Document wrKn5agKZKnwObGN8126g2194
roming HazletonInc. P.O.Box 7545 Madison, Wl 53707-7S4S Deliveries:3301 Kinsman Blvd.,Madison, Wl 608.241.4471 608-241.7227 Fax
S3704
CORNINGHazieton
Sponsor:
3M St.PauL Minnesota
FINAL REPORT
Study Title: AcuteOralToNicitSytudyofT-6684 inRats
(OECD Guidelines)
Author: StevenM. Glaza
Study CompletionDate: January31, 1997
PerformingLaboratory: Coming Hazletoninc.
3301 Kinsman Boulevard Madison,Wisconsin 53704
LaboratoryProjectIdentiirication: CHW 61101149
Page I of29
FEB 3 1997
CONTLLANCE
STATEMENT
Acute Oral To@dcityStudy of T-6684 inRats (OECD Guidelines)
CHW 61101149
This studywas conducted in accordance with the OrganisationforEconomic Cooperation and Development and Principleosf Good Laboratory PracticeC,(81)30(Final).
aStevenM. Glm Study Director Acute Studies Corning Hazleton Inc.
Date
2
QUALITY ASSURANCE STATEMENT
CHW 61101149
Thisreporthasbeen reviewedby theQualityAssuranceUnitof Coming HazletonInc.,in accordancewiththeOrganisatiofnorEcomonic Cooperationand Development (OECD) PrincipleosfGood LaboratoryPracticeC,(81)30(Final)T.he followinignspectionwsere conductedand findingsreportedto theStudy Directorand management
InspectionDates
From
To
12/19/96 12/19/96
01/23/97 01/24/97
Phase Necropsy Data/ReportReview
Date Reported to Study Director
12/19/96 01/24/97
Date Reported to Management
12/19/96 01/24/97
RepresentativeQ,ualityAssuranceUnit
-3
Date
3
STUDY IDENTIFICATTON
Acute Oral Toxicity Study of T-.6684 in Rats (OECD Guidelines)
CHW 61101149
Test Material Sponsor
Sponsor's Representative
Study Director
Study Location Study Timetable
Study InitiatioDnate Experimental(In-lifeS)tartDate in-lifEend Date ExperimentalTermination Date Study Completion Date
T-6684
3M Toxicology Service
Medical Department 3M Center,Bldg. 220-2E-02 P.O. Box 33220 St.PauL MN 55133-3220
Roger G. Perkins,PhD 3M Toxicology Service
Medical Department 3M Center,Bldg. 220-2E-02 P.O. Box 33220 St.PauL MN 55133-3220 (612)733-3222
Steven M. Glaza Coming Hazleton Inc. P.O. Box 7545 Madison, WI 53707-7545 (608)241-7292
Corning Hazleton Inc. 3301 Kinsman Boulevard Madison, WI 53704
November 27, 1996 December 5, 1996 December 19, 1996 January 31, 1997 January 31, 1997
4
Acute Studies
StevenM. Glaza Study Director Manager
StevenR- Sorenson Study Coordinator
JeffreyB. Hicks In-hfeSupervisor
Rose M. Bridge AdministrativSeupervisor
ToxicologySupport
Kathy Myers Manager
CalvinL. Horton Supervisor
KEY PERSONNEL
QualityAssurance SherryR- W. Petsel Manager
CHW 61101149
Laboratory Animal Medicine
Cindy J.Cary,DVM Diplomate,ACLAM Supervisor
Anatomical Pathology
Thomas E. Palmer,PhD AnatomicalPathologist
Deborah L. Pirkel/ Jack Serfort Supervisors Necropsy
5
CONTENTS
CHW 61101149
COWLIANCE STATEMENT ...............................................2...........
QUALITY ASSURANCE STATEMENT ......................................3............
STUDY IDENTIFICATION .................................................4............
KEY PERSONNEL ........................................................5............
OBJECTIVE .............................................................8............
TEST MATERIAL .........................................................8........... Identificat.i.o.n.........................................................8........... Purityand Stabili.t.y.....................................................8............ Storageand Retentior.L...................................................8........... SafetyPrecautions.......................................................8............
TEST SYSTEM ...........................................................9........... TestAnimal ............................................................9............ Housing................................................................9............ Animal Diet ............................................................9............ Animal Selectioannd Grouping.............................................9............ Justificatifoonr SpeciesSelection..........................................1.0...........
PROCEDURES .....................................................I...........1..0.............. Preparationand Administratioonf TestMaterial...............................1.0........... Reason forRoute of Administratio.n.......................................1.0............ Observations...........................................................1.0........... Pathology.............................................................1.0........... StatisticAanlalyses......................................................I.I........... LocationofRaw Data,Records,and FinalReport.............................I..I...........
RESULTS ..............................................................1.1............ Mortalit.y............................................................1.1............ Body Weights..........................................................I.I........... ClinicaSligns...................................................I...............I..I............... Pathology.............................................................1.2...........
DISCUSSION ...........................................................1.2...........
6
CHW 61101149
SIGNATLJRE .............................................................1.2.............
REFERENCE .................................................................1.2............
PATHOLOGY REPORT .......................................................1.3............ TABLE
I Mortality Summary .......................................................1.4............ 2 Individuaalnd Mean Body Weights/Body Weight Gains (g)...................15............ 3 IndividualClinicaSligns..................................................1.6............ 4 IndividuaPlathologyComments ............................................1.7............. APPENDIX .............................................................1.8............ ProtocolT?2069 .......................................................1.9............ ProtocolAmendment No. 1 ...............................................28............ ProtocolAmendment No. 2 ..................................................2.9.............
7
OBJFCTIVE
CHW 61101149
The objectivoef thisstudywas to assesstheacuteoraltoxicitpyroduced when thetest materialisadministerebdy theoralroute(gavage)torats.'
AR proceduraltimespresentedinthisreportfalwlithintheacceptablreangesas specified intheWisconsinfacilitoyfComing Hazleton(CHW) Inc.StandardOperatingProcedure (SOP).
TEST MATERL4,L
Identification The testmateriawlas idenfifiaesdT-6684 and describeadsan off-whitleiquid.
Purityand Stabflity The Sponsor assumesresponsibiliftoyrpurityand stabilidteytermination(sincludinugnder testconditions).
Storageand Retention The testmateriawlas storedatroom temperature.Any unused testmateriawlillbe returnedto theSponsorafterissuanceofthefinarleportaccordingto CHW SOP.
SafetyPrecautions The testmaterialhandlingprocedureswere accordingto CHW SOPs and policies.
8
TEST SYSTEM
CHW 61101149
TestAnimal Young adultalbinoratsoftheCrl:CD (SD)BR straiwnere procuredfrom CharlesRiver LaboratoriesI,nc.on November 4, 1996 (PortageM,chigan facilitayn)d November 12, 1996 (RaleighN,orth Carolinafacility).
Housing Afterreceiptt,heqnim;tlwsere accu=ted fora periodofatleast7 days.During acclimatioannd throughoutthestudy,theanfinalwsere separatedby sex and group housed inscreen-bottomstainlessteelcages.Environmentalcontrolsforthe qnimal room were setto maintaina temperatureof 19*to 25*C, a relativheumidityof 50% 20%, and a 12-hourfight/12-houdrarklightincgycle.In caseswhere variations from theseconditionsexistedt,heywere documented and consideredto have had no adverseeffecton thestudyoutcome.
Animal Diet The animalswere providedcontinuousaccesstoLaboratoryRodent Diet#500 1,PNH Feeds,Inc.,and water exceptforapproximately17 to 20 hoursbeforetestmaterial administratiwohnen food,but notwater,was withheld.The feedisroutinelaynalyzedby the'manufacturefrornutritionaclomponents and environmentaclontaminants.Samples of thewater areperiodicalalnyalyzed.There were no known contaminantsinthefeedor water atlevelsthatcouldbe expectedto interferweithor affectheresultosfthe study.
Animal Selectionand Grouping Fivemale and fivefemalehealthya,cclimatedratsw,eighingfrom 255 to 299 g and approximately10 to 13 weeks ofage,were used fora singledoselevelof 5,000 mg/kg of body weight.The animalswere idenfifibeyd animalmunber and correspondingeartag throughoutthestudy.
9
CHW 61101149
JustificatiofnorSpeciesSelection Historicallrya,tshave been used as a representativoef a rodentspeciesand arepreferred by variousregulatoryagencies.
PROCEDURES
Preparation and Administration of Test Material An individuadlose ofthetestmaterialwas calculatefdoreach animalbased on itsfasted body weight and administeredasa singledose by gavage. The testmaterialwas administereadta volume of 5.05 mL/kg ofbody weightbased on an averagebulk density of0.99 g/mL.
Reason forRoute ofAdministration Historicalltyh,eoralroutehasbeen therouteofchoiceforadministerinagknown amount oftestmaterial.
Observations Clinicaolbservationwsere conductedat 1,2.5,and 4 hoursaftertestmaterial administratioand dailythereaftefror14 days. Mortalitycheckswere conductedtwicea day (morningand afternoonf)or13 daysaftertestmaterialadministratiaond againthe morning ofDay 14.
Body weightswere determinedbeforetestmaterialadministrati(oDnay 0),atDay 7, and atterminatioonfthein-Efephase(Day 14).
Pathology At terminatioonf thein-lifpehase,allsurvivin:g;nim:iwlesre euthanized.Allanimals, whether found dead duringthestudyor euthanizedw,ere subjectedto an abbreviated grossnecropsyexaminationand any abnormalitiewsere recorded.Afternecropsy,the animalswere discardedand no tissuewsere saved.
10
StatisticAanlalyses No statisticaanlalyseswere requiredby theprotocol.
CHW 61101149
LocationofRaw Data,Records,and.FinalReport The raw data,records,and an originaslignedcopy ofthefinarleportwillbe retainedin thearchivesof CHW inaccordancewithCHW SOP.
RESULTS
Mortality A summary ofthesurvivarlateisinTable 1. One femalewas found dead on Day 7. No othermortalitwyas observed.The estimatedoralLDso valuesformale and femalerats were determinedto be greaterthan 5,000mg/kg ofbody weight.
Body Weights Individuaalnd mean body weightsand body weightgainsareinTable2. Aftsurviving anomalsexhibitebdody weightgainwith theexceptionof 3 femaleswhich exhibitedbody weightlossesof I to22 g duringthefirswteek of thestudyand one male which exhibited a weight lossof66 g duringthesecondweek.
ClinicalSigns IndividuacllinicaslignsareinTable3. Allitnimalasppearednormal withtheexceptionof theone femalewhich diedduringthestudywhich exhibiterded-stainefdace, wet/yeUow-stainedurogenitaalrea,hunched posture,tremors,hypersensitivittoytouch, and tonicconvulsions.In additiono,ne male was noted tohave a missingfronttoothon Day 11. Thisankna subsequentlayppearedthinon Days 12 through 14. Thisanimal's weightlossnotedabove isprobablydue to themissingtoothand isnot consideredto be testmaterialrelated.
CHW 61101149
Pathology IndividuaglrossnecropsyfindingasreinTable4. A summary reportby thestudy pathologisitson Page 13. There were no testmaterialrelatedlesionsinany of the animals.
DISCUSSION
The acuteoraltoxicitoyf T-6684 was evaluatedinmale and femaleratsata dose levelof 5,000 mg/kg. The estimatedID50 valueformale and femaleratswas determined to be greaterthan5,000mg/kg. The onlymortalitwyas thedeathofone femaleon Day 7. The onlytestmaterialrelatedclinicaslignsoftoxicitwyere inone male which includeda missingupper rightfronttoothand thinappearanceand thefemalewhich died duringthe studyand includedred-stainefdace,wet/yellow-stainuerdogenitaalrea,hunched posture, tremors,hypersensitivittoytouch,and tonicconvulsionsA.llsurvivinagnimalsexhibited body weightgainwiththeexceptionofthreefemaleswhich exhibitebdody weightlosses of I to 22 g duringthefirswteek of thestudya@d one male which exhibiteda weightloss of 66 g duringthesecondweek. The weightlossinthemale animalwas due to a non-test materiarlelatedconditiorlTherewere no testmaterialrelateldesionsobservedat necropsy.
SIGNATURE
StevenNi Ghm
Date
StudyDirector
Acute Studies
REFERENCE
1. "AcuteOralTo)iicityO,r"ganisationfoErconomic Cooperationand Development GuidelinesfoTrestingofChemicals,Section4,HealthEffectsN,umber 401,Paris Cedex (February24, 1987).
12
PATHOLOGY
REPORT
CHW 61101149
There were 10 rats(fivemales, fivefemales) necropsied. One female died on Test Day 7 and the remaining animalswere euthanizedand necropsiedatthe terminationof the study. The dose level day of death,and gross observationsrecorded for each ;inimalare in the IndividualPathology Comments thatfollowthisreport.
At necropsy,the only lesionsobserved were inthe female thatdied on test.The ventral haircoatwas stainedand the glandularportion of the stomach had multiple,dark red, pinpointfoci.These changes were consideredincidentalfindingsand unrelatedto the test material.There were no visiblelesionsin any of the animalsthatsurvivedto study terminatiorl.
Thomas E. Palmer, PhD Pathologist
(61101149.fin) 010797
Date
13
Table 1 MortalitySummary
CHW 61101149
Dose Level
MortalityResults
(mg/kg)
Sex
No. Died/No. Dosed*
5,000
m
5,000
F
0/5 1/5,Day 71
Superscripntumber indicatensumber of animalsfound dead on theindicateday.
14
Table 2
Individualand Mean Body Weights/ - Body Weight Gains (g)
CHW 61101149
An*mal Number
Day 0 Weight
C15368
264
C15363
270
C15374
299
C1536 7 298
C15337
298
Mean 286
Day 7
Weight
n*
Males (5,000mg/kg)
310
46
305
35
352
53
324
26
346
48
327
42
Day 14 Weight Gain*
334
70
339
69
382
83
312
74
280
-18
341
56
C12896
284
C12898
270
C12905
273
C12903
286
C12908
255
Mean 274
Females (5,000 mglkg)
213
-71
270
0
251
-22
285
-1
247
-8
253 - 20
(209@
-
303
33
294
21
301
15
271
16
292
21
Gain from Day 0 body weight. Value inparanthesesisa dead body weight.Superscripntumber indicatetsheday theanimalwas found dead.
15
Table 3 IndividmdCHdcal ftm
CHW 61101149
Aniffma Number Obswafim
C15368 Appearednomal
C15363 Appm-ed nmmal
C15374 Appearednmmd
C15367 Appeared nomml
C15337
Appearednmnw
Misairwupperrisft fimittoolh
lliinappearance
Hour 1.0 2.5 4.0
or te /
ve
1 2 3 4 5 6 7 9 9 10 11 12 13 14
?dales(5,000mg/kg) v v W, or V,
or Of
of
*I V, t@F
We or le
1(
v 11 1/
vr
v Of
V,
tf Vf
V, to, 4( - - - -
- -- - -- - -- - -
FMSW (!@000mvkg)
C12896
Appearednomul
Yeflow4lained uregenhal-M
Red-WAined fam Wd urogeniw Ere&
... ---
- - -- -- -
60,
00,
...
C12998
Tramm
Hypumubiwto touch
Tonicoonvulsiew
Huwlied posture
Found dead(aftw
dh&d obsovatim)
Appeared nmmal
or
-- - - - - . . .. . .
C12905 Appearednmmal
v
Vol
C12903 Appearednmmal
f .1
V, 61 or V,
C12909 Appeared namud
.1
Conditionexisted Conditionnotevident
or 64, or 100, e V,
400' 00, or
16
Table 4 IndividualPathologyComments Dose Level: 5,000mg/kg ofBody Weight
CHW 61101149
Animal
Number
Sex
C15368
m
C15363
m
C15374
m
C15367
m
C15337
m
C12896
F
C12898
F
C12905
F
C12903
F
C12908
F
Not applicable.
TestDay Died Sacrificed
-
14
Necropsy Observation No visiblleesions.
-
14
No visiblleesions.
-
14
No visiblleesions.
-
14
No visiblleesions.
-
14
No visiblleesions.
7
The haircoatoftheentireventral
surfaceisstainedwith dark red
and drymaterial.The mucosal
surfaceof theglandularportionof
thestomach hasmultipled,ark
red,pinpointfoci.
14
No visiblleesions.
14
No visiblleesions.
14
No visiblleesions.
14
No visiblleesions.
17
APPENDIX
Protocol T?2069 ProtocolAmendment No. I .ProtocolAmendment No. 2
CHW 61101149
18
CHW 61101149
Sample SubmittalForm
7bisfonnIstobe used when su&7*hg &&7p/ssformuthe acutelastingS.pecialtestinngee* can be @a* onangoi by confacttntgheAcuteStudiesDepadamW at (608)241-7292.
CHW Study No.
cz)II dfof
Endose withswwks and send to:
Coming HazMm lrc.
3301 Khwm Boulmrd
Madson, WW=wh 53704
Submktodby@ %
Company.
a-r-i/u
Date Swnple Sat: -
NLrnberd ReportsRequired:. :3
FullGLP comprwm: SanVb Name:
--$-.Yn No -
(oc-
FDA (21CFR 58)
EPA (FIFRA-40CFR 160) -
EPA USCA-40 CFR 792) :@g: OECD
-
(a
MAFF L40M
PhysimINscfipWn: )-L6,@A;4L@
SimcialHancghgPmmnbA- 4-..cA'5ZiWi
Testmateriaplurftaynd stabililtnyfomiatio(nkciucfngtindwtestcondfflms)m Me withSponwr
Testmixtureanalysiisorcorm:ontratorA=wgornky&UNNy tobe conducted:
Yee -
Sample Disposal: ReturntoSpamr atfollowinagddress:
Nc,
Yes by Sponw -
- No by CM
f - tk)o-O
3rv,1;c-b eta Y3 - ;,s-02-
5f
-
Diw@ ofa=rdkv toCM SOPE
Sample Storage Requkwnents: & Room temperature
-Rok4amtW Odor
At additionaaWl toRmw (CHW wM wmct Sponsoras tothese&"kmW charges).
Acute OralToxicityInRats
-TPBOU -TP32D6
.-TP3013
XTP2069
Up and dow, LDSO procedure
FHSA saam; SM-SF at5.09&g ConductderwiedstudyN deathoccursat5.0glkg EPA ser"n;SM-SFat5.0gft Conduct derowd studyI deathoccursat5.0g/kg OECD .on: SM.SF w 5.09&9 Conduct definedstudyI deathoccursat5.0
Speciaklmvidbm:
Acute Dermal ToxicityInRabbits
-TP3207 -TP3016
-TP2070
-
FHSiksaeon-,SM-5F at2.0g*g EPA saeon;SM-SF at2.09/kg
ConductdefinedstudyIdeathoccursat2.0gacg OECD screen;5M-SFat2.0og ConductdefinedstudyIdeathoccursat2-0gtkg
Specialinstructions:
For CHW Us* Only ProtocolIssueDate: Study Diredor:
Prhnwy Sidnkrhdlon
-TP3208 FHSA; 6 mbbks-Iabraded.I kftd Wtokabbk TP3014 EPA; 6 mbbb-I kftd WWmbbk I OECD; 3 rg"$-l ktd shfmbbk
-TP4206 DOT caffowvh-6f.rabbks-NIact sftofrawit -TP7145 Phototmickr6.rabbiLs-i2ntacstkewrabbk
(oneshe withINA exposure) Specialinstructions:
PrknaryEye Irritation
-TP6360 Low-vokmw procedure6; rabbitusnwashed
-TP3209 -TP2012 -TP3015
FHSA: 6 r&Wb unwastwd 1978EPA; 6 rabbitusrwashed.3 washed 1982EPA; 6 tabbb unwashed
-XTP2072 OECD; 3 rabbitmswasw - 3 rabbitwsashed at4 seconds - 3 rabbitwsashed at30 seconds
Specialinstructions:
LiusnonPig Sensitization
-TP2017 -TP6164.EC
-TP2006 -TP6289
EPA Magnusson-rjigmanmaximization
OECD/EC Magnussun-Klignanmaximization Buehlersensitization PhowaNoMenic contacdtermatki(sA-trang)
SpecialInstructions:
Whitecopy-CHW
Yellowcopy-Subminer
19
lV3HAZLEMON WP0STI
B
OF
C,0
F CE
N S IN
8 0 X 7 Ss S
MADISON. WISCONSIN 63707.7645
cofwamo
Sponsor: 3M
St. Paul, Minnesota
PROTOCOL TP2069 StudyTitle:
Acute Oral ToxicityStudy in Rats (OECD Guidelines)
June 1, 1993
PerformingLaboratory: HazletonWisconsin,Inc. 3301 Kinsman Boulevard Madison, Wisconsin $3704
LaboratoryProJectIdentification: HWI
CIRW 61101149
C.,W,
20
CHW 61101149
TP2069 Page 2
STUDY IDENTIFICATION
Acute Oral ToxicityStudy in Rats (OECD Guidelines)
HWI No.
Test Material Sponsor
Sponsor'sRepresentative
Study Director
Study Location
Proposed Study Timetable Experimental Start Date ExperimentalTerminationDate Final Report Date
(It@fIL19
(See samplesubmittalform)
3N ToxicologyServices 220-2E-02 3N Center St. Paul,MN 55144
John L. Butenhoff,PhD 3H ToxicologyServices 220-2E-02 3H Center St. Paul, MN 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, Wl 53704
Week of Week of I Week of /-;1'7-ot"7
21
CHW 61101149
TP2069 Page 3
1. AcuteOral Toxicity Study in Rats (OECDGuidelines)
2. Purpose To assessthe acute oral toxicityproducedwhen the test materialis administerebdy the oralroute (gavage)to rats
3. RtgulatoryComoliance This studywill be conductedin accordancewith the followingGood LaboratoryPracticeRegulations/Standards/Guidelines: Conductas a MonregulatedStudy 21 CFR 58 (FDA) 40 CFR 160 (EPA-FIFRA) 40 CFR 792 (EPA-TSCA) C(81)30(Final)(OECD) NotificationNo. 3850, August 10, 1984 (JapaneseKAFF) NotificationNo. 313, March 31, 1982, and as amendedby NotificationNo. 870, October 5, 1988 (JapaneseMONW)
All proceduresin this protocolare ln.compliancweith the Animal WelfareAct Regulations. In the opinionof the Sponsorand study director,the study does not unnecessarilyduplicateany previous work. 4. OualityAssurance For regulatedstudies,the protocol,studyconduct,and the final reportwill be auditedby the QualityAssuranceUnit in accordance with HazletonWisconsin (HWI)StandardOperatingProcedures(SOPS) and policies. 5. Test Material A. Identification
(Seesample submittalform)
B. PhysicalDeicription (Seesample submittalform)
C. Purityand S&ability The Sponsorassumesresponsibilityfor purityand stability determination(sincludingunder test conditions).Samplesof testmaterial/vehicmliexture(s)(if applicablef)or concentrations,olubility,homogeneity,and stabilityanalyses will be takenbefore administrationif requestedby the Sponsor. These samples(if taken)will be sent to the Sponsor afterexperimentalterminationfor possibleanalysis.
22
CHW 61101149
TP2069 Page 4
D. Storage (See sample submittalform)
E. Reserve Sawles Studiesof less than 4 weeks in experimentaldurationwill not have reserve samples retained. Reservesainple(so)f each batch/lotof testmaterialwill be taken if this study is more than 4 weeks in experimental duration. The test materialreservesamplewill be storedat HWI in a freezerset to maintaina temperatureof below O*C for 10 years per HWI SOP. The Sponsorwill be contactedafter 10 years for dispositionin accordancewith the appropriateregulatoryGood LaboratoryPractices.
F. Retention Any unusedtest materialwill be discardedafter issuanceof the finalreport,unlessdirectedotherwiseby the Sponsor.
G. 'SafetyPrecautions As requiredby HWI SOPs and policies
6. ExperimentalDesian A. Animals (1)Species Rat (2)Strain/SourSe PgT Crl:CD'BFVCharlesRiv,2rLaboratories,Inc. [ ] Hsd:SpragueDawley SO /Harlan Sprague Dawley, Inc. (3)Age at Initiation Young adult (4)Weightat Initiation 200 to 300 g (5)Number and Sex 5 males and 5 femalesfor the initialdose level 5 males and/or5 femalesfor any additionaldose levels (ifrequired), (6)1-dentification Individualnumberedear tag
23
CHW 61101149
TP2069 Page 5
(7) Husbandry
(a)Housing Separated by sex and group housed in screen-bottom stainlesssteel cages (heavygauge)
(b)fDA Rodent Chotr 15001 (PurinaHills, Inc.) ad libitum except for overnightbefore test material administration.The food is routinelyanalyzedby the manufacturerfor nutritionalcomponentsand environmentalcontaminants.
(c)Water Ad libitum from an automatic system. Samoles of the water are analyzed by KWI for total dissolvedsolids, hardness,and specifiedmicrobiologicaclontent and for selectedelements,heavymetals, organophosphates,and chlorinatedhydrocarbons.
(d)Contaminants There are no known contaminantsIn the food or water that would interferewiththis study.
(e)Environment Environmentalcontrolsfor the animalroom will be set to maintaina temperatureof 19 to 25*C, a relativehumidityof 50% +20%, and a 12-hour light/12-hourdark cycle.
(f)Acclimation At least7 days
(8)Selectionof Test Animals Based on health and body weight accordingto HWI SOPS. An adequatenumber of extra animalswill be purchasedso that no animal in obviouslypoor healthis placedon test.
(9)Justificationfor SpeciesSelection Historically,rats have been used as representativoef a rodent speciesand are preferredby variousregulatory agencies.
B. Dose Administration (1)Dose Level A single dose of 5,000 mg/kg of body weightwill be administeredto-fivemales and five females. Ifno test material-relatemdortalityis producedat thislevel,no
24
CHW 61101149
TP2069 Page 6
furthertestingwill be required.If anymortalityoccurs at the 5,000mg/kg dose level,additionaldos levelsmay be added at the directionof the studydirect:rin order to meet the objectivesof the study. (2)Dose Preparationand Administration All animalswill receivethe sameconcentratioonf test mixtureper dose level. If a solid,the testmaterial will be suspendedin an appropriatevehicle. If a liquid, the testmaterialwill be dosed undiluted,usingthe bulk densityto determinethe dose volume. If the materialis an aerosol,it will be dischargedinto a beakerand administeredas a liquid. Individualdoseswill be based upon the animal'sbody weight takenjust beforetest materialadministration,and administeredby gavage. The animalswill have food withheldfor 17 to 20 hoursbefore testmaterialadministration.The preparedtestmixtures will be storedat roomtemperatureuntiladministration. Afteradministrationa,ny remainingtestmixtureswill.be discarded.
(3)Reason for Rou-te-ofAdministration Historicallyt,he oral routehas been the routeof choice for administeringa known amountof test material.
C. Observationof Animals
(1)ClinicalObservations At approximately1, 2.5, and 4 hoursaftertest material administratioannd dailythereafterfor at least14 days forclinicalsignsand twicedaily(a.m.andp.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)
D. Pathology 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.
25
CHW 61101149
TP2069 Page 7
E. StatisticaAlnalyses Otherthan LD50calculations(whenapplicable)no statistical analysesare required.
7. Report A finalreportincludingthoseitemslistedbelowwillbe submitted. Descriptionof the test material Descriptionof the test system Procedures Datesof experimentalinitiationand termination Tabulationof mortalitydata by sex and dose level Descriptionof any toxiceffects Tabulationof mean body weightsby sex and dose level LO calculationsfor each sex with 95% confidenceintervals rw@en applicable) Gross pathologyfindings/grosspathologyreport (when applicable)
8. Lmtion of Raw Data. Records.and FinalReport Originaldata,or copiesthereof,will be availableat HWI to facilitateauditingthe studyduring its progressand before acceptanceof the finalreport. When the finalreportis completed, all originalpaperdata, includingthoseitemslistedbelowwill be retainedin the archivesof HWI accordingto HWI SOP. Protocol and protocol amendments Dose preparationrecords In-liferecords Body weights Dose administration Observations Anatomicalpathology records Study correspondence Finalreport(originalsignedcopy)
The followingsupportingrecordswill be retainedat HWI but will not be archivedwith the studydata. Animal receipt/acclimatiornecords Water analysisrecords Animal room temperatureand humidity records Refrigeratorand freezer temperaturerecords Instrumentcalibrationand maintenancerecords
26
CHW 61101149
PROTOCOL APPROVAL
TP2069 Page 8
Johh L. Butinhoff, PhD
Sponsor'sRepresentative 3M
Steven M. Glaza
u
Study Director
Acute Toxicology
'HazletonVisconsin,Inc.
Representative Quality AssuranceUnit HazletonVisconsin,Inc.
(TP2069.3M)
fz Datb
Date
Date
27
CHW 61101149
CHW No.
PROTOCOL AMENDMEM
Glfcz//Afc)
Amendment No. ,Effective
Portionof ProtocolBeingModified: Applicable-sectionosf the protocol,
Reasonfor Modification:To identifythe locationwhere the studywill be conducted and to reflect a company name change from HazletonWisconsin,Inc. (HWI)to
CorningHazletonInc. (CKW). replacewherever aDRlicablethe followingchanges
Modification:
Corning Hazleton Inc. (CHWI
3301 Kinsman Boulevard. Madison. Wl $3704
(G21/01-07-91) Study DirectorApproval:
28
CHW 61101149
PROTOCOL WNDMEM CHW No. (plj%=) flaf
Amendment No.
Effective
a-7.01"c.
Portionof ProtocolBeingModified: - Page 4. 6. ExperimentaDlesign:A. Animals
- (2)Strain/Source
Reason for Modification-:To correctlyidentifythe nomenclatureused for animals
----receivefdrom CharlesRiver Laboratories,Inc. Modification: Replacethis sectionwith the followingchange:
(2) Strain/Source Crl:CDO(SO)BR/CharleRsiver Laboratories,Inc. Hsd:Sj)ragueDawley SDO/Harlan Sprague Dawley, Inc.
(G21/01-07-91) Study DirectorApproval:
29