Document zbbV31odNvpzedvoYLjQdKRJ3

rn K J AR226-2993 Baugy, June 17, 1993 STUDY No_9200BA c '1 OCULAR PRIMARY IRRITATION Sponsor : ELF ATOCHEM SA La Dfense 10 Cedex 42 92091 PARIS LA DEFENSE FRANCE Company Sanitized. Does no! contain t c /* i 'C: C iS :C CS ' TCLcS 'CCIC F - ftLeCOPE :1S> -k/' CC k ' CERB - STUDY No. 920084 E TRIAL COMPOUND : Ocular primary irritation. SPONSOR SPONSOR CONTACT ADDRESS SITE OF TRIAI : ELF ATOCHEM SA : Mr REGNIER : La Dfense 10 Cdex 42 92091 PARIS LA DEFENSE - FRANCE : CENTRE DE RECHERCHES BIOLOGIQUES Chemin de Montifault 18800 BAUGY, FRANCE RESPONSIBLE STAFF FOR CFRR : Scientific and Technical Director : S. RICHARD Pharmacien Docteur de 3me Cycle Matre en Pharmacologie Date Signature Head of Toxicology Department : C. AUDEVAL-GERARD Docteur Vtrinaire CES d'Ophtalmologie Study Director : C.-BESSON D. U.T. de Biologie Applique Quality Assurance : C. VIGIER Date Signature Signatur`Oe The present report is based upon the actual and authentic experimental results obtained. &an tt contaiiTSCACBI CERB - STUDY No. 920084 E This report consists of 35 pages numbered from 1 to 35 including 5 appendices. CONTENTS I. SUMMARY Page 4 II. EXPERIMENTAL PROTOCOL ADOPTED 11.1 INTRODUCTION 11.2 AIM 11.3 METHOD 11.4 COMPOUND 11.5 ANIMALS 11.6 ADMINISTRATION OF COMPOUND 11.7 CLINICAL MONITORING OF ANIMALS 11.8 CALCULATION OF OCULARIRRITATION INDICES 11.9 STEPS OF THE STUDY 11.10 CLASSIFICATION 11.11 DATA ENTRY OF RESULTS 11.12 REPORT 11.13 QUALITY ASSURANCE 11.14 RECORDS III. RESULTS HI-1 111.2 111.3 DEVIATION FROM PROTOCOL No 92.04.10.03 DATES OF TRIAL OCULAR TOLERANCE IV. CONCLUSION APPENDIX 1 : Protocol No 92.04.10.03 APPENDIX 2 : Technical "data and analytical certificate concerning compound APPENDIX 3 : Analytical certificate concerning foodstuff APPENDIX 4: Analytical certificate concerning water APPENDIX 5: Results data sheets 5 6 6 6 7 7 8 8 10 11 11 11 12 12 12 13 14 14 14 17 18-25 ' 26-28 29-30 31-32 33-35 [|jj?mpany Sanitized. Does not contain TSCA CBI CERB - STUDY No. 920084 E I. SUMMARY The aim of this study was objective determination of the irritant properties and/or degree of corrosion induced by c o m p o u n d w i n g single instillation into the eye of the Rabbit. ** ^ 0.1 ml of c o m p o u n c jjH H H lI f lf e f w e r e instilled into the conjunctival cul de sac of the left eye o fth e three rabbits used. The non-treated right eye served as a control. One hour, then 24 hours, 48 hours, 72 hours and 8 days after instillation of the compound, any possible conjunctival, iris and corneal lesions were evaluated. RESULTS OBTAINED WERE AS FOLLOWS : Mean indices were calculated from results obtained in each rabbit at times 24 hours, 48 hours and 72 hours. - Cumulative means of mean indices were : . Chemosis: 0.78. . Redness : 0.89. . Iris : 0.22. . Cornea : 0.00. - Mean indices per rabbit, for the three times taken together were : . Chemosis: 1.00 in one animal and 0.67 in tw o animals. . Redness : 1.00 in tw o animals and 0.67 in one animal. Iris : 0.33 in tw o animals and 0.00 in one animal. . Cornea : 0.00 in all animals. In conclusion, compouncj| /as found to be non-irritant for the e y ^ jH h e Rabbit. fafiy'Sanllfeatf. Doestrot contain TSCA CB* CERB - STUDY No. 92008 4 E D II. EXPERIMENTAL PROTOCOL ADOPTED CERB - STUDY No. 920084 E 11.1 INTRODUCTION 0 The protocol used for study of ocular primary irritation was in accordance with the requirements of O.E.C.D. Guideline No. 405 (February 24, 1987) and E.E.C. Directives (67/548, 79/8311, 83/467, 84/449) and enabled full evaluation of the reversible or irreversible nature of effects seen. II.2 AIM The aim of the study was objective determination of the irritant properties and/or degree of corrosion induced by c o r n p o u n d f l M m ^ m H H H H H ^ f m P | | B n 0^0 w 'n9 single ocular instillation in the eye of the Rabbit. * II-3 METHOD Ocular irritation and/or corrosion following instillation of compound] f l D i n the conjunctival cul de sac of the left eye of the Rabbit were evaluated by grading of the ocular irritation reactions seen, using a predetermined score system. This method was not applicable to substances w ith a pH of less than 2 or more than 11.5, nor those found to have a definite corrosive action or to be source of severe irritation during dermal irritation tests, for which a corrosive effect on the ocular mucosa is then predictable. The protocol n 92.04.10.03 signed by the responsible of the sponsor and by the experimenters may be found in appendix 1. A pre-study was carried out using one rabbit. On the basis of the results obtained, the study continued using tw o other rabbits. This study took place in accordance with rules of Good Laboratory Practices as * published by : ' ' - The French Ministry of Social Affairs and National Solidarity : State Secretariat Responsible for Health Instructions concerning Good Laboratory Practices (G.L.P.) in the field of experimental toxicology. Instruction of May 31, 1383, Official Text No. 1065 ; Reference SN-S83/25. - Directive 87/18/E.E.C., this directive referring to Recommendation CC81/30 Appendix 2 of the O.E.C.D. - The Food and Drug Administration : G.L.P. 21 C.F.R. Part 58 of December 22, 1978 and amendments of April 11, 1980 and September 4, 1987. Company Sanitized. Does Hot contain tca i CERB - STUDY No. 920084 E 11.4 COMPOUND The technical data and the analytical certificate concerning the compound are attached to the study report in Appendix 2. - Name - Batch number - Record number - Origin - Date received - Amount - Appearance -pH ELF ATOCHEM SA Saturday April 18, 1932 1 bottle of approximately 107 g 11.5 ANIMALS - SPECIES : New Zealand Albino Rabbit. - ORIGIN : CEGAV specialized breeding establishment (Les Hautes Noes, Saint Mars d'Egrenne, 61350 PASSAIS LA CONCEPTION, FRANCE). - WEIGHT : Between 1.5 kg and 2.5 kg at the start of the trial. - SEX : female. - NUMBER : 3. - DATE OF DELIVERY : On 15.04.1992. - ACCLIMATIZATION : For at least 5 days before the trial, in the zone where the experiment was to take place. Animals were selected on the basis of their general condition. Ocular examination took place before treatment. - IDENTIFICATION : Animals were identified individually by ear clipping. - HOUSING : Animals were kept in cages of standard size. Excreta were eliminated by unrolling plastified brown paper which was placed under the cages. These cages were kept in an air-conditioned (17C-21C) animal house, kept at relative humidity between 45 per 100 and 65 per 100 apart from during cleaning, in which non-recycled filtered air was changed approximately ten times per hour. The artificial daylight cycle was 12 hours light and 12 hours darkness. Company Sanitized. Does not contain TSCA CBI CERB - STUDY No. 920084 E 8 - FOOD : (JAR 112 foodstuff. An analytical certificate concerning this foodstuff is included in the study report in Appendix 2. - DRINKING WATER : Drinking water consisted of tap water distributed ad libitum in polycarbonate feeder bottles with a stainless steel teat. A water sample was taken every three months and sent to the Direction des Services Vtrinaires, 216 Rue Louis Mallet, 18014 BOURGES Cedex, FRANCE, for analysis. The water analytical certificate in Appendix 3 corresponds to the sample taken at the closest date of the present study. II.6 ADMINISTRATION OF COMPOUND 0.1 ml of c o m p o u n c / | jj | j| ^ | ^ | ^ ^ J w a s introduced into the conjunctival cul de sac of the left eye of each of the animals. Eyelids were held closed for approximately 10 seconds in order to avoid any loss of the compound. The non-treated right eye served as a control. Following treatment, animals were kept for one hour in a restraint cage in order to prevent them from rubbing the eye, and were then returned to their normal cage. The day of instillation was taken as study day D1. II.7 CLINICAL MONITORING OF ANIMAI S Lesions of the conjunctiva, iris and cornea were monitored by direct examination. Corneal examination was completed by an examination after instillation of an aqueous solution of sodic fluorescein at 2 per 100 and rinse w ith isotonic sterile pyrogen free sodium chloride solution. One hour later then after 24, 48, 72 hours and on D8 of monitoring, conjunctival, iris and corneal lesions were evaluated using the following score system: QjSMpdriy.SantizecL"uses contain TSC.ACB* CERB - STUDY No. 92008 4 E CONJUNCTIVAL LESIONS CHEMOSIS (Swelling) Eyelids and nictitating membrane No swelling Slight swelling, including nictitating membrane Swelling with eversion of eyelid Swelling with eyelids half-closed Swelling with eyelids more than half or completely closed REDDENING OF PALPEBRAL CONJUNCTIVA : Vessels normal Vessels much more injected than normal Vessels difficult to distinguish individually - diffuse bright red color - diffuse dark red color IRIS LESIONS Normal Much more folded than normal, congestion, swelling, iris still reacting to light, even slowly (one or more of these characteristics) No reaction to light, hemorrhage, severe destruction (one or more of these characteristics) CORNEAL LESIONS ABSENCE OF CORNEAL LESIONS No visible modification by direct examination (loss of shine or gloss) or after instillation of fluorescein DEGREE OF OPACIFICATION Presence'of translucent zones (diffuse or disseminated), details of iris clearly visible Presence of easily identifiable translucent zone, details of iris slightly masked Presence of opalescent zone, no detail of iris visible, outline of pupil scarcely discernible Presence of total corneal opacity completely obscuring iris and pupil c\i ro 3 0 1 0 1 2 3 0 1 2 0 1 2 3 4 CERB - STUDY No. 920084 E 10 II.8 CALCULATION OF OCULAR IRRITATION INDICES Mean indices are calculated from results obtained for each rabbit at times 24 hours, 48 hours and 72 hours. The following are noted : - for each parameter, mean of indices scores for each rabbit for the three times (Mean index for each parameter, for each animal for the 3 times = Mi). The theoretical maximum score is 4 (conjunctival edema), 3 (reddening of palpebral conjunctiva), 2 (iris lesions) and 4 (corneal lesions). - for each parameter, the cumulative mean (sum of mean indices for the three animals / 3 = sum of Mi/3). The theoretical maximum score is 4 (conjunctival edema), 3 (reddening of palpebral conjunctiva), 2 (iris lesions) and 4 (corneal lesions). |iW pany Sanitized. Does not contain TSCA CB1 CERB - STUDY No. 920084 E 11.9 STEPS OF THE STUDY 11 A pre-study was carried out using one rabbit. On the basis of the results obtained, the study continued using 2 other rabbits. 11.10 CLASSIFICATION The c o m p o u n c |^ B H I^ ^ ^ ^ ^ r i^ was classified irritant or non irritant on the basis of indices obtained in accordance with the Directive 83/467/E.E.C. proposed by the Official Gazette of the European Community. Compound is found to be irritant if : - cumulative mean (Cm = sum of Mi/3) is higher or equal to the values indicated in table I for at least one parameter. - or if, for at least one parameter, mean indices (Mi) are higher or equal to the values indicated in the table I in at least 2 animals. TABLE I PARAMETER Chemosis Redness Iris Cornea THRESHOLD IRRITATION VALU E 2 2.5 1 2 11.11 DATA ENTRY OF RESULTS All data were recorded as and when obtained using files classified by the study number. .nTSCAC^ it. 'f~ \ - * CERB - STUDY No. 920084 E 12 11.12 REPORT The report of experiment included all findings noted during the course of the study, and in particular : - results of monitoring of animal - ocular irritation indices, - classification of the compound 11.13 QUALITY ASSURANCE The Quality Assurance Unit ensured that working procedures relating to this type of study were strictly adhered to, by periodic inspections of random, over the course of the year. Data included in the report of the experiment were audited by the Quality Assurance Unit, in accordance w ith standard procedures at the Centre. 11.14 RECORDS The protocol, raw data, the correspondance, the report and a sample of the compound have been stored for five years at CERB, 18800 BAUGY, FRANCE, starting from the date of the final report. A t the end of this period, CERB will contact the sponsor in order to jointly determine to either : - continue the records, - return the data to the sponsor, - destroy the data. jpmpany Sanitized. Does not contain TSCA c m CERB - STUDY No. 920084 E 13 III. RESULTS j g j f e o i ManWzg'd, O oja not contain TSCACBi CERB - STUDY No. 920084 E III-1 DEVIATION FROM PROTOCOL N 92.04.10.03 : No deviation from the protocol was recorded during the study. The study was performed according to protocol n 92.04.10.03. III.2 DATES OF TRIAL - Date of start of the pre-study : 04.05.1992. - Date of end of the pre-study : 11.05.1992. - Date of start of the main study : 07.05.1992. - Date of end of the main study : 14.05.1992. III.3 OCULAR TOLERANCF Results of findings are shown in Table II (irritation indices at 24h, 48h and 72h and mean indices per parameter) and III (cumulative means for three animals), as well as in the list of results which may be found in Appendix .5. Evidence fgund approximately one hour then 24 hours after the instillation of c o m p o u n d l H | B | | | ^ ^ | M n o f slight to moderate conjunctival edema and of slight conjunctival redness, in all animals. At times 48 hours and 72 hours, the degree of these lesions tended to decrease, in all animals. No conjunctival lesion reported at time D8. Approximately one hour after the instillation of c o m p o u n c lH i^ ^ ^ m ||^ ^ M th e iris of animal No 920524 was markedly more folded than normal. At time 48 hours, this lesion was present to the same degree in animals Nos 920524 and 920525. No further iris lesion visible at time 72 hours. No lesion of the cornea found by direct examination. At times one hour, then 24 hours, 48 hours or 72 hours, examination after the instillation of fluorescein revealed a fluorescent zone covering between a quarter and half of the corneal surface of the dosed eye in animal Nos 920524 and 920525. At times 24 hours and 48 hours, evidence found of a fluorescent zone covering less than one quarter of the corneal surface of the control eye in these tw o animals. This fluorescence indicated the existence of an erosion of the corneal epithelium which possibly resulted from rubbing by fur or other mechanical trauma (highly sensitive technique). No further irritation reactions seen at time D8. ----- COBipany Sanitized. Does not contain TSCA CBI CERB - STUDY No. 920084 E TABLE II Irritation indices (24h - 48h - 72h evaluations) and mean indices per parameter I0 Reading time 24 H 48 H 72 H MEAN INDEX (Mi) Rabbit C R I C C R I C C R I C C R I C Number H E R 0 H E R 0 H E R 0 H E R 0 ED I RED I RED I R E D I R M S NM S NM S N M SN 920523 1 1 0 0 1 1 0 0 0 1 0 0 0.67 1.00 0.00 0.00 920534 2 1 0 0 1 1 1 0 0 0 0 0 1.00 0.67 0.33 0.00 920525 2 1 0 0 0 1 1 0 0 1 0 0 0.67 1.00 0.33 0.00 CHEM = chemosis RED = redness CORN = cornea TABLEAU III 3 Cumulative means for the three animals - CHEMOSIS REDNESS IRIS CORNEA Sum of mean indices of three animals 2.34 2.67 0.66 0.00 Cumulative mean (Sum/3) = Cm 0.78 0.89 0.22 0.00 LW|>f5r isS fitiizv, Does tint'. :Om<= CERB - STUDY No. 920084 E - Cumulative means (Cm) of mean indices were : . Chemosis : 0.78. . Redness : 0.89. . Iris : 0.22. . Cornea : 0.00. - Mean indices per rabbit (Mi), for the three times taken together were : . Chemosis: 1.00 in one animal and 0.67 in tw o animals. . Redness : 1.00 in tw o animals and 0.67 in one animal. . Iris : 0.33 in tw o animals and 0.00 in one animal. . Cornea : 0.00 in all animals. Classification of the compound is : non-irritant compound for the eye of the Rabbit. Hcontain TSCA CERB - STUDY No. 920084 E 17 IV. CONCLUSION Ocular primary irritation caused in the Rabbit by instillation of compound studied- Under the experimental conditions adopted, the final classification of compound : non-irritant "compound for the eye of the Rabbit. tar T S C A CBS w9n CERB - STUDY No. 920084 E 18 APPENDIX 1 Protocol n 92.04.10.03 Ip B p ffly Sffizea. Does ffot conlaln TSCA CB! CERB - PROTOCOL N 92.04.10.03 Page 1 of 7 TRIAL Ocular primary irritation in the Rabbit SPONSOR: ELF ATOCHEM La Dfense 10 - Cedex 42 92091 PARIS LA DEFENSE - FRANCE SITE OF TRIAL : CENTRE DE RECHERCHES BIOLOGIQUES Chemin de Montifault 18800 BAUGY, FRANCE APPROBATION : SPONSOR CONTACT : Mr REGNIER 22/04/92 Date Signed RESPONSIBLE STAFF FOR CERB : Scientific and Technical Director : S. RICHARD 16/04/92 Date Signed Head of Toxicology Department : C. AUDEVAL-GERARD 16/04/92 Date Signed Study Director : C. BESSON 14/04/92 Date Signed Quality Assurance : C. VIGIER 16/04/92 Date Signed ed. Does hot contain TSCA CB! CERB - PROTOCOL N 92.04.10.03 U Page 2 of 7 II-2 AIM OCULAR PRIMARY IRRITATION Determine th ^ irrjta n tp ro D e rtie s and/or degree of corrosion induced by c o m p o u n d l U ^ ^ ^ ^ m ^ f o ll o w in g single ocular instillation in the eye of the Rabbit. II.3 METHOD Ocular irritation and/or corrosion following instillation of compouncl J ^ ^ i n the conjunctival cul de sac of the left eye of the Rabbit are evaluated by grading of the ocular irritation reactions seen, using a predetermined score system. The protocol used for study of ocular primary irritation, complies w ith requirements of OECD Guideline No. 405 (February 24, 1987) and EEC Directives (67/548, 79/831, 83/467, 84/449) and enables full evaluation of the reversible or irreversible nature of effects seen. This method is not applicable to substances w ith a pH of less than 2 or more than 11.5, nor those found to have a definite corrosive action or to be source of severe irritation during dermal irritation tests, for which a corrosive effect on the ocular mucosa is then predictable. A preliminary study is carried out in one rabbit. If the results reveal no irritation or only slight irritation, the study is continued in tw o other rabbits. If the results reveal an irritant effect (scoring of conjunctival edema or redness > 3 or iris lesions > 2 or degree of opacification > 3) the preliminary study may be extended to D8 and D15 to show possible, reversibility of lesions, but the study will not be repeated in two other rabbits. This study took place in accordance'with rules of Good Laboratory Practices as published by : - The French M inistry of Social Affairs and National Solidarity : State Secretariat Responsible for Health Instructions concerning Good Laboratory Practices (G.L.P.) in the field of experimental toxicology. Instruction of May 31, 1983, Official Text No. 1065 ; Reference SN-S83/25. - Directive 87/18/E.E.C., this directive referring to Recommendation CC81/30 Appendix 2 of the O.E.C.D. - The Food and Drug Administration : G.L.P. 21 C.F.R. Part 58 of December 22, 1978 and amendments of April 11, 1980 and September 4, 1987. not contain TSCA CBI CERB - PROTOCOL N 92.04.10.03 21 Page 3 of 7 I. COMPOUND DATA SUPLIED BY THE SPONSOR : The substance information sheet and the analytical certificate were received on A p rili 0,1992. . DATA TO BE SUPPLIED BY THE SPONSOR : please fill in the test substance information sheet. AMOUNT OF COMPOUND TO BE SypPLIED : 3 ml of c o m p o u n d f p ^ j^ ^ ^ p P jp r e required for the study. IV - GENERAL CHARACTERISTICS OF STUDIES : IV-1 ANIMALS : -SPECIES: New Zealand Albino Rabbit. , -ORIGIN : From CEGAV specialized breeding establishment (Les Hautes Noes, Saint Mars d'Egrenne, 61350 PASSAIS LA CONCEPTION, FRANCE). -W E IG H T: Generally between 1.5 kg and 2.5 kg at the start of the trial. - SEX : male or female. -NUMBER : 3. -ACCLIMATIZATION: ' ' For at least 5 days before the trial, in the zone where the experiment is to take place. Animals are selected on the basis of their general condition. Ocular examination takes place before treatment. ' -IDENTIFICATION : Animals are identified individually by ear clipping. -HOUSING: , Animals are kept in cages of standard size. Excreta are eliminated by unrolling plastified brown paper which is placed under the cages. p S p a iy SanlilzetJ. B oesnol contain TSCA CBI CERB - PROTOCOL N 92.04.10.03 00 Cm L Page 4 of 7 These cages are kept in an air-conditioned (17C-21C) animal house, kept at relative humidity between 45 per 100 and 65 per 100, apart from during cleaning, in which non-recycled filtered air is changed approximately ten times per hour. The artificial daylight cycle is 12 hours light and 12 hours darkness. - FOOD : UAR 112 foodstuff. An analytical certificate concerning foodstuff is included in the study report. - DRINKING WATER : Tap water distributed ad libitum in polycarbonate feeder bottles with a stainless steel teat. A water sample is obtained every three months and sent to the Direction des Services Vtrinaires, 216 Rue Louis Mallet, 18014 BOURGES Cedex, FRANCE for analysis. A water analytical certificate is included in the study report. IV-2 ADMINISTRATION OF COMPOUND 0.1 ml of c o m p o u n d n ^ m m i m i i s introduced into the conjunctival cul de sac of the left e y e ^ o ^ e a c f ^ o ^ t f ^ animals. Eyelids are held closed for approximately 10 seconds in order to avoid any loss of the compound. The non-treated right eye serves as a control. Following treatment, animals are kept for one hour in a restraint cage in order to prevent them from rubbing the eye, and are then returned to their normal cage. The day of instillation is taken as study day D1. Study stages : - A preliminary study is carried out in one rabbit. . If the results reveal no irritation or only slight irritation, the study is continued in tw o other rabbits. . If the results reveal an irritant effect (scoring of conjunctival edema or redness > 3 or iris lesions > 2 or degree of opacification > 3) the preliminary study may be extended to D8 and D15 to show possible reversibility of lesions, but the study will not be repeated in tw o other rabbits. IV-3 CLINICAL MONITORING OF ANIMALS Lesions of the conjunctiva, iris and cornea are monitored by direct examination, and can be continued using an examination lamp. ' Corneal examination will be completed by an examination after instillation of an aqueous solution of sodic fluorescein at 2 per 100 and rinse w ith isotonic sterile pyrogen free sodium chloride solution. jP s j & i f t f ' s o ' m a l r t TsCA C CERB - PROTOCOL N 92.04.10.03 Page 5 of 7 One hour later then after 24, 48, 72 hours (and on D8 of monitoring and D15 if irritation reaction are reported at 72 hours) conjunctival, iris and corneal lesions are evaluated using the following score system : CONJUNCTIVAL LESIONS CHEMOSIS (Swelling) Eyelids and nictitating membrane No swelling Slight swelling, including nictitating membrane Swelling w ith eversion of eyelid Swelling w ith eyelids half-closed Swelling w ith eyelids more than half or completely closed 0 1 2 3 4 REDDENING OF PALPEBRAL CONJUNCTIVA : Vessels normal Vessels much more injected than normal Vessels difficult to distinguish individually - diffuse bright red color - diffuse dark red color 0 1 .2 3 IRIS LESIONS Normal Much more folded than normal, congestion, swelling, iris still reacting to light, even slowly (one or more of these characteristics) No reaction to light, hemorrhage, severe destruction (one or more of these characteristics) 0 1 2 CORNEAL LESIONS ABSENCE OF CORNEAL LESIONS No visible modification by direct examination (loss of shine or gloss) or after instillation of fluorescein 0 DEGREE OF OPACIFICATION ' Presence of translucent zones (diffuse or disseminated), details of iris clearly visible Presence of easily identifiable translucent zone, details of iris slightly masked Presence of opalescent zone, no detail of iris visible, outline of pupil scarcely discernible Presence of total corneal opacity completely obscuring iris and pupil -! IV-4 CALCUI ATION OF OCULAR IRRITATION INDICES ' 1 2 3 4 ' Mean indices are calculated from results obtained for each rabbit at times 24 hours, 48 hours and 72 hours. - Company Sanitized. Does not contain TSCA CERB - PROTOCOL N 92.04.10.03 2 Page 6 of 7 The following are noted : - for each parameter, mean of indices scores for each rabbit for the three times (Mean index for each parameter, for each animal for the 3 times = Mi). The theoretical maximum score is 4 (conjunctival edema), 3 (reddening of palpebral conjunctiva), 2 (iris lesions) and 4 (corneal lesions). - for each parameter, the cumulative mean (sum of mean indices for the three animals 13 = sum of Mi/3). The theoretical maximum score is 4 (conjunctival edema), 3 (reddening of palpebral conjunctiva), 2 (iris lesions) and 4 (corneal lesions). IV-5 CLASSIFICATION : The c o m p o u n d U | | | i m H M < s classified irritant or non irritant on the basis of indices obtained in accordance with the Directive 83/467/E.E.C. proposed by the Official Gazette of the European Community. Compound is found to be irritant if : - cumulative mean (Cm = sum of Mi/3) is higher or equal to the values indicated in table I for at least one parameter. - or if, for at least one parameter, mean indices (Mi) are higher or equal to the values indicated in the table I in at least 2 animals. TABLE I PARAMETER Chemosis Redness Iris Cornea THRESHOLD IRRITATION VALUE 2 2.5 1 2 V - RESULTS: All data are recorded as and when obtained using files classified by the study number. 11.12 REPORT : The report of experiment includes all findings noted during the course of the study, and in particular : - resu..l.t.s...o..f...m...o..n..i.t.o...r.i.n9g of animals, - ocular irritation indices, a*---. c o e - s n o t aon.ism, iSUAUfeii *>?V..a.!-***'-0 " - classification of the study compound. CERB - PROTOCOL N 92.04.10.03 25 Page 7 of 7 VII - QUALITY ASSURANCE The Quality Assurance Unit ensures that working procedures relating to this type of study are strictly adhered to, by periodic inspections of random, over the course of the year. Data included in the report of the experiment are audited by the Quality Assurance Unit, in accordance with standard procedures at the Centre. VIII - RECORDS The protocol, any possible amendments, raw data, the correspondence and the report, and a sample of the compound are stored for five years at CERB, 18800 BAUGY, FRANCE, starting from the date of the final report. A t the end of this period, CERB will contact the sponsor in order to jointly determine to either : - continue the storage of the records, - return the data to the sponsor, - destroy the data. IX - TIMETABLE : Date of start of study : Week of May 04, 1992. Results by telex : at the end of the study. Date of submission of the draft report : first fortnight of June 1992. Date of submission of the English summary : at the final report submission. Timetable assured if compound reaches CERB 15 April 1992 at the latest. Company Sanitized. Does not contain TSCA CBI CERB - STUDY No. 920084 E 26 APPENDIX 2 Technical data and analytical certificate concerning compound 'party Jam'Hzed. Does rtc! cc itair rscA1 SERVICE DE TOXICOLOGIE CONFIDENTIEL Avril 1992 elf atochern sa La Dfense 10, cdex 42 92091 Paris-La Dfense France FICHE D'INFORMATIONS IDENTIFICATION Nom commun Nom chimique Origine; n de lot N d 8archivage (CAL) PROPRIETES PHYSICO-CHIMIQUES Apparence Viscosit Temprature bullition pH Densit Point d'clair Solubilit 3 INFORMATIONS TOXICOLOGIQUES ET PRECAUTION D'EMPLOI Aucune information disponible._____________________ CONDITIONS DE CONSERVATION ET DE DESTRUCTION Conservation Stabilit Destruction : Al'obscurit et 1'abri de la chaleur. : Stable jusqu'en Avril 1993 dans ces conditions de stockage : Incinration. Company Sanitized. Does not contain TSC CBI CO elf atochem sa 4, cours Michelet, cedex 42 92091 Paris La Dfense 10 France BULLETIN D 'ANALYSE 2 AT Jean-Franois Rgnier Service de Toxicologie # J S M f a S c - j $ n o co n l ri 3CA cm CERB - STUDY No. 920084 E 29 APPENDIX 3 Analytical certificate concerning foodstuff Igompany Sanitized. Does net contain TSCA CB1 30 /""N y ALIMENT COMPLET LAPINS - ENTRETIEN Granuls : 4,5 mrr Ration journalire du Lapin: suivant la race, de 100 150 g -- Eau volont. FORMULE % Crales, s u c re ..................................... 42,8 Issues et lgumineuses ........................ 49 Protines vgtales (to u rte a u x).......... 4,2 Compos minral vitaminis .............. 4 ANALYSE MOYENNE % Valeur calorifique (C a l/k g ).............. . 2.200 Eau' ......................................... ......... 10 Protides ............................................. - 13 .Lipides .............................. ................. Glucides (E.N.A.7 "v,............................ 2.7 49,3 Cellulose (Weende) ' ....................... 17' Minraux .................................. : 8 ACIDES AMINES . (calculs en m g/kg) Arginine ........' . l ' . l ...................... . 6.800 C s tin e ............................................ 2.100 Lysine ..................... 4.600 Mthionine ........................... 1.600 Tryptophane ......... 1.400 G ly c in e ............ , ................. , . '___ _ 5.200 j MINERAUX i (calculs en* mg/kg) i ` App. Nat. (moy.) App. par CM TOTAUX ; i p ................................. | 0 3 ............................ !1 K ........... ..................... : N a ....................,...... 4.100 4.500 10.600 .1.150 3.500 4.600 0 1.950 . Mg ........................... 2.100 130 Mn .......................... 40 40 ! F j Cu I Zn ............. .............. ............................ ............................ 160 12 30 ! 140 IS 45 ' C o ........... o, ; 1.5 7.600 9.100 10.600 3.100 . 2230 BO 300 27 7s .6 ; : : .! ` : ; : ; i \ 1 ................................. apport 6 0 us 'orme assimilable par algues marines. VITAMINES (calcules au kg) App. Nat (moy.) App. Syntht TOTAUX Vitam. A .... 2 8 5 0 Ul 0 3 . 30 . - B1 . 4.30 - B2 . 3.80 . . B 3 . 16 . . B6 . 1- E ... 16 - K3 . 6- . P P ... C h o l i n e .... 55 850 - 6.500 Ul 9 3 5 0 Ul 1.000 - 1.0 3 0 . . 0,00 m g 4,30 m g 0.00 - 3.80 0.00 . 1. 16 2. 10 - 26 1- 7* s- 60 - 200 - 1.050 - Company Sanitized. Does not contain T fijBS cn CERB - STUDY No. 920084 E APPENDIX 4 - : . Analytical certificate concerning water CCNCKAlOU LABORATOIRE DPARTEMENTAL V T R IN A IR E 32 RESULTAT D'ANALYSE D'EAU Prlvement d 'eau parvenu le: Lundi 17 Fvrier i<m E ffe c tu p a r: E .R .B . O rig in e : Robinetdnoooloirpmpre. A g e n t le client UNTTF, T,AP1N COR AYFS Commune: BAUGY Physicochim ie Rsultats Nonnes Bactriologie Tutbidit en U Jackson Conductivit pS/cm PH Qxydabilit KMnCW mg/l Duret en "Franais TA.C en "Franais Ammoniaque en mg/I Nitrites en mg/i Nitrates en mg/l Sulfates en mg/l Chlorures, en mg/I Fer en mg/i Manganse en mg/I Fluor en mg/I - : : : : : : : : : : 0 .4 639 7 .5 0 .8 34 24 0 0 S9 64 29 0 <2U 6 .5 -9 <5 < 0.5 < 0.1 < 50 < 250 < 200 < 0 .2 < 0.05 < 1.5 Conformes thermotolrants/fOOml Streptocoques fcaux/1O0m( Coliformes/100ml Dnombrement des bactries arobies revivtfiables 37C/ml ' ' 22"C/ml Spores de bactries anarobies suffrto-rductrices/20m! ' * ` /100ml Salm o nelles Staphylocoques pathognes <5 *:S e lo n < le c re t N ^8$r3.et su ivants. Rsultats O 0 0 <1 <1 Nonnes* 0 0 0-95% if 0 0 0 0 B ourges.Ie: Lundi 24 Fvrier 1992 L e Director duLaboratoire Je a n M arie GUERATTD Pompany Santfizetf. Does not contain TSCA CF CERB - STUDY No. 920084 E 3 APPENDIX 5 Results data sheets ernpany SsnYizad. Ooes noi confai YSCA Cf CERB . 3 U0 0 IN D IC E D r IR R IT A T IO N O C U LAIR E PRODUIT CHIM IQ UE UJ, ESPECE : L a p in No - Z la n d a is SEXE : a i e - / fe m e lle VOLUME o u MASSE A D M IN IS T R E E : DEBUT D 'ETUDE F IN D ' ETUDE : L}.O S ^ : ' f j q, r -<^ 'b HEURE DE DEBUT DE L 'IN S T IL L A T IO N / PARAPHE : i f S / cj> 0S HOURS NAEEO IAEIN 24 H 48 H EKE -0$ 3>L EKIE : C O ^ l CR Ic HOR 0 EU IR MG S N CRI HOR Eu I MGS c 0 R N 72 H EKE : r-C ^ b t CR I C H0R 0 Eu I R MGS N &rucGV> A1 0 Q i i 0 o G 1 TDQffil/nCOECE IN2EOE . /SNIMKL . MD5CEN RI C OR 0 UI R GSN Ml00 OO 0.00 S G -C 1 ESEfflE BEDES W z o / c f y to o /& ci es. CBSEI5EDCNS DIVERSES ' itqL JLC A ^ 0n' cU - ccfuMJii^ -fckiC -COJhc w iMtan. o itC IHMES POURS NUEEO IEIN ' 1H /./a S A l R O U G 1?>u>&3 1 Oo J8 -ii./o S /^ P J 15 C R Ic CRIC H 0 R0 H 0 R0 E U I R E U-I R M G SN M G SN 0 <7 EREAEHE HEURE % ( 1 1 't / c 5 p-foo / cb / Company Sanitized. Does not contain TSCA CBI ERB N ETUDE : )bO O PRODUIT : N LOT IN D IC E D ' IR R IT A T IO N O CULAIRE PRODUIT CHIM IQ UE ESPECE : L a p in N o - Z la n d a is SEXE : ja ie - / fe m e lle DEBUT D ' ETUDE VOLUME e u MASSE A D M IN IS T R E R : O ) F IN D 'E TU D E zjLoS^Z. HEURE DE DEBUT DE L 'IN S T IL L A T IO N / PARAPHE : UfcTES LECHEE NMD IfiPIN yu o ii jo o S tS 24 H LATE : Zo ^ l CR I C H0 R 0 EU I R MG SN 2) / 0 O b/ 0 O 48 H EME j 9o*>% CR H0 EU MG i R I S c 0 R N 72 H DKIE : /-oS^Z CR I C H0 R 0 EU I R MGSN A / I A U-- G O > O 0 /I A 0 OO O TOEtM/QLirCE RDTOE' /NIraL A MM c R i cc R I C H 0 R 0H 0 R 0 E U 1 RE U I R M G S NM G S N Jrv 6% 461- 0 /ca oco 441 2 2 A 0 l,00 Oli Cc cube?! EHE&Effi /diH<3o HEURE / jV> AM I y / Yv A If ho 2> / Ci ci T0ES x la IPCDJFE t. /ab/Z>Z NUMERO IfiPIN c R Ic H 0 RO E U IR M G SN J8 A u . M . /fz_ J1S c R i c CR IC Q 0 R 0 H 0 R0 E. U - I R E U I R M G SN M G SN VAi0 0 0 l 1 0 0. J ^O . -Z) A . BPRKEHE HEURE m 'r > / c h A 1% / A / 1 1 ~<5->t.' ary SanitiIK. io ! fiOlltcin i'Oa