Document e7xnbvYkZROnoOdGJYLyYgKEg

FlazletonInc. box@7545 dison,Wl -53707-7545 4 so@ aliveries3:301 Kinsman Blvd.,Madison, WI 66'0088..24ft14.14471 608.241.7227 Fax S3704 CORNINGHazleton Sponsor: 3M St.Pa4 Minnesota FINAL REPORT StudyTitle: Acute OralToxicitSytudyofT-6669 inRats (OECD Guidelines) Author: StevenM. Glaza Study CompletionDate: January10,1997 PerformingLaboratory: Coming HazletonInc. 3301 Kinsman Boulevard Madison,Wisconsin 53704 LaboratoryProjectIdentirication: CHW 61001760 Pagel of35 13097 COMPLL4,NCE STATEMENT Acute OralTo3dcityStudy ofT-6669 inRats (OECD Guidelines) CHW 61001760 Thisstudywas conducted inaccordancewiththe OrganisationforEconomic Cooperation and Development and Principleosf Good LaboratoryPracticeC,(81)30(Finalw)ith the exceptionthatanalysisofthe testmaterialmixturesforconcentration, homogeneity/solubilitaynd stabiliwtays not conducted. StevenM. Glaza Date Study Director Acute Studies Coming Hazleton Inc. 2 QUALM ASSURANCE STATEMENT CHW 61001760 Thisreporthasbeen reviewedby theQualityAssuranceUnitof Coming HazletonInc.,in accordancewiththeOrganisatiofnorEconomic Cooperationand Development (OECD) PrincipleosfGood LaboratoryPracticeC,(81)30(Final)T.he followinignspectionwsere conductedand findingsreportedtotheStudyDirectorand management. InspectioDnates From To Phase 10/24/96 10/24/96 Necropsy 12/29/96 12/30/96 Data/ReportReview Date Reported to Study Director Date to Management 10/24/96 12/30/96 10/24/96 12/30/96 RepresentativeQ,ualityAssuranceUnit Date 3 STUDY EDENTMCATION AcuteOralToxicitSytudyof T-6669 inRats (OECD Guidelines) CHW 61001760 Test Material Sponsor Sponsor'sRepresentative Study Director Study Location Study Timetable Study InitiatiDoante Experimental(In-lifSet)artDate In-lifEend Date ExperimentalTerminationDate Study CompletionDate T-6669 3M ToxicologyService MedicalDepartment 3M Center,Bldg.220-2E-02 P.O.Box 33220 St.Paul,NIN 55133-3220 Roger G. Perkins,PhD, DABT 3M ToxicologyService MedicalDepartment 3M Center,Bldg.220-2E-02 P.O.Box 33220 St.Paul,MN 55133-3220 (612)733-3222 StevenM. Glaza Coming HazletonInc. P.O. Box 7545 Madison,Wl 53707-7545 (608)241-7292 Corning HazletonInc. '3301 Kinsman Boulevard Madison,WI 53704 October9, 1996 October 10,1996 November 14,1996 January 10, 1997 January10,1997 4 Acute Studies StevenM. Glaza Study Director Manager Steven R. Sorenson Study Coordinator JeffreyB. Mcks In-lifSeupervisor Rose M. Bridge AdministrativeSupervisor Toxicology Support Kathy Myers Manager CalvinL. Horton Supervisor KEY PERSONNEL QualityAssurance SherryR- W. Petsel Manager CHW 61001760 Laboratory Animal Medicine Cindy J.Cary,DVM Diplomate,ACLAM Supervisor Anatomical Pathology Thomas E. Palmer,PhD AnatomicalPathologist Deborah L. Pirkel/ Jack Serfort Supervisors Necropsy Anne Mosher Supervisor PathologyData 5 CONTENTS CHW 61001760 CONPLIANCE STATEMENT ................................................2................ QUALITY ASSURANCE STATEMENT ........................................3............... STUDY IDENTIFICATION ..................................................4................ KEY PERSONNEL ..........................................................5............... OBJECTIVE ................................................................8............... TEST MATERIAL ..........................................................8................ Identificati.o.n............................................................8................ Purityand Stabilit.y.,......................................................8................ Storage and Retention......................................................8................ SafetyPrecautions.........................................................8................ TEST SYSTEM .............................................................9............... Test Animal ..............................................................9................ Housing .......................................................................I.........9................. Animal Diet..............................................................9................ Animal Selectionand Grouping..............................................9................ JustificatifoonrSpeciesSelection............................................1.0............... PROCEDURES ............................................................1.0................ Preparationand Administratioonf TestMaterial................................10................ Reason forRoute ofAdministratio.n.........................................1.0............... Observations.............................................................1.0................ Pathology ...............................................................1.0................ StatisticAanlalyses .............................................................................1...1............... Locationof Raw Data,Records,and FinalReport..............................I..I............... RESULTS ...........................................................................I.I................ Mortality................................................................I.I............... Body Weights ............................................................I.I............... ClinicalSigns............................................................I.I................ Pathology ...............................................................1.2................ DISCUSSION .............................................................1.2................ 6 CHW 61001760 SIGNATLTRE ............................................................12.............. REFERENCE ..............................................................1.3.............. PATHOLOGY REPORT .....................................................1.4.............. TABLE I MortalitySummary .....................................................1.5.............. 2 Individualand Mean Body Weights/Body Weight Gains (g)...................1.6............. 3 IndividuaCllinicaSligns......................................................1..8................. 4 IndividuaPlathologyComments ..........................................2.1............... APPENDIX ...............................................................2.3................ ProtocolTP2069 .........................................................2.4............... ProtocolAmendment No. I ................................................3.3............... ProtocolAmendment No. 2 ................................................3.4............... ProtocolAmendment No. 3 ................................................3.5............... 7 OBJECTIVE CHW 61001760 The objectivOef thisstudywas toassesstheacuteoraltoxicitpyroducedwhen thetest materialisadministerebdy theoralroute(gavage)torats.' Allproceduraltimespresentedinthisreportfallwithintheacceptablerangesas specified intheWisconsinfacilitoyfComing Hazleton(CHW) Inc.StandardOperatingProcedure (SOP). TEST MATERIAL Identification The tesmtateriawlas identifiaesdT-6669anddescribeadsawhitepowder. Purityand Stability The Sponsorassumesresponsibilfiotrpyuritayndstabilidteyterminatio(nisncluding undertestconditions)A.nalysisof thetestmaterialmixtureforconcentration, homogeneity/solubiliatnyd stabilitwyas notconducted Storage and Retention The testmaterialwas storedatroom temperature.Any unused testmaterialwillbe returnedtothesponsorafterissuanceofthefinalreportaccordingto CHW SOP. Safety Precautions The testmaterialhandlingprocedureswere accordingto CHW SOPs and policies. 8 TEST SYSTEM CHW 61001760 Test Animal Young adultalbinoratsoftheCrl:CD (SD)BR straiwnere procuredfrom CharlesRiver LaboratoriesI,nc.on September 23, October 7,October 21, 1996 (Portage,Nfichigan facilitayn)d on October I and October 15,1996 (Kingston,New York facility). Housing Afterreceiptt,heanimalswere acclimatedfora periodof atleast7 days. During acclimationand throughoutthe study,theanimalswere separatedby sex and group housed inscreen-bottomstainlesssteelcages. Environmentalcontrolsforthe animal room were setto maintaina temperatureof 19* to 25*C, a relativheumidityof 50% 20%, and a 12-hourlight/12-houdrark lightingcycle.In caseswhere variations from theseconditionsexistedt,heywere documented and consideredtohave had no adverseeffecton the studyoutcome. Animal Diet The animalswere providedcontinuousaccessto LaboratoryRodent Diet#5001, PNH Feeds, Inc.,and water exceptfor approximately17 to 20 hours beforetestmaterial administratiownhen food,but not water,was withheld.The feedisroutinelaynalyzedby the manufacturerfornutritionaclomponents and envirorunentaclontaminants.Samples of thewater are periodicallaynalyzedby CHW. There were no known contaminantsinthe feed or water atlevelsthatcouldbe expectedtointerferweith or affectthe resultsofthe study. Animal Selectionand Grouping Ten male and 10 femalehealthy,acclimatedrats,weighing from 208 to 269 g and approximately8 to 12 weeks ofage,were assignedtotwo treatmentgroups of250 and 500 mg/kg ofbody weight. Each dose levelconsistedof 5 male and 5 femalerats.The animalswere identifiebdy animalnumber and correspondingeartag throughoutthe study. 9 CHW 61001760 JustificatifoonrSpeciesSelection Historicalrlayt,shavebeenusedasa representatoifvae rodentspecieasndarepreferred by variousregulatoryagencies. PROCEDURES Preparation and Administration of Test Material The testmaterialwas mixed with distillweadterto a concentratioonf 0.025g/mL forthe 250 mg/kg dose leveland 0.050g/mL forthe500 mg/kg dose level.The preparedtest materialmixturesappearedto be solutionsA.n individuadlose oftherespectivteest mixturewas calculatefdoreach animalbased on itsfastedbody weight and administered by gavage ata volume of 10 mL/kg ofbody weight.The testmaterialmixtureswere storedatroom temperatureuntiladministered. Reason forRoute ofAdministration Historicalltyh,eoralroutehasbeen therouteofchoiceforadministerinagknown amount oftestmaterial. Observations Clinicaolbservationsand mortalitcyheckswere conductedat 1,2.5,and 4 hoursaftertest materialadministratioand dailythereaftefror14 days. Mortalitycheckswere conducted twicea day (morningand afternoonf)or13 daysaftertestmateriaaldministratioand againthemorning ofDay 14. Body weightswere determinedbeforetestmaterialadministrati(oDnay 0).Additional body weightswere determinedatDay 7,atterminatioonfthein-lifpehase (Day 14),or at deathwhen survivalexceeded I day. Pathology At terminationoftherespectivien-lifpehaseforeach dose levela,llsurvivinganimals were euthanized.Allanimals,whetherfound dead duringthestudyor euthanizedw,ere 10 CHW 61001760 subjectedto an abbreviatedgrossnecropsyexaminationand any abnormalitiewsere recorded.Afternecropsy,theanimalswere discardedand no tissuewsere saved. StatisticaAlnalyses No statisticaanlalyseswere requiredby theprotocol. Location of Raw Data, Records,and FinalReport The raw data,records,and an originaslignedcopy ofthefinalreportwillbe retainedin thearchivesof CHW inaccordancewithCHW SOP. RESULTS Mortality A summary oftheobservedmortalitiysinTable 1. No mortalitwyas observedatthe 250 mg/kg dose level.Two malesand allfivefemalestreatedat500 mg/kg were found dead within4 daysof testmaterialadministratioNno. othermortalitwyas observed. Based on theobservedmortalityt,heestimatedoralLD5o valueswere determinedto be greaterthan 500 mg/kg formalesand between250 and 500 mg/kg forfemales. Body Weights Individuaalnd mean body weightsand body weightgainsareinTable2. Allsurviving animalsexhibitedbody weightgainthroughoutthestudy. ClinicalSigns IndividuacllinicaslignsareinTable3. Allanimalstreatedat250 mg/kg appearednormal duringthestudywiththeexceptionof two femaleswhich exhibiterded-stainefdaceand/or wet urogenitaalreawithin24 hoursoftestmateriaaldministrationC.linicaslignsof toxicitoybservedintheanimalstreatedat500 mg/kg includedred-stainefdace,yellowstainedor wet urogenitaalrea,hypoactivithyu,nched posture,staggeredgait,excessive II CHW 61001760 salivationa,nd death. The survivinganimalstreatedat500 mg/kg returnedto a normal appearanceby Day 7. Pathology Individualgross necropsypathologyfindingsareinTable 4. A summary reportby the studypathologistison Page 14. There were no testmaterialrelatedlesionsobserved at necropsy. DISCUSSION The acute oraltoxicityof T-6669 was evaluatedinmale and female ratswhen administeredas a singlegavage dose atlevelsof250 and 500 mg/kg ofbody weight. Based on the observed mortalityt,he estimatedoralLD50 valuesformale ratswas determined tobe greaterthan 500 mg/kg and between 250 and 500 mg/kg forfemales. Adi mortalityoccurredatthe 500 mg/kg dose levelwithin4 days oftestmaterial administrationA.llanimalstreatedat250 mg/kg appearednormal duringthe studywith the exceptionoftwo femaleswhich exhibitedred-stainefdaceand/orwet urogenitalarea within24 hours oftreatment.Clinicaslignsoftoxicitoybserved inthe animalstreatedat 500 mg/kg includedred-stainefdace,yellow-staineodr wet urogenitalarea,hypoactivity, hunched posture,staggeredgait,and excessivesalivationA.nimals survivingto theend of the observationperiodexhibitedbody weightgainduringthe study. The grossnecropsy examinationsdid not revealany testmaterialrelatedlesions. SIGNATURE StevenM. Glaza Study Director Acute Studies Date 12 CHW 61001760 REFERENCE 1. "Acute OralTo3dcity,O"rganisationforEconomic Cooperationand Development GuidelinesforTestingof Chemicals,Section4,HealthEffectsN,umber 401,Paris Cedex (February24, 1987). 13 PATHOLOGY REPORT CHW 61001760 There were 10 rats(fivemale,fivefemale)each from dose levelsof 250 and 500 mg/kg necropsied.Allanimalsgiven500 mg/kg, exceptforthreemales,diedon test.The survivinganimalswere euthanizedand necropsiedattheterminationofthe study. The dose level,day of death,and grossobservationsrecordedforeach animal are inthe IndividualPathology Comments thatfollowthisreport. At necropsy,therewere few findingsand allof thesewere consideredincidentaalnd unrelatedto the testmaterial.In theanimalsthatdiedon test,one male was partially cannibalizedo,ne femalehad multiple,dark brown areasofvariablesizeinthe glandular mucosa ofthe stomach, and both horns oftheuterusinanotherfemale were fillewdith clearfluid.The pelvisof both kidneysinone femalegiven250 mg/kg was observedto be large.There were no visiblelesionsintheremaininganimals. 'TTioomamsasE@.EPP.a'l@m@er@-@T,g-D Pathologist (61001760.fin) 121696 4?7 Date 14 Table I MortalitySummary Dose Level (mg/kg) Sex MortalityResults No. Died/No. Dosed* 250 m 015 250 F 0/5 500 m 2/5,Days 3',4' Soo F 5/5,Days 0',1',3',4' Superscripntumber indicatensumber of animalsfound dead on theindicateday. CHW 61001760 15 Table 2 Individualand Mean Body Weights/ Body Weight Gains (g) CHW 61001760 Animal Number Day 0 Weight C 12551 269 C12550 269 C 12611 248 C12612 259 C12613 253 Mean 260 Day 7 Weight Gain* Males (250 mg/kg) 330 61 325 56 307 59 296 37 296 43 311 51 Day 14 Weight Gain* 383 114 339 70 375 127 354 95 375 122 365 106 Females (250 mglkg) C12371 261 C12367 248 C12368 235 C12369 259 C12370 252 294 33 285 37 280 45 303 44 287 35 315 54 294 46 285 50 309 50 288 36 Mean 251 290 39 298 47 Gain from Day 0 body weight. 16 Table 2 (Continued) Individual and Mean Body Weights/ Body Weight Gains (g) CHW 61001760 Animal Number Day 0 Weight C12231 213 C 12227 221 C 12228 208 C 12229 217 C12230 215 Mean 215 Day 7 Weight Gain* Males (500mg/kg) 249 36 (167)4 - (168)3 - 271 54 274 59 265 50 Day 14 Weight Gain*- 302 89 - - - - 320 103 334 119 319 104 Females (500 mg/kg) C 12276 248 C 12277 223 C 12278 224 C 12279 246 C 12280 236 (231)' (175)4 t - (234)' (193)4 - - - - Mean 235 - Gain from Day 0 body weight. Animal was found dead on theday of dosing. No body weight required. Value inparenthesesisa dead body weight and was not consideredin calculatintghe mean. Superscriptnumber indicates the day the animal was found dead. Not applicable. 17 Table3 Ind@nd ClWcal 84pa CHW 61001760 Anunal Number Obsefvabon C12551 Appearednormal C]2550 Appearednomial C12611 Appearednomial C 12612 Appearednormal C 12613 Appearednormal Hour Day 1.0 2.5 4.0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Males(250mgAW V V V / /v v v v v v ,v / V V or of v v v ol Of V -1 .1 V V r or V V .1 V V v C12371 Appearednormal C12367 Appearednomal Red-stwned face C12368 Appearednormal C 12369 Appearednorffud Red-sL%inedface Wet urogcnitaAlre, C12370 Appearednormal V Conditioenxisted. - Conditionnotevident Females(250mg/kg) V f V V e -- -- -- - - - -- - -- -- --- - - -- - - - -- VV 18 Animal Number Observation C12231 Appearednomud Red-stamedface Yellow-gained urogenitaalrea C12227 Appeared normal Red-sWned face Hv.poactivity Hunched posture Staggeredgait Found dead C12228 Appeared normal Red-stainefdace Hypoactivity Yellow-stained urogcmtalarea Found dead C 12229 Appearednormal Red-sLunedface C12230 Appearednormal oe Conditioenxisted. - Conditionotevidenl Table3 (CondnuW) bwmdua ciwcalsigm CHW 61001760 Hour 1.0 2.5 4-.0 --- Day 1 2 3 4 5 6 7 8 9 10 11 12 13 F4- Males(500mgft) - - - - - - -I v - - -- - - - -- - - - - - -- - - -- -- - - - - -- e 4e e 19 Table3 (Continued) bidividuaClMdeal Sigm CHW 61001760 Anunal Number Observoion Hour 1.0 2.5 4.0- 1 2 3 4 5 6 7D&yg C12276 Appearednormal Red-stamedfa= Wet uroger@taalm- Hypoactivity Hunched posmm Found dead C12277 Appearednormal Red-stainefdace Wet urogenita-lreHvpoactivity Hunched posture Found dead C 12278 Appearednomua Excessivesalivation v Found dead C 12279 Appearednormal Red-stainefdace Wet urogenitaalrea HypoactMty Staggeredgait Found dead(after clinicaolbservation) C12280 Appearednonmi v Red-stainefdace Wet urogenitaalrea Hypoactivity Hunched posture Staggeredgait Found dead Conditionexisted. Conditionotevidenl Fenutle(s5W mg/W 9 10 11 12 13 14 20 Table 4 IndividualPathology Comments Dose Level: 250 mg/kg of Body Weight CHW 61001760 Animal Number Sex C12551 m C 12550 m C12611 m C 12612 m C 12613 m C12371 F C12367 F C12368 F C12369 F C 123 70 F Not applicable. Test Day Died Sacrificed - 14 Necropsy Observation No visiblelesions. - 14 No visiblelesions. - 14 No visiblelesions. - 14 No visiblelesions. - 14 No visiblelesions. - 14 Both of the kidneys have a large pelvis. - 14 No visiblelesions. - 14 No visiblelesions. - 14 No visiblelesions. - 14 No visiblelesions. 21 Table 4 (Continued) IndividualPathology Comments Dose Level: 500 mg/kg ofBody Weight CHW 61001760 Animal Number Sex C12231 m C 12227 m C 12228 m C 12229 m C12230 m C 12276 F C 12277 F C 12278 F C 12279 F C 12280 F Not applicable. TestDay Died Sacrificed - 14 Necropsy Observation No visiblleesions. 4 - The rightflankwas cannibalized. 3 - No visiblleesions. - 14 No visiblleesions. - 14 No visiblleesions. 3 - The glandularmucosa ofthe stomach hasmultipledark brown areas,up to 2 x I mm. 4 - No visiblleesions. 0 - No visiblleesions. I - The lumen inthebilaterahloms of theuterusisfillewdith clearfluid. 4 - No visiblleesions. 22 APPENDIX ProtocolTP2069 ProtocolAmendment No. I ProtocolAmendment No. 2 ProtocolAmendment No. 3 CHW 61001760 23 CHW 61001760 MEL)LCPL 2E 02 we SampleisubmifteFlorm Tht ibm isioOf &"d ~ suev--m@psw" MRMU&M Bmd* I&SIlng@,FPWW IOSM9 ndkftCM be *at* aff&VsdW conUWI"g ft AcWf &UdiftDopwbnwg afMM) 241-7nl CHWUU*W &via" wM =vrO" aid send to eomkv kazi&Wnkc. 3301 Mnwmn Boulevard Mwbon. VVWoor%Wn63704 SubmM*d by,.!?Qr. r-Q G e@ I Coffpanr. I m FullOLP ConvMnce: 9"'yo4 FDA (21CFFK 58) -w -epArrsc^---4oCFR?92) SanvioNam#: T-(.L(,,q Date Swnple Som: (PAR* Orto,-,1b0/9 Number ofRepoft Requked: 3 -EPA(FIFRA-400FRIGO) -.>tOECD -MAFF Lio@w PhysicalDo&croWn: W" %TC- sft.4-1o SpftlalHancllftProcaWlom: SF-F- MSC>S 10-@7C'07.- Testmaterialpumy and stabuitiyntormilon(kckiftg underis$,condhims) on II*w IM Spo nsor Y*g - w TegtmWtumamtysisiorconcentrslbrv?wrnoggnggy/gtmbiMylobecorldvAgd:_yes. tysponw -bycw &wMIa Disposal:-@@ ReturntOSPOMM At1000WInOgddMW x No ?ickr-ir - S -o rn CF-jiTrA #v c, q,4- owpoee ol acootdingI*ct-W sops Saqwio Storage Requirements: -A- ROOM %"Oratwo - Rdftomtod - oitw At add*ww ow to $P--r (C@W will cormozi Sponsoras tothoseadditionaclherg-). Acute OralTo)"Ity InRate -TP8084 -TP3206 -TP3013 --ILTP2-069 - LIPand down LDSO procedut4i FHSA screen-6.M-SF at 5.0Vkg C@O@ d#M*d sludyN deathoccursat5,0glkg EPA w"n; SM-SF atS.0Og Conduct dofinedaludyP death occursikt5.09^9 OECD @croon;SM-SF *I 5.ogft Conduct dolinodstudyI deathoccursat6.0pq SpecialInstrLcilons; Acut* Dermal TaxicttyinRabbits -TP3207 FHSA *croon:SM.SF of2@ogtkg -TP3016 EPA iocreenS,M-SF at 2.0g&g - Conduct 4*%'wd siudyN deathoccurs&IZO U&g -TP207o OECD =room sm-sf at2.oVkg - CondLom dolkwd study9 deathoccum at2.ogkg Specialinstructions: For CHW Pmioco ssue Data: StutlyDirsoor. Us* Only to-Ct-"%@o Primary &Wn kdwkm -TP3208 -TP3014 -TP2071 -TP4206 -TP7145 PHSA; 6 r4kbbag-*Ibtad*clI lntw sit*lmbbt EPA:6 rabbft-~I swimbbit OECD-.3 rabbfls-ilr4acAtA*Imbbk DOT corrosMW; 6 robbgo.llrftcoliwrabbk PholotoxW, 6 tabbks-2lrftcslkssftabbk (on*sitewithINA emovmo) -SPOCUIkwfucakm: Prknoy Eye Irritation -TP6360 -TP3209 -TP2012 -TP3015 -TP2072 Low-voluffaprocedure;6 t&Wa*sunw"hed FHSA; 8 rabbitsunwashed 1978 EPA; 6 rabbitsunwashed, 3 washed 1992 EPA.*0 mbbho unwashed OECD: 3 rabbitsurmashad 3 (abbftw"hed at4 socords 3 tabboswashed *130 second* SP&CW lnstrudbm: Ouln" PIS Sen*Mzatlon -TP2017 -Tftl64.90 -TP2008 -TP6299 EPA Magnussoiw-Kilpmenroaidmication 01!CD/EC Magnussun.Kffgmennwalmix&Wn SuoW sonskttmlon PhotoaHergeniccontactdormatitb(Artnewong) SpeeW lm!Ar%Xtlmw: While copy-CHW YoNow copy-SubmM*r 24 CHW 61001760 HAZLP.NCC2N W IS C 0 N S IN POST OFFTCE BOX 7S&S bdADISON. WISCONSIN 5370?-?S45 comwm uow"v so@m C-wv Sponsor: 3H St. Paul,Minnesota PROTOCOL TP2069 StudyTitle: Acuteoral ToxicityStudyin Rats (OECD Guidelines) DAU: June 1, 1993 PerformingLaboratory: HazletonWisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 LaboratoryProiectIdentification: HWI 25 CHW 61001760 TP2069 Page 2 STUDY IDENTIFICATION Acute Oral ToxicityStudy in Rats (OECD Guidelines) HWI No. Test Material Sponsor Sponsor's Representative Study Director Study Location Proposed Study Timetable Experimental Start Date Experimental Termination Date Final Report Date C, (Seesample submittalform) 3M ToxicologyServices 220-2E-02 3M Center St. Paul, MN 55144 John L. Butenhoff,PhD 3M ToxicologyServices 220-2E-02 3M Center St. Paul, NN 55144 (612) 733-1962 Steven M. Glaza HazletonWisconsin,Inc. P.O. Box 7545 Madison, Wl 53707-7545 (608) 241-7292 HazletonWisconsin,Inc. BuildingNo. 3 3802 Packers Avenue Madison, Wi 53704 Week of '7 Week of Week of 26 CHW 61001760 TP2069 Page 3 1. Study Acute Oral ToxicityStudy in Rats (OECDGuidelines) 2. Purpose To assessthe acute oral toxicityproducedwhen the test material is administeredby the oral route(gavage)to rats 3. Reaulatory Compliance This studywill be conductedin accordancewith the followingGood LaboratoryPracticeRegulations/Standards/Guidelines: Conduct as a NonregulatedStudy 21 CFR 58 (FDA) 40 CFR 160 (EPA-FIFRA) 40 CFR 792 (EPA-TSCA) C(81)30(Final)(OECD) NotificationNo. 3850, August 10, 1984 (JapaneseMAFF) NotificationNo. 313, March 31, 1982, and as amended by NotificationNo. 870, October 5, 1988 (JapaneseMOHW) All proceduresin this protocolare in compliancewith the Animal WelfareAct Regulations. In the opinionof the Sponsorand study director,the studydoes not unnecessarilyduplicateany previous work. 4. Quality Assurance For regulatedstudies,the protocol,studyconduct,and the final reportwill be auditedby the QualityAssuranceUnit in accordance with HazletonWisconsin (HWI)StandardOperatingProcedures (SOPS) and policies. S. Test Material A. Identification (See sample submittalform) B. PyhnysicaDlescriDtion (See sample submittalform) C. Purityand Stability The Sponsorassumesresponsibilityfor purityand stability determination(sincludingundertestconditions).Samplesof testmaterial/vehicmliexture(s)(ifapplicablef)or concentrations,olubility,homogeneity,and stabilityanalyses will be takenbefore administratioinf requestedby the Sponsor. These samples(if taken)willbe sent to the Sponsor afterexperimentalterminationfor possibleanalysis. 27 CHW 61001760 TP2069 Page 4 D. Storage (See sample submittal form) E. Reservg Samples Studies of less than 4 weeks in experimentalduration will not have reserve samples retained. Reserve sample(s)of each batch/lotof test materialwill be taken if this study is more than 4 weeks in experimental duration. The test material reserve sample will be stored at HWI in a freezer set to maintain a temperatureof below O*C for 10 years per NWI SOP. The Sponsor will be contactedafter 10 years for disposition in accordancewith the appropriateregulatoryGood Laboratory Practices. F. Retention Any unused test materialwill be discarded after issuance of the final report, unless directed otherwiseby the Sponsor. G. Safety Precautions As required by HWI SOPs and policies 6. ExperimentalDesign A. Animals (1) Species Rat (2) Strain/Source Crl:CD-BR/CharlesRiver Laboratories, Inc. Hsd:Sprague Dawley se/Harian Sprague Dawley, Inc. (3) Age at Initiation Young adult (4) Weight at Initiation 200 to 300 9 (5) Number and Sex 5 males and 5 femalesfor the initialdose level 5 males and/or 5 females for any additionaldose levels (ifrequired) (6) Identification Individual numbered ear tag 28 CHW 61001760 TP2069 Page 5 (7) Husbandry (a) Housing Separated by sex and group housed in screen-bottom stainlesssteel cages (heavygauge) (b) LQgd Rodent Choi4* FSOOI (Purina Hills, Inc.) ad libitum except for overnight before test material administration. The food is routinelyanalyzedby the manufacturerfor nutritionalcomponents and environmental contaminants. (c)Water Ad libitum from an automatic system. Samples of the water are analyzed by HWI for total dissolved solids, hardness, and specifiedmicrobiologicalcontent and for selected elements, heavy metals, organophosphates, and chlorinated hydrocarbons. (d) Contaminants There are no known contaminants in the food or water that would interferewith this study. (e) Environment Environmentalcontrols for the animal room will be set to maintain a temperatureof 19 to 25*C, a relative humidity of 50% :t20%,and a 12-hour light/12-hourdark cycle. (f)Acclimation At least 7 days (8) Selection of Test Animals Based on health and body weight according to NWI SOPS. An adequate number of extra animals will be purchased so that no animal in obviously poor health is placed on test. (9) Justificationfor SpeciesSelection Historically,rats have been used as representativeof a rodent species and are preferredby various regulatory agencies. B. Dose Administration (1) Dose Level A single dose of 5,000 mg/kg of body weight will be administeredto five males and five females. If no test material-relatedmortalityis producedat this level, no 29 CHW 61001760 TP2069 Page 6 furthertestingwill be required.If any mortalityoccurs at the 5,000 mg/kgdose level,additionaldose levelsmay be addedat the directionof the studydirectorin order to meet the objectivesof the study. (2)Dose Preparationand Administration All animalswill receivethe sameconcentrationof test mixtureper dose level. If a solid,the testmaterial will be suspendedin an appropriatevehicle. If a liquid, the testmaterialwill be dosedundiluted,usingthe bulk densityto determinethe dose volume. If the materialis an aerosol,it will be dischargedinto a beakerand administereads a liquid. Individualdoseswill be based upon the animal'sbody weight taken just before test materialadministrationa,nd administeredby gavage. The animalswill have food withheldfor 17 to 20 hoursbefore testmaterialadministration.The preparedtest mixtures will be storedat room temperatureuntiladministration. After administrationa,ny remainingtestmixtureswill be discarded. (3)Reason for Route of Administration Historically,the oralroutehas been the route of choice for administeringa known amountof test material. C. Observationof Animals (1)ClinicalObservations At approximatel1y, 2.5, and 4 hoursafter testmaterial administratioannd daily thereafterfor at least14 days for clinicalsignsand twicedaily(a.m.and p.m.)for mortality. Observationsmay be extendedwhen directedby the studydirector. (2)Body Weights Beforeexperimentalinitiation,at 7 and 14 days after testmaterialadministrationa,nd at death (whensurvival exceedsI day) 0. PatholoQy At terminationof the experimentalphase, survivinganimals will be euthanized.All animals,whetherdyingduringthe study or euthanized,will be subjectedto an abbreviatedgross necropsyexaminationand all abnormalitieswill be recorded. After necropsy,the animalswill be discardedand no tissues will be saved. 30 CHW 61001760 TP2069 Page 7 E. StatisticalAnalyses Other than L050calculations(whenapplicable)no statistical analyses are required. Report A final report includingthose itemslistedbelow will be submitted. Descriptionof the test material Descriptionof the test system Procedures Bates of experimentalinitiationand termination Tabulationof mortality data by sex and dose level Descriptionof any toxic effects Tabulation of mean body weights by sex and dose level LD calculations for each sex with 95% confidence intervals rwhen applicable) Gross pathology findings/grosspathologyreport (when applicable) a. Location of Raw-Data, Records. and Final Report Originaldata, or copies thereof,will be availableat NWI to facilitateauditingthe study during its progressand before acceptanceof the final report. When the final report is completed, all originalpaper data, includingthose itemslisted below will be retained in the archives of HWI accordingto HWI SOP. Protocol and protocol amendments DosIe preparation records In- ife records Body weights Dose administration Observations Anatomical pathology records Study correspondence Final report (originalsignedcopy) The followingsupportingrecordswill be retainedat NWI but will not be archivedwith the study data. Animal receipvacclimation records Water analysis records Animal room temperature and humidity records Refrigerator and freezer temperaturerecords Instrument calibration and maintenance records 31 PROTOCOL APPROVAL Joh6 L. lButenhf,f.",PhD@@@ Sponsor's Representative 3M steven . blaz& Study Director Acute Toxicology Hazleton Wisconsin, Inc. Representative Quality Assurance Unit Hazleton Wisconsin. Inc. (TP2069.3M) Date Date CHW 61001760 TP2069 Page 8 32 CHW 61001760 CHW No. PROTOCOL AMENDMENTS C-,f@z c--@r 7 Amendment No. I Effective (Z)c@r@cw_ Portion of Protocol Being Modified: At)cilicabslectionsof the protocol. Reason for Modification:To identifythe locationwhlre the study will be conducted and to reflect a company name change from Hazleton Wisconsin, Inc. (HWI) to Corning Hazleton Inc. (CHW). replacewherever applicablethe following chances Modification: Corning Hazleton Inc. (CHW) 3301 Kinsman Boulevard. Madison. Wl 53704 (GZI/01-07-91) Study Director Approval: 33 CHW 61001760 CHW No. PROTOCOL AMENDMENTS Amendment No. Effective Portion of Protocol Being Modified: Page 4. 6. ExperimentalDesign: A. Animals (21 Strain/Source Reason for Modification: To correctly identifythe nomenclatureused for animals received from Charles River Laboratories.Inc. Modification: Replace this section with the followingchange: (2) Strain/Source Crl:CDO(SD)BR/CharlesRiver Laboratories,Inc. Iri Hsd:Sprague Dawley SDO/Harlan Sprague Dawley, Inc. Study Director Approval: %0 (G21/01-07-91) 34 CHW 61001760 CHW No. PROTOCOL AMENDMENTS 61001760 Amendment No. 3 Effective October 9. 1996 Portion of Protocol Being Modified: Page S. 6. Experimental Design: B. Dose Administration;(1) Dose Level. Reason for Modification: The Sponsor recuested that a level of 500 mg/kq be treatedinitially. Modification: Replace 5.000 mg/kq with 500 mg/kq wherever listed in this section. (G21/01-07-91) Study Director Approval: )@l- -.C4-tt-4-t4@ u 35