Document pmY68EekgdYZK6z249141ekya

, oe CORNINGHazleton Sponsor: St. Paul,iMinnesota ECEIVE]Y J FINAL REPORT Study Title: DailyDose Oral Toxicity Study with T-6669 in Rats Author: Susan M. Henwood, MS, DABT Study Completion Date: April 30, 1997 Performing Laboratory: Comming Hazleton Inc. Ma3d3i0s1onK,inWsimsacnonBsoiunle5v3a7r0d4 Laboratory Project Identification: CHW 6329-197 Page 10f90 0242%8 ) Lo 2! -_ ocww ee1s97 QUALITY ASSURANCE STATEMENT ---------------------------------------- `This reporthasbeen reviewed bythe Quality Assurance Unit of ComingHazleton Inc.,in accordancewiththeFoodandDrug Administration(FDA)Good LaboratoryPractice Regulations, 21 CFR 58and theOrganisationforEconomic Cooperationand Development (OECD) PrinciopfGloeosd Laboratory Practice, C(81)30(Final). The fo`lmlanoawgienmgeinnts.peWcrtiitotnesnwestraetucsornedpuocrttesdoafnidnsfpiencdtiinognss raenpdorftienddtinogtshaerSetiusdsyuDedirteocCtoorrnainndg Hazleton management according to standard operating procedures. Inspection Dates. From To 12/1796 01/21/97 0228/97 0326/97 0422/97 12/1796 0172197 03/0497 03/3197 04/2397 Phase Protocol Review Necropsy Data Review Report Review Report Rereview Date Reported to Study Director 12/17/96 01/2197 03/04/97 03/31/97 04/23/97 Dateto Management 121796 012197 03/04/97 033197 04/23/97 Shanon, fosbiens { Representative, Quality Assurance Unit -- Hfofer Date 2 c 024d i CHW 6329197 STUDY IDENTIFICATION 5 Daily Dose Oral Toxicity Study with T-6669 in Rats Test Material Sponsor Study Monitor Study Director Study Location Study Timetable Study Initiation Date In-Life Start Date: In-Life End Date Study Completion Date. T-6669 3M Toxicology Services Building 220-2E-02, 3M Center St Paul, Minnesota, 55144-1000 Roger G. Perkins, PhD, DABT 3M Toxicology Services SBtu.ilPaduiln,g M2i2n0n-e2sEo-t0a2,,535M14C4e-n1t0e0r0 (612)733-3222 Susan M. Henwood, MS, DABT Coming Hazleton Inc. P.O. Box 7545 Madison, Wisconsin 53707-7545 (608) 241-7221 Coming Hazleton Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 December 20, 1996 January 2, 1997 January 21,1997 April 30, 1997 3 Coz 428 . cw s9157 Toxicology SusanM. Henwood, MS Diplomate, ABT Study Director KEY PERSONNEL Laboratory Animal Medicine Cindy J. Cary, DVM Diplomate, ACLAM Supervisor `Thomas Ryan Study Toxicologist Sandra L. Haley Study Coordinator Toxicology Operations `Pamela D. Gabris Supervisor `Small Animal Toxicology Dose Formulation Dixie K. Bushee Supervisor Quality Assurance Sherry R. W. Petsel Manager Clinical Pathology Robert L. Hall, DVM, PhD Diplomate, ACVP Clinical Pathologist Ronald Markevitch Supervisor `Anatomical Pathology `ThomasE. Palmer, PhD Anatomical Pathologist Jack Serfort Supervisor Necropsy 4 0242% . CHW 323.197 CONTENTS Page QUALITY ASSURANCE STATEMENT.............cuevueraueannannnnn2s STUDYIDENTIFICATION coven eaeen 3 KEY PERSONNEL...4 OBJECTIVE ....ovoieeicie eee eee 10 REGULATORY COMPLIANCE ove10 MAINTENANCE OFRAW DATAANDRECORDS... 10 TEISdeTnMtiAfiTcaEtRioIn AanLdACNhaDracCtAeRriRzIaEtiR.on.............................o..o.e..i.e..n.i..e.e..i.a..n.e..e.e..n11n00 SRteosreargveeCSoandmiptlieosn.s . ...o ...o i ..s .. .s ... .... ..o1 SaDfIeSPtOySPIrIeOcNaut+ ions .e .....c.i.ie.ie.ie.n.ie.eo.eoeeieieies 1 TenCAON ooo 12 SDToUsDeYLDeEvSelIGSeNlectioonoCH.tera -..-.........oviooiosi ss 12 12 PHloaucseimnegnatnodfMAaniinmtaelnsanicnteo .Groups .....................................1134 DOSEPIEPATAON DOSEABBIYSES +o. eoeos o.oo esses eeeee1e 1 ABnotdeymoWreitgehmtOsbse-rva.tion.s .........v....o.i..v.s..e..s..i.o..e...n....i...i...i....i...s.1155 5 oR42y ; cw 65.197 CONTENTS(Continued) Page "PBRlOoCoEdDUSRamEpSl(eCCoonltlienctuieodn)s ................c.ooureenrsenieinneennI.S Palmitoyl CoAOxidaseActivityAnalysis (Day6).................ceueennn1.6. SStcahteidstuilceadl ASDacIrYiSfEicSe +.1. .... (Day20)........cevvrennainiarinnneiaiiiaeianean11s6]. `BAondtyemWoeritgehmtOsbasnedrvCautmiuolnastainvde SBuordvyivWalei.g_h.tG.a.i.n.s__..._..............................................1177 APRaGlUmOiMtoiyClaCloPAAORxOIiOdRaYseActiv+it-yA-Ra+lySv iS .....e ........r .....ccev unerrnne nnn. 1188) CONCLUSIONS o.oo 18 REFERENCES .......0ouenverenressnsesessnsessnsessnsensesansesanen2s0s PATHOLOGYREPORT...........covoeecesieecnseencennannn2n1 COMMENTS ON THEDATA......ouniiiiiiiiianiieeaininainnanianee2n3a `CODGeEnSe,raAlCBodBesRanEdVADIbIAEVTIASIENOONDSU.N.NS.I.T,.S...............ou.v.i.ui.i.in.n.ii.n.no.iao.n.en.e..n..22..5.4 Codes forClinical Pathology ..................................ooi2u]n AbbreviationsandUnits forClinicalChemistry.................coiueien..2..8 CodesforAnatomicalPathology ..............cevueeeunersneerueneennnns29) TAB1LESummaryofAntemortem OBSErVations ...............................30 2 Summary of Body Weight Data (2)............vvuunuunnennnnnnnnnan3n]. 3 Suom fClinm icalCa hemir stryy Data .......v.nnevnneenneunnnenn3n4. APPENDIX A ........ouiieietisiiaseisiieisiseieneeineisee3en MaterialSafety Data Sheet..........c..ueeerunrenrennanenaaennnae..50 6 co2429 CHW 6329-197 CONTENTS(Continued) Page APPENDIBX .....ooiiiiniteienitiescientenies enna, ST IndividualBody WeightData(8) +... evvevernennennnnninnnions59. IndividualBody WeightGain Data(8) +++ +++ovv.ovevvvennennnonn. 62 APPENDICX .......coovvsieniansssinnsasiescasessessansseneann 66 IndividualClinicalChemistry Data +... vvvvneenioessense6se7n ATTENDED + cosrsrtonsigestosioiot ghtsrsmastantatssareats tus saris 9 IndividualAnimalLiver Weight Values(8) ................................70 IndividualAnimal Pathology Data .........................................70 7 02429 i. ABSTRACT cw e950 . `The purposeof this study wastoassesstheoral toxicity produced when the test material, T-6669, was administered by the oral route (gavage) to male rats for 5 consecutive days. Male Cri:CD(SD)BR VAF/Plus rats were assigned to four groups (10/group). Each group received dose preparations containing the carrier (reverse osmosis water) or 5.0, 14.0, or 42.0 mg of test material/kg of body weightday (mg/kg) for 5 consecutive days at adose volume of mL/kg. Five animals/group were terminated on Da6(tyhe day following the final dose) for hepatic palmitoyl CoA oxidase activity determinations; remaining animals were sacrificed and necropsied on Day 20. Food and water were provided ad libitum. The animals were observed twice daily (a.m. and p.m.) for mortality and moribundity. Additionally, each animal was removed from its cage and observed for clinical signs predose and at approximately 1, 2.5, and 4hours after each dose administration and daily thereafter. Any abnormalorunusual findings were recorded. Body weight data were collected daily on Days 1 through 20. Blood samples were collected on Days -2 (pretest), 6, 9, 15, and 20; serum and cellular fractions were separated and shipped to the Sponsor. On Day 6, five animals/group were sacrificed, and the liver was removed from each animal and weighed. The right lateral lobe of the liver was collected from each animal and weighed, then analyzed for palmitoyl CoA oxidase activity; the remaining liver tissue was also collected and weighed. On Day 20, the remaining five animals/group were sacrificed and subjected to an abbreviated `necropsy; the liver was collected from each animal and weighed. Remaining liver tissues. collected at the Da6y sacrifice and the whole livers collected at the Day 20necropsy `were shipped to the Sponsor. All animals survived to the respective scheduled sacrifice. There were no test `material-related antemortem observations. Mean body weight gains for mid- and high- dose animals (14.0 and 42.0 mg/kg, respectively) were significantly reduced during the dosing period; Day 1 to 6 body weightgainswere 23, 23, 0, and -4 gfor the control, 5.0-, 14.0, and 42.0-mg/kg groups respectively. In general, mid- and high-dose animals showed a recovery in body weight gain over the first 3 days after completion of the dosing period. The overall mean body weight gains (Day 1to 20) were free ofsignificant intergroup differences and were similar for all groups. 8 02430 . caw sszsi97 `Administrationofthetestmaterial at all dose levels was associated with higher hepatic `palmitoyl CoA oxidase activity. Mean palmiCotAooxiydasle activities on Da6y for controlanimalsand animals given 5.0, 14.0, and 42.0 mg/kg were 5, 24, 39,and39 IU/G, respectively. "Therewere no test material-related macroscopic findings. Based on significant increases in the levels of hepatic palmitoyl CoA oxidase activity, an indicationofperoxisome proliferation, at all dose levels tested, the no-effect level of Tat-s66f6o9rSiscloensssetchuatniv5e.0damygs/.kg when administered to male Crl:CD(SD)BR VAF/Plus 9 0243 . cwsss197 OBJECTIVE `Thepurposeofthisstudywasto assesstheoraltoxicityproduced whenthetest material, `T6669, was administered by the oral route (gavage) to male rats for5 consecutive days. REGULATORY COMPLIANCE All aspects of this study were in accordance with the United States Food and Drug Administration Good Laboratory Practice Regulations for Nonclinical Laboratory Studies, 21 CFR 58, and the Organisation for Economic Co-operation and Development PrinciplesofGood Laboratory Practice, C(81)30, with the exception that analysisofthe test material mixtures for concentration, solubility, homogeneity, and stability were not conducted. MAINTENANCE OF RAW DATA AND RECORDS Originalpaperdataand a copy of the final report willbe retained in the archives ofthe `Wisconsin facilityofComing Hazleton Inc. (CHW), for 1 year following the signing of the final report. One year after the signingofthe final report, the Sponsor will determine the final CHW. dispositionofthe materials. Magnetically encoded data will be retained at TEST MATERIAL AND CARRIER Identification and Characterization `Test Material. The test material, T-6669 (FC-143),Lot No. 235, is a white powder and is 93% to 97% ammonium perfluorooctanoate. It was received at CHW on September 25, 1996. Information on synthesis methods, stability, composition, or other characteristics that define the test material is on file with the Sponsor. 10 o243% CHW 6329197 Carrier.Thecarrierwasreverseosmosis waterandwas provbiydCHeWd. Informationonsourceandtreatmentofthereverseosmosis wateris onfilewithCHW. Storage Conditions `Thetest material was stoatrreoodm temperature. Thecarrierwas obtained directly from the in-house water system under ambient conditions. Reserve Samples Reserve samples werenotrequired to be taken. Disposition Remaining test material was retumed to the Sponsor on March 10, 1997. Safety Precautions `Personnel involved with the study wore the apparel recommended bytheCHW Test Material Safety Committee (TMSC) based ontheMaterial Safety Data Sheet (MSDS; Appendix A). CHW Standard Operating Procedures (SOPs) applied for any apparel not covered by the TMSC recommendations. TEST SYSTEM `Test Animal Male Crl:CD*(SD)BR VAF/Plus rats were obtained from the Portage, Michigan, facility of CharlesRiverLaboratories, Inc., on December 5, 1996. The animals were: approximately 8 weeks old and weighed from 303 to 395 g at initiationof treatment (see Protocol Deviations; Appendix A). J`Tuhsetirfaitciastfiroenquently used in safety evaluation studies as a representative of a rodent species. n C 02433 ` CHW 325-197 Identification Each animal was assigned atemporary number upon aival. Before initiation of treatment, an individually coded passive integrated transponder was implanted into each animal. After randomization for placement on test, each animal was assigned a `permanent number, and the transponderwascoded with the permanent number. All data foran animal are recordedunderthese numbers. STUDY DESIGN Animals were assigned to the study accarding to the following design: Group 1 (Control) 2 (Low) 3 (Mid) 4 (High) T6669 Dose Level! (mg/kg) 00 50 140 420 Number of Animals Male 10 10 10 10 a Twahteerc)onotnrloyl.gTrohuepdwosaes vgoivleunmethweacsar5rimerL/(rkegvfeorsrealolsmgorosuipss. b Five animals in each group were sacrificed for hepatic palmitoy] CoA oxidase analysis on Day 6. The remaining animals in each group were observed for reversibility, persistence, or delayed occurrence of toxic effects until sacrifice on Day 20. Dose Level Selection Criteria Doseswere selected based on the resultsofan acute toxicity study. In the acute toxicity study (CHW 61001760), animals were dosed withT-6669at concentrations up to 500 mg/kg. Mortalitywas noted for animals given 500 mg/kg. At 250 mg/kg, clinical signs were limited to red-stained face and wet urogenital area for twooffive females; all 12 c0243/ cw 6328157 animals survived to the scheduled sacrifice. The high dose of 42.0 mg/kg selected for this studyisone sixth ofthe acute toxicity no-adverse-effect level of250 mg/kg. The low-and mid-dose levelswereselectedtoprovidean adequate dose response. PROCEDURES `This study was conducteidn accordance with CHW Protocol TP6785 dated December 20, 1996. The protocol and protocol deviations are in Appendix A. Acclimation Fifty males were received on Decembe5r, 1996, and acclimated in Animal Room 512A for28 days before initiation of treatment. In general, animals in this shipment appeared healthy. During acclimation, the animals were examined for abnormalities indicative of health problems, and body weights were recordedforall animals at randomization. Housing and Maintenance Animal Room S12A was used for ths study. Environmental controls for the animal room were setto maintain 19 10.25C (66 to 77F), arelative humidity of 50% 20%, and a 12-hour light/12-hour dark cycle. The animals were housed individually (except for the first ix days following arrival `when the animals were group-housed) in stainless steel, screen-botiom cages. Certified Rodent Diet #5002 meal (PMI Feeds, Inc.) was providedad libitum. The lot `numbers are recorded in the data. The dieti routinely analyzed by the manufacturer for `nutritional components and environmental contaminants. `Water was provided ad libitum. Samplesofthe water are analyzed for total dissolved solids and specified microbiological content and for selected elements, heavy metals, organophosphates, and chlorinated hydrocarbons. The results are on fle with CHW. There were no known contaminants in the food or waterthatwould have interfered with this study. 13 cozasg aw a9 PlacementofAnimals into Groups "The animals were examined by a laboratoryanimal veterinarian onDecember31, 1996, and found tobe suitableforstudy consideration. On Day -2, animals were ranked by body weight. Animals with body weights exceeding +2 standard deviations of the mean `body weight and those with significant physical abnormalitieswere excluded from selection. The numberofanimals available for selection wasreducedto the required number by excluding animals at the endsofthe body weight range. The animals selected faolrltohcaetesdtu0dyg(r4o0upmsal(e1s0)grwoeurpe)asascicgonreddinag c1o0mtphuetererl-agteinveerraatnekdorfatnhdeormannduommbenrumabnedrs. Group mean body weights were analyzed using Bartlett's test for homogeneityofvariance atthe 5.0% probability level (Winer, 1971) and found to be homogeneous. `Animals not used for the study (10 males) were sacrificed and discarded. Dose Preparation Carrier. The carrier was reverse osmosis water. Test Material. Dose concentrations were based on the test material as supplied. Dose. preparations were mixed daily for dose administration. Each dose level was prepared independently. The specified amount of test material was weighed. A portionofthe carrier was placed into a labeledcontainer and the test material `was added and then mixed with a polytron homogenizer until the test material was. suspended. The mixture was then placed into a calibrated container and the appropriate volumeofcarrierwas addedto achieve the desired concentration. Dose preparations `were mixed for at least 10minutes using a magnetic str plate and stir bar. All completed dose preparations appeared to be solutions. `The dose preparations were kept at room temperature until dosing. Dose Analyses Dose analyses were not done. 1 02436 ames Dose Administration Gavage wasusedbecause historically the oral route has been the rooufchtoiece for `administering a known amountoftest material. `The dose preparations were administered once daily for 5 consecutive days. The dose preparations were givenat adose volumeof 5 mL/kg of body weight. Individual doses `werecalculatedbased on the most recently recordedbodyweights. Animals were dosed at approximately the same time each day. During dose administration, homogeneous test `material solutions were maintained using amagnetic stir plate and stir bar. Antemortem Observations The animals were observedat least twice daily (a.m. and p.m.) for mortality and moribundity. Signs ofpoor health or abnormal behavior were recorded as they were observed. Predoseand Postdose Observations. Eachanimal was removed from its cage and `observed for clinical signs predose and approximately1, 2.5, and 4 hoursaftereach dose. `administration and daily thereafter. Any abnormal or unusual findings were recorded. Body Weights Individual body weight data were recorded on the first day oftreatmeanntd daily thereafter. Blood Sample Collections Blood samples were collected from each animal on Days -2 (pretest; see Protocol Deviations), 9, and 15andat the scheduled sacrifices (Days 6and 20). Animalswere not fasted before blood sampling. Blood samples (approximately 1.5 mL)were collected from a jugular vein of all animals pretest and on Days 9and 15. Atthe scheduled sacrifices (Days 6and 20), as much blasopoo ssid ble was collected from the posterior venacavafrom each animal. At sacrifice, the animals were anesthetized with sodium pentobarbital before blood sample collection. 15 c0243% CHW 6325-197 Followingcollection,the blood samples wereheldat roomtemperature until centrifuged. `Theserumandcellularfractions wereseparatedandstoredin afreezerset to maintain 20C 10 until packed on dry iceand shipped to the Sponsorn January 22, 1997. The Sponsor is responsible for the retention or dispositionofthese samples. Palmitoyl CoA OxidaseActivity Analysis (Day 6) At the Da6y scheduled sacrifice, five nonfasted animals/group were anesthetized with sodium pentobarbital, weighed, and exsanguinated in random order following the blood sample collections. The abdominal cavityofeach animal was opened, and the liver was removed and weighed. The right lateral lobe of the liver was collected from each animal, `weighed, and was flash-frozien liquid nitrogen, then stored in afreezer set to maintain ~70C +10 until analyzed by CHW for palmitoyl CoA oxidase activity. `The remaining liver tissue was collected, weighed, and stored in a freezer set to maintain ~20C 10 unil packed on dry ice and shipped to the Sponsorn January 13, 1997. The Sponsor is responsible for the retention or disposition of these samples. Animal carcasses were discarded without further examination after liver tissue collection. Scheduled Sacrifice (Day 20) On Day 20, the remaining five animals/group were anesthetized with sodium pentobarbital, weighed, exsanguinated, and subjected to an abbreviated gross necropsy in random order following the blood sample collections. The necropsy included a `macroscopic examinationofthe external surfaceofthe body; all orifices; and the cervical, thoracic, and abdominal cavities and viscera. All abnormalities observed were recorded. `The whole liver was collected, weighed, and stored in a freezer set to maintain 20C 10 unil packedondryiceand shipped to the Sponsor on January 27, 1997. The Sponsori responsible for the retentionordisposiotftihoesne livers. No additional tissues were collected, and the animal carcasses were discarded after necropsy. 16 C 0243@ CHW 6329-197 Statistical Analyses Only data collected on or afer the first day of treatment were analyzed statistically. One-wayanalysisof variance [ANOVA (Winer, 1971)] was used to analyze Day I body weights; cumulativebody weight gains (Day 1 10 6 and Day 1 to 20), and palmitoyl CoA oxidase levels. Levene's test (Levene, 1960) was done to test for variance homogeneity. In the case of heterogeneityofvariance at p < 0.05, transformations were used to stabilize the variance. ANOVA was done onthehomogeneousor transformed data.Ifthe ANOVA was significant, Dunnett's multiple comparison t-test (Dunnett, 1964) was used for pairwise `comparisons between treated and control groups. `Group comparisons were evaluated at the 5.0% two-tailed probability level. Group2s through 4 were compared with Group 1 (control). RESULTS Astemortem Observations and Survival Antemortem observations are summarized in Table 1; individualdataare in Appendix B. All animals survived to the respective scheduled sacrifice. There were no test material-related antemortem observations Body Weights and Cumulative Body Weight Gains Body weight data are summarized in Table 2; individual data are in Appendix B. Cumulative body weight gains over the dosing period (Days 1 to 6) and over the study duration (Days 1 10.20) are includedin Table 2. Individual day-to-day body weight gains andindividual cumulative body weight gains for Days 1 to 6 and Days I to20arealsoin Appendix B. Mean body weight gains for mid- and high-doseanimals (14.0 and 42.0 mg/kg, respectively) were significantly reduced during the dosing period; Day 1 10 6 body weight gainswere 23, 23, 0, and -4 g for the control, 5.0-, 14.0-, and 42.0-mg/kg groups 1" 02439 CHW 6329197 respectively. Ingeneral, the mid- and high-doseanimals showed arecoveryinbody `weightgainoverthefirst 3daysaftercompletoiftohne dosing period. The overall body `weightgains(Day 1 to 20) were freeofsignificantintergroupdifferences and were similar for all groups. `Palmitoyl CoA Oxidase Activity Analysis Day 6 palmitoyl CoA oxidase activity data are summarized in Table 3; individual data are in Appendix C. The Pathology Report contains a discussion of the data. On Day 6, palmitoyl CoA oxidase activity was significantly increased for treated animals atall dose levels. Mean hepatic palmitoyl CoA oxidase activitesforthe groups given 00,50, 14.0, and 42.0 mg/kg were 5, 24, 39, and 39 IU/G, respectively. Anatomical Pathology ResultsoftheDay20necropsiesand the Day 6andDay20liver weightdata are in Appendix D. The Pathology Report containsadiscussionofthe necropsy findings. Atnecropsyon Day 20, one animal from the group given 5.0 mg/kg had multiple, dark red foci of variable size intheright lobe of the thymus, and the pelvisofthe left Kidney in one animal from the group given 42.0 mg/kg was enlarged. These were considered incidental findings and unrelated to the test material. There were no macroscopic findings in the remaining animals examined. CONCLUSIONS Based on significant increases in the levels of hepatic palmitoyl CoA oxidase activity, an indication of peroxisome proliferation, at all dose levels tested, the no-effect level of T6669 is less than 5.0 mg/kg when administered to male Crl:CD(SD)BR VAF/Plus rats for S consecutive days. 18 coRa40 . awe SIGNATURES Sandra L. Haley Study Coordinator Toxicology Corning Hazleton Inc. Alls DSiupslaonmMat.e,HeAnBwoTod, MS STotxuidcyoDliorgeyctor Coming Hazleton Inc. / 5-97 Date --4lwlfE Date 19 coz14% REFERENCES CHW 6325197 Dunnett, C. W., "New Tables for Multiple Comparisons with a Control," `Biometrics, 20482491 (1964). Levene, H., "Robust Tests for Equality of Variances," ContritobPurobtabiiloitynansd `Statistics, (eds.) L Olkin tal, Ch. 25, pp. 278-292, Stanford University Press: Stanford, California (1960). `Winer, B. J., "Design and Analysis of Single-Factor Experiments,"StatisticalPrinciples in ExperimentalDesign, Second Ed., Ch. 3, pp. 149-260, McGraw-Hill: New York, New York (1971). 20 02448, PATHOLOGY REPORT ciwensig SUMMARY "The purpose of this study was to assess the oral toxicity produced when the test material, "T-6669, was administered by the oral route (gavage)torats for5 consecutive days. The test material was administeraetd dose levels of 0.0, 5.0, 14.0, and 42.0 mg/kgofbody weight/day (mg/kg). `Administrationofthe test material at all dose levelswas associated with higher hepatic `palmitoyl CoA oxidase activity. Mean palmitoyl CoA oxidase activities on Day 6 for control animals and animals given 5.0, 14.0, and 42.0 mg/kg were 5, 24, 39, and 39 IU/G, respectively. There were no test material-related macroscopic findings. METHODS Four groups of male Crl:CD(SD)BR VAF/Plus rats (10/group) were administered the test materialbyoral gavage fo5r consecutive days at a dose level of 0.0 (control group; receivedreverse osmosis water), 5.0, 14.0,or 42.0 mg/kg. Five animals from each group were sacrificed on Day 6 for hepatic palmitoyl CoA oxidase analyses. The whole liver `was removed and weighed, and the right lateral lobeofthe liver and the remaining liver `were weighed. The analyses were performed on the right lateral lobe of liver, and the remaining liver was frozen for shipment to the Sponsor. All remaining animals were sacrificed and subjected to an abbreviated gross necropsy examination on Day 20. At `necropsy, macroscopic observations were recorded, and the entire liver from each animal `was weighed and frozen for shipmteotnhte Sponsor. Statistically significant differences cited in the Results and Discussionsectionare based on comparisons betweenthe controlandtreated groups. 21 cozaas . _-- RESULTS AND DISCUSSION oCvHWe63n29197 Mortality All animals survived to the respective scheduled sacrifice. Clinical Pathology Individual values for hepatic palmitoyl CoA oxidase activity are in Appendix C. Mean `values and the resultsofstatistical comparisons are in Table 3. OnDay 6, palmitoyl CoA oxidase activity was significantly increasedfor treated animals atall dose levels. Mean palmitoyl CoA oxidase activities for the groups given 0.0, 5.0, 14.0,and 42.0 mg/kg were 5, 24, 39, and 39 IU/G, respectively. Anatomical Pathology Individual anatomical pathology data for the animals necropsied on Day 20 are in Appendix D. Individual liver weight valuesfor the animals sacrificed on Da6y are also in Appendix D. Day 20. At necropsy, one animal from the group given 5.0 mg/kg had multiple,dark red foci of variable size in the right lobe ofthe thymus, and the pelvis ofthe leftkidneyin `one animal from the group given 42.0mg/kg was enlarged. Thesewere considered incidentalfindingsand unrelated to the test material. There were nomacroscopic findings in the remaining animals examined. Pathologists: ftd=o pH Robert L. Hall, DVM, PhD D(CilpilnoimcaalteP,atAhColVoPgy) _#2P77 Date +7 E. Palmer, z Pathologist Date 22 cozaal ,- 00000 ccwweesmsiwsr COMMENTS ON THE DATA Various modelsof calculators, computers, and computer programs were used to analyze data in this study. Because different models round off or truncate numbersdifferently, values in some tables (e.g., means,standarddeviations,or individual values) may differ slightly from those in other tables, from individually calculated data, or from statistical analysis data. Neither the integrity nor the interpretation of the data was affected by these differences. `"Tmhgekugn/idtasfyo.r"dose levels on the Hazleton Path/Tox System (HPTS) `summary tables are `The number of animals listed in the headingofthe summary tables for antemortem observations reflects the number ofanimals assigned to each groupatthe stoafrthte study. "The summary tables for antemortem observations indicate the numberofanimals for `which a condition was observed without regardtothe specific nature, severity, reversibility, numberof incidences per animal, or the length of time the condition persisted. Only observationsforeach animal other than normal are indicated on the `summary and individual antemortem observations tables. HPTS considers the dayofinitiation of treatment as "Day 1, Week 1. Body weight data are enteredatthe startof a study week (e.g., abody weight recorded on Day 1 is considered a Week 1 body weight, abody weight recorded on Day 8 is considered a `Week 2 body weight). Daily body weight gaindataare calculated from day to day and are indicated in the appendix by the end point day (e.g., body weight gain from Day 1 t0 2 isshownas Day 2 gain, Day2 to3 as Day 3 gain, etc.). Cumulative body weight gain data are calculated from the first day of the study to the appropriate day (e.g., Days 1 0 6 andDays 1to 20) andare indicatedin thetablesas 1 - 6 CHNGand 1 - 20CHNG. 23 c0za4 `CODES, ABBREVIATIONS, AND UNITS `General Codes and Abbreviations Codes for Clinical Pathology Abbreviations and Units for Clinical Chemistry Codes for Anatomical Pathology wens Note: The following lists of codes, abbreviations, and units are usedby nCeHeWde.d fSoormteh,isbruetponrott. necessarily all,ofthis information`maybe 24 co2446 CHW 6329-197 General Codes and Abbreviations WK Week. N Number of measurements in a group. Mean; MEAN Asithmetic mean. SD; S.D.; STAND DEV; STANDARD DEV; sd Standard deviation. CHNG Change. PRE/DLY Predose observation; daily observation once dosing is completed. . `Group mean is significantly different from the meanofthe control group (Group 1) at ps005. + Indicatesbodyweidatgathhatwtas analyzed statistically. "NA No value; not applicable; not present. P Present. 25 coza4y cw esis General Codes and Abbreviations (Continued) DISPATCH TBW Observations dispatched from the in-life `moduleofthe Hazleton Path/Tox System (HPTS) to the necropsy module for data collection purposes. Observatioanres duplicatesofthe last in-life observations. `Terminal body weight. #,No. Number. Animal Death Codes: 1 `Terminal sacrifice at Day 6. T `Terminal sacrifice at Day 20. 26 02449 NS QS/QNS NR bl TE RE EE SE CHW 6323-197 Codes for Clinical Pathology GENERAL CODES No sample `Quanity not sufficient N`Toecrhenpiecaitan(jsuamdpglmeenvtoltuomreepneoattstuesftficient forrepeatanalysis) uTneacchcneipctaalbelrerodrat(ai,ns.t&r.u,muennatcocreptteacbhlneiciinasnterrurmoerntthoauttrpeustu,ltssaimnple spilled, entry of invalid data) Recording error (recorded incorrect data, e.g., wrong number, spelling error, incorrect date) Enry error (incorrect keyboard entry) `Sampling error 27 cozaglg cw em Abbreviations and Units for Clinical Chemistry `Test. `Palmitoyl CoA oxidase Abbreviation (Units) PCOAO (IU/G) 28 02450 `Codes for Anatomical Pathology awe Code Definition ANIMAL DEATH CODES 1 `Terminal sacrifice at Day 6 T `Terminal sacrifice at Day 20 "TISSUE ABBREVIATIONS IN GL STOMACH, GL STOMACH, NONGL SALIV GL, MANDIB LN, ANT MES/PANC AUDITORY SEB GL LACRIMAL GLAND, EX HEMATO NEOPLASIA LACRIMAL GL, INT CAVITY, ABDOM SALIV GLLPAROTID LN, TRACHEOBRON Lymph node Gland Glandular stomach Nonglandular Mandibular salivary gland Anterior mesenteric/pancreatic lymph node Auditory sebaceous gland Exorbital lacrimal gland Hematopoietic neoplasia Internal lacrimal gland `Abdominal cavity Parotid salivary gland `Tracheobronchial lymph node 29 cozas} 8 a38 hay mary of vty ton saa (5) a88 1E tptensnT gts cs aH mpsRsistent = - Cod, 4 4 4 . , 8G33o = 2022 3 ERE b A &3 RR IRV. 3 on 23: a * / -_-- APPENDIX A Protocol Deviations Protocol TP6785 Material Safety Data Sheet cwemw 35 02459 Protocol Deviations CHW 6325-197 Protocol. Animals. Weight at InitiationofTreatment. "21010250 g" Actual Procedure. All animals assigned to study exceeded 250 g at initiation: body weights at study initiation ranged from 303 10395 g. Protocol. Blood Sample Collections. Frequency. "Predose (1 day before the initial dose administration to Day 1), on Days9 and 15, atthe scheduled sacrifice intervals (Days 6 and 20), and at unscheduled sacrifice intervals." Actual Procedure. The predose (pretest) blood samples were colleocntDeady -2. `These deviationsarenot expected to have affected the resultsofthe study. 36 COR45! 9 `CORNINGHazleton Sponsor: 3M St. Paul, Minnesota PROTOCOL TP6785 Study Title: 5 Daily Dose Oral Toxicity Study `with T-6669 in Rats Date: December20, 1996 Performing Laboratory: 33`0C1omKiinngsHmazalneBtoounlIenvca.rd Madison, Wisconsin 53704 Laboratory Project Identification: CHW 6329-197 37 C 02469 CHW 6329-197 ! TP6785 Page2 STUDY IDENTIFICATION 5 Daily Dose Oral Toxicity Study `with T-6669 in Rats `Test Material Sponsor SponsorRepresentative Study Director Study Location. Proposed Study Timetable In-Life (Experimental)StartDate In-Life TerminationDate Experimental Termination Date T6669 M `Toxicology Services. `Building220-2E-02, 3M Center `St. Paul, Minnesota, 55144-1000 Roger. Perkins, PhD, DABT Toxicology Services `Building 220-2E-02, 3M Center `St. Paul, Minnesota, 55144-1000 (612) 733-3222 FacsimileNo. (612) 733-1773 `Susan M. Henwood, MS, DABT `Corning Hazleton Inc. P.O.Box 7545 Madison, WI 53707-7545 (608) 241-7221 Facsimile No. (608) 242-2736 `Corning Hazleton Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 January 2,1997 January 21, 1997 To be determined 38 [205/ 1. Study 5DailyDoseOral ToxicityStudywith T-6669inRats CHW 6329-197 TP6785 Page3 2. Purpose Toassess the oral toxicityproducedwhen the test materialis administeredbythe `oral route(gavage)toratsfor 5consecutdiavyes 3. RegulatoryCompliance `This studywillbeconducted inaccordancewith the Food and Drug Administration GoodLaboratoryPracticeRegulationsfor NonclinicalLaboratoryStudies, 2Pr1inCcFipRl5es8aofnGdotohdeOLragbaonriastaotriyoPnrfaoctriEcce,onC(o8m1i)c3C0o,-woiptehrtahteieoxncaenpdtiDoenvtehlaotpamnealnytsisof `the testmaterialmixturesforconcentration, solubility, homogeneity, andstability `willnotbeconducted. 4. QualityAssurance A`sWissucroannscineUfanciitliitnyoacfcCoorrdnanicnegwHiaztlhetthoenSItnacn.da(rCdHOWp)er.atingProcedures (SOPs) of the 5. `Test Material A. Identification T6669 B. PhysicalDescription "Tobe documinethnertawedad ta C. RePspoansiarbnidliiSttyatobfiltyihtey Sponsor(includingundertestconditions). Sampleofs `testmaterial andcarriermixturesfor concentration, homogeneity,solubility, D. StorageConditions Roomtemperature: E. ReserveSamples Reserve samplesoftestmaterialarenotrequiredtobetaken. 39 cozasf CHW 6329-197 TPPa6g7e8dS F. DAinnpiyosuetizontouefsTtmesattMere aitaeld wrialllbe returtnotehde Sponsoraafftteerr completioon n.of G. SAasferetqyuPirreecdbauytiCoHnsWSOPsandpolicies 6. Carrier A. ReHdveenrtsifeiocsstmioosniswater(RO water) B. PChlyesair,ccaollDorelsecsrsilpitqiuoind C. Storage Conditions Ambieat 7. ExperimentalDesign A. Animals (1) SRpaetcies @ Sm CH:CDSD)BR VAF/Plus ) CShoaurrlceesRiverLaboratoriIensc,. YAougnegaadtulltnitofiTaretatimoennt (9 W2e0ig1h0t2at5I0nitiofaTrteaitmoennt ( 4N0ummableersandSex (IdenIntdiifviidcuaatlilyocnodedpassiveintegrated transponderimplants 4 c02469. () Husbandry CHW6329-197 TP6785 Pages () Housing Individual(maybe group-housed during acclimation) Food CertifiedRodent Diet#5002 meal(PMI Feeds, Inc.) ad libitum, `unlessotherwisespecified.`Thefoodis routinelyanalyzedbythe `manufacturerfornutritional componentsandenvironmental `contaminants. (9 Water Adlibitum.Samplesofthewater areanalyzedfortotaldissolved seolleimdesnatsn,dhsepaecviyfmieetdamlsi ,ocrr gaonbociponhotoelnstaaonpndhdcghfaloiortrsc ieenlaseatce,tlded `hydrocarbons. (4) Contaminants `Thereare no knowncontaminantsinthe foodorwaterthatwould intewitrhtfhies srtudey. (9) Eavironment `mE aintn ain v 19ti oco2n5r troCls(o 6f6ortn toh7e7am nFi)ma,e larreoln oamtiwvt ielhlua bmeisdeil ttytoof 50% 220%,and a 12-hourlight/1d2a-rkhcoyculer. ( Acclimation Atle1awesekt: ) Randomization Viacomputergeneratedrandomnumbers forassignmenttogroups. `Theanimalswillbeweighed.Theweightvariationofthe animals selectedforthestudywillnotexceed +2standarddeviationsofthe `meanweights. (10) Justification o`Tfhearraotdiesnftrsepqeuceinetsl.yusedinsafetyevaluationstudiesas arepresentative 41 02463 B. GroupDesignations CHW6325-197 TP6785 Page Grovp 21((CLoonwt)rol) 43((HMiigdh)) DoseLevel* (g/kg) 0500 140 420 Numobf Maelers 1100 10 10 a Tvhecoontwrilolllgbu reoupmmwLi/leklgr.eceive ROwateronly.Thedose b Fp iveaanimalls im noexaici dhagsret oaunpalwoyislilsbyoensaDl caryif6i- .cTedhfC eorreo mainAing paenrismiasltseincnee,aocrhdgerloayuepdwoiclclubrereonbcseeorvfetdofxoircerefvfeercstisbiulnittiyl, `sacrionfDiayc2e0d. C. DosingProcedures (Dosing Route Onlgavage @ HR istoreicfaloa lry,tDohsseionro galRroonuutteehasbee therouteofchoiceforadministering a knownamo ofteustmanterital. @) DFoivseicngDourantiosndaeysocfdouseatdminiistvratieon.Thefirstdayoftreatment waillpbepdesrigonattxheedasisamDmeatyai1m.teeTahecehlddoasyye.swillbeadministered at (4) D`oThseecPornterpoalraantiimoanlswill receive ROwateronly.Testmixtures illbe ipnrepRaOrweadtoenreaatcahsdpaeyciofficadcmoinnciesnttrraattiioonnf.Tohreeatcehsdtomasteelreivaell.wiTlhlbeedomsiexfeodr reeaccohnadneidmbaoldfyorweeaicghhtd.aTyhoefpdroespianrgewditlelsbtemibxatsuerdeosnwitlhlebmeossttorreecdeanttlryoom t`ehmopmeorgaetnueroeuusntteisltamadtermialiminxtuirDeussrwiintlglbdreosmaeaaidtnmtiainiinseotdruanstiion.ng,amagnetic stirplanadsttirbear, Q cozasy ! D. ObservoaftAniimoalns CHW6329-197 TP6785 Page? (1) Clinical Observations `Twicedaily (a.m.andp.m.)formortalitythroughDay20 (a.m.mortality onlyonDay20. Animals will be observedpredose;at approximately1, 2.5,and 4hoursaftereachdose; anddailythereafterforclinicalsigns. Observationsmaybeextendedwhendirected bythestudydirector. @ BodyWeights For randomization,before initiation oftreatment(Day1), daily thereafler,at scheduledsacrifices survivalexceeds 1day). (Days 6and20),and atdeath (when ) BloodSampleCollections (a) Frequency Predose(1 day before the initialdoseadministration to Day1), on. Days 9and 15, atthe scheduledsacrificeintervals(Days 6and20), andatunschsaecridficue il nteervadls () MethodofCollection/oNfuAnmibmealrs Animalswillnot be fasted overnightfor scheduledcollections. j`uBgluoloadrsavemipnleosfa(lalpapnriomxailmsaattetlyh1e.p5rmedLo)sewiinltlebrevcaollalnedcotendDfraoymsa9. and15. A blood sample (as muchaspossible)willalso be collected from the posterior venacavaof cachanimal sacrificed in amoribund condition(ifpossible) andfromeachanimalatscheduled `sac(rDiayfs6iancd 2e0s). `Theblood samples willbe stored at roomtemperature andthen `centrifuged;theseparateserumandcellularfractionswillbe stored in a freezerset to maintain -20C +10. Theserum andcellular fractions willbepackedon dry iceandsent tothe Sponsorwithin 1 `week after in-lifetermination. TheSponsorisresponsibleforthe retention anddispoosftihetsaimpolens. " cozas if CHW 63T2P96-7189S7 Page8 `Theserumandcellularfraction samples willbeshippedto: JaDm . Joe hnss on 3MET.&S Bldg. 2-3B-09 935 BushAvenue `St.Paul,Minnesota 55106 JsamhesDi.opJfothmnhseoesnaomnrplheitss.alternate willbenotifiedregardingthe E. Palmitoyl-CoAOxidaseAnalysis (1) FreaqndNuumbeernofAcnimy als Five animaol nDs ay6/ig n rr ando omourdp er 2) Met ofCh ollo ectd ion Onthe dayof thescheduledsacrifice, nonfasted animalswillbe `Theabdominal cavity of each animal willbeopened,andthe liverwillbe remao ndv weie ghedd. The rightlateral lobeofthe liver willbecollected fromeachanimal,weighed,andflash-frozeninliquidnitrogen. Theliver tissuewillbestored in afreezersettomaintain-70C 10until `Theremainingliverwillbecollected,weighed,andplacedinto afreezer settomaintain-20C 10.TheLiverswillbepackedon dryiceand shippedto JamesD.Johnsonwithin 1 week after in-lifetermination.The `Sponsor is responsible for the retention anddispositionof the livers. Animalswillbediscardedafter scheduledliver tissue collection. Any animalselected forpalmitoyl-CoAoxidaseanalysis that diesor is sacrificedin amoribundconditionwillbesubjected10anabbreviated FOSSnecropsyexamination, andall abnormalitieswillberecorded. `Tissueswillbediscardedafter necropsy. " cozash ( CHW63T2P9e-7189S7 Page F. Termination (anniotmdesaiglnatsedfor palmitooxiydasle-anaClyosiAs) 0) UAwninslyclabhneeisdmuuabljleetdchtaSetadictsrfoifoaiuncnaebsdbdtreneadvdiDoaetraeitdsghsrascorsisfnieccerdiopnsaymeoxraimibnuantdicoonnadintdiaolnl. avnaeesnbtahcneatvoiaz,readnmwdiwaeitxlhlslsaboinedgriuteuicnmioaprteedeaedtsd.o.TbahA rebiwthanoll,ewtei loiibgvheeerm sdaw,cirlbialfbeiecdvecl diolawlitels hcletbpeodes,terior L`i1wvweeiergeshwkeidalf,latbenredippn-lalacickfeeetddeiornmntidonraatyfiirocenee.zaeTnrhsdeeshtStioppmp eaditntoioasiJrnaem-snep2so0nDs s,CiJblo1o eh0fnos,rortnThwheie.thin r`enteecnrtoiposnyo. rdisposofithteliivoerns. The animalswillbediscaradfetedr @ SOcnheDdauyl2e0d,Stahcerainfiimcaelswillbeanesthetizedwithsodiumpentobarbital, 1w0eiagnhaed,bbebdviragtrhoeespsonsvetcerriiooprsvayeenxatacmaivneaa,tiedoxns.aTnghueinaatnedi,emnvdaislullbbjssected n`wehcorloeplsiiveedriwnirlalnbdeocomlolredcetre,da,wnediaglhleadb,noarnmdaplliatcieedsiwnitlolbaefrreecezoerrdseedt.tTo he `tmoaJiantmaeisD-2.0JoChns1o0n%.wiTthheinli1vweersewkialflebreipna-clkfeetdeornmidnraytiicoena.nTdhsehipped `aSnpiomnaslosrwiislrelbspeodnissicbalredfeodratfheerrenteecnrtoiposnyo.rdispositionof thelivers. The G. SSttaattiissttiiccaallaAnnaalylsyessweisllbeperformed onDay ty 1bodyweights,camulative body `wAwiedlielgsbhcetrgc iapitino osn(oDfm asytwai2tp ttihshtrGiocraauolgauhnpr at1ley(rscmeeoinnstaitrsd ioiolns))A.,tatnadcphamlemnitto1.ylG-CorA oox2idtuahsreopluegvhs4els. 8 RAefpionarltreportincluding those ternslistedbelowwil besubmitted. DDeesscroocffitthhrpeectoetnisttirsoyplostanentmdtesitmaoterinals DaPtroecseodufrees xperinitiiatmionaendntertminaatilon T Deasbcoorfufmaionryl tpatloitxa tiycdieafttofaebnci ytdsoo selenvel MaTcabruolsactioopniocfombseearnvabtoidoynsweightsbydoselevel 45 cozasy Pathologyreport Palmitoyl-CoA oxidase levels Statistical findings CHW 6329-197 TP6785 Page 10 9. LOorciagitniaolndoatfa,RoarwcDoapiteas,tRheerceoorfd,sw,ilalnbdeFaivnaiallaRbelepaotrtCHWto facilitateauditingthe studyduringitsprogressandbeforeacceptance of the finalreport. Whthe efia nal reportiscompleted,alloriginalpaperdata,including those itemslistedbelowwill `beretainedin thearchivesof CHWfor a periodof 1 yearfollowingsigningofthe finalreport.Oneyearaftersigningof thefinalreport,alloftheaforementioned `materials willbesenttothe Sponsor,and areturnfeewillbecharged. TheSponsor `may electtohave thematerialsretainedinthe CHWarchives foran additionaltime andCHWwillchargea storagefee. Ifthe Sponsorchoosesto have CHWdisposeof thematerials, a disposal feewill becharged. Protocol andprotocolamendments Dosepreparationrecords In-liferecords Shipping records Bodyweights Randomizationdata Doseadministration `Antemortem observations Anatomicalpathologyrecords Palmiot xidoasy e anlaly-sisCreocorAds Statistical records Samplecollection records Finalreport(orisiggniedncaoply) `Thefollowingsupporting recordswillberetainedat CHWbutwill notbe archived `with thestody data. `Water analysisrecords AnRimeal rfoor matndie fregm ezee p rtee armnper da rhaua tmut irdt eiro teyu corrrerdcosreds Instrument calibrationandmaintenraecnocrdes 46 cozasg PROTOCOL APPROVAL CHW6329-197 TP6785 Page 11 Yb DRiopgleormGat.ePe,rAkiBnTs,PhD SponsorRepresentative 3M ofr let Date Di= plomatHee,aAwBoTod,MS StudyDirector `Toxicology ComingHazleton Inc. dePt Representative Quality AssuranceUnit Coming HazletonInc. Date __J2209c Date a cozabq / CHW6325-197 TP6785 Page 12 ATTACHMEN1T `StatisticalMethods (`LTehevsetaeti,st1i9c6a0lm) ewtihllobdestdhoantewtlo tlebsetfusoervdaarrieadnecsechroimboegdebneeliotwy..LIenvtehnecea'steesotf hveatreiraongceen.eityofvarianceat p <0.05, transformationswil beusedtostabilizethe Antraalnyssfiosromefddvaartiaa.ncIeft[hAeANONVOAV(AWiins seirg,ni1f9i7c1a8n)t],wDiulnlnbeettd'sonteeosntt(Dhuenhnoetmto,ge1n9e6o4)uwsiolrlbe: `usedforpairwisecomparisonsbetweentreatedandcontrol groups. ``bOondey-wweaiygAhNtgOaiVnAsw(iDlalybse2utshreodu(gifhatpeprlmiicnaabtlieo)nt)o,aannadlpyazlmDiatyoyl1b-CoodAywoexiigdhastesl,ecvoemlsu.lative oIfnteh-ewAayNaOnaVlAyssihsoofwcsovsaigrniiafniccean[cAefNoCrOboVdAy(wWeiingehrt,sa1t9i7i16t)i]awtiolnlobfeturseeadtmteonatn(aDlayyze1)b,ody cv`waoerviiagarhnitcasen,cwheioatmdhojtguhesentiemniiettnyitarwlieblmolodbvyeewdseoeinxgethrt(assneaeesoatubhsoehveceto)ev,arnroiogaettnreea.intAsylf.otIrhmfoatuthgiehoA nLsewvieNlnleb'eC stuiessseO tsdfiogbnreVicfaiucAasnet., Teastsquares meanst-test(SAS, 1989)willbe used forpairwise comparisons between. treatedandcontrol groups. `Groupcomparisonswillbeevaluatedatthe5.0% two-tailedprobabilitylevel. DuRnenfeertetn,ceCs. 20:482491 (W1.,964")N.ew Tablesfor Multiple Comperisons with a Control," Biometrics, LSteavteisnteic,sH,,("edRso)bIu.sOtlTeisntesftaolr,ECqhua.l2i5t,ypop.f V2e7r8i-a2n9c2e,sS,"taCnofnotrridbUuntiivoenrsstitoyPrPorbeasbsi:liSttyaanfnodrd, California (1960). ISnsAtSitIuntsetIinnct.e:ICnacr.,yS,ANSo/rtShTCAaTrolUisnear'(1s9G8u9i)d.e, Version6, FourthEd,Vol. 2, p.909, SAS 48 02470 / CHW 6329-197 t TP6785 Page 13 Attachment 1 (Continued) B.J, and. a New York (1971a). is of Si -Factor at ger i ," Statistical Principles ameterSt New York, `Wines, B. J, "AnalysisofCovariance,"SEtatixsticaplPreinsirpleis inmentDaesilgn, Second Ed., Ch. 10, pp. 752-812, McGraw-Hill: New York, New York. (1971b). coza7f DMAATTEARISAHLEETSAFETY n3M Center . -- S5t5.14P4a-u1l0,00Minnesota Ew 6s29.197 (812) 733-1110 eer T=6669 Copyright, ALL rights r1e9s9e8r,veKdi.nesCootpyainMginianngd/oarnddMoawnnluofaadcitnugrinogf Cospany. this inforaation for the purpose of properly utilizing 3M products 1i)s athlelowinefdorpsraotviiodned1tshacto:pied in full With no changes unless 2) dnpiersiiottrrhieabrgurtteheeedmeWcniattphyinstoherobttianhiteneneotdriiogfnironmaolf3eM3,arnrainenasgolda porrofoitthertwhiesreeon. DTIRVAIDSEIONNA:ME:SPECIALTY CHEMICALS DIVISION 10FCN-U1M4B3ERF/LUU.OPR.ACD:. Brand Fluorochesical Surfactant 999888-000222111111-570430089896-770 000000---555111111333555--.010209971463128-.-118 9Z8F9..8-000202101121---600538879181-.-049 0000--5-511113955..u019034o6050-58 SDIOUSCPSUEUMREESDNE:TD:EASu:g1u0sM-ta3y820328,0-,319199966 1. INGREDIENT CAS. NO. PERCENT AAMMMMOONNIIUUM PPEERRFFLLUUGORROOHOECPTTAANNOOTAET.E....1....1.1.1..1.1. 6381253.026.-413.4183.-3g7 AAMMMMOONNIIUUMM PPEERRFFLLUUOORROOHPEEXNATNAONAOTAET.E................1.u.u.,. 6281265195--1417--04 01.1 1 -3 2. PHYSICAL DATA BOILING POINT:.........ceceuen. VAPOR PRESSURE:................ VAPOR DENSITY!......uivivuunns ESVOALPUOBRIALTIITOYNRINATNEA!TEsR.:..u.u.u.u.u.s.u.n.s.s. SPECIFIC GRAVITYiuuvravsennsurs PERCENT VOLATILE:............0. PH N/A N/A N/A Na/ppArec. 0.(4Bul= k)0.5Haters N/A C2, 8 VISCOSITY! u.unrransnaninnnnnss (0.5% Aqueous) N/D MELTING POINTS, 0vvernnnnnnnnn. N/A APPLEiAgRhAtNCcEolAoNrDed0OpOoRn:der; slight odor. Abbreviations: N/D - Not Detersined WA - Not Appiicsbie 50 COR47 AMSuOgsu:stFC23-,143199F6LUORAD Brand Fluorochenical Surfactant CHWP6A3G2E9-1927 "30 FIRE AND EXPLOSION HAZARD BATA TT FFLLAAMSMHABPLOEINTL:I.N.I.T.S....L.E0Lv:e.e.e.n.n0e0s.s. NNo/nA-flasmable AFUUTNORIUGANBILTEIOLNINTIETRSPE-RAUTEULR:E.:............. NNI/AA EXHTaItNeGrU,ISHCIarNbGonHEDdIiAo:xide, Dry chesical, Foam SPWEeCaIArLfuFlIlREprFoItGeHcTtIiNvGePcRlOoCtEhDiUnRgE,S: including helmet, self-contained, apnodsitpiavntes,prebsansdusrearoorunpdresasrausr,e wdaeimsatndabnrdealtehgsi,ngfaacpepamraastku,s,anbdunker coat protective covering for exposed areas of the head. UNSUeSeUALHazFaIrRdEouAsNDDeEcXoPaLpOoSIsOiNtiHoAnZAsReDcSt:ion for products of combustion. 4. REACTIVITY DATA STABILITY: Stable INNCoOtMPAApTpIlBiIcLaIbTlYe- MATERIALS/CONDITIONS TO AVOID: HAZARDOUS POLYMERIZATION: Hazardous polyserization Will not occur, HACZaArRbDOoUnSHDoEnCoGxHdPeOSIaTndIOCNarPbRoOnDUCDTiSo:xide, Oxides of Nitrogen, Hydrogen Fluoride, Ataonia. Ts. ENVIROWRNTAL THFORRATION or SPIObLsLerRvESePOpNrSeEc:autions from other sections. Collect spilled material. Urseesiwdeuet,swePelpaicengincoaspcoluondsedorcwoanttaeirnetro,avoid dusting. Clean up RECIOnMciMnEeNrDaEtDeDIfSnPOaSnALi:ndustrial or commercial Tacility in the presence of Daiscopmobsuasltisbllteernmaattievref:al. DiCsopmobsuestoifonwasptreodpucrtosducwtillin ianclfuadceiliHFt.y Pornitted ta accept cheaical waste. Abbreviations: N/D - Not Determinedst N/A - Not Applicable 9ce2473 HSOS: FC.143 FLUOMD Brand Fluorocheatcal Surfactant August 23, 1996 caw 9.197 PAGE 3 : s. ENVIROMENTAL INFORMATION (continued) Tm ENCVIhReOnNiMcEaNlTAOLxygDeATnA:Dosand (COD) Oxygen Deaand (80020) = Nil; =TheNoirle(t.i0c0a0l70Ogx/ygg)e;n 20-Day Deaand Biochenical (ThoD) = 809./3L2; gW/ga;terFatfhleeaad (MDianpnhoniwa (mPaigmnoap)hal4e3s-hprrEoCpSeOls=) 46906-hmgr/LL;CSGOre=en74A0lgae nG(rgSo/eoLlnenAalsgtareus(cSaeplreincaosrtnrautauac)apr1i4c-odrenyutEuCaS) 4(c-edlalydrEyGS0wei(gchotl)d =so7n3sta)g"ri;sa bioconcentration factor (8CF) for amoniua perfluorooctancate (PFO) = 1.8 REVGUoLlAaTtOiRlYe. oIrNgFaOnRiMcATICOoNs:pounds: Wi. VOC Less #20 & Exoapt Solvents: N/A. bSeifnocree rdeigsuploastailo.ns EPA Hazardous). Uv.aSr.y, EcPoAnsHuazlatrdsopupslicHaabsltee rNeugmublearti=onsNomoer a(uNtohtoruist.ies The components of this product are in cospliance with the cheaicel registration requirements of TSCA, EINECS, COSL, AICS, MITI and KTCCL. EPCRA HAZARD CLASS: FIRE HAZARD: No PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: Yes 6. SUGGESTED FIAST AID EYE CONTACT: Innediately flush eyes with large amounts of water for at least 15 minutes, Get immediate medical attention. SKFIlNusChONTsAkCiTn: with large amounts of water. If irritation persists, get medical attention. INHALATION: sIifgnssi/gsnysn/pstyoanpstoscsonotcicnuure,, craelmlovea ppheyrssiocniatno. fresh air. If IF0S0WAnLoLtOWiEnDd:uce vomiting. Drink two glasses of water. Call a physician, Abbreviations: N/O - NotSerernines N/A - Not Applicable 5 Tee coza7lf MgSDiS:tFC23-,143190F6LUOMD Brand Fluorocheascal Surfactant "7. PRECAUTIONARY INFORMATION uwPeAnGsE1s74 Te EYnE PsROTEcCoTrIOoNe:atact. Hear vented goggles. SK`IANvoPiRdOTsEkCiTnIOcNo:ntact. Wear appropriate gloves khen handling this pmCTaeoetcvreoesrnroininaanalngl.d,epdrc:Aoovteeprca'abtiulritloyson3.f irtguelpbmorbsvoeetrse4cstmianUvdseeeecesTgosnarearosrayontrthfseomorfp{erooeltvlhoeofenwrtitnhtgeshkaminTanotlreocrsvoieneastsl)ai.(cst):ihaormhemeasd BPeolrneetdhsyi3e1neS/Lpooatryvionfyitihedenfsolslnoinoirnigde.mat(Searriasmee;x). "REUvICsFeOenMetMWxiEhilNataDuthEsiDtoanVpEpvNtreToonIptLrimAilaTaaiIttnOeitNoa:nilnociasnlinseostxihoaanudssetmbavetelenostsilvrieeteciooamnpm.pernodpePrdriosveeixedpeasrusorurefpfii1mcesimeatnrty. protection. REANSCvIPooIGnlREtaARaTsOaibRnprYapenratatPshvRiOeTndaEgnCdrToeIfisOnpNsi:iarracbtcooorrrsndeabnmcaaesteeWdriitoahnl.aOiSrHbSAoerlrneeecgtulCaeomtnsietomnoasfr:etthieToun.lflo.lorFlaocseinghighe Cificiency filter respirator, full-face supplied sir rospiremecs PREVENTION OF ACCIOENTAL INGESTION: 0a0reansotthsoatr,ougdhrlinykHoLrhsaSco1kpe WahnednMaUtseirn.g tHhaisshpmraodnudcst.afWtaesrhheexrproisnegd and before eating. RECOMENDED STORAGE! Do not store containers on their sides. Store at room temperature. Keep container dry. Keep container closed when not in use. FIRE AND EXPLOSION AVOIDANCE: **PREVENT MOISTURE CONTAMINATION TO KEEP POWDER FREE FLOWING*,* Keep container tightly closed. No smoking while handling this material. HMIS HAZARD RATINGS: HEALTH: 3 FLAMMABILITY: 0 REACTIVITY: 0 PERSONAL PROTECTION: X (See precautions, section 7.) mcagoreNt EXPOSURE LIMITS VALE WIT AMMONIUM PERFLUOROOCTANOATE.......... /AAAPOYHMNOOTNNIIUUUNMM PPPEEERRRFFLLFUULGORRUOOGPHEERNPOITAAKNNEOORAXTTAEE..N.1O.1.N.1T.11.E.0... 0.01 Woimd 000..114 HnhGoa/mmMss3 Te Aum sane TuA AGGIN TaTHnaA 3am n mvvY Abbreviationsi N/D - Not 53 Determined N/A - Not Applicable 02B 478E " SAuOgSu:stFC23-,14319F9L6UORAD Brand Fluorochesical Surfactant EXPOSURE LINITS (continuad) INGREDIENT VALUE UNIT cu 6325-197 PAGE 5 TYPE AUTH SKIN t*ihneScKlIpuNodtienNgnOtTAimTauIclOoNuc:sontmreLimibbsutrteadineonsuatbnosdtatenhycee,esosvieirtnahdleilrcatabyex!dpoawsiiurtrbheornb'eyY' torhu,endeweurotraeSnKepImNaurnrteiefuensrseertloy, by direct contact with the substance. Vehicles can alter skin absorption. S-JOaAUnCR:GCIEH:OFAonmEeXrPRiOecScaUonRmEmeCnoLdIneMfdIeTreEnxDcApeToAs:uorfeGGouviedrenmleinnteasl Industrial Hygienists "a. HEALTH HAZARD DATA EYEMpaoidCneO,NrTaAttCeeTa:rEiynsg,Irraintdathiaozny: vsiisigonns./sysptoms can include redness, swelling, Asrborne product aay cause eye injury consisting of corneal opacity. SKIPNrodCuOcNtTACiTs: not expected to be rritating to the skin. Msi1lgdnsS/ksiynnptIornrsitactainoninc(laufdteerrepdrnoelsosn,gedsweolrlirnegp,eatanedd contact): itching. May be absorbed through the skin and persist in the body for an extended tive. INHIAlLlAnTeIsOsN:requiring medical by inhalation to moderate attention may quantsties of result from a single this material. exposure May be absorbed by inhalation and persist in the body for an extended time. Single overxposure, above recomnended guidelines, may cause: Irrtnavion (upper respiratory): signs/symptons can include soreness of the nose and throat, coughing and sneezing. Pcaruosleosnged of repeated overexposure, above recommended guidelines, nay Liver Effects: signs/synptons can include yellow skin(jaundice) and tenderness of upper abdomen. Repeated inhalation of airborne product above the exposure guideline can result in elevated organofluoride levels in the blood. Abbreviations: Nib - Not Deterained WIA - hot Appiisabie 4 0247T60T AMSuDgSu:stFC23-,14319F9L6UORAD Brand Fluorochenical Surfactant 5. HEALTH HAZARD DATA (continued) Hw 6329-197 race 6 or 1F_ISnUgAeLsLtOiWoEnD:is not a likely route of exposure to this product. tLhiinsemsatomradyalr.esult fron a single suallowing of a moderate quantity of CANACEmRi:xture of sasoniua perfluorooctancate, asmontua pp(ee3rr8tf2ll5uu-oo2rr6oo-hh1ee)xpatwnaaonsaotafete,ed, ttohsaatamlowbnaiisnuoa 5p3reatrtotslufSoo7rr%oAp2oMnyOteaNarnIscM,atenPoERacFnoLdmUpOaoRmuOsnGodCnTuiAsnNdOuAcTeEd oscia-grynciiefnaiorcgaennstitcuidctyoymptwohauesnrdefowrueenrldeatesindtabttheiensitgisntcuadltyle.ystsiTichguenlriaefriwetcruaesnotrsstc.aorslpInsotuiancdaslerlceyyvnadted T1beiinnbidigitnnugnstuanmhdoarvsepaindron-fhtehudenancloihnveterra,olltshp.anLcaBrpaelsaiesdc,atoinaonntdsh.etesc(tu1ir9sr6e3wnhatonmdKno1cw6ol9se3pdagsrete,uddittehosesaed conducted Jointly by OH and DuPont). MUTNAoGtENmIuCtIaTgYe:nic in Invitro eutagenicity assays. Did transforaation in a masaslian cell transformation ansostayc.ause coll REPNRoOtDUtCeTrIaVtEog/eOnEiVcELOinPHErNaTbAbLitsTOXbIyNSo:ral aceinistration. Fats by gavage or inhalation exposures. Not Teratogentc to OTHAER3HEPArLoTdHuctAATZoAxRiDciItNyFOSRuMnAbTaIrOyN.: Sheet is available. SECTION CHANGE DATES HEADING SECTION CHANGED SINCE ay 20, 1996 TssvE Abbreviations: W/D - Not Determisnsed N/A - Not Applicable or c0247 : ( ASuOgSu:stFO23-,143199F6LUORAD Brand Fluorochenical Surfactant CHW 6329-197 PAGE ITbeHhPeLSIOiETnDTf,eocrtImNaaCtsLiUoDnoIfNiGtn,hethBidUsaTteNMOaTtisesrLuiIeaMdlI.TSEaDf3eMTOt,yMAAKDNEaYStaNISOMhPeWLAeIRtERDA(NNMTASIRDESRS)A,NTiYsEXPobRpeElSiSeEvDe.dOtno KMPhEEeRRtFChOHeRArMNATNtAhCBeEIL3OIRHTYpUOrSRoAdGuEFcItTONFE1SsTSAfGiFEtO.RfAoUrsPeaArRpTaIirCstULirAceRuslpaoPrnUsRiPpbOulSreEposOfeRoraCnOddUeRtSesEruaiOitFnaibnige for ucunasineqr'uaseffleyncetwtihtothdhienoufstehueseanudsoerra'paspplpkilncioacutalitioeondng.eofaGnaid3veconnptrtrohodelu,cvta,ritiestoisymeeofsofsefnawtchitiscoshrswahrteehtor ptahretiucsuelrarevapluuraptoesetahned3Msupirtoadbulectfotro udsoetre'rsaimneethWohdetohferusIet Ss or afpitplifcoarti3on. Di3un1eeprTrrooovritsdh,eesoraeilsnsofstoieronnpasotsisooinrbiaillnitteeyrlaettcihtoartnosniecilnecftothrriaosnaisicnftaorrasmneasrtfvieiorcne,naTy3oMihstastvekocsursentseov.mierresy. riiennfpfororersssaeatntitioaontnioionnbsttahiaesnedtMSoDfSritosnavcaaoisldapabltleaetbaesndesisrsemcaytolrynoatcfcruboremacMay3.. curIrne'natddiatsioinn.a 5 02479 APPENDIX B Individual Antemortem Observations Individual Body Weight Data (g) Individual Body Weight Gain Data (g) cw sus197 57 cozayg 8 a88a@ oS itn wag mi ae 1 a33 "& s GE OE OH OBE OE BOE on on on ou :2222 Fo Aopandix 8 " 38 & LE 3 oF bd 1d & ion oyoicoe uc (1 a82 2 a tit yngee cs gmt 3 i iy Vo3 8 3 i 88a3 - [a ---- og Er ff 3 5 1 4 1 om 5 Por <82g i7g H | : = Ee en es ees ne es ie cn 52 FE 2 2 a3-33 . -_ APPENDIX D ocwweessw97 Individual Animal Liver `Weight Values (g) - Day 6 Sacrifice Individual Animal Pathology Data - Day 20 Sacrifice 69 02498 L FD oEEOED OEE Pm OEE Fr pr, 83 PPogoE oOmE mBmE o8n oonma *3 2 8 @2 22 5 & Individual Animal Pathology oats } 33 & 8oS B2s Ss -33 <3 <S82 EEA Brce Sd GT Se 2 oR TERE 352 8 EERERIA Bo wn, BO) Pee se 8882 <3 -38 2 3 &, a i & orang mariecen tne. Say 40 gastitive : 2 8 {g  emmy PE I aS e @ Gem rr PE 382 g ~ & meee ITI 38aS aS a Ee 52 3 ETE 3. B 8 -- Eotektnt nian aaa ste BR eet LT TT 822 A& en rns ] i o2&