829 



1«6 



8YH0 ET AL. 



giucutonidt and '"•f'lc^aminopbcllyl jlucuronislc uiine Iherrao- 

 tpmy l-C/MS oocrai«l under (lie conditioiu descnbed in itus 

 jcudy. Howcvee, ihe ebility of {hcrmotfiny LC/MS to d«t«ci 

 glucuronidi: cuaju(pU3 Is vjrtiblc (UiO very Jependeni on ihc 

 druciure of the islycfinr ( I S) Diract determi nation of 'Jic eJu- 

 curonidCT may have to woii until a yynthctic Qtaadard ia evaiUbie 

 for ociimitation of ihc ihermospniy system. Glucuronidcs of 

 Icrt'ory Kro""''": MTiinc« und »lipK3ti; tertiary aminui have been 

 detected in urine By LC/MS. but only if^cr itic avtILibitity of a 

 synthetic siatidard (19. 20). Wc are currently working on the 



rjolndon «nd cHaractcnmtion ofthase 6luciimnid««. 



KyeremJien « al. (9) reponed the eicretion af glucurontOe 

 oonjugitac of nicotine in tmokefs fnllowine inlfivenous ddtnin- 

 iitraiiun of "C-lst>=led ratxmic oicoline. The jlucuronidc ob- 

 scrwxl in that study was wtrfomioinlly (/aw-.^'-hydro^ycoiininc 

 glucuroiiide, u jurtj-id by «ppc«r»n« of fro« '"C-labeUd irant. 

 3'-hydroxycotinine (bllowlo| i[KUt)atlon wtUi d-glucaronid«3e 

 of Ihe polar HPLC fraction ceotaining the putJtive tlucuroaidc 

 peak, llic finding that /moj-B'-hTdnwyoofanine was coojogflletl 

 10 the greatest extent is ouzdins. considering our findiflgs «nc) 

 Curvoll'e fndinjs. Howev«r, i( mun be rerocanbcred ibil K.ycr- 

 onaten adtniDistercd a very low dose of ratJiobbcled nicotine to 

 abttaining cmoker^ It a poulblf that, uoder coaditioos where 

 Uje circulntlBg levels ofcotininc hire been reduced by »b«enllon 

 frora Mnokittt the creater Droootlion of « iow intrivenous dose 

 of nicotitje ij ooovcrtcd to (njfff-J'-hydro^ycotinint jiucuronide 

 rather than cotinine glaciLfoalde. Altenutivdy, because :0% of 

 (be Qiootiae admioiuerod "m the K.ycremittn ihjdy wms JH+) 

 nicotine, the tK-itiWlity custs that R(f-) niwtioe ia metabolized 

 to the gluojronides ai s dtflisect rate thin the 5f-) tuootlne. 



Without c»ceT>lion, unooajUBai«d t«iiu-3'-hydro!tyootiDin« i» 

 •he tna|or urinary metabolite Ibr all anoken In this sttKly. 

 constituting in average of 35S of til metiholiiei meawred 

 (lnclt)dlr.g the conjugated fbnns). Althotigh Gr« reported many 

 yean a£o as a nicotine metabolite <R maniinala (21X lrJ/t^3'- 

 hydroxycotiiune proved diflkult to quantify in urioe beeauae it 

 was Qot readily vKUtaai baa-dse of its hydrophilidty. The 

 tbsrmotpray LC/MS method tited in this ttndy d<iei not involve 

 cxtreolon and I: wefi-«uited for (he dltta detfirminatidc trf 

 urinary minj-l'-h>«droiyMtinine. Pbannacolonetic studies of 

 rranj-3'-hydfoxyootifunc have shown that it is foraMd from 

 cotinine and fiirther metaboliim Is limited (22). Thus, it Is no? 

 jurpriiiiij for it tn be ooo«i«t«ntly found 3 th« major urinary 

 meaboltte. 



[ndividutl respoQsss in nicotine metabolic profiles rrilect io- 

 icrestiiTG di lTei r .iiu a. Smoker 2, fot exatnplc. Is unujual ccFm- 



poKd wjh the oihcn. in that mucli taore free frow-J'-hydrox- 



ycottnine, cotinine, and nicotine «ur« found ia hit urine aod much 

 less of the cotinine glucuronide. With the exapuon of Smolca 

 2. tht .-di'.ive p.-oportior. of nicotine estccetad ss each nxsabotite 

 5ho^vJ variioom b/ factors of 2 to 7 o«rr tlic iO reoiaiaias 

 irroirefs. -Smolcef 9 has no deiectaWe amoant of the nicotine 

 glucuronide, birt rr.or^ than twica th« amount of co<inirj<-AJ- 



o;(ide than any other sraoter. Thete oftservailons sufflest that 

 certain metabolic path* ays may be favored ovcrcihcrs in some 

 suiolccts. 



The ciata -.n tabic 2 rcaardint smoldnfi behavior cf the indiwd- 

 uiUs in tlii.» study an: limited in their ability to d«»crib« the actual 

 amount of moolinc aotortied try each smoker. The FTC Itself 

 hw rfoop"i'«d that FTC ra(«d "milliftrara nicotine per cigarette" 

 ^.ilues do '-01 necissanly represent the actual njcatinc vidded 

 iliirinj imoldiij b\ .iiiy pnrticular individual (23). Thus, the 



agmber of cigarette* end (heir tot«l PTC nicotine yield provide, 



at best, only a general sense of nlattinc absorption for these 

 Rmolccrt in a cteady-ttau- jyrtcm. However, a major objactive of 

 this study vvu to quantify the tvtat nicotinc.<tcrivcd materiel 

 eicreied in urine over t 24-hr period for smoken dunng ihdr 

 tcjulMT cmokj<»g routine. Thut, the data in table 2 serve to 

 cuiifli'iK ttut Uv: subJocQ arc regular smokers wtio consume 

 between one-hall and four o«oks of cifireltcs per day. 



Although (he nicotine oul,'in nxka in table 2 vary approxi- 

 mately 3-fold (0.5? to '. -M) fur these smosars, the dau Indicate 

 that, on average, 9654 of the FTC nicotine yield can he recovered 

 In urine as nicotine equivalents, based on the anil^-tca determined 

 in this jtud>-. We have not used the tho-moapray LC/MS method 



to quantify tite amoum of nicotioe equtvatenu in humaat ad- 

 minlstcrw! a tnown amount of aicoUae Imaveoously, tut wc 

 do know iSat gl% of ("CJoicotine adr.iinittered intravenously 

 cao be rcco>^rcd in the unnc aa mctftt^^licca that cou dc detected 

 by thermowray LC/MS (9). 



Summar]- 



Thermnrpray LCAIS has beso sucoestfiiJIy aDolied to survey 

 urj nary nicctine Tnctaboliic distributioa ia 1 1 tmolceia. For thee 

 smokers, the variability in ibsotute amount among the nine 



Boalytca ranged from }5 iu 70%. Thia mathod ooniiima the 



prtsetia Of guctjmnide conjugates of nicotine, cotinine, aod 



Iraiu-J'-hydroxycodnirvc, conditute an avera^ of 79% of the 

 urinary metabolites. trans-J'-Kydroxycotininc in tb: fnx fijrm 

 cQiutitutes the largest single metabotite in tmolcers' arioe. with 

 aa avcraj^e of 35% of the latal The sum of nicctin* mataboUtec 

 detenninod here is very close to lis STC yields furthe dgaraijea 

 smoked by thcuc xnbjeul&. 



{tefdcaccs 



1. G: A. Kytnntun, L. H. Taylar. I O ilHVfhir/, aed E. S. Veadl: 

 RadiooKinc-bi^ pofunnascc liquid dsraraatosrapby uay ^ 

 nicotine and twayr of lU cietiCOhtet J. Ctiromaiogr. Btamd. 

 .■(ppi41f. 191-205(1987). 



1 J. D. delUlliizy, M. R Bai«miing, Q. i. Doolitlla, J. R Robnteoa, 

 K. T. MCManus, C A. FUlia- R. A. Davis, O. T. Duiser, J. R. 

 Hiye>. J. H. Reynoldi, ijid A. W. Haycr Chetnical lod biolopsai 

 tuuit«i of a ci^fxue that beats nUicr tbaa bom tobaoco. / Clin. 

 rhtuTnxai 30, 75J-'63(i??Oi. 



3. J. E. Heanioffidd, C. COtien, aQ« O. A. caovjncc Out laxtx 

 constittmoa affect ^ obieeiive' diatnoeit of isjootine depend- 

 ence? ,</n. / Fvblk fJfoUk Id, lOiO-tOai 0900). 



< OB. Nctitalh aad F. O. Pric; <hu ehrOTn«<o«™pt»c defcrmiaanon 

 of rAiut;-3'-hydtoaycotininc, s tntjor tnetaboiite of oieotine Ir. 

 luiaten. / Oirumatojr. Olameii. Awl. <1S. 4(XM06 (1987). 



5. a B. Ncunik, M Dua^CT. D. Otth, ead F. G. Pein; /<i»i.»-.T- 

 H'jtlnrrycotlnlnc as i main metabolite In urtac of soiolren. Arch. 

 Ocav. t/jTirwi Utaihif. 199-20! (1981). 



<( M. r-jcall, E. K. Velt, C. Enjiuad, «no C. R. Eozdf. Unniry 

 cxcictiOD of oicotinv and it* mijor nirtaNstitca. ■l3rd TotM«o 

 Chemists" Re««rch Contkreoc!, Rjduooad. VA. October 3, i9S7. 



7. N. J. Haley. Y. Loitrinar. C M. Axchid. aod J. P. PJctiie, Jr.: 



Co"lribuliivi of »n*M-J'-hy<iroiy<Miinin« eod iluouraoiije oodja- 

 ^to of nicmiue aieialxitiiea to UK mMsijrett;cni of axjatr.e Itj 

 RJ A. Imematioail Symposiura «o Nicotine, Hanbuflg, Ot-inaay. 

 /uneiS-JO. I9«0. 



8. O t3. Oyrd, i'.--M- Ci«at, 1. M. Greece. B<vt I. D. deeethiiy; 



Detcraimatjon of nicotine and its mcubolltes In urtac try tnrr- 

 ntosnrsy I.C/MS. "If Tohacio Qierciits' Rraeirch Coaiwence, 



Wi„«,„-,<U«ni. .SC. S«pc«mbM- J0-O«nl-r J. 1940. 



9. O. A. Kjerrmiitn. M. L. M>)rfai.0.a-.«uci,T»(l/»r. / O. dsPed-'ty, 



anrl E S. VctKll: Darosiiiii.-i 3f r.iootmc »n<l elan meatxiUics In 



