Document g2kYJoXeXZdLpX6DoGNM5514a
%^J W
AR226-2993
Baugy, June 17, 1993
STUDY No 920084 E
OCULAR PRIMARY IRRITATION
Sponsor: ELF ATOCHEM SA La Defense 10 Cedex 42 92091 PARIS LA DEFENSE FRANCE
CompanySanitized. Does not contain T.;
CERB - STUDY No. 920084 E TRIAL COMPOUND
: Ocular primary irritation.
SPONSOR SPONSOR CONTACT ADDRESS
SITE OF TRIAL
ELF ATOCHEM SA
MrREGNIER
La Defense 10 Cedex 42 92091 PARIS LA DEFENSE - FRANCE
CENTRE DE RECHERCHES BIOLOGIQUES Chemin de Montifault
18800BAUGY, FRANCE
RESPONSIBLE STAFF FOR CERB
Scientific and Technical Director :
S. RICHARD Pharmacien Docteur de Seme Cycle MaTtre en Pharmacologie
oniA^-i-a ^
Date
Signature
Head of Toxicology Department :
C. AUDEVAL-GERARD Docteur Veterinaire CES d'Ophtalmologie
5o^oG/55
Date
Signature
Study Director : C.-BESSON D.U.T. de Biologic Appliquee
Quality Assurance : C. VIGIER
Date Date '
Signature
^^~f^ \ j Signature
The present report is based upon the actual and authentic experimental results
obtained.
"o^oim'S'anlU;^, 'Ooe;; Sot fioMa, TSCA CBi
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 11.2 11.3 11.4 11.5 11.6 11.7 11.8 11.9 11.10 11.11 11.12
11.13 11.14
INTRODUCTION AIM METHOD COMPOUND ANIMALS ADMINISTRATION OF COMPOUND CLINICAL MONITORING OF ANIMALS CALCULATION OF OCULAR IRRITATION STEPS OF THE STUDY CLASSIFICATION DATA ENTRY OF RESULTS REPORT QUALITY ASSURANCE RECORDS
INDICES
III.
RESULTS
111.1 DEVIATIONFROMPROTOCOLNo92.04.10.03
111.2 DATES OF TRIAL 111.3 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
companySanitized. Does not contain TSCA CBI
CERB - STUDY No. 920084 E
I. SUMMARY
KI ^
The aim of this study was objective determination of the irritant properties and/or
compoundfm^Bm degree of corrosion induced by
!innts'tt-;illl"lsa+t;io->ln-i iirn>+t<o"> the +1->< eay^e> oi^ff 4t-h^>en DRaikbkb:i+t.
1^
{following single
^
0.1 ml of compounclimB|^^|H^vere instilled into the conjunctiva) cul de
sac of the left eye of^he 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 two animals.
. Redness : 1.00 in two animals and 0.67 in one animal.
. Iris
: 0.33 in two animals and 0.00 in one animal.
. Cornea : 0.00 in all animals.
/as found to be non-irritant for the eye of the Rabbit. ^i?!pSnya<irSi.?.2d.DDSSftot contain 7SCA CB?
CERB - STUDY No. 920084 E
II. EXPERIMENTAL PROTOCOL ADOPTED
Ife
CERB - STUDY No. 920084 E
'0
11.1 INTRODUCTION
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, 79/83.1, 83/467, 84/449) and enabled full evaluation of the
reversible or irreversible nature of effects seen.
11.2 AIM
The aim of the study was objective determination of the irritant properties and/or
degree of corrosion induced by compoundHH||B|^mH||HBB|^
11.3 METHOD
1flf"in Ouccuuilar irritation and/or corrosion following instillation of compoundl the conjunctival cul de sac of the left eye of the Rabbit were evaluated by gradinng of the ocular irritation reactions seen, using a predetermined score system.
This method was not applicable to substances with 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 rabbits.
study
continued
using
two
other
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, 1983, Official Text Mo. 1065 ; Reference SN-S83/25. - Directive 87/18/E.E.G., 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.
companySanTtized. Does ftol contain T<SC A *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
11.5 ANIMALS
: ELF ATOCHEM SA : Saturday April 18, 1992 : 1 bottle of approximately 107 g
- 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.
'gCcrmpaSnaynitized. Does not contain TSCA CBI
CERB - STUDY No. 920084 E
- FOOD : UAR 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
Veterinaires, 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.
- 11.6 ADMINISTRATION OF COMPOUND
0.1 ml of compounqlU^UUljwas introduced into the conjunctiva! cul
de sac of the left e^e 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 ANIMALS
Lesions of the conjunctiva, iris and cornea were monitored by direct examination.
Cornea! examination was completed by an examination after instillation of an aqueous solution of sodic fluorescein at 2 per 100 and rinse with isotonic sterile pyrogen free sodium chloride solution.
One hour later then after 24, 48, 72 hours and on D8 of monitoring, conjunctiva!, iris and corneal lesions were evaluated using the following score
system:
^jmany ^awizeci- uses not conissn TSCA KB<
CERB - STUDY No. 920084 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
0
Much more folded than normal, congestion, swelling, iris still
reacting to light, even slowly (one or more of these characteristics)
1
No reaction to light, hemorrhage, severe destruction (one or more
of these characteristics)
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
1
Presence of easily identifiable translucent zone, details of iris
slightly masked
2
Presence of opalescent zone, no detail of iris visible, outline of pupil
scarcely discernible
3
Presence of total corneal opacity completely obscuring iris and pupil
4
3\
r
CERB - STUDY No. 920084 E
1 Q
11.8 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 (conjunctiva! 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 (conjunctiva! edema), 3 (reddening of palpebral conjunctiva), 2 (iris lesions) and 4 (corneal lesions).
^^^e-.n.-O.^TSCACB,
CERB - STUDY No. 920084 E
11.9 STEPS OF THE STUDY
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 compouncj^^H^^^^BBjwasclassified 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
11.11 DATA ENTRY OF RESULTS
All data were recorded as and when obtained using files classified by the study number.
^ar.-^^-i^.' s^0""*''6"^ -
CERB - STUDY No. 920084 E
^Hy
11.12 REPORT
The report of experiment included all findings noted during the course of the study, and in particular:
- results of monitoring of animals, - ocular irritation indices, - classification of the compoun
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 with 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.
At 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.
tempany Sanitized. Does not contain TSCA CRI
CERB - STUDY No. 920084 E
, ..
III. RESULTS Sanito'd, goes not cpntein TSCACBl
CERB - STUDY No. 920084 E
1 4 111.1 DEVIATION FROM PROTOCOLN 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.
II.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 TOLERANCE
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 found approximately one hour then 24 hours after the instillation of
compoundf||UB|fHBi)of 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 conjunctiva! lesion reported at time D8.
Approximately one hour after the instillation of compounqfH
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 instillation of fluorescein and half of the corneal 920525.
24 hours, 48 hours or 72 hours, examination after the
revealed a fluorescent zone covering between a quarter
surface of the dosed eye in animal Nos 920524 and
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 two
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.
,,C_o,,m.-p.a. ny.S.,ani.t.ized, .
,,
Does
not.,
, . contain
_TS-C.,A._CBI
15 CERB - STUDY No. 920084 E
TABLE II
Irritation indices
(24h - 48h - 72h evaluations) and mean indices per parameter
Reading time
24H
48H
72H
MEAN INDEX (Mi)
Rabbit
C R
C C R
CC R
C C
R
I
C
Number H E R 0 H E R 0 H E R 0 H
E
R
0
ED I R ED I RED I R
E
D
I
R
M
S NM
S NM
SN M
S
N
920523 920534 920525
1 1 0 0 1 1 0 0 0 1 0 0 0.67 1.00 0.00 0.00 2 1 0 0 1 1 1 0 0 0 0 0 1.00 0.67 0.33 0.00 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
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
'%W{5ny ^aft'ifa"^ 'Does nn^'ht^i T^C^
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 two animals.
. Redness : 1.00 in two animals and 0.67 in one animal.
. Iris
: 0.33 in two 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.
^^^n^c.
CERB - STUDY No. 920084 E
IV. CONCLUSION
Ocular primary [rritation caused in the Rabbit by instillation of compound /as studied.
Under the experimental conditions adopted, the final classification of compound
'compound for the eye of the Rabbit.
'
^^S_M_^^a_!w."..^'--m,-^'^^;^:^'"c9pntam TSCA CBS
CERB - STUDY No. 920084 E
APPENDIX 1
Protocol n 92.04.10.03 JRSfflpeSHanyitized. Does not contain TSCA CBI
CERB - PROTOCOL N 92.04.10.03
TRIAL
Ocular primary irritation in the Rabbit
SPONSOR:
ELF ATOCHEM La Defense 10 - Cedex 42
92091 PARIS LA DEFENSE -
FRANCE
SITE OF TRIAL
: CENTRE DE RECHERCHES BIOLOGIQUES
Chemin de Montifault
18800 BAUGY, FRANCE
APPROBATION :
Page 1 of 7
SPONSOR CONTACT :
Mr REGNIER
22/04/92
RESPONSIBLE STAFF FOR CERB
Date
Signed
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
1 6/04/92
Date
Signed
Bi^^anIHzed. Does nol contain TSCA CBI
CERB - PROTOCOL N 92.04.10.03
2.-'.
u Page 2 of 7
OCCUULLAAFR PRIMARY IRRITATION
I12AIM
Determine the irritant properties and/or degree of corrosion induced by
compoundjflflHiH----following single ocular instillation in the eye of the
w
*
Rabbit.
II.3 METHOD
Ocular irritation and/or corrosion following instillation of compounqlHBIHBr
fHJin the conjunctiva! 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 with 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 with 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 two other rabbits. If the results reveal an irritant effect (scoring of conjunctiva! 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 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, 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.
, Sanltfce-d. Does not contain TSCA CBI
CERB - PROTOCOL N 92.04.10.03
21
Page 3 of 7
DATA SUPLIED BY THE SPONSOR : The substance information sheet and the analytical certificate were received on April 10,1992.
DATA TO BE SUPPLIED BY THE SPONSOR : please fill in the test substance information sheet.
AMOUNT OF COMPOUND TO BE SUPPLIED :
3 ml of compoundjUB------^IVr9 ^l11'1'6^ tor 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).
-WEIGHT: 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
place. Animals are selected on the basis of examination takes place before treatment.
where the experiment is their general condition.
to take Ocular
-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. 'EgfflpangSanfflzea. Does nol contain TSCA CBI
CERB - PROTOCOL N 92.04.10.03
22
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 Veterinaires, 216 Rue Louis Mallet, 18014 BOURGES Cedex, FRANCE for analysis. A water analytical certificate is included in the study report.
IV-2 ADMINISTRATION OF COMPOUN
j^BIHI^^HIl ^i
0.1 ml of compound
is introduced into the conjunctiva! cul de
sac of the left eye of each of the 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 two 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 D1? to show possible reversibility of lesions, but the study will not be repeated in two 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 with isotonic sterile pyrogen free sodium chloride solution.
^^^n^rttSeg'^^^
"r^CAt^
CERB - PROTOCOL N 92.04.10.03
23
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 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
0 1 2 3 4
0 1
2 .,,
3
IRIS LESIONS
Normal
0
Much more folded than normal, congestion, swelling, iris still
reacting to light, even slowly (one or more of these characteristics)
1
No reaction to light, hemorrhage, severe destruction (one or more
of these characteristics)
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
1
Presence of easily identifiable translucent zone, details of iris
slightly masked
2
Presence of opalescent zone, no detail of iris visible, outline of pupil
scarcely discernible
3
Presence of total corneal opacity completely obscuring iris and pupil
4
/
IV-4 CALCULATION OF OCULAR IRRITATION INDICES
l) Mean indices are calculated from results obtained for each rabbit at times
24 hours, 48 hours and 72 hours.
- fcompanySanlllze'd. Does not contain TSCA CB1
2
CERB - PROTOCOL N 92.04.10.03
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 / 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).
IV-5 CLASSIFICATION :
compoundlfmUlis The
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
indicated in table I for at least one parameter. - or if, for at least one parameter, mean indices (Mi) are higher
values indicated in the table I in at least 2 animals.
to the or equal
values to the
TABLE|
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 ail study, and in particular : - results of monitoring of animals, - ocular irritation indices, - classification of the study compound.
findings noted during the course of the
^iSll&S8^^^ro^notaomamTSCACBI
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.
At 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 :
w
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
APPENDIX 2 Technical data and analytical certificate
concerning compound
|;oriipany'Safti'iJi?.ea".Does not con'.a'in TSCA C81
SERVICE DE TOXICOLOGIE
CONFIDENTIEL
Avril 1992
27
elf atochehi sa
La Defense 10, cedex 42 92091 Paris-La Defense France
FICHE D'INFORMATIONS
IDENTIFICATION
Norn conunun
Nom chimique
Origine; n0 de lot
N d'archivage (CAL)
INFORMATIONS TOXICOLOGIQUES ET PRECAUTION D'EMPLOl
Auczine informa-fcipn disponible.
____________
CONDITIONS DE CONSERVATION ET DE DESTRUCTION
Conservation
Stabilite
Destruction
: A 1'obscurite et a 1'abri de la
chaleur.
: Stable jusqu'en Avril 1993 dans ces
conditions de stockage
: Incineration.
company Sanitized. Does not contain TSCA CB1 \y1
elf atochem sa
4, cours Michelet, cedex 42 92091 Paris La Defense 10
France
BULLETIN D*ANALYSE
I
..Q^-Kvril 1992
>'
Jean-Francois Regnier Service de Toxicologie
e
1;oTrtr'anv?'Saf;^^.^. 'Oc.'ia HQt coi..if TSCA CT?
CERB - STUDY No. 920084 E
29
APPENDIX 3 Analytical certificate concerning foodstuff
IgompanSyanitized. Does not contain TSCA CB1
30
ALIMENT CQMPLET
LAPINS - ENTRETIEN
Granules: 4,5 mn-
Ration journaliere du Lapin: suivant la race. de 100 a 150 g -- Eau a volonte.
FORMULE %
.............. Cereales. sucre
42,8
....... Issues et .legumineuses
49
Proteines vegetales (tourteaux)
4.2
Compose mineral vitaminise ..
4
ANALYSE MOYENNE %
Valour calorifique (Cal/kg) ........ Eau ............................ Protides .........................
.Lipides .........................
Glucides (E.NAT "',...............
Cellulose (Weende) '....-.......'.. Mineraux ...................'...
. 2.200 10
-13
2.7 49,3 17"
8
ACIDES AMINES
(calcutes en nig/kg)
Arginine ... Cystine .... Lysine ....
Methionine
Tryptophane GlySine
....
6.800 2,100 4.600 1.600 1.400 5.200
j
MINERAUX
i
(calcules en- mg/kg)
i
Aw. Nat.
(<"oy.)
App. par CM
TOTAUX
pr -
Ca .............. K................ Na .............. Mg .............. Mn .............. Fe .............. Cu ..............
4.100 4.500 10.600 .1.150 2.00
40 160
12 30
3.500 4.600
0
1.950 130 40 140 15 45
7.600
9.100 10.600
3.100 . .
2.230 BO
300
27
75
:
Co ..............
0.1
1.5
1.6
:
apporti sous fonne assimilable par alguea
.
V1TAMINES
(calcultes au kg)
:
App. Nat (moy.) App. Sylrthtl
.;.;;.
TOTAUX
Vitam. A .... 2.850 Ul
. D"3 .
30
B 1 .
2' B . . B3 .
4.30 3,80 . 16
. B6 . 1
... E
16
K 3
6
.
. PP ... 55
Choline ..... 850
6.500 Ul 1.000
0,00 mg 0.00 . 0.00
1 10
1 5
200
93SO Ul 1.030.
4.30 rng 3.80 16 2 26 7 60 1.050
D
pompanySanitized. Does not contain Ts6A CPU
w^wy^^'^1
E''--ipX^)C'-^ TIW''rc ^V-<-i'-^"WtWi^g <*V?^A:^^ '- ^f t-t'iy^rTif ole --f'S"' ^'-*) ?u- ? '
CERB - STUDY No. 920084 E
31
APPENDIX 4-.-
Analytical certificate concerning water
|[?giapan^:SanitK&D<oes not contain TSCA GBI
GCNCKAt. OU
LABORATOtRE OEPARTEMENTAL VETERINAIRE
32
RESULTAT D'ANALYSE D^EMT
agr66 par (e Minlstfera de la sant6
Nd'AnaIyse: 920282 -Typed'analysK B3+np
N0 d'Ordre: 394
Prelevement d'eau parvenu Ie: Effectupai: CE.R.B-
O/Mr^igS^inoe. :- rR>ob-inr-t.d a.pttp- l-oir-proprp
Usol aNTTF.TAPINCOBAYF?;
Commune: BAT3GY
Lmidil7Kvrierl992 Agent:
PIiysicochimie
Tu(bidit6 en U Jackson Conducttvft^ pSfcni PH
Oxydabilit^ KMn04 mg/1
Dwet6 en "Franyais
T-A.C en 'Franyais Ammoniaque en mg/I
Nitrites en mg/1 Nitrates en mg/1
Sutfates en mg/1
Chlorures.en mg/I
Fer en ing/I
Manganese en mg/I
'-.
Ftuor en mg/1
Restdtats Normcs* Bacteriologie
: 0,4 639
<2U
Colfformes thermotolerants/IOOmI :
Streptocoques fecaux/IOOmI
:
: 7.5
6.5-9 Colitonnes/IOOmI
:
: 0,8
:
34
<5 Denombremant des bacteries
aerobies revivtftabfes a 37C/ml
:
24
:
0
:
0
:
S 9
<0.5
<0.1 <50
a22C/ml :
Spores de bacteries anaerobies
suffrto-reductrices/20ml "
:
.
/100ml
:
: 64
<250 Sahnonelles
:
29
<200 Staphytocoques pathogenes
:
:
0
<0^
< 0.05
:
< 1.5
Resultats
0 0 o
-
<1
0
Nonnes*
0 0
0-95%
... . -.i - -
$1
0 0 0 0
<5
#-C'n1~-- ^-------it.i0fm
Bourges.le: Luadi 24 Fevrier 1952 Lc Dircctsar du Laboratoiic
J^jS.--^pT---^^---jL^..^
Jean Marie GUERAUD
iCompanySant'ffzed. Does not contain TSCA CP
CERB - STUDY No. 920084 E
3 .
J i-
APPENDIX 5 Results data sheets
:.-'.:?Tipany' 3an;'?izad. L'-;;;i noi con?ai:-,'! SCA Cf
CERB
INDICE D'IRRITATION OCULAIRE
PRODUIT CHIMIQUE
N ETUDE : ^^00 ^
PRODUIT :
ESPECE : Lapin Neo - Zeiandais SEXE : -softie-/ femelle
N LOT
:
DEBUT D' ETUDE : Lf.O^-0^
VOLUME OU
MASSE ADMINISTREE:
FIN D^ETUDE
: ^ Q ^.'^ 7^/
HEURE DE DEBUT DE L' INSTILLATION / PARAPHE : 'Hf/C'->/ <-j>
TEres EECUBE
M1GED IfiEON
W^
24H
IKIE i^-oS^
C
R
I
c
H 0
R
0
E
U
I
R
M G
S
N
^^G 0
48H DME : G.o'^-yL
c
R
I
c
H
0
R
0
E
U
I
R
M
G
S
N
-r { 0 0 -
72H
mns: ^-o95t
c
R
I
C
H
0
R
0
E
U
I
R
M
G
S
N
i />
^
(5 0
TDOffiL/ItOKE IN2ECE
:
.
/PNSVL
4;
M3SEN
I- C R
cC R I C
H 0 R 0H 0 R 0
E U I RE U I K
M G S NM G S N
Z 3 0
0 ^ /,oo 0,00 Q.t0 Sfl
EeSE3E
wrsf.
WiO/C.^>
^HiV^
^A6
c4
cs.
CBEEESSSBSGDNEBSS
fey^.^.W^"^ o^ ^<^^
-^ia6 -ca//)c Aff ]U(M. c1^1 <"c(-('
TEMPS TH.;HH:;
MMESD LBEIN
. 1H
^ /a5A~i
c
R
I c
H
0
R 0
E
U
I R
M
G
S N
9^3 ^ ^ 0 0
J8
-^. y^/^
c
R
I c
H
0
R 0
E
U
I R
M
G
S N
J35
../../
CR IC
H
0
R 0
E
U- I
R
M G SN
0 0 0 (7
EMaHE
HaLEE iwrt-/c^ Woo / ob
/
company Sanitized. Does not contain TSCA CB1
CERB
,,. ^o, ._
PRODUIT ;
%WS^
N0 LOT
;
INDICE D'IRRITATION OCULAIRE
PRODUIT CHIMIQUE
ESPECE : Lapin Neo - Zeiandais
SEXE : maie-/ femelle
DEBUT D''ETUDE
: -}-.0~> ^ 2
VOLUME eu KAS5E ADMINISTRE^: U/ HEURE DE DEBUT DE L'INSTILLATION /
FIN D'ETUDE
PARAPHE : ^HCO/C-'3
:^.^S-"*7'L
1BSS
LECILBE
WEEO
IfiEON
^'24
24H
EKIE : ?O^L
C R
I
c
H
0
R
0
E
u
I
R
M G
S
N
2) / 0 0
g&?$^ 2/ / 0 0
48H
DME : ^0^1
C R
i
c
H
0
R
0
E u
I
R
M
G
S
N
72H
DME : /^^
CR i
c
H
0
R
0
E
U
I
R
M
G
S
N
A A
A
-'""1 -
^-/
G
0
^
3
0A A 0
^ A0 0
'IDIM/3M3TCE IN3KE'
:
/WDTKL
c
ICKEN
C R I cC R i C H 0 R 0H 0 R 0 E U I RE U I R M G S NM G S N
B %
^
0
^
W
^r ^
diL^i
0,00
^ ^
2
A 0 ^ <0(? 0^ CB Ofr{s(7(
WEBEfB^EEE AHw/ ^ A^A-\ / \^ //4^o3>/ r/--1 CS c<
CBSEE5ffa305 CERCTSES
TETCS
IH3UEE
MJMEEO
IfiEIN
i'
1
-.
/
cla^
/
^
Z
c
R
I c
H
0
R 0
E
U
I R
M
G
S N
y^w
3^^S
7. A y 0
t 1 o 0.
J8
^../oL/<2
c
R
I c
H
0
R 0
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U I
R
-
M
G
S N
00^ 0
<~\ --\
^
^-1
^
J3S
../../..
C
R
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H
0
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U
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M G SN
BaBREHE BELEE
W^/ch
A^^ / 6'S
/
iSiffipany Bafiiti.ee^. ^oes ^ sonfasn T SCA Ct-n
