Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
19 July 2010 - 22 July 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
see below for details
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
see below for details
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
yes
Remarks:
see below for details
Principles of method if other than guideline:
The temperature and the relative humidity were out of the guideline range between 28 June and 22 July 2010 (max 26.9 °C and 87%). These deviations are considered to have no impact on the outcome of the study and interpretation of the results.
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:
Source: S&K-LAP Kft. 2173 Kartal, Császár út 135, Hungary
Justification of strain: The New Zealand White rabbit is one of the standard strains used for acute irritation toxicity studies.
Animal health: Only animals in acceptable health condition were used for the test. Both eyes of each animal provisionally selected for testing were examined approximately one hour before starting the study. Animals showing eye irritation, ocular defects or pre-existing corneal injury were not used. Number of animals: 3 animals
Age of animals at treatment: ~13-14 weeks old (adult)
Sex: Male
Body weight range at the beginning of the life phase: 3354-3697 g end of the life phase: 3430-3811 g
Date of receipt: 24 June 2010
Acclimatization time: 25 days
Animal identification: The individual identification was by engraved ear tag. The cages were marked with individual identity cards with information about study code, sex, dose group, cage number and individual animal number.
Number of animal room: 632
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 18.2–26.9 °C
Relative humidity: 50–87 %
Housing/Enrichment: Rabbits were individually housed in AAALAC approved metal wire rabbit cages. Cages were of an open wire structure and cages were placed together to allow some social interaction with rabbit(s) in adjoining cages.
Ventilation: 15-20 air exchanges/hour The environmental parameters were recorded twice daily during the study.
Diet: Animals received PURINA Base – Lap gr. diet for rabbits produced by AGRIBRANDS Europe Hungary PLC, H-5300 Karcag, Madarasi road, Hungary, ad libitum.
Water: The animals received municipal tap water, as for human consumption, ad libitum, from an automatic system. The quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives of LAB Research Ltd.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: the contralateral eye of each rabbit served as the control.
Amount / concentration applied:
A single dose of 0.1 mL of the undiluted test item was administered to each animal.
Duration of treatment / exposure:
72 hours
Observation period (in vivo):
The eyes were examined at 1, 24, 48 and 72 hours after treatment. The duration of the observation period was sufficient to identify reversibility or irreversibility of changes. Any clinical signs of toxicity or signs of ill- health during the study were recorded.
At the end of the observation period, each animal was sacrificed by intramuscular injections of CP-Ketamin 10% and CP-Xylazine 2% followed by iv. Euthasol® 40% anaesthesia. Death was verified by checking pupil and cornea reflex, absence of respiration and pulse.
Number of animals or in vitro replicates:
3
Details on study design:
Three male animals in acceptable health condition were selected for the test. Care was taken to select only those animals that had a normal eye condition and any with ocular lesions were rejected.
An initial test was performed using one animal. The test item was instilled into the conjunctival sac of the left eye. The eyelids were held closed for several seconds to prevent the loss of the test item. The contralateral eye served as the control. Immediately after the administration of the test item, an assessment of the initial pain reaction was made. After consideration of the ocular responses produced in the first animal, two additional animals were treated.
The eye irritation scores were evaluated according to the scoring system by Draize (1977) and OECD 405 (24 April 2002).
Individual body weight was recorded at the beginning and end of the experiment.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
0
Reversibility:
other: no effect seen
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
0
Reversibility:
other: no effect seen
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
0
Reversibility:
other: no effect seen
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
0
Reversibility:
other: no effect seen
Irritation parameter:
iris score
Basis:
animal #2
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
0
Reversibility:
other: no effect seen
Irritation parameter:
iris score
Basis:
animal #3
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
0
Reversibility:
other: no effect seen
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #1
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0.33
Max. score:
2
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #2
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #3
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
1
Reversibility:
fully reversible within: 24 hours
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
animal #1
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
3
Reversibility:
fully reversible within: 24 hours
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
animal #2
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
3
Reversibility:
fully reversible within: 24 hours
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
animal #3
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
3
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
1
Reversibility:
fully reversible within: 24 hours
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0.33
Max. score:
2
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
1
Reversibility:
fully reversible within: 24 hours
Irritant / corrosive response data:
The eyes were examined at 1, 24, 48 and 72 hours after the application.
Initial Pain Reaction (IPR) was not observed.
One hour after the application, conjunctival redness (score 2) was observed in two animals and redness (score 1) was noted in one rabbit. Conjunctival chemosis (score 1) was noted in two rabbits, chemosis (score 2) in one animal; conjunctival discharge (score 3) was seen in all rabbits.
At 24 hours after treatment, conjunctival redness (score 1) was observed in one, redness (score 2) in one animal. One rabbit showed conjunctival chemosis.
At 48 hours after treatment conjunctival redness (score 1) was observed in one animal.
At 72 hours after treatment, there were no clinical signs observed.
As there were no clinical signs observed, the study was terminated after the 72 hour observation.
During the study, the control eye of all animals was symptom-free.
Other effects:
There was no mortality observed during the study.
The body weight and body weight change were considered to be normal with no indication of treatment related effect.
There were no clinical signs observed that could be related to treatment.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test substance, applied to rabbit eye mucosa, caused significant conjunctival irritant effects at one hour which were reduced at 24 hours after application. The effects were fully reversible within 72 hours.
According to Regulation (EC) No 1272/2008, the test substance does not require classification as an eye irritant.
Executive summary:

An acute eye irritation study (10/095-005N) of the test item was performed on New Zealand White rabbits in accordance with EU Test Method B.5. 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. A volume of 0.1 mL of the test item was administered as a single dose.

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

Initial Pain Reaction (IPR) was not observed.

One hour after the application, conjunctival redness (score 2) was observed in two animals and redness (score 1) was noted in one rabbit. Conjunctival chemosis (score 1) was noted in two rabbits, chemosis (score 2) in one animal; conjunctival discharge (score 3) was seen in all rabbits.

At 24 hours after treatment, conjunctival redness (score 1) was observed in one, redness (score 2) in one animal. One rabbit showed conjunctival chemosis.

At 48 hours after treatment conjunctival redness (score 1) was observed in one animal.

At 72 hours after treatment, there were no clinical signs observed.

As there were no clinical signs observed, the study was terminated after the 72 hour observation.

During the study, the control eye of all animals was symptom-free.

The general state and behavior 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:

chemosis : 0.00, 0.33, 0.00

discharge : 0.00, 0.00, 0.00

redness : 0.33, 1.00, 0.00

cornea opacity : 0.00, 0.00, 0.00

iris : 0.00, 0.00, 0.00

The test item, applied to the rabbits eye mucosa, caused significant conjunctival irritant effects at one hour which were reduced at 24 hours after application. The effects were fully reversible within 72 hours.

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