Document EdnBbX3ymeaELQGyyj81Zgxjn
4) OECD 405-OPPTS 870.2400, Acute eye irritation (rabbit), 132005
Acute Ocular IrritationStudy in New Zealand White Rabbits
Study Number: 132-005
ACUTE OCULAR IRRITATION STUDY OF T-7485 APPLIED TO
NEW ZEALAND WHITE RABBITS
STUDY NUMBER: 132-005
SPONSOR
3M Corporate Toxicology 3M Center, Building 220-2E-02
P.O. Box 33220 St. Paul, MN 55133-3320 Telephone: 651-737-2678
FAX: 651-733-1773
SANlTlZED
DEC 0 9 2003
SPONSOR'S REPRESENTATIVE
John L. Butenhoff, Ph.D., DABT, CIH Senior Laboratory Manager
TESTING FACILITY
Primedica Redfield Mail: P.O. Box 308 Deliveries: 100 East Boone St. Redfield, AR 72132 Telephone: 501-397-2540 FAX: 501-397-2002
STUDYDATES
Study initiation: June 8, 2000 Animal Phase Initiation: June 9, 2000 Animal Phase Completion: June 29, 2000
Study Completion: August 9, 2000
Page 1
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
GLP COMPLIANCE STATEMENT
Study Number: 132-005
I certify that this study was perFormed in compliance with the United States Food and Drug Administration (FDA) Good Laboratory Practice Regulations (21 CFR Part 58)and OECD
Regulations [C(81)30 (final)] and that the report accurately reflects the raw data.
/
Johfi Seng, Ph.D. Study Director Primedica Redfield
Date
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Acute Ocular IrritationStudy in New Zealand White Rabbits
Study Number: 132-005
QUALITY ASSURANCE STATEMENT
Study Number: 132-005
This study has been inspected and audited by the Quality Assurance Unit (QAU) as required by the Good Laboratory Practice (GLP) regulations promulgated by the U.S. Food and Drug Administration and OECD Regulations. The following is a record of the dates that auditdinspections were performed and reported by the QAU.
DATEOF AUDITANSPECTION
I 1 (06/08/00
TYPEOF AUDIT~~NSPECTION
Protocol
I
06/09/00
Dose Administration, Ocular Scoring, and Clinical Observations
DATESREPORTEDTO STUDY DIRECTORAND MANAGEMENT
II 06/08/00
06/09/00
I 07/11/00
07119/00
In-Life Raw Data, Formulations Raw Data, and Tables
Draft Report
07/12/00 07/19/00
08/0 910 0
Final Report
08/09/00
The report accurately reflects the original data APPROVED BY:
Jdlie f;omboli, B/S. Assuradce Auditor
1 Date
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Acute Ocular IrritationStudy in New Zealand White Rabbits
Study Number: 132-005
TITLE
ACUTE OCULAR IRRITATION STUDY OF T-7485 APPLIED TO
NEW ZEALAND WHITE RABBITS
APPROVED BY:
Johh Seng, Ph.D. Study Director Primedica Redfield
m//
U
Trn
Dbte
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
TABLE OF CONTENTS
Paqe
Cover Page....................................................................................................................................... 1 GLP Compliance Statement ............................................................................................................. 2 Quality Assurance Statement ........................................................................................................... 3 Title and Signature Page.................................................................................................................. 4 Table of Contents ............................................................................................................................. 5 Study Abstract .................................................................................................................................. 6 Objective........................................................................................................................................... 7 Materials and Methods ..................................................................................................................... 7 Archival Statement............................................................................................................................ 8 Results.............................................................................................................................................. 9 Conclusion ........................................................................................................................................ 9 Appendices ..................................................................................................................................... 10
Appendix 1 .Individual Clinical Observations.......................................................................... 11
Appendix 2 .Individual Ocular Scores and Ocular Irritation Ratings....................................... 14
Appendix 3 .Certificate of Analysis ......................................................................................... 20 Appendix 4 .Protocol and Amendment ................................................................................... 22 Appendix 5 .Protocol Deviations............................................................................................. 34 Appendix 6 .Key Personnel..................................................................................................... 36
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
ACUTE OCULAR IRRITATION STUDY OF T-7485 APPLIED TO NEW ZEALAND WHITE RABBITS
STUDY ABSTRACT
The objective of this study was to determine the acute effects of a single ocular application of test material to New Zealand White rabbits.
The rabbits were individually housed in stainless steel cages. Animal identification consisted of uniquely numbered ear markings and cage cards. On Study Day 1, the animals were approximately 16 weeks old.
Group Number
Group Designation
Dosage Level (mg)
Number of Animals Females
1
Limit Dose
1oo*
3
I
J
*Due to test material adherence to the weigh boats, approximately 80 mg
of test material was instilled into each animal's left eye.
The test material (approximately 80 mg) was instilled into the left eye of each animal on Study Day 1. Both eyes were held closed for approximately two seconds. Both eyes were flushed with approximately 5 mL of 0.9% saline after the 24-hour score was recorded.
Observations for mortality and moribundity were recorded twice daily (a.m. and p.m.). Clinical observations were recorded predose, approximately one hour postdose, once daily thereafter, and when a change was noted. Predose clinical observations were all normal while at one hour postdose, excessive lacrimation of the left eye was recorded in all three rabbits. The excessive lacrimation of the left eye persisted throughout the remainder of the study. All animals' eyes were normal when the pretest ophthalmologic examinations were performed.
Maximum average ocular scores for 24- and 72-hours postdose were 30 and 35, respectively. By
definition, the findings at 24- and 72-hours will not maintain the categories of non-irritating, practically non-irritating, or minimally irritating. Ocular irritation scores at the 7-day reading were
less than 10 (scoreswere 6, 4, and 30 for the three rabbits) for 67% of the rabbits. Also, the
seven-day mean score was 13. By definition, the resulting scores indicate that T-7485 should be
rated as a moderate irritant.
In conclusion, instillation of approximately 80 mg of T-7485 produced a moderate irritant rating.
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
OBJECTIVE
The objective of this study was to determine the acute effects of a single ocular application of test material to New Zealand White rabbits.
MATERIALS AND METHODS
TEST AND CONTROL MATERIALS: The test material and vehicle were identified as follows:
Test Material: Lot Number: Date Received: Physical Description: Storage: Amount Received:
T-7485 (Potassium PerfluorobutaneSulfonate, PFBS) 2 April 6, 2000 White Powder Room Temperature 500 grams
The Sponsor assumed responsibility for characterization (identity, purity, and stability) determinations of the test material. The test material was inventoriedwhen received at Primedica Redfield, and a record of all test material usage was maintained. The Certificate of Analysis for the test material is in Appendix 3.
The test material was used as received.
TEST ANIMALS: Healthy female New Zealand White rabbits were received from Myrtle's Rabbitry, Inc. for use on the study. The animals were individually housed in stainless steel cages. Animal identification consisted of uniquely numbered ear markings and cage cards. On Study Day 1, the animals were approximately 16 weeks old.
Teklad Certified Rabbit Diet #8630 was provided at approximately 125 grams per day, and filtered tap water was provided ad libitum. The feed and water were routinely analyzed for contaminants. There were no known contaminants in the feed or water that would be expected to affect the results of the study. The results of these analyses are on file at Primedica Redfield.
Group Number
1
Group Designation
Limit Dose
Dosage Level (mg)
1oo*
Number of Animals Females
3
The test material (approximately 80 mg) was instilled into the left eye of each animal on Study
Day 1. Both eyes were held closed for approximately two seconds. Both eyes were flushed with
approximately 5 mL of 0.9% saline after the 24-hour score was recorded.
Page 7
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
JUSTIFICATION FOR SPECIES SELECTION, NUMBER OF ANIMALS, ROUTE OF ADMINISTRATION, AND DOSE LEVEL: Rabbits are an animal model for acute toxicity studies of this type. The number of animals assigned to the study represented the minimum required to meet the objective of the study and the OECD guidelines (M05).
In the assessment and evaluation of the toxic characteristics of a substance, determination of the irritant andlor corrosive effects on the eyes is necessary. Dose levels were chosen by the OECD guideline M05.
CLINICAL OBSERVATIONS: Observations for mortality and moribundity were recorded twice daily (a.m. and p.m.). Clinical observations were recorded predose, approximately one hour postdose, once daily thereafter, and when a change was noted.
OPHTHALMOLOGIC EXAMINATIONS: An ophthalmological examination was performed by the Staff Veterinarian prior to dose administration to verify that all animals' eyes were normal.
OCULAR SCORING: Ocular irritation was scored at approximately 1, 24, 48, and 72 hours postdose and daily on Study Days 5 through 21 using the scoring scale found in the protocol
(Appendix 4). .
STUDY TERMINATION: On Study Day 21, the study was terminated and all animals were euthanized via an overdose of sodium pentobarbital. No necropsies were performed.
STATISTICS: No statistical analyses were performed.
ARCHIVAL STATEMENT
All original data and the original final report will be retained at Primedica Redfield's archives, located at 100 East Boone Street, Redfield, Arkansas, for a period of five years after issuance of the final report. After this time, the Sponsor will be contacted for disposition instructions.
Page 8
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
RESULTS
CLINICAL OBSERVATIONS: Observations for mortality and moribundity were recorded twice daily (am. and p.m.). Clinical observations were recorded predose, approximately one hour postdose, once daily thereafter, and when a change was noted. The individual clinical observations are in Appendix 1.
Predose clinical observations were all normal, while at one hour postdose, excessive lacrimation of the left eye was recorded in all three rabbits. The excessive lacrimation of the left eye persisted throughout the remainder of the study.
OPHTHALMOLOGIC EXAMINATIONS All animals' eyes were normal when the pretest ophthalmologic examinations were performed.
OCULAR SCORING: Due to test material adherence to sample containers, only approximately 80% of the 100 mg of test material was instilled in the left eye of the rabbits. Ocular irritation was scored at approximately 1, 24, 48, and 72 hours postdose and daily on Study Days 5 through 21 using the scoring scale found in the protocol (Appendix 4). The individual ocular scores are located in Appendix 2.
Maximum average ocular scores for 24- and 72-hours postdose were 30 and 35, respectively. By definition, the findings at 24- and 72-hours will not maintain the categories of non-irritating, practically non-irritating, or minimally irritating. Ocular irritation scores at the 7-day reading were less than 10 (scores were 6,4, and 30 for the three rabbits) for 67% of the rabbits. Also, the seven-day mean score was 13. By definition, the resulting scores indicate that T-7485 should be rated as a moderate irritant (as defined in Ocular Irritation Ratings, Appendix 2).
CONCLUSION
In conclusion, instillation of approximately 80 mg of T-7485 produced a moderate irritant rating.
Page 9
Acute Ocular IrritationStudy in New Zealand White Rabbits
APPENDICES
Study Number: 132-005
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
APPENDIX 1 INDIVIDUAL CLINICAL OBSERVATIONS
Page 11
PROTOCOL 132-005: ACUTE OCULAR IRRITATION STUDY OF T - 7 4 8 5 APPLIED TO NEW ZEALAND WHITE RABBITS
A. P. P. .E.N.D.IX. . .1.: . . .T.IM. .E.0. .C.L.I.N.IC. .A.L. .O.B.S.E.R.V.A.T.I.O.N.S. .-. .I.N.D.I.V.ID. .U.A.L. .-. .F.E.M.A.L.E. .R.A.B.B.I.T.S. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
S_TUD_Y D_AY _1 _ _DOSE_GR_OUP_1 _8_0 M_G _ _ _ . _ _ _ _ - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - . - - - - - -
R_AB_BIT_# __PRE_DOS_E ._____1_HOU_R P_OST_DOS_E __-------------------------.--------------------------------
1674
APPEARS NORMAL
EXCESSIVE LACRIMATION, LEFT EYE
1676
APPEARS NORMAL
EXCESSIVE LACRIMATION, LEFT EYE
1-b7_8 ___APP_EAR-S _NORM_AL ____EXC_ESS-IVE_LA_CRIM_AT_ION,_LE_FT E_YE _________________- -__ _-_ --_-_ _-_-_ _-_-_ -_ _-_-_ _-_-_ _-_-_-_ _-_-------
PROTOCOL 132-005: ACUTE OCULAR IRRITATION STUDY OF T-7485 APPLIED TO NEW ZEALAND WHITE RABBITS
A. P. P. .E.N.D.I.X. .1.:. . .C.L.I.N.I.C.A.L. .O.B.S.E.R. V. A. .TI.O.N.S. . .- . .IN. D. .IV.I.D.U.A. L. . -. . F. E. M. .A.L.E. R. .A.B.B.IT. S. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
D.O. S. E. . G. .R.W. .P. .1. . . . . . . . . . . . . . . .8.0. .M.G. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
R. .A.B.B.I.T. .#. . . . . . . . . . . . . . . . . . . . . . .D.E.S.C. R. .IP.T.I.O.N. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1674
SO( 2 - 6 ) SD( 7- 12)
EXCESSIVE LACRIMATION, LEFT EYE LACRIMATION, LEFT EYE
SD( 13- 18)
EXCESSIVE LACRIMATION, LEFT EYE
SD( 20 )
EXCESSIVE LACRIMATION, LEFT EYE
1676 SD( 2- 6 )
EXCESSIVE LACRIMATION, LEFT EYE
SD( 7- 1 2 )
LACRIMATION, LEFT EYE
SD( 13- 20)
EXCESSIVE LACRIMATION, LEFT EYE
1678 SO( 2- 6 )
EXCESSIVE LACRIMATION, LEFT EYE
SD( 7- 12)
LACRIMATION, LEFT EYE
SD( 13- 20)
EXCESSIVE LACRIMATION, LEFT EYE
___S_D( _2_1 _) ___LAC_RIM_ATIO_N, _LE_FT _EYE________________________----------------------------
SD = STUDY DAY
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
APPENDIX 2 INDIVIDUAL OCULAR SCORES AND OCULAR IRRITATION RATINGS
Page 14
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
ANIMAL NUMBER
1674 1676 1678
CORNEA
OPACITY AREA
1
1
1
1
2
3
ANIMAL NUMBER
1674 1676 1678
CORNEA
OPACITY AREA
1
1
2
2
1
1
ANI MAL NUMBER
1674 1676 1678
CORNEA
OPACITY AREA
0
0
2
2
2
3
ANIMAL NUMBER
1674 1676 1678
CORNEA
OPACITY AREA
0
0
2
2
2
3
ANIMAL NUMBER
1674
1676 1678
CORNEA
OPACITY AREA
0
0
1
1
2
3
ANIMAL NUMBER
1674 1676 1678
CORNEA
OPACITY AREA
0
0
1
1
2
3
1 HOUR POSTDOSE
IRIS
CONJUNCTIVAE REDNESS CHEMOSIS DISCHARGE
2
2
4
3
1
2
1
3
2
2
2
3
24 HOURS POSTDOSE
CONJUNCTIVAE IRIS REDNESS CHEMOSIS DISCHARGE
1
2
3
3
1
2
2
3
1
2
2
3
48 HOURS POSTDOSE CONJUNCTIVAE
IRIS REDNESS CHEMOSIS DISCHARGE
1
2
2
3
1
2
2
3
1
2
3
3
72 HOURS POSTDOSE
CONJUNCTIVAE
IRIS REDNESS CHEMOSIS DISCHARGE
1
1
1
3
1
2
2
3
1
2
3
3
STUDY DAY 5
IRIS
0
1 1
REDNESS
1
2 2
CONJUNCTIVAE
CHEMOSIS DISCHARGE
1
2
1
2
3
3
STUDY DAY 6
IRIS
0 0 1
REDNESS 1 2 2
CONJUNCTIVAE
CHEMOSIS DISCHARGE
1
2
1
2
3
3
Page 15
Acute Ocular IrritationStudy in New Zealand White Rabbits
Study Number: 132-005
ANIMAL NUMBER
1674 1676 1678
CORNEA OPACITY AREA
0
0
0
0
1
3
r
ANIMAL NUMBER
1674 1676 1678
CORNEA OPACITY AREA
0
0
0
0
1
3
ANIMAL NUMBER
1674 1676 1678
CORNEA OPACITY AREA
0
0
0
0
1
3
ANIMAL NUMBER
1674 1676 1678
CORNEA OPACITY AREA
0
0
0
0
1
3
ANIMAL NUMBER
1674
1676 1678
CORNEA
OPACITY AREA
0
0
0
0
1
3
ANIMAL NUMBER
1674 1676 1678
CORNEA OPACITY AREA
0
0
0
0
1
3
ANIMAL NUMBER
1674 1676 1678
CORNEA OPACITY AREA
0
0
0
0
2
3
STUDY DAY 7
IRIS
0 0 1
REDNESS
1 1 2
CONJUNCTIVAE CHEMOSIS DISCHARGE
1
1
0
1
2
1
STUDY DAY 8
IRIS
0 0 1
REDNESS
1 1 2
CONJU NCTIVAE
CHEMOSIS DISCHARGE
0
1
0
1
2
1
STUDY DAY 9
IRIS
0 0 I
REDNESS
1 1 2
CONJUNCTIVAE CHEMOSIS DISCHARGE
0
1
0
1
2
1
STUDY DAY 10 CONJUNCTIVAE
IRIS REDNESS CHEMOSIS DISCHARGE
0
0
0
1
0
0
0
1
1
2
2
1
STUDY DAY 11
CONJUNCTIVAE
IRIS REDNESS CHEMOSIS DISCHARGE ,
0
0
0
1
0
0
0
1
1
1
2
1
STUDY DAY 12
IRIS
0 0 1
REDNESS
0 0 1
CONJUNCTIVAE CHEMOSIS DISCHARGE
0
1
0
1
2
1
STUDY DAY 13 CONJUNCTIVAE
IRIS REDNESS CHEMOSIS DISCHARGE
0
0
0
1
0
0
0
1
1
1
3
2
Page 16
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
ANIMAL NUMBER
1674 1676 1678
CORNEA
OPACITY AREA
0
0
0
0
2
4
ANIMAL NUMBER
1674 1676 1678
CORNEA OPACITY AREA
0
0
0
0
2
4
ANIMAL NUMBER
1674 1676 1678
CORNEA OPACITY AREA
0
0
0
0
3
4
ANIMAL NUMBER
1674 1676 1678
CORNEA OPACITY AREA
0
0
0
0
3
4
ANIMAL NUMBER
1674
1676 1678
CORNEA OPACITY AREA
0
0
0
0
3
4
ANIMAL NUMBER
1674 1676 1678
CORNEA
OPACITY AREA
0
0
0
0
4
4
ANIMAL NUMBER
1674 1676 1678
CORNEA OPACITY AREA
0
0
0
0
4
4
STUDY DAY 14
IRIS
0 0 1
REDNESS
0 0 2
CONJUNCTIVAE CHEMOSIS DISCHARGE
0
1
0
1
3
3
STUDY DAY 15
IRIS
0 0 1
REDNESS
0 0 2
CONJUNCTIVAE CHEMOSIS DISCHARGE
0
1
0
1
2
3
STUDY DAY 16
IRIS
0 0 1
REDNESS
0 0 2
CONJUNCTIVAE CHEMOSIS DISCHARGE
0
1
0
1
2
3
STUDY DAY 17 CONJUNCTIVAE
IRIS REDNESS CHEMOSIS DISCHARGE
0
0
0
1
0
0
0
1
1
2
2
3
STUDY DAY 18 CONJUNCTIVAE
IRIS
REDNESS CHEMOSIS DISCHARGE
0
0
0
1
0
0
0
1
1
2
3
1
STUDY DAY 19
IRIS
0 0 2
REDNESS 0 0 2
CONJUNCTIVAE
CHEMOSIS DISCHARGE
0
0
0
1
3
1
STUDY DAY 20
IRIS
0 0 2
REDNESS
0 0 2
CONJUNCTIVAE CHEMOSIS DISCHARGE
0
1
0
1
3
1
Page 17
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
ANI MAL NUMBER
1674 1676 1678
CORNEA
OPACITY AREA
0
0
0
0
4
4
STUDY DAY 21
IRIS
0 0 2
REDNESS
0 0 2
CONJUNCTIVAE
CHEMOSIS DISCHARGE
0
0
0
0
3
1
Page 18
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
APPENDIX 2 OCULAR IRRITATION RATINGS
Rating Non-l rritating Practically Non-Irritating
~~
Min imally Irritating Mildly Irritating Moderately Irritating
Severely Irritating
Extremely Irritating
Maximum Average Score
0.00 - 0.50
0.51 - 2.50
2.51 - 15.00
15.01 - 25.00
25.01 - 50.00
Definition
To maintain this category, all scores at the 24-hoF reading must be zero; otherwise, increase category one level. To maintain this category, all scores at the 24-hour reading must be zero: otherwise, increase category one level. To maintain this category, all scores at the 72-hour reading must be zero; otherwise, increase category one level. To maintain this category, all scores at the 7-day reading must be zero; otherwise, increase category one level. To maintain this category, scores at the 7-day reading must be less than or equal to 10 for 60% or more of the animals. Also the 7-day mean score must be less than or equal to 20. If the 7-day mean scores are less than or equal to 20, but less than 60% of the animals show scores less than 10, than no animal among those showing scores greater than 10 can exceed a score of 30 if the category is to be maintained; otherwise, increase category one level.
50.01 - 80.00 80.01 - 110.00
To maintain this category, scores at the 7-day reading must be less than or equal to 30 for 60% or more of the animals. Also the -/-day mean score must be less than or equal to 40. If the 7-day mean scores are less than or equal to 40, but less ihan 60% of the animals show scores less than 30, than no animal among those showing scores jreater than 30 can exceed a score of 60 if the :ategory is to be maintained; otherwise, increase
2ateaor-v one level.
NOTE: The category of the test material is not to be increased more than one level above its maximum average score.
Page 19
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
APPENDIX 3 CERTIFICATE OF ANALYSIS
Page 20
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
Certificate Of Analysis
CJ?$O3X+ (PFBS), Lot 2
March 10,2000
I
CJF~S~X'
ca9s03x'
0.13 % 99.82 % 0.04 %
Additionally, the isomer distribution of the sample was determined using contain the following weight percent composition:
.techniques and found to
97.86 %
1.96 %
Page 21
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
APPENDIX 4 PROTOCOL AND AMENDMENT
Page 22
~~~
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
Primedica Redfield Protocol
~~~
Prorocol Number: 132oM
ACUTE OCULAR IRRITATIONSTUDY OF T-7485 APPLIED TO NEW ZEALAND WHITE RABBITS
PROTOCOL NO.: 132005
OBJECTNE: Determinethe acute effect(s) of a single Ocular application of test material to New Zeabnd White rabbits.
LOCATION OF STUDY AND CONDITIONS OF TESTING:
It is the intention of 3M Corporate Toxicology, through the conduct of this study, to generate animal saw
data that may be submitted to regulatory authorities. Primedica Redfield. 100 East Boone Street, Redfield, Arkansas, 72132, is accredited by AAALAC and licensed by the United States Departmentof Agriculture to conduct research in laboratory animals. All the conditions of testing will conform to the Animal Welfare Act (CFR 9) and its amendments. Primedica Redfield will follow all requirements specmed in this approved protocol and all applicable governmental regulations regarding Good Laboratory Practices as well as Primedica Redfields Standard Operating Procedures. Changes in the protcml may be made by consultationwith and approval from 3M Corporate Toxicology followed by written verification of the change. 9M CorporateToxicology reservesthe right to inspect facilities and procedures usedfor this study by means of announced or unannounced site visits. Primedica Redfieldwill notify 3M C o m Toxicology promptly by telephone prior to release of any data for review.
SPONSOR:
3M Corporate Toxicology 3M Center, Building 220-2E-02 P.O. Box 33220
St Paul, M N 55133-3320
Telephone: 651-737-2678
FAX: 651-733-1773
SPONSOR'S REPRESENTATWE:
John L. Butenhoff, Ph.D. DABT, CIH Senior Laboratory Manager
TESTING FACILITY:
Primedica Redfield Mail: P.O. Box 308 Deliveries: 100 East Boone St. Redfield, AR 72132 Telephone: 501-397-2540 FAX: 501-397-2002
PERSONNEL:
Study Director: John Seng, Ph.D.
Principle Investigator: KarenTranter, B.A., U T G Veterinarian: Allan Manus, D.V.M.. M S c . . ACLAM Pathologist TBD
PROPOSED STUDY DATES:
Experimental Start Date: June 9, 2000 Experimental Termination Date: TBD DraR Report Date: July 24, 2000
Page I of 10
06/08/00
Page 23
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
Primedica Redfield Protocol
Prorocol Number: 132405
1.
REGULATORCYOMPLIANCEAND QUALTIY ASSURANCE
This study will be conducted in accordance with the following Good Laboratory Practice RegulationslStandards/Guidelines:
R 21 CFR 58 R C(81)30 (Final) (OECD)
The Quality Assurance Unit, in accordancewith Primedica Redfields Standard Operating Procedures(SOPS).will audit the protocol, study conduct, and the final report.
2.
INSTITUTIONAL ANIMAL CAREAND USE COMMITTEE& V I W
The protocol will be reviewed and approved by Primedica Redfield's InstitutionalAnimal Care and Use Committee(IACUC)for compliance with regulations prior to study initiation.
In the opinion of the Sponsor, indicated by the signature on this protocol, the study does not unnecessarilyduplicate any previouswork
3. MATERIALS
TESTMATERIAL
T-7485 (Potassiumpertluorobutanesulfonate, PFBS)
/DEN7JF/CATION
The lot number will be listed in the raw data and final report.
PHYSICADLESCRlPTION White powder
STORAGE CONDITIONS
The bulk test material will be stored in its original container and will be stored at room temperature. The test material will be used as received.
ANALMlCAL CHEMISTRY
The Sponsor assumes responsibility for characterization (idenbty, purity, and stability) of the bulk test material (including under test conditions). Infomation on the composition and method of synthesis of the bulk test materialwill be held by the Sponsor. The doses, unless otherwise stated, will be calculated assuming the test material to be 100% pure.
INVENTORY
The test material container@)will be inventoried when received at the testing laboratory, and a record of all test material use will be maintained. The technician dosing the animalswill: I )Record the weight or volume of the test material container immediately before and after dosing;
2) Compute and record the amount used and compare it to the theoretical
amount immediately after dosing; 3) Keep the formulated test material
use log in the animal study books at all times; 4) The dose used must be within ?IO% of theoretical.
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
Primedica Redfield Protocol
ProiocolNumber: 132m5
TESTMATERIAL RETENTION
Unused test material may be returned to the Sponsor or designee at the
termination of the study, or retained for use on future studies. The Sponsor will be notified in advance of shipping, and a transmittal letter will
accompany the shipment. The material will be packed in a suitable
container to maintain the conditions specified by the Sponsor during transit plus an adequate margin of safety for any transit delays.
SAFETY PRECAUTIONS
4.
ANIMALS
General safety precautions as required by Primedica Redfields policies
and procedures will be followed. The Sponsofs Representativewillbe notified of any personnel exposures requiring a physician's examination
or care.
Species: StrainlSource:
Age at Initiation: Number and Sex:
Identification:
Rabbits New Zealand white, Myrtle's Rabbitry, Thompson Station, Tennessee At least 12 weeks Three females (female nulliparous and non-
pregnant) Indelible ink and colorcoded cage card
ANIMAL HUSBANDRY
HOUSING
FEED
The animals will be individually housed in stainless steel cages. The cages conform to standards set forth in the Guide for the Care and Use of Laboratow Animals, National Academy Press, Washington, D.C., 1996.
Teklad Certified Rabbit Diet #8630 will be provided at approximately 125 grams per day. This diet is routinely analyzed by the manufacturer for nutritional components and environmental contaminants, Results of the manufacturer's analyses are on file at Primedica Redfield.
WATER
Filtered tap water will be provided adlibitum. Samples of the water are
analyzed for total dissolved solids, hardness, and specified microbiologicalcontent and for environmental contaminants. Results of these analyses are on file at Primedica Redfield.
CONTAMINANTS
There are no known contaminants in the feed or water that would be expected to interfere with this study.
ENVIRONMENT
Environmental controls are set to maintain a temperature of 16' to 22% (61' to 72F) with a relative humidity of 30% to 70%. These parameters are recorded at least once daily. A 12:12 hour lightdark cycle is maintained.
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
Primedica Redfield Protocol
Protocol Number: 132005
Animals will be acclimated for a minimum of seven days prior to the study
start. An adequate number of extra animalswill be purchasedso that no animal in obviously poor health is placed on test. At the request of the Study Director, excess animals may be used as replacement animals. Animals will be examined by the Staff Veterinarian prior to release of that shipment of animals for use on the study.
JUSTIFICATION FOR SPECIES SELECTIONAND NUMBEROF ANIMALS
Rabbits are an animal model for acute toxicity studies of this type. The number of animals assigned to this study represents the minimum required to meet the objedive(s) of the study and the OECD guidelines (M05).
5.
EXPERIMENTAL METHODOLOGY
STUDYDESIGN
Group Number
Group Designation
Dosage Level (m9)
Number of Animals Females
FREQUENCYAND DURATION OF ADMINISTRATION
One hundred milligrams of test material will be instilled into the left eye of each animal on Study Day 1. Both eyes will be held shut for approximately two seconds. Both eyes will be flushed with 0.9% saline
after the 24 hour score has been recorded (SOP252.07).
JUSTIFICATION FOR ROUE OF ADMINISTRATION AND DOSELEVEL
In the assessment and evaluation of the toxic characteristics of a
substance, determination of the irritant andor corrosive effects on the eyes of mammals. Dose levels were chosen by the OECD guideline #405.
ANTEMORTEMOBSERVATIONS
CLINICAL OBSERVATIONS
Observations will be performed and recorded twice daily (a.m. and p.m.)
for moribundity and mortality.
Clinical observations will be performed, according to Primedica Redfields SOPS, and recorded predose. approximately o n e hour postdose. once daily thereafter, and when a change is noted.
oPKTHALMOLCGlC .&4MlNATlONS
An ophthalmological examination will be conducted by a staff veterinarian within 24 hours before testing to verify that the eyes are normal.
-
~~
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
Primedica Redfield Protocol
Prorocol Number: 132-005
OCULAR SCORING
Ocular irritation will be scored at approximately 1,24,48. and 72 hours postdose, as defined in AttachmentA. Ifthere is no evidence of irritation at 72 hours postdose, the study may be terminated. Extended observationmay be necessary if there is persistent corneal involvement or other Ocular irritation in order to determine the progress of the lesions and their reversibility or irreversibility.
POSTMORTEM OBSERVATlONS
MORISUND ANIMALSAND ANIMLSFOUNDDEAD
Animals unlikely to survive until the next scheduled observation will be weighed, euthanized, and necropsied. Animals found dead will be weighed and necropsied. In either case, a complete necropsy will be performed as detailed below. Euthanasia of moribund animals will be authorized by the Study Director or Staff Veterinarian with the concurrence of the Study Director.
All animals surviving to the end of the study may be returned to the testing facility's methodology population.
NECROPSY
All animals that die during the study will be subjected to a complete necropsy examination. A complete necropsy is defined as examination of the external surface of the body, all orifices, and the cranial, thoracic, and abdominal cavities, and their contents. Gross lesions will be preserved in a suitable fixative for possible histopathology; all other tissues will be discarded. Histopathology will be provided at additional expense to the Sponsor.
6.
STAllSnCAL ANALYSS
Statistical analysis will not be required.
7.
REPORT
An audited draft report will be sent to the Sponsor. Revisions to the initial draft report will be provided to the Sponsor by mail or fax transmission as printed copies of the corrected pages only. Additional revisions or complete copies of the revised draft reports will be provided at additional expense to the Sponsor.
Three copies (one bound, two unbound) of the approved final report will be submitted to the Sponsor approximately two weeks after approval of the draft report. The final report will include all elements requiredhecommended by the regulations and guidelines.
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
Primedica RedfieldProtocol
Protocol Number: 132-035
The report will include. but is not limited to, those items listed below:
Descriptive text of the study objective Summary Test material identilication (Certificate of Analysis, stability data, and analytical analysis if any are performed) Methods Results and conclusions Ocular scoring A copy of the protocol, all protocol amendments, and all protocol deviations that may affect the integrity of the study
8.
MAINTENANCE OF RAW DATA AND RECORDS
Original data or copies thereof, will be available at Primedica Redfield to facilitate auditing the study during its progress and before acceptanceof the final report When the final report is completed. all original paper data and the original final report will be retained in the archives of the laboratory for at least five years. Wet tissues, slides, and blocks (if generated) will be retained at Primedica Redfield for one year. After one year the storage of the wet tissues, slides, and blocks will be negotiated with the Sponsor. After five years the storage of the paper data will be negotiatedwith the Sponsor. The Sponsor will be notified prior to disposal of any original study data.
9.
QIJAUTYASSUR~NCRE VIEW
This is to certify that this protocol has been reviewed by Quality Assurance.
Val Gartner, B.A Quality Assurance Auditor Primedica Redfield
IO. APPROVALS
ipldm Date
&e,& 7- e& +?a
' John L. Butenhoff, Ph.D, DABT, CIH/
6/~,/$4
' Date'
Senior Laboratory Manager
3M Corporate Toxicology
LAW(/
SJ'btuhb),ySDeinregc. tPohr.D.
U
Primedica Redfield
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
Primedica Redfield Protocol
Prorocol Number: 132-005
ATTACHMENT A OCULAR IRRITATION SCORING SYSTEM
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06lQB/OO
Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
Primedica Redfield Protocol
Protocol Number: 132-005
ARACHMENT A OCULAR IRRITATION SCORING SYSTEM
CORNEA
GRADT
OpacW degree of density (most dense area used for reading)
No Ulceration or Opacity ....................................................................................................... 0
Scattered or diffuse areas of opacity (other than slight dulling of normal luster), details of iris
clearly visible ............................................................................................................... 1'
Easily discernible translucent area, details of iris slightly obscured
................. 2'
Nacreous area, no details of iris visible, sue of pupil barely discernible ..............................
' 3
Opaque cornea, iris not discernible through the opacity .......................................................
4'
Normal ..................................................................................................................................
o
Markedly deepened rugae (folds), congestion, swelling, moderate circumcorneal hyperemia
or injection (any of these or any combination of these), iris still reacting to light
(sluggish reaction is positive) ......................................................................................
' 1
No reaction to light, hemorrhage, gross destruction (any or all of these) .............................
2'
CONJUNCTIVAE
GRADE
A. Redness: refers to palpebral and bulbar conjunctivae excluding cornea and iris
Normal Blood Vessels ...............
Some blood vessels definitely hy
Diffuse. crimson color. individual
Diffuse, beefy red .......................
........................................................... 3'
B. Chemosis: swelling of the lids and/or nictitating membrane
No swelling ............................................................................................................................ 0
Any swelling above normal (includes nictitatingmembrane) .
Obvious swelling with partialeversion of the lids ...................
................... 2'
Swelling with lids half closed .................................................................................................
3'
Swellingwith lids more than half closed ...............................................................................
4'
~~~
~
~~
The total score for the eve is the sum of allscores obtained for the cornea. ins. and coniunctivae.
INDICATESA POSITIVEREACTION(Fedeal HazardousSubstance Act Interpretation)
REFERENCES
' Hackett, R.B., and McDonald, T.O., Eye Irritation, Dermatotoxicoloay, Fourth Edition, Marzulli, F.N. and
Maibach, H.I.: 749-815, 1991.
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
Primedica Redfield Protocol
ATTACHMENT A OCULAR IRRITATION SCORING SYSTEM
proroCol Number: 132-005
CORNEA
A. Opacw. degree of density (most dense area used for reading)
No Ulceration or Opacity ......................................................................
Scattered or diffuse areas of opacity (other than slight dulling of norm
clearly visible .............................................................
Easily discernible translucent area, details of ins slightly ob Nacreous area, no details of iris visible, size of pupil barely
..................................... ible .........................
Opaque cornea, ins not discernible through the opacity .........
B. Area Involved: total area exhibiting any opacity regardless of degree
0% . - No Ulceration or Opacity ............................
.......................................
- 025% One quarter or less, but not zero .................................
2550% - Greater than one quarter, but less than half ............................
- 50-75% Greater than half, but less than three quart
75100%- Greater than three quartets up to the whole area ............
C. Fluorescein Staining appearance of staining
Cornea not examined ....................................................................... -No abnormal fluorescein staining .................. Fluorescein staining ............................................................
......................
- Area Involved
~ 2 5 % - One quarter or less, but not zero ..................
................................
2550% - Greater than one quarter, but less than half ..................................................
50-75% Greater than half, but less than three quarters ..............................................
75100%- Greater than three quarters up to the whole area .........................................
A x B x 5
TOTAL MAXIMUM = 80
VALUE
2' 0
A
B
C D
A. Grades
Normal ..................................................................................................................................
0
Markedly deepened rugae (folds), congestion, swelling, moderate circumcorneal hyperemia
or injection (any of these or any combination of these), ins still reacting to light
(sluggish reaction is positive) ......................................................................................
1'
No reaction to light, hemorrhage, gross destruction (any or all of these) .............................
2'
Ax5
TOTAL MAXIMUM = 10
INDICATESA POSITIVE REACTION(FHSA Interpretation) Page 9 of 10
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
Primedica Redfield Protocol
ATTACHMENT A
OCULAR IRRITATIONSCORING SYSTEM (continued)
Protocol Number: 132M)5
CONJUNCTIVAE
VALUE
A. Redness: refers to palpebral and bulbar conjunctivae excluding cornea and iris
Normal Blood Vessels ...............................................................
.................................
0
Some blood vessels definitely hyperemic (injected) ..........................
.........................
1
Diffuse, crimson color, individual vessels not easily discernible ...........................................
2'
Diffuse, beefy red ...............................................................
........................................
3'
B. Chemosis: swelling of the lids and/or nictitatingmembrane
....................................................... 0
ne) ................................................ 1
Obvious swelling with partial eversion of the lids .................................................................. 2'
Swelling with lids half closed .............
................................................
3'
Swelling with lids more than half closed ...............................................................................
4'
C. Discharge
No discharge .........................................................................................................................
0
Any amount different than normal (does not indude small amounts often observed
in the inner canthus of normal animals) ...................................................................... 1
Dischargewith moistening of the lids and hairsjust adjacent to the lids .
2
Discharge with moistening of the lids and hairs, and a considerable area around the eye .. 3
(A + B + C) x 2
TOTAL MAXIMUM = 20
The total score for the eye is the sum of all scores obtained for the cornea, iris, and conjunctivae. * INDICATES A POSITIVE REACTION (FHSA Interpretation)
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
\
Primedica Redfield Protocol Amendment
Pro!mol Numbrr: 132-005
STUDY NUMBER: AMENDMENT: STUDY TITLE:
DATE ISSUED:
132-005 1
ACUTE OCULAR IRRITATIONSTUDY OF T-7485 APPLIED TO NEWZEALAND
WHITE RABBITS July 24,2000
AMENDMENT # 1
ITEM 1: REASON:
Page 8, Attachment A
DELETE
Attachement A contained two different scoring scales. The scale on pages 9 and 10 was used on this study.
APPROVALS:
John L. Butenhoff, Ph.D., DABT, CIH
Senior Laboratory Manager 3M Corporate Toxicology
I
Date
John Sena. Ph.D.
Study Director Primedim Redfield
Date
1
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
APPENDIX 5 PROTOCOL DEVIATIONS
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Acute Ocular IrritationStudy in New Zealand White Rabbits
Study Number: 132-005
PROTOCOL DEVIATIONS There were no protocol deviations that adversely affected the integrity of the study.
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Acute Ocular IrritationStudv in New Zealand White Rabbits
~~
Studv Number: 132-005
APPENDIX 6 KEY PERSONNEL
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Acute Ocular Irritation Study in New Zealand White Rabbits
Study Number: 132-005
KEY PERSONNEL Study Number: 132-005
Study Director: ............................................... Study Coordinator:......................................... Head Technician............................................ Veterinarian: .................................................. Quality Assurance: ......................................... Report Coordination: ..................................... Report Preparation:.......................................
John Seng, Ph.D. Karen Tranter, B.A., LATG Catherine Morris, ALAT Allan Manus, D.V.M., M.Sc., ACLAM Julie Tornboli, B.S. Amy Babb, B.S. Tarnmy Peckharn, Ad. Asst. Tammie Hughey, Ad. Asst.
Page 37