Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
March 30, 2007 - Juni 01, 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
, 2004
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
, 2002
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
signed 2005-04-15

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name: BIO 1031/1
Substance type: pure active substance
Physical state: solid
Storage condition of test material: ambient temp., dark, dry, in original container

Test animals / tissue source

Species:
rabbit
Strain:
Himalayan
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Laboratory of Pharmacology and Toxicology GmbH & Co. KG, Germany. The rabbit is a commonly used non-rodent species for acute eye irritation/corrosion tests.
- Age at study initiation: 3.5-4.5 months
- Weight at study initiation:
Animal no. 1: 2.9 kg
Animal no. 2: 3.1 kg
Animal no. 3: 2.9 kg
- Housing: For 8 hours following test item application, the animals were kept singly in restrainers which allowed free movement of the head but prevented a complete body turn, wiping of the eyes with the paws and excluded irritation of the eye by excrements and urine. During the acclimatisation period and after the 8-hour period in restrainers, the animals were kept singly in cages with dimensions of 380 mm x 425 mm x 600 mm.
- Diet: ssniff7 K-H V2333 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany) served as food. The food was available ad libitum.
- Water: ad libitum
- Acclimation period: yes

ENVIRONMENTAL CONDITIONS
- Temperature: 20 +/- 3°C
- Humidity. 30-70%
- Photoperiod: 12 hours light/dark cycle

Test system

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
100 mg of the test item were administered into one eye in each of 3 animals.
Duration of treatment / exposure:
1 hour
Observation period (in vivo):
1, 24, 48 and 72 hours after administration of the test substance
Number of animals or in vitro replicates:
3 rabbits (male)
Details on study design:
The test item was placed into the conjunctival sac of the right 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 in order to prevent loss of the material. The left eye, which remained untreated, served as a control.
As no corrosive or severely irritation properties of the test item were expected, all three animals were treated at the same time.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): 1 hour after administration the eyes were rinsed with portions of 20 mL 0.9% aqueous NaCl solution, each.
The eyes were examined ophthalmoscopically with a slit lamp prior to the administration and 1, 24, 48 and 72 hours after the administration. The eye reactions were observed and registered.
24 hours after administration, fluorescein was applied to the eyes before being examined to aid evaluation of the cornea for possible lesions.

SCORING SYSTEM:
Cornea:
0 = No ulceration or opacity
1 = Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible
2 = Easily discernible translucent area, details of iris slightly obscured
3 = Nacreous area, no details of iris visible, size of pupil barely discernible
4 = Opaque cornea, iris not discernible through the opacity

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

Conjunctivae:
0 = Normal
1 = Some blood vessels hyperaemic (injected)
2 = Diffuse, crimson colour; individual vessels not easily discernible
3 = Diffuse beefy red

Chemosis:
0 = Normal
1 = Some swelling above normal
2 = Obvious swelling, with partial eversion of lids
3 = Swelling, with lids about half-closed
4 = Swelling, with lids more than half-closed

Any further lesions are listed.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal #1
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Max. score:
3
Reversibility:
fully reversible within: 24 hours
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal #2
Time point:
other: overall at 24, 48 and 72 h
Score:
0.33
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal #3
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Max. score:
3
Reversibility:
fully reversible within: 24 hours
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #2
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Max. score:
2
Irritant / corrosive response data:
Under the present test conditions, a single instillation of 100 mg BIO 1031/1 per animal into the conjunctival sac of the right eye of 3 rabbits caused the following changes:
Conjunctival redness was observed in all animals:
- animal no. 1 and 3: 60 minutes (grade 1) after instillation;
- animal no. 2: 60 minutes (grade 2) and 24 hours (grade 1) after instillation.
The corneae and the irises were not affected by instillation of the test item. There were no systemic intolerance reactions.
Reversibility of any observed effect: Changes fully reversible within 2 days

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
SymHelios 1031 is non-irritation to eyes, hence, no labelling is required.