This study was designed to be compatible with the procedures indicated by the following 

internationally accepted guidelines and recommendations: 

• OECD Guidelines for the Testing of Chemicals No. 405 "Acute Eye 

Irritation/Corrosion" (adopted 02 October 2012) 

• Method B5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No. 

440/2008 

Initial considerations 

Available information indicated that the test item had the potential to produce severe effects in a 

rabbit eye and to confirm this initial assessment, a Rabbit Enucleated Eye Test was performed 

prior to the in vivo test. The results are given in the Appendix and indicated that the test item 

was unlikely to cause severe ocular irritancy. The negative control data was shared withjCBI jCBI I ~--J 

Experimental Starting Date: 

Experimental Completion Date: 

24 June 2014 

17 July 2014 

Test item characterisation data are the responsibility of the Sponsor. 

All raw data will be retained in Harlan Laboratories Ltd., Shardlow, UK archives. 

METHOD 

A single application of 0.1 mL of the test item was administered to the non-irrigated eye of two 

rabbits. Ocular reactions were recorded 1, 24, 48 and 72 hours after administration. 
The test item produced a maximum group mean score of 10.5 and was classified as a 

mild irritant (Class 4 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and 

Calandra classification system. 

This study was conducted in a facility operating to Good Laboratory Practice within the UK 

national GLP monitoring programme, but the study report has not been audited by the QA Unit. 

No formal claim of GLP compliance is made for this study. This report is an accurate record of 

the study and its outcome. 

A. Poo5 

A Pooles 

Study Director 
I ~1 Report Page3 

TABLES 

Table 1 Individual Scores and Individual Total Scores for Ocular Irritation 

74465 Female 74508 Female Rabbit Number and Sex 

IPR=O IPR=O 

Time After Treatment 1 24 48 72 1 24 48 72 

Hour Hours Hours Hours Hour Hours Hours Hours 

CORNEA 

E = Degree of Opacity 0 0 0 0 0 0 0 0 

F =Area of Cornea Involved 0 0 0 0 0 0 0 0 

Score (E x F) x 5 0 0 0 0 0 0 0 0 

IRIS 

D I 0 0 0 0 0 0 0 

Score (D x 5) 5 0 0 0 0 0 0 0 

CONJUNCTIVAE 

A=Redness 2 1 I 0 2 I I 0 

B = Chemosis I 1 0 0 1 I 0 0 

C =Discharge 1 0 0 0 I 0 0 0 

Score (A+ B +C) x 2 8 4 2 0 8 4 2 0 

Total Score 13 4 2 0 8 4 2 0 

IPR = Initial pain reaction 
Group Total 21 8 4 0 

Group Mean Score 10.5 4.0 2.0 0.0 
that are negligible, or moderate to severe ocular irritants. 

Methods 

Five enucleated eyes, obtained from the New Zealand White strain of rabbit, were maintained at a 

temperature of 32 ±1.5 °C within the superfusion apparatus. 0.1 mL was applied onto the cornea 

of each of three enucleated eyes. The direct effect of the test item on the cornea was assessed by evaluation of corneal thickness, corneal opacity, alteration of corneal epithelium and fluorescein 

uptake, throughout the duration of the test. The data for all endpoints was assessed and an 

estimate of ocular irritancy potential made. 

A further two enucleated eyes remained untreated for control purposes. The negative control data 

was shared with ICBI I 

Results 

Corneal Opacity 

Observation Period (minutes post dosing) 

60 120 180 240 

Cldy Area Cldy Area Cldy Area Cldy Area 

0 0 0 0 0 0 0 0 

Test Eyes 0 0 0 0 0 0 0 0 

0 0 0 0 0 0 0 0 

Control 0 0 0 0 0 0 0 0 

Eyes* 0 0 0 0 0 0 0 0 

Cldy = Corneal cloudiness * = Control data shared with Harlan Laboratories Ltd. Study number 41401640 
Normal Norma

Control Eyes• Normal Normal Normal Normal 

Fluorescein Uptake (240 minutes post dosing) 

Test Eyes Control Eyes* 

Int 0 0 0 0 0 

Area 0 0 0 0 0 

Corneal Swelling (%) (minutes post dosing) 

60 120 240 

Test Eyesa 12.1 8.3 2.2 

Control Eyes*b 10.3 8.8 6.3 

Conclusion 

Following assessment of the data for all endpoints the test item was considered unlikely to have the 

potential to cause severe ocular irritancy in vivo. 

... 

Int = 

a= 

b= 

Control data shared withiCBI 

intensity of fluorescein u!::'lptr:;'lak~e~-------------" 

for each time point the corneal swelling recorded is the mean of three eyes 

for each time point the corneal swelling recorded is the mean of two eyes