Document q3YEzJEz5Vk90DQZZy4dqyooj
AR2RG - 3268
450
AR226 - 3268
4
HaskealnldLIanbdoursattioarlyMfeodriTcoixniecology
March 13,2003
To: C Study in Rats.
RepeatOrealTdox-icDityoGasvaege Rane Finder
ce:
FROM:
G. A. Ladics
J`A.nCa.lyMtiacsallaCnhkeamist
ALYSIS FOR H-24691 IN DOSING
IONS
~
MISXITNGAANBD ILISTTUYDY
REVISIONNo.1
Medical Research Project Number:
Haskell Sample Number: AAnnaallyyttiiccaall RReefpeorretnNceu:mber:
Service Code:
`NSottuedyboNoukmbReefre:rences:
L2I46L91Y
H24A691 2001-037
tached is the analyticaluetpmoartytboesuastiesdfyfoprroottohceorlpruerpqousierse.ments for toxicology studies with
Company Sanitized, Dass mat contain TSCACE! ~
RHAE-V2I0S0I1.O0N57N;o.P1age 2 --~
SAMPLESUBMITTAL
`Samples containing H-24691 at the concentrationsof 0, 1.0, 5.0, and, 25.0 mg/mL were collected `VoenriTfeisctatDiaonya4n. dT5h-ersoeomsatmepmlpeesrawteurreeastnaabliylizteyd. tSoadmeptleersmifnreohmomtohegesnaemiteyp/rceopnacreanttiroantwieorne collected ohnoTmeogsetnDeaiyty11(1a.f0tmerg7/dmaLysloevferlefornliyg)e,ractoinocne.ntTrhaetsioensavemrpilfiecsawtieorneaanndarleyfzreidgteroadteedtesrtambiinliety along with S-hour room temperature stability after refrigeration.
Aorllatdotshienegstsaubslpiesnhseidosntasbailmiptlyetsimweerfeorcoalnalleycstiesd.oTnhtehye swaemree adnaayltyhzeedsuwshpeennsrieocnesivweedr.e prepared
`The suspension vehicle was 0.5% methylcellulose.
METHODS
1. Analytical Methods + Dosing Suspension'
HEa-c2h46d9o1siinngtshaemspulsepe(n3smioLn). wTahseddiolsuintgetdsoa1mp0l0mesLwweirethanmaeltyhzaendowliatnhdomutixfuerdttheor ddiilsustoilovneotrhe
~
`owferaepfpurrotxhiemradtielluyte0d.0wi3tmhgt/hmeL0(maig)/pmriLosrtamopalnealGyisniist.alBdefiorelalutl oitnaaniledxoipleuntcitoe)nds,ctohneceinnttreamatlion
sstaamnpdlaerdtdoigliuvteedasofilnuatlicoonnc(ernetfrerattoioCnaolifbarpaptrioonxaimnadtQeulaynt0i.t0a4tmigon/SmeLc.tiEoanc)hwsaasampdldee(dftionaelachtest
dilution) had an equivalent amount matrix (methylcellulose in methanol) when analyzed.
`Samples submitted for analysis were analyzed the day the suspensions were received by the testing group.
b. ChromatographicConditions
CoInlsturmum:ent: DIentjeeccttoorr:: CSpairtevreaGta:s: IOnvjeenctPiornogVroalmu:me:
IInnittila TTeimmpee:rature: LLeevveell 1| RTaetmep:erature: LToetvaelru1nTtiimmee::
H3e4l&We,nDPB-a1cM7okd0ela16083,r9203mGd0mCImD,,1umfilm thickpess. SFpIlDi:t235500CC H2e3lmiuamin(2.1 min) 2Grmaiicrcoaltiter 110000mCin. 220060ClC in. 0900mmiinn.
--
REVISIONNo.1 --_ Be AZULIBTIUED esti
c. CalibanrdQauatntiitaotinon
A separate sample of H-24691 was obtained to use as the analytical reference standard. A stocksolutionwaspreparedinmethanol. Thisstocksolutionwassonicatedtoassurethatall `materialwas in solution. Beforeanalysis,appropriatealiquotsofthestockweredilutedwiththe. 0mg/mLsample(initial dilution)tomakecalibrationstandards,whichbracketedthetarget `concofethendiltuterddaositngisamoplens. Astocksolutionofintemalstandard (1H, 1H,2H, 2H.perfluoro-99-8% mpuree)wtashpryepalreddienmecthaanonla-nd1ad-dedotolea,ch calibrationstandardandtestsampletogive afinalconcentrationof0.04mg/mL.Theratioofthe internalstandard and H-24691peakheightsfromreplicateGCanalysisofthesesolutionswere sedtoconstructacalibrationcurveby leastsquares regression (seeFigure 1forarepresentative: curve). Measuredconcentrationsforthedosingsamplesweredeterminedbyapplyingthepeak heightratiosfromreplicateinjectionsofeachsampletothecalibrationcurve.
~~
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CanpanySanizD0e08dT.tontain TSCACB!
REVISION No. 1
-
ANALYTICALRESULTS
A. Chromatography
Hap-p2r4o6x9i1maetleultyed3.f5rmoimntuhteesG.CTchoeliunmtnemaasrlesstoalnvdeadrpdeealuktewditfhraoemtethnetGioCnctoilmuemonfas a rcesohlverd peoakmwairtahesahtroewtenonitingoFntigirumreeoasf2(maap-psr)ox.iTmeastetlsy 3.u9 mibnuwtsaess.ntoRetpdareetseencntteadctiinvetehGeC: 0mg/mLcontrol.
B. Mixing and Stability Study
Aannaallyytziecdalforreshuolmtsogfernoemitdyoasnidng/osrucsopnecnesinotnrsatcioolnlevcetreidfiocnatTieosntaDndayst4abialnidtyTaersetsDhaoywn11inand Table and Summary Table 1.
The following table summarizes the results for all homogeneity and/or concentration
verification and stability analyses.
STaemsptleDayTyp Noomeml _ Mewuneg d TMB' Mea%Nno(miTnMaBl) CV9.) %SNaobmiilntayl
--_
"Test Day 4 Homogeneity
o
ND
--
~-
5100 054a4d,0n84843,40m83t6" 980328
64 s95s0a
TestDay 11 3 marae 920
5 %0
7DayResfuibgielryied! 0
Not
-
--
5100 0900p,08r64y,0859 5074
30 0read
2 2,239
964
1 pr
&b FSiaemapnlreeshseulsdfSorhohuersaatlyosoimsotfeimperoaptu(re).,middle(V 3d oto (B) amples.
@ DMeenaontersesoulnteodfaeltlcatneadl.yses reported.
Meanresultofallanalysesreportedexcep forone duplicatereatsamplewhichwaslow dus10alquot
MDiucpalnicraetseulctoonfcdeunptlriactaitoen rvee.rsiafimcpaltiinognsfarmopmltehseaonrailgyizneadldeixlcuetpetdsfaomrphlee.1.0Omrgig/imnlal.aneavleylsi(shwomaogselroncwietsy)t.han expectedduetoaliquot errorandnotreported.
gy i. 0455 1otcontain TSCA
~~
=
RHAE2V0I0S1I.O0N37N;o.P1age --~
``ThhoemdoagteanfeoorussalmypmliexsecdolilnetchteedvoehniTcieestatDaalyl l4evienldsi(caCt.eVs.t"hsa=tt6h,e4taesntdsu5b,srteaspneccetiwvaelsy). The testsubstance wasatthetargetedconcentration inthesamples ( 11.2%of nominaaln)d was stable in the vehicle when held 5 hours at room temperature. `tThheetdeasttasfuobrsstaanmcpelewsacsohlloemcotgeednoenoTueslsytDrae-ysu1s1paefntdeerd7idnatyhseovfehricelefart ailgleievnerdlisac(aCtt.eVis.t'ohsant= o3,f0naonmidna1l,)reasnpdecsttiavbelley)i.nTthheetveehsitcsluebswthaenncehewladsaretftrhigeetraartgedetfeodrc7ondcaeynstrfaotliloonwe(d+b1y2S.6% nhoomuirnsaalt)raopopmeatermepdetroatbuerlee.sss"TthaebSl-ehtohuarnseaxmppelcetfedobruthteth2i5smcagn/bmeLalttervieblu(t8ed4.to8%soamfpling and analytical variability and not stability of the test material in the vehicle. Testsubstancewasnotfoundinthe 0mg/mL samples. C. Conclusions c`Doautladfbreommitxheedanhaolmysoigseonfetohueslsya,mpwlaessadtutrhiengtatrhgeetmeidxlienvgelsstaundydisntdaibclaetuentdheart tthhee cteosntdsiutbisotnasnce tnheececsosnacreyntfroartmiionxsintgesttehde.doTseisntgssuubsspteannscieownasstwnioctefwoeuenkdliynftohrea0nymgst/umdLy wsiatmhpilnest.he range of
ei
--~
raevro e Pp
ACKNOWLEGEMENTS
`Sample analysis by Sheila A. Riley, (Chemistry Associate).
SIGNATURES
Repotty:
Date issued:
(oCWc alent ta) Chenin Janet C.Maslanka
13 Ho = 2003
15-Ham2005.
Day/Mo/Yr
ps
tain TSCACB!
~~
`iCompany'Sanitized. Do8S'notcot
HAR-E20V0I1S.0I5O7;NPaNgo.1 e 7
--~
"Table I. Homogeoeiy snd SaboiflH-2t46y91 inDosingSuspensions
Swe
mgmiHael ___
Pawm
Type
M ommlewwd Nomi!
"teow
`CHoomnogoerne.ity
000
NDA)
-
MoTwore Boru
1100
ooss s
98444
110010
00880690
08690
102 mean(): 008843602003
@38..0 6%
Mean): 05c8v8.:e00%5
(8.8%)
MoTooLr z
5500
aas3
4586
Borrow
550010
a41m6
984342
502
521
1054
mean(): 4.7240.56
(94.4%)
~~
Mean(): 4c5v1.2019
90.2%)
MTooor s
22500
2227
89083
Borrou
20
2
95
Mean): 2c3.v04s10
(92.0%)
`stabilief)
100
osis
ss
1100214
00888533
88833
1928
0522
822
50
mean(): 0.48155410.04
(895.540%)
20
20
9%
B(bc) TD oocaivem eeranalgoefomfvtheieaoaionsf eaprl.iiquleote0d,)amocddbooppoelrnncce(nctroelfeiveogmuntin,aralo=(mi)tihpeexooerrpgitennfaoltr,i2t5emeadngsva.dmepdlveeveri1so,i2gon)o,.1e0d=D).
8)) MSeilmtysamtplresihgeld Ch0o)rs oao.moeemi er lossddp ly ofbesnd
Eres
~~
company Sanitze Do8 101
RHAE2V0I0S1I.O0N37N;o.P|age8
--~
`Table I Homogeneity and StabilityofH:24691 inDosingSuspensions(omtinved)
TT T ed wgmLB PewI
`SampleType
Nominal Measured Nominal
Tos Day 11 "
ConioL.
0
NDA)
---
MpTooirz. Bottom Vera
" " "n
1100
0056040
89604
10 Mean):008857942002 (8875.49%)
cv.3%
50
446
22
50 Meanp):4n452001 (898..08%)
cv.o%
2200
2239
9958.6
Mean): 24c.v1.0.12%1
(96.4%)
Si
7-DAYREFRIGERATEDYS HOURRT 10
~
10
009s55
895552
Mean): 0c.v90.4a20%.07 90.4%)
7-DAYREFRIGERATED! HOURRT 5500
442118
88466
Mean():42c1v:.01.%06 (842%)
7-DAYREFRUGERATED/SHOUR RT 250
205
820
20 Mean():212.280.92 (s50782%)
cv.a%
(8) TWheoavmereagemessurdconcentron,averagepercent ofnominal(inparentheses)standard deviation,ancoeficient
() Tohfevaavteiroagneomfeopa,mdidceon,cmeidatbiootni,oamv.eragepecentofnominal inparthses),standarddeviton, andcoeficent
(D) OToFhfevvaaivaerrtaogneoomsffeddauusppulltiricceaadticeeoossnaacmenmpiceasi..oMne,aavnereagselpeorfcdeunptloicfatemri-naanlaliynspsaorfenthheesoersig)i,naalnsadmaprled.evOiragtiinoan,l0sdrcolefaniacleynstis
was owe hanexpectedpeoprted.
~~
RHEAV2I00S1I.O0N3N7o;.P|age --~
Figure 1 `RepreAnaslyteicanlCatliabrattioinCvureve
12 Equation for the fitted line:
109 Y=0012550 + 23.0624 * X
08 R-Squared = 0.9968
2 06
5
= 0s
4
& 02
~
00
000 001 002 008 004 005 Concentration, mg/mL
Figure 1:C(aslqiubarraeti)foonrccuarlviebsrhaotiwoinnsgolluitneiaornsfoift l2i4ne6)9r1epdliifcuaetdepoveearkchoenigchetnrtartaitoimoenasruarnegmeeonfts 0.t000.01468m5g/m6L.The0mg/mL. controldilutedsamplewasuseda th ro.
orcontatnTEER
Company Senfized Hoss
~
RHEAV-I2S0I0O1NoN.P.0a|g3e71;0
~
Figur2e
{I Figure2a: RIeFprLeIsFepLn2tea.tri2v5feGuCochrrooma.tmogeratmohfy0mlg1d-a0eac(dcdaoenndr.l)RsaempltewiteitmhenfottrenF-ai2s4to6a9n1dniarsd Spproxim3.a5mtineulsy.Reteton timsforintemalstandardis approximately 39minutes. -
--~
=
=
2
Figure2b:RceproesnentcateiveonGf0rC.0ca3hmrtogmo/atLno.grTahmeomfe2a5smurpedmconHc-en2t4r6a9ti1odnoosfinthgesroelputrieosnentaitliuveedso0lutinoonmisn2a4l.2mg/mL.
[IEE Figure2c: RepresentativeGCchromofa0.0t312omg/gmLr.1a246m9)analyticalreferencesoon.
--~
CompanySanitized. Doss ot contain TECACBI
REHVAI-S2I0ON0N1o-.P0a1g3e71;1
--~
TABL1E
SUMMARY OF DOSING MIXING AND STABILITY ANALYSES
DosingConcenoftH-r24a69t1(mig/omLn)
`HomogNeonemlitnyaSla:mples
10
50
250
TestToDpay 4
osu
443
22
044"
(#86)
9
Midd
oss
a3
27
@84)
4)
08)
Boom
0836"
an
22
6
44)
68)
AverageMeasuredCone." oss
ast
20
`Av`eSrtaagnedPaerrdDceevnitaNtoimoinnal
20808
20021)9
2+100
CoefficlentofVariation"
%
a
5%
5HSotuarbiRloiotmyTSeammppelreasture
oss4*
475
20
=
50)
60)
TestDay11 7-DayRefrigerated) Top
0500
446"
22
00
2)
68)
~
Middle
0864 (864)
a9s
5269)
Botom
0859
---
--
(159)
`AAvveerraaggeeMPeearcseunrteNdoCmoinnea.l"
o@s7.40
ass. 0
21 64)
Coe`fSftiacnideanrtdDoefvViaartiiaotinon
2002 3%
001 03%
soz 1%
5HourRoomTemperature
0504"
an
22"
04)
2)
8
+MNoebneresstpofathreoersgaaeaGliequrostpsedcidvueplpiecracteenatloifqupoosfraolmvhles.giadiedsample.
4SMtoaatnorebsasteodogahelaavieroagee,smoenaesduurpeldiccaotnecaelpiqtuiootnd(prm)eofshseoofpt,hiedselceooadddubpliicoateoaflicqauocth.dosing evel
+DMueplniceatlecoof dnuplcicaete narmaltoof nrvireraploirstecmd eoax.noionrmstytofplinsg.
--_
os
la
RHEAV2I0SLIO0N1N:o.F1uge 12
`Reason For Revision No. 1 `Thereportwasrevisedtoreflect achange inFigure 2 (page 1o0ft) he report. The changewasto
c "Gao s Cthr hroehmr eaatdoe egrraac nphdyatlClhrroemafteogrrtaemons"cHoriegGshC"PerfLo iqur idCmhra oman togc rape hyorHPLCt"o
SIGNATURES
Reportty:
CoCPo clant ker _ 5Mor-soos
Chemist Janet C. Maslanka
Day/MolYr
~
_
Company Sanitized. Does ot contain TS!TSCACS!