- 

FINAL REPORT 

STUDY TITLE 

ACUTE EYE IRRITATION STUDY IN 

RABBITS 

STUDY No.: 

DATA REQUIREMENT 

U.S. EPA Health Effects Test Guidelines, OPPTS 870.2400 (1998) 

OECD Gu idel ines for the Testing of Chemicals, Test No. 405 (2002) 

JMAFF 12-Nousan-8147 (2000) 

Official Journal of the European Commun iti es, Methods for the Determination of 

Toxicity and Other Health Effects, Part B.5 (Eye Irritation), 

EYE 

Copy No. 212 

Counci I Regul ation (EC) No. 440/2004 

AUTHOR 

STUDY COMPLETED ON 

16 June 2011 

SPONSOR 

Dow Europe GmbH 

Bachtobelstrasse 3 

CH-88 J 0 Horgen 

Switzerland 

TEST FACILITY 

DEPARTMENT OF SAFETY ASSESSMENT 

ADVINUS THERA PEUTICS LIMITED 

POST BOX No. 58 13, PLOT Nos. 21 & 22 

PEENY A II PHASE, BANGALORE - 560 058 

INDIA 

Page 1150 
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COMPLIANCE WITH GOOD LABORATORY STANDARDS 

Test Substance: (

Study Title: Acute Eye Irritation Study in Rabbits 

Except as noted below, the study described in this report was conducted in compl iance 

with the following Good Laboratory Practice Standards: 

Organisation for Economic Co-operation and Development (OECD) 

ENV/MCICHEM (98) 17 

European Economic Community (EEC) Commission Directive L50/44, Feb.20, 2004 

Japanese Ministry of Agriculture, Forestry, and Fisheries (JMAFF) Nousan Notification 

No. 6283 Agricultural Production Bureau I October, 1999 

Exception to GLP compliance is as listed below: 

• Archiving of raw data at the test fac ility. 

Exception to Test guideline is as listed below: 

• Exception to guideline 405 , for not testing weight of evidence analysis by the 

test facility. 

Study Director 

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16 1."",, ~ 1\ 

Date 

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QUALITY ASSURANCE STATEMENT 

The Study No.: entitled " Acute Eye Irritation Study in 

Rabbits" has been inspected in accordance with the OECD Principles of Good Laboratory 

Practice (as revised in 1997) ENV/MCICHEM (98)17, OECD Paris, 1998, JMAFF GLP 

standards: II-Nousan-No.6283, I October 1999, and EC Directive 200411 O/EC, Official 

Journal of the European Union, L50/44, Feb.20, 2004. 

This study was inspected and findings reported to Management and to the Study Director 

on the dates shown below: 

INSPECTION 

DATE 

28 February 20 I I 

14 March 20 1 I 

25 April 20 I I 

and 

26 April 20 II 

06 June 201 I 

PHASE 

Initiation Phase 

Review of study plan 

In-Life Phase 

Test substance instillation 

(single animal test) 

Reporting Phase 

Draft report review 

Final report review 

REPORTING 

DATE 

28 February 20 II 

21 March 20 1 I 

26 April 201 I 

06 June 20 I I 

Inspections were performed according to the Standard Operating Procedures of the test 

fac ility's Quality Assurance Unit. The final report was inspected against the approved 

study plan and pertinent raw data and accurately reflects the raw data. 

Date: If, .Jv",< , 'VOll 

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Head, GLP Section 

Quality Assurance Unit 

Advinus Therapeutics Limited 

Bangalore 

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1. STUDY PERSONNEL 

The following personnel participated in the conduct of the study. 

Name, Responsibility, 

Section / Department 

Study Director 

Toxicology 

Department of Safety Assessment 

Technical Co-ordinator 

Toxicology 

Department of Safety Assessment 

Cl inical Veterinarian 

Toxicology 

Depaltment of Safety Assessment 

Technical Assistance 

Toxicology 

Department of Safety Assessment 

Document preparation 

Toxicology 

Department of Safety Assessment 

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Function Signatur e with date. 

Overall in-charge 

for the conduct of 

the study and 

report preparation 

Assistance in 

conduct of study 

and report 

preparation 

Veterinary 

services. 

Assistance in the 

conduct of study 

Data entry and 

report compilation 

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TABLE OF CONTENTS 

STUDY TITLE .. ..................................................... ..... .. .... .. ... ... ... ... ... ....... ........ ... ... ...... 1 

COMPLIANCE WITH GOOD LABORATORY STANDARDS ...... .. ......... ............... 2 

QUALITY ASSURANCE STATEMENT .. ... ... ... ... ......... ... ... ... ......... ... ...... ..... .... ... ...... 3 

1. 

2. 

3. 

4. 

5. 

6. 

7. 

8. 

8.1 

8. 1.1 

8.1.2 

8.2 

8.2.1.1 

8.2.2 

8.2 .3 

8.2.4 

8.3 

8.3.1 

8.3.2 

8.3.3 

9. 

STUDY PERSONNEL ... .. .............. ........................................................ ...... 4 

SUMMARy ............................ ..... ...... .......... ..................... ............ ................ 7 

STUDY DETAILS ..................... ... ............................ .......... ..... ............. ... .. ... 8 

OBJECTIVE ................................................................ .. ......................... ... ... 9 

JUSTIFICATION FOR SELECTION OF TEST SYSTEM ... .. ...... .......... ... 9 

JUSTIFICATION FOR ROUTE OF ADMlNISTRATION ... .... ............ ...... 9 

TEST PROCEDURE GUIDELINES .. .. ................................. ... ................... 9 

MATERIALS AND METHODS ... ... .. ....... .. .......... ... ... ... .... .... ...................... 9 

Materia ls ........................ .... .................................... ....................................... 9 

Test substance Information .............. ..... ...... .. ......... ........................... ............ 9 

Test System ............... .............. ...... ... ... .... .. .... ......... ...... ... .. ....... .. ................. II 

Methods .. .... .. ....... ... ........... ... ... ...... ... ... ................................................ ........ II 

Husbandry ............................................. ........ .... .. ............ ... .. .................. ..... II 

Test Substance Instillation ... ....... ..... ... ..................... ......................... .......... 12 

Animal Restraint ... ... ....... .. ... ...... .............. ...••............................•................. 13 

Stepwise exposure of Animals ............ ....... ......... .. ... ........... .. .... .. ..... ........... 13 

Observations ............. ... ... ... ..... ...... ........................ ...................................... 13 

Toxic Signs and Pre-Terminal Deaths ....... ... ....................... ... .... ..... ...... ..... 13 

Body Weights .... .. ... .. .... .. ... ... .......... .... .............. ................. .......... ...... .......... 13 

Eye Scoring .................. ... ....... ....... ...................... ..... .... .. ....... ............. ... ...... 13 

RESULTS ............ .................. ....................... .. .. .. .. .... .. .... .. ... ..... .... .... ...... .... 14 

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9. 1 Toxic signs and pre-terminal deaths ........................................................... 14 

9.2 Body weights ............................................................. ................................. 14 

9.3 Eye reactio ns ...... ......................................................................................... 14 

10. CONCLUSION ........................... ................................................................ 14 

II. ARCH IVING .............................................................................................. 14 

12. REPORT DISTRIBUTION ........................................................................ 15 

13. TABLES ..................................................................................................... 16 

TABLE I. Body Weight, Toxic Signs and Pre-terminal deaths ............. 17 

TABLE 2. Individual Eye Irritation Scores ............................................ 18 

TABLE 3. Mean Eye Irritation Scores .................................................... 19 

14. APPENDICES ............................................................................................ 20 

APPENDIX I. Certificate of Analysis .................................................... 2 1 

APPENDIX 2. N umerical Scoring System (Grades of Ocular lesions) .22 

APPENDIX 3. Study Plan ...................................................................... 24 

APPENDIX 4. Deviation fro m the Approved Study Plan ..................... .43 

15. ANNEXURES ............................................................................... ............. 44 

ANNEXURE I. Analysis Report - Anima l Diet Sample ........................ .45 

ANNEXURE 2. Analysis Report - Water Sample ................................... 46 

ANNEXURE 3. GLP Certificate - Germany .................................... ...... .47 

ANNEXURE 4. GLP Certificate - The Netherlands ............................... .48 

ANNEXURE 5. GLP Certifi cate - India .................................................. 50 

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2. SUMMARY 

The acute eye irritation/corrosion study in rabbits was conducted to eva luate 

the eye irritation potenti al of Under the conditions of 

this study, caused conjunctival irritation and cornea l 

opacity in the rabbit eye, whi ch clea red by 72 hours. 

One hundred mg (0.1 g) of the test substance was insti lled into the conjunctival 

sac of the left eye of the animal after gently pulling the lower lid away from the 

eyeba ll. The lids were gently held together for about one second, in order to 

minimize loss of the test substance. The right eye of each rabbit remained 

untreated and served as the reference control. The ocular lesions were 

evaluated at I, 24, 48 and 72 hours according to the scale fo r scoring ocular 

lesions. There were no toxic signs or animal death . All rabbits appeared healthy 

and gained body weight through the observation period. 

Under the conditions of this study, the mean eye irritation scores were as 

follows: 

Observations Mean score 

at post Conj unctiva Iris Cornea Tota l mean 

instillation score 

I hour 6.00 0 0 6.00 

24 hours 6.00 0 6.67 12.67 

48 hours 2.67 0 3.33 6.00 

72 hours 0 0 0 0 

The individua ll y determined mean irritation scores for each an imal for cornea l 

opacity, iris lesion, conj unctiva l redness and conjunctiva l chemosis for the 

24, 48 and 72-hour intervals are : 

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Animal No. 

(Sex) 

RB8767 (M) 

RB8768 (M) 

RB8769 (M) 

Corneal 

Opacity 

0.7 

0 

0 

Iris Lesion Conj unctival Conjunctival 

Redness Chemosis 

0 1.3 0.7 

0 0.3 0 

0 0.3 0 

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3. STUDY DETAILS 

Study Title 

Test substance 

Study Number 

Sponsor 

Monitoring Scientist 

Test Facility 

Study Schedule 

Study initiation date 

Experimental starting 

date 

Accl imatization 

Treatment 

Observations 

Experimental 

completion date 

Draft report 

Study completion date 

·EYE 

Copy No. 212 

Acute Eye Irritation 

Study in Rabbits. 

(

Dow Europe GmbH 

Bachtobelstrasse 3 

CH-8810 Horgen 

Switzerland 

Donna Anderson 

The Dow Chemical Company 

Toxicology & Environmental Research & 

Consulting (TERC) 

Building 5 

727 Norristown Road, P.O. Box 904 

Spring House, PA 19477-0904 

Email: DonnaAnderson@dow.com 

Department of Safety Assessment 

Advinus Therapeutics Limited 

Post Box No. 58 I 3, Plot Nos. 21 & 22 

Peenya II Phase, Bangalore 560 058, INDIA 

07 March 201 I 

09 March 2011 

Start: 09 March 201 I 

End: 13 March 2011 (single animal) and 

14 March 2011 (additional two animals) 

Start: 14 March 20 II (single animal) 

15 March 2011 (additional animals) 

Start: 14 March 2011 End: 18 March 2011 

18 March 2011 

25 May 2011 

16June2011 

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4. OBJECTIVE 

The objective of this acute eye irritation study in rabbits was to provide 

information on the potential for irritation arising from a single instillation of a 

test substance into the eye. 

5. JUSTIFICA TION FOR SELECTION OF TEST SYSTEM 

The rabbit is the system of choice because, historically, it has been the 

preferred and most commonly used species for primary eye irritation tests. 

6. JUSTIFICATION FOR ROUTE OF ADMINISTRATION 

The ocul ar route of administration was used because it is recommended in the 

referenced gu ide lines and because human exposure may occur via this route. 

7. TEST PROCEDURE GUIDELINES 

» U.S. EPA Health Effects Test Guidelines, OPPTS 870.2400, Acute Eye 

Irritation, August 1998. 

» OECD Guideline for Testing of Chemicals, Section 4, Number 405. 

"Acute Eye Irritation / Corrosion" adopted 24th April 2002. 

» JMAFF 12-Nousan-8147, November 2000. 

» Official Journal of the European 

Determination of Toxicity, Part 

2004/73/EC, 29 Apri l 2004. 

Communities, Methods for the 

8.5 (Eye Irritation), Directive 

8. MA TERIALS AND METHODS 

8.1 Materials 

8.1.1 Test substance Information 

(As furnished by the Sponsor) 

Test substance / 

Item 

Lot No. 

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Manufactured by 

(Name and 

Address) 

Suppli ed by 

(Name and 

Address) 

Date of 

manufacture 

Date of expiry 

Purity as per 

Certificate of 

Analysis 

Physical 

appearance 

Storage conditions 

The Dow Chemical Company 

Midland, MI 48674 USA 

The Dow Chemi cal Company 

Midland, MI 48674 USA 

March 20 10 

06 March 20 12 

Off-white solid 

: Ambient (+ 18 to + 36°C) 

The test substance, identified as was received 

on 03 February 20 I I and was further identifi ed as 008-26 by test 

fac ility, Reference Number dated 07 February 20 II. The test substance 

is a solid and is stored at ambient cond ition (+ 18 to +36°C). 

Documentation of the methods of synthesis, fabrication, or derivation 

of the test substance is retained by the Sponsor. 

Determination of the stability, identity, strength, purity and 

composition of the test substance is the responsibility of the study 

Sponsor. The fo llowing information related to the characterization of 

the test substance was provided by the sponsor. 

Information as provided by Sponsor are as follows: 

pH : 6.47 (at 1% w/v in aq ueous solution, ana lysed in-house). 

Specific gravity / density: Not App licable. Test substance is a solid. 

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8.1.2 Test System 

Animals 

Source 

No. of rabbits 

Age at treatment 

Date of birth 

Body weight range at 

treatment 

Identification 

Rabbit accession 

numbers 

Accl imati zation 

8.2 Methods 

8.2.1.1 Husbandry 

Conditions 

New Zea land White Rabbits 

Inhouse Bred (Outbred) 

Toxicology, Department of Safety Assessment 

Advinus Therapeutics Limited 

Bangalore 560 058, INDIA 

3 male rabbits 

7 - 8 months 

26 July 20 10 to 30 July 20 10 

2.61 to 2.94 kg 

By animal accession number, cage label, 

crystal violet colour body marking. 

During acclimatization the animals were 

identifi ed by ear tattooing numbers. 

RB8767 to RB8769 

One rabbit (RB8767) was acclimatized for five 

days whil e the other two (RB8768 and 

RB8769) rabbits were acc limatized for six days 

under laboratory conditions after physical 

examination. 

Animals were housed under standard laboratory conditions, air cond itioned 

with adeq uate fresh a ir suppl y (12-15 air changes/hour). Environment: 

temperatu re 2 1-23°C, relative humidity 59-67%, with a 12 hour li ght and 12 

hour dark cycle. The maximum and minimum temperature and relative 

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humidity in the experimental room was recorded once dail y. The relative 

humidity in the experimental room was calculated from dry and wet bulb 

temperature recordings. 

HOllsing 

The rabbits were housed individua lly in rabbit cages (approx. size: L 65 x B 65 

x H 45 cm) with noryl shallow cage body and faci lities for pelleted food 

(stainless steel feed hopper) and drinking water (750 mL markrolon bottle fitted 

with sipper tube). The litter coll ection tray (noryl waste tray) was changed dail y 

(except on Sunday). The feed hoppers were changed once a week. 

Diet ad libitum 

The animals were offered rabbit feed manufactured by Pranav agro Industries 

Ltd, Sangli Maharashtra, India. The diet analysis report is enclosed as 

Annexure I. 

Specific contaminants (heavy metals, organochlorines and organophosphates) 

in the diet are analysed periodicall y and the report are included in the study file. 

Water ad libitum 

Deep bore-well water passed through activated charcoal fi lter and exposed to 

UV rays in Aquaguard on-line water filter-cum-purifier, manufactured by 

Eureka Forbes Ltd., Mumbai-40000 I, India was provided to an imals in 

Markrolon bottles with stainless steel sipper tubes. The water bottles were 

replenished once dail y and the water bottles were changed once a week. The 

water anal ysis repOit is enclosed as Annexure 2. 

8.2.2 Test Substance Instillation 

Prior to insti ll ation, both eyes of each rabbit were examined with the aid of a 

pen li ght for irritation and ocu lar defects. The eyes were then checked for 

opacity using ophthalmic fluorescein sod ium strips. The eyes were then 

irrigated w ith enough physiological sali ne (0.9% NaCI) to remove any excess 

fluorescein (approximately 30 seconds). Using an ultraviolet lamp, the eyes 

were checked and eva luated for pre-existing corneal damage. No cornea l 

defects were observed in any of the animals. 

One hundred milligrams (O.l g) of the finel y ground test substance was placed 

in the everted lower lid (conj unctival sac) of the left eye of each rabbit. 

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The eye lids were gently held together for about one second, in ord er to 

minimize loss of the test substance. The right eye of each rabbit remained 

untreated and served as the reference control. 

8.2.3 Animal Restraint 

Each rabbit was restrained using an Elizabethan collar fixed around the neck. 

One rabbit RB8767 was restrained for 72 hours and the other two rabbits 

(RB8768 and RB8769) were restrained for 48 hours post-instillation. 

8.2.4 Stepwise exposure of Animals 

A si ngle rabbit was initially exposed to the test substance. Since the test 

substance was not corrosive, the test was completed using two add itional 

an imals. 

8.3 Observations 

8.3.1 Toxic Signs and Pre-Terminal Deaths 

The rabbits were observed for toxic signs and pre-terminal deaths four times (at 

hourly interva ls) on the day of test substance instillation and once dail y till 72 

hours post instill ation. 

8.3.2 Body Weights 

Body we ights were recorded at the start of acclimatization, Day I of treatment 

Gust before test substance instill ation) and at study termination. 

8.3.3 Eye Scoring 

Ocular lesions were evaluated with the illumination of a white light source at 

intervals of approximately I, 24, 48 and 72 hours post-instillation of the test 

substance according to the "Scale for Scoring Ocular Lesions" (Draize, 1977). 

t he fluorescein dye evaluation procedure described in Section 8.2.2 was used 

to examine the treated eye of one rabbit (# RB8767, which showed corneal 

opacity at 24 hours) at the 24- and 48-hour observation. At the 72-hour 

observation, this rabbit had no opacity and all its eye scores were O. Hence, 

fluorescein was not used and the experiment was terminated. The other two 

rabbits had no corneal opacity, so their eyes were not observed using 

ophthalmic fluorescein. 

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Individual scores were recorded for each rabbit. All scores were calculated as 

described in the "Scale for Scoring Ocular Lesions", and the total possible 

points (out of 110) were tallied. [n addition to the observation of the cornea, iris 

and conjunctivae, any other lesions that were observed were noted. To aid in 

the interpretation of data, the average score for all rabbits at each scoring period 

was calculated (Appendix 2). 

9. RESULTS 

9.1 Toxic signs and pre-terminal deaths 

Refer to Table I 

There were no toxic signs or pre-terminal deaths. 

9.2 Body weights 

Refer to Table I 

The body weights of all rabbits increased slightly through the observation 

period. 

9.3 Eye reactions 

Refer to Table 2 and 3 

The summary of corneal, iri S and conjunctival reactions (Individual Eye 

irritation scores) and total scores are presented in Table 2 and mean eye 

irritation scores are presented in Table 3. 

10. CONCLUSION 

Under the conditions of this study, caused conjunctival 

irritation and corneal opacity in the rabbit eye, wh ich cleared by 72 hours. 

11. ARCHIVING 

The original signed final report wi II be sent to the Sponsor and one signed final 

report will be retained by Advinus. The original raw data, documentation, 

records, the original study plan, and final report generated as a result of this 

study will be sent to the Sponsor for retention following signing of the final 

report. Copies of the same (authorized by the Study Director) will be retained 

by Advinus. Advinus wi II maintain these records for a period of at least fi ve 

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years. After this time, the Sponsor of the study will be offered the opportunity 

to take possession of the records or will be charged an archiving fee for 

continued archiving by Advinus or to dispose of the raw data in consultation 

with the Sponsor. A representative sample of approximately one gram of the 

test substance has been sent from the test substance stores to the archives in the 

test facility before the first dispensing of test substance to studies. This sample 

shall be stored for a period of 2 years from the date of this final report or till 

next GLP inspection, whichever is later, however not beyond 5 years. 

In the event of any inspection by regulatory authorities, seeking original raw 

data for verification, the sponsor would be responsible either to send the 

original raw data to Advinus Therapeutics Limited or to the regulatory agencies 

for verification. 

12. REPORT DISTRIBUTION 

The final report (original copies) will be distributed as follows: 

Sponsor: One signed final report in original (Copy No. 2/2) and one soft copy 

of the report in PDF format. 

Archives: One signed final report in original (Copy No. 1/2). 

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13. TABLES 

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TABLE 1. Body Weight, Toxic Signs and Pre-terminal deaths 

Rabbit 

No. 

RB8767 

RB8768 

RB8769 

M: Male 

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Copy No. 212 

Sex 

M 

M 

M 

Body weight (kg) 

At Start of At At 

acclimatization treatment Termination 

2.57 2.61 2.69 

2.77 2.88 2.92 

2.82 2.94 2.99 

Toxic PreSigns terminal 

deaths 

None None 

None None 

None None 

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TABLE 2.Individual Eye Irritation Scores 

Rabbit Elapsed CONJUNCTIVA IRIS COR

No. Time Redness Chemosis Discharge Tissue score Pupil Tissue Opac ity Area o

(hours) (0-3) (0-4) (0-3) (0-20) (0-2) score (0-4) opacity 

A B C (A+B+C) x 2 D (0- 10) E (1-4) 

Dx 5 F 

RB8767 I I 0 2 6 0 0 0 NA 

24 2 I 2 10 0 0 I a 4 

48 2 I I 8 0 0 I a 2 

72 0 0 0 0 0 0 0 NA 

RB8768 I I 0 2 6 0 0 0 NA 

24 I 0 I 4 0 0 0 NA 

48 0 0 0 0 0 0 0 NA 

72 0 0 0 0 0 0 0 NA 

RB8769 I I 0 2 6 0 0 0 NA 

24 I 0 I 4 0 0 0 NA 

48 0 0 0 0 0 0 0 NA 

72 0 0 0 0 0 0 0 NA 

NA: Not applicable; a = ophthalmic fluoresce in sodium strips used to aid in evaluating the extent of corneal damage 

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TABLE 3. Mean Eye Irritation Scores 

Observations 

at post 

instillation 

1 hour 

24 hours 

48 hours 

72 hours 

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Conjunctiva Iris 

6.00 0 

6.00 0 

2.67 0 

0 0 

Mean score 

Cornea Total mean 

score 

0 6.00 

6.67 12.67 

3.33 6.00 

0 0 

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14. APPENDICES 

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APPENDIX 1. Certificate of Analysis 

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P~3ofl!l 

20fiS 

CERTIfICATE OF ANALYSIS FOR TESTIREFERENCElCQNTROL SUBSTANCES 

TITLEIOBJECn\'E: QlMlICICtiutiOn or for U6(; IU' TCIl! and Reference 

Subs/mot 

TESTIRE FERENC£lCQ,"'TROL S UBST ANCt;: 

DESCRIPTION: 

SfRUCTlJRE: 

MOLECULAR WEIGIIT

SnroV TNITfATIONOAT£: ISJune20 10 

METIIDDS USEO: 

PURJlY: Gu d1TO~IOgnpby willi flame ioniution dt:Ieaioo (GC·FID) and Kul Fi$(:beT tilr.l.liQCl, 

IDENTIFICATION: Fourio:r TTJnsform-lntnred (FT·IR) specll'O$(:Opy &lid gas tlu-omaloi;f8phy-mau specuomcvy 

(GC-MS) 

RESULTS AND CONCLUSIONS; 

SIf'u(:(uraJ contirmaLion of l>lc l<'IIt ~Irnal was pcrfonncd by FT·1R and GC·MS. 

PURITY DETERMJNAll0N 

Rl'Sult; alltreorrccting (or !he Kul , W1lcr result<. 

onmR (Specify): 6$$ ppm water by Kar] Fischer tirration. No impurities were ckfttted at;:: 0.1% by 

GC-fTD anal)'$il. 

CALCULA nONS: 

Arc_ Percent; '" (GC·FTD) Interm! Standw; NlA fu:temal Sbnthtd: NlA 

Other (uplain): Karl Fischer titr.u ion 

All origin.! IlIW dna associated with thilltudy were archived in the Sporuor racility a«:hivt . 

Only descriptive ltati$tics wert: Died unle.n otherwise noted in the mullS. Thif flooy was condl.lcletl i.n lCCordmcc with the 

Good Labor.uory Practice StandMd. 40 ern. Pan 792 unteu otherwi~ noted. Due (0 LIlc short \(Tm natule orlbe study, th~ 

s(Rbility orlbe t<:JI Sl.IbsLDnc:c 1Uldcl llorggc conditions per 40 CFN. Pcut 792.IO$(c) (U l~e test lite wu nol verified. 

Comple-tion Dalt: 

7(.,., /;w r(l 

'Dati: 

__ 11. <kL ""~ __ _ o.<,rSTUDY DIR£CTOR AND TESTINC FACILITY ADDRESS: 

Impact Analytical, 1910 W. St Andrews Rd. Midland, M14R640 

THIS IS A TRUE COpy OF 

THE ORIGIN/\, __ D.; ~A 

CAe. 12- 2.3- ID 

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APPENDIX 2. Numerical Scoring System (Grades of Ocular lesions) 

(DRAIZE,1977) 

1. CONJUNCTIVAE 

SCORES 

(A) Red ness (refer to palpebral and bulbar conjunctivae 

excl uding corn ea and iris) 

Vessels normal ..................................................................... 0 

Vessels definitely inj ected above normal ............................ .. 

More diffuse, deeper crimson red, individual vessels 

not easily discernible .... .............................. .... .... ................... 2 

Diffuse beefy red .................................. ...... ........................... 3 

(B) Chemosis 

Normal ... ............ .. ... .. ...... ... ... ..... .... ... .... .. ..... .. ........................ 0 

Any swelling above normal (including nictitating 

membrane) ............................................................ .. ... ............ I 

Obvious swelling with partial eversion of lids .................... .. 2 

Swe lling with lids about half closed ...................................... 3 

Swelling with li ds half c losed to complete ly closed ............. 4 

(C) Discharge 

No discharge .................................................................... ...... 0 

Any amount different from normal (does not inc lude 

small amounts observed in inner canthus of norma l 

an imals) .. ..... .... ...... .. ... ..... .. .... ... .. ........ .. ... ............................. . 

Discharge with moistening of the lids and hairsjust 

adjacent to lids ................................................................ ....... 2 

Discharge with moistening of the lids and hairs a 

considerab le area around the eye ...... .. ................................... 3 

TOTAL SCORE = (A + B + C) x 2 MAXIMUM TOTAL=20 

2. IRIS 

(D) 

Normal ........ .. ... ....... ...... ........ ............... .................................... . 

Folds above normal, congestion, swelling, 

Circumcornea l injection (anyone or all of these or 

Combination of any thereof) iris still reacting to light 

(sluggish reaction is positive) .......... .. ...................................... . 

No reaction to light, haemorrhage, gross destruction 

(anyone or all of these) .......... .... .. .............. .. .......... ................ .. 

SCORES 

o 

2 

TOTALSCORE = Dx5 MAXIMUM TOTAL = 10 

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APPENDIX 2 eontd. Numerical Scoring System (Grades of Ocular lesions) 

3. CORNEA 

(E) 

Degree of Opacity (most dense area used) 

No opacity ................................................. ............................ . 

Scattered or diffuse areas- details of iris clearly 

visible .................................................................................... . 

Easily discernible translucent areas, details of iris 

Slightly obscured .......................... ..... ........ ......................... ... . 

Opalescent areas, no details of iris visible, size of 

pupil barely discernible ................................... ...................... . 

Opaque, iris not discernable through the opacity ................... . 

(F) 

Area of Cornea Involved 

One quarter (or less) but not zero ......................................... .. 

Greater than one quarter but less than half ............................ . 

Greater than half but less than three quarters ....................... .. 

Greater than three quarters, up to whole area ........................ . 

SCORES 

o 

2 

3 

4 

SCORES 

I 

2 

3 

4 

THE TOTAL SCORE = (E x F) x 5 MAXIMUM TOTAL = 80 

EYE 

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MAXIMUM TOTAL SCORE POSSIBLE = 110 

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APPENDIX 3. Study Plan 

STCDYPLAK 

STlJ""DY PLAl, No. : SPTEST ITEM CODE 

BY TEST FACILITY : 0 08-26 

STUDY CODE E"\'E 

ACulE EYE IRRlTATIO~ STL"DY 

I~RABBITS 

S P- L'\c 

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TEST ITE::>I:

STUDY Ko.: 

SPOl'SOR 

D ow Europe GmbH 

Bachtobelstrasc;e 3 

CH-8810 Horgen 

Switzet-land 

TEST FACILITY 

DEPARTMENT OF SAFETY ASSESSMENT 

ADVINUS THERAPEUTICS LIMITED 

POST BOX N o. 5813. PLOT Nos. 2 1 & 22 

PEENY A II PHASE. BANGALORE - 560058 

D-UIA 

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CO;\TE;\TS 

1. STUDY DETAILS ......................................... . .................. 4 

1. QUALITY ASSURANCE ............................................................ 5 

3. GLP COMPLiAl'\CE ................................................................... 5 

4. TEST PROCEDURE GUIDEUNES ............................................ 6 

5. AMENDMTh"T PROCEDURES ........ . . ..... .. .............. ... ........ 6 

6. SAFETY PRECAUTIONS ............................. . . ..................... 6 

7. OBJECTIVE ................................................................................ 7 

8. JUSTITiCA TION OF SELECTION OF TEST SySTEM ............... 7 

9. JUSTmCA TION FOR ROUTE OF ADMINISTRi\' TION ............ 7 

10. MATERIALS Al'\;n METHODS ................................................. 7 

10. 1 Materials ........................................................................................ 7 

10. 1.1 

10.1.2 

10.2 

10.2. 1 

10.2.2 

10.2.3 

10.2.4 

10.3 

10.3. 1 

10.3.2 

10.3.3 

11. 

1\.1 

12. 

13. 

Tes1 '!.)"')tC'Ill .............................................................................. 7 

Test !)lJb5tance. information .. ' ........... , .............................................. 8 

Methods .......................... ... ............................................................ 9 

.................. ............................................................ 9 

Test !)ub~ta ce. instillat ion .. , ........ " ......... , ..................... " .............. , 10 

A.n.in1ai res1raint ................................ ............................................ 10 

Stepwise expos\U'e of 311illl.c.11s ....................................................... 10 

Observations ................................................... , ........................... , 11 

('li.lliea! o~ervati llS .. ., ........... , ......... ., ....... ... ., ............................. 11 

Body weights .................. ......... ... .............................. ................ 11 

Eye scoring ... .............. .. ............................. ............................ 11 

RESULTS .................................................................................... 13 

Data cOIltpilatioIl ......... ............................. , .................................. 13 

REPORTIl\G ........ ....................................................................... 14 

ARCHIVING ................... .......................................................... 15 

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14. STUDY PLAl'l DISTRIBUTION ................................................. 15 

15. REPORT DISTRlBUTION ....................................................... 16 

16. REFERENCES ......................................................................... 16 

17. AGREE1viENT ............................................................................ 17 

APPENDIX I. PROCEDURES FOR USE OF ANESTHETICS DURING 

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OCULAR IRRITATION TEST IN RABBITS ...................... 18 

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L STl:DY DETAILS 

Study Title 

Te~t substnnce 

Study Kumber 

Test Facility 

Sponsor 

Monitoring Scientist 

Study Pcnolluel 

Study Director 

Tecluuca! 

Co-ordin ... 'ltor 

Clinical Veterinarian 

Study Schedule 

Study initiation date 

Experi.mental staIting 

date 

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Acute Eye Il1"itatioll Study 

in ~,bbit . 

(

07694 

Department of Safety Ass¢s,smeut 

Advi..llus Therapeutics Limited 

Post Box No. 5813, Plot Nos. 2 1 & 22 

Peonya II Phase, Bangalore 560 058 

IN"DIA 

Dow Etllupe GmbH 

Bachtobelstrasse 3 

CH-88 10 Borgen 

Switzerland 

DOIUla Anderson 

TIle Do\v Chem..ical Company 

Toxicology & Envi.roJUuental Resetlrch & 

COlm.lting (TERC) 

Building 5 

727 Nonlstowll Road. P.O. Box. 904 

Spring House. PA 19477-0904 

Em.ail : DOIUlaA.1lderson(@dow.COlll 

P. J. Prakash, M.Sc. 

S. B. Mohan Kutu<"1l", M .Sc. 

S. Rmnesh. B.v.Sc. & A.H. 

07 March 20 II 

09 March 20 11 

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Ace 1ll1l<1tizat ion 

TrcatJllcnt 

Obse"rvatiollc,: 

Experimental 

completion date 

Draft RepOlt 

Study completion d'\te 

Stmt : 09 March 20 II 

End : 13 March 201 1 (single anuml) and 

14 March 2011 (additional two anulk,lls)' 

Start: 14 March 2011 (single anink,l) 

IS March 2011 (addition,,1 allumlis), 

Start: 14 March 2011 

L.,te,t by IS April 2011 

Latest by 30 April 2011 

L.,t"t by 31 May 2011 

*: \Vi1I be included if the test lIbstallce proves to be llo11-con-o sive and I or 

not a severe in"itant depending on the single <1nim.al test. observation'). The 

cOllfinnatory test I1l..1Y be conducted in a sequential m4UUler using 011e anima l 

per s.tep or using two 3.1lunals. The details of confirm<.1 101Y test will be 

included in the n:po11. 

2. QUALITY ASSliRo\."\TE 

TIle Qlk11ity ASSlmUlce Unit of Advinns Therapeutics Litllired, 

Bangalore - 560 058. India. will UlSp<ct the study. the raw data. the 

draft and final repolt. FUldulgs of all ulSpections will be reported to the 

Management and to the Study Director. ltlSpection dates and repotti..ng 

dates will be entered in the study repot1 . The Quality ASSlu'ancc Unit 

h.as re'viewed the final study plan aud will receive a copy thereof 

3. GLP CO:lIPLJ.o\. "\'CE 

TIle study will be perfolllled in accordmlce with the following GLP 

regulatiom;: 

• OECD Principle. ofGLP (as revised ulI997). ENVIMC/CHEM 

(98)17. OECD. Park 1998. 

• JMAFF GLP Standards for Agricultmal Chemicak ll-Nou5.'nNo. 6283. I October, 1999. 

• EC Directive 2004/IOIEC, Official Jou11lal of the European 

Union, L50!44, Feb. 20. 2004. 

• This study will also be conducted as per the lllUtlk'llly agreed 

study plan and the Stand..'lrd Operating ProcedlU'es, 

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Exceptions to the GLP compliance are listed below: 

• Archiving of raw data at the test £1cility. 

• Exception to guideline 405, for not testing weight of evidence 

analysis by the test facility. 

4. TEST PROCEDUU: GUDELL,,\ES 

TIle prOCedlU"eS as described in this study plan are based on the 

following testing guidelines: 

• OECD Guideline for T esfu12 of Che.lnicals, Section 4 , Number 

405. ''Acute Eye ll.1'itation / (-011'05ion" adopted 24th April 2002. 

• U.S. EPA Health Effects Test Guidelines, OPPTS 870.2400, 

Acute Eye 11l'itatio~ August 1998. 

• JMAFF 12-Nousan-8147. November 2000. 

• Officinl IOlunal of the European COll11Uluuties, Methods for the 

Detennination of Toxicity. Pmt B.5 (Eye In'itation), Directive 

2004!73IEC, 29 Apl'i! 2004. 

5. A.'IEl\lHIE:\T PROCEDUU:S 

All 3mclldulents to and I 01' deviations fiolu this study pIau and the 

reasons therefore shall be documented, d.1ted, signed by ihe Study 

Director, approved by 1ll..:1nagelnent a~ may be required) and 

n1.1intai..ned \vith the study plan. If immediate action is necessary, 

verbal agreeluent \vit.h the sponsor will be cOll.fll"llled as soon as 

possible by study plan amendment., AllY aluendlnems to and/or 

deviations froln this study plan will be recorded ill the raw data and 

doclunemed in the [mal repolt. 

6. SAFETY PRECAT:TIO~S 

Gloves, cap, £,ce n1.1sk and goggles (if required) will be used in 

addition to protective body gal'lucnts and shoes to ensure adequate 

pers on".,1 health and safety und to avoid uilialation and SkUl contact 

\vith the test sub!!.tance. In case of eye contact, the eye will be washed 

thoroughly with \vater and medical treahue11t will be sought. In ca.':.e of 

skin contact~ it will be washed with soap and \:vater with ':.1.1bsequellt 

Iuedieal aid. 

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7. OBJECTIYE 

TIle objective of this acute eye initation <o.mdy in rabbits is to provide 

i..llfoI11k1tion on the potential for ul'itatioll arising fi:om a single 

instillation of a test substance into the eye. 

S. JUSTITICATIO:\ OF SELECTIO:\ OF TEST SYSTEi\I 

The rabbit is the syslem of choice because. historically: it has been the 

prefclTed and most commonly used species for primary eye in'itatian 

tests. 

9. JUSTlFICATIO:\ FOR ROUTE OF AD:'IIL"\l:STR4.TIO:\ 

TIle ocular route of admin.istration will be used because it is 

recollunended ill the referenced. guidelines and because hUllL'1n 

expostll'C 11L.1Y occur via this route. 

10. :'IrA TERIALS A,\'D METHODS 

10.1 "fat.rial, 

Animal 

Source 

No. of rabbits 

Body weight rangeat treatment 

Age at treatment 

Identification 

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New Zealand White R.1bbits 

Inhouse Bred (Out bred) 

New Z<aland White R.1bbits 

Toxicology 

Department ofSafdY Assessment 

Advinus Thenlpeutics Li.mited 

Bangalore 560 058, INDIA 

Up to 3 Male rabbits 

2.2 to 3.2 kg 

YOtlllg: adult (the exact age will be 

mentioned in the report) 

By rabbit accession mUllber. Identification 

of individt!.11 anink11 is by cage label, 

cry~lal violet I tunuel-ic colour body 

marking. 

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Rabbit acces.sion 

numbers: 

Acclimatization 

RB8767 to RB8769 

After physica l examination for good health 

a.nd suitability for experiment. the rabbits 

will be acciuIJ.,tized for at least five days 

before treatment. 

10.1.2 TE'st substance informatioll 

(as fimushed by the Sponsor) 

Test substance ! 

Item 

Lot 1'0. 

Manufactured by 

(Nallle and 

Addre<s) 

Supplied by 

(Name and 

Address) 

Date of 

ll'laIlU£1ctw'e 

Date of expuy 

PlU'ity as per 

Cet1i.ficate of 

Analysis 

Physical 

appearance 

Storage conditions 

(

201000599-6 

The Dow Chemical Company 

Midland. MI 48674 USA 

The Dow Chemical Company 

Midland. MI 48674 USA 

March 2010 

06 March 2012 

99.85% 

Off-white solid 

: Ambient (+18 to +36°C) 

The test ; ubstance. identified as was received 

on 03 Febnk11Y 20 11 and wa s fiuiher identified as 008-26 by test 

facility. Refel'ence Number dated 07 Febnk'lly 20 11 . The test 

substance is a solid and is stored at ambient condition (+18 to +36°C). 

DOClUllentation of the l11ethods of synthesis. £1brication. or derivation 

of the test substance is retauled by the Sponsor. 

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Detenninatioll of the stability. identity. strcngth) purity (lud 

compo')itioll of the test substance is the responsibility of the study 

Sponsor. The follo\ving: illtol1uation related to the characterization of 

the test substance was provided by the sponsor. 

Information as provided by Sponsor are as follows: 

pH : 6.47 (Analysed in-house). 

Specific gravity I density: Not Applicable. Test .mb>tance is a solid. 

10.2 Methods 

10.2.1 H\lSba (h'~' 

Condition') 

Anim"''lis will be housed lUlder stallck1rd laboratolY conditions, air 

conditioned with adequat.e fi'esh a il' supply ( 12-15 ail' clk1nges/houl'). 

EnvirOllluent: temperattu'e 17-2Y'C1 relative InUllidity 30-70 %~ with a 

12 hour light and 12 hom' d .. 1rk cycle. TIle m3xllllulll and lu.iniuIll1U 

temperature and relative hmuidity in tlle eXpe!uuelltal 1'00111 will be 

recorded once daily. TIle relative. hmnidity in the experimental room 

will be calculated :6:0111 dIY and wet bulb temperatw:e recordings, 

Housing 

Housed individually in rabbit cages (approx. size: L 65 x B 65 x 

H 45 cm) with n01yl sh.1llow cage body and £,cilities for pelleted food 

(Stainless steel feed hopper) and drinking water (750 ml mal'holon 

bottle fitted with sippet· tube). The litter collection tray (notyl waste 

tray) will be changed cL1 ily (except on S'Uldays). 

Diet: ad libitum 

Rabbit feed m.allufac.hu'ed by Pranav agro Industries Ltd, Sang li, 

Malk'trashtra .. India , will be provided ad libitllllJ to the rabbi". The 

proclu'ed feed will be subjected to analysis of cOluposition and the 

analysis rep0l1 included in the repot1. 

Specific contaminants (heavy 111etals organochlorines and 

organophosplk1tes) in the diet al'e analysed periodically and the repo1t 

will be included ill the study file. 

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Water: ad libitum 

Deep bore-well water passed through activated charcoal filter and 

exposed to UV rays in Aquaguard in-line water filter-cum-plU'ifier 

manufactured by Elu'eka Forbes Ltd .. Mlunbai - 400 001. INDIA, will 

be provided. TIle water bottle will be replenished once daily and the 

water bottles changed once a week. The water alwly, is resnlts will be 

fiunished in the repol1. 

10.2.2 Test substance instillation 

Prior to instillation, both eyes of each rabbit will be examined with the 

aid of a pen light for in'itation and ocular defects. TIle eyes will then 

be checked for opacity using ophtlk11mic fluorescein sodium test strips. 

The eyes will then be irrigated v .. ith enough physiological saline 

(0.9% NaCl) to remove any excess fluorescein (approxink1tely 

30 seconds). Using an ultraviolet lalnp. the eyes will be checked alld 

evaluated for pre-existing comeal ci1nk1ge. Rabbits showing any 

defects will be eliminated and replaced. 

For solid test snbstance: 0.1 g of the finely grolUld test item will be 

placed in the everted lower lid (conjlUlctival sac) of the left eye of each 

rabbit. TIle lids will be gently held together for about Olle second. in 

order to minimize loss of the test substallce. 

If it is knowll tbt the test substance may cause comeal opacity or pain 

il1'itatiolL or if pain or discomfo11 is noted after any anuml is instilled, 

the procedures for lese of anaesthetic will be followed as described Ul 

Appendix 1. 

A.'1L'1IAL SELECTION 

Stepwise starting with a single animal: Selected 

10.2.3 Animall'esfminf 

TIle Elizabethall collar will be fixed around the neck of rabbits and 

retauled until the eye score, if any 1'enU11 to zero 01' till 72 holU's, 

whichever is earlier. 

10.2.4 Stel",ise exposure of animals 

A single rabbit will initially be exposed to the test substance. 

Prior to instillation of the fest substance, a local anaesthetic nwy be 

used in both the test and control eye. If a cOll"Osive effect is observed at 

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fUlY scoring interval. the te.<.t will be tenninated llllluediatdy and no 

fUl1her testul£! will be conducted. If no conusive effect is observed <lnd 

inltation doe'S not s~t tluollgh Day 21. the test will be completed 

using two additional an.im<1 Is. 

10 . .3 OIJSE'lTatiollS 

10.3.1 Clinica l obs("l'Yatiolls 

Rnbbits will be observed fOl' toxic signs and pre-te.muual deaths fotu' 

times (at hotll'ly inrervab,) 011 the d.1.y of test substance im.tillation and 

once daily thereaftet' till the end of observation. Observations will 

include gro.'.s evahla lion of skin and fi.u·) eyes and mucous. membranec... 

re.spiratolY. ci.rcu1.1tory, autonom.ic and central nervollS systenls. 

5.0matOlllotor activity aud behavior pattent. 

Palticular attention will be directed to observation of tremors. 

convulsions; salivation. dinn'hea and COllla. 

10.3.2 Body weights 

TIle body weights will be recorded at the 5ota11 of accluuatization. d'\y 1 

of treatment Gust before test substance ul50tillation) and at te.J"Illiuation 

of observation/death. 

10.3.3 Eye s('ol'ing 

OcuL."1f le oll~ will be evaluated with the illumiuntion of a white light 

sO\U'ce at interval5 of approxim ... '\tely 1. 24, 48 and 72 hom's postinstillat ioLl of the test substance according to the "Scale for Scoring 

Ocub,. LeSiOllS'· (DRAIZE. 1977). 

1. co',.n;:-;CTIYAE 

SCORES 

(A) Redue'is (refe-r to palpebral .,nd bulbar coujutlclI\'ae) . 

Ve-ssels JlofllL'"\l .. . ... . ..... ......... ... ... ... ..... .................. 0 

Vesse ls definitely inj ected abO\'e norm,,'! ... . 

More diffu'ie, deeper criJus:ou red. individual vess:els 

not easily di'>cernible . ................ .. .. .. ... ....... .. . 2 

Diffuse beefY red ... ............ 3 

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(B) C ht>Inosis 

Nornm} ................ . 

Any !;wdhng above nOIul<1.1 (including nictitating: 

tue1ll.bnule) . 

o 

ObviollS s'welliug \ ... illl pat'tial e.,' ersion of lids ... ... 2 

Swelling with lids abom h alf closed .. . ........ 3 

Swelling with lids half closed to completely closed 4 

(C) Di'i l" harge. 

No d:i-.:charge . . .. .... ............................................. 0 

Any amouut diff~ent from norm.al (does not include 

5111<\ 11 amollnts obseryed In ilUler canthu.s of llOfllli'll 

alum .. ,ls) . . ............................................... . 

Di':>charge- w ith moi'> ' t!'lUl1g of the- lids and hair<; j U5t 

ndjncenr to lid, .................. .. .. . .. ........ ... .. 2 

Discharge with Uloi'> tening of the lids and h<1.lc.c:.: a 

Considerable area around the eye ..................... .. ......... ....... 3 

TOTAL SCORE ~( B +C) ' 2 ~L·\XL\IV?I TOTAL ~ 20 

2. IRIS 

(D) 

Folds above norma l. congestion. swelling, 

Circulllconlc-al i.llj~tion (any oue or all of the<'je or 

C'ombiu.1tiotl of allY thereot) iris still reac ting to light 

(sluggish reaction is positi"'e) ................................. . 

No reaction to light. ha emorrhage, gr oss desttuction 

(anyone- or all of these) .............. . 

SCORES 

o 

2 

TOTAL SCORE ~ D , 5 1\L-\....x:nIl1\I TOTAL = 10 

3. COR.'\EA 

(£.) D egn..e- of Op:lcit~· (most dense area used) 

No opacity 

ScatTered or diffue;e areat;- detaile; of iris clearly 

nsible 

Easily disceruible translucent areas. details of iris 

SCORES 

o 

Clearly vi')ible . ....... .......... ..... . ...... . ... ... ... ...... ..... 2 

Opale')cem area'). no details of iris ,,·isible. size of 

pupil bardy di .. cernible .... ................ 3 

Opaque. iris mvit;ibJe ... . .......... ...... 4 

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(F) AI'(>:1 of C:Ol'ut':l !U\"oIYE'll 

One quartet" (or Ie':> !» but not Ze1"O . . ... . ........ , _ ............ . ......... . 

Greater than one quartet" but less than half 

Greater th.1n half but le<;5 than three quarte1"<; 

Greater tl1..1n three quarter .... up to \"hole nrea 

THE TOTAL SCORE = (E' F) x ;. 

2 

3 

4 

)L-LTI\Il~1 TOTAL = SO 

)L-L'U)Il~1 TOTAL SCORE POSSIDLE = 110 

In addition, all treated eyes will be eX.1lnined using ophthal.mic 

fluorescein sodil1ll1 at the 24-hotu' interval. If cOluea} opacity exists at 

24 h01U'& ~ fluore scein \vill be llsed, as needed.. at subsequent scoring 

intervals to aid in eVahk'l.ting: t.he extent of c01l1cal d-11uage. 

If there is no evidence of ocular in'itation in any test rabbit at 72 hom's. 

the study will lx tenllinated. If irritation pert:.ists in any rabbit at 72 

hom'" scoring will continue fol' all rabbits at 4, 7, 10, 14, 17 and 21 

days post-instillation or \uttil all signs of in'itation ru'e completely 

reversed (n score of zero) in all rabbits. whichever OCC1U'S first. The 

dlll-arion of the scoring period will not exceed 21 days unless 

authorized by the SPOllSOL 

Individual scores will be recorded for each rabbit, All score; will be 

calculated as described in the "Scale for Scoring Oculru' Lesions". ru1(\ 

the totnl possible points (out of 110) will be tallied, In addition to the 

ob.,ervation of the cornea. iris and conjunctivae, any other lesions that 

al'e observed will be noted, To aid in the inte1pretation of data, the 

average score for all rabbits at each scoring period will be calculated, 

If, at any tune dtu-ing the study, extensive in-itation is noted and 

deemed in'eversible, the Study Director will notify the Sponsor thM the 

study should be tenuinated. If dallk'lge is extensive and the Sponsor is 

lUlAvailable to authorize termination of the study. the Study Director 

may unilaterally decide to have the animal> euthanized inullediately 

for hUll1.1Ue reasous and the Sponsor will be notified as SOOl1 as 

po",ible, 

11. RESL"LTS 

11.1 Data compilation 

The fo llowing ck,ta will be recorded on ck,ta sheets and transcribed for 

COlllpilation and analysis: description of the degree and nature of 

in·itatioll. the pl'ec,ence of ocular lesions. and any effects other than 

oculru' noted during clinical observations and body weights, All 

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fmding; will be recorded m the stalldru'd repolting fonuMs and 

presenred in the repolt. 

12. REPORTL,G 

The fmal repolt will include the following infonnation: 

}- The name: and addrc:o;.s of the spoll.'!.or and the test facility [liong 

with the study schedule. 

:,... The llames and signattm!s of all persolU1el i.nvolved in the S.tudYl 

including the Study Director ruld other scientists. 

"; TIle test 5.ub')tance with identification either by name and I or code. 

~ PlUlty. composition and other appropr;'1te chru'acteristics of the test 

subs.tance. 

,. Test <;.ubstance : 

• Test substance inf0I111ation as prov-ided by the Sponsor 

, Vehicle: 

• Identification. concentration (where appropriate), voltulle u')ed. 

~ Test anima ls: 

• Specieslstmin used 

• Age of allinlal., at strut of treanllent 

• Ntl111ber of animals and sex 

• Individual animal -weights at strut and cOllclus.ioll of test 

• SOllrce. housing: conditions, diet, etc 

, Results: 

• Description of method llsed to score inilfltion at each 

observation time 

;,.. A ~UnUll.c11y and description of all results.. 

,. All study plan deviations. if any. 

,. Individual toxic signs and gro.s.s necropsy fUlding~~ if any. 

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~ A description of all ci:rcmllstancec; if any, that lllay have affected 

the quality or integt'ity ofrhe study. 

»- Quality A'isurance Statement. s.igned by the Quality A'iSUrallCe 

Unit. 

» GLP Compliance Statement. signed by the Study Director. 

~ The sTOrage location of all raw dat.a, the report. a sample of test 

substance and the archiving period. 

;;.. GLP Cetiificates. 

TIle original signed final report will be sent to the Sponsor and one 

signed fmal repo11 will be retained by Advinlls. The original raw data, 

docwnentation, records, the original study plan, and final repol1 

generated rI'3 a 1'e501t of this study \vill be sent to the Sponsor for 

retention follo\ving signing of the fUlal repOlt. Copies of the saIne 

(authorized by the Study Director) will be retained by Advmus. 

Aclvinns will maintain th~~~ r~cords for a period of at least five y~ars _ 

After tills time, the Sponsor of the study will be offered the opportlUllty 

to take possession of the records or will be charged an archiving fee for 

continued ru-duving by Advi.lltls or to dispose of the raw data in 

con'S:l1ltation \vith the Sponsor_ A representative salllple of 

approxlluateiyone gt-anl of th~ test itenl will be ~nt :£i:om the test irenl 

stores to the archives in the fest £1cility before the fu-st dispensing: of 

test item to studies. Tills sample shall be stored for a period of 2 years 

fi'om the date of rllls fmal ropo1t or till next GLP inspectiolL wlllchever 

is later. however nol beyond 5 years_ 

In the event of any lllsp~ction by regulatOlY auth.oriti~ !.; seeking 

original raw data for verificati ~ the sponsor \vould be re spo n~ible 

either to seud the origina l raw data to Advinu5 Therapeutics Li.mited or 

to the regulatOlY ag\'!"llcies for verification_ 

14. STL'DY PLA .. " DISTRIBrnOl\' 

TIle f11101 study plan (original copies) will be distributed as follows: 

Sponsor: Soft copy of the sigtled Study piau in PDF format and one 

signed study plan ill original (Copy No. 111). 

Study Director: One sigtled study plan in original (Copy No. 111). 

SP- EYE 

Copy o_ 111 

. EYE 

Copy No. 2/2 

Pilge 15/19 

Page 38/50 
- 

APPENDIX 3 contd. Study Plan 

15. REPORT DIsTruBl-no:" 

n," fUlal repOlt (original copi,,) will be distributed as follow;: 

Test Facility Archives: Olle 'iigned fUlal repol1 in original 

(Copy No. 112). 

SpOJlsor: Oue signed fmal repolt in original (Copy Kos.2/2) and one 

Soft copy of the r<polt in PDF fonnat. 

16. REFEREl\CES 

1. Draize. J.H., Woodward, G and Calvely. H.O 1944. 

J.Phal111acol. Exp.TIler.83: 384. 

2. Draize. J.H 1977. Dennal Toxicity In: Appraisal of the sa fety of 

chemicals in Foods. Dn.lgs and Cosmetics. Association of Food 

and Dmg Officials of the United States. Austin, Tex.1s. p. 47. 

3. OECD. 2000. Guidance Document on the Recognition. Assessment 

and Us.e of Clinic.:'ll Signs as HlIluane Endpoints, for Experimenta l 

Anim •. 1.1s Used in Safety Evaluation Envu'olllucntal Health and 

Safety Monograph Series 011 Testing and Assessment No 19. 

(ENV/JMiMONO (2000) 7. 

SP- EYE 

Copy:'\o. 111 

EYE 

Copy No. 212 

Pag(' 16119 

Page 39/50 
- 

APPENDIX 3 contd. Study Plan 

17. AGREEMENT 

This Study Plan for Sludy No.:07694, "Benzoylated Calechol: Acutc 

eye irritalion study in Rabbits" has been mutually agreed and sjgned. 

Sponso r Study plan Approval: 

nlC (inal version or the study plan wa::; approved by cJnail frolll the 

study monitor on 07 March 2011 for Study Director's Signature. 

ror TEST FACILITY 

STUDY DlRECTOR 

Da!e: 7'" ,,,,-,,., 2<> I, 

QUALITY ASSURANCE UNIT 

Da{c:Qe ,mnrnf1 , )~" 

MA TAGEMENT 

Date: () ~tI\«d.V' I 

SP-G769",_EVE 

Copy No. III 

EYE 

Copy No. 212 

I'Rg(' 17/19 

Page 40/50 
- 

APPENDIX 3 contd. Study Plan 

APPL\lHX 1. PROCEDl."RES FOR USE OF .<L'\ESTHETICS 

DVRl;,;G OCTLAR IRRITATIOX TEST Il'\ RABBITS 

C\ot~ : The- pl'o(eclul'£>s des cribed are gE'u(>ral iu U:ItUI'!;' :md only those 

pl"oC"E'rhu'es applicable based au the u;'lture/C'h:u';lcteristics of the test 

substance will be followed) 

Pre-Application 

If the pH or formulation of the test ~ub tallce is sUlular to other test 

compound, (previou,ly tested) that h.we produced comeal opacity or 

pain uTitatiOlL 2·3 drops of Lidocaine 4 % will be ulStilled Ulto both 

the treated and control eye prior to dosing. Dow will provide the 

infol1uatioll regarding the previollsly h: ted material. 

Post-application 

Lidocaine 4 % will be. initiated inunediateiy after dosing if exces')ive 

pawlllg: at the eye OCCllI'S (in the event of powder administration. the 

physical properties of the powder may cause transient pawing, there ~ 

not a need to treat in this case) or if pain response is more significant -

excessive vocalization or trying to escape. Two to three drops of 

Lidocaule 4 % will be ulStilied Ulto both the treated and control eye. 

Que hour Examination 

No TreatnlS:ut: If there is no or slight conleat opacity ($cores of 0, 1), if 

there is slight conjunctival redness (scores of 1. 2). slight cOl~ullctival 

swellulg (scores of L 2). discbarge (scores of L 2) and u·itis (scores of 

O. 1). The rabbits will not be treated with an.1esthetic or analgesic. 

OR 

Initiation oftreatmellt with Lidocaine 4% anaesthetic (evelY 2 holU'S): 

Treatment with Lidocaine 4% is initiated if the rabbit iO) observed with 

significant cOllleal opacity (scores of 2, 3. 4)1 cOluunctival swelling 

(scores of 3. 4). di, charge (score of 3), conjunctival redness (score 

of 3) and iritis (score of 2). Two drops of Lidocaule 4 % will be 

ulstilled evelY two homs, tJu-ougbout the workdny. to both the t",ated 

and control eye. 

R.1bbit Re·e\·alllated at the Eud of Da~· 0 if Lidocaine oj % 

treatment has be-en initiate-d 

Tennin"tion of Treatment: If there is no or slight comeal opacity 

(scores of O. 1). if there is slight conjunctival redness (scores 1, 2), 

slight conjunctival swelling (scores of 1: 2) or discharge in the absence 

of sigllificnnt fUldulgs (score, of 1, 2). 

SP· EYE 

Cop)' Xo. I II 

EYE 

Copy No. 212 

P.,gE' 18.119 

Page 41/50 
- 

APPENDIX 3 contd. Study Plan 

OR 

Initiate sy')temic treatment: If ocular in1.tation has wOl'<:.ened or 

remained the same from the One Hotu· E valtL:1tiollo; admin.ister 

BuprellOIphine (0.01 to 0.03 mgikg by IVIIMlSC) at the beginning and 

elld of the workday. 

RE'-fTaluatioJl of rabbit dUl'ing thE' first 72 hours post closeInitiate systemic treatment: Treatment \\o"ith Buprenorphiue at the 

beginning and end of the work day (approximately 10-1 2 hour 

interval). is initi.,ted if the rabbit is observed with significant comeal 

op acity (;cores of 2. 3. 4). conjunctival swelling (scores of 3. 4). 

coqjunctival redne>s (score of 3) and iritis (score of 2). Two to three 

drops of Lidocaine 4 % nk1y also be used prior to staining of the eye 

\vith fluorescein, and rinsing. to alleviate any potential discomf0l1 that 

tills process (spec ifically ti-onl the 111echmucal aspect of rinsing. the 

eye) nl<'y cause the rabbit. 

TI,e topical anaesthetic will be administered at least 30 seconds prior to 

the usc of the fluore scein. 

TE'l'lnillatioll of S~·stemic Treatment 

Treahllellt lllay be stopped when conjunctival uritation ~ comparable 

to the olle-hotu' evaluation criteria for not. treating dIe eye with 

Lidocaine 4 %. and no conleal defects are observed (greater tban a 

score of 1) after staining with fluoresceul. If at any time. the aninlal 

appears to be in undue pain/distress (as indicated by clinical 

observations confU"llled by a Study Director arullor Study veterinarian): 

the an;'1I<,1 will be euuUUlized and it will be noted in the study records. 

SP-Gi 694_EYE 

Copy::'\o. I II 

EYE 

Copy No. 212 

Pnge 19/19 

Page 42/50 
- 

APPENDIX 4. Deviation from the Approved Study Pilln 

Study No. 

Study Title Acute Eye Irritation Study in 

Rabbits 

Test Item Code D08-26 

SI. AS IN STUDY PLAN DESCRIPTION Ol? DEVIATION No. 

I . Pages 4/19 and 5/19 

1. STUDY DETAILS STUDY DETAILS 

Stud y Sched ule: Study Schedu le: 

Draft report: Latest by 30 April 201 1 Draft report: 25 May 2011 

Study completion date: Latest by Study compl etion date Latest by 

I May 20 11 30 June 20 II - -------,-- Explanation : To in clude the change 111 draft report sublllission date and st udy 

com leti on date . ----------- 2. Page 13/19 

10.3.3 Eye Scoring Eye Scoring 

In addition, all treated eyes will be In addition, the eye of one rabb it which 

examined lIsing ophtha lmic fluorescein showed cornea l opacity at 24 hours was 

sodium at the 24-hou r interva l. If cornea l exam i ned lIsing ophthalmic flu oresce in 

opacity ex ists at 24 hou rs, fluore sce in wi ll sod ium at the 24 and 48 hour interval. At 

be used, as needed, at subsequent sco rin g 72 hours observatio n, th is rabbit had no 

in te rvals to a id in eva luating the extent of opac ity and no eye sco res and he nce 

cornea l damage. fluorescein was not used and terminated 

the ex periment. 

Other two rabbits where there was 110 

cornea l invo lvement ,-vere 110t obse rved 

usin g ophtha lm ic nuorescein. 

Explanation : To correct and include the rabb it eye examination using ophthalmi c 

fl uorescein. 

Date: I b 'It) "-' 2.0 t( 

EYE 

Copy No_ 112 

(P. J. I)rakash) 

Study Director 

Page 43/50 
- 

EYE 

Copy No. 2/2 

15. ANNEXURES 

Page 44/50 
- 

ANNEXURE 1. Analysis Report - Animal Diet Sample 

ADVINUS TITERAI'EUTICS LIMITED 

21 & 22, P EENYA INDUSTIUAL AliliA, IlI'IIASE 

I3ANGALORE . 560 058 

ANALYSIS n El'OnT (AM EI'OJ<:O)· ANIMAL IlIET SA M PLE 

From: Ana lytical R & D Department 

Advinus, Danwtl ore-560 058 

To: Department of Safety Assessment 

Advinus, B:ulgalore-560 058 

Our Ref. Nq. : WCn T S12 14 7 Date: 14. 06.20 11 

Sample Details: Name : Rabbit Fct'd Sampling Date : 19.10.20 10 

Balch No. : 05 9~ 

Supplier : Prannv Agro Industries Ltd., Bllngalorc . 

,Manufaclurer Pralluv Agro Industries LI'd .• Slll1 gli. i\1ahmashtra. India 

SI. No. Paramete r 

ANALYSIS RESULTS 

(An:llysis on " ;IS i!i basis") 

('/oj 

---I~-- -M ;;'~ --- -- -------------------------------- ----- iT

-'--'i:-- -C ~d ;;;-cN 6.25 :;'-----------------------------------------------20.32·· 

3, Crude fal (E ;' xtrac r-------------'- -- ----' ---"-'--" -'--' ''' '' '-'- ---'j'~i -"" 

-'''- 4~''''-C~ fi b;'~-- ''-'-''''---'' --'----''--'-'' ·······--···--·-----···· .. -·--.. ·· --·--·- ~98 .-

. ···-- ---··· To l~ j"I ·-···-···-· .. ·-·-·-··--·---··-·-··--···-··-·····-··--···-·-··--· .. ··-· ·· ··- -·······-- ~3)-" 

6. Acid insoluble fi sh --- .... _ ..... -.-- -'-" ..... ' 43"2 -' 

8. Cn lciulll (en) 0.54 

Rt!ason for Amendment: To co ~c l the Manufacturer's address. 

·EYE 

Copy No. 2/2 

Analytical R&D Ocpat"tmen t 

Page 45/50 
- 

ANNEXURE 2. Analysis Report - Water Sample 

A D V I NUS TH EH A PEUTICS LIMITED 

21 & 22, PE"ENYA [ NDlJSTRIAL AR EA, II P I-l ASE 

BANGALORE 560 058 

ANALYSIS HEPORT· WATER SA MPLE 

From: Analytical R&D Department 

Advinus, Bangalore-560 058 

To: Depm1menl of Safety Assessment. 

Advin us, Bangalore-560 058 

Our Ref. No: WCrrWS1257 Dale: 24/03/201 1 

Sample Details: Source of Collection: Outlet of the Aquaguard (At use point) 

Date of Collection: 1110212011 

ANALYSIS RESULTS 

51. Parl'lmctc." 

No. 

I. Colour 

2. Odour 

3. Turbidity 

4. PH 

5. Electrical Conductivity, 

dSm-' 

6. Total solids, (ppm) 

7. Suspended solids, (ppm) 

8. Dissolved solids. (ppm) 

9. Dissolved oxygen (ppm) 

10. Biochemical Oxygen 

Demand 5 days a! 20°C. 

(ppm) . 

II . Chemical Oxygen 

Demand (PPn'I) 

EYE 

Copy No. 212 

COli t-en I 

Colourless 

OdourJess 

Clear 

7.36 

1.324 

963 

18 

946 

4.9 

2.1 

6.2 

SI. 

No. 

12. 

13. 

14. 

15. 

16. 

17. 

18 

19. 

20. 

Parameter Con tcnt 

m 

Total hardness as 220 

CaC03 

Calcium as Ca- ~ 37 

Magnesium as Mg-- 31 

Chlorides us CI" 283 

Sulphates as SO~ - 50 

Carbonates as C03-· Nil 

Bicarbonates as HC03· 290 

Sodium as Nil. 24.8 

Potassium as K 2.79 

Atwlytical R&D Depat1mcnt 

Page 46/50 
- 

ANNEXURE 3. GLP Certificate - Germany 

I 

I' 

I " 

Bundesinslilut fOr Risikobewertung I 

Federallnstitutc for Risk Assessment FlcBfR 

GUTE LABORPRAXI S I 

GOOD LABORATORY PRACTI CE 

(gernM I according to § 1 Ob Alls.2 Nr.3 Chernikaliengesetz I German Chemicals Act) 

bne 3 LP.lnspektion wurde durcl,geftihrl in , A GlP inspecfio,; was earned out at 

Prilfeinrichtung I Test Facility 

Advinus Therilp" IhcS Limitert 

(former name until October 201 0; Advinus Therapeutics Private Limited) 

21 & 22 Phaso II, Pc-snya Industrial Area 

Bangalore - 560 0:';8, INDIA 

PrOfkategoricn I Area of Expertise 

PrOfungen zur Bestimmung dcr physikarisch-chemischen Eigenscl1aften und 

GehaUsbestimmungen I Physical-dlemicallesting 

Pliifungen zur Bestimmung der toxikologischen Eigenschaflen I Toxicity studies 

PrUfungen zur Bestimmung erbgutver~ndernden Eigenschaften (if) vilm, in vivO) I 

Mutagenicity studies 

OkCIOxikologische PrQfungen lur Bestimmung dar Auswilk' !(lgen auf aqualische 

und lerrestrische Organismen I Environmental toxicity studies on aquatic and 

terrestrialorganismr: 

Pruflln!)en Zlim Verhallen im Ooden. im WSSEor lind in dcr Luft: I3lookkumulolion I 

Slud o.:~ 0 11 behaviour tn soit, water and air: bioaccumulallon 

PrOfungen zur Bestimmung von RO(;kstanden I Residue studies 

Analytische PrUfungen an biologischen Materialien I Analyllcal and clinical 

chemistry testing 

Sonstige PrOfungen - Blonanalytik Pharmakokinetik, Sicherheitspharmakologie I 

other studies - bioan<llytics. pharmacokinetics and safety pharm<lcology 

Datum d er Inspektion I Cate of Inspection 

July 07 - 10, 2009 

Auf der Grundlage des lnspeklionsberichtes und der Besprechung Uber zu erfolgende 

MaBnahmen wird hiermit bosUnigt, dass in dieser PrOfeinrichtung die oben genannten 

PrOfungen unter Einhaltung der GLP·Grunds.:ltze durchgefOhrt werden k6nnen. I 

Based on the inspection report and the discussion of follow up activities it can be 

confirmed, that the test facility is able 10 conduct the aforementioned sludies in 

compliance with the Principles of GLP. 

Eine Oberprlilung dieser GLP-Beslatigung ist spateslens drei Jahre nach der o. g. 

lnspektion 7.U beantragen. Dime diesen Antrag wird nach Ablauf der Frist die 

PrQfeinrichlung aus dem dculschcn GLP·Obcrwachungsprogramm gcnomme/l und dicse 

GLP-Best.:ltigung verliert ihre GOltigkcil. I 

Verification of this GLP Certificate has 10 be applied three years after the above 

mentioned inspection al the latest. Elapsing this term. the test facility wilt be taken out of 

the German GlP Monitoring Programme and this GlP Certificate becomes invalid. 

" 

I 

~ I 

'I 

, 

" I 

i 

I 

i 

J 

" 

1 

I 

EYE 

Copy No. 2/2 

Page 47/50 
- 

ANNEXURE 4. GLP Certificate - The Netherlands 

vocd f)1 e" wa ren autoriteit 

ENDORSEMENT OF COMPLIANCE 

WITH THE O!;;CD PRINCIPLES OF 

GOOD LABORATORY PRACTICE 

Pursuant to the Netherlands GLP Compliance Monitoring Programme and 

according to Directive 2004/9/EC the conformity WiUl the OEeD Principles of GLP 

was assessed on 4 - 7 May 2009 at 

Advinus Therapeutics Private Limited 

21 & 22 Phase II , Peenya Industrial Area 

Bangalore - 560 058 INDIA 

It is herewith confirmed Ihat the afore-mentioned test facility is currently operating 

in compliance with the OECD Principles of Good Laboratory Practice in the 

fOilowing areas of exper1ise: physica l·chemical testing, toxicity studies, mutagenicity 

studies, environmental studies on aquatic and terrestrial animals, and analytical ancl 

clinical chemistry. 

EYE 

Copy No. 212 

4'/~:(i'~L~' :':"~~ : ':. 

1~~.-{~;{,:« +h~\"agUe. 27 August 2009 

~i~;i -...,.:.:;.~, .. ~_ .. :~ :..:,. 

Manager GLP Compliance MOflitoring Program 

Food d Con::>ume r Prcduc1 Silfefy Authority (VWA) 

Prlnses Beatrj),:Jagn 2. 2595 AL Den Haag 

Pcstbus 19506, 2500 eM Den Haag. The Netheriands 

Page 48/50 
- 

ANNEXURE 4 contd. GLP Certificate - The Netherlands 

Food and Consumer Produce Sarety 

AUlhorit) 

Mil!isrry 01 A9ricu/rure, Nacure and 

Food QualilY 

Advinus Theri"!peutlCs Limited 

for the attentio" of Dr. S. Natesan 

Vice President21 & 22, Phase II, Peenya Industrial Area 

Bangalore • SGO 058 

INDIA 

Date 18 November 2010 

Re. Change of name 

Dear Dr. Natesan, 

I have received YOllr letter of 26 October 2010 in good order. Tn yOllr letter you 

explained that Advinus Therapeutics Private limited has changed its name into 

Advinus Therapeutics limited. Further, you stated that the change of name has no 

consequences fo,' the ownership and that the test facility will continue to function 

as earlier. 

The GlP Compliance Monitoring Authority of The Netherlands has periodically 

inspected Advinus Therapeutics Private Limited. The most recent inspection was 

carried out on 4 - 7 May 2009. Subsequently, the inspectorate issued an 

Endorsement of GLP Compliance on 27 August 2009. This endorsement reflects 

the name of the test facility at the time of the inspection and will not be renewed 

until a new inspection has been conducted. 

Yours sincerely, 

Manager GLP Compliance Monitoring Program 

EYE 

Copy No. 212 

Pn"~('s ~Jvi KIJall 2 

2!J'l) ilL Olin Ha~9 

1'0 !lex 1951}6 

25 i1 eM D~n HMg 

The Netheri~nd5 

ww-" vwa.nl 

Conl;lct 

Or. R..M.II. JaSpe'S 

T +31 {OPO 448 4769 

M "310 

f· dl {O)70 449 ~747 

r Cb j.JSp("~(~vw~.nl 

Our ref. 

TR.O ... ·.WnI0!293l1 

Page 49/50 
- 

ANNEXURE 5. GLP Certificate - India 

NATIO·NAL GLP COMPLIANCE l\fONITORING AUTHORITY 

GOOD IAHOIlATORV PRACf1CE 

GLP CERTIFICATE 

GLP Inspection was cmricd out a1 'Ad\'inus Thcr:~pcutics P,"ivatc Limited, Plot Nos. 2 t & 

22, Phase-II , Post Dox No. :;813, J'ccnya Industri:d Area, Bangalor..c-S60 058, Inclj" in the 

following areas or c;.;pcrt ~c : 

• physical-chemical testing 

• toxicity studi es 

• ntu(;lgcnicit), studies 

• cn\'irol1mcntalloxic.ity studies on aquatic & terrcs.tri!ll organisms 

• studies on behaviour in water, soil and air, biollCl"Umu lation 

• ,'csiduc studies 

• ;\na"fi.rtical and clinical chcmisll1' testing 

J3:ls~d Oil lhe Inspection Report and the lollow-up actions taken by the tcst ftlci lity. it is 

confirmed that the test faci lity is.capahlc of CClIlducling the ahove·mcntioncd tcstsistud i(:s in 

compliance wi th OI CD Pdllciplcs of Good Laboratory Practice (GLP) :lnd Norms, as 

adoptcd by the Na ti {) ll GLP COlllplinllce Monitoring J\uthorit y. 

This GLP C~11i ficatc is valid fbI' a period of three vea l's frolll Novembe r 7, 2008 subject 10 Ih\.' 

condition that (he lest fa cility compl ies with (h e Terms & Conditions of Ihe Na tion:,1 (~LP 

Compliance Monitoring Aut horj ty's Document Number GLP-I01 . 

Ccrtificate No.: GLP/C-OI9 

Issue Datc : 

·EYE 

Copy No. 2/2 

24·12-2008 

Head 

Nation:)1 GLP Compli:Htce Moltilol'ing Authority 

Depa rtment of Science & ;rechnology 

Technolo~ Hlta".m New Delhi-I 10016 

Page SO/SO