rl .Rcpdrt Nr, 101 f L~rfRA~VRE ST~DI ~.Nilrosa~in~r ii ~obac[o Prodo:tr 'MAYIURQ O '3 U~P IM~IIOKLUIO ~1 'r e CI IbPD F3~p nf,,tl~:a~i~-e CON~fDENTfAL , ,,,,,,, ,,~; ~g iE.Report Nr, 101 E LITERATURE STUDY I·litrorlninrl in Totl~~o (todu~tl Project No,: 06,03 Author: p F1IStCr lulllrhed by: E. Klurch Septtmler 1986 Dlstrlbuti ~, , Hetrn Herter Herrn Rftterrh~u, Photocopy No,: 1 Mr, E,E, tdhnhar~t No,: Z Mr, A,L, Heard Or, R, B~nni No,: q.t Or, P,J, Door ~L~-_ )10~: 8 Or, S,R, Elorrey I No,: g nt, p,6. Nfchollr No,: 10 nr, I,n, oenton Dr, e,J,P, de Slpueirl " lol: 12 O I No,: 1J ru Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 TOIIErl O, Suml~ry 11 introduction 2, N·~Lrol~~ncl in Tnllecn P~duct~. I A tabutir [ompflitlan 2,1 Tol~co·S~cllle H~itroi~nlner lTSNAi B 2,2 Non·Vol~tile N·Nitroriiincr (NYII) 1a 2~3 Yolltilo N.Nitroiiiioci (YMli II 3, Summiry 3,1 Pra~ence of k.Nftrollmlner in Tob~cro Pro~uctr is 3~t In~lyiii Of I·Nltr3ll~inC~ in Toliecp Prolucli 20 3,3 Formation of H·Hltroramlncs In Tolaceo Produ~ti 22 3,3,1 Culttvatfon 3,3,2 Curing 23 3,3,3 Ferrcntition 3,3,4 Tolacco Conititucnt Subrlinrcr 2( 3,3.5 Iglng 26 3,j,6 5torige 26 3,3,t Production 3,3,8 ~ddftiver 3,3,9 Pactinp 2 I, Protpccts I, IftC'~ture 6, Annex EOP6EZLOP ClibPDF - v~~fastio.soni 1. O, Summary A literature survey piver a tabular rnPpilation of N·ljtlPldmi6F level~ in tobacco products since their d~icovery in these products in ]g14, The analytical data are arranged by typer :i N·nitroraminer and typer of tobacco, For a '(lnprehenrive ~iserrnent and cprnapricpn of the data, the tables 1;1! both the countries in which the analyser were cF Il~ctp~ as well as the analytical urhodr, rurlhermore, the ci~cilation also specifies the joutnalr in wh~C~ Ihr results were published. The data arp additionally iaentificj :o indicate whether they were origindll~ furnished by Hoffmann and eo·woileri (dHF, USA), or internally from the tilA~T Research LabPtatoriei tabOCIBATUKE RbO Southano:an (England) or R~~ BAfCT nalnburg (termdny), A survey lists all hitherto known interdependencies regarding incidence, farnalion and analysis 1 N·nitroraniner in tobacco produrtr, Finally, research progrdnmer Ire presented to gain further knowledge concerning a distin:: reduction of N.nitraramin~i in tobaccos, Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 11 rftroduCtign Tne first indications of the carcinogenic effect of I. nitror0·fompovndl (nitroran~iner, nitroia~ider) were erta~lirhea in animal eiperimenti some 30 years ago, Since then numerous scientists have dealt with this substance class, Iven teddy 3 to a publications on these substances are being published per day, Nitrosamines an their own are not carcinogenic sulJtanc:s, Oary in the mammal organism doer en2ymatit activation decompose them in:o aliylating netaboliter which react with biopolymerr in a similar manner as ONS, thereby ini~taiting the tunoropenic effect, Only ~aproiim~tely 10 X fo the more than jOO different I·eittoro·E0spounds did not have a carcinogenic effect on experimental animals, These included H·nitroloprPline (HPRO) which is contained in tobacco, A highly specific and ertr~,nely sensitive detector for N.nitrolo·el~poundl (ilR I Thermal Energy Analyrrr) developed in the USA some 10 years ago made it polsible to detect numerous exposition sources, In this context it is nPcas,ary to mate a distinction between in·vivo formation from nitroranine and njtrosamjde precursors (endogenic) and environmental pollution by these substance classes (eiogenic), Some of tnese results (rtudier onducted an ham, beer, medicines and roon air) have been presented by the mass media with the result that nltroiamines have become the focus of public attention, Press reports on tobacco products are also devoting increasing attention to N.nitrosanineS, Oruclrrey and Preussmdnn shiwed 1; early as 3gb2 that the formation of oitroramines in tobacco sno~e was possible in principle, But it was only in 1911 that Hoffmann and his co·workets were ~4le to identify an atkalaid nitrosamine, O namely I·nitrcr,nornicotine (NUNi, in tobacco, hi ClilrPB~~f;l~s~tl~c~:;i?~lil ·3· With the GC.IEA analysis system introduced for tobacco analysis in 1982 a total of 10 r.nitrosamines have since been discovered in tobacco products, the most recent one (kBAL) war discovered only a few months ago, the R·nittosamines identified in tobacco and smoke can be classified into three groups · volatile (I), non.volatile (2) and tobacco·sgecitic (5) k.nitroramines, ~ne latter are formed from the tobacco alialaidi and they account proporti· onately for the highest Concentration in both tobacco products and in mainstrem smoke, Whereas until now no N·nifr0samjnes have been found in green tobacco, there compounds can be present in tobacco products at concentration levels of up to 100 mg per kg tobacco (, ppm)'l toncentration levers ate particularly nigh In Chewing tobaccos and snuff, N.nf',rosamine formation is attributed to chemical, eniymatic and bacterial influencer during tobacco processing, particularly during curing, fermentation and aging, The following substances containeo in tobacco are primarily involved in l(.nitrosarnino formation: altaloids, proteins, nitrates, nitrites and pesticides, Approximately 11 X of the I.nitrosonarnicotinc (NHK) and approximately t :of the apen·ehain Nitroramine ketone (NNK) can transfer in a chemically unchanged state from the tobacco tu the mainstream smoke where they form 35 to 50 X of the total H~R and CNK content, the remaining nitrosamine types are pyrosynthetically formed during smoking, Since the nitrate level in ttbucco is gdTtiCU~1Tly important for nitrolamine formation in cigiclLte smoke, a significant reduction of this Compound class :I the mainstream smoke can be achieved by low·nitiate leaf and stem blends, a C: I i I; P B F-r:!!:!!:!:f3 Stl C:i: O'lil -- .4. The nitrosamine content of mainstream smoke can be reduced by as muCh IS 80 X by cellulose acetate fi)tCrs, and it can be reduced still further by filter ventilation, Volatile H.nitro. samines reduction is well above proportional, while for the to~aeeo·~pecific N·nitrosanin~r reduction is approximately proportiona/ to smoke dilution, the volatile ~·nittasamines contained in cigarette smolre were soon analyzed with the cC·~EA System installed in FE·ATD in 1979, Since 1982 additional tobacco products and mainstream smoke are being checked for tobdcca·rpeeific N.nitroraminer as a matter of routine, Furthermore, influencing variables are being studied wn(ch permit nitrosamine reduction (filters, additives, biotechnotolical processes), GR b DC has Seen operating a CC·TEA system to monitor this substance class since 19831 this literature survey covers the results on the distribution, formation and analysis of H.nitrosamines in tobacco products published in journals as veil as in GR b Dr Reports and FE Reports, the purpose of this assessment 15 to help plan and conduct further studies and measures to reduce N·nitrosamines in tobacco products Clit; PD F~ililil:f;l~t I'CTi'O'lil ~sr N-NI~ROSAMINES 1N TOBACCO PRODUCTS t TOBAt[O-SPECIFIC N-NITROSAMINES ('ISNA) NNN : II·HITROSOHORHICO1111E NNK : 4·(H·yIROSOH~,THWIHO)'1'(J'WRIWl)'l'sllIUIOIIE ~ NHAI: II·(N·H~tROSOMETHYUnIN0).~.(3'1YR IDYI)·I'WTAWO1 NAT r I·NI~ROSOANATABINE NAB t n·HI'TROSOAHABISfHE 1 2, NOI·VOLATrlE N·NI~ROSAMINES (NVNA) ~O E~A : H.N 1 fROS001 Ef D~O~AHINE ~PRO :H·N1TROSOrROIINE 1 YOLATlil R-NIIROSAMINLS (YNAI NDMA r N.N~TROSODtnl~THVUM~HE NUEA= N-NfTRDSODtE~TH~P~AHINE NDPA t N·NltROSOD1 PROPV1Anl~E ) NDa~; ntl(TROSODIBU~W~nlnE NPYA : N-Nt'lRDSOPYRRO1IDIHE NPIP ~N·nlTROSOP IPERIDINE NMOR : N.N I tRO SOMORPHOLI iiE O 4 C I i I; P·B F-r:!r:!r:!·fil s i 1 ) I . N-NITROSAMINES 1N TOBACCO PRODUCTS I~ IOBAC[O-SPECIFIC N-NITROSAPI~S~ 3 "~ HNN NNK WNI1 B I a n ii. N)B NONIVOLAI1E N·NltROSAtEIINES (NYNA) HO·CHp·CHOI H·nO 6 HO·CH~·CH1' NDEIA NPRO VO1AiLE NITRosAMINEs ~NA) f eHj~R'Na CtH6,H·Wo bnr, elHo, NNO N110 CHI e,n,' 5n; ecn,' HDMA NDtk #DPI HDBI (I Q O ,a ,a a IPIR I1I1 NL(OR j Si Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 j .I· 2, I.kitrord~incl It ~ob~cto Pioductt · I Idbuler taaptl~tfon j Thl follo~ing t~bul~r tomDiliti0n lilts the nitrosepline contents Of IOblCCO IrlduefS specified In publlcItl0nr end in ca r SC end lI Reports, fne vilucs hire been elAsilficd j primdri]y by the three Lypi~ of nitroIllnlneI (ChlDtlrr 2,1, 2,2, 211): i n Tob~~co·iperlfle I.nitroii~lnir (~SO~) 5 ' Zon.volatil~ I.nltro~~~loir (IYhl) r ' Yol~tlle h.nltroiinln~i (YCI) I Yithin there groups the v~luer ere ~rr~ng~d by tobacco type Or mdnicr Of ~onrunptlon: I(rudl Lobltro i~ir·, Iluc·, lunru,tl, lurlO, Yirginl~, Orient) B Cig~r~ g I pipe tobeeCo i ( Chering to5J:co I Snuff (snurf toblcro,'snuff dipping') ~urtnl~gning rlalllficlllOn r(thln thor lub.proup~ II pr 1 the cnployid ~n~lytlcit a~thod: b ,Ot (thin.t~ysr Chromltogr~phy) ,GC (psr chron~~ogr~pny) I HPIC thigh Fci(Orainte lipuld ehran~~togrclhr] I HPIC·~EA (kPLt therRleI cnetgY ~nblyirr) 1 ( tC·;EA (~t thCID~I rnergr In~lyi~r) An iGen'.ifier is plIccd in front of the Iltcr~turr sources ,, ;,,,,, ,,,,.,,,., ill .1. I·( ma III(U" and r~.~,r~erl IHol, frgn GILOC Soulh~ipLan tRD), 6 iron (C1 gur Ola In·h0ule Illni~ltidnl (/E)~ !iir c;alrification principle n14ei it porii5le to puIetly B difleren: tobacco pr0dretr, and to uo8ate tniir LIbICI bI nei results, R Jumplry O( Lhrle Inily~cl data II glvln In the :n~p~er~ J~1 and 3,2 of thir r1001t. CI h K q i~t .a. 9 1,1 IOBnE~O·S)IEIFIE N·HIIRD3UIIHIS (TSNn)l) gtohlrro In dry n~ltrr (pp) L1 (lit(tltpJI .:*L: NNN I NNK NIT I Nbb IfrSHI Ieet j Pi BHly, dr~c~Psdl US~ I,Z 1,1 oe Ho 10/19tl H1C 3 1,9 1,9 #~ I~Plcy,lrcen tab,l USI M I · I II WLC no Jpon " ,half-ury I " Jrr~4 ,OIlEd ,5.1,1 la,s~tlt I B~tlrl~ait·ewd ) US~ 1,7 1_1_1 (1,1 ~u~ullnm a ~ugrd ,~cd 5,1 I I I ( 5,1 Bplly IM 1,6 0,5 1,6 CC~U I Is 6911982 j Qly US~ ( 2,11 0,5 3,1 i · ( 6,( ) no 1/1983 IB~rly,gnen tob, ~'n· IM nn nn cr~e~i INo tlil#3 house) n ,f!uc~cur~d ,3·0,111,1-0,2 1 0,2-0,31 · 1D,FO,II " ' IflUC·CU~dIIfeno f 0,05 I~o,ol 0,06 I it 012 jt~i 'Is i~·~ll 0~1 o,~p,zl lo,s~o,( lir~wd ,e( 0,2 4,]1 0,2·0,4 led~ kly, namal '1,1 0,1 5,9 1·1 13 ~u Upo~ tar tlta: O,t 0,1 0,2 1~0 loid level h(g~ nr· 1 113 0,3 3~1 1 I Ib n(co:ine level an I not datrltable I) lie Ilgni~lon\ cnalytical valurr Irr ail~ble on the ~ so: e~s~ined fifth ;*SatcP·IDrciri~ I.nilMlmiLI dill (C6) II r 6?1:: aleinal(on 2) pr ' parts prr niltign: vs pCr p I ng pm lg toLwra p tl i ~E elJ·ination Ho t Holh~nn ct 11. 3) HLt I ho~ogrnilcd tuf·arilg ON *33~1- I .9. 2,1 fDBAttO·SPEtlFlt ~-~trROS~IIEI (TSIA) i Product ~ountryl NNN ~I Ilt I1I IL '10111 Illothod I Illt y9ll toblcrn In Iry nnttcr Ippn) lit,t~t~~ numuad 1# 11 I · I·I · I 21 02 11113 nlcotl blp~ 21 i~ II ~ I ~eoti low 90 I · I - I · I 01 no~icotl high I r 1 48 1 · I-I ·)I ~rtlco~ 1fumvsl,9rccni ~ Il · I·I · I r ,C2,DI I d ~uKurd,Lott~ P,Zt,~ · I I · utm 1~ Icll po~l I , tPP Icl,Y ·I · I · plCt~ d lnl ~~lt n~eurd I,O 1,0 1,3 3,3 B~(YI)O a Flumond le~ot~ 0,9 114 ) ·1],310~1EI~I 1/1JI1 ,~ lu91t O,I 0,9 2,9 ) " i IU91rl i I a,~ · I ~toutd h VLginil, jtCla 1~1. r I al I old. 1(E·ltl ID 11113 ,14,q 0,1~ 14,2 a ,ai·curedl · a I trrllr I 0,2 I< O,f 011 1 I IE 01110 1 O CllbPD~- v~~~asrlo.som - c ~ ' ~C~· 3 ·10. B a 3,1 fOBA6CO·SPEClfre N·HITROSAnlHES (TSHI~) '1 g Product Countr~ ~NN tobaco ln dry clttcrINI( Hlf WIII IftSHI, ILlhod I UOuettor ,Iluc~ o,( I·I · !· I oc a tlrm III cured Alrt I·I·I ',1 toblceo ,B1Lld 0,9 0,5 tl e~owtta I · I . 1154,al P Ilb nn I/i I Up~nrtsl OS~ ,O rPI Jt,M,OI a Jno 1W1914 1~ PIIM 0,3 re O,] ~ I*"" usn m 1,3 1Cltlll II ·II IClpv~t~' llSI ),O 9,0 tJ,ln s C~pmttd 1,0 m 3,2 Ila 10 1411~9 1: 0,2 0,( 0,4 rm 1,0 L (gtOndcl I,( 0,1 1,5 m 3,1 j; O,L d,l 0,1 m I I~ tutky Qgantn ,aland bennlny P,~t,l m CT~U lIE 05/1002 black 3,3 0,6 1,1 m 5,3 a eigu~ttn USI aSZn JHo 4111# O,f 1,1 Prince (O,b~ 1,0·2, Cipll~td, P41. 0,1 0,5 '0,5 ( · 1 1,3 1 oean IRoll~B~ I~ fluwsurrd US~ W,II1,CI,II ],w,O " ~Jll 0,5 0,2 0,7 1,1 6 Fnnce 5,2 0,1 1,1 7,3 Swftret 211 0,5 0,9 1,1 Cigucttb USI IE4U lbi(119111 Pql, 0,3 0,1 0,2 0,02 frlne~ ,C12 1 0,5·1,11 1,0-2,0 ctguetter uu ~,w~,l~,2·1,II1 2,14,31 · C,Z.i2,t 6·lu I a 1(11911( Clganttn ,81enll tcnany IS I OIM~ I q,)l ,O la 11,0·2 Ge-'IU I~E13/1#15 1,3 0,8 I 1 1,2 la ,aislt O,] I (D,1 Ytrglnl~ OrlCnt g 1 :~~==~=~~ 31 P~a I pi'ti Per nitlion I r919 'n9ltp tob~cto ClibPDF - v~~fastio.soni .19· F · tnd products contain differing I,nltrosanlnes Conecn· nine groups: tratloni, this opolles 1DDrOi(wtill to ail j nftrosa, I snuff ) Chewing tobacco ) Cigars ) Cigarettes tob~~Eo·iocclfic I.nftror~nfne~ arise in varying puantltles within the different tobacco products: o Cigarette : bt~ft, Burley ) blend ) flu.rurcd , Orient · Luin~ )) stens e grade aged > grade unaged n Lower leaf position ,upper leaf position ,ehewlnp tobacco : ffne,cut ) scrap leaf ,plug · Snuff I flnc.cut ) DPuCntl ) 1POIe leaf i : nolst ) bags ) loose loaf Note: Since the concentration levels of the individual N,nitrosanines In the tobacco products are not the sole conrideratlon, a classification of the W.nltrosao(nes by their biological activity (7) (56) is also given here, the grading, however, was derived froo animal (xDlria(nt~ and can only be 1 very rough guide because the results depend upon 1 nabtr of influencing variables (anlnal species, aanncr of application, dosls, period, organ specificity, etc,)l IDnA · very strong blologital actirftr kNl ;CMOR · strong carcinogenic hCI, NrrR, N311A · carcinogenic of wdluR activity Ilf, NAB , rc~~ carcinogenic NPRO · neither carcinogenic nor autagenic \D Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 1 6~11 b ·20. b 3,2 In~y! it of ll.l(ltrnrlninnr In 1Pl~rro I~olutti · The flrlt ~lil)Old nltroleleloe . I·nltro~0narnlC0tlnr (000) .wll Identified In tob~ctJ eI tite er 1974 (10), · nl~h·lcrlorm~nre If pull rhrPn~tolr~phy (Hlle) Ir cnnlorll to ~cp~r~tl E. end 2·l~ontr~ Of III end to d~tlrrln~ the Converrfon rite (Ch~wlng tobereo: II I E· end 2~ x t·nnr) (3), · Iron 1JIZ anrodl the eCIT[1 ~n~lylfr Lrllel Ill~rtl IIII~1 rgrld·ull~ fbr ~lllur,n(ntl, It het renIIne1 to till eer uorlv~li~d for It~ Ipcclllclty, Icnlltlvltl, riptoducl. blllfy ~nl !Oald, fhe rE Irlt~ll~tlon h~l teen wor~lnl k eery relllbly Ilnee 19)9, The CC·T[I1) ~r~~urlop nllhad Ir rupctlor to the nllt.TEAl) method for dctcrnlnlnp tob~c~o.rDcrlfle Ilnltro~~iln,i (TSMA): II II norr rrono~lc~l, fetter, ~o~e tenrltlve, end It II the onll Iithod thet r(ll trpirete Ilnllroro· In~blrlnt (11I) IrPn n.nttrollla~tibln~ (WIT) enl B Pit~mlne It 142, 56, bl), A fonperlIon 01 rilult~ rlv~~tl good eonlotnlty 121), a · A eompertlon betreen the ~r~~urfng Icthudl CC·tCA end GC.MPD Inltrogen pholohorul dbtlCtOr) rlVI11t thlt con~ldrribly h(ghcr k~nltroi~~lne vituet ore ab~ilned rith the triter brc~r~r It det~~ti further nttrogrnou~ iob~~to iubitencer In eddltlon to the I.nltro~~nlni~ (51), thC reErntly prerented EID(lllrY CC·~E~ Inllrllf trtten will IIPlt~tl end Idlntlly the volettle end tobiceo. Ipiiflle k.nltroi~~lnei with the tone eolunn, ~rl ~n~lytiC~1 pr0rcdu~( wel L(nptifiod end the dtt~ctlon th~erhold rll reduCed 153), B .lodcl rC1CtiDnl IndiCltC thlt I;tthCr P.nltr0l~nlntl Irl prrrent irl tob~ceo pr0duEtl, in eddltlun to the II Itr(edy (I!lb~ilhCe (1~), 911 lhrOn1109rlDh or h10h·D(t10tR1REI Ilouid rnra~~tsl'iPh rougled rlth the nltr0ll~lnl~ IDCtlfl( EltlCto' 14~~~11 lner91 Inl]rtlr (ftlJ, ClibPDF - v~~fastio.soni ; ~5~R 1211 .An ~i~o~bie aclalbuffer solution II nctirr~r~ for tobacco extraction to prevenl artefact formation (If, 1~, 55), for conDlttr extraction of tobacco produCts e lh~~lng period of et least id hours et room temperlture appearr to be In~rlt~bte ~19, 20110), · FE hei observed Cnnlbnlnation 01 the Column ~~t~rlil with COIZOZ·b~~rlng substinces when Extrelut colunns rrrl used for s/IP]e P~CPlrlllOll TLlt Is Yht F~ does not use ~ny extraction rolpPn~ ( opposed to Hoffnann end Co.r0rkcri and ' · Cdlclum hes been su99erled as reference substance to en~~ine the influence of tob~~:o Irritaent on the I.nitt0iialne level in tobacco oroducts, the pu~ntlt~tlr~ shire of these elements, for Inrt~nce durlnp eurlnp, remains unchanged (1), . the publlshet results were ptl~irltl furnished bl the wort tit notf~~nnllrunnm~on (UM). · The fl.nitrosemIne levels In clp~rittr end chewfng tobaccos, end In Crude end ~ni~f tobaccos, estibltshed by Ft for the berRin laerllet (C9·75) rent wftbIn the rctld·rldr Rleesured results, I~trhing Inunl) O1 tOblelP·IPC(iflC W.nl~rolrlnel In mainstream sAoLeI On th~ other bend the TSU vilues measured in tob~Eeo products bJ 6RIDC were In some ay FI. (ttJ, inn r~iion for Ihll II prg~~bly dllfcring instances substantially hljh~r then those esteblIshed sample p~eDSrltiOn (ba5OC uses SOirltt extrlCtlonJ (JO), O IVW Clil;PBF -!::!!::!!::!.f3 stlc.;: nill r5 i .22. If ' .1. jij ~otnltiDn of I.lltrol1nCI In 1Ob~ECO ~toductr 313,1 tultlvitl~~ · I·nltrolJmlncl hive yet to he found In green tobcco, .nlgktr rites of nitrogen fertllrier eppllcitlon II isiocfeted rlth In increise 01 the voletile end toberco· spcclflc n.nltrolinlnes In the srocessed toCicco end In i level In lurle) ind ~lur.Cur~d tobicco cen double following idded nitrogen fertlllter ippllCitlon 12,31, · Thl ~~tt (orP (ELUICII IIrYCtur)J II1eEltd 10r Ictmpcl j ~~~tlltttng his prabibly no influence on the I.nltro. s1Pine level 131, 35, 38), j · the W.n~lro~~ntne level In Burlc~ tobicro is p~rtleulwt~ high beciuse it i higher nitrogen rcpulrlunt thin Ylrglnli tobicco 128, 311, ,' · Annulltg repelted nelsureaents Eoniltl thlt the Cttwtfr · conditions oreveiting during tobacco cultivotIon Ix~rX Z 1 ne1or influence on the O.nltrol~nlne level, the arising mlr~lun teaperltures IDPI# to be nerticulirlr i 1 Inport~nt (27), P · Sane studies hive reveeled tklt there ire probebly no definite interdependencies between leef porltlon ud nltro~~nin( Itrol In tob~r~o or oott (1, 21, 281, ht : other expertnentl hive shown thit the nltrol~~li level In~rri~e~ with rising leif position (tf·23, 621, .fhl tlne Or the dlf It which Burlc~ tob~~eo is harvested his no Influence an the titrate content of its stens 4. i: or Iclve~ t 32) , B --- ~ Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 ~4- I *n, ; '21' S,J,l~u~n~ Ihe curing method hlJ 1 n~Jor influence an the nlttosinlne concentration In ~ob~eco end Elglreltl I~ote (3g)', fhe high teaperiture associated with tlue·Eurlng (up to 90'e) can till those aier0·0tprsllnl thlt trlnlfore the I~tilOldl fnto the tob~ceo·rpccfffe W.nltror~mfnei an, tonsel puentely, the I·nlttol~n~lne level in Ilu~·Eurcd toblcco~ L is eiclrt~ lower then th~t of ~lt·curcd tobaccos (28), · the climatic conditions previftfng during curing c#rt e Rijar influence on O·nltros~~lne formation, 0,9, the relitlve humidltr during ~lr.cutlnp cin he of Inpott~nec (Zf), 1 5tllt curing results In higher tobicco·!pc~lfle I·nltro· Sin~ne levels in the smoke than orimed.leef curing (27), · jun·curcd OrlEnt tob~Ecot have ~lrtCd~~ lower ~·nltrO. slnlne levels then flue- end Ilr·eurcd tobiccos (fO,t3), · oosslbly, the curing method may influence the rtl~tlvi COn~entratfOn levels of toblceo·rpcelf~~ O·nltto~vlna~ clna, Iwn, lu, nne, in relltlan to etch other ( 10,131, · Ibe reduction of nitrete to nftrfte th~t t~tn pl~ce during curing (b~ctctlologle~t Indlor cn,m~tlc) Influ· ences the forRltfPn of I·oltrolln~ner tron precursors (la), · Ilr·cu~lnq enperlacnts conducted on Burier tobacco hive Shown thit eonrldct~bly higher levels of I·nltt0tLlneJ ore formed It 1 curing taparitura of ]Z'E then at 16'C, it is ~tro ~ir~cfittd with e rise of the nttrlte level in the tob~eco (~9), · Acc~lcr~ted curing of crude tab~eeor (81e, Comogenfied leaf curing) (4,11,61) limits the bictcriil influence, the I·nittaiwinC level In HLe hoaopen(2ed tOblcCO Ielf is iignlfif~ntly lower then thit of Ift· end Itui·tutld tobi~~or (21965,831, Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 ·i t~· 3,3,3 Fernentation · lhe partially anaeroblc processes taking place dur(np fernentatlon pronote the fornatioo of tobicea·ipceftte k·8ltr0llafl(l froi the tobacco Iltl~Old~ (22)1 · increased nitrite levels can arise p~rtferl~rl~ In snuff an account of ov~r·fcrae4tlt(an, and these oronlote the for~~tion of I·nltrorintn~i I17), $" 3,3,l'foblfco eonstltuent fubttl~ .The bacterial content of tobacco is of decisive Inportinct i far reducing nitrate to nltrite, and for the nftrosatloo of ~l~ilofbr (8,17)1 turtherr;ore, alero·otgiRI~PI can produce substances and rnqaer that :an catalyte a direct ~j2Q ultroiitlon reaction (a0), The reduction of the I·nl~ro· samine level in snuff In recent years has prob~bl~ been achieved by naintaininq 1 better control over the bacterial content In these produCtl (8,61)( Iftrlte farmatton II the Ilittlng factor far the creation of tob~ceo·fDceltle I·nlt~0llitnel In tobacco (c2,19), The nftrlie level Of Ylrglnfa tob~ceo, which contains conparaLlvely low puantltles Of h.nitrosanines, II clearly lower than In other tobacco tyoes (tB)I .the nitrate level in tobacco is an inportant factor for I.nltroiipine forn~tfoi In tobacco (j0,33,43,651 and in Joo~e (21,23,29,31,(2,a31~ It is necessary to note in this context that the nitrate level in tobacco blends is oarticularl). influenced by its share Of stens (1J), But it rel 1110 cltlbli~hed that the nllr~te level da(i not influence the ~no~n: of NHc foried in the uote [76), ~a Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 a b '" C · I doubllnp Of the nltallnl Itvll In tobirfD trobtll tll ZHH rlclO In n~fnitteln ~not~, In I·lold Inr(e~n~nl of the nOinlcOtln, IOril Only tiiultr In I la·X rlii Of III, Since the nlCOtlnllnOrnltOtlnl rltlO In tobIceo It Ip(rOi, 15·20 : 1, nlco(tnr iuit bi rcpIrdid II thi prlnclpit pracur~ar for IIn Ind niII (2,36,(2,63), SOni eipirintnt~ ~rviilid thIt thiri II prObibly no corrilitlan birrcon thi nteOtlne Ind nornlco~lne IivOt In IhO toClteo Ind the IIM levdl · Dlverre IrIf Ind !no~, cJnitltv~nt IUbltlntlI Cln e~t~yle nltr,llll~r IDln~llon (Ihltcllnltcl, ~~rhlnyl e0np~und~ Ite,) or ktr) (n Inhlbttlng,lllot t~lcoholl, EllbOkldrltll, lid Othlrl) (10)1 ' i r · Studtrr tndlcite I nigItlvt corrrlit(on bitwctn the luglr epnt~lnld In talncea Inn tne I~L Iorll In I~ln· ItteIm Iio~t (21)1 II thO iupir IOvtt rliit whltn tht tobIcco nlttitr llvll Ilnult~nioully dlElln(l, then thi h rtlItive rItlo of tob~cto·lglcllle ll.nItrntInlnet It luDDoted tO chInpc: Ilf Ind One levttt Iri hl9hir thin Me wtl ~n~~~nt~n (~I~ fi .bdtanrPlr tr pnln~ ~ d~l p in ~8u~r for tht lori~tf0n Of 1511 In tObIerO Dtaduets (55), .the tolleeo'l pH·v~lul hit no Inllucnce In M.nltroll· Ilne forniffon In thn IiIf (15,17,651, . ~ ~·hour elttlctlnn of ehewfno toble~o In ~~lr~ d Ciulr~ I 10·I rile 6( NII (65), B t d ClibPDF - v~~~~io.soni ·26. 1,3,5 ~E~9, l·ye~r 1g1ng Inrreared the 512 level In ~(r.curtd burier ~ob~ceo by 240 1, In nit tobacco fron OS to 1121, Ihe role of In~reat vol higher I~ be 19119 teooeretore or Jole than it ZO'e (1), 3,316 Sioreic · Depending upon storage tclptrlturt and durltloft, the level of H·nftror~alnir in snuff salDnles rises when they are stored In Closed p~~tfnp~ (8), · Snuff stored Il open DLc~lngl is I~lOtlltCd with i I·lold higher rate of increase of t;r volatile I·~ltro. ~~nlnci over closed pacL,inOs (cootvlnitlon with b~ctcrl~ 1) (6), Ourln0 i 2·wllt rtprlp( period the tSII level rose by 50 X 656,66), · The III level In ~lr·turrd tob~reo role by Js I dur(nb l·yr~r storage it 20'0, as opposed to to I it JO'e, Pirillel to this, the nftrite conConttatlon declined wore it h~phr~ tenPerltyrel then It lower oIes (12)1 ].].i Iradlrtipa i .rAC relatlvelr high nltrete level in stews ~y pairrbl~ influence the fornation of W·nltro~vtnCI during ~ tobacco processfnp (63), .He~t treitn~nt of Yfr95nii tobaceo was associeted with a 50·1 nllror~Rlnr reduction, as opposed to i 100·1 increase with 5urtey tob~cro, The rise of n.nitrosiwlnes during Droduction averaged 31 Z, while II stens It rose by jOO 1 (39), Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 ~------ I' 'r;~ L t · tt . B B 3.3,8 IddltivCI Three nonlhl efter ~l~.lurlb tOblCCO hld been Ipriyed llith n 10·t IIcorblt LEld solution (82) no reduction of ual~!(l~ end toblCtO·IPltlf~C I.n(troslP1ne1 In the ~i a I~otr could be detected (JI), The tene results vls obtilned when the tob~rro stens were treeted with ~n i`~3i Iicorblc Irld solution (II), · liplrctte lo~~ceo sprly9d rlth t,Sd snl 10·1 Olucolr, IUIIIII 111 111111 IYI11 )(lllltl 111 11111111) tlll~ll: Glu~oic rlpntfi~~ntly reduced the tob~cro·~piclfle N.nittoJlalnCI, p~rtleul~rll ICI, In ~~lnJtreY Ir0tr, Reduction nls less distinct when 1IYCtt SYglr wls used, f' a whill klW declined only nrgll9lClt with sultose . ereA less then r(th tnvert lugv treltllent (7511 ~6; .rhr toble~o·rpce~r!e I.nitrol~alncr In lob~rco rautd be reduced by eddinp sugir to flue·curcd tobacco, the level wel decreased still further by ~ddltlon~l tob~cco he~tlng (30), ii Is sulple(ed that I lorpholfnle ossfn9 Intrrf9t incrlll9l tne n.nftr0l0lorDh0tlne lerll in snuli ii), 3,319 Pic~lnp . Indirilunl cir·tlght p1(k1"9 of snuff In ~lualn~ua foil prev,nts ~dCltlonal lara~tlan of 1Sllh in the stuff partiont (rpnt~afn~tlrl by RlcrobtOlnlla 1~ elludad 1) (9), .lh( lorpnoline Partly rontllnrd in thn p~etlng nltcrlul for Snuff con bring about 1 diffusion in the toblECJ r~Crt it is nitr0iittd (71, ru LII Clit; PDF -~~I::! . f3StlC, i: Olil -- 9 I, ProsDrEts ~nt lollowlng study Pro9ri00cr could holo to coin laund tnorlcdpl on the IOr~~t10n (nl reduction of h.nl!i0linln(l in tobutto prOdUCtl, inprarcncnl of Z.nltr0iinlne lnIiylIts to doll wore efficiently with the tirPe nunber of IInlolo'thIt Ire repu;red Is 1 result of the ~npl~i tobIcto ~~trlr Ind the colnoliceted Intcrd~pcndrnclcl, Rlnp tests conducted between fndlvldu~l Inilytirdl I~boritorlci could halo to P ~~llbr~llon stlndlrds), dvo/d enIlntnItlon errors (e,9, ertCilCt fOr~ltfOt, lnEDrreCt r ..) Ynrllicltlnn 01 th~ Curlnp Il~hnlgu!!, pct~leul~rl) t~~ influence of n~r9ln~l conditions such II drlfng tfne, P I~npirilurr, r,lillri hunldltl, lelrli of oirlcn, · It II i neCelllrr to clirlfl why the l(.nitroIIninC level is so r conrpleu0ul)r low In ~un.~urcd Orient tob~rco~, · it epteerl to de ldvintigeous to renloVe the stews frOn the 1/i1nl 1 during the cwIno ptocclr, · t!ltr IhOuld bl rolduc~ll to ettIblisl. whether l.nlfrq10p~011na is IultIble II In S (ndlcltar subrtInei for the forPItlon of I.nltroi~~lncr a In tob~rco products, d ..) Use Ind development 01 Ielsurlng nethods to Ehect Ind control Ihl b~ctcri~l conlnnt nnd cnilme IctlrltI In I P toS1Cro oroducts, Josllbly, ICLIUrl~ cIn he tIl(en thIt d Inhitlt Or even prersnt IlcrobfIl or Inim~lle nltr~te e ridurllan (c,l, luring le~itnt~lllnl (~ddltlv~i, rIlIILlgn, exclusion 01 elrl pissing, ~lr~ol), .,, ~ ~~in~lton of groduetlan steDI Lhit could influence the a · I·~llr0llninl levrl ourlnp tobiceo ptilirillon (tT, SrS, C al, rl~plia~ ~*~( kd, rK~1~J nrOduCll, HC ~~s~~~l ~*xul re d~nnl u·~b·l ~UMI I 01 luring, tlmlc'itu'll tinc, nolltur(, blind roipoiltl~n, a ett.)~ N CrJ ClibPDF - v~~fa~io.soni ·20· '1' ··) Iltlllilhi~9 Lhe (nl(~dCDlndCOC1CI Lctrern ~I~ Irrrl (nltive Or I~ Iddltlrl) Ind the I·nltr0~lnln~ Ioril in tOblCCO lnd riotl (type of iug~r, ron~,otritlgn, ~tch~nll~), ..) Ylril(EltlQn of the ItOrlgl Eondltloni, both of crudi tob~ceo (eglng) OI rell II Of the flniihod produotr (Cf~!r(ttl~i find cut, ctr,), lne lottowing Influlnrlng virl~bl~i could de ril~v~nt for M·nlttoIlnlno foti~tlon; O hunldltr, Ilr ~tcClr, pn·vlluc, nlcto.orglnfln~, Intllll, ~tor~pe Ilne, tclplr~tur~, tob~cco lot~tur~, Invlron~~ntll h c~t~lytle~ltr lctlv( eonltltucnt LubltllCCI 01 toblcco, p~Ctlnp ~~fltl~l, typa of picllnOl .., Irr~d~tlon tciti to dlnlnl~h the k.nltroi~~lni Iirrl In crude tobiccoi end Ifnlrhcd productl (poiilLtl In conbln~tlon with edlltlver to collect the Ilbir~t~d 10,), ..) Oeternfnetion 01 the re)itluc M.nftr0ll~fnl Icvel~, Polllblr, conclullonr C~n he driwn fronl the raletlre concentritlon condltlanr between IIM, nlI end IAl for i the origin of the t0blceol end for procirilng I,lrurir, CII~P~F - vww~~io.coni ' y i "30' 51 I11ECLTUk II hdP~IC~,k,$lnJ,I nbcrblur;W~R~BWtDnlJ,LCwltOntE,E,Y0drr: ehinOOe In thcnlol CcaporltlPn o( ~o~wml:d 1JSwrd )nd I\lrlCyrd klly IObKLO S~wd In tontrollep Enrlrsn*II;J,AP~lr,ioOd OIMt3~3~bbf·WBI 1198;J :I Dlr~arl~ln,Y,J, Ik,F, kvwtonln,61Strphn~cnI ~*I, Of ~n~lro~cncmlrotlm In foa#ep: kpn Under Vlrylnp lbOnOnlC epldltlOn~l fOb,QCI~ 1Sb·IZ IlsJ1 ;1 HKht,S,1::,8,5mlnlOEnalR,n,OlMitD,rb~funnlrlC,rlol ChKIEII Studln On Tob,ccP k~r~LI:Studln On Non·Votdll~ Llrolwlnn In TnOlCEOr iln~tr,t~b~· 51 forreh, Int, e(ll, ldl(lDnl 11 ~~o,t,C,:R,lorclD,w,DFJonP~ Ilowo~nr~~d Lu! twlnp,l,th~cn~aiul kill k~d Sw Rcl lun~rv R,d ttl A bl T~b~Cl#Xh~ I)IU·Il,l IDfSl fl br~nnrwnn,~,l, ISIJllnlSIEIQWIOIHoillmnl OEEurDnED 01 Iwtltrc~vina In Chewlnp Tob#~olC Glow Iw~l2th toa#ea eh~cldllR,arCh ~o~(smC,1 F~tlrr~~~,tb~, 1984 61 Frunn,~~nn,T,DI ~Dlwo~iwni esrurrwlt 599 Ilrw~wr~Phlr Dctmlndlon 01 U,tllr,Nonvol~tile ihd lob,ECO 5OCCI(L[ N.YLtrO~llln~l~th to~weo Qlnll~' R(u,reh Conia~na,Yluton·bI~,GI,198i 11 prvnnrHnn,T,D, ;J,e,~FottlDIHolivn~I H-NI~r010~01ph0111 Uld OlhOT Vdlltll( P BI Owtrd~hl,l,·C,t,LSIwKhl Voi,tll, CltrPH*nn In Snul(lhd eh~l~ j H-nllPljnln(l In mulf ~Db#tDIC1~LlnWrmlll Ib)~b~Jl)LI(1~Bi) L TOOKED On The 51190190 ~UtlflFd101~~,10x1elM ~~FIOc11985) or HOc(nlnn D,:J,D,lld,ar: e~rllnoomle tob#Eo~p~clilr N-nftrorvlnn In Snuff ,nP In the %llw Of Snuff DIPPs~lEr~w R~,t~l~O~#, 119811 101 Hot(nunn 0,15,5.HKht:R,n,01#flEILYvndr~ H'~Nltrolen~omleotlnlr In tobKrol ' j SEIDnEe IBf, :15·;b~l(t9f~) IIl Yoodl 199, v, 5, IS,J,LnlJ,f,e~~pl Inlwl slkUot~:huat~l~,s~mla~ DcProteln~:~tlon EflKt~ cn nan0Q~i:~d Cnl OurDd IKCI DrOduEtOI Prl tr, t,b,l irrlch, Lt, I: I4lltll·:tflll~B~I I:I not~wnn 0, IJ,D,kd~nsl:,OIPrunnCI1BnlS,S1HKhtl FOrYllb~~GCVt~nCl~~nd I~Cln00CnlEltr O( O·lltrDY~lnll In tD~KtD ROdUttll~i~·lOmIS#lb~PL Srr 1 I~J~:9:~'t; (loB1, I:~ ~lul,w,:n.luhnl On~sluchuacn ueOer die IreMfludtlpm Imltra*lne 9* ~~:,I·,ll~lOldr: Flrh, nI tt,~n~s .t~b3,lb~~Ot~l~ l1'lrJl 5t III 'I~~~ncnn F.:J,D.Sd~nl:):.O, Frunn~n~nn IS, S, ukrht: IhW~nM: 01 fOb001DKI(I~ u~:truilml In to~,c:a kodurtlle~nr,kn,39,=5~5~.~'~~ 11979) 151 Ir~C!,~.S, Ifi,n,orn,~:DI~~ iunnl Detanin~~lan Of H'·Nltro~onornieotinr In Tc!iceo bl IllOh SpItd 1l9vld ehroHteer~Dhv:~n~l ,Dlolll ll:)~?Pll·~18, Ile:fi Ib~ ~~4t,5,51;C~P15'lm: 111Hlrpt~lk,~, Qn~~:I~E, ra: D, He~iunnr ToD,Eea.SPK)f11 C; :'~Ii·~'~l!:~O~m(tl 3~ Irga YICttls In Vitro end king t0b#Co turlna end ICI:;"3;t.ISlt, In S:r~ln I nit,: J,11Etl.ClnC* In~t~bO I1II B19·El. 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JI I·Rl~o~~~lArr II tob~eD ,al In klaltrl~~ Irl SIdIltrIII SIoII (rr elo~rrllmlCmt.fn~,3!,3~tB·::;:, 11977) 191 H~II~~~,D,~ .D.Cunn~*nn~,S,Ybbl IIl:,b ~1I(11I NltrOlllln)l In I~wro Ad +IPC(rl~l Ad SLdll(llrl ~II Ad IndWT BIIIPUIII(IIIY ~1I~1~~19·Bf~llPB:l 3~ ~run~nm~~ ,DIIDIClln~nnl Ill:lr N~htrPldl~lhnot~nlnl In lcbIrtn Ad e :II Tlo,T,e,l J,~,II~~IDlf,~~lal RnI ArannIr Flctrl AffKtlnp ,DI~L~- ~rl(lI,~~l~lr 01 51~1111 AP IPbIteo Sprll~e Iltru~lan ~~ ~so h~ur~ll btC nnlInp 3 th, I~,~(~rr,~tllm I :;I Holfn,nn 0.:J,D,b~~llb,~,rl~l~19151~hll brlCI1 SludIII 3 ~KCD bit Ifvlll,Arlllll3( L)II111~J TOrtCeap~~llC IltrA)IIII jl Tlwf) ~rw~rtlllWt frl,lubl, NofIl ~f·516,C~~11~83 :II ~w~,~,D, Ir,O,tmn~~ol),Ll~k~nl D1~IE,1 S1udln On TllclC b~l 1I1~ Rlold ~rtho( fo ln, IllIIvllloi t8lrta.l0ll'le ~ltr#~~llr )r (~~11~14 ~brLltQrqll 111111 I thaul a·pr n~lul*1J.~~t~~l~WII IIIJI 3) kv~~r~~~~ ,I,lO,nof(~nl ~l~l~t ol lh, ~~c~arrle kltr0ldl' IlkrnolInIn II t~~rco IrnduCtI Ind !:?:ceo lLOllI ~tl~Wln(lll~ (Ij~ 11111 a loa,rra Of 0,5,1, t~or~t, C~ :71 ~t-C~ot no,ED, ~n:·El cl~~llJ,e,l~l IlIlI Isou:I cllII,l tiiKt Of W~r~~tllO Ad O~ P~YtlC· 3 ea,le,~mr~lr~llk~ &(nlI OcAIIrIIn 3 Lrllr InIKrI :81 Curlno t*bllUrJ~TCI Wltrownll, E·trt I TIICCOII klll*ry Stdr FI IP·P~~I A,LPI rl.l; 11~1;11511184~ nIlI IOPu:I ku:,I liint O( W,r,~ltlnp ~d A1~1~ klcttln 3 Rwlr~l~m~ic~llka 3811 OoIpIIIttIn 01 ~rllr tnIIcco )jl )CT'~tO~t RIFD, oI; IIIB;'IPIDIC~I(I )13~)IJll Lt~tlltlO Of Iltru~lsa In tatIece ;) I ~l·r~lat 8, ZD, I~ tl(O:II DI~IL~I)ID( E~J(I~~Jl~phl flO)IIl t(b~l b ~r~rle Clctolr ~nllrCLII( 11) ~11~*1(I Of LtflC 3138 A( rltrowan IFfli rl'E:IIJ.S.J~rl ~elr,~,~lteh~ll~ IfItrItI ContInt 01 ~~rllu 1~)ErD 11 Bllvll' ;~ ~l·R,se~~ ~,lb ~ iF~I LZC,nl r·~t~olnllsllll In 11*10 ::I ~a·r~lcl: r~,P~,I,:?! 1i3'1111 t,e.O'llPI mi Ef(Kt O( IKDlIIC kid 3 r I~trlr Dldr Ld U~qlr?l~~;h, Rllr~:~n Trr A ~lr~d IIInd B ~L1·~.~5'' I?~~ '!OT:II I~Ple*~r:1~P,nl tch,ll,DIJ,WI UnIIrlIIv 01 ,(~!·~ qrrOL:t 13~7.~~t(l~ll DI Bltr~y~~( CCtlll:IIL 1I1( A( 1911 I~(:~G O ;:· E::·'r~c. ~~',F~ Ilc:El; ~,5,~~~lne~ tl,C~~Spcel(lC NltrWlnlnlllA N F~.'· I W g CII~P~F - vww~~io.coni · La ·32. h ftl F;f.RCrYVI NO~LD,IOI,O (19811: ~IDIChCIPI~~J,DV/I; 1~61111tlhllll tillCt 01 noIvddMl* And H.SH1If r)n nltrorvnr Dcllvny O( burlY loD#co !81 p~r-sPoert Ho,kD, ll:-84 1199111 S,(l~l~ ISOU:I 019,'11 I((Kt 01 NICDtlM 1818 NlIrItr On Nltroluln) F#ll\lm (n Tro Ikrlw lobwso Vll~~ln 591 B~l.kCvlk 119afllD,1I,JJ)fblnt rOIlll#I ~10~ fOblC~O SnIIlfII IfItrnIonIyI Opllwll Fcr R~u~tlOl II)) Irclnlcnl Arpor~ S,lnt,~wllnp On II.HltrPID tP~OPund~l Bmti,~lbrb~kn~d~, Sot,i~B: LP.~tll~l II) F~nlnlm,P,A,I~,;,I IIpOTbEUHI ~0l~.~r~nt Irrdlmt k\d m, kruml~tlen Of NllrItr And N'·Nllrolwwrnltot~~~ In )rlw Icbrrslfllllt Bcl,~ub,SI,BI~·BBJI 119841 I:I nltlolr~e,t,~F,Elu:IIJIIISlmlllYIPl~shl TO~Ke~SP~I(IC H3lltrOHIIII~I E(trct Of Burin llll~lo16 Ilollnr. 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C~-- j ·33· Elk olt,J,t, I),P,trunarnnl O,~llunnl N·llltrorOpr011u,An Indlut# lor 1 [Oll(tr~l(P,kllCIPIIINlr, , O~t,198: H~Hllr01)110n 01 I~lr~n In TDb~CrD hOdult~l;bW 10~Y~O Dmlltl'knl*Eh ~' Ho((rnn, D,~I;,S, nrrh~15,1,~6,91) ,O,Avm*anlR,II vmlenlE,I,Ymdrl 1 bhnl~p,; III~Chr 11·Wltrow~lncl ii ld~lh0,lh~lys,Blo10OLICC IlltlYltrt und StClfwf~hal:\rortrH) bpl Ft,;OlJul1 1181 511 wKbt,5,5,: I,khsl~:;f,~llnnnl tlltrlrnaul eorgm~ndl In Qpwat~ Slo)e Ind their Folslalr kKurnorrlkKmt,lldr,tob.kl ,;,5993, 11011l 1 581 ~llal~lh,H,J, 11.5(jdlsl Unrerluehuneen:ur 8110900 wI nJ(l~ro~lo~~iot~n I·R ~llolln·ll'·l~rldl ~lll~nt~~bll·lll~ 11918) 591 f.:uc ,J,),r thInnr In lhe Cboo~noul Cmltltuntl of Qln To~Keo Dying I Curlno ,n~ AalnOl~ob~Scl,l, 9;·)b, (19611 b3) &~grl~ln,Y,:, Ih,r,llrmmddl An Illt(mltl nCthod for 500 IhIlnll 01 I H·Hlt~Dlcnornl~e~lnr In IOb#COIJIP4rlC,F#j Ulrc~l~*911,11~1 Ail ~eJwlp,~,ii,:J,LIII EILollrlElrOd~lllC~ t101 ~OMpll:~ LHf klnplkltr, tit,l (orrch, Inl,B (;),Tjl~l,(l~B1 ~:, tco,l,C, IJ~~~Ch~PI LnlJ,Dll\dmllD1HOf I~innl 510011 Eort,iltlon nd IClf tlrll j L~O~C~~IWI ,nono LN( Wnd S~Okl th~r,e~nlltla of [kfl~y;abKcollLllr, 1 Tlbd (ol~~h, tnl, II b;~ nollunn,D, It~Rlth~Jlplr,r,L1U1 EhHIICIl ~tudln on tobIoeo woc,llfVl,0n thl 1Jatton ~nd Id~nt~flectlon 01 Mlftlte JnP non·v0lltllf naltrowlnn u~d hrGI~:ln# In EtPVlttl YOLII11~ SC~I~UblPI1JFlbSI (19711 bJI OrJtrrdAl ,B,G,:5,SI#whl N·#llr~inn In vnrlt Ind brirg t~reo m lho S~d~~h vrl,t In 19~51Fd,l#dlt,e~t~r 1(1),299~, 11980 bl I Hol im~nn,D,:S, 5,HKht:R,n,~nd tE,I, YyndHltle, teel Plalol Stud~ n L fob,cro Snell.llll,NI trolonoroeatlnlknma In tobreo,Rutim And elrFlcWMIElty:IPbC kl,~U1~1~1~~~07·~01 11976) i b~l nec~n#n,D,II',D,8~unlcynnlj,D,lkl,nl5,SI~chtl Forwtlen And Ihrlvl~l 01 #·Nllro~En~nc~ In 'foblrco RPdUCtl And their Endopenouo rcrutlon In C~nan*l; lAkt k~.hb1,57,113·76~, 119801 bit Hg!(nynn,3, :n~DOrOlglS~~Kht: b~lpln In tobKto kcr d w'ltrolonor· nlcotlnPlr tOCCeCe.5GKl(lf tlr~lnDPlt ~1~ eOWnlC)tiO~~lJ,Hlt~ln~n In~t~lP, 1811.1511,119771 aPI Sr~n~;t;,l,:I1;9~d~ :D,Hgilc;t~l knoroOMIC Po0mtl In Lhbwnrd Pr#lluQ IOIILCCI:#.1I tr010dl ~~hlnll)nl# And 1( I-Dlmlhrt hrdr~:lnlt~nC,~dt,:, I;Z1X,fro7:I 1;: Flr,~lP,F,: FE·Flp)l Ilr~ 1197911019 BCltllUnO r~ mlItrolInlnen In tl?~r~t~C"rfCb nll dr thHn~l b~lr0v LhS)V;V IfEA) i.~l flr!cr,F,: F5.keDert R~,:l 1195:):019 kltlnnulp vOn tlb~~lSClilll)m K.lll!rSli~:nPr~ In l~bll und In RWEhlEndp#t ceC·fEWopplung), 'II Flr·!tF~~F',: FE'U~lc~lr~hv~O vOn )~UtED)LM deI dhltS~hM ~~)fH (I~kll ::I F!rl)rlr~: CEVl(et~CC'I~M~ ton khnvp(tlb~l~n dll dhltlChn IIrlln llIell ", I:n.!r,,P,: eE.lh~tCr!:I'.~Y~O ve" ell~rl!~(ntlt·)len dr dnt~bre Llltrr ~]O~:' :: F:~,\~,r~~,! 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Re. ~, 935·911, 11985) ' i 81) a~ror, Q! ~ultuplfn t, lo co~toar 19aSI, n·il~ 821 nlrrtrh, 5,5,, L, Yeltcave, n, Ben, , Shublt: karb~ta· ·· of bfclnopen~r, k.lttr0lo·Coapaundl, kbw 111, 6547 I: . nitrlte Reaction: Po#lbll Aml of BloeUng the Fonetion 119/2) ·, j 83) nuaon, J~w,, I, Abdlne: Dcl~nloltlo" nf I.nltto~onornirotlrw In Tobdcto by e#chrom4tograDhlrP1II 5Occtrorcopy, kill I let, 10 (10), 117·786 (19771 84] B~unnemann, ~,D,, i, Cenoble, O, Holfrnnn: I.l(ftrorlalnlln Gnewing Tobeccor: An Internatlon~l ~o~p~rl~onl J, Igrlc, 1 Fooa Oleml 33, 1118·1181 (1985) 85) Bharddr~j, v,P,, 5, Tl~yama T, runade, A, Tenilnure: 1 n.nitroSon0mieotine585·586 (1915) In J~pc;ne Tobrco Rodue:l, bnn 61, ,O I RM- JI· Inner P O N C I I b pQ ~~?M~-Lf~ st I orze H~ i "1 ~ .]~· i The FE·ll~er~l~re and patent recherche on the subject of h.nitro~lmlnc~ In tobacco products revealed the following Fefzrenc!i in the lilted data benti: Data b3nk Wulaber of references Cj 1 II, Internal, i,e, Interbat (1 7 Stab (Scil!ntific and Technical 139 Alerting Bulletin) Patents Z 188 13 i. 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