Black HM 2482

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 12th to February 15th, 1985

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted May 12, 1981

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

EWG Directive 84/449/ EWG, Amtsblatt der Europaeischen Gemeinschaften L 251, Jahrgang 27, 19. 9. 84. B.5. Akute Toxizitaet Augenreizung/ 109.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

400-920-6

EC Name:

-

Cas Number:

89857-06-7

Molecular formula:

C50 H53 N11 O14 S

IUPAC Name:

5'-[2-(7-{2-[4-(2-{1'-[3-(dimethylazaniumyl)propyl]-6'-hydroxy-4'-methyl-2'-oxo-1',2'-dihydro-1λ⁵-[1,3'-bipyridin]-1-ylium-5'-yl}diazen-1-yl)phenyl]diazen-1-yl}-8-hydroxy-6-sulfonatonaphthalen-2-yl)diazen-1-yl]-6'-hydroxy-3,4'-dimethyl-2'-oxo-1',2'-dihydro-1λ⁵-[1,3'-bipyridin]-1-ylium bis(2-hydroxypropanoate)

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Remarks:

KFM

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoerin AG CH 4474 Fuellinsdorf/ Switzerland
- Age at study initiation: 15 - 16 weeks
- Weight at study initiation: 2.6 - 7.9 kg
- Housing: individually in stainless steel cages equipped with an automatic cleaning and drinking system
- Diet: peIIeted standard KIiba 341, Batch 3/84 rabbit maintenance diet, ad libitum
- Water: comunity tap water from Itingen, ad libitum
- Acclimation period: four days under' Iaboratory conditions after veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 55 ± 10 %
- Air changes: 10 -15 per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescentcIight/12 hours dark, at least 8 hours music/light period

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: A singIe dose was administered to the left eye of each animal. The right eye remained untreated and was used as the reference control. The application volume was 0.1 g per animal.

Duration of treatment / exposure:

72 hours

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 rabbits (1 male and 2 females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
- Time after start of exposure:

SCORING SYSTEM:
CORNEAL IRRITATION
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
- Nacrous area, no details of iris visible, size of pupil barely discernible: 3
- Opaque cornea, iris not discernible through the opacity: 4

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

CONJUNCTIVAL IRRITATION
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 membranes
- No swelling: 0
- Any swelling above normal (includes nictating membrane): 1
- Obvious swelling with partial eversion of lids: 2
- Swelling with lids about half closed: 3
- Swelling with lids more than half closed: 4

TOOL USED TO ASSESS SCORE: eye examination were made with a slit-lamp 30 SL and a Varta Cliptrix diagnostic-lamp

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

Max. score:

4

Results 2

Irritation parameter:

iris score

Basis:

other: all animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

3

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.7

Max. score:

3

Results 5

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Results 6

Irritation parameter:

chemosis score

Basis:

other: all animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

IRRITATION
Test item showed a primary irritation score of 0.6 when applied to the rabbit eye mucosa.
CORROSION
No corrosion of the cornea was observed at each of the measuring intervals.

Any other information on results incl. tables

TOXIC SYMPTOMS/MORTALITY 
No acute toxic symptoms were observed in the animals during the test period, and no mortality occcurred.

COLORATION
In the area of application a severe black discoloration of the cornea and conjunctivae which could be related to effects of the test article was observed. 

BODY WEIGHTS
The body weight gain of all rabbits was similar.

NECROPSY
Due to the results obtained, no macroscopic organ examination was indicated.

Applicant's summary and conclusion

Interpretation of results:

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

Conclusions:

Not irritating

Executive summary:

The substance was tested for eye irritation according to the OECD Guideline 405 (1981). A single dose was administered to the left eye of each animal. The right eye remained untreated and was used as the reference control. The application volume was 0.1 g per animal. The eyes of each animal were examined 1, 24, 48 and 72 hours after administration. Under the conditions of this experiment, test item was found to cause a primary irritation score of 0.6 when applied to the rabbit eye mucosa. In the area of application a severe black discoloration of the cornea and conjunctivae was observed which could be related to effects of the test article. No corrosion was observed at each of the measuring intervals.

The mean value of the scores for each type of lesion, calculated for each animal separately, is the following:

Animal No.    Sex                            Mean 24 -72 hours
                                       Cornea opacity    Iris        Conjunctivae 
                                                                          redness   chemosis 
128             male                   0                   0            1           0
129             female                0                   0            0.7         0
130             female                0                   0            0            0

In conclusion, based on test results the substance does not meet the criteria to be classified as eye irritating, according to the CLP Regulation (EC) No 1272/2008.