Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2003-11-20 to 2003-11-28
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study, OECD 405 compliant.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
(version 1992-07-31)
Deviations:
yes
Remarks:
Only insignificant deviations from protocol
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
(Version 1987-02-24)
Deviations:
yes
Remarks:
Only insignificant deviations from protocol
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): Vératrole

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: CEGAV, Saint Mars d'Egrenne, France.
- Age at study initiation: 2 to 4 months
- Weight at study initiation: mean body weight ± standard deviation of 3.5 ± 0.1 kg.
- Housing: individually in polystyrene cages (48.2 cm x 58 cm x 36.5 cm). Each cage was equipped with a food container and a water bottle.
- Diet (e.g. ad libitum): free access to 110 pelleted diet (SAFE, Villemoisson, Epinay-sur-Orge, France).
- Water (e.g. ad libitum): Ad libitum. filtered trhough a FG Millipore membrane (0.22 micron).
- Acclimation period: at least 5 days before the beginning of the study.
- sex: male

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3
- Humidity (%): 30-70%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air.
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2003-11-20 To: 2003-11-28

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: Left eye exposed; right eye served as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml

Duration of treatment / exposure:
one administration
Observation period (in vivo):
0-72 hours
Number of animals or in vitro replicates:
3 males
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): Eyes were not rinsed after application

SCORING SYSTEM:
Conjunctival lesions and discharge
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
- swelling with lids about half-closed ................................................................................................................................3
- swelling with lids more than half-closed ........................................................................................................................4
Redness (refers to palpebral and bulbar conjunctivae, cornea and iris)
- blood vessels normal .........................................................................................................................................................0
- a number of blood vessels definitely hyperemic (injected) .........................................................................................1
- diffuse, crimson colour, individual vessels not easily discernible .............................................................................2
- diffuse, beefy red ................................................................................................................................................................3
Discharge
- absence of discharge .........................................................................................................................................................0
- slight discharge (does not include small amounts normally found in inner canthus) ............................................1
- discharge with moistening of lids and hairs adjacent to lids .......................................................................................2
- discharge with moistening of lids and hairs on wide area around the eye ................................................................3

Iris lesions
- normal ...................................................................................................................................................................................0
- markedly deepened rugae, congestion, swelling, moderate circum-corneal 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, haemorrhage, gross destruction (any or all of these)................................................................2

Corneal lesions
Degree of opacity (area most dense taken for reading)
- no ulceration or opacity ......................................................................................................................................................0
- scattered or diffuse areas of opacity (other than slight dulling or normal lustre), details of iris clearly visible ..1
- easily discernible translucent area, details of iris slightly obscured ...........................................................................2
- nacreous areas, no details of iris visible, size of pupil barely discernible ..................................................................3
- opaque cornea, iris not discernible through the opacity ..............................................................................................4
Area of opacity
- one quarter (or less) but not zero .....................................................................................................................................1
- greater than one quarter but less than a half ..................................................................................................................2
- greater than one half but less than three quarters .........................................................................................................3
- greater than three quarters up to whole area ..................................................................................................................4

Any other lesions observed were noted.


TOOL USED TO ASSESS SCORE:
Conjunctival reactions, iritis and corneal opacification were evaluated daily in each animal.
For the evaluation of corneal opacification (presence or absence, affected area), the eyes were examined under a UV lamp after instillation of one or two drops of 0.5% sodium fluorescein solution (a clear fluorescence is visible in the areas of opacification). This evaluation was performed on day 2 and repeated thereafter whenever necessary.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: overall 24, 48 and 72 hours for each animal
Score:
0 - 0.7
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: Mean individual score at 24, 48, 72h: 0.7/0.0/0.0
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Time point:
other: overall 24, 48 and 72 hours for each animal
Score:
0.3 - 0.7
Max. score:
3
Reversibility:
fully reversible within: within 72 hours
Remarks on result:
other: Mean individual score at 24, 48, 72h = 0.3/0.7/0.7
Irritation parameter:
iris score
Basis:
mean
Time point:
other: overall 24, 48 and 72 hours for each animal
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Remarks on result:
other: Mean individual score at 24, 48, 72h = 0.0/0.0/0.0
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: Overall 24, 48 and 72 hours for each animal
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
other: Mean indivudual scores at 24, 48, 72h: 0.0/0.0/0.0
Irritant / corrosive response data:
A slight chemosis (grade 2), a slight or moderate redness of the conjunctiva (grade 2 or 3) and a whitish purulent discharge were observed in all animals on day 1. Then, a very slight chemosis (grade 1) and/or a very slight redness of the conjunctiva (grade 1) persisted up to day 3 at the latest.
A slight iritis (grade 1) was noted in 1/3 animals on day 1. Mean scores calculated for each animal over 24, 48 and 72 hours were 0.7, 0.0 and 0.0 for chemosis, 0.3, 0.7 and 0.7 for redness of the conjunctiva, 0.0, 0.0 and 0.0 for iris lesions and 0.0, 0.0 and 0.0 for corneal opacity.
Other effects:
no other effects

Any other information on results incl. tables

Animal no. 276. Individual ocular examinations and mean values of the scores recorded for each individual animal (24, 48 and 72 hours).

Animal number

Eye region

Ocular reactions

Scores

Mean irritation score

(1)

Interpretation

(+)

(-)

276

1h

D1

24h

D2

48h

D3

72h

D4

Conjunctivae

Chemosis

2

1

1

0

0.7

(-)

Redness

3

1

0

0

0.3

(-)

Discharge

S

0

0

0

0.0

Iris

0

0

0

0

0.0

(-)

Corneal opacity

Intensity

0

0

0

0

0.0

(-)

Area

0

0

0

0

0.0

Other

*

*

*

*

Fluorescein

/

U

/

/

(1) mean of scores on days, 2, 3 and 4; h = hour; D = day;

(+) = irritant according to EC criteria; (-) = not irritating according to EC criteria;

S = whitish purulent discharge; * = none;

U = fluorescein batch no. H586; / fluorescein not used

Animal no. 281. Individual ocular examinations and mean values of the scores recorded for each individual animal (24, 48 and 72 hours).

Animal number

Eye region

Ocular reactions

Scores

Mean irritation score

(1)

Interpretation

(+)

(-)

281

1h

D1

24h

D2

48h

D3

72h

D4

Conjunctivae

Chemosis

2

0

0

0

0.0

(-)

Redness

2

1

1

0

0.7

(-)

Discharge

S

0

0

0

0.0

Iris

0

0

0

0

0.0

(-)

Corneal opacity

Intensity

0

0

0

0

0.0

(-)

Area

0

0

0

0

0.0

Other

*

*

*

*

Fluorescein

/

U

/

/

(1) mean of scores on days, 2, 3 and 4; h = hour; D = day;

(+) = irritant according to EC criteria; (-) = not irritating according to EC criteria;

S = whitish purulent discharge; * = none;

U = fluorescein batch no. H586; / fluorescein not used

Animal no. 282. Individual ocular examinations and mean values of the scores recorded for each individual animal (24, 48 and 72 hours).

Animal number

Eye region

Ocular reactions

Scores

Mean irritation score

(1)

Interpretation

(+)

(-)

282

1h

D1

24h

D2

48h

D3

72h

D4

Conjunctivae

Chemosis

2

0

0

0

0.0

(-)

Redness

2

1

1

0

0.7

(-)

Discharge

S

0

0

0

0.0

Iris

1

0

0

0

0.0

(-)

Corneal opacity

Intensity

0

0

0

0

0.0

(-)

Area

0

0

0

0

0.0

Other

*

*

*

*

Fluorescein

/

U

/

/

(1) mean of scores on days, 2, 3 and 4; h = hour; D = day;

(+) = irritant according to EC criteria; (-) = not irritating according to EC criteria;

S = whitish purulent discharge; * = none;

U = fluorescein batch no. H586; / fluorescein not used

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the experimental conditions, the Veratrole was slightly irritating when administered by ocular route to rabbits. However, according to the classification criteria laid down in the Regulation (EC) 1272/2008 (CLP) and in Council Directive 67/548/EEC (and subsequent adaptations), the test item should not be classified as irritating to the eyes.
Executive summary:

In a study (2004), the potential of the test item Veratrole to induce ocular irritation was evaluated in New Zealand White rabbits according to OECD (No. 405, 24th February 1987) and EC (92/69/EEC, B.5, 31st July 1992) guidelines.

In the first instance, the test iem was instilled in the eye of a single animal. Since the test item was not severely irritant on this first animal, it was then evaluated in two other animals.

A single dose of 0.1 mL of the undiluted test item was instilled into the left conjunctival sac. The right eye was not treated and served as control. The eyes were not rinsed after administration of the test item. Ocular reactions were observed approximately 1 hour, 24, 48 and 72 hours after the administration.

The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and corneal opacity were calculated for each animal.

A slight chemosis, a slight or moderate redness of the conjunctiva and a whitish purulent discharge were observed in all animals on day 1.

Then, a very slight chemosis and/or a very slight redness of the conjunctiva persisted up to day 3 at the latest.

A slight iritis was noted in 1/3 animals on day 1. Mean individual scores calculated for each animal over 24, 48 and 72 hours were 0.7, 0.0 and 0.0 for chemosis, 0.3, 0.7 and 0.7 for redness of the conjunctiva, 0.0, 0.0 and 0.0 for iris lesions and 0.0, 0.0 and 0.0 for corneal opacity.

Under these experimental conditions, the test item Veratrole is slightly irritant when administered by ocular route to rabbits but the results did not meet the criteria for classification. Therefore Veratrole is not classified for eye irritation according to the Regulation (EC) 1272/2008 (CLP) and the Directive 67/548/EEC.