Document 0gJNDB51e8GrwMG1YEORxGzwx
oo
| AR126-0007 =
AbsorepvteironaofsiFngel-e252O-r4Macs D1onsreata
6
cotober 26, 1973
Conducted at: Durtig: Conducted bys
Report by:
Reviewed by:
RSSiuekbesripdauiLlaa,rbyoMraoitfnonreaisneost,a
Inc. 55101
une to uly, 1979
5. 3. Gibson and 3. D. Johnson
faanmeisorD. Wlnehmassaon,nt1&Pharmacy ologSz iastte
|
wh
ManaegerT, Dhruegr,haPtahbo:iisa Sata
000056
te Se
i
3.
Suemary
Asfotleurtiaonstionglgeroourpaslofdostehroefe mFaCl-e95r-a"t%sc, (amteanledaosste,95%4.o2fmat/hkeg)totianl cfaorrboenl-i1m4ina1tsiosnysotfemitcoatlallycaabrsboonr-b1e4d fatro2m4phloausrmsa.isTh7e.5hdaalyfs-.life
000057
Satzotuction
"
t
o,5fo-s0; xt
7055 *banotes Fosition of Camben-is
7SSrFh-ce5ac5rTpapltetaearsimontaFeh,neecplssoinhtmaaaisnscsaribteeuitemonniaopnCnlFdasnRsnOaoe-tfd9a.5vtpe5FerrremofnmimeuetscctoeeitipBfneoaatdneyey.dTtoossuPsiOfiLcon3ortSnmescssbpeaeeceorirndksp.seioofSnAa.LkrsecSerrHiieFneeosessiInagnieon
Ea(rnFeaChr-poEexlerSrsaEimnHaeeannhnteiemi13)neanThrrhieiiorccnihhoadvnveatatssaa.dvSaeosnssiiiomsenenesapnsseesdfpooovepperrnoovvHiiscdoEeeP,2doeavtfeeaoplomenecotahbSoeaErsesitosevftsosmxatnrosrpctrxiconnres
OCSFhoiomeSiconikanatalinotsnsa,mobanonnsa1h4es"xsepxorcveteteiitnoiinoenS:biitsatTwryialnslsafdoibrveimoasnte,ieodntthooe.fseScncFieEar-p)3acsetboesmooertptrchioninnss sDotneemns
ethos
hRSatedriucocaleraabbroeenl)-e1d4FsheLea-b9ep5le-c15ic5elaet tshcexisvoistyitoifonthaist.ototrheosfulFfeuirssaggale. (Ri(vSeere sShootvaeee STIriiLnanLvgaSeecAnibtoTonSr,ycsNCoruhpmembmaeiertcaanlb44oe2tc)ihbeernieescspt0air.s4ii5ts9;aitiuoCanis:/tmagta.inad oTseFhdeitohPoeCh-e9sm5pi-ei1ea9ttCeipweausstottfsiouuamnidtyetSoteebnern"e sina MSFaoelnteaauCdhraefrtrlaoebncso2lh4Rio3iovmieSorsrc?agSreC1os5D0,trFaonntras,aw4n em2hi0og5uh0trt,sowremheaekiesororletdca,ssdewweeoemrrnneaicAoilnlhdeioetnibootnosensdeSteoeraiennndeividual vostng Sach non-fasead cat use veishod ssmadiately before being siven a single oral dSS1ohi.soL2ea1ilveomrgfnetsFFuCCbw--ai99tt55i-h-o11n44aCc./4c2a0v.e0T.hcmeol.GhdloaassTesho.wevasiysanevise2nr.eas0nge,meilu(dtFooiefsovenioavnwsea0ess.)9p%4r.He2NipaomeCegrlm/eskdsgSo.ibltyuhtTaihboednsidecoroseneiteaw2inaonsssin2g0 seofesaanFctCn-fea9ornr5mi-cai1ao4nsrCeidntSgohbayk0Sao.rL9B,si%ttieNaocannkCd.lc,coonsTmtheoeaikniFcgoangrgib(nofsgneo:srLeAopnPpeceoornhdtasielunfnp.ehs1oomufartatnhaett smceooemdiernrgaatsesmosrvpoeteeidonsin
2 CPhuarrilneastaRnivCerhoB,reeRdaitsncgonLabPourraitnoariCeosm,panWyi,lmiSnv.gtoTno,utsM,assM.issouss.
Aloe Medical, Division of Brunswick, St. Louis 3, Missouri.
000058
5.
Sample Analysis for Carbon-14
Haormtosgenoifzwiantgeorf (fwe/wc)estoanodnetipsasrute owfasbdioolnoediicnaWlamraitneariballe.ndeTrhsebhvoamdodciennaatnesinweere
wteaircihnegdthientcoonceomabnudstaidodnincgone1s.?0aionf trthieplhiocmaotaeenoantea. toCoa-rleoadwiasnatbaakleanncteo
by mix
t(hferohmomtohceen2a4teandbet4w8eheonursapmoolsitnadso.se Uarrionuen,s) rweedrbeloaoldsocmeellass.uraenddipnltaosmcaomsbaumsotlieosn
cthoenes24bayndwei4a8hth.ourThaeroucoosncweanstrdaettieornmionfedcabrvbobno-t1h4 cinombbulsatsimoanfoarndtdhiere3ctratcsounitnina.
dPilraescmtalysabmyoletsran(sifnecrlruidnigna1.t0heml24oafnpdl4a8smhaouarndpo1s5tmldoosef Aorqouuaosso)lwerteo caouscnitnetdilla-
tpliaosnma,viaulrinaendancdournetdinbgloiondacellilqusiadmpslceisntiflrloamtitohne s24pecatnrdom4e8tehro.ur pHoosmtogednoastee,
gofrocuaprwbeonr-e14comfbroumstbeidolwoigtihcaalPascakmaprldesMowdaselde3t06erOmxiindeidzebry. coCmobmubsutsitngiobnlraenckovfeercyal
hdoomsoignegnastoelsutiaonnd
baltantkherbeedgibnlnoiondg,celmlisddlsep,ikeanddwietnhd
dilutions of FC-95-1c of the analysis of the
exper-
imental sample set (see Appendix 2).
LAlilquriaddiSocmienttriilclamteiaosnurSepmeecnttrosmewteerre. donFeorupsliansgmaa
Packard samples
Model 3385 Tri-Carb counted directly, the
esafmfpilceienacnyd froercouenatcihngs,ampcloerrweacstindgetfeorrmibnaecdkgbryouandddinfgrominatpeprrnoaplrisattaendbalradnktso,eaancdh
cfoormpbaarciknggrotuhnedrewsituhltsapptoroparisaetaelebdlasntkasndaanrdd ofofrkneofwfinciaecntciyv,ityt.he Acfaterrboc1ondrrecting
fcoornteenatchofsaemapclhe wsaasmpldeetwearsminceadlcublyatuesde.ofFotrheoAxiEdSiz(eAdutsoammaptliecs,ExtceorunnatlingSteafnfdiacridein-cy
wzaastioand)derdattioosmeeltehcotde.d sTaomplceaslibfrraotmetthehegerxotueprnoaflssatmapnldeasrd(,thirneetewrintahl lsotwanAdBaSrd
arasteiaosledandstatnhdraered.witDhathaigwherreaticoosl)leacntdedthonesepunscahmplteaspewearned rpercoocuenstseedd baylontghewith
CDOaCta17r0e0duCctoimopnutetro Sdypsmtweamsinaccthoempl3ishCeendtwriatlhRethseearBciholoDgaitcaalProAcuetsosmiantgicLEaxbtoerratnoarly.
Standardization computer program.
sample Collection
Groups 96, and
o1f44thhroeuersrpatosstwdeorsee.sacRraitfsicweedrebyaneexsstahnegtuiizneadtiwointhatdi1,eth2,yl6,eth1e2,r
a24n,d
48,
bfleorordedwatso darahwenparfirnoimzetdhetduebsec.endPilnagsmaaortwaasopfreepaacrhedraptroamnpdtliymmbeydiacteenltyrifturgaantsi-on.
dIingeasdtdiivteiontratcotppllaussmacaonndtenrtesd
blood cells, (esophagus,
sttoomtaaclh,urisnmea,lltoitntaelstfiencee,s,
spleen, large
tihnrteeestirnaet,s ainndtchoelo2n4),`aanndd48remhaoiurnsdeprosotf cdoasrecasgsrouwpesrefsoravecdarbforno-ml4eacahnaloyfsitsh.e
a Packard Instrument Company, Inc., 2200 Warrenville Rd., Downers Grove, Illinois. b New England Nuclear, Boston, Mass.
000059
:
6.
Results and Discussion
hanealyrseessultasreofshotwhne uirninTea,blefec1e.s,Thdeigedsitgievsetivteracttrapcltusandcocnotennttesn,tsancdonctaaricneadss odnosetheatav24erahgoeu,rs3a.n45d%3O.f24thaetdo4s8e.hourTsh.e beAatn24fahcoaulrse,xcrtehetimoenaniss1u.m55o%f otfottahle Scoamrebono-f14thiisn fSWecelsikealnyd dreipgreessteinvtestsryacsttepmliucsallcyonatbesnotrsbeisd c5a%robfon-tlh4e dporsees.ent eaitrheesrultinoftheexcdriegteisotni.ve Tthreacdtattiassfureosm otrhein48thheoudrigpeossttivdeosetrgarcotucponotfenrtastsasare cFoCn-s9i5s-t1e4nCtdwoisethwatsheab24sohrobuerd fporsotm dsoosleutdiaotna.afteTrhusa,dmiantislteraastti9o5n%toofnotnh-efasted rcaatrsc.ass.TheThmeajocarrpcoarstsiodantoafartehenortadaisoarcetliivaibtlyeraescotvheereodtvhaesr tfiosusnudeidnattahesince wlaasrgeQifvfoilcumuelthtoomoagsesunraet.esThewreereinsecsoemsesareyxcarentdihoonmoogfenteoittayl ocafrbsoanm-p1l4e ianliuqruoitnse o(f1-t2o3t/aalayc;)arbtohnis-14iaafitnerconantraisvtdotsoethoef mFiCc-h807s-l1gyCer(r0a.t0e67o8/fdauyr)ina(r1y).exTchreetion scpalrebeonns14frocnonttehnet,24anhodurtheanpder4c8ehnotuorfpotshte ddoossee rinatsthweerwheolaenaloyrzgeadn wfaosr *tot0a.l2%. pTohsits iisv dmousceh lfoorwerFC-t8ha0n7-1t4hce 6(%1).of the dose in spleen chserved at 124 days pTaheredcon(cTeanbtlreati2o)n.s Tohfetmoetaanl rcaatriboon-o1f4reidn brleodobdlocoedllcetlolsplaasnmdaplcaosnmcaenwterraetiocnome aatfte2r4 aandsin48glheouorrsalidsos0e.25ofaFnCd-905.-3194,c,restpheecrteivieslyn.o seThluesc,tivaet r24etaenntdio4n8 hoofurs carbon-14 in red blood cells.
T1h,e2,res6,ult12s, fr24o,m a48n,aly96s,isaondf p1l4a4smhoaurssamaplfetserfarosminggrloeupsoraolf dtohsreeeofratFsO-a9t5-Lc adraetashisownplointtTeadblien F3.igurTehe1.logThoef mleeaanstcosqnucaernetsratliinoen vtehrrsouusshtitmhee ifnodritvhiedsueal Points from 24 to 14d hours for these data fits the equation:
Cp = 15.657:%9%7%_ is plasma concentration. The hale-life of elimination carrobnonp-l1a4smaaftiesra179shionugrlse o(r7a.l5 ddaoysse).of FTChu-s9,5-1e4lCimiisnatsiloown. from plasma of total
References 1. JoofhnTsootna,l JCDa:rbonE-x1t4entinanRadtsRouatfeteorfaExSicnrgelteionIVanDdoseTisosfueFC-D8i0s7t-r1i4bCution
(Report), September 29, 1979. 2. JDoohsneso(nR,epJoDr:t), AJbusloyrpt10i,on1o97f9.FC-807-14C in Rats after a Single Oral
000060
:
7.
List of Tables and Figures
Table 1: P#eCr-c9e5n-t14RCectooveRraytsof(MTeoatnalDosCea,rbo4n.-214mg/akfgt)eraat S2i4nagnlde O48ralHouDrossePoofst NDBose5.2584 p. 7.
Table 2:
PCloamspmaarisaofnteorfaToStianlglCearObraoln-1D4oseConotfeFnCt-9o5f-1R4eCd
Blood Cells to to Rats (Mean Dose,
4.2 mg/kg) at 24 NB 52584 p. 7.
and
48
Hours
Post
Dose.
Table 3:
CDoonsceenotfrFaCt-i9o5n-1o4fCCa(rMbeoann-1D4osei,n
Rat 4.2
Plasma mg/kg)
after at 1,
a Single Oral 2, 6, 12, 24,
48, 96, and 144 Hours NB 52584 p. 10.
Post
Dose.
Figure
1:
Mean Oral
CDoosneceontfraFCt-i9o5n-1o4fcCa(rebaonn-1D4osei,n
Rat 4.2
Plasma after mg/kg.)
a
Single
NB 52584 p. 10.
TApapbelnedi1x:, NDBete5r1m31i2napt.ion35.of Carbon-14 Content of Dosing Solution.
ATpapbelnedi2x:, RSepciokveedrywitohf TECo-t9a5l-lC4acx,bon-14 from Blank Fecal Homogenate Samples NB 52584 p. 12.
ATpapbelnedi2x8,; RReedcoBvleoroydoCfeTloltSaalmpClaersbonS-p1i4kedfrwoimthBlaFnC-k95F-e1c4aCl, Homogenate and NB 52584 p. 8.
Appendix, Table 3:
Comparison of Oxidation with from Rats at 24 and 48 Hours
Direct Counting of Plasma Samples after a Single Oral Dose of
Fc-95-ldc.
NB 52584 p. 11.
000061
.
8.
Table 1
Percent Recovery of Total Carbon-14 after a single oral Dose of PC-95-14C to Rats (Mean Dose, 4.2 mg/kg)
at 24 and 48 Hours Post Dose
-_--
Fercent of Dose
Rat
PosTti.nDeose TcaTrcass hTiragcetstpilvues Feces Urine
Number'
(ours)
Contents
-_------
1
2
77.342
3.27
1.68
1.67
2
20
77.06
3.45
1.72
1.04
3
2
82.62
4.02
1.2 1.09
Mean
79.01
3.58
1.55 1.57
a
a8
103.61
3.55
3.25
1.01
5
a8
86.21
ERY]
3.10 2.78
6
a8
92.63
3.24 3.37 2.88
Mean
04.15
3.32
3.2
2.52
_--
2 pata were adjusted for combustion recovery (see Appendix 2).
000062
-
i
5.
Table 2
CFeodmpBalriosoodnCoefllsTotaanld CPalrabsomna-1a4ftCeorntaensitngolfe oral Dose ofatFC-2945a-1n4dC48toHoRuartss P(oMsetanDDoossee, 4.2 mg/kg)
N-- umRbaetr -- PoreTimHeoe-- s R-- adSBhleood -- Plama-- RaMtaitoio-- nreedd Blood
terre em----reeste----
1
2
3.6302 16.342
0.22
2
2
as
14.62
0.28
3
2
3.18 13.40
0.24
Hoan
0.25
4
ae
2.80
10.27
0.20
s
a
7.85
10.25
0.55
s
a
4.94
1.7
0.42
Hoan
0.3
_--_--
2 Data are expressed as ug equivalents of FC-95-M4c/g. b The data were obtained by conbustion and were adjusted
for recovery (see Appendix 2).
000063
;
;
"0.
nb 3 Concentration of Caxbon-14 in tat Plasma BE after a AST Single Oral Dose AA of FC-95-1 A4C i i (Mean Dose, re 4.2 mg/kg)
N Tooorusre x a5t F 00 zSTveworre
3 :
na se
aw 1am
rez en
oox2.n20 1x33
. uw
na wa
ous es
12.60 sna
w11.a98 s2 0n.55
2 w
eas we
ies ase
13.02 11.06
160 12.89
2.60 2 1.60
5 10a
11.08 on
ser sss
1100 000
2s0.57m20o.78
2
Three rats were dosed
ata were obtained by
dainrdecstacrcioufnitciendg:fodrateaacahretiemxeprientsesrevdala.s
ug equivalents of FC-95-14C/ml of plasma.
000064
, 3
x :
:;
3
6
3
E
:i
4)
2
3
f
.
on Conomnsation ot casheneia 9 sat phases After a Single Oral Dose of FC-95-ldc (Mean Dose, 4.2 mg/kg) "ee nssquares we ty 9 rs 5 a 3
ours 203 sone
000065
Co
12.
Appendix 1
Determination of Carbon-14 Content of Dosing Solution
Just prior to dosing of rats, the FC-95-2C dosing solution was sampled
with calibrated micropipettors? directly into counting vials; six 10 ul
and six 50 U1 aliquots were pipetted. One ml of water and 15 ml of Aquasol were added?, corrections were made for background and counting efficiency and, using the specific activity of C-95-1%c already deternineaS, the uCi/2.0 ml was calculated. The data are shown in Appendix Table 1. The dose administered each rat was 2.0 ml of solution containing 0.553 UCL of PC-95-24C which 1s 1.21 mg of rc-9sc-. This is a mean dose of 4.2 mg/kg.
22/1 wicropipettor, Lab Industries, Berkeley, California. Badquuriansgolspewcasififcounadctitovibtey daetseurimtaibnlaetiosonlv(etnot-bescirnetpiolrlteendtsefpoarraptce-lsys)-.ic sbpeecriefpiocrtaecdtisevpiatryat=el0y.)4.59 uCi/ng. (Specific activity determination to
000066
yo
1.
Appendix
Table1 Caxbon-14 Content of Dosing Solution
yCTiofL2M.0img1 r
.
0.5602
0.5554
0.5614
0.5628
0.5612
0.5554
5y0Ci%/2.A0limg1wt 0.5626 0.5532 0.5290 0.5390 0.5302 0.553
Overall average = 0.5526 0S.55e3 ci/mg = 121 HCL FC-95-1Y/2.0 m1
000067
re .
Appendix 2
He
BlRoetcoosviecraylofSamTpolteasl CSapribkoend-1w4ithfrFoOm-BB5l-a1zikC
Tshaempgloeosdharsecboeveenryreopforttoetdal (c2)a.rbonS-i1n4ceftrhoem ccaormbbounstleadbeFlC-i8s0a7-tCo tihnebsiuollfougrical aetxopemctintbhoatthgoFo-d95re-c1oCverayndofthetotcaolmpcoanrebnotns-1o4f Ffcro-m8077C--19c5,-14ictainsarberasointasble to metabolites in biological samples can be attained. Por each set of samples
combusted, four replicates of 10 ul, 50 ul, and 100 ul of diluted re-95-Yc
dosing solution vere aliquoted with calibrated micropipetiors directly inte
scintillation vials. At the same time using the same solution and pipettes,
either three or four replicates of 10 kl, 50 ul or 100 ul were aliquoted directly into combustion cones which already contained blank biological material (fecal. homogenates, plasma, or red blood cells). The combustion cones were dried and then pelistized vith 5 on ashless filter paper. Blank filter paper
pellets were combusted and the solvents collected in the vials to which the FC-95-1%
had been added directly. One each of the 10 ul, 50 ul, and 100 ulre-95-Mc
spiked pellets were routinely combusted at the beginning, middle, and end of each set. of biological swmples. After correction for background and counting efficiency, percent recovery was calculated by comparing mean results from direct addition and combustion. The mean recovery data for three sets of
fecal samples that were analyzed on different days for total carbon-14 (pc-95-14c)
for Fo-Bxperinents Xto be reported later) are shown in Appendix Table 2A. The mean recovery 1s 98.0 + 1.5K. Since each entry in the table is the mean of four determinations, this mean recovery 1s based on 3 determinations. Thus, good
recovery of total carbon-14 fromre-95-Yc spiked biological material is shown.
The Bean recovery for the set of sales in the report (FC-Experiment 4) which
is based on triplicate 10 ML 50 ul, and 100 ul FC-95-Y9c spiked blank fecal
homaenates and red blood cells vas 93.6%. (See Appendix Table 28). This slichtly lower recovery vas due to a minor malfunction of the Packard Oxidizer and as fairly uniform throwhout the set of samples. Thus, the total cazbon-ld
000068
Appendix 2 (Cont'd)
I
1s.
data obtained from combustion in this report were adjusted for recovery.
There was no apparent differences in recovery between spiked blank red blood cells and spiked fecal homogenates.
000069
wo
:
16.
Appendix Table 28
FecalReHcoomvoegreynoatfeToStaamlplCeasrbSopni-k1e4d wfirtohm BFlCa-n95k-|,4c
_-- --1_0 md o7.e 100.6 97.9 98.6
50 u1
l100m u1
99.5
97.7
97.0
98.1
95.4
28.1
97.3
98.0
_-- Overall average = 98.0 + 1.5%
2a amount. of Fc-95-11c reference soluti. on added.
b Data are expresssreadtiaase \ recovvereyry 3GEPRmIIfOrRocmomcuosmtbiuonstion
100)
Each entry is the mean of 4 combustion recovery determinations.
000070
-
7.
Appendix
Table 28
Fecal HRoemcoogveenrayteofaTnodtaRledCaBrlbooond-C1e4llfrSoammples Spiked with FC-95-19c
--_-- DIRECT ADDITION
10m
HomFoegceanlates 661193 677
X66
1002 756%. 756 727 123 741 + 18 Red BClelolosd 6741, En 691
501 36388688
3719 367 3737 + 90
SPIKED AND OXIDIZED
501
Fecal
Red Blood
Homogenates cells
33578751 35358833
3484 3347
3613 3504
100 1 7403 --740g4 7394 7400 + 6
100 1
Fecal
Red Blood
Homogenates Cells
77140088
7093 7098
7105 7081
7106 7091
S6734a61% x 100 = 85.3% 8$9315 x 100 = 93.33
3$651535 % 100 = 96.78 335034 x 100= 03.80
T71o0e6 x 200 = 56.08 T70a01o5X 100 = 95.8%
_--
Overall average = 93.6%
2 amount. of c-95-19 spiking solution added.
2 bata are expressed as dpm.
sample was not used; cracked vial. 2 Spiking error; sample was not used.
000071
"i 18.
Sppentiz3
ofCoPmlpaasrmiasoSnamofpleOsxidfartomionRawtisthatD2i4reacntdCo48unHtoiunrgs after a single Oral Dose of rc-95-1%c
Concentration of Carbon-14 in plasma for the three rats in both the 24 and 48 hour post dose groupwas determined by both oxidation and direct counting. Samples were counted directly by transferring 1.0 ml of plasma and 15 ml of Aquasol to scintillation vials and counting in the liquid scintillation counter. Corrections for efficiency and background vere made for each sample. For determination by oxidation, plasma samples were measured by weight into combustion cones and carbon-14 was determined by combusting and counting. The data from the plasma oxidation - direct counting comparison are shown in Appendix Table 3. The mean ratio of oxidized to direct counting is 0.97+0.02. Thus, the plasma level data obtained via either of these two methods will be essentially the same. The dizect counting method, being much more convenient, was chosen to analyze the plasma, and the data quoted for the plasma concentration versus time (Figure 1) is from direct counting. Though a direct measurement of absolute recovery from plasma was not done, these results indicate that recovery of total carbon-14 from plasma spiked with PC-95-19C would be similar to that found for recovery from fecal homogenates (98%, Appendix 2).
000072
yo
:
19.
Appendix
Tabte 3 Of PCloamnpaaraSi'asSmoipnnlgeolsfeOfOxrrioadmlaRtaDitoossnewaitotfh3F4eD-ai9nr5de-cl4t48cCHoouunrtsinagfter
RatTTT CoEunteing
Oxia dation oimaatmion,
21 ((2244 hnoouurrs)) (u1g61/.m8134)53
a/9e
f1r5i.e30s
Direct Counting
0o.ls97
13. ((4284 hhoouurrss)) 11336042 2 (48 hours) 1358
1311235 1333
o0sss ols
3 (48 hours) frees
10
Too
s ess r ms--X-- +8800..9977-- $400..0022
2: ata axe expressed as ug equivalents of Fc-95-2ic.
.
000073