Tetrasodium [2-({4-fluoro-6-[(2-{[4-fluoro-6-({5-(hydroxykappaO)-6-[(2-{[2-(hydroxy-kappaO)-5-sulfophenyl] diazenyl-kappaN1}-4,5-dimethoxyphenyl) diazenyl-kappaN2]-7-sulfo-2-naphthyl} amino)-1,3,5-triazin-2-yl] amino} propyl) amino]-1,3,5-triazin-2-yl} amino) benzene-1,4- disulfonato(6-)] cuprate(4-)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experiment start date - 12 May 2006;
Experiment completion date - 07 June 2006;
Study completion date - 07 June 2006.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Tetrasodium [2-({4-fluoro-6-[(2-{[4-fluoro-6-({5-(hydroxykappaO)-6-[(2-{[2-(hydroxy-kappaO)-5-sulfophenyl] diazenyl-kappaN1}-4,5-dimethoxyphenyl) diazenyl-kappaN2]-7-sulfo-2-naphthyl} amino)-1,3,5-triazin-2-yl] amino} propyl) amino]-1,3,5-triazin-2-yl} amino) benzene-1,4- disulfonato(6-)] cuprate(4-)

Cas Number:

882878-51-5

Molecular formula:

C39H28CuF2N14Na4016S4

IUPAC Name:

Tetrasodium [2-({4-fluoro-6-[(2-{[4-fluoro-6-({5-(hydroxykappaO)-6-[(2-{[2-(hydroxy-kappaO)-5-sulfophenyl] diazenyl-kappaN1}-4,5-dimethoxyphenyl) diazenyl-kappaN2]-7-sulfo-2-naphthyl} amino)-1,3,5-triazin-2-yl] amino} propyl) amino]-1,3,5-triazin-2-yl} amino) benzene-1,4- disulfonato(6-)] cuprate(4-)

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Description: Black powder
Stability of test item: Stable under storage conditions.
Storage conditions: At room temperature (range of 20 ±5 °C), in a exsiccator, light protected.

Specific details on test material used for the study:

Identity: FAT 40825/A
Batch number: CHU 297 / BOP 04/05
Purity: Organic part (Na-salt): approx. 83.7 %; All coloured components: approx. 80.54 %; Main component: approx. 59.1 %.
Appearance: Solid, black powder
Storage conditions: At room temperature at about 20 °C
Expiration date: December 31, 2010

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Test system: Young Adult New Zealand White Rabbit, SPF
Number of animals per test: 3 (Animals of both sexes were used)
Age at treatment: 14 - 15 weeks (male) 14 - 15 weeks (females)
Identification: By unique cage number and corresponding ear number.
Acclimatization: Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.

HUSBANDRY: Standard Laboratory Conditions: Air-conditioned with ranges for room temperature 17 - 23 °C, relative humidity 30 - 70 % and approximately 10 - 15 air changes per hour. The animals were provided with an automatically controlled light cycle of 12 hours light and 12 hours dark. Music was played during the daytime light period.
Accommodation: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks and haysticks 4642 (batch no. 77/05) were provided for gnawing.
Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 85/05) provided by Provimi Kliba AG, CH-4303 Kaiseraugst.
Water: Community tap water from Füllinsdorf, ad libitum.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g

Duration of treatment / exposure:

one second and the treated eyes were not rinsed after instillation.

Observation period (in vivo):

Viability/Mortality: Daily from acclimatization of the animals to the termination of test.
Clinical signs: Daily from acclimatization of the animals to the termination of test.
Body weights: At start of acclimatization, on the day of application and at termination of observation. The eyes of each animal were examined approximately 1, 24, 48, 72 hours as well as 7, 10, 14, 17 and 21 days after administration.

Number of animals or in vitro replicates:

3

Details on study design:

The primary eye irritation potential of the test item was investigated according to OECD test guideline no. 405. The test item was applied by instillation of 0.1 g into the left eye of each of three young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours, as well as 7, 10, 14, 17 and 21 days after test item instillation.
The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately.

Grading of Ocular Lesions
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 luster), details of iris clearly visible...................... 1
Easily discernible translucent area, details of iris slightly obscured...................... 2
Nacreous area, no details of iris visible, size of pupil barely discernible...................... 3
Opaque cornea, iris not discernible through the opacity...................... 4

IRIS
Normal...................... 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperemia, 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

CONJUNCTIVAE
Redness (refers to most severe reading of palpebral and bulbar conjunctivae when compared with control eye)
Blood vessels normal...................... 0
Some blood vessels definitely hyperemic (injected)...................... 1
Diffuse, crimson colour, individual vessels not easily discernible...................... 2
Diffuse beefy red...................... 3

Chemosis: lids and/or nictitating membranes
No swelling...................... 0
Any swelling above normal (including nictitating membranes)...................... 1
Obvious swelling with partial eversion of lids...................... 2
Swelling with lids about half-closed...................... 3
Swelling with lids more than half-closed...................... 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0.778

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Very slight corneal opacity affecting the whole area was observed in all animals 1 hour after treatment. No abnormal findings were observed in the iris of any animal at any of the measurement intervals. Moderate reddening of the conjunctivae was noted in all animals at the 1- hour reading and persisted as slight reddening in two animals till 48 hours after application and in one animal until 72 hours after application. No swelling (Chemosis) of the conjunctivae was observed. Slight to moderate reddening of the sclerae was noted in all animals at the 1-hour reading and persisted in two animals till the 48- and in one animal till the 24-hour reading. Slight ocular discharge was noted in all animals at the 1-hour reading. No corrosion of the cornea was observed at any of the reading times.

Other effects:

Slight to marked black staining of the treated eyes produced by the test item was observed in all animals throughout the whole observation period. Black remnants of the test item were observable in the three rabbits from 1 hour to 72 hours after application. No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of the study, the test article is irritant to the eye of rabbits (Stains the eyes ).

Executive summary:

The primary eye irritation potential of the test item was investigated according to OECD test guideline no. 405 in a GLP certified laboratory. The test item was applied by instillation of 0.1 g into the left eye of each of three young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours, as well as 7, 10, 14, 17 and 21 days after test item instillation. The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 1.00, 0.67 and 0.67 for reddening and 0.00 for chemosis for all three animals, respectively. The instillation of test item into the eye resulted in mild and early-onset ocular changes, such as corneal opacity, reddening of the conjunctivae and sclerae and discharge. Slight to marked black staining of the treated eyes was noted in all animals at the 1-hour reading and persisted as slight staining in all animals until the 21-day reading, the end of the observation period for all animals. Black remnants were observed in the eye or conjunctival sac of all the rabbits from 1 hour to 72 hours after application. No abnormal findings were observed in the iris of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No clinical signs were observed. Based on the results of the study, the test article is irritant to the eye of rabbits (Stains the eyes).