Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 to 25 June 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Compliant to OECD 405 and GLP guideline

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name: Thiazol Blau

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Sándor Ferenc, 2173 Kartal, Császár út 135, HUNGARY
- Age at study initiation: 9 weeks
- Weight at study initiation: 2798-3007 g
- Housing: single
- Diet: PURINA Base – Lap gr. diet for rabbits ad libitum
- Water: tap water ad libitum
- Acclimation period: 12 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.6-22
- Humidity (%): 36-70
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 22 to 25 June 2009

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.1 g
Duration of treatment / exposure:
1 hour
Observation period (in vivo):
72 hours
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): physiological saline solution
- Time after start of exposure: 1 hour


SCORING SYSTEM:
1, 24, 48, and 72 hours, then at 1 week post treatment

1. Conjunctivae

A. Redness (Palpebral and bulbar)
Normal 0
Some blood vessels hyperaemic (injected) 1
Diffuse, crimson colour, individual vessels not easily discernible 2
Diffuse beefy red 3

B. Chemosis
Normal 0
Some 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

C. Discharge
No discharge 0
Any amount different from normal (does not include small amounts
observed in inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs, on considerable area
around the eye 3


2. Iris

D. Values
Normal 0
Markedly deepened rugae, congestion, swelling, moderate
circumcorneal hyperaemia: or injection: iris reactive to light
(a sluggish reaction is considered to be an effect) 1
Haemorrhage, gross destruction, or no reaction to light. 2



3. Cornea

E. 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

F. Area of cornea involved
One quarter (or less), but not zero 1
Greater than one quarter, but less than half 2
Greater than half, but less than three quarters 3
Greater than three quarters, up to whole area 4



Irreversible effects on the eye/serious damage to eyes (Category 1)
If, when applied to the eye of an animal, a substance produces:
— at least in one animal effects on the cornea, iris or conjunctiva that are not expected to reverse or have not fully reversed within an observation period of normally 21 days;
and/or
— at least in 2 of 3 tested animals, a positive response of:
o corneal opacity ≥ 3 and/or
o iritis > 1,5
calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material.


Reversible effects on the eye/irritating to eyes (Category 2)
If, when applied to the eye of an animal, a substance produces:
— at least in 2 of 3 tested animals, a positive response of:
o corneal opacity ≥ 1 and/or
o iritis ≥ 1, and/or
o conjunctival redness ≥ 2 and/or
o conjunctival oedema (chemosis) ≥ 2
o calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material, and which fully reverses within an observation period of 21 days

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
1, 2, 3
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Remarks:
1, 2, 3
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
1, 2
Time point:
other: 24, 48 and 72 hours
Score:
0.33
Max. score:
3
Reversibility:
fully reversible
Remarks on result:
other: redness
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
3
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
3
Remarks on result:
other: Redness
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
1,2,3
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
4
Irritant / corrosive response data:
No abnormalities were detected in the control eyes during the entire study period.
Initial Pain reaction (score 1) was observed in one animal. The rabbit blinked several times in response to instillation of the test item.
One hour after the application, conjunctival redness (score 2) was observed in two animals, conjunctival redness (score 1) in one animal. Discharge (score 1) was found in all animals.
At 24 hours after treatment, conjunctival redness (score 1) was observed in two animals.
At 48 hours after treatment, there were no signs of eye irritancy observed.
At 72 hours after treatment, there were no signs of eye irritancy observed.
Other effects:
no effects on mortality, clinical signs, body weight

Any other information on results incl. tables

INDIVIDUAL SCORES FOR OCULAR IRRITATION

Time

Animal No.

Score of irritation

IPR

Conjunctivae

Opacity of cornea

Iris

Control eye

Other sign

R

CH

D

OD

OE

R

1 hour

1

2

0

1

0

0

0

0

-

0

2

2

0

1

0

0

0

0

-

1

3

1

0

1

0

0

0

0

-

0

24 hour

1

1

0

0

0

0

0

0

-

2

1

0

0

0

0

0

0

-

3

0

0

0

0

0

0

0

-

48 hour

1

0

0

0

0

0

0

0

-

2

0

0

0

0

0

0

0

-

3

0

0

0

0

0

0

0

-

72 hour

1

0

0

0

0

0

0

0

-

2

0

0

0

0

0

0

0

-

3

0

0

0

0

0

0

0

-

R = Redness; OD = Opacity degree of density; CH = Chemosis; OE = Extension of opaque area; D = Discharge; IPR = Initial pain reaction; - = No clinical signs


BODY WEIGHT DATA

Animal Number

Beginning of study
(g)

At the end of study
(g)

Body weight gain
(g)

1

2802

2862

60

2

3007

3208

201

3

2798

2881

83

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test item Thiazol Blau, applied to the rabbit eye mucosa, caused significant conjunctival irritant effects at one hour which were reduced 24 hours after application. The effects were fully reversible within 48 hours.
According to Regulation (EC) No 1272/2008, Thiazol Blau does not require classification as an eye irritant.
Executive summary:

An acute eye irritation study with Thiazol Blau was performed in New Zealand White rabbits. The irritation effects of the test item were evaluated according to the Draize method (OECD No.: 405, 2002).

The test item was placed into the conjunctival sac of the left eye of each animal. The untreated right eye served as control. An amount of 100 mg of the solid test item was administered as a single dose. The eyes of the test animals were rinsed with 0.9% NaCl solution, one hour after application of the test item.

The eyes were examined at 1, 24, 48 and 72 hours after the application.

 

Initial Pain reaction (score 1) was observed in one animal. The rabbit blinked several times in response to instillation of the test item.

One hour after the application, conjunctival redness (score 2) was observed in two animals, conjunctival redness (score 1) in one animal. Discharge (score 1) was found in all animals.

At 24 hours after treatment, conjunctival redness (score 1) was observed in two animals.

At 48 hours after treatment, there were no signs of eye irritation observed.

At 72 hours after treatment, there were no signs of eye irritation observed.

As there were no local or systemic clinical signs observed 72 hours after test item administration, the study was terminated after the 72-hour observation.

 

No abnormalities were detected in the control eyes during the entire study period.

 

The general state and behaviour of animals were normal throughout the study period.

There were no notable body weight changes during the study period.

The animal’s individual mean scores (considering readings at 24, 48 and 72 hours after the treatment) were as follows:

 

Animal Number

Cornea Opacity

Iris

Conjunctivae

Redness

Chemosis

Discharge

1

0.00

0.00

0.33

0.00

0.00

2

0.00

0.00

0.33

0.00

0.00

3

0.00

0.00

0.00

0.00

0.00

 

The test item Thiazol Blau, applied to the rabbit eye mucosa, caused significant conjunctival irritant effects at one hour which were reduced 24 hours after application. The effects were fully reversible within 48 hours.

According to Regulation (EC) No 1272/2008, Thiazol Blau does not require classification as an eye irritant.