

TOXICITY
- No acute toxic symptoms were observed in the animals during the test period.

MORTALITY
- No mortality occurred.

COLOURATION
- In the area of applicatlon a slight black to black staining of the cornea, lidhairs and conjunctivae by pigment or colouring of the test item was observed.

BODY WEIGHTS
- The body weight gains of all rabbits was similar.

NECROPSY
- No macroscopic organ examination was performed.

Any other information on results incl. tables

Extrapolation of the results of eye irritation studies in animals to man is valid only to a limited degree. The albino rabbit is more sensitive than man to ocular irritants or corrosives in most cases. Similar results in tests on other animal species may give more weight to extrapolation from animal studies to man.

Applicant's summary and conclusion

Interpretation of results:

other: Category 1 (Serious Eye Damage) according to CLP Regulation (EC) No.1272/2008

Conclusions:

The test item was found to be damaging to the rabbit eye.

Executive summary:

Eye irritation potential of the test item was evaluated in an in vivo experimental study performed according to the OECD Guideline 405 (1987) and EU method B.5 (1984). 0.1 g of undiluted, solid test item was administered to the left conjunctival sac of three female rabbits and was not rinsed. Irritation scores for corneal capacity (maximum: 4), iritis (maximum: 2), conjunctival redness (maximum: 3) and chemosis (maximum: 4) were recorded 1, 24, 48 and 72 hours and 7 and 14 days after application, and irritation scores for each parameter were calculated as mean of irritation scores at 24, 48 and 72 hours. Animals were also monitored for toxic symptoms, mortality, local discolouration and body weight change.

No mortality or symptoms of toxicity were observed during the study period. Body weights of all three animals did not differ significantly. Corneal opacity score was 1 for all animals. Iritis score was 0 for all animals. Conjunctival redness scores were 1, 1.67 and 0 for animals #1, #2 and #3, respectively. Chemosis scores were 0.67, 1.33 and 1 for animals #1, #2 and #3, respectively. All four irritation effects were reversible within the study period (14 days). In the area of application, a slight black staining of the cornea, lidhairs and conjunctivae by pigment or colouring from the test item was observed and persisted until the end of the study period (14 days), and reversibility was not possible to be demonstrated.