Document 936kvnG33pjkDkwN13R24O2Zq

AR226-3178 HaskaenlldLInabdoursatrtioarlyMfoerdTicoinxeicology June 11, 2001 TO: ilot Developmental Toxicity Study in Rats cc: L. A. Malley FROM: AJ.nCa.lyMtiacsallaCnkhaemist ANALYSIS FO DOSING SOLUTIONS MHAAannesaadklliyyecttlaiilclcSaaRllaemRRseeepfapleerorcrNehtnuNPcmeruo:bmjeebrc:etrN: umber: . NSeortveibcoeoCkoRdeef:erences: H24A7-628001-022 The,analytical report for the study identified above is a summary of the mixing and stability for th e jk flH IH H p m d will be used as reference for all studies under this MR. nfoe ' Pilot Developmental Toxicity Study in Rats ANALYSIS FO N DOSING SOLUTIONS Medical Research Project Number: HAnasaklyetlilcSalamReppleorNt uNmubmebr:er: SUMMARY HA-2001-022 pDroespianrgedsoalnudtiocnoslleact tceodnfcoernutrnaitfioornms iotyf/5co.0n,c1e0n.t0ra,4ti0o.n0 vaenrdifi7c0a.t0iomn ga/nmdLstoabfiilipty| ^an^a^lyHsisB(>Jnpoeurre "f Mf MrloiaoamrrmcchBhte71m,,m22p00e|r00Ba1ltuo^froocer!sruoiemnncgfFooelsrlboemrcluuitaettyirdoy/nco2son6na,thtc2ete0hn0feot1rsu.aartmtDihoeodnscavioynengnocfsfeionrceltaufrtart^iitogioneOnrsasnftriwMooenmarreatcnhphdere1asp0naa,mar2leye0dz0pea1rdne, pdfdaoocrrsoasitlntilaoegbcnitleoidtfyoonn rseoflruitgioernastifornomtotbhee asnamalyezperdepfoarrarteiofrnigeration stability/concentcraotliloenctaedloonng wthiethfo5u-rhtohudraryooomf temperature stability. In addition, 0 mg/mL (control) samples were submitted for analysis with the set of samples. used in this report refers to the active ingredient (a.i.) i Concentrations o^ chromatography [nseparate dosing solutions were measured by gas The solution vehicle for the study was de-ionized water. Runeisfuolrtms fitryo/mcoanncaelnytsriastioofnthveerdifoicsiantigonsoaluntdiosntasbpirlietpyaorendFaenbdrucaorlyle2c6t,ed20f0o1r, indicated that test substance was mixed properly, at the expected concentration ( 17%) in all levels and ptsshtoraaelbutpltateihroeeinndste-tahcsntoedlsvluecebcohstlieltcaedlnceotcenfedotwrho5aenshfMosotuuaarrrbtcshlheadtu7ar,nyod2ooe0mr0f1rthteefefmrosiregpueecrmroaantftiduooirrntemi.ofinrtRoysm./ecsoRutnhletcesseusfnlarttomsramfetoioparnnrtehavpleyeardsriaiofstisicooiannftgitoihsnneodldiucotasitoienndgs indicated that test substance was mixed properly and at the expected concentration p(foreup1rt1ahr%ad)taioaytnoaiflnlrdeleifcvraiegtleesd.ratRthioaentsutahltnesdtfetrhsotemnsuahbnesaltdalyn5scieshowoufartsshsaettadrboolsoeimnugntdesomelrupttehiroeasnteus rcceoonflrldeoicmtitoetdnhseo. nsatmhee as not detected in the 0 mg/mL (control) sample. 0 Compaq Safhfredi Boss not containTSQk CBf HA-2001-022; Page 2 SAMPLE SUBMITTAL All dosing solution samples were collected on the same day the solutions were prepared or as indicated by the stability analysis for the study. The solution vehicle was de-ionized water. METHODS 1. Analytical Methods a. Dosing Solution Treatment Samples containing^B lB B B W ftt the concentrations of 0, 5.0, 10.0, 40.0 and 70.0 mg/mL pwrosetrnaeferbrMpieigalairectarroycatl.ihtloeeSncd7at,we2smdt0eap0rbole1ienlsciFtfofyoelr/rlbocemruocunntaeitcfrhdeoyenor2mtsnr6aai,Mmttyi2oea/0cnrp0oc1rahnencpfa1eoal0nyrr,atsur2tianis0ot.i0ifno1oSnwramfvmoeeirrtpery4ilfc/e-cidoscoalaalnytetcicrotehetnnefe.dtrrisagSoatenamiromaMentpecvaldoerecsnrshictffaeri1bocn,iamtl2rtiiat0otyt0hin/o1ecnaosfnsnaodcwmre54eenr-htedroaacutyoirollnecatnedd 5 hour room temperature stability analysis. Eofacdhiludtoesdincgonstarmolptloe wanasexapneaclytezdedcoanftceerndtrialutitoionnowf 2it.h5m,4e.0th,aonro7l.a0nmd/go/rmaLn (eaq.ui.i)vparlieonrttaomount analysis. Samples submitted for analysis were analyzed the day the solutions were received or frozen until analyzed by the testing group. b. Chromatographic Conditions Instrument: Column: Injector: Detector: Carrier Gas: Split vent: Injection Volume: Oven Program: Initial Temperature: Initial Time Level 1 Rate: Level 1 Temperature: Run Time; Hewlett-Packard Model 6890 GC HP-1, 30 m x 0.32 mm ED, 0.25 /xm film thickness Split, 250C FID; 300C Helium (2.7 mL/min) 53.2 mL/min 1 microliter Gradient 50C 0.0 min. 40C/min. 300C 22.25 minutes HA-2001-022; Page 3 c. Calibration and Quantitation Aas satnoaclkytsiocalultrieofneroefntchee) twesatsmmaatdeeriainl (maestehpaanroalt.e BsaemfoprleeaonWalyBsis^oHfBthBe sHaBn i|eHs-c2o4l7le6c8teudsefodr February 26,2001 preparation, appropriate aliquots of the stock were diluted with the methanol to make calibration standards, which bracketed the target concentration of the diluted dosing samples. Before analysis of the remaining samples in the study, appropriate aliquots of the stock were diluted with the methanol after an equivalent amount of the diluted control to match the samples was added. This was used to make calibration standards, which bracketed the target concentration of the diluted dosing samples. Peak heights (4 retention tcsiqomuneacsre)enftrrreaogtmrioesnGssCiofonarn(daseloyessiFinsiggousfortelhueltsaioe-nssItadwnfedorarerdadserewteperrmreesienunestdeadtbivtyoeaccpoapnlliysbtirrnuagtcittohaneccpauelriavbkersah)te.ioigMnhctesuarsfvruoermebdy least rceopnlciceantteraitniojencstiaotnasllorfettheentsiaomn ptilmesetso(nea=ch4)caislibrerpatoirotnedcuarsvteh.eTcohnecmenetarnatrieosnuflot rfrtohme stahme ple. T(eCaec.sVht .sdu=obsssitntaagnndcleaevrudenl.difeAovrimactoiiteoyfnfi/inmciteehanent voxefh1vi0ac0rlie)aowtifoatnsheoevfmaleleusaasstutehrdeadbnyc1oc0na%lcceiunsltatrhtaietnigosnttashneidncaorddeufpcflriicictieaertnieotsnoafamvtpaHlreaiasstkifoeolnrl rLeasbuoltraotfotrhyefodrupaclicceaptetasbalme dpilsetsrifbourtieoanchodfothseintgesltevsuelbswtaanscuesethdrotougdheoteurtmthineestohleutcioonn.ceTnhtreamtioenanof the test substance for the respective dosing levels. aSstatbhielibtyaswelaisneevfaolrucaotemdpbayriunsginthgethcoermreesapnonodfinthgerdouopmlitceamtepsearamtuprleesanfodrrceofrnigceenrattreadtiorensuvletrsi.fication Company Does not contain TSCACB1 HA-2001-022; Page 4 ANALYTICAL RESULTS A. Chromatography ___ sluted from the GC column as resolved peaks over retention time range oi7n.f9ta,hp8ep.d2ro,oxs8ii.mn5g,aatmenladytr95i.x0t.omR2i0enpumrteeinsseuwnteteasrt.eivFdeeoGtretCrhmecihnpreuodrmptooastbeoegorrfaemqpurseasanerteinttasathtiioovnwe,nothifne Figures 2(a - c). Test substance was not detected in the 0 mg/mL control. B. Concentration Verification and Stability Samples Analytical results from dosing solutions prepared February 26, 2001, and March 7,2001 and analyzed for concentration verification and stability are shown in Table I and Summary Table 1. The following table summarizes the results for uniformity/concentration verification and stability analyses for the February 26, 2001 preparation. Preparation Date 26-Feb-01 Nominal mg/mL . 5.0 10.0 40.0 70.0 Measured3 mg/mL Average % Nominal CV % Stability6 % Nominal %StNaboimlitiyn6al 4.37,4.23 8.40, 8.39 34.6, 32.0 61.0,61.7 86.0 83.9 83.3 87.7 2 0.1 5 1 83.7 80.5 79.0 84.5 100.2 93.1 94.5 93.5 ba DStuapbliilcitaytesasmamplpelseshealndalfyozre5d.hours at room temperature, c Stability samples held for 4 days refrigerated. The results for samples prepared on February 26,2001, show that the test substance was astdaebqleuaitneltyhemviexheidcl(eCwVh'senlehsseltdha5nh1o0u)r,saattthroeotmargteemtepdelreavtuelrse(atanrdgfeotr=4+da1y7s%reoffringoemraitnedal.) Aanlld wrmsetaioptndhodirifntaieredddsv1tbao0le%uiinneocsglffupondrroeempthmaiernaseatedrli.sxianTmcmoepsrelrtteehsscuatanbiorosentlalwnfoocwirtehetwhroeutahts4at-nnhdoeaetyxmdpreaeettfcerritcixegtdeecdroaanritnrdeedtcwhtsieaoas0mnd.mpulTegehs/tmoeaLnmthdeseatahamllonpradelleysw.utialctsaslare Sw no* ah' esc HA-2001-022; Page 5 The following table summarizes the results for concentration verification and stability analyses for the March 7,2001 preparation. Preparation Nominal Date mg/mL Measured8 Average mg/mL % Nominal CV % Stability" % Nominal Stability0 % Nominal 7-Mar-01 5.0 10.0 40.0 70.0 4.72,4.54 9.36, 8.75 40.2, 38.9 63.2,61.7 92.6 90.5 98.9 89.1 3 5 2 2 101.2 107.4 97.0 97.2 105.2 103.3 120.3 99.6 bac DSSttuaapbbliiillciittayytessaasmmamppllpeelsseshheeallnddalffyoozrre44d,ddaayyss rreeffrriiggeerraatteedd., then 5 hours at room temperature. The results for samples prepared on March 7, 2001, show that the test substance was teasleetxdamvpebeqeplleucsetariieatnndetultt(yhhr1eiem2s. v0isTex%ahehmideocplfr(leeCinnswoVgumhl'wtseinfneloaerhresl)estahlbtdcehuca4t7en0pids.t1a0ad0ybum)sl,eergaetf/tomfotrrhsiLgateh,emter4aap-r4tdlge-ianeddytgaeayrdnaendrflerdeffivgoraeielngllrsoaealwrt(yaetteatdiedrcdgsabaelsymtvta=a5pbrlihielaoi1btiuys1ir.l%histiTygao.tehfsreAotnroslotlumhmobatisnhntaeanrl)ceand was not detected in the 0 mg/mL sample. C. Conclusion mReixsueldtsprforopmerltyh,eaatntahleystaisrgoeftethdelesvamelsplaensddsutraibnlgetuhnedsetrudthyeincodnicdaitteiotnhsatotfhtehetessttusduyb.stTaenscte was substance was not found in the 0 mg/mL samples. Cofl'erlVSsr*teed ses xsca>cS? I*' HA-2001-022; Page 6 ACKNOWLEGEMENTS Samples analysis by Sheila A. Riley, (Chemistry Associate) and Janet C. Maslanka (Chemist). SIGNATURES Report by: Date issued: * JaneCthCe.mMisatslanka H- AcO i Day/Mo/Yr *0* ion'nl * HA-2001-022; Page 7 Table I. Concentration Verification and Stability ol Preparation Date _________ m g / m Sample Type Nominal Concentr2a6ti-oFnebV-2e0r0if1ication Control - 0.0 5.0-1 5.0-2 Mean: 10.0-1 10.0M-2 e a n .- 40.0-1 40.0M-2 ean : Dosing Solutions l J ^ ________ Mealiured Percent Nominal ND 4.37 4.23 4.300.1 C.V.2%o 8.40 8.39 8.390.01 C.V.0.1% 34.6 32.0 33.31.8 C. V. 5% -- 87.4 84.6 (86.0) 84.0 83.9 (83.9) 86.5 80.0 (83.3) 70.0-1 70.0M-2ean : 61.0 61.7 61.40.5 C.V.l%o 87.1 88.1 (87.7) Stability 5 Hour Room Temperature 4-Day Refrigerated 55..00 4.18 5.01 83.7 100.2 5 H4-oDuaryRRooemfriTgeemrapteerdature 10.0 10.0 8.05 9.31 80.5 93.1 5 H4-oDuaryRRooemfriTgeemrapteerdature 40.0 40.0 31.6 37.8 79.0 94.5 5 Hour Room Temperature 4-Day Refrigerated 70.0 70.0 59.2 65.5 84.5 93.5 W Duplicate samples per level were analyzed. Mean, S.D. and C.V. calculated to verify uniformity of mixture. ((((ECDB)))) MMDMeeeenaaaonnnterrreeessssnuuuollltttt ooodffeftdttehhuceepteoalidlrcl.iagatinenaaslluybasenmsailftytoesrdisthsaaenmdduptplhelesicoraertpiegoisrnataemldp.slaems.ple reanalyzed. Company Sanitized. Does nnt contain TSCA CB1 HA-2001-022; Page 8 Table I. (continued) Concentration Verification and-Stability Preparation Date Sample Type Concentra7t-iMonarV-2e0r0if1ication^) Control Nominal "" 0.0 Measured n d (b ) 5.0-1 5.0-2 Mean: 4.72 4.54 4.630.1 C.V.3% 10.0-1 10.0-2 Mean: 9.36 8.75 9.050.4 C.V.5% 40.0-1 40.0-2 Mean: 40.2 38.9 39.60.9 C.V.2% Dosing Solutions Percent Nominal -- 94.4 90.8 (92.6) 93.6 87.5 (90.5) 100.5 97.3 (98.9) 70.0-1 70.0-2 Mean: 63.2 61.7 62.41.1 C.V.2% 90.2 88.1 (89.1) Stability 4-Day Refrigerated 5 Hour Room Temperature(^) 5.0 5.0 5.06 5.37 101.2 107.4 5 Hou4r-DRoaoymReTfreimgepraetreadture^) 1100..00 9.70 9.72 97.0 97.2 5 Hou4r-DRoaoymReTferimgepreatreadture^) 40.0 40.0 42.1 41.3 105.2 103.3 5 Hou4r-DRoayomReTferimgepreartaetdure(C) 70.0 70.0 84.2 69.7 120.3 99.6 1S5 Duplicate samples per level were analyzed. Mean, S.D. and C.V. calculated to verify uniformity of mixture. ((BC)) DSaemnoptleess nhoetlddefoterc4tedda.ys refrigerated, then 5 hours at room temperature. Compir sanKfced. Does not contain t$CA CB1 HA-2001-022; Page 9 Representative AnFaliygtuirceal1Calibration Curve Figure la:rseoCplualiltciiboartneastpiooqfnalkcfulhlreMvigehH(tRHm.TeM.a=sud7ri.e9lmumetenintdsuto(esvsqe)ursaahreocswo)nifncogernlctiarnaleitbaiorranftiirtoa(nnlignee)otfo Figure lb:tsooClruaetlpiioblinrcasattieon^caukrvheei(gRh.Tt .m=ea8s.u2^rimelumtienedunottevsse()rssqahucoaowrneiscn)egnfoltirrnacetiaaolrnibfirrtaan(tligioneneo)f 1.579 to 6.58 mg/mL. Compara? Sanitized.Dots noi eotrtarWCA OH HA-2001-022; Page 10 RepresentaFtiigvuerAe n1a(lcyotinctainl uCeadl)ibration Curve Figure lc: rseoCplauliltciiboartnaestipooenaM^cuhrevMigeh(Rt--m.Te.a=sAu8ri.e5lmumteeinndtusto(esvsq)eursaharoecswo)ninfcogernlcitanraleitabiorranftiitroa(nnlignee)otfo 1.579 to 6 .^ m g /m lT ^ ^ * Figure Id:tsooClruaetlpiioblinrcasattioeojfn^lgHcaukHrvheHei(gRBh.THt m.d=eba9si.lu0urmetmeindeuntotesvse()srsqahuocaworenisnc)egfnoltirrnacetaiaolrinbfirrtaa(tnliiognnee)of 1.579 to 6.5? mg/mL. Saprteed. 0*10 HA-2001-022; Page 11 Representative Gas ChrFoigmuarteo2graphy Chromatograms Figure 2a: Representative GC chromatogram of 0 mg/mL (control) sample. Retention times used foi re approximately 7.9, 8.2, 8.5, and 9.0 minutes. FIDI B, (010312JM008F0801.D) pA 20 n ilML I I ti I Figure 2b: Repre2s.5en. tative GC chromato7.5gram of nominal concentration of 2.5 mg/mL. 5T.0hme gm/iiienasuj rpedMconPceHntrMatiornloosfindgi1e7s.5oreluptrieosnendt_ia2lQuti_tveed stooluation is 5.36 mg/mL. FIDI B, (010312JM\005F0501.D) Wli l j l.liJi UP Figure 2c: Representative GCchromatogram of 3.29 mg/i solution. lyt1i7c.a5l reference Cotnpsrty San' s-tco,,*-fain TSeACBS HA-2001-022; Page 12 Table 1 OUimildlV Ui L/uam ti OUIUUVU ruiaxyjvu Sample Type 2/26/01 Concentration Verification Average Measured Conc.c Average Percent Nominal0 Standard Deviation0 Coefficient of Variation0 Stability Dosing Concentrations and Stability r ------- M ig /m L ) Nominal: 5.0 10.0 UTo 4.37 8.40b (87.4)a (84.0) 4.23b 8.39 (84.6) (83.9) 4.30 8.39 (86.0) (83.9) 0.1 0.01 2% 1% 34.6b (86.5) 32.0 (80.0) 33.3 (83.3) 1.8 5% 61.0 (87.1) 6l.7b (88.I) 61.4 (87.7) 0.5 1% 4.18 (83.7) 5.02d (100.4) 8.05 (80.5) 9.3 ld (93.1) 31.6 (79.0) 37.8d (94.5) 59.2 (84.5) 65.5d (93.5) 3/7/01 Concentration Verification Average Measured Cone.0 Average Percent Nominal0 Standard Deviation0 Coefficient of Variation0 4.72 9.36 (94.4)a . (93.6) 4.54 8.75 (90.8) (87.5) 4.63 9.05 (92.6) (90.5) 0.1 0.4 3% 5% 40.2 (100.5) 38.9 (97.3) 39.6 (98.9) 0.9 2% Stability 5.06e (101.2) 9.70e (97.0) 42.1e (105.2) 5.37f (107.4) 9.72f (97.2) 41.3f (103.3) a Numbers in parentheses are the respective percent ofnominal values. MMeeaann,rSes.Dul.taonfdaCll.aVn.aflyosredsu(pnl=ic2a)t.e samples calculated to verify uniformity of mixture. SSSaaammmpppllleeesss hhheeelllddd rraeetffrrroiiggoeemrraattteeemdd pffooerrra44tuddraaeyyfssofaron5ldlhosowaumerdsp.lbeyd.5 hours at room temperature. 63.2 (90.2) 61.7 (88.1) 62.4 (89.1) 1.1 2% 84.2e (120.3) 69.7f (99.6) qo8s t\ G1 $ti k*a*ed C6'