Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 25, 2000 - Jun 01, 2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study was conducted according to Good Laboratory Practice (GLP) and followed the OECD guideline for testing of Chemicals 405.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report Date:
3000

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)
Principles of method if other than guideline:
None
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Strain: Albino rabbit, Crl:KBL(NZW)BR, female (f)
Source: Charles River Wiga GmbH, Kifßlegg
Age: about 33 weeks
Weight: 4,31 kg (3,68-5,05)

Test system

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
0,1 g test material
Duration of treatment / exposure:
0.1 g test material was instilled into the conjunctival sac of the left eye of each animal by gently pulling the lower lid away from the eyeball to form a cup into which the test material was placed. The right eye remained untreated and served as control. After instillation, the eyelids were closed for about 30 seconds.
Observation period (in vivo):
1 hour after treatment, after 24, 48, and 72 hours, then daily up to day 8
Number of animals or in vitro replicates:
3 female rabbits
Details on study design:
-- Identification and adaptation
Healthy rabbits were allocated to the study group. The animals were kept in the experimental room for more than 7 days to allow for acclimatization. Each animal was uniquely identified by a tattoo in the ear displaying the animal number. Individual cage cards were affixed to each cage displaying the study number, test material, day of treatment, and animal number.


-- Assignment
3 female rabbits were used for this study.

-- Housing and diet
All rabbits were housed in an air-conditioned room. Lighting was controlled by a timer to provide a 12-hour light and a 12-hour dark regime.
They were kept separately in special rabbit cages (manufacturer: Becker; type K99/30 KU) with plastic grids placed on mobile racks.
Temperature and relative humidity were measured using a thermohygrograph. The room temperature varied from 20 to 22 °C and the relative humidity from 50 to 64 %.

The rabbits received diet for rabbits No. 814, Eberle Nafag ad libitum, and fresh tap water from Makrolon drinking bottles at least three times a week.
The diet is checked periodically according to the specifications of the manufacturer by an independent laboratory, approved by the German government. Analysis includes both qualitative and quantitative evaluation for heavy metals, aflatoxins, pesticides, and antibiotics.
The drinking water is periodically analyzed according to the national regulations for human drinking water.

-- Preparation
Before the application, the test material was ground in a mortar using a pestle.

–- Administration and dose level
To ensure that only rabbits with normal eyes were included in the study, approximately 24 hours before treatment ophthalmological examinations were performed after instillation of a 0.15 % fluorescein solution (Dr. Thilo & Co.) using an ophthalmoscope according to Eisenhut (Basel). Animals with eye defects, injury or irritation were excluded.
0.1 g test material was instilled into the conjunctival sac of the left eye of each animal by gently pulling the lower lid away from the eyeball to form a cup into which the test material was placed. The right eye remained untreated and served as control. After instillation, the eyelids were closed for about 30 seconds.


-- Observation for clinical symptoms
The rabbits were examined for eye irritation and for changes in behavior and general condition 1 hour after treatment, after 24, 48, and 72 hours, then daily up to day 8 of the experimental part.
Eye changes were evaluated according to the DRAIZE- , OECD- and EEC recommendations.

-- Evaluation of eye reactions

- Cornea Scores
A) 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

B) Area of cornea involved not evaluated (Rating according to the DRAIZE method)

- Iris
A) Normal 0
Markedly deepened rugae, congestion, swelling, moderate
circumcorneal 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, hemorrhage, gross destruction
(any or all of these) 2

- Conjunctivae Scores
A) Redness (refers to palpebral and bulbar conjunctivae,
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

B) Chemosis: lids and/or nictating membranes
No swelling 0
Any swelling above normal (includes nictating 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

C) Discharge (Rating according to the DRAIZE method )
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,
and of a considerable area around the eye 3

Mean score of all animals = Mean grading for irritations of cornea, iris, and conjunctivae per time point
Mean score per animal = Mean grading for each animal of irritations of cornea, iris, and conjunctivae (1, 24, 48, and 72 hours after application)
Maximum value = Maximum grading of a sign of irritation within a period

Results and discussion

In vivo

Results
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 1 h, 24 h, 48 h, 72 h
Score:
0
Max. score:
1
Reversibility:
fully reversible
Irritant / corrosive response data:
No signs of irritation were observed at the cornea and iris. The conjunctivae showed redness (score 1) on day 1 of the study. Later on no signs of irritation were observed. The untreated eyes were unchanged.
Other effects:
No signs of clinical toxicity were detected.

Any other information on results incl. tables

Eye alterations

Clinical findings

No signs of irritation were observed at the cornea and iris. The conjunctivae showed redness (score 1) on day 1 of the study. Later on no signs of irritation were observed. The untreated eyes were unchanged.

No signs of clinical toxicity were detected.

Mortality

All animals survived the observation period.

Body weight

Body weight development of the treated rabbits was inconspicuous.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
According to the EC Regulation 1272/2008 the test material CCP-V-1 has not to be regarded as an irritant for the eyes. It has shown a very weak irritating potential.
Executive summary:

Purpose

The purpose of this primary eye irritation assay was to provide information on possible health hazards in case of acute eye contact with a test material and serve as a a rational basis for risk assessment to the eye irritating potential of the test item in man.

Study design

CCP-V-1 was tested for its toxicological properties. For the test of primary eye irritation, 0.1 g of the ground test material was applied into the conjunctival sac of rabbits.

The first examination of the eyes followed 1 hour after instillation. The examinations were performed daily for further 7 days.

Results

No signs of irritation were observed at the cornea and iris. The conjunctivae showed redness (score 1) on day 1 of the study. Later on no signs of irritation were observed. The untreated eyes were unchanged.

Conclusion

According to the EC Regulation 1272/2008 the test material CCP-V-1 has not to be regarded as an irritant for the eyes. It has shown a very weak irritating potential.