Reaction mass of 3,7-bis(diethylamino)phenoxazin-5-ium chloride and 3,7-bis(diethylamino)phenoxazin-5-ium trichlorozincate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

other: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Details in section 13

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Similar substance 01

Molecular formula:

n.a.

IUPAC Name:

Similar substance 01

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

1 males and 2 females
- Body weight at start of acclimatization: 2.7 kg (male) and 2.8-3.0 kg (females)
- Body weight at start of treatment: 2.9 kg (male) and 2.9-3.0 kg (females)
- Age at treatment: 15 weeks
- Acclimatization: 4 days under test conditions after health examination
- Accomodation: individually in stainless steel cages with an automatic cleaning system, equipped with feed hoppers, drinking water bowls and wood for gnawing
- Diet: pelleted standard Kliba 341 rabbit maintenance diet (batch 94/96) ad libitum from Kliba Mühlen AG, CH-4303 Kaiseraugst
- Water: community tap water from Itingen, ad libitum
- Standard laboratory conditions: Air-conditioned with 10-15 air changes per hour and continuously monitored environment with target ranges for temperature of 20 ± 3 °C and for relative humidity between 40-70 % (values above 70 % during cleaning process possible), 12 hours artificial fluorescent light/12 hours dark, music during the light period.

Test system

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

0.1 g of test substance (undiluted)

Duration of treatment / exposure:

single apllication to the conjunctival sac of the left eyelid

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

1 male and 2 females

Details on study design:

TREATMENT:
The eyes of the animals were examined for intactness once, one day prior to test substance administration.Only those animals with no signs of ocular injury or irritation were used on the test.At treatment, 0.1 g of the test article (undiluted) was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The right eye remained untreated and served as the reference control.

SCORING SYSTEM:
CORNEAL IRRITATION
Opacity: degree of density (densest area used for assessment)
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
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 hyperemia, or injection, any of these or combination thereof, iris still reacting to light (sluggish reaction is positive) .................................................................................................................................................................................1
No reaction to light, hemorrhage, gross destruction (any or al 1 of these)............................................................ 2

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

Chemosis:
lids and/or nictitating membranes No swelling ..........................................................................................................0
Any swelling above normal (includes nictitating membranes)...................................................................................1
Obvious swelling with partial eversion of lids ..........................................................................................................2
Swel 1 i ng wi th 1 i ds about hal f cl osed ...............................................................................................................3
Swelling with lids more than half closed ...................................................................................................................4

The maximum scores are:
Cornea : 4 Conjunctival Redness : 3 Iris : 2 Conjunctival Chemosis : 4
Maximum attainable score : 13

When present, corrosion of and/or staining of sclera and cornea by the test
article were recorded and reported

READINGS: 1h, 24h, 48h, 72h, 8d, 15d and 21d after application

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: all animals (24, 25 and 26)

Time point:

24/48/72 h

Score:

0

Reversibility:

other: no ocular effects observed

Results 2

Irritation parameter:

iris score

Basis:

animal: all animals (24, 25 and 26)

Time point:

24/48/72 h

Score:

0

Reversibility:

other: no ocular effects observed

Results 3

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal: 24

Time point:

24/48/72 h

Score:

ca. 2

Reversibility:

fully reversible within: 7 days

Results 4

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal: 25

Time point:

24/48/72 h

Score:

ca. 1.67

Reversibility:

not fully reversible within: 21 days

Results 5

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal: 26

Time point:

24/48/72 h

Score:

ca. 2

Reversibility:

not fully reversible within: 21 days

Results 6

Irritation parameter:

chemosis score

Basis:

animal: 24

Time point:

24/48/72 h

Score:

ca. 2.3

Reversibility:

fully reversible within: 7 days

Results 7

Irritation parameter:

chemosis score

Basis:

animal: 25

Time point:

24/48/72 h

Score:

ca. 1.33

Reversibility:

fully reversible within: 7 days

Results 8

Irritation parameter:

chemosis score

Basis:

animal: 26

Time point:

24/48/72 h

Score:

ca. 2

Reversibility:

not fully reversible within: 21 days

Irritant / corrosive response data:

Reddening and swelling of the conjunctivae was evident in all animals, as was watery discharge. Hyperemia of the scleral blood vessels was noted in two animals. Reddening and swelling of the conjunctivae persisted in one animal until the end of observation, whereas reddening of the conjunctivae was noted ina second animal. Blue or light blue staining of the treated eyes by the test article was observed.No corrosion was observed at any of the measuring intervals.

Other effects:

The body weight of the animals was within the normal range of variability.

Applicant's summary and conclusion

Interpretation of results:

other: Irreversible effects on the eye (Category 1) according to the CLP Regulation

Conclusions:

The test substance produced irreversible effects on the eye.

Executive summary:

The eye irritation test was performed to rabbit eye according to the OECD Guyideline 405.

The substance was found to cause irritation: in particular, the mean values at 24, 48 and 72 hours for not rinsed eyes were 0 for corneal opacity and iris in all animals, equal or higher than 2 for both conjunctival redness and chemosis, in two of three animals tested. In addition, the irritation symptoms were not fully reversible after 21 days of observation. Blue or light blue staining of the treated eyes by the test article was observed.

Therefore, the test substance can be classified in Category 1 due to irreversible effects on the eye, according to the CLP Regulation (EC n. 1272/2008).