Document B8MvaGk47agwy6K8bagxz9GOw
2) OECD 402-OPPTS 870.1200, Acute dermal toxicity (rat), 132010
Acute Dermal Toxicity Study in Spraque-Dawley Rats
Study Number: 132-010
i ACUTE DERMAL TOXICITY STUDY OF T-7485 APPLIED TO SPRAGUE-DAWLEY RATS
STUDY NUMBER: 132-010
SANITIZED
SPONSOR
3M Corporate Toxicology 3 M Center, Building220-2E-02
P.O. Box 33220 St. Paul, MN 55133-3320
SPONSOR'S REPRESENTATIVE
Ph.D., DABT, CIH Senior Laboratory Manager Telephone:
FAX:
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
STUDY DATES
Study Initiation: July 26, 2000 Animal Phase Initiation: July 27, 2000 Animal Phase Completion: August 11, 2000 Study Completion: October 10, 2000
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
GLP COMPLIANCE STATEMENT
Study Number: 132-010
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.
Primedica Redfield
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Acute Dermal Toxicity Study in Spraque-Dawlev Rats
Study Number: 132-010
QUALITY ASSURANCE STATEMENT
Study Number: 132-010
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 auditslinspectionswere performed and reported by the QAU.
DATEOF AUDIT/~NSPECTION
TYPE OF AUDIT/~NSPECTION DATESREPORTETDO STUDY DIRECTORAND MANAGEMENT
07/26/00
ProtocoI
07/26/00
07/27/00
~~
08/10/00.
r08/17/00-08/18/00 0811 8/00
Dose Administration
~
Necropsy
In-Life Raw Data
~
Formulations Raw Data, Report Tables, IndividualAnimal Necropsy Records, and IndividualAnimal Necropsy Observation Tables
07127/00 08/10100 0811 8/00
0811 8/00
09/05/00
Draft Report
09/05/00
10/10/00
Final Report
10/10/00
The report accurately reflects the original data. APPROVED BY:
Val Gartner, B.A. Quality Assurance Auditor Primedica Redfield
di1 u h j ~ Date
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
TITLE
ACUTE DERMAL TOXICITY STUDY OF T-7485 APPLIED TO SPRAGUE-DAWLEY RATS
APPROVED BY:
JSotut-b$ySDeinregc, tPohr .D. Prirnedica Redfield
* ?, /I W
/d /&GO
' Date
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
TABLE OF CONTENTS
Page
Cover Page....................................................................................................................................... 1
GLP Compliance Statement ............................................................................................................. 2
Quality Assurance Statement ........................................................................................................... 3
Title and Signature
........................................................................................................ 4
Table of Contents ...
........................................................................................................ 5
Study Abstract .................................................................................................................................. 6
Objective........................................................................................................................................... 7
Materials and Methods ..................................................................................................................... 7
Archival Statement............................................................................................................................ 8
Results.............................................................................................................................................. 9
Conclusion ........................................................................................................................................ 9
Tables ............................................................................................................................................. 10
Table 1 - Summary of Clinical Observations............................................................................ 11 Table 2 - Summary of Body Weights ....................................................................................... 14 Table 3 - Summary of Body Weight Changes.......................................................................... 17
Appendices ..................................................................................................................................... 20
Appendix 1 - Individual Necropsy Observations .............................. :....................................... 21
Appendix 2 - IndividualClinical Observations.......................................................................... 23 Appendix 3 - Individual Body Weights ..................................................................................... 28 Appendix 4 - Certificate of Analysis ......................................................................................... 31 Appendix 5 - Protocol............................................................................................................... 33
Appendix 6 - Protocol Deviations............................................................................................. 41
Appendix 7 - Key Personnel...................................................................................................... 43
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
ACUTE DERMAL TOXICITY STUDY OF T-7485 APPLIED TO SPRAGUE-DAWLEY RATS
STUDY ABSTRACT
The objective of this study was to determine the acute effect(s) of a single dermal application of T-7485 to Sprague-Dawley rats with a fourteen-day recovery.
Healthy male and female rats were individually housed in stainless steel cages. The animals were acclimated for a minimum of seven days prior to Study Day 1. During the acclimation period, the animals were habituated to collars used during the study. The study design was as follows:
Group Number
1 2 3
Group Designation
High-dose Lowdose
Mid-dose 1
Dosage Level
(mg/kg) 2000 500 1000
Number of Animals*
Males
Females
5
5
5
5
I5 I5
A single dosage level of 2000 mg/kg of test material was applied to the Group 1 animals on Study Day 1. The test material was applied uniformly over an area that was approximately 10% of the body surface area. The test material was held in contact with the skin with a porous gauze dressing and non-irritating tape for a 24-hour exposure period. An Elizabethan collar was placed on each animal during the exposure period. After 24 hours, the test material was removed with warm water and gauze.
Observations for mortality and moribundity were recorded twice daily (a.m. and p.m.). Clinical observations were recorded predose and approximately hourly for four hours postdose on Study Day 1 and then once daily thereafter for at least 14 days. Treatment with T-7485 produced no test material-relatedmortalities, but there was one proceduraldeath that resulted in the replacement of a male rat. Clinical observations included red material around the eyes, nose, and mouth. These observationswere not present after Study Day 2 and were not considered test material-related.
Body weights were recorded pretest and on Study Days 1, 8, and 15. Treatment with T-7485 produced no adverse effects on body weights or body weight changes. When compared to Study Day 1 , Study Days 8 and 15 male mean body weights were increased 17% and 40%,
respectively. Likewise, female mean body weights were increased 8% and 19%, respectively.
On Study Days 1 through 15, male mean body weight change was increased 96.6 grams while body weight change in the females was increased 31.6 granis.
On Study Day 15, all animals (non-fasted)were humanely euthanized via carbon dioxide asphyxiation and submitted for a complete necropsy examination. There were no adverse gross findings recorded for either male or female rats treated with 2000 mg/kg T-7485 with the exception of one male (this male was replaced)which died due to proceduralcomplications.
Treatment with 2000 mg/kg T-7485 produced no treatment-relatedadverse clinical observations, mortality, changes in body weight, or gross pathology findings.
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
OBJECTIVE
The objective of this study was to determine the acute effect(s) of a single dermal application of T-7485 to Sprague-Dawley rats with a fourteen-day recovery.
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, Protected From Light 500 grams
Vehicle:
Supplier:
Lot Number: .
Carboxymethylcellulose(medium viscosity) Sigma Chemical Co. 69H0028
The Sponsor assumed responsibility for characterization (identity, purity, and stability) determinationsof 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 4.
Doses were prepared by Primedica Redfield. The vehicle was prepared to a concentration of 1% by mixing 10 grams of powdered medium viscosity carboxymethylcellulosewith deionized water. The test material was moistened with the vehicle prior to administration to the animals.
TEST ANIMALS: Healthy male and female Crl:CD@(SDI)GS BR stock albino rats were received
from Charles River Laboratories, Inc., for use on the study. The animals were individually housed in stainless steel cages. Animal identificationconsisted of uniquely numbered ear tags and cage cards. On Study Day I,the animals were approximately eight weeks old. The males weighed 233 to 248 grams, and the females weighed 165 to 170 grams.
Teklad Certified Rodent Diet #8728 and filtered tap water were 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.
Environmentalcontrols were set to maintain temperatures of 18" to 26C (64" to 79F) with a relative humidity of 30% to 70%. These parameterswere recorded at least once daily. A 12:12 hour 1ight:dark cycle and ten or greater air changes per hour were maintained in the animal room.
GROUP DESIGNATIONAND TREATMENT: The animals were acclimated for a minimum of seven days prior to Study Day 1 and were examined by the Staff Veterinarian prior to being released for use on the study. During the acclimation period, the animals were habituated to collars used during the study. The study design was as follows:
Group
2 3
Group
Low-dose Mid-dose
I I I Dosage I NumberofAnimals I
Level
Males
Females
-
I 500
5
5
1000
5
5
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
Hair was removed by clipping the dorsal area of the trunk of all animals at least one hour prior to test material administration and as needed thereafter. The dose site and an area of adjacent untreated skin were marked at the corners with an indelible marker.
A single dosage level of 2000 mg/kg of test material was applied to the Group 1 animals on Study Day 1. The test material was applied uniformly over an area that was approximately 10% of the body surface area. The test material was held in contact with the skin with a porous gauze dressing and non-irritating tape for a 24-hour exposure period. An Elizabethan collar was placed on each animal during the exposure period. After 24 hours, the test material was removed with warm water and gauze.
Because there were no test material-related deaths in Group 1, Groups 2 and 3 were not dosed.
JUSTIFICATION FOR SPECIES SELECTION, ROUTE OF ADMINISTRATION, AND DOSE LEVEL: Rats 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 objectives of the study and the OECD guidelines (W02). In the assessment and evaluation of the toxic characteristicsof a substance, determination of acute dermal toxicity is useful where exposure by the dermal route is likely. Dose levels were chosen in accordance with OECD guideline W02.
CLINICAL OBSERVATIONS: Observations for mortality and moribundity were recorded twice daily (a.m. and p.m.). Clinical observationswere recorded predose and approximately hourly for four hours postdose on Study Day 1 and then once daily thereafter for at least 14 days.
BODY WEIGHTS: Body weights were recorded pretest and on Study Days 1, 8, and 15. The pretest body weights are not included in this report but are located in the raw data.
NECROPSY: Animals that died prior to scheduled termination were weighed and necropsied as detailed below. On Study Day 15, all animals (non-fasted)were humanely euthanized via carbon dioxide asphyxiation and submitted for a complete necropsy examination. A complete necropsy was defined as examination of the external surface of the body; all orifices; and the cranial, thoracic, and abdominal cavities and their contents. Gross lesions were preserved in neutral buffered formalin. All other tissues were discarded.
HISTOPATHOLOGY: No histopathologicalexaminations were performed.
STATISTICS: Means and standard deviations were calculated for all quantitative data.
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. Wet tissues, slides, and blocks (if generated) will be retained at Primedica Redfield for one year. After this time, the Sponsor will be contacted for disposition instructions.
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
RESULTS
CLINICAL OBSERVATIONS: The summary of clinical observations is in Table 1. The individual observations are in Appendix 2.
Treatment with T-7485 produced no test material-related mortalities, but there was one procedural death that resulted in the replacement of a male rat. Clinical observations included red material around the eyes, nose, and mouth. These observations were not present after Study Day 2 and were not considered test material-related.
BODY WEIGHTS AND BODY WEIGHT CHANGES: The summaries of body weights and body weight changes are in Tables 2 and 3, respectively. The individual body weights are in Appendix 3.
Treatment with T-7485 produced no adverse effects on body weights or body weight changes. When compared to Study Day 1, Study Days 8 and 15 male mean body weights were increased 17% and 40%, respectively. Likewise, female mean body weights were increased 8% and 19%, respectively. On Study Days 1 through 15, male mean body weight change was increased 96.6 grams while body weight change in the females was increased 31.6 grams.
NECROPSY: The individual necropsy observations are located in Appendix 1.
There were no adverse gross findings recorded for either male or female rats treated with 2000 mglkg T-7485 with the exception of one male (this male was replaced) which died due to procedural complications.
CONCLUSION
Treatment with 2000 mglkg T-7485 produced no treatment-relatedadverse clinical observations, mortality, changes in body weight, or gross pathology findings.
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
TABLES
Study Number: 132-010
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
TABLE 1
SUMMARY OF CLINICAL OBSERVATIONS
Page 11
APPEARS NORMAL, PREOOSE APPEARS NORMAL, 1 HOUR POSTDOSE APPEARS NORMAL, 2 HOURS POSTDOSE APPEARS NORMAL, 3 HOURS POSTDOSE RED MATERIAL AROUND EYES
RED MATERIAL AROUND EYES, 4 HOURS POSTDOSE
RED MATERIAL AROUND NOSE RED MATERIAL AROUNO NOSE, 1 HOUR POSTDOSE RED MATERIAL AROUND NOSE, 2 HOURS POSTDOSE RED MATERIAL AROUND NOSE, 3 HOURS POSTDOSE
RED MATERIAL AROUND NOSE, 4 HOURS POSTDOSE
RED MATERIAL AROUND W T H
N/N = NUMBER OF OBSERVATIONS/NUHBER OF ANIMALS.
5/ 5 2/ 2 21 2 2/ 2
3/ 3 3/ 3 2/ 2
3f 3 3/ 3 3/ 3 5/ 5 1/ 1
APPEARS NORMAL, PREDOSE
5/ 5
APPEARS NORMAL, 1 HOUR POSTDOSE
2/ 2
APPEARS NORMAL, 2 HOURS POSTDOSE
1/ 1
APPEARS NORMAL, 3 HOURS POSTDOSE
I/ 1
RED MATERIAL AROUND EYES
3/ 3
RED MATERIAL AROUND EYES, 1 HOUR POSTDOSE
1/ 1
RED MATERIAL AROUND EYES, 2 HOURS POSTDOSE
1/ 1
RED MATERIAL AROUND EYES, 3 HOURS POSTDOSE
1/ 1
REO MATERIAL AROUND EYES, 4 HOURS POSTDOSE
2/ 2
RED MATERIAL AROUND NOSE
3/ 3
RED MATERIAL AROUND NOSE, 1 HOUR POSTDOSE
2/ 2
RED MATERIAL ARWND NOSE, 2 HOURS POSTDOSE
3/ 3
RED MATERIAL AROUND NOSE, 3 HOURS POSTDOSE
3/ 3
RED MATERIAL AROUND NOSE, 4 HOURS POSTDOSE
5/ 5
R_ED -MA-TE_RIA-L A_ROU-ND-M_OUT_H _ _ _ _ _ . _ - - _ - - - - -1-/ -1- - - - - - - - - - - - - - - - - - - - - - - - - . - - - - - - - - - - - - - - - - - - - - - -
N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF ANIMALS U I T H THE OBSERVATION.
Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
TABLE 2 SUMMARY OF BODY WEIGHTS
Page 14
BOOY UEICHT (G)
SD
1
MEANfS .D .
240.0 f 5.8
SD
8
l4EANfS.D.
281.0 f 6.0
S_D _1_5 ____M_EAN_fS.D_. ___336_.6 f_21_.6 ________________________--------.------------
SD = STUDY DAY
BODY WEIGHT (G)
SD 1
SD
8
MEANfS.D . MEANfS.D .
167.6 f 2.1 181.6 f 5.7
Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
TABLE 3 SUMMARY OF BODY WEIGHT CHANGES
Page 17
-0
11) (P
(D
S_D _ _1 -_15_ _ _ _MEA_NfS_.D._ _ _ _+31_.6 _f _2.2_ _ _ - - - - - - - - - - - - - - - - - - - - - - - - - . - - - - - - - - - - - - - - - - - - - . - - - - - - - - - - - - - - -
SD = STUDY DAY
Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
APPENDICES
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
APPENDIX 1 INDIVIDUAL NECROPSY OBSERVATIONS
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
APPENDIX 1 INDIVIDUAL NECROPSY OBSERVATIONS
GROUP 1 - 2000 mglkg
Animal #6981 -
Animal #6983 -
Male Male
Animal #6985 Animal #6991 Animal #6993 Animal #I6995 Animal #6988 Animal it6990 Animal #6992 Animal #6994 Animal #6996
Male Male Male Male Female Female Female Female Female
No adverse findings. No adverse findings. Died Following Post-Dose Procedure. No adverse findings. No adverse findings. No adverse findings. No adverse findings. No adverse findings. No adverse findings. No adverse findings. No adverse findings. No adverse findings.
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
APPENDIX 2 INDIVIDUAL CLINICAL OBSERVATIONS
Page 23
EB
i
ec crsr
;
Eh
!
PROTOCOL 132-0113: ACUTE DERMAL TOXICITY STUDY OF 1-7485 APPLIED TO SPRACUE-DAWLEY RATS
A_PP_END_IX_2:_C_LIN_IC_AL _OB_SER_VA_TIO_NS _FOR_S_TUD_Y D_AY_1 _- I_ND_IVID_UA_L _- F_EMA_LE_RA_TS_ _ _ _ _ _ _ _ _ _ _ - - - - - - . - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
.S.T.U.D.Y. .D.A.Y. .1. . . . . . . . .D.O.S.E. .G.R.O.U.P. .1. . . . . . . .2.0.0.0. .M.C./K. C. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
R. A. .T. #. . . . .P.R.E.D.O.S.E. . . . . . . . . . . . . .1. .H.O.U.R. .P.O.S.T.D.O. S.E. . . . . . . . . . . . .2. .H.O.U.R.S. .P.O.S.T.D.O.S.E. . . . . . . . . . . . 3. . H. .O.U.R.S. P. O. .S.TD. O. .S.E. . . . . . . . . . . 4. . .H.O.U.R.S. .P.O.S.T.D.O.S.E. . . . . . . . . . . . . . . . . . . . . . . .
6988 APPEARS NORMAL
APPEARS NORMAL
RED MATERIAL AROUND NOSE RED MATERIAL AROUND NOSE RED MATERIAL AROUND NOSE
6990 APPEARS NORMAL
RED MATERIAL AROUND NOSE RED MATERIAL AROUND NOSE RED MATERIAL ARMJWD NOSE RED MATERlAL ARWND NOSE
6992 APPEARS NORMAL
RED MATERIAL AROUND NOSE RED MATERIAL AROUND NOSE RED MATERIAL ARWND NOSE RED MATERIAL AROUND EYES
RED MATERIAL AROUND NOSE
6996 APPEARS NORMAL
APPEARS NORMAL
APPEARS NORMAL
APPEARS NORMAL
RED MATERIAL AROUND NOSE
6996 APPEARS NORMAL
RED MATERIAL AROUND EYES REO MATERIAL ARWND EYES RED MATERIAL ARWND EYES RED MATERIAL AROUND EYES
_-________________________________R_ED M_ATE_RIA_L A_RWN_D N.OS-E -------------------
9(L1
tD
h,
(3,
W a,
m(D
Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
APPENDIX 3 INDIVIDUAL BODY WEIGHTS
Page 28
m-0
(Q (D
h)
(D
-0 Lu
(D (D
w
0
Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
APPENDIX 4 CERTIFICATE OF ANALYSIS
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
Certificate Of Analysis
CJ79S03-K' (PFBS), Lot 2
March 10,2000
This sample wps analyzed using L W . 'H-NMR, '%Nh.nz. md elemental analyses techniqua. The results
of these testsshow the sample to contain the following weight paunt composition:
CF~(CF&S4K' *
0.04 x
J'-
97.86 %
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
APPENDIX 5 PROTOCOL
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
Primedica Redfield Protocol
Protocol Numiur: 132410
ACUTE DERMAL TOXICITY STUDY OF T-7485 APPLIED TO SPRAGUE-DAWLEY RATS
PROPOSAL NO.: 132-010
OBJECTNE: Determine the acute effect(s) of a single dermal application of T-7485 to Sprague-Dawley rats with a fourteenday recovery.
LOCATION OF STUDY AND CONDITIONS OF TESTING:
It is the intention of 3M CorporateToxicology, through the conduct of this study, to generate animdsafety data that may be submitted to regulatory authorities. Primedica Redfield, 100 East h o n e S-
Redfield,Arkansas, 72132,is accreditedby AAAL4C 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. PrimedicaRedfieldwill follow all requirementsspealied
in this approved protocol and all applicable governmentalregulations regarding G o d Laboratory Practices as well as Prirnedica RedfieM's Standard Operating Procedures. Changes in the pmtoco( may be made by consultation with and approval from 3M CorporateToxicology followed by written verifialion of the change. 3M Corporate Toxicology reserves the right to inspect facilities and procedures used for this study by means of announcedor unannounced site visits. Primedica Redfieldwill nobfy 3M Cocporate 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, MN 55133-3320
SPONSOR'S REPRESENTATIVE:
Ph.D., DABT, CIH Senior Laboratory Manager Telephone: FAX
TESTING FACILTTY:
Primedii 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. PrincipleInvestigator: Karen Tranter. R.A. LATG Veterinarian: Allan Manus, D.V.M., M..%., ACLAM
Pathobgist TED
PROPOSED STUDY DATES:
ExperimentalStart Date: July 27,2000 ExperimentalTermination Date: August 10,2000 Draft Report Date: September 7,2000
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7m00
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
Primedica Redfield Protocol
Protocol Number: 132410
1.
REGULATORY COMPLIANCE AND QuAuTYASSURANCE
This study will be conducted in accordance with the following Good Laboratory Practice RegulationslStandardslGuidelines:
H 21 CFR 58 E) C(81)30(Final) (OECD)
The Quality Assurance Unit, in accordance with Primedica Redfields Standard Operating Procedures (SOPS),will audl the protocol. study conduct, and the final report
2.
INSTI-MWNALANIMALCAREAND USECOMMITTEREMEW
The protocol will be reviewed and approved by Prirnedika Redfields 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 unnecessarily duplicate any previous work
3.
MA~UWLS
TESTMATERIAL
lDEN77FlcAnoN
T-7485 (Potassium perfluorobutane sulfonate, PFBS) The lot number will be listed in the taw data and final repon
F w s i c u D ~ s c R i f n o NWhite powder
VEHlW /DENTIFIcAnON
1% carboxymethylcellulose (aqueous, mediumviscosity)
The lot number will be listed in the raw data and final repon
PHYSI~ADLEscRiPnoN White powder
STORAGECoNOmWs
The bulk vehicle and test materialwill be stored in its original container and will be stored at room temperature.
ANALYTICACLHEMISTRY
The Sponsor assumes responsibility f o r characterization (identity. purity,
and stability) of the bulk test material (including under test conditions). Informationon the composition and method of synthesis of the bulk test material will be held by the Sponsor. The doses, unless otherwise stated, will be calculated assuming the test material to be 100% pure.
DOSE FREPARAT~OANND corvcm-mnm
Doses will be weighed by Primedica Redfield on the day of use. The carboxymethylcellulosewill be prepared at 1%in deionized water and may be prepared prior to Study Day 1.
INVENTORY
The control and test materials will be inventoriedwhen received at the testing laboratory, and a record of all test material use will be maintained.
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
Primedica Redfield Protocol
Pmlocol Number: 132-010
TESTMATERIARLETENTION
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 rnatenalwill be packed in a suitable
container to maintain the conditions specified by the Sponsor during transit plus an adequate margin of satety for any transit delays.
9 F E n PREU\UllONS
General safety precautions as required by Primedica Redfidds policies
and procedures will be followed. The S p w o h Representativewill be
notified of any personnel exposures requiring a physician's examination or care.
4.
ANIMALS
Species: Strain/Source:
Age at Initiation: Number and Sex:
Identification:
Rat Crl:CD.(SD) albino rats, Charles River Labwatories. Inc. Six to eight weeks 15 male and 15 fernale (female nulliparous and non pregnant) Uniquely numbered ear tag and cage card
ANIMALHUSBANDRY
HOUS/ffi
The animals will be individually housed in stainless steel cages. The cages conform to standards set forth in the Guide for the Care and U s e
of Laboratow Animals, National Academy Ress, Washington, D.C., 1996.
FEED
Teklad Certified Rodent Diet My8728 will be prwided ad libitum. 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 ad libitum. Samples of the water are
anatyzed for total dissolved solids, hardness, specified microbiological content. and for environmental contaminants. Results of these analyses are on file at Primedica Redfield.
CONTAMIMNTS
There are no known contaminants in the feed or water that would be expected to interfere with this study.
Environmental controls are set to maintain a temperature of 18' to 26C (64"to 79F) with a relative humidity of 30% to 70%. These parameten are recorded at least once daily. A 12:12 hour lightdark cycle is maintained.
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I
Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
Primedica Redfield Protocol
Protocd Number: 132-010
A C W M TlON
Animals will be acciimated for a minimum of seven days prior to the study
start An adequate number of extra animals will be purchased so 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. During pretest the animals will be habituated to the collars used during the study at least three times increasing the minimum time period to at least six hours.
J u s n m n o N FOR SPECIES SELECTIONAND NUMBERoF ANIMALS
Rats are an animal model for acute toxicity studies of thi type. The
number of animals assigned to this study represents the minimum required to meet the objective@)of the study and the OECD guidelines (M02).
5.
EXPEWVENTAL ME~HOOOLOGY
SNDY DESIGN
I 1 I 1 Group
Group
Dosage Level Number of Animals
Number
Desianation
(maika)
Males I Females
I
1
I Hiah-dase I
2000
I5 I 5 I
A S S I G N K M TO GROUPS
For the first phase (Group l)n,o random specik procedurewill be used. If it is necessary to treat Groups 2 and 3, animals will be randomly assigned to groups by a computerized weightordered dsbibution such that individual body weights will not exceed S20% of the mean weight for each sex. Only nalve animals will be used.
Hair will be removed from the domal area of the trunk of the test animals
by clipping at least one hour prior to test material adminisbation and as needed thereafter. Care will be taken tu avoid abrading the skin, which
could alter its permeability. The dose site and an area of adjacent
untreated skin will be marked at the comers with an i n d d i k marker at
the time of dosing. The location of the test and control sites will be
documented in the raw data.
FREQUENCYANDDURAnON OF ADMlNlSTRAnON
A single dosage level of 2000 mg/kg of test material will be applied to
Group 1 animals on Study Day 1. The test material will be wetted to a
paste with medium viscosity carboxymethylcellulose and applied uniformly over an area that is approximately 10% of the body surface area. Test materialwill be held in contact with the skin with a porous gauze dressing and non-irritating tape throughout a 24-hour period. The test material will be further covered in a suitable manner to ensure that the animals cannot ingest the test material. Elizabethan collars will be applied and worn during the exposure period. After the 24-hour exposure
period the test material will be removed with warm water and gauze, and
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
Primedica Redfield Protocol
Proloco1 Number: 132410
the animals observed at least 14 days. If deaths occur, then a full study will follow and consist of applying 1000and 500mgkg of test material with a 14day recovery. If no deaths occur, the study will be terminated. Individual doses will be calculated using the body weights collected on the day of dosing.
JUSSlFlCATlONFOR ROUTE OF ADMlNlSTRATlONAND DOSE LEVEL
In the assessment and evaluation of the toxic characteristics of a substance, determination of acute dermal toxicity is useful where exposure by the dermal route is likely. Dose levels were chosen by the OECD guideline #402.
ANTEMORTEM OSSERVAnONS
CLINICAL OSSERVATIONS
Observations will be perfbnned and recorded twice daily (a.m. and p.m)
for moribundity and mortality.
Clinical observations will be recorded at least once each day and when a change is noted. Observations will be recorded predose and
approximately hourly for four hours postdoseon the first day of dosing then daily thereafter for at least 14 days. Observations should include changes in the skin and fur. eyes, and mucuus membranes, and also respiratory, circulatory. autonomic and central nervous system, somatomotor activii, and behavior pattern. Partiiular attention should be directed to observation of tremon, convulsions, salivation. diarrhea (liquid feces), lethargy, and coma.
BODY W I G H T S
Body weights will be recorded pretest, on Study Day 1, approximately weekly, and at termination.
POSTMORTEM OBSERVATlOrVS
MORIBUNDANIWLS AND ANlhULS FOUND DEAD
Animals unlikely to survive until the next scheduled observation will be weighed, euthanbed, and necropsied. Animals found dead will be weighed and necropsied. In either case. a complete necropsy will be performedas detailed below. Euthanasiaof moribund animals will'be authorized by the Study Director or Shff Veterinarian with the concurrence of the Study Director.
NECROPSY
All animals surviving to the end of the &dy will be humandy euthanaed via carbon dioxide asphyxiation. Euthanasiawill be performed in accordance with accepted American Veterinary Medical Association ( A M ) guidelines IJ. Amer. Vet. Med.Assoc,. 202:229-249. 19931.
On Study Day 15, the animals (non-fasted) will be sacrificed rotating sequentially through groups for each sex. All animals that die during the study, or are killed at study termination, will be subjected to a complete necropsy examination. A complete necmpsy is defined as examination d the external surface of the body, all orifices, and the cranial, thoracic, and
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
Primedica Redfield Protocol
Protocol Number: 132-010
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.
S r A n s n c A r ANALYSIS
Means and standard deviations will be calculated for all quantitative data.
7. REQoRr
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 final report will be
submitted to the Sponsor approximately two weeks after approval of the draft report. The final report will include all elements required/recommended by the regulations and guidelines.
The report will include, but is not limited to, those items listed below:
Descriptive text of the study objecbve Summary Test material identification (Certificate of Analysis, stability data, and analybcal analysis if any are performed) Methods Results and mndusions Body weights
Body weight changes
Necropsy report A copy of the protocol. all protocol amendments, and all protocol deviations that may affect the integrity of the study
8.
MAINTENANCE OF RAW DATAAND RECORDS
Original data or copies thereof, will be available at Primedka Redfield to facilitate auditing the study during its progress and before acceptance of 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
negotiated with the Sponsor. The Sponsor will be notified prior to disposal of any original study data.
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number. 132-010
Primedica Redfield Protocol
Proiocol Number: 132-010
9.
P U A U ~AVSSURANCERMEW
This is to certify that this protocolhas been reviewed by Quality Assurance.
Quality Assurance Auditor Primedica Redfield
10. APPROVALS
Ph.D., DABT. CIH Senior Laboratory Manager
Date
`study Dirkor Primedica Redfield
~
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Primedica Redfield Protocol Amendment
Protocol Number: 132-010
STUDY NUMBER: AMENDMENT: STUDY TITLE:
DATE ISSUED:
132-010
1
ACUTE DERMAL TOXICITY STUDY OF T-7485 APPLIED TO SPRAGUEDAWLEY RATS
October 20.2000
AMENDMENT # 1
ITEM I : CHANGED FROM: CHANGED TO:
REASON:
Page 6, Section 7, Report
Three copies (one bound, two unbound) of the final report will be submitted to the Sponsor approximately two weeks after approval of the draft report.
Three copies (two bound, one unbound) of the approved final report will be submitted to the Sponsor approximately two weeks after approval of the draft report.
Sponsor request.
APPROVALS:
/"/`y/oc
- .-.
Ph.D., DABT, CIH
Date
Senior Laboratory Manager
3M Corporate Toxicology
Study Director Primedica Redfield
1
Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
APPENDIX 6 PROTOCOL DEVIATIONS
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
PROTOCOL DEVIATIONS There were no protocol deviations that adversely affected the integrity of the study.
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
APPENDIX 7 KEY PERSONNEL
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Acute Dermal Toxicity Study in Sprague-Dawley Rats
Study Number: 132-010
KEY PERSONNEL Study Number: 132-010
Study Director:............................................... Study Coordinator:......................................... Head Technician:........................................... Veterinarian: .................................................. Quality Assurance:......................................... Report Coordination: ..................................... Report Preparation: .......................................
John Seng, Ph.D. Karen Tranter, B.A., LATG Martha Downing, B.S. Allan Manus, D.V.M., M.Sc., ACLAM Val Gartner, B.A. Amy Babb, B.S. Tammy Peckham, Ad. Asst. Tammie Hughey, Ad. Asst.
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