6,13-dichloro-3,10-bis-{2-[4-fluoro-6-(2-sulfo-phenylamino)-1,3,5-triazin-2-ylamino]-propylamino}-benzo[5,6][1,4]oxazino[2,3-.b.]phenoxazine-4,11-disulphonic acid, lithium, sodium salt.

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:

other: GLP compliant guideline study, available as unpublished report, no restrictions, fully adequate for assessment.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

6,13-dichloro-3,10-bis-{2-[4-fluoro-6-(2-sulfo-phenylamino)-1,3,5-triazin-2-ylamino]-propylamino}-benzo[5,6][1,4]oxazino[2,3-.b.]phenoxazine-4,11-disulphonic acid, lithium, sodium salt.

EC Number:

418-000-8

EC Name:

6,13-dichloro-3,10-bis-{2-[4-fluoro-6-(2-sulfo-phenylamino)-1,3,5-triazin-2-ylamino]-propylamino}-benzo[5,6][1,4]oxazino[2,3-.b.]phenoxazine-4,11-disulphonic acid, lithium, sodium salt.

Cas Number:

163062-28-0

Molecular formula:

CAS formula: C42 H34 Cl2F2 N14 O14 S4 .xLi. xNa

IUPAC Name:

dilithium(1+) disodium 6,13-dichloro-3,10-bis({[2-({4-fluoro-6-[(2-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)propyl]amino})-5,12-dioxa-7,14-diazapentacene-4,11-disulfonate

Details on test material:

- Name of test material (as cited in study report): FAT 40529/A
- Lot/batch No.: P 1/95
- Storage: Ambient conditions

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Pampalonl s.d.f., Via Pontita 16, 56043 Fauglia, Pisa, Italy
- Age at study initiation: 9-11 weeks
- Weight at study initiation: approximately 2 kg
- Housing: Indlvldually in stainless steel cages measurlng 63 x 48 x 41 cm and equipped with grid floors. Cages were suspended over trays and each tray held an absorbent material which was inspected daily and changed as necessary.
- Diet (e.g. ad libitum): a commercially available anti-biotic free pelleted Iaboratory diet (Altromin MSK, A. Rieper, Bolzano, Italy) ad libitum
- Water (e.g. ad libitum): Animals were offered drlnking water supplied to each cage via water bottles, ad libitum
- Acclimation period: 20 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15-21
- Humidity (%): 45-65
- Air changes (per hr):- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The untreated eye was used as a comparator control.

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

21 days

Observation period (in vivo):

up to 21 days

Number of animals or in vitro replicates:

3

Details on study design:

A 100mg aliquot of the test substance was introduced into the rlght eye of one animal by gently pulling away the lower lid from the eyeball to form a cup into which the test substance was placed. The Iids were then held shut for a few seconds to prevent loss of the test substance. The left eye remained untreated. This animal was dosed as a screen to assess the severity of irritant response. The response to treatment observed was considered not to be unacceptable and did not preclude the treatment of two additional animals, bringing the total to three.
One, twenty four, forty eight, and seventy two hours, seven, fourteen, and twenty one days after dosing the animals were observed under standard conditions and the treated eye examined macroscoptically for damage or irritation to the cornea, iris and conjunctivae' using the untreated eye as a comparator control.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: Average 24, 48 and 72 hours

Score:

1.5

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 2

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: Average 24, 48 and 72 hours

Score:

1.8

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 3

Irritation parameter:

iris score

Basis:

mean

Time point:

other: Average 24, 48 and 72 hours

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 21 days

Results 4

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: Average 24, 48 and 72 hours

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 21 days

Irritant / corrosive response data:

Well defined (scores of 2) to moderate (scores of 3) conjunctival irrltation, iris inflammation (score of 1) and areas of corneal opacity (severity score of 2) were apparent in the treated eye of all three anilmals within one hour of dosing. Both the opaque areas of the cornea and the conjunctivae were discoloured blue with the test substance. The sclera and iris also showed blue discolouration, possibly indicating a degree of local absorptlon of the substance.
Corneal opacity and iris inflammation were maintained durlng the first week following dosing. Conjunctival irritation showed some recovery in two animals within forty eight hours, slight reaction being observed at this time. No conjunctival irritatlon remained in any animal after seven days although responses in the cornea and iris remained at this time and one week later, fourteen days after dosing. BIue staining or discolouratlon of aII regions of the eye also remained.
Iris inflammatlon had disappeared within twenty one days of dosing but areas of corneal opaclty were maintained in the treated eye of all animals. At this time, blue colouration was observed in the areas of the cornea showing opaclty, the sclera and conjunctivae. Iridial staining observed earlier in the study showed some reduction in severity but was still present.

Other effects:

Changes in body weight during the course of the study were not remarkable.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test substance has an irreversible effect on the eye.

Executive summary:

In a GLP compliant eye irritation study, performed according to OECD guideline 405, the test substance (0.1g) was placed in the conjunctival sac of the right eye of three New Zealand White rabbits (RTC 1995). The eyes were examined for 21 days and a mean score was calculated for the observations made at 24, 48, and 72 hours. Scores of 1.5, 1.8, 1, and 2 were observed for conjunctival redness, conjunctival chemosis, iris inflammation, and corneal opacity, respectively. All these effects were fully reversible within 21 days, except for the cornea score.Therefore it was concluded that the test substance had an irreversible effect in the eye.