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35th Tobacco Chemists' Research Conference 811006 - 811009 Hyatt House Winston-Salem, North Carolina Part I of II

Date: 06 Oct 1981 (est.)
Length: 43 pages
04307303-04307345
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Area
LIBRARY/SUBJECT BOXES
Type
PAMP, PAMPHLET
AGEN, AGENDA
LIST, LIST
SCRT, SCIENTIFIC REPORT
Alias
04307303/04307345
Site
G39
Request
R1-004
R1-060
R1-061
R1-132
Named Person
Abel, S.G.
Arcila, J.
Arndt, R.
Arrendale, R.F.
Baker, J.L.
Blume, D.E.
Bohlander, P.J.
Borgerding, M.F.
Browne, C.L.
Brummer, U.
Brunnemann, K.D.
Bush, L.P.
Cartwright, W.
Chamberlain, W.J.
Chortyk, O.T.
Collins, G.B.
Cornell, A.
Davis, D.L.
Dirks, V.A.
Dube, M.F.
Dwyer, R.W.
Einolf, W.N.
Elliott, J.M.
Enzell, C.R.
Ferguson, R.N.
Fleming, M.L.
Glock, E.
Gordon, B.M.
Greene, G.H.
Griest, W.H.
Gwynn, G.R.
Hamilton, J.
Heckman, R.A.
Heeman, V.
Higgins, C.E.
Hoffmann, D.
Hyde, R.A.
Johnston, W.H.
Jones, S.T.
Katz, T.
Keith, C.H.
Kinser, R.D.
Kung, S.D.
Lam, J.J.
Leete, E.
Leggett, E.
Legg, P.D.
Litzinger, E.F.
Long, R.C.
Lynm, D.
Marmor, R.S.
Massie, I.E.
Mathis, D.E.
Mcfuffie, K.L.
Mcmurtrie, A.
Mcrae, D.D.
Mohpatra, S.C.
Nishida, T.
Nishikata, Y.
Noftle, R.E.
Nordfors, K.
Olender, V.
Olerich, G.
Pandeya, R.S.
Perfetti, T.A.
Pitner, T.P.
Poushinsky, G.
Reynolds, L.
Rhodes, P.R.
Rivers, J.M.
Roberts, D.L.
Rosene, C.J.
Samejima, T.
Saunders, J.A.
Scales, J.R.
Severson, R.F.
Shenn, S.J.
Sims, J.
Slaven, R.A.
Smith, L.B.
Snook, M.E.
Song, B.H.
Spears, A.W.
Story, L.D.
Tso, T.C.
Wahlberg, I.
Wallin, I.
Weatherly, C.H.
Weeks, W.W.
Weybrew, J.A.
Woodlief, W.G.
Wu, D.T.
Zilkey, B.F.
Document File
04307303/04307817/1981 Tcrc
Date Loaded
05 Jun 1998
Named Organization
Ahf, American Health Foundation
Amer, American Tobacco
Analytical Methods Comm
Bat Cigaretten Fabriken
Boehringer Ingelheim
Bw, Brown & Williamson
Celanese Fibers
Central Experimemental Farm
Central Research Inst
Consolidated Cigar
Coresta, Coresta
C H Dexter Division
Eastman Chemicals Division
Eastman Kodak
Japan Tobacco & Salt Public
Korea Ginseng & Tobacco Research in
Local Arrangements Comm
NC State Univ
Oakridge Natl Lab
Pee Dee Experiment Station
PM, Philip Morris
Policy Comm
Program Editorial Comm
RJR, R.J.Reynolds
Sea Ar Beltsville Md
Sea Ar Oford NC
Swedish Tobacco
Tn Eastman
Tobacco Research Lab
Tobacco Safety Research Unit
Tobacco Science
Tobacco & Health Research Inst
Univ of Ky
Univ of Md
Univ of Mn
Usda, U.S. Dept of Agriculture
US Tobacco
Wake Forest Univ
Ac Monk
Agriculture Canada
Litigation
Stmn/Produced
Author (Organization)
Tobacco Chemists
Tobacco Chemists Research Conferenc
Characteristic
MARG, MARGINALIA
OVER, OVER SIZE DOCUMENT
UNCO, UNCODED LIST
Master ID
04307303/7345

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HOSTS WAKE FOREST UNIVERSITY R. J. REYNOLDS TOBACCO COMPANY 0 O A W .i O »'~
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GENERAL PROGRAM TUESDAY, OCTOBER 6, 1981 3:00-9:00 P.M Registration 8:00 P.M. Analytical Methods Committee Meeting WEDNESDAY, OCTOBER 7, 1981 8:00,A.M. 5:00 P.M. Registration 8:1b A.M. Opening Remarks and Weldome :30A.M. R. E. Noftle, Conference Chairman, James R. Scales,, President,, Wake Forest University SYMPOSIUM: Regency 1, 2, 3, and 4. TOBACCO LEAF CHEMISTRY: ITS ORIGIN, UNDERSTANDING, AND CURRENT TRENDS Lance Reynolds, Brown and Williamson Tobacco Corporation„Symposium Chairman 12:15 P.M. LUNCH 2:00 P.M. Technical Sessions: A, Regency 1 and'2; B, Regency 3 and 4 Session A: J. M. Elliot Agriculture Canada Presiding Session B: C. L. Browne Celanese Fibers Company Presiding THURSDAY, OCTOBER 8, 1981 8:00 A.M.-5:00 P.M. Registratibn 8:40 A.M. Technical Sessions Session A: D. E. Blume United States Department of Agriculture Presiding. Session, B: C. J. R'osene Brown and Williamson Tobacco Corporatibn Presiding I
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12:10 P. M. L UNCH 1:40 P.M. Technical Sessions SessionIA: L. D. Story United~ Stat'es Tobacco Company Presiding Session B: E. Glock American Tobacco Company Presiding 4:30 P.M. BUSINESS MEETING' 6:30 P.M. SOCIAL HOUR: Garden Terrace and Reception Deck Compiiment's of R.J. Reynolds Tobacco Company 7:30 P.M. BANQUET: Regency 1;2,3;, and 4 Philip Morris Award for Distinguished Achievement in Tobacco Science Entertainment: The Auctioneers FRIDAY, OCTOBER 9, 1981 8:45 P.M. Technical Session: Regency 1,2,3, and 4 D. L. Roberts R.J. Reynolds Tobacco Company Presiding 10:55 A.M. CONFERENCE ADJOURNMENT GUESTS' PROGRAM. WEDNESDAY, OCTOBER 7, 1981 9:00 A.M. Hospitality Suite: Room 331 10:00'A.M. Visit/Luncheon, Reynolda House and Gardens. (transportation provided). THURSDAY, OCTOBER 8, 1981 9:00 A.M. Hospitality Suite: Room 331. 10:00 A.M. Tour/Visit, Old'Salem (transportation provided). II
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35th Tobacco Chemists' Research Conference October 7-9, 1981 Winston-Salem, North Carolina YEDNESDAY' MOR.`IING, OCTOBER 7, 1981 Nyatt HdWse Regencry1,, 2, 3 and 4 8:15 Opening,Remarka.and'~Yelcome: R. E..Noffle, Oonferenta Chairmen. 8.:25' SymppsiumIntrcod~uocion: TOBACCO LEAF CHE74ISTRY': ITS ORICIN. UNDEASTANDING~.AND CURRENT TRENDS. LaaceReymolda, Brovnand Y111famson. Tohs¢coCarPoration, Lou14v111e,.KY402328.:30/1 TOBACCO L!EAF:'."A LOOK.AT.HISTORY". I..E. Ndd9ie,.College ofAgribulture, U'nlveralty.of Kencucky„ Lexingcon., KY 40506 9:00: #2 CHENICAL Al7D'PHYSICAL.CRITERIA FOR TOBACCOLEAPOFMODERN'.DAY' CIGARETTES. A'. Y. Speru'e and S. T.,Jones,. Lorlllkrd, 2525.E. Marker Street,..Greensboro, NC27401 9:30. #3 MAJOR'.CHENICAL.CHANGES. DURING SENESCiENCE. AND CLRING OFTDBACCO LEAF. R,. C. Lvngandl. A.,Yeybrev,.Crop Scienxe Dep;ar2menc,.Norch.Caznlina State UnlYerelcy, Raleigh, NC 27650 10.:00INTERMISSION' 10:30 /4 METABOLISM AND'REGULATION'.OF ALKALOIDS'. IN.TOBACCO. L. P. 8+.eh, DepartmenUof Agronamy, Udlverslty of Kencocky, KY 40546 11I:00 g5THE ROLE.OF TOBACCO.LE:APPRECVRISORS IN CIGARETTE FLAVOR. R.,A. N'eck+n, M. P:.Dube, D. Lynm, and J. N. R14era, R. J. Reynolds Tobacco Company, Reseerch, and Developmen[ Departmenr~, Yinacom-Salem. NO 2110211I:30 /6TOBACCO LEAF PROTEIN: MOLECIJLAA.BIOLOGY.AND.GENETIC MANIPULATION. S.,D. XV.ng and T. 0. Tso,.Department of B'Lologibal Sctencea., Umlversltyof Maryland, Balcimore Councy, Caconaville,.hm. 21228 and Tob'~aeco. Laboracory, B'eltsv1~11e Agrlcultural Research Center, SEA/AR;. BeleavilRe, t~ID20705. 12:00 SYMPOSIUM SUlWARY' AND FUTllRE'PROJECTIONS.. Lnte Reynolde, Bravn and Yilliamson; Louiavilde, gY4023E VEDNESDAY AFTERNOON, 0ctober 7,.1981 Sesaion A Hyatt.House Regeocy, 1!snd!2'- J. M: Elliot, Presiding 2:00 /7' THE CULTURAL MANAGET(ENT..OF'FLUE-CllRED TOBACCO STEAM-VOLATILE CONSTI7UENTS. 4. J' A'eske and J. A...Yeybrev,..NO.rth Carollna State University; Ralligh, NC'27650 2:20 18' BIOMAS.S ANOCHEMICALCOMPOSITION OF TOBACCO PLAIiTS.UNDER HIGH DENSITY GROWTH. S. J. Sheen, Deparcmene of 2:40~ P9. P1anc Pa[holbgy, Unlveraity ofKentucky., LexSngcon, KY 40546'. CONTI:7UOUSFLOYSYSTEM.FOR THE.EIITRACTIDBI OFPROTEIN. FROM:.TOBACCO LEAVES. D_ E. Blums, Y: G. Yoodllefand J. J. I~m,. USDA, SEA/AR, Tob'ecca Research. Laboratory. Oxford,NC 27565. 3:00 #10 UPTAKEANDINCDRPORATION'.OP 3N-LEUCINE BY GERMINATING TOBACCO SEEDS..J: Arci7e, S. C. Mohapacrs and Y.. H:.Johnscom, DeDartmenca ofBotany and glo. & Agr. Eng., NCSU,..Raleigh , NC 27650. 3:20INTERMISSION. 3:50 #11 COMPARISON'OP THEDEVELOPMENT~OP MAJOR LEAF COf@ONENTSIN NORMAL! AND A'BUD- YORN RESISTANT TOBAC00. R:. P. $everson,. O:T~ ChortYk, N. E: Snook;. K. L. MoDUffie,. andi. B: S~m1ch„ Tobaoco Safety Research nnit, U!SDA, SEA/AR, Achens, GA.30613 a ._ R. Gvynn, TobaecoReaearch. Leboracvry„ USOA', SEAIAR, Oxfbrd, NC 27565 4:10 F12DIALLEL GENETIC ANAI.YSIS OF'. LEAP ANDSMOKECNAfUCTERISTICS IN FLUE-CURm TOBACCO.. R.. S. Pandeya„ B. F'. 2ilkay;P. 0. Box: 186, De1h1, Ontario, Canada N4B 2.9, V: A. Dirke, H.rroe, A.tarlo, Canada NOR'1G0~.and'G:,Pouehlnaky, Central Experimencal Farm, Oc2ava, Ontarlo„ Canada K1A OC6 4:30 /13 PARTICULATEMAT4ER.VARIABILITY IN BURLEYTOBACCO' . D. L..D¢Vi" . O.,B. Collins aad P. D. Legg. Tobacco and Health Research Instit,ce and Deparc- entaf Agromomy, Unlveraityof! Kencu¢ky, Lexington,.KY 40506 4:50/14 MOISTURE SORPTdON PHENURtBNAINCURED TOgACCO. T. Same]Ima and Y. N:ehikata,.. Cmcral' Reaearch Iuacituce„ Th. Japen Tob'acco amd5alc Pub11c Corporaciom, 6-2 Umegaa~ka, MSBOrl-ku. YokohaY,.. 227 Japan Sesaion. B Hyace House Regency 3'.and4' - C. Ls Brovne, Presiding. 2:00 t15COMPONENT ANALYS.IS' OF CIGARETffE VARIABLITY:D. E. Ma[hie:Tennessea Fasrman Campany, Research Laboracorles,.Eascman Chemieals Dlvision, Eascman Kodak Company, KSngepuri,. TN'. 37662 2:20 F16 REDUCEDEFFICIENCY' VENTIIATED. FILTERS PROV'IDI NO LOWERED CARBON MONOXIDE PER.UNIT TAR DELIVERED.. P. J: Bohlander„ Celaneee Fibers. Campany, P. 0. Box 32414', Charlocce „ NC 28232 2:40 117' CIGARESIE.PAP'.ER.EFPECTSON TAR/NICOTINE.AND CO/TAR'RATIOS'. A. PfcFAtrtrie;. E. F. L1[ringer, and D. 'P. 4u, grovn and Wllliameon Toba<co Corporacfon„ 1600 Y. H11R 5'creet„ Loulsville, KY 40201 3:00 /18 THE EFFECT OF CIGARETTE PAPER POROSITY' ANID PREPERFORATED TIPPING PAPER.NOLE SIEE.ON CIGARETTEVENTIILITION A.VD SMOKEDELIVERY. .R: A. Ryde,.CelaneseFibers Company.P• 0. Box. 32414 Charlbtte, NC 28232 3:20 INTERMISSION 3:50 #19 THE EFFECTS OF VENTILATIONAND PRESSURE DROP ON FILTRATION EFFICIENCY. R. Y. .7vyen 5. 0..Abbi, andM. L..Fleming„ Philip Morris U. S.,A'., Research,Cencer, P. 0. Boa 26583 Richmond., VA 23261. 4':10 !20 A POLYDISPERSE AEROSOL MODEL FOR CIGARETTE SMOKEFILTRATION. C, D. .NeRae, Philip Morris U!. S. A'.,,Research Center, P,. 0..Box. 26583, Richmond„ VA 23261 4':30~ /21A.PORE SIZE lffASURE1ffNT~TECHNIOUEFOR TH'E CHARACTERIEATION OF CIGARETTE COMPONENTS. P: E. Ha[hieand C. H. Yeatherly, Tennesaee Eascman Campany,..Researoh Laboracriee„ Eastman Chemitalb Division, Eascman Kodak Company., Klnigsporc, TN,37662 4:50i /22 HIGH SURFACE.AAPA CIGARETTE FILTERS'. C. N...Y¢ith, Celkne.. Fibere Comp!any, P. 0. Box 32414, Charlbcte, NO 2823P III 0 0
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THL'RSDAYNORNINC', Occob'er 8, 19g1 Sesslon,A Hyatt Houae Regemcy.1 amd 2- D. E.. Blome; P[eslding 8:40'. e2] ALKALOID DEVELOPMENT IN'.NORNAL AND CONV'ERTER TOBACCOS. R. F. Arr¢ndaZe, R. F. Sevcrsenj aM'0. T: Chortyk, Tobacco~Safety'Research.Unit,.USDA, SEA7AR:, Athena, GA 30613, and G. R. Gvym,TObacco.Research Laboratory, USDA, SEA/AA, Oxfotd, NC 17565 9': 00: #24 ~. N"-ISOPROPY'LNORNICOTINE:', ITS FORMATION FROM NICOTINE IN'AGED LEAVESOF NICOTIA,VA TABACUM.. ..eata, Department of Chem- istry;. UnlYereity.afMlnnieeota „ Ninne- ep011s, MN 55455 9':20' 125 DETERMINATION' OP NICOTINE, NORNICOTINg, AND OTHER.SECONDAILYAMINE'ALKALOIDS'. IN TOBACCO. B. N. :ArdOn and G. H. Cree.ne, R. JI ReynoldaTObacco C'ompany, Research and:Development, Winsto.n-Salem,. NO 27102 9~:40i /26. CONFORMATIONAL.ANALYSIS'. OF NICOTINESALTS:.T'. A. Perfetti„ R. I. Reynolds Tolbacco Company, Researchland Develup- menr, Wlhscom-Salem, NC 27102 10:W $27 pD DEPENDENCEOP THE. CARBON:-1D NUCLFAR MACNET'IC RESONANCESPECTRUM OP NICOTINE.. R. F'. Stdven~. Lor111srd Corporation,. a~Divialbn of: LoeRaTTeatres,..Reseerch De~parcmenc,. Greensboro, NC 2742D 10:20' INTERMISSION Session B Hyatc House Regency3'.and 4 - C. J. Rosene;. Preslding 8':40i a32' ARAPID~METHOD,FOR THE DETERMINATION OF'. .N-NITROSONOR.VICDTINEIN C.IGARETTESMOKE'BY CLASS CAP'ILLARYGAS CHROMATOCRAPPiY.. N.: J:. ChambarZain, J.,L. Baker, and R. F. Arrendaie, Tobac-S'.afecy ResearcH h'nit,.USDA, SEAJAR„ Athens, GA 30613 9':00i /33'. CONCllRRENT GASCHROMATOGRAPHIC DETERMINATlON. OFVOLATILE', NONVOLATILE, AND TOBACCOSPECIFIC N-NITROSAHINES.. K. D.. Srvnne.m~nand D. Ho.ffmam, Amerlban Health, Foundatlon, V'alhalla, NY 10595 9':20' 134. APPLICATION'. OF'A SELECTIVE POLYMER: FOR RETIOVALi OF `IITRATE FROM TOBACCO!. P. R. Rhodes. Depe.rtmemc oE'Agronmmp., University.of Harylknd, College P.ark, HD 207A1, and J. A. SaunGers, TobaccoResearch' Lebeaatory., VSDA, SEA/AR, Be1[sville, liD20705'. 9:40' R35. APPLICATION: OFTENAR TRAPPING TD: T.HE ANALYSIS OF GAS PH'ASEORGANICSIN CIOARETTE5MOKE: C. E. ,q:ygins, W. H; Criest, and G. 0lerich, Oak RldgeNaclonal Laboratory, Analyti<al' ChamSstry' Divlsion., Oak' Ridge, TN 37830 10:00' /16ISOLITSON: AND IDEATIFICATION OF TWO,NES1. SECD-CFJOIRANOIDS' FROM:C.IGARETTE SMOKE. . Kiata, R. N. Fergyson,.R:..D. Kioser, andV. N'. Elnoli„ Phallp Norrls Resaerch Center, Richmo.ud, VA.23761, and T. P..Pitner, Baebrlhrvger Ingelheim, Ltd.,. Ridgeffeld! CT 06877 10:50' /28NETi.CEMBRANOIDSPRON BU4tLEYTOBACCO. 10:20INTERMISSION V. N'eemzn and U. Brummer, BAT' Clgarecten-Pabeiken GabH, Forecttung~ 10:50' /37SIMULTANEOUSRAPID GAS CHROMATOGRAPHIC. 1`.1D~ 29' und'Entvicklung, BahrenfelderChauseee 139, D-2000Hamburg50., Germany, FOUR: NEW CDMBRATRIENETRIOLS' FRON DETERMINASION OF WATER AND.NICOTINE IN'SMOKE.8'. N..('.ordan, R. ]..ReynolH.s Tobacco Company, Research and: Development, Wlnsron-Salem, NC 27102 TOBAC= :. WmhZbetg.,I. Walllh,. R. Arndc, K. Ndrdfore,.T. Nishlda, 11:10' /3B'A'RAPIDGAS CHROMATOGRAPHIC DETERMINATION andC.. R. Enre17„ Svediah' Tobacco Co~mp;eny „ Reaearch Deparcment,P. 0. Bo. 17007, 5-104 62Stock'holm, Svedln, OF MENTHOL.IN TOBACCO: B. M: Gordon aad M. F. 9orgerdi J, R:.J. Reynolde Ttrbacco Company„ Researchand. DevelGpmcnt,., Wi ' 11:30' /30CH'EY.ICAL COF@OSITIONOP DARK AIR-CURED ascon-5 alem, NC 27102 AND FIRE-CURED'TOBACCO. D. L. Davie, 11:30' f39'LOW MOLECULAR WEIGHT FATTYACID SuGAR ESTERS' :50' 1 B. H. Song, Tobacco andH'ealth ResearcT Irnstdtute and Department uf Agra omy, UnSveraltyofKen[ucky, LWxington, KY' 40506'; and V. Y. Weeks, CropScience Depprtment, North; CarolGna Srace University, Raleighi, NC 27650 CARBOHYDRATE CONTENT IN BURLEY TOBACCO :50 INTURKISH TOBACCO: SEPARATION BTREVEASE- PHASE HIGH PERFORMANCE.LIQUID CHROMATOGRAPHY AND SPECTRAL CHARACTERI2ATION. ':, M. R':-a, R.. J. Reynolda Tob'a¢co Company, Reses.rchiand. DevelopmenL, Wlheton-Salem, NC 17102 /40'DEVELOPMENT OF METHODOLOGY FOR', THE DETERMINA- TISSUE. E.. Leggett,. l. S'.ims,.and'~ J. Hemllton, USDA, SEA/AR, Department of Agronomy. University of Kentucky; Le.ingcon, KY 40546 TIONOPBROMIDE RESIDUES INTO:BACCO AND TOBACCO'SMOKE. PACTORS' AFFECTING'.THERESIDUE LEVELS RE'SULTRNG FRLRI:METHYL.B~Rq4IDE.PUMI- GATION. 4., Carn.mLght; A. Oorn<Ili, and V. Olertdcr,.Consolldated'Clgar Comppny; 131 OakStree.t,. Glastonbury, CTi 060J3 IV
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TWRSOATAFTERNOON Sesalon. A Hyecc Houae Regeneyl and 2 - Session BHyatt Houae Regencry 3'and. 4 - E. Glatk, L. D. Scory, Pteeiding Prealding 1':40Y41 DETERMINATIDN OPMOLYBDENUM IN 1:4D' 147 THIERMAL.DECOMPOSITION OF PECTIN. R:.R. BURLEYTOBACCO USING A'RAPID~ AUTOMATED METHOD. P. EiJasi, J, L. Sime, end J. C'rutchfleld; Depaztaent Squire and!R:.£. W¢jsCC.k,. Philip. Morria Research Cen¢er, P. 0. Bos 26583, Richmond; VA 23261 ofAgronomy, U'niverslty,of Kentucky,. 2:00/48THE.EFFECT OF SOME NITROGENOUSBLEVD 2:00 /42 Lexingcoo, Ky 40546 VARIATION IN'.MINERAL CONTENT OF COMHONENTSON NO1NOx AND'.HCN LEVELS IN MAINSTREA.YANDSIDESTAF.cMSMORE: V: !YOr~mn,, A.,N. Ihrig, T. M: Larsonand!B:..L. Mosa,. FLUE-CVRED TOBACCOS' DURING CURING. A. L.Scheparta, Tobacco Safety Lorillard Reeearch Center, Greensbero, NC 27420i :2oi tl43 Researth Unit„ and!R: L. Wllson,Biometrlcel Services,..USDA, SEA/AR, Arhena,.GA 70613, end J. F. Chaplln., Tobacco Research Laboracory, USDA, SEA/AR. Oaford, NC'.27565 THE CUTICULAR CHEMIBTRY OF GREEN. 2:20 f49 FORMATION AND TRANSFER.OF 4-(METHYLNITROSAMINO)-1-(3+PYRIOYL')-BL'TANONE(NNR). DURING SWDKING:. J. O..Ad, A. Castongiuy, S.,J. L'ee, N',. Vihchkoskf, and 0. Hoffmannj Amerlcan Heslsh Poundatian, Valhalla „ NY' 10895 TOBACCO.R. P. Severeon, R. F. 2:40 050 . PRECURSORSTUDIES' ON CATECHOL IN CIGARETTE 40 /44' Arrendete„ 0. 1. Chortyk, and LL g.Sm1rh;.TObaccoSafecyRea.archUdlt, USDA, SEA/AR; A[hena, 0A 30613~, J. P'. GhapLln end G. R. GVynn., Tobacco Research Lrboracory', USOA', SEA(Ag, Oxford, NC27565, and A..W. Johneon, Pee Dee Experiaent. Stacion., Flerence,. 50 29503 THE QUANTITA'IIONOPTHE'MAJDR SMOKE. S. Ca1.ne~IZG„ S. S. Hech[., andD'. Hoffmann, Amerlcan Health~FOUndatiom., Valhalla, NY10595 CIPfICULAR LEAF. COM@ONENTSOF GREEN TOBACCO. R. F. Seversoni,R. P. Arrandmla, 0. T.Chortyk,.K. U. HcDuffle„ and Pli:.F Naaon, TobertcoSafety'Reseerc,U'nic,. USDA, SEA/AR, Achena,.GA 30613, A. W:,Johnamn~.Pee Dee.ExperLment Stac.ion„ Floremce, SC.29503, N.. D. Jackeon, G. R:.GVynn,. and J. F. Chapll¢, Tobacco Resesrch Laboretory', USDA, SEA/AR, Ozford, NC 27565 3:00: INTERMISSION 3:30i45THE FRACTIONATION AND ISOLATION OF THECUTICULAR.COMPONENTS' OFOREEN'. TDBACCO. R. F'. Severson;.0. T. C7ertyk, and K. L. MeDUffie,.TObaccoSafecy'ReseerchUn.it., USDA, SEA/AR, Achens, GA30613, A. Y. Joh.oa, Pee Dee Experiment $tec3oa,.Floreoc., SC29503, M..D. Jacksom, C.. R. Gvym,andJ, F. Chaplln, TobaccoRe.earch',Laboracory., USDA, SEA/AR, Oxford, NC 27565 3'50 #.46COHPARISON'0F THESURFACE. CHARACTEI[I6TICS' AND CHFMI6TRY OPNORMALi ANID GREE'N. PF.bCH APHID RESISTANT TOBACCOS: R: F.. Seueraon, and K'.. L.,ycDuff3e, Tobecco SefetY. Reeearch'Unit, USDA,.SEA/AR, Athena„ GA 30613„ and'A.k'. Juhnson, Pee DeeEapprdm.oc S.tacioo, Fiorente, SC 29503' 4':30. BUSINESS MEETING: HYATT HOUSE REGENCY 3 end 4' 6':30: SOCIAL HOUR:. GARDEN TERRACE 7.:30' BANpUET:. HYATT HOUSE REGENCY 1, 2,..3, and 4 V 3'00 INTBRMISSION 3t30/51 OICARETTE PACK MOISTURE VARIAT'ION IN. STANDARD'SHfPPING CASES. X..A. FLh~ert,, grovn.and W1111amaon Tobetco Corporacion, Louiav111e, KY 40232' 3:50 /52A MODIFIEDQUALITYINDER OFFRESH. CIGARETTE SMOKE. J:-C. PQTK,,TL-HI. Lee,.and K;.-K.,Yoo, Kocea GSnseng and Tobacco Reaearch.. Insticute,.5eou1,.. Korea 110
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FRIDAY MORNING, October 9,1981 Joint Session Hyatt House Regency, 1,. 2', 3, and 4. - D. L.. Roberts, Presiding 8:45 Philip Morris Avardpe,.Research Highligh'ts9:15, #53 A METHOD FORCONSTRUCTINGHEIGHT DISTRIBU1TiIONS VS SHRED DIMENSIONS. M. E. Counts and B'. C. LaRoy, PhilipMorris Research Center, Richmond, VA 23261. 9:35 i54' A METHOD FOR MEASURING THE AREA OF TIPPING.PERFORATIONS. P. F..Perfettl,.R. J. Reynollis Tobacco. Company, Research'and Development, Winston-Salem, NC 271029:55/55 CALIBRATION AND CORRELATION OF TAOTYPES'OF CORESTA PERMEABILITY METERS.. A. L. Bens.on, T. Kenefick, R. McCarthy, and:P. Sutherland, C. H. Dexter Division, 2 Elm Street, Windsor Locks, CT.0609610:15/56 COMPllTER STUDIES ON THE RELATIONSHIP OF FLAVOR TO MOLECULAR SHAPE..,R. S. Mamr.or, Lorillard. Corporation,.aDivi'sion of Loews Theatres,.Resea:rch Department,.Greensb'oro, NC 274201 10t35. i57THECONCEPTS OF FLAVOR INTH.E CIGARETTE INDUSTRY..R:.AbdalZah, A. C. Mon.k & Company, Inc., Farmville, NC 278288 lOt55CONFERENCE ADJOURNMENT VI
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WEDNESDAY MORNING 8:15 OPENING REMARKS AND WELCOME 8:15 SYMPOSIUM: TOBACCO LEAF CHEMISTRY: ITS ORIGIN, UNDERSTANDING AND CURRENT TRENDS Lance Reynolds, Symposium Chairman -1- 8:30 TOBACCO LEAF: "ALOOK ATiHISTORY.°I. E. Massie„ College of Agriculture,. University of Kentucky, Lexington, KY 40506 Tobaecoo apurely. American plant,. dates back eent~uri'es. The "discovery" of the leafi's credited toColumbus when he "found"the NewWorldand saw the natives - Indians - using.tobaceo, The plant was first thought to havemedicSnal properties and for years was proclaimed a"miracleherb"". But the medicall aspects were eventually discounted and use of the plant has long.been limited to,creature pleasure use - for smoking, chewing and snuff. The historyy of the plant r~evealsits importance in war and peace and its economic impact in the development of the United States - aswelli as the economic importance in the present. England was the.first major export market for American tobacco, but during the Revolvtion.worl&tradein tobacco began its initial expansion, Controversy over the use of tobacco began almost from the outset, still continues, and is yet to be resolved. The tobaceo,industry has remained basicalilyunchanged from the early cultivation of the crop, but refinementstiave been,intro-duced and quality advanced. There have always been problems, and:theyy still exist in the industry. Problems have become more complex, involving suchh things as credibility, health, and production. Organization is needed in the industryy to face these.problems. Unless change is made, the industry will continue to face mounting problemss in the future. 9:00 I CHEMICAL AND PHYSICAL CRITERIA FOR TOBACCO~LEAF OF MODERN DAY CIGARETTES. A. W. Spears and. S. T. Jones, LorilQiard:, 25:25E. Market Street.,. Greensboro, NC 27401 New cigarette brand introductions and brand~extensions have resulted insignificant changes in the average product sold over the last five years. Extrapolation of these recent product trends indicates that an inereasinglyslgnificant change will be taking place on a sales-weighted basis. This evolution toward reduced tar products has impor- tantly influenced, and will continue too affect, the average chemical and physical prop- ties of the tobacco blend'sthat are being used. Advances in analytical instrumentation~ have allowed scientists to explore and better understand the s2gnificanceof thesee chemical and,phy:sical properties. This paper reviews trends in the usage.of tobacco~ types including reconstituted sheet. Additionally, the processes for expanding tobacco have produced a trend toward lower blend densities.. These trends, both historical and future, are diseussed.In~support of the text, information on the usage of tobacco types and'their chemical and physical properties are presented. Where.appropriate to the chemical or physical eriteri'a, eonsideratdon is given to individual tobacco constituents including those components that.are generally recognized as important to,the organoleptic qualiity. Trace components, and more particularly.residues, are reviewed with respect to their importance to the quality of modern dayleaf'_. 1
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WEDNESDAY MORNING - SYMPOSIUM -3- 9:30 i MAJOR CHEMICAL CHANGES DURING SENESCENCE AND CURING OF TOBACCO LEAF.. R. C.. Long.and J. A. Weylirew.Crop Science Department, North Carolina State University, Raleigh, NC27650 Chemical changes which occurdurimg senescence and curing are common~tlo all tobacco types, the magnitude of the changes being dependent upon the curing regime and the coneentration.of initial'substratescondiitioned by genetic compositionn andlgrowingenvironment. Theeomp,lexityofthe biological syste-makesit'.difficullt to ascertain the onset of senescence in the tobacco plant. Unlike the climacteric infruit,.theshift in metabolic actYvityin tobacco leaves during,growths maturat~ion,.and senes- cence is more grad'ual. For example, the.eessation of expansive growth occurs coin- cident with declining nitrogen uptake and reduction~and with the onset of starch accumulation after about seven to nine weeks' growth. Thesubseqpent senescence of physiologically.mature leaves is characterized visuallyby'degrad'ation of chlorophyll and carotenoid pigments, the hydrolytic products perhaps contributing to the flavor of the tobacco. Those pigment losses are accompanied by degradation of starch and chloroplastlic proteins (principally ribulose bisphosphatecarboxyiase)i. Chemical transformations during curing are b'asicallyextensions of those.occurringduringsenescence but proceed at accelerated rates during,the yellowing phase. Certainamino.aeids accumulate and othersdim~ihish. Simple sugars accumulate but may bec.onsumed by, respiration depending,upon the time of termination of biologicaL activity. The inf'lueneeofe leaf yellowing agents and accelerated curing sched'ules on these transformationswil'1 bediscussed.. 10:00 INTERMISSION -4- 10:30 METABOLISM AND~REGULATIONOFALKALOIDSIN TOBACCO. L. P. Bushs I Department of Agronomy, University.of Kentucky, Lexington, KY 40546 The biosynthetic pathways for tobaccoo alkaloids - nicotine~ nornicotine,.anabasihe, anatabine and myosmine - have become accepted in principle in precursor-product r.elationships. Enzymatic and non-enzymatic reactions in the biosynthetic.sequencearereviewed with respect to rates and tissue origin. Rates of growth and develop- ment of the plant' alter nicotine.accumulat.ionand enzymeactivities,e but the regulat~ion of alkaloid synthesis and/oraccumulation is not understood. Analysisof.the domSnant~ and recessive alleles at the tw-princip,al loci controllingalkalo.idg accumulation indicatesthat thedomihant alleles at both loci',areassociated witti~ higher Levelsoftotal nitrogen and total volatile nitrogenousbases„ but lower levels of protein nitrogen than the recessive alleles. Nicotine accumulation in, callus or suspensi-cuLturesof Nicatiana species appears tob'e variable and! influencedib'y genetic predisposition, nature and rate of growth, liighting„ and growth regulator combinations. Cultures with~either high or negligible nicotine concentrations are obtained by, proper selection of cell strain and growth condition. It has been shown that exogenous nicotine acts as an induction factor in morpho- genesis im.tobaceo~tissue culture. It has been proposed that endogenous nicotinee may function also in morphological changes. 2
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WEDNESDAY MORNING - SYMPOSIUM -5~ 11:00 THE ROLE.OF TOBACCO LEAF PRECURSORS. IN CIGARETTEFLAVOR'. R..A. Heckman4 M:, F. Dube, D. Lynm„ and J. M. Rivers,.R'. J.. Reynolds Tobacco Company, Research and Development Department,. Hinston-Salem,. NC.27102 Various aspects of the relationship of tobaceo~leaf and smoke composition to smokingqualityg have been dlLscussediat recent symposia. In the present reviewli~ttleattention is given tochemieal changes.that take plaee.duringmaturation and'post-harvest handling.oftobacc.o.. Of pr~imaryconcern are those smoke components considered to ariseprdmarilyby, pyrolysis of non-volatile leaf precursors and which are likely, to.have flavor impact. Acfiromatographic comparison.of'~ leaf and smoke volatiles is made to provide visual confirmation~.ofthe increased complexityy of smoke.composition~over that of tobacco. Various leaf'~ flavor precursors are discussed in.terms of the numbers of volati'Sesmoke eomponentseach precursor mightt contribute to smoke. Nitrogen..and sulfurr smoke components arer~eviewed, and~ evidence for the prob'abDe origin and flavor potenti'aI of; specific pyrazines, pyrroles, pyridines, and heterocycliesulfur compoundk.is presented. The role of sugar esters, glycosides, Amadori compounds, and lignin in the development of cigarette smoke flavor is also discussed. -6- 11:30 TOBACCO LEAF PROTEIN: MOLECULAR.BTOLOGY'A.ND GENETIC MANIPULATION. S. D. ICung and T: C. Tsoy Department of Biological Science.s,.Universityof. Maryland Baltimore County.,,Catonsville, MD 21228 and Tobacco. Laboratory,. Beltsville Agricultural Research Centery,SEA/.AR„ Beltsville, MD 20705 Fraction 1(F1):protein is the most abundant solubleleaf protein ingr~een plants. In tobaceos F1protein accounts for moree than25X'of total leaf! protein. This protein is an important.photosynthetic enzyme, ribulose 1,5-b'isphosphat~e carboxy.lase- oxyg2nase (RuBPCase). IR is unique in.many aspects. It'catalyzes.two,r~eactions: photosynthesis and'photorespiration, consists of two.t~y,pes of subunits: 1'arge and small, and requires twoseparateo genomes for itss synthesis: chloroplast'and:nucllear. Tobaceo.fll protein.can be crystallized by.a.simple procedure with high yield. The pure enzyme consists of eight large andd eight small subunits. Each large subunit (LS): contains separate sites for activation andicatalysis.whereas,the precise.func.tions of thesmallisubunit. (SS) aree still unknown6 Tihe LS,is coded for by chlbroplast genomes,and synthesized,on,70SS ribosomes vRiletheSS is coded for by nuciear genomes and synthesizedon,80Sd ribosomes. SS is synthesized as a large precursor whicii~is subseqµentlytransported'and processed before it is united with the LS to form,a funetionalienzyme.. Generally,.the, composition of large and small subunitss can be genetieaily, manipulated in such a way that enzymes consi~sting,ofidencical LS and di~fferent SS,.orvice versa, can beconstructed... With recombinant DNA technology, genescoding,f~orthe LS and SSc.an be isolated', clonediand expressed in vivo.This.of~fersan excellent opportunity.for genetic engineering of tobacco FL protein for the: purpose of obtainingbetter quality.and higher yield. 12:00 SYMPOSIUM SUMMARY AND FUTURE PROJECTIONS - Lance Reynolds 3
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WEDNESDAY AFTERNOON - SESSION A Technical Session A: J. M. Elliot, Presiding -7- 2:00 THE CULTURAL MANAGEMENT OF FLUE-CURED TOBACCO STEAM-VOLATIliE CONSTITUENTS. W:. W. Weeks and J. A. Weybrew,.North Carolina StateUniversity„ Rale.igh,. NC 27650 Proper management of fliue-cur~ed tobacco is the key to success in producing a qpalitytobacco product. Nitrogen fertilization „ soil type, cultivation moisture, and curing greatlryinfluence the physical and ctiemical propertiesofs the leaf. The effects of such managements on the neutral volatile ails from.cv. NC.2326 grown~ dur.ingtrie. 1980 season at Central CtopsResearch Station„ Clayton,.NCare presented. Fifty-twovol'atile constituents fzomseventy-two samples.were compared chromato- graphieallyy after steam distillation.Signif'~icant differences (P <.0.05)assoeiated! with particular management variables were measuredL Simple.correlation coefficients, r,, between pairs.ofconstituents were computed as clues to metabolic relationships, biosynthetic intermediates, and/or degradation products. Manyy of these correlations were statistically, significant. The magnitude of significance among.constituents and treatment effects will be discussed in.detail for manyof'~ these:compounds. -8 ~_ 2:20 BIOMASSAND CHEMICAL COMPOSITION OF TOBACCO PLANTS UNDER'HIGH DENSITYGROWT'H~ S.J. Sheen,.Depar:tment of Plant Pathology.,, University of Kentueky,. Lexington, KY 40546 Tentobaceo~culitivars representingvardous types were grown at a density of 75 to 86 plants per square meter and were harvested at knee-height. Biomass yield ranged from 44 to 70 metric tons per hectare wilth Ky 16 Mammoth the highest. The Mammoth variety, however, had the lowest leafi/stem ratio (1.7.7).,,whereasCoker109was the highest (4.04). One metric ton of biomass would produce.about 6kgofsolub'le proteins, which is independent of thee leaf/stemratio.Analyses of the deproteinized fibe.rs.revealied tFiatt the levells of neutral and acid detergent fibers, cellulose and hemicellulosewere comparable among the ten cultivars. A large variation of li.gnih eont.ent.was observed among the dark tobaccos. Burleys, in general, had lower concentrations of starch and insoluble proteins ihthe fibrous residue than did the dark tobaccos. The present resulRs suggest that the.selection~of tobacco genotypes is of importance for high density cultivation in~the production-f soluble proteinsf'~or human consumptlonand f'ibrous resid'ues as animalifeed. 4
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WEDNESDAY AFTERNOON - SESSION A -9- 2:40 OONTLNUOUSFLO4l SYSTE.M FOR.THE EXTRACTION~OF PROTEIN' FROM TOBACCO. LEAVES. DiE. B1ume„ W. G. Woodlief and J.. J. Lam, USDA, SEAfAR', Tobacco~Research Laboratory.,.Oxford,.NC27565 A procedure has been developed for the crystallization of ribulose 1,5-bisphosphatecar~boxylase (Fraction I Protein) and subsequent recoveryof'. Fraction II Proteins from homogenized tobaceo~leaves. The continuous flow.system.isadaptableto large-scale protein removall and does not require high salt concentrations,. Sephadex columns or fleat' treatments toobtain,protein, Freshly harvested tobacco leavess are homogenized and the slhrry is pressed through a nylon filterto retain fibers an,ice.llular debris. The green filtrate ls processed througH..a 0.2-micron cross-flow filter which removes chloroplastic particles. The protein permeate is pumped t'hrough- series of' hollow fiber cartridges to.concentrate the solution and selectively collect proteins;of varying molecular weights. Protein fractions are then.processed in a diafiltration mode witFi~a buffered solution to remove contaminants and assayed for RUBPP car6oxyliaseactivityand total protein.content prior to drying.. -10- 3:00 UPTAfCE.AND INCORPORATION oFjH'rL'EUCINE B'Y GERMINATING TOBACCO.SEEDS. J. Ar~cils, S. C. MoHapa[raand W. H. Johnson. Departments.of Botanyy and'Bio. and~Agr..Bng., NCSU6 RaLeighs NC 27650~ During tobacco (,Yt tabaewrr L., cv Speight G-28)) seed germinationy no morphological change takes place during thefirst 48 hours of im;bibition. This developmental lag isnot relatedd to moisture limitations because moisture saturation isat'tained by, six to nine hoursfolLowing.soaking. Furthermore, the fact that maximum sensitivity to light develops during the fitst 48 hours of germinationindi~catesthat one or more subcePlular eventsmust be taking place during thi's period. Studies on dry weight, osmotic potential,, and'~respiTation did not reveal any detectable change in these physiological parameters.. Eleesronmicroscopic studies also did not reveal any ultrastructural change. Th'erefore, chan4esat molecuilarr level were investigated through pulse labeLling.oftritiated (,H). leucine at' 25°0 under continuous ililumination. This label was incorporated to theprotein:fr,action~ TCA-insoluble fractionYmaximum during the first 24 hours of imbibition. Thus, it seems,that ibspite of the 1'acK oCmorphmgenetieand physiological changes during,the first 48 hours of germination~ changes.involving proteins,and'perhapsothez biochemical components, occur prior to,the subsequent growth process. The regulatory signifflicanceof ofthese evenwill be discussed. 3:20' INTEAiMISSI'ON 5
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WEDNESDAY AFTERNOON''- SESSION A -1Q- 3:50 COMPARISON!OF THE DEVELOPMENT OF MAJOR LEAF COMPONENTS IN NORMAL A.1`D.A. BCIDWORM RESISTANT TOBACCO. R.F, Severson, 0.. Ti. Chortyk, M, E.Snook, K. L. MoDUfifie, and L. B. Smith, Tobacco Safjety.Research Unit,. USDA, SEA/AR, Athens, GA 30613'and G. R: Gwynn, T6bacco.ResearchLaboratory,. USDA, SEA/AR, Oxford, NC 27565. Numerous reports have shown~that Tob'acco. Introduction TiL-1112 has a high degree . of resistance to tobacco budworm. Recent studies indicate that the observed: resistance may be due to.oviposition preference. However, the young.budvorm also fails to feed on~t~he young,leaf of TIi-1112. It wasofihtere.st to.determineif'o wholie leafi chemistry.maybe responsible for this lack of feeding stimulus. TI-1i112' and two non-resistant varieties, NC 2326-nd Speight G-28, were sampled at various stageso.f~ development from bud'leaf through curing. Samples were.fr~eeze-dried where required in order to stop further enzymatic changes. Aliquots of ground samples were mixed vith.BSTFA/DMF ih smalli reacGi-vialsand analyzed „ afterr our one-step ultrasonic extraction-derivatization.pnocedure, by.gas.chromatography on a.short packed Dexsdll 300.GC column. In this initial report we will.present the changes in malic, quinic, and citric acids,, changes in the.f'ree sugars (fructose, glucose, and sucrose), solanesol, and the polyphenols in the growth of a mid-stalk position leaf. Individual leaf'~ polyphenols were aiso.determined by HPLC. The possiblie relationship of leaf chemical changes.t'.os insect resistance will be discussed. -12- 4:10 DIALLEL GENETIC ANALYSIS OF LEAF AND SMOKE CHARACTERISTICS IN FLUE-CURED TOBACCO.R: S.. Pandeya andlB. F. Zilkey,.P. Oi Box 186,. Delhi, Ontario, Canada N4B2W9,,V. A.DiIrksy Harrow, Ontario, Canadg NOR 1G0, and C. Poushinsky,,.. Centrali Experimental Farm,.Ottawa, Ontar.io„ Canada. K1AOC6 A five parent diallel set (excludingreciprocals), was evaluated for variation in aa number of smoke and'leaf chemicalicharacteristics in flue-cured tobacco. The range ofvariation.in all of~ these (except for pressure drop and weight of tobacco/cigarette)~ wasconsid'erabLe.and. ANOVA suggested the presence of significantt genotypic variance.. Statistical treatmentt ofthef data by Griffing's Model II, Method II analysis indicated significant variance due to general combining ability with almost.no.genotypexyear int'eractions.. General eombiningabili~ty estimates exceeded specific combihing,abiRityvalues in most cases indicatingthe major importance of additive genetic variance (ob '. in~controlling TPM, nicotine, aldehydes,, wet tar on a per cigarette basis as welil'..aspercent ash,.total nitrogen „ and cellulose in the.leaves- Wet tar showed a large measure of non-additivity; this may be associated with pereenG nicotine transfer and the two extreme alkaLoidleveliparenGs, Lonibow andiD2. Thevar.iance components estimates of general (o2gca) and specific combining abilities.(cZsca)vereoften identical'.for the.tvo years. Our.results indicate that selection for the above cigarette and lieaf vardables,should be possible. The ozg,cyand o2sca results have been compared with the Hayman's Wr-Vt graphic results and genotyp.ic associatfonsbetween smoke and~other variables.havebeen characterized... 6
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WEDNESDAY AFTERNOON - SESSION A -1d- 4:30 PARTICULATE MATTER VARIABILITY IN BURLEY TOBACCO. D. L. Davis, G.. B. Collins,andP. Di Legg, Tobaceo,and Health Research Instit:uteand Departmentt of Agronomy, University of Kentucky, Lexingtons KY~ 40506 The ob'jectivesof this research were to determine thee effect of genotype, Leaff positions andeultural practices on total particulate matter and!to,examine~the relationship between particulate levels as determined by standard smoking procedures~ and total particulate matter index (PMI), as measured by pyrolysis ofground tobacco.. The average PMI values.r~anged~from 182'mg/g of tobacco,tlo 218mg/g., There was not a wide range of valuesfor; current commerci~al burley.varieties. Low alkaloid genotypes had reduced particulatematter del'ivery: Therewas a significant difference in PMI and. TPM among stalk positions with.lower staLktobac.cos yielding lower values as expected. Increased.field'pop,ulations did not si~gnificantly reduce PMI in two burSey eulbivars with differentt leaf angles. It would appear that.PMI could be used as a methodd to eompare.particulate matter yield if proper sampliingand laboratory conditions are followedL Therevas an inreraction,between~stalk position~and method.. Nicotine levels were~measured on the filter pads obtainedd from the two procedures with.satisfactory agreement in the data. -1!4- 4:5D MOISTURE SORPTIONPHENO'MENAINICURED TOBACCO. T. Samejima and'Y. Nishikata,: CentraL ResearchInst~itute, The Japan,Tobacco and Salt PublicCorporation„ 6-2'Umegaoka, Midori-ku, Yokohama, 227 Japan The dynamics of moisture movemenc in acur~ed tobacco tissue is of practical interest in connecti'on with drying.or wetting processes. The pathway and the.rate of moisture movement were investigated to analyze the moisture diffusion~mechanism~.in tobacco... Experimentswere conducted.with laminarr andshreddedtdssuesfromflue-cured tobacco (8Y-4);.. The.moieture sorption rate (mg/cm2/sec): through the eut' face was much faster than~the rate.through:thee leaf surface. Sihceshreddedtissue has a large.cut face area most of the moisture was ab'sorbed'through the cut face. On thee other hand,., in laminarr tobacco,, most.ofthe moisture was absorbed through the leafl surface. The moisture sorption~rate through the leaf surface became extremely.fast when thee epicuticuiar- layer was removedd wit}ichPoroform. This-r,esult suggeststhaxthe moisture sorption rate in th'elami'nartobacco:is strongly controlled.bythe rateof vapordiffus.ion through theepicuticular Layer. The rate of moisture sorption can be.descrl6ed by thefolilowing.empirical equation:: dW/dt'= K(We>W)n;~wriere uis the average moisture content at time t; K..and'nare empiricali constants; and We is the equilibrium moisture. content at a.given condition. The mechanism of moisture movement will be discussed;on..the.basis of the.mass transfer coeffdcientss through the leaf surface and~the cut face of tobacco in relation to environmental conditions.
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WEDNESDAY AFTERNOON - SESSION B Technicali Session B: C. L. Browne,,Presiding -1s- 2:00 COMPONENT ANALYSIS OF CIGARETTE VARIABILIIY. D. E. Mathis„ Tennessee Eastman Company.,,Research Laboratories, Eastman Chemicals Division, Eastman Kodak Company, Kingsport, TN'37.662 Theoretical equations arepre.sented whichrelate.the.variabflityh of vented cigarette dilution and drypar.t.iculate matter (DPM)~deliveriesto the pressure drop variatiilityy of the filter„ tobacco column, and;vents. These equations arederived.by applying, statist.icai re.lationshipsfor the calculation off variancee to well documented equations describing dilution and DPM delivery~ Simplifying assumptions requirediforthederivat.ion are stated and shown to have an insignfYicant effect on the aecuracy,ofthemodelwhen.ventilation levels do not exceed50L,, The performance of the model is demonstrated by, comparing predictediand measured values of dilution andIDPMM delivery variabilityfor experimental cigarettes. Application of the model for~ determining major sourcesof: cigarette variability and themoste effective means of: improving uni~formitg, are pointed:oun.Isovariation.contourgraphs are introducedd asa useful meansof'~ displaying ttie.predicted dependence of venti~lation.and DPM delivery variability on the~variability, of the cigarette components. These: plots permit filter and ventsystem~pressure drop variability limits to be established! whieh~achieve.aspeeified variance.in~DPM delivery, and ventilation. A long term obj,eetive of this study, whicri~has not yet been~realized, is to incorporate cigarette uniformi~ty and resistance to component variabilityy as criteria in,the de.signofventilated filterr cigarettes. -1'6- 2:20 REDUCED EFFICIENCY VENTILATED FILTERS PROVIDING LOWERED OARBON MONOXIDE PERUNIT' TARDELLVERED. P..J.. Bohlander, Celanese.Fibers Marketing Company, P. 0. Box 32414, Charlotte, NC 28232 It has been~wellL dbcumented'that filter tip ventilation~has considerable influence: on the amount of carbon monoxide andlotherr gas-phase components delivered from.a cigarette. Unfortunately,,, lowered carbon monoxide deliveries achieved with filter tip ventilation generally dictate reduced taste and impact as well, resul'tingfrom nearlyequivaient reductions in particulate delivery. The objective of this workk was to investigate filter designs.which use.ventilation to reduce cigaret'tee carbonn monoxidedelifvery while still maintaining a,high level of particulate delivery. This was achieved by, substantially lowering the particulate removal efficiency. (perr unit length and cikcumference) of ventilated filters which increasesthe.ir tar/carbon,monoxidedeliveryratio over that routinely obtained at equivalent levels of ventilation. Physical changes.in filament denier and cross section~ along,withvarioussmoke channeling techniques were evaluated as means of lowering the filter parGiculateremoval efficiency. Increases in thetar./carbon monoxidee delivery ratio.at'equivalent ventilation levels wereexhibited byy all the reducedd e.f.ficiency,filters.These results along with..subjective tasteevaluations of thereduced:effic:iency f:flterswil'L be discussed. S
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WEDNESDAY AFTERNOON - SESSION, B -17- 2:40 CIGARETTE PAPEREFFECT6 ONTAB/NICOTSNE AND CO/TAR.RATIOS. A. McMurtrie,. E'. F. Litzinger, and D. T. Wu, Brown and Williamson Tobacco Corporation4 1600 W. Hi1~1 Street,.Louisville,. KY 40201 Paperr permeability and citrate content effects on.tar/nicot'ine.and CO/tar ratios for a set of 27 cigarette samples have been examined. All samples were made to the samespecifications, differing only in cigarette paper.. Tar and nicotine values were obtained by conventional procedures. CO deliveries were determined simultaneously with tarr usingnondispersive infirared'.analysis. A wide range of paper permeabili,tiesand citrate levels wereexamined. Theresults.o.f'e the studyy indicate that tar/nicotine ratios are determSned'primarily, by.paper permeability; high permeability produces the lowest.tar/nicotine ratios. Paper citrate has adefihite but.much smaller, effect on tar/nicotine ratios at bothimedi'um and 1oww permeabilities,, but appears to have little or no effect with high paper perme- abilities. On the other hand, CO/tar ratios are determined primarily by paper citrate content with'ilow citrate produ.cingthe lowest CO/tar ratios. Paper permeability has a def~inite.but less pronounced effectt on CO/tar ratios. Th'eres.ul~ts of thestud.ye also show that thosecitrate-permeabilitycombinations whieh~ producethelowest tar/nicotine ratios overlap very, little with those combinations producing the lowest CO/tar ratios., -18- 3:00 THE.EFFECT OF CIGARETTE PAPER POROSITY,AND PREPERFORATED TIPPING PAPER HOLE SIZE ON CIGARETTE VEN'TILATION.AND SMOKE. DELIVERY.. R'. A.Hyd'e, CellaneseFibers Marketing.Company, P. 0. Box 32414, Charlotte,.NC28232 Tipping paper permeability and plugwrap porosity are two variables most commonlryc.onsidered in the ventilation of a cigarette.However,.twoadditional variables, cigarette paper porosity and preperforated t.ipping.paper hole size, have been found to have a significant eff~ect.on~total aigarettedilution.and smoke deliverry..Cigare.tte papers,,varying in porosity from 25 to 200 CORESTA unit's,., were evaluated with.f'iilterconstructSonsventilatedh from 20 to 80%. It'was found that a linear relationship exists betweentdp dilution andt~otal cigarette dilution for any given cigarette paper porosity with'.the slope of the line dApendent upon cigarette paper porosi~ty. The effect of varying cigarette paper porosity on.smoke chemistry was also evaluated. To determine the effect of preperforated tipping paper, hole siie on filter dilution and smoke deliveries, micro and'macrolaserd perforated tipping papers ranging.in permeabil'ityfrom 4001to 4000~CORESTAuni~ts were evaluated with filter constructions ventilated from l0ito 80%. By.varying tipping pape.rholesize at constant permeability, it was.found that the effectivearea of a filRersubmd[ted to ventilation can have a significant effect on the amount of filter ventilation achdeved. Since smaller perforations require more holesto achievee the same permeability leveL than large holes, the effective area of the filter exposed~.oo ventilation is larger, yielding.a higher percent dilution. The effects of tipping paper hole size on smoke chemistry was a1soo evaluatedL 3:20 INTERMISS'ION' 9 I
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WEDNESDAY AFTERNOON'- SESSION B -19- 3:50 THE EFFECTS.OF VENTILATIONAND~PRESSURE DROP ON FI!LTRATIONEFFICIENCY, R. W. Dvyer, S. C. Ab'el,and;.M. L. Fleming, Philip Morris U.S'.A_ . Research Center, P. 0. Box 26583,. Richmond, VA.23261 A model of aerosol filitration based on the properties of the smoke aerosol and the filter has been developed., The properties of smoke accountedfor~ are its particlee concentration and ve.locity; the filter characteristics considered~inc.Lude length and pressure drop. This model has been derived from filtration efficiency data for bot}iCA filters and cut-tobaecorods. We havefound~that the filtration efficiencies of conventional CA filters at the same pressure dropars equivalent at a given f:low.rate„ independent of the tow item. The theoretical basisof~ this model'involves semiempirical treatments of diffusion, impaction, and interception~ mechanisms of particle capture. The relative contributions of these processes were evaluated within the f~ramework.of the theory. Tihe res.ulits predict.thatt diffusion is the major aerosol removal mechanism in filters, while impaction predominates in tobacco beds at tFiee standard flow rate. A.more easily.applied method is provided for those concerned with estimating the efficiencies of ventilated filters... This techniquee allows onetoc.omputethe TPM efficiency of a filter based~on.itspressure drop and initial ventilation level. Results predicted byy this technique agree well with the experimental data. -20- 4:10 APOLYDISPERSE'AEROSOL MODEL FOR CIGARETTE SMOKE FILTRATION. D. D:_ McRae, Philip~Morris UL S. A., Research~Center, P: 0. Box 26583, Richmond, VA 23261! Awelil-developed'modelifor filtration by.fib'rous-bedlfiltershas been-odified to predict filtration efficienciesfors tobaccoo smoke in cellulose acetate cigarette filters. The modif'dcations includet~he ad'dltion of a flow.rat'~edependent term in the solution for the stream~function.that corrects for the relativelyhigh gasvelbcities found in cigarette filters.Aiso; a technique for caLculating the.over- alli efficiency for a polydisperse aerosol, such as cigarette.smoke partlcles, was developed. The tobacco smoke particles were assumed to have a geometric mean, radius.ofi 0.10 um-i2ha.logarittimich standard deviation of 0L35.. The overall efficiency, was.determined byy calculating the penetration for each particle size, multiplying it by themagpitude of the distribution f~unctionf'~or that size, and then reconstructing the size distributiono8 the exit't smoke. Filter e.f#iciencies calculated by this new.model' mode1excellent agreement with.theexperlmental data for alvarletyof ciigarettee filters over a wide range of flow rates. The model and its derivation are discussed. The predicted dependence of the eff'~iciency on various fi~lter parameters such as f'~low rate,, filter length6 packing density, and fiber size arep.resented along with the relevant experimental datafora comparison. The theory, when combined with a me2hodifor predicting pressuredtop, can be used to predict theef.ficienc.y for any hypothetical CAfllter. Also,.the theory can~estimate the relativee contribut.ionsof the various filtration mechanisms and shifts in.che sizedistribut~ion of the smoke due to thee filtration process. 1QI Z I I
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WEDNESDAY AFTERNOON'- SESSION B -21- APORESIZE MEASUREMENT TECHNIQUEFOR THE'CHARACTERIZATION OF CIGARETTE 4•30 COMPONENTS. D. E. Mathisand C. H. Weatherly.,,Tennessee Eastman Company, Research Laboratories„ Eastman Chemicals.Division, Eastman Kodak Company, Kingsport, TN:37662' A pore size measurement technique was developediand applied to the characteristics off cigarette components, including filters, porous plugwrap, and.perforated tippingg paper. This teehniquewas adapted from ASTM F 316-70 (1976).,which was originally developed f~or measuring the pore size of inembranefilters. The method consists of determining flow as a function of pressure drop for the sample both in.the dry state and after being wetted with an~appropriatenon-solvent liquid. Sinee.thepressure requiredito displace liquid firom,..the poresofithe sample depends on.the pore diameter,, it is possible to determine from these measurements the percentage of the total f~lbw.asa function of poree siie~ The pore sizedistrib'ution determ~ihed by this technique is,weighted, therefore,, by flow ratherthanbyr numb'esr of pores and is moredirectlyrelatedltosuch relevant properties as pressure drop and permeability. The methodd alsoallbwsthe determination.of a maximum and median pore diameter. The use of the techniqUe for the characterization of flowthrough celluloseaeetate filter tips and tipping paper with mechanical, electrostatic, and~laser perforations is presentedd and discussed.. -22- HIGH SURFACE AREACIGARETTEFSLTERS. C. H. Keith, Ce.laneseFibers 4:50i Company, P. 0. Box32414,.Charlbttey NC 28232 In thefiltration,ofe tobacco smoke, it has.long been recognized both.experimentallyand theoreticallythaty increasing the surface area of the filter increases the pressure drop and removal efficiency. However, the increase.in pressure dtop can be avoided by altering the filter construction so that the high surface area is lbcated in areas adjacent to but nott direct~lyy in the main-moke stream. This paper illustrates this point by, comparing filter construction containing an experimental high surf~ace area fibrillated eellulbseac.etatepulp. In aa normal filter construction, with the material attached to the surface of ceilulose acetate filter tow, increases in.both pressure dtop and removal efficiency are encountered. Ihaeorrugate&sheet construction, comparable amounts of the fibrillated material and acetate staplee giNe an increased removal effieiency.without.thepenaltyofy increased pressure.dtop. The properties of this pulp and the fabrication of both,types of fiLtersare: discussed. The physical properties of these filters and;the deliveryy of particulate matter, nicotine, water, and nicotine-free dry particulate matter or tarr are presented4 as are.the effects of the filter structure on the delivery, of seie.ctedgas-phasec.omponents. 11
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THURSDAY MORNING - SESSION A -23- 8:40 Technical Session A: D. E. Blume, Presiding ALKALOID DEVELOPMENTINNORMAU AND CONVERTER'TORACCOS. R. F. Arrendkle,. R. F: Severson,., and 0.Ti. Chor~tyk, Tobacco SafetyRese.arch Unit,. USDA, SEA/AR„ Athens.,..GA30fi13, and G. R. Gwynn, Tobacco Research Laboratory, USDA, SEA/AR, Oxford, NC 27565. One of the most budworm-resistant tobaccos is aTobacco.Introduetion (TS-1112)i. This tobacco has also demonstGrate&a highdbgpee ofresi~stance to the green peach aphid. TiI-1112has the converter trait, resulting in high levels of nornicotine (about 75Xof the total alkaloids)) in the.cured leaf.. It was of interest to determine if thee insect resistance of TI-1i112could be aDtributed to a different alkaloid composition (nicotine, nornicotine, myosmine,, anabasine, anatabine) compared to alkaloid dis- tributions of conventional nonrr~esi'stant tobaecos.. TI-1.112and two normal bright tobaccos,.NC2326.and Speight G-28 „ were grown~at Oxford, NC, under flue-cured! . culturaliconditionss The tobaccos were sampled at the mid-stalk position at the following growth stages:: bud leaf, bud leaf develbpedito.fourto six inches, half-grown leaf,,full-grown leaf~,,leaf at timeofe toppdng,, ripe leaf before flue- curing, and leaf after, flue-curing. Green and ripe samples were freeze-driedimmediatelyaf'ter eollection, After extraction with KOH-methanol, the samples were analyzed by gas chromatography on a Superox-4 glass capillary.column,y usingg anitrogen~specific.detector. All tobaccos had similarr leveis of total alkaloids at the same stage of sampling. In TI-11i12, the majorityy of the conversion of nicotine tonornieotinetook.place after topping., The Leveisand distributions of the alkaloids at each stage of dpve.lbpment will be.di~scussed.. -24- 9:00 N1-I50PROPYLNORNICOTIPIE: ITS. FORMATIONFROM.NICOTINE.INAGED~LEAVES'OF NICOTIANA TABACUM. E. Lee.te:,.Depar2ment of Chemistry, Universi~tyof Minnesota.,Minneapoliis, MN 55455 More than 30 pyridine alkaloids have been isolated fzo-tobaceoleaves. Many of these compounds.are.the products of either enzymaticor chemical reactions which occur during the commercial curing and agingof'~ tobacco lleaves. Recently. Miyano and co-workers (Agric. BioZ.Chem;,, 43, 2205 (1979).)) reported on the isolation and: characterization of N''-isopropylnornicotinefrom air-cured burleyy tobacco. (Nicotianatabaeum).. We suspected that this compound', is formedfrom~nicotine. Aecordingly,, an aqueous solution of [2"-14C]Fnicot'1ne was painted on the leaves.of 4-month old tobacco.plants which were harvested three weeks later. This tracer was similarlyy applied to excised tobaccos leaves which were allowed todtyin air for four weeks. The alkaloids were extracted with~the addition of carrier N'-isopropylnornicotine. Radioactive nicotine and nornicotine wereisolated from theintact plants with onlyminuteacti'vityin the N'-isopropylnornicotine. All three of; these alkaloids were radioactive from,the air-curedi.leaves,.and degrad'ation of the labeled N'-isopropyl- nornicot'ine indicated that all the activity, was located in.the C-2" position. A higher level of activity was found in N'-isopropylnornicotine obtained from excised'leaves which were fed, .the[2'-1wC]niootine in aqueous acetone „ and'were treated on subsequent days with aqueous acetone. These results are consistent wi'~th the hypothesis that N'-isopropylnornicotine is produced in the curing,oftobacc.o Leaves byreaction~of, nornicotine (formed by the demethylatibnof, nicotine)wit}i~.aceto- aeetate,.followed by decarboxylati'on, and reduction. The 1:3C.NMR.chemieal shifftss of methyl groups ofN'-lisopropyDnornicotYne and related 1-isopropylpyrrolidines, which have.chLrality at then-position of thepy.rrolidine ring, are: significantly different (up to 7.5 ppm). 12
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THURSDAY ItIIORNING - SESSION'A -25- 9:20 DETERMINATION OF NICOTIVE,.vOR..NICOTINE,. AND OTHER SECONDARY AMINE ALKALOIDS INITOBACM B..M. Gordon and G..H. Greene, R. J.. Reynolds Tobacco Company, Research and Development, Winston-Salem, NC 27102. A,gas chromatographic (GG) method! has beem developed f~or the.quantitanive determioa- tion of nor~nicotine, anabaslne,, anatabine, myosmine, and nicotinee in,robac- The alkaloidkare extracted from~tobace-withaqueoussodium hydroxide, partitioned int~o chloroform, and filtered. Separation is performed utilizing: a 60~meter fused silica capillary column coated with 5E-54. Anethole is added as an internal standard with a flame ionization detector. Data are presented showing.near. 100'/..r~ecovery for nicotine and nornicotine from tobacco.samples. Aliso presentedlare data showing the percent relative standard deviation for the entire procedure for nicotine,, nor- nicotine, anabasine, myosmine~ and anatabihequantitation to be less than 3%. Inn a comparative study, thee percentt nicotine andthe pereent'total secondary alikaloids (asnornicotine)s were determined byy theCund3ff-Markunas A.O..A.C'~ t.itration method andithe newly developed GC procedure for over 400 different samples.. Good agreement'~ between the two methods was obtained when the values.for nornicotine.,, anabasine, myosmine,., and anatabine were summed in the GC procedure. In this same comparative study, a single sample was randomlyy inserted. Although the final values determined for total secondaryy alkaloids are essentiallyy identical, the coefficient of variance for the titration method is 25% and.for the CC methodd is 8a. -26- 9:40 CONFORMATIONAL ANALYSI6OF NICOTINE SALTS. T. A. Perfetti. R. Ji. Reynolds Tobacco Company, Research andiDevelbpmenD, Winston-Salem, NC 27102 The conformations of three types.of nicotinesalits have been determined. Thesesalits have acid:base ratios of either 1:1, 2:i1, or 3:11. Salt'formation between organic acids and nicotine is dependent upon.the.struc.ture of the acids (aliphat'~ic or aromatic) and their f~unctionality. The 1:1 salts of nicotinehave.aminoe acids or b'enzoic-type acid5bound to~the N-methyLpyrrolidihenitrogen of nicotine. The 2:1 salts are found to bind one acid group as in the caseoPe the 1:1 salts.and a second to the nitrogen of the pyridlne ring. The 2:1 saltsof, nicotinee are formed with~ formic acid, aliphaUic dicarboxylic.acidsand/ornitroaromatic acids. Nicotineforms. 3:1 salts with aLl aliphatic monocarboxylic acids from C'-2to C-M Here, one acidlis bound:as in thecase.of the 1:1 salts and the other two acids dimerize and bindlto~the nitrogen of the pyridine group. Iofrared „ ultraviolet, p.rotonmagnetic resonance, and.carb'on magneticc resonance (13CMR)) spectroscopy, as well asfielddesorptiom-massspectroscopy, were used ih this investigation of the conformation of nicotine.salts.. 13
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THURSDAY MORNING - SESSION A. -2q- 10:00 pD DEPENDENCEOF THE CARBON-13 NUCLEAR MAGNETIC RESONANCE SPECTRUM OF NICOTINE.. R. W: Slaven, Lorillard Corporation4 a Division of I Loews Theatres, Research Department, Creensboro „ NC'.274'20 The carbon-13 nmr spectra of avarietyofi nicotinium~ salts. haveb'een obtained at several acid:base ratios. Salts studied includptartrate, malate, and chloride. The spectra were measured at 8DMHzin deuterium oxide underr nitrogen using, freshly distilled nicotine. The chemical shi~ftsof the pyridyl carbons are found to be highly sensitive to the pD level.. Results are discussed both inn terms of thee pD dependeneeand the acid:base ratios these levelis represent. Graphical representations demonstrate that addition of acid greatly affects the closely spaced.resonances of the C-2 and C-6 carbons and the similaslyspaeed C-3 and C-4:carbons. At approximately pD = 8'uhere is.a crossover of the resonances in each pair. This observation is of special interest with regard to the assignments of C-2andiC-6.. The implications for previously, described chemical shift assignments ofthesec.arbons.are discussed. The influence of the chemical shifts of protonation.of the pyridyl andd pyrrotidy.ll nitrogens is also explored.. 10:20 INTERMISSION -28- 10:5'0 NEWCEMB'RANOIDSFROM BURLEY TOBACCO. V. Heemann~and U. Brummer,. BATi Cigaretten-FabrikenGmbH,.Forschungand. Entwicklung, Bahr,enfelder Chaussee.139,.D-2000 Hamburg 50, Germany Di2erpenoids are important precursors for tobaecof'lavor constituents.. To.gain more insight into the degradationmechanisms, the.distributionofditerpenoids in.aged burley tobacco was investigatedl For this aa liquid C02-extract of tobacco was separated by high-vacuum distillation and column chromatography using.sf~licage.l.A fraction enriched in dit~erpenoids eluted withdiet~hylieUier:. Individual.eompounds were isolated by repeated c.olUmn chromatography,.TLC, and HPLC on coiumns packed with Partisi1-10PAC., Ideneificationswere.aided byMS„ 'H and 1'3C NMR. All analysed diterpenoids had~the cembranoid structure. A new naturaliproduct„ the seco-lactone 4,8-dimethyl-8-hydroxy-ll-(1-methylethyl)-14-oxopenvadeca-5,9-di,en- 4-olide was identified „ and the structurewasensuredib'y ozonolysis. This compound could be derived from 2,6,11-cembratriene-4,8'-dioli byoxidativecleavageof the 11,12-double bond, a similar mechanism to that reported for the 4,6-diols. Three of the four possibleisomersof2,6,11-cembratriene-4,8-dioLs.were found. In addition to the well known a- and 6- 2",7,11-cembratriene-4,6-diols.and a- and B-8,11-epoxy-2,6 12(20)-cembratrien-4-ols, several other oembranoids new tob'urlieyo tobacco couldibe iflentified.. These results strongly support the.degradationschemes recently reported. 14
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THURSDAY MORNING - SESSION A \. -29- 111:10 FOUR.NEWCEMBRATRIENETRIOLSFROM TOBACCO. I. Wahlberg,I. Walliin, R. Arndt, K. Nordfors, T. Nishida, and C. R. Enzell, Swediish.TobaccoCompany, Research Department, P. 0; Box17001, „ S'-10462, Stockholm, Sweden Recent studieshavereveaLed that, dependent upon thegenetiebackground,tobacco varietiess synthesize cembranoids, labdanoids or both types of diterpenoids. These compounds are present in a fair amount in thecuticwlare wax of the green tobacco leaf, the (JS,2E,4R,6R,7E,11E).-and (iS,2E,4Ss6R_,7E,11E)-2,7',11-cembratriene-4,6- dio1 beimgthe major eembranoid andl(12Z)-ab'ienol the major labdanoid. We now report the isolation of four new cembranoids from a.Greektotiacco variety, which produces.both cembranoids and labdanoids. The new compounds have been formulated as the (1S',2E,4S,6R;7E,11S).-and (1Ss2E,4S,6R,7E,11R).-2~,7,12(20):-cembr~atriene-4,6,11- triols and the. (1S,2E,4S,6R,7E,10E,12S)-and (i1lS,2E,4S,6R,7E',10E,12R).-2,7,104cembra- triene-4,6s12-triols using.spectroscopic methods, particularl1y 1H and. "C NMR data, as well as chemical correlations with previously characterized cembr,anoids. The plausible biogenesis andib'iologieal role of; the new compounds will be discussed in the liigfit of results obtained bybiomdnoeticy synth'eses:. These have includedd sensi'tized photo-oxygenation of (1S,2E,4S,6R„7E,11E)-2,7,11-cembratriene-4,6-diol as well as model experiments Snvolvingenzyme preparations. -30- 11:30' 'CHEMICAL COMFOSITIONIOF DARK.AIR-CURED,AND FIRE-CUREDTOBACCO:. D.. L. Davis and B. H.. Song, Tobacco and Health Research Institute and Department of' A'gronomy.,, Univer.sityof Kentucky, Lexington „ KY40506, W. W., Weeks,. Crop Science Department, North~Ca:ro.lina State University, Raleigh, NC 27650 The objective ofthiss.tudy was to.compar~e the.chemical composition of dark tobacco cured byy two-ethods.and produced undbr three nit'rogenre.gimes. A Pargenumber of semi-volatile constituents were isolated from both types. Many of these were terpenoidswhichhave been identified as being.rel'ated to tobacco quality. Solanone andd several Cli3 ketones were identified. Solanoll, benzyll alcohol,. linalooliand'phenethyl alcohol were among,the compounds isolatedfrrom the st.eam.condensates. The phenolic compounds were present in higher concentrations.fr,om~ fr,om~tfire- cured tobacco. Anumber of different phenolic isomers were identified. One of the major compounds from dark fire-cured d'ist'illlateswas 4-Hydroxy-3'-methoxybenzoic acid. Several hydrocarbonss were identified in tobacco cured byy both~methods.. Nitrogen fertilization influenced:thee levels of certain nitrogenous.constituents as well as other compounds.;; however, otherss were unchanged by thesetr~eatments.. A limited examination of the influence.of s.toragewas conducted and the chemical, composition of these two types will be discussed'. 15 0
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THURSDAY MORNING - SESSION A -31- 11i: 50 CARBOHYDRATECONTENT'INBURLEY~ TOBACCO TISSUE. E. Leggett, J...Sims, and J.. Hamilton, USDA„ SEA/AR,. Department ofAgronomy„ Universityof. Kentucky.,.Lexington, KY 40546 Burley tobaccocv.. Kentucky 14 was grown on Spindl'etop~Farm.at Lexington,Kentucky. during.1~980. Thetotiaccowas produced using recommended cultural practices for burley, tobacc.oin Kentucky. Plants were harvested at topping and at maturity. These plantss weres.eparated into four stalk positions. Each stalk position~was separated into leaf lamina and stalk + stem. The samples werefreeze-drded and weighed. Analysis of t.issueinclud'ed total car~bohydtates,.reducing sugars.,.and starch~ The stalk + stem content of these components was greater than that of lamina. The lbwer stalk positions had higher levelsof, total carbohydrates, reducing sugars, and starch compared to the higher stalk.positions.. Total carbohydrates.in the lamina weree approximately 8%'.at topping and increased to a maxim-of 12% at maturity... Thee percent total carbohydrates in the stalk + stemdecreased from2b to 15 from lower mhigher stalkposition at topping, A'slight.deerease intotal.carbohy- drates occurred inn the lower stalk position at mat~uri'~ty. Reducing sugars decreased with an~increase in stalk position.. Thelamiharangedfrom 2.3 to 1.9%.in reducingg sugars whilie thee range for stalk.+. stem was 15 to 10%'from bottom to top~of' the plant. At maturity, the reducing sugars were increased in thelamina with essem- tiallyno change in the stalk + stem. Starch content was 77, and 1i0%in,.thelamina and stalk + stem, respectively,a.t topping.The lamina starch content was similar at maturity, but the starch decreased slightly in thes.taLk + stem~. Totali carbo- hydtates and starch content in toppedp,lants and un-topped plants were nearly equal at maturity. This observation may, be interpreted to mean photosynthesis decreased in both sets of plants and that seed productYon was not a good sink for carbon. 16
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THURSDAY MORNING - SESSION B Technical SessionlB: C. J. Rosene, Presiding A RAPID ME.THOD FOR:THE DETERMINATION OF N-NITROSONORNICOTINE INCIGARETTE.SHOKEBYGLASS CAPILLARY GAS CHROMATOGRAPHY. W. J. Chamberlain,, J. L. Baker, and R. F. Arrendale, Tobacco Safety ResearchUnit,.USDA„ SEA/AR,. Athens,. GA30613 • Arapdd glasscapiillary gas chromatography. (GO-2)) method for d'etermining.nitrosamine contents ih cigarette smoke condensate has been developed..Boththevolatdle nitrosamines and the nonvolatile N-nltrosonornicotine(NNN)were separated by GC on a fused! silica capillary column coated witH~Superox-4and detected with a nitrogen phosphorus detection system. Themettiod for thee preparation of these fused silica glass WCOT columns was developed in our laboratories and involves the use of Superox-4 as a surface pretreatment and deactivation agent and as a liquid phase.. These columns have produced excellent separatfonsof the nitrosamines in cigarette smoke. Smoke condensates from varioustypes.of'~ cigarettes weree analyzed, andl values of 0:0R to 1.77 ug NNN/cigarette were found. NNN levels in variouss tobacco types will be discussed. Thee method is offered as an.aliternative.forlaboratoriesnot equipped witHthe expensive.therma.li energy analyzer used in traditional nitrosamine analyses... -33'- 9:00: CONCURRENT GAS CHROMATOGRAPHIC DETERMINATION OF VOLATILE, NONVOLATILE,. AND TOBACCO~SPECIFIC N-NITROSA.MINES. K. D. Brunnemann andD: Hoffmann,.. American Health Foundation, Valhalla, NY10595 Tobaeco and tobacco smoke contain volatile nitrosamihes, the nonvolatile:V-niRroso- diethanolamine(NDELA), and the Cobaccos.pecific nitrosamines (TSNA).,.N'-nitroso- nornico.rine (NNN').,.4-(methyinitrosamino).-1-(3-pyridyl)-1-butanone (1NNK) and N'-nitrosoanatabine (NAT). For this investigation,, wedevelbped an analytical method which allows the concurrent determination of all three types ofnitrosamines., The smoke of 50 cigarettes was passed into two gas wash bottles containing ethyl acetate, ¢-tocopherol (as blocking agent), and 14C-14abeied dimethylnitrosamine, NNN, and! NDELA.as ihternal.standards. In the case of tobacco analysis, 25 g of tobacco were extracted with the same agents dissolved in ethyl acetabe.. Thec.oncentrated extract was.subjected too three chromatog.raphic.c.lean-upsteps during which thethr.ee.types ofnitrosamine.s were separated into a)', volatile nitrosamihes, b). NDELA, and c) TSNA.. The volatile nitrosamines weree analyzed by GC'-TEA'using Carbowax 2029 on GCQ at 200°C.. The NDELA fraction was evaporated to~dryness, redissolved in~ acetonitrile, and silylated1wit.h BSTFA. GC-TEA.analysis was performedl.on a OV-225 column operated at 140°'C.TheTSNAwere analysedd ditectlyy byGC'-TEA' on~UCW-98 at 185°C. The mainstream smoke of a commercialicigarette eonta.inedl 13,ngg dimethy.lnitrosamine, 11 ng nitrosopyrrolidine, 36 ng NDELA, 240 ng. NNN, , 110~ng NNK, and 330:ng. NAT_ A commerc.ial.fine-cut chewing tobaccovielded 11 ppb dimefhylnitro.samine, 93 ppb nifrosopyrrollidine, 740 ppb ni¢rosomorpholine„ 6,8 ppm.NDELA„ 31 ppm NNN, 31 ppm NAT, and 4.3 ppm NNK.(databased on~dryy weight)... Traces of nitrosoanabasiheweree alsoobserved.. The possible source for the newlyy discovered nitrosomorpholine will be di~scusse.d.. 17 0
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THURSDAY MORNING - SESSION B -34- 9:20 Among the components present in tobacco during homogenous.leaf curingis nitrate which has been associated with thef~ormation of nitroso eompoundk.and which may be removed by precipitation with nitron, a nucleophilic organic base. For thispurpose., nitron was complexedd with poly(vinylbenzyl chloride)) in the presence of iJ;,N-dimethyliformamide as;described by S. J. Chiou,and W. C. Danen. Tihis:polymer was.then coatedd on two inert supporting materials. (SephadexG-25„ coarse,, and acid-washedlsand).which were tested for nitrate capaeity, andiregenerative ability. , using,nitratestandards and tobaccoleafextracts.The.Sephadex.matrix adsorbed I 0.06 to 0.08 moles of nitrate per, mole of: nitronm.while the sand matrix retained 0.33 to 0.40 moles of nitrate per mole of nitron. Each support was capable of repeated regeneration with no detec.table loss.of nitrate capacity. Tobacco leafextracts.which eontaihed:0i3 to 1.4 umoles of: nitrate per gram of fresh weight.weret applied to~the Sephadex or sand matrices in.amounts.of0.12 or 0.46moles.ofnitrate per mole of nitron,.respectiveiy. In each case,thee polymerwas.abLe to adsorb nitrate with minimal interference from other components of homogenized tobacco. Thiss material also has the potential to remove other anions such as thiocyanate, chlorate, and nitrite. 1 -35- 9:40 APPLICATION OF A.SELECTIVE POLYMER FORREMOVAL OF NITRATE FROM TOBACCO. P.. R. Rhodes,,Department of. Agronomy.,,Universityo.fMaryiand „ College Park „ ND.20742and J... A. Saunders, Tobacco Research Laboratory, llSDA,; SEA/AR,: Beltsvil~le, MD 20705 APPLIGATION OF TENAX TRAPPING TO THE ANALYSIS OF GAS PHASE ORGANICS IN CIGARETTE SMOKE. C: E. Higgins,.W. H.. Griest, and G. Olerich, Oak Ridge National Laboratory, Analytical Chemistry. Divis.ion,.Oak Ri'dgey TN 37830 Capililarycolumn gas chromatographic analysis of the entiregasphasee of low-tar dpiivery.cigarettes.is not feasibleby, the usuali sampling methods. However, by smoking thec.igarette through a Tenax.trap placed directly behind aCambridke. filter,, theentire gas phase can be sampliedfor analysis by thermal desorption, andiprogrammed temperature.glass capillaryy gas.chromatography. The standard smoking parameters are used, and the puffprofilee is unaffected by, the Tenax trap. Breakthrough during trapping issignifieant only for high-tar deiivery cigarettes and.onlyfor constituents more volatile thanac.e.tone or acrolein. The application of more efficient.adsorbentsfor the collection of these volatile organicswiLl be discussed. For cigarettes of < 0'.2mg tar, per eigarettethe entireeontents of the trap are desorbedin the analysis. Traps used with higher tar delivery cigarettes are unloaded in a clean box, and apor.tionof' the well-mixed Tenax. or of a dilution made with clean Tenax, i'sanalyzedL Quantitation of identi.fiediconstituents is accamplishe.da by the use of appropriate exter:nal. standards added to Tenax andiana.lyzediby the.same method used for, cigarette smoke gas-phase constituents. External standard analyses wereneprod'ucible.to± 10%, and repro- ducib'ility.of cigarette smokeconstituente analyses averaged ± 17, 22, and 502for cigarettes.of high(36 mg), Iow (7-19 mg)~, and ultra-1ow (0',01 mg) tar, deliiveries „ respectively. This precision.mayn be improved through,theuseofe an internal' standard. The deliveries of"-30 representative gas-phase constit- uents.ranging in~voliatility from isoprene to Limonene arepresented for cigaretteswith tar deliveries ranging from 52 to 0.01 mg., 18
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THURSDAY MORNING - SESSION B -36- 10:00 I ISOLATION AND LDENTIFICATION',OF TWONEWSECO-CEMBRANOLDS FROM, GIGARETfrESM@KE.. T. Katz,,R'. N. Ferguson„ R, D.Kinser~, and!W: N., Einolf,Philip Morris Research Center, Richmond, VA232b11 and T. P. Pitner; Boehringer Ingelheim„ Ltd., Ridkef;ield, CT 06877 We wish to.report the isolation andlidentifieation of two new.seco-cembr~anoids, (5S.): 5-isopropy1,8,12-dimethyl-3E,SE,12E,1'4-pentadecatetlraen-2'-one andit.s 12Z' isomerfromc.igarettesmoke condensate. These new C2p:acyclicterpenoids were isolated from smoke eondensateby partitioningtietween methanol/water and cyclo- hexane followed byy partitioning of the cyclohexane residue between cyclohexane andinitromethane.. Next, silicage1 column chromatog:raphy,.f'ollowed byy preparativeglr,.was used to isolatemill'Sgram~quantitiesof each isomer. Spectroscopy(IR,. MS, HNMR, and 13C NMR)iwas used to,identify these compounds. Also, double bond positions were located by ozonolysis of thealL trans-seco-cembranoid followed byy treatment of the reaction mdxture~withtriphenyiphosphine. Finally, thermal treatment o.fa- or B-4',8,13-duvatriene-li,3-diol was shown to.producethe neww seco-cembranoids which supports the proposedlst'xuctures. Previously, acid catalyzediringopening of °-4,8,.1i3-duvatriene-1,3-dio1 was shown to produce the related seco-aldehyde, 3„7; 13-trimethyl-l0-isopropyl-2;6.,1Q,13-t'etradpcatetraen- 1-al,, a member, of a classs of CZpterpenoids which is hypothesizedd tobea precursor of numerous volatile tobacco flavor compound's-. Therefor.e„ webeli'evethat thesenewseco-cembranoidsf'ormed1by thermal ring opening of duvatriene-1,3- diols mayy offer other precursors to this class of tobaccofSavorants., 1'0:20' INTERMISSION -37- 10:50 SL"SULTANEOUS, RAPID GAS CHROMATOGRAPHIC DETERMINATION! OF WATER AND NICOTINE IN SMOKE. B. M. Gordon, R. J. Reynolds Tobacco Company,, Research and'Development.,,Winst'on-Sa1em~ NC 2710.2 An improved method for the simultaneous determination of nicotine and water in smoke. TPM has been developed. Filter discs are transferred:to a flask containing a suitable solvent and extracted for 30 minutes on a mechanical shaker. Theanaly:ses are.accomplished.with Hewlett-Packard Model 5880A' gas chromatographs equipped with both flame ionization (FID): and thermal conductivityy detectors (TCD)... AchromatographicA analysis of nicotine performed on a newly.developed' column packing.material prepared by. Whatman,,Inc. is.descr.ibed'. The.combination of detectors and automatic sampling characteristics of theHP-5880A-7672A.aLlowsthe loading of up to 99'samples. The auoo-iinjectorsampies a single vial and, at timed intervals,, injects pre-selected voliumesonto each of the.twocolumns required for nicotine and water quantitatfon.. When all peaks havee7utedfrom, the column, thenext sample is injected. This continues up too the99.-sample limit. Thee analysiss time for the nicotine~determination has been reduced from six minutes to approximately threeminutesutiliizing.the Whatman column, whilee yielding baseline resolution between the nicotine peak andithe internali standard peak. Precision data for the quantitation of nicotine are presented with anetholee or hexadecane as internal.standard!. 19
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I THURSDAY MORNING - SESSION B -38- A RAPID GAS CHROMATOGRAPHIC DETERMINATION.OF MENTHOL.INTOBACCO... 11•10 B. M. Gordon and M. F. Bargerding, R. J. ReynoldGTobacco Company, Research and Development, Winston-SaLem, NC 27102' A rapid gas chromatographic metho&for the determination of inentihol:intobacco was developed. The method was designed to be simple andito provide high sample through- put with good aeeuracy and precision. The method includes asteam~distilllation,ofi tobacco with the menthol dis.tililatecoLlected in a final solution of 50/5:0. (v/v) isopropyl alicohol/water. Anethole is added as an internal standard. Gas chromata- graphicanalysisis performed on an HP-5880A gas chromatograpM with dual 1V8 in. stainless steel columns. The columns are packed with 3Y'OV-101 on a Chromasorb W solid'support. A short discussion of the characteristics unique to a column used in fast analyses and the techniques used to prepare these eolumns isinclhded. The HP-7672A automatic sampler in combination with the KEYSTROKE PROGRAMMING capability of the 5880A GC enables sequential sample injections on both columns in the gas chromatograpH6 This essentially doubles'thee sample throughput/GC/hr. The.internal standard method is used for sample quantitation. A.CI6-CALS. computer, system is used f~or data reduction. Sample concentrations in a rangeo.f 0.2X:to li.5Xtiy weight are determined. The relative standard deviationfor the procedure isless than 3%. -39- 11i; 30 LOW MOLECULAR WEIGHT FATTY ACID~SUGARESTERS INITURKISHiTOBACCO: SEPARATION BY REVERBE-PHASEHIGH PERFORMANCELIQU.ID. CHROMATOGRAPHY AND SPECTRAL CHARACTERIZATION. J...M. Rivers, R. J.. Reynolds Tobacco Company.,. Research and Development,Winst'on-Salem, NC 27102 Four homologous mixed sugar estersofCz., C5, and C6 acids with masses 474, 488, 502, and 516 have been~isolated from Turkish tobacco:., A sample of Turkishitobaceo:Smyrna (Izmdi)AG, 1975.crop,,was.extracted with hexane,.and a crude fraction containing sugar esters wasobtaihedby variouss partitioning steps., The sugar esters were enriched by normal-phase high performance liquid cHromatography, (HPLC)andseparatedi by reverse-phaseHPLC. Characterization by glass capililarygas chromatography, field d'esorption~mass speetrometry, carbon~and proton magnetic resonance,.and infrared spectroscopyy indicated that the four isolated sugar esters with masses 474',,488, 502, and 516 were tetraesters with at least one acetate group. For the three remaining ester moiet'ies,.the sugar esters with masses 474, 488, and1502had different combinations of isovalerate, 2-methylbutyrate, and 3'-methylvaler.ate groups, while the sugar ester with, mass 516 had only 3-methylvalerate groups. The sugar ester with'mass 516 had been previouslyy i'soiat~ed and characterized as. 6-0-aaetyl-2,3,4-tri'-0-[(+):-3-methy:lvaleryl]-6-D-glucopyrano.se. 20 ®
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THURSDAY MORNING - SESSION B' -40- 11:50 DEVELOPMENTi OF METHODOLOGY FOR THE DETERKINATQON OF'BROMIDERESIDUES I5TOBACCO ANDTOBACCO.SMOKE. FACTORS AFFECTiINGTHE RESIDUE LEVELS RESULTING FROM~METHYL BROMIDE FUMIGATION'. W. Qartwright, A. Cornell,, and V. Olender,. Consolidated,Cigar Company,.131 Oak Street,. Glastonbury, CTi 06033 J Methyl bromide fumigation is inc.reasinglyy being,used to control tobacco beetle populations. The obje.ctivesof~ this work were: 11).thedevelopment) of rapid analytical procedbres which could be used routinely for the d'eterminationn off residual bromi'destemm~irtgfrom methyl bromide fumigation;~and 2)) assessment of f:actorswhich influence thee levels of bromide residue arising from methy:llbromidee fumigation. Comparison~was madeamong three procedures: 1)) oxygen~tiomb;. 2)~titr.imetric; and 3)) specific ion electrode. The specific ion electrode procedure was adopted for routine use and provided results comparable to the highly.aceurate „ but expensive, neutran activation analysis. Neutron activation was, however,: used'to semi-quantitatively assess mainstream..and sidestream concentratdonsof bromide. The residuaL bromide: wasinfLuenced by.several factors. Finished ci~gars, ass well as component parts (wrapper/bihder/.f,ill'er), were fumigatedland analyzed.. Residual bromide increased with increasing: methyl bromide dosage or duration of fumigationibut wasreducedlb'y multiple air-washing. Both the type of tobacco and the tobaccomofsture content had a limited influence om residual bromidelevel's. The effect of multiple fumigations was alsoinvesti- gat'.ed. Pertdnent dat'.aa will be presented and'discussed for the above topics.. 21;
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THURSDAY AFTERNOON - SESSION A 0 -1 pJ Technical Session A: L. D: Story, Presiding -41- 1i:40 DETERMINATION OF M@LYBDENUMINBURLEY TOBACCO USING A.RAPID AU'IOMATEDI METHOD. F. Eiyazi, J. L. Sims, and J..Crutchfieldl. Department of Agronomy, Universityy of Kentucky, KY 40546 Anumb'e.r.of methods have been proposed f~or.theroutihedetermination ofMo,in plant materials. However, most.enemical methods previously described involved solventt extraction in whi'ch both tfieaccuracyand precision of the method deteriorate at levels approaching Oi1 ppm Mo,, the critical levelifor many piants.. Withthe thiocyanate spectrophotometric proeedure,.diff'~iculties have been encountered with interferences from Fe, the use of stannous chloride as a reducing agent, and isoamyl alcohol saturatedlwith carbon tetrachlbridef~ortheex2raction of the.c.olored complex. The useof'e procedures such as directt atomic absorption and'spectrographic metlhods is restricted by poor detection limits.causeds by the refractory nature of Mo oxides. A method is described f'~or Mo determination based on itscatalytieaction on the potassium iodide-hydr~ogen peroxide reac[ionL An autoanaly.zer with a modified de-bubblingsystem.and aa longpath length (50 mm)) flow.cell is used. The procedure is sensitive enough to determine <Oll ppm of Mo in plant material, usinga.0.50.gplant sample. The method has.been applied f~or direct determination of the element in burley tobaeco,, soybean, and'~orchard plant samples after dry ashing~ Interferencesfrom.high concentmations.of~ certain salts (>5Y.), have been encountered but can be overcome by.addition of appropriate amounts of salt to the standard solutions as their chlorides. -42- VARIATION INMINERAL.CONTENT OF FLUE-CURED TOBACCOS DURING CURING. 2:00 j A. I. Schepartz,.Tobacco.Safety Research Unit, and R. L. Wilson, Biometrical Services, USDA, SEA/AR, Athens,.CA,3061i3, and J. F. Chaplin,. Tobacco Research Laboratory, USDA, SEA(AR,.Oxford,. NC 27565. An analytical studywasy made of the levels of.mineralis present in two fiiue-cured tobacco varieties, NC:955 and;LA 53(lownicotine).,. during,five ihtervaLss in the curing process:: at harvest, half-way through yellowing, end of yellowing, end of' leafidrying, and completelycur~ed. The tobaccos were grown at Oxford, NC, during the 1975 crop year and sampled according to arandomiied square design.. Leaves were harvested in four primings with three replicates of each priming.. Curing wass conducted bythey conventional barn methodl Thi~rteen.el'ementsweredetermined on each sample: phosphorus, potassium,, caleium,magnesium,, manganese,., iron, aluminum, boron, copper, zinc „ strontium.,,barium, andisodium. Res.ults.wereevaluated statisticalily via computer,, providing an analysis of variance and graphic plots from~which trends could be predicted. Although these elements wouldlordinarilybe regardedd as stablee andinonvolatile, significant decreases and increases were noted for several of them. Leaves.of the firstt priming and, in some eases,, the fourth priming appeared to be.quite different from~ the other.r primings ih several ways. 22'
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THURSDAY AFTERNOON - SESSION A -43- 2:20 I THE CUTICULAR.CHEMISTRYOF GREEN TOBACCO LEAF. R. F. Severson, R. F. Arrendale, 01 Ti. Chortyk, and L. B. Smith, Tobacco Safety Research Unit, USDA, SEA/AR, Athens, GA30613„ J. F. Chaplin and G. R.Gwynn, Tobacco ROsearch Laboratory.,, USDA, SEA'/AR', Oxford, NC 27565, and A. W. Johnson,.Pee Dee Experiment Stationy Flbrence, SC 29503 Recent workNas indicated that the eomposition.of the green leaf'f cuticular terpenes mayb'e of significant importance. It has been reported that the degradation during curing and'.aging of cycliic terpenes results in the production of numerous volatile compounds that may also be important tobacco flavor constituents, that cuticular labdenoid5.and cembrenoid's of'~ tobacco have plant growth inh'ibitingproperties, and that the surface ehemicalsmay.be responsible.for insect and blue-mold resist~ance in some tobacco type.s... These observations indicate that the green~tobacco leaf surface compound'smay play an important role in develbping.low tar, pest-resistant high flavor tobaccos. We have continuediour studies on the composition of the cuticular waxes. Younggr~eenleaves were washed with methylene chloride to remove leaf surface comp,onentsand the washes were subjected to~chromatographiesepara- tions and analyses. The.nonpolar components were identified asC2,5-C36 paraffinic hydrocarbons and wax esters (C32-C44). Thepredom~inant aLcohoisof these wax esters were.eosa:nol and docosanol, while the acid moieties consisted of C14, C15, C16, and C18 saturated fatty acids. The medium~polarity, compounds were primarily free paraffinic alcohols.Ttie major components on most tobacco types were the diterpenes. Flue-cured varieties produce thecembrenoids„ a- and B-dhvat.rienediols. Some tobaccos produce labdane compounds (abieno1andre.lated'campounds).. Other tobaccos have both labdanes and cembranes. Af~ew tobaccos also produce polareste.rs of low molecular weight acidswi~th sucrose. -44- 2:40 THEQ@ANTITATIONOF THE MAJOR'CUT'ICULAR LEAF COMPONENTS OF GREENTOBACCOL . R. F:Severson„ R, F. Arrendale, 0. T. Chortyk,. K. L. McDuffie, and ~ P. F.. Mason, Tobaeco,SafetyResearch.Unit., USDA, SEA/AR, Athens, GA30613,. A. W., JoAnson,. Fee Dee Experiment Station, Florence, SC. 29503y M. D. Jackson, G: R. Gwynn,.andlJ. F..Chap,lin,.Tob'acco ResearchLab'oratory, USDA„ SEA/AR,. Oxford, NC 27565 The role that surface components of green leaf may playin host-plant resistanceande in the development of flavor of cured tobacco prompted us to develop a rapid method for their quantitation. Green leaf was collected in the fiel&and was dipped in-thylene chloride.toe wash',of'f tfiesurfaeecomponents. In the laboratory, concentrated wash, from about 0.3 to 0.6~g of green weight, was added to.ano internal standard solution of heptadecanol in a.taperedtest tube. The solvent was removedwith,a stream of nitrogen and 50 u1 of 1~:1 N',0-b-is([rf- methylsily.ll)trifluoroaeetamide.in~dimethyi f~ormamid.e was added., Thet'est.tutie was capped andlkeated at 7.5°C for 30 min. After cooling, a 50-u1 portion of 1:11 i7,0-oi,.(trimethylsi~lyl)ac.etamidein pyridinee was.added top.eevent precipitation of the hydrocarbons. The mixture was transferred to a micro autosampller vial and~ analyzed by gas chromatography on a 3D.m:SE-54 capi~llarycolumn. Quantitative data were obtained for the major di2erpenoids, h'ydrocarbons „ docosanol, andpolarhigh molpcular weight esters. The gaschromatographic- p,rof2~les.and thecalcul'ated levels of cuticular componentsf'~rom varioustotiaccos will be discussed. 3:0& INTERMISSION 23
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-45- 3:30~ THURSDAY AFTERNOON - SESSION A THE FRACTIONATIONIAND ISOLATION OF THE CUTICULARCO'.HPONENTSOF GREEN! TOBACCO. R. F. Severson, 0..Ti. Chor.tyk, and K. L. McDuffie, TobaccoSafety Research Unit,,USDA, SEA/AR, Athens, GA' 30613, A. W. Johnson, Pee Dee Experiment Station,. Florence,.SC 29:503, M. D.. Jackson, C. R. Gwynn, and J. F. Chaplin, T~obacooResearch Laboratory.,.USDA, SEA/AR, Oxford, NC 27565 To~investigdte theroPe that green leaf cuticular components play in host-plant resistance, it was necessary to separate the various classes of compounds (hydrocarb'ons.,.wax esters, paraffinicc alcohols,.diter~penes and related components,, and the polar esters). Wefoundithate a simple solvent.partit'doningstep,..using hexane and 80%MeOH-H20, separated the.surface chemicals into polar and nonpolar fractions. Thehydrocarbo.ns„ wax.e.sters, paraffinic alcohols, and'cis-abienol were concentrated in the hexane fraction4 while the duvatrienediols, 13,E-labda- 13-ene-8a,15-d¢od (a 15.hydroxabdenoli)', and polar e.sters.were concentratedl.in the methanol-waterfraetfon.. Paraffinic hydrocarbon„ wax ester, and paraffinic alicohol fractions,., as well as individual diterpenes,.can~b.e obtained by chroma- tography on alumina. The polar esters were isolated bychromatography.onSephadex LH-20with a:chloroform-methanol solvent system. The ch'romatograp.hic conditions and recrystallization systems usedlto,obtainditerpenes as well as the tobacco types used in theisolation ofeaeh~type of component will be discussed. -4'6- 3:50 COMPARISON OF THE SURFACE CHARACTERISTICS AND CHEMISTRY OFNOR"1P.L AND CREENIPEACH APHID RESTSTANT.TOBACCOS. R. F. Severson and K. L. McDuffde,.Tob'acco Safety. Research Unit, USDA, SEAIAR,.Athens„ GA 30613 and A. W. Johnson,.Pee Dee Experiment Station, Florence, SC 29503 In reeent years, increased green~peach aphid levels on tobacco resulted in corresponding greater use of insecti~cid'es. However, a more desirable approach for aphid control would be the development of commercially-acceptab'le,, aphid-resistant tobaccos. Suehir,esistance has been reported inNicotiar.ay but the nature of the.observed resistance is not understood.. It has been speculated that surface trichomes anditheir secretions may be responsible forresisGance. In thisp,resentaoion„ we willireport, our initial studies to determine both chemical and physical leaf',surfac.ee differences between aphid- resistant and susceptible tobaccos. During 1980, 24.resistant tobaccos and two,susceptible flue-cured vardetieswere grown at the Pee Dee Eicperimen0 Station, Florence, SC., Visual observation of leaf trichome characteristics and their exudate.s were.made. Young green leaves were qPanUitatively washed with metlhyl'enechlorid'e:to remove cuticular~ chemicals.. Thee chemical constituents of the washes were.characterized by our glass capillary GC profile method.. The relatfonshipsb:etween~leaf surface chemicals„ trichomecharacteristics, and aphid resistance will be discussed., 4:30' BUSINESS MEETING 24 j G
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THURSDAY AFTERNOON - SESSION B Technical Session B: E. Glock, Presiding -47- I THERMAL DECOMPOSITION OF PECTIN. K. R. 8quireand B. E: Waymack. 1:40 PhilipMorris Research~Center, P. 0. Box 26553,Riehmond,.VA23261 To.understand tobacco carbohydrate pyrolysis, the thermal decomposition of citrus pectin was stiudiediusing evolved gas analysis by gas chromatography. Both a TC detector with a Carb'osieve B column and a FID,deteetorwitha Porapak.Q column were used too simultaneously analyze for H2, CO, C02,,, water, methanoL,.methyl'formate.,, furan, methylf;uran, dimethyifuran,.fur~fural, C1-C4.alkanes, C2-C4 alkenes,.C1-C5 aidehydes, C3-C4ketones,,and C1~C3'.carboxylic acids. The sampLes werepyrolyzed in flowingHe.at 1°O/min. The majprdecomposition products were C02,(4'57mg/g).,. H20 (286.mg/g).,, CH30HI(83 mg/g).,. CO (455 mg/g), CH4.(11 mg/g), and H2(6'. mg/g). TheC02. is attributed to deearboxylation of the C1 carbon and the cartioxylieacid side gr~oup of the.galacturonic aciH-ings. Water isevolNeditioth from~bound water and1b'y dehydtationof the galacturonic acitlringsd giving approximately one molecul!e of water per moleculeo.f,C02. Methanol is a product off hydrolysis and ester exchange reactions involvi'ngthe galacGuronic acid methyl esters. CO is a by-product of reactions leading to other carbonyl compounds. CH4and!HZ are formed during.Ghermal cracking off thec.arbonized char left after the initial major decompositions. Analysis of the homologous seriess shows that the ord'eringof peakevolutio-temperatur.esk bychaih length for the hydrocarbons is oppositee of those observed fortheald'e}iyde„ ketone,., acid, andlfuran series andlindicatesthat thermal erackingcompeteswith char fragvientation. -4'8- 2:00 THE EFFECT OF SOME '.JITROGENOUSBLEND COMPONENTS ONNO/NO* AND HCN LEVELS IN MAINSTREAM AND SIDESTREAM SMOKE.V'. Norman, A. M. Ihrig,. T. M. Larson and B. L. Moss, Lorillard Research Center, Greensboro, NC 27420 The relationshipsbe.tween nitrate and total vollatilebase contents of t'~he tobacco blend and the NO/NO% yi'elds.ofs mainstream and sidestream smoke were examined. The NO concentration of the vapor phase waslinearlyreLated'to the blend nitrate leve.loveral rangef'som,. 0.07Z to 3.70%.. When extrapolated to;OYblend nitrate level there remained a residual NO concentration which apparent4yy arises from precursors,other than nitrate: Native nitrate from flue-cured/burley blends and added~nitrate in the for-ofMg(N03)".2•6H2O~ at comparable levels, gave similar NO.concentrat'ions. HCN coneenf~ration-as not appr.eciably affected by the bLend'nit'rate level.. The total volatile base content of the bliend:had little effect on either the HCN or the NO/NOX yields. Data will be presentediconcerningg the effect ofblend.nit.rate level on the sidestreamto.mainstream.ratio.of NOO and on the presence of NO.g/ti02 in the particulate phase. 25
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THURSDAY AFTERNOON - SESSION'B' -49- 2:20 FORMATION AND TRANSFER OF 4-(METHYLN'ITROSAMIiNO)-1-(3-FYRIDYL)-BUTANO~E (NNK)DURING SMOKING.J. D.Ad'ams,A. Castonguay, S. J. Lee, N. VinchkoskY, and1D. Hoffmann, American Health Foundation; Valhalla, NY 10595 N'-Nitrosonornicotine (NNN) and NNK are tobacco specific N-nitrosamines (TSNA)iwhdch are formed during tobacco processing as well as d'uring smoking. Earliier, we havesh'own that some of the NNN',in processed cigarettetobaceo, transfers unchanged into smoke and that the remainingNNN is formed duringsmoking(55Y,). Whereas NNN originatesfrom nicotine and nornicotine, NNKcanonlybe derived fromnicotineby nitrosation. Todetermine theordgih of NNKinthesmoke,wespiked spikedcigarett NNK-1-140 for the assay of the transfer and with nicotine-methyl-74C for calculating the extent of the pyrosynthesis from nicotine. T'hemainstreamsmoke(MS)was trapped in buffersolutlon (pH: 4.5)1 containing ascorbic acid!as blocking agent and NNKr3H as an internali standard. The organic extract wascolumnchromatographed „ the NNK-metfiyl-''b fraction wasisolated~b'y HPLC and analyzed byGC-TEA. In~the case of a UL S. nonfilter cigarette (85 mm), we found that 7% of the NNKin~the tobacco transfers unchanged into MSS whereas0.DD8Y.of the nicotine in the tobacco column gives risetoNNK. Of the NNK, 37%is transferred~and 63% pyrosynthesized,fr~om nicotlineduring stmoking. Wewilil discuss these results and'the fate of nicotineand NNRduringsmokingof cigaretltes deliveringweaklyacidic and weakly alkaline smoke as well as data onthe reduction~of~ NNKand other TSNAin cigarette smoke. -50- 2:40 PRECURSORSTUDIESON CATECHOL IN CIGARETTESMOKE. S. Carmella, S. S. Hecht, and D: Hoffmann„ American Health Foundation, Valhalla, NY 10595 Our earlier experiments indicated that possible precursors to catechol in cigarette smoke included sugars, polyphenols, celilulose, andlignin. However, studies with 14C-labelled sugars showed that these were not major precursors. Therefore, the contribution of polyphenols, cellulose, and lignin~t~o smoke catechol have been investigd.ted.Levels of chlorogenic acid and rutin~in 1R1 tobacco were determined by highpressure liquid chromatognaphy. Tobaccowas sonicallydhsperse&with water and the extract was analyzed byreverse phasechromatographyon a WhatmanODS-2column employing a solvent program as follows: 90% solvent Ato~100%solvent B in 401mmn at 1 mli/min, Solvent A was0',05 M KH2P04adyusted to pH 2,9and solvent B was 80%methanol/water.Cfilorogenic acid and rutin eluted at 21.8 min and 31.5 min, respectdvely; and were detectedlb'y UVabsorbanceaC 340 nm, Levels of chlorogenic acid and rutin in 1R1 tobaccowere 7.6-nd 4,.1mg/gram dry weight tobacco, respectively. Toinvestigatethe role ofohlorogenic acids groups of 1R1 cigarettes were spiked with 0 mg, 4mgy 7 mg,10mg„ or14tmg by the syringe technique. The smoke of 10 cigarettes from each group wasthen analyzed for catechol. Catechol levels in the smoke of cigarettes spiked with chlorogenic acid vere:control1R1, 320Lg/cig;4 mg chlbrogenic acid added, 320 ug/cig; 7mg 400ug/cig; 10:mg, 360:ug/cig; 14mg, 330 ug/eig, The resultsunder these experimental eondi2lonssuggest thatctilorogenic acid is not a major precursor of eaoechoi incigarette smoke. The levels of catechol in cigarettes treated with~r,utih, cellulose, orlignin will also be reportedL 3:00 INTERMISSION 26 I
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THURSDAYAFTiERNOON - SESSION B CIGARETTE PACK MOISTUREVARIATSON INSTA.NDARD 9HIPPSNG CASES. K. A. Flaherty, Btown and Williamson Tobacco Corporationy tioui'sville,. KY 40232 Variousstudies~were conducted to~identify cigarette pack moisture content by, carton,, pack and layer posi~tions in standard shipping ease.s.. The primary objective was to determine the moisture profile and degree ofvariatiom within the cases in commercial storage. Initiai environment cabinet testing (100PF,.20X RH).wasconducted with 6M cases. A one-year Phoenix Arizona warehouse study traekedthe moisture content of ninety unique paek.positions in 12M.cases. A similar one-year study in a Louisville, Kentucky warehouse analyzed the pack positions of four selecoed carton positions in both 6M and 1'2M cases. Oigarettetobaccowas analyzed by the standard tobaceo,oven moisture method. Results indicate a misturegradient developsin a case with age. Wider variation in stored product moisture content occurs with inereasingage. Differences in moisburecont~ent levels were identified'byy carton, pack.and layer position over the test periodk. -52- 3i50 AMODIPIEDQUALIT'YINDEX OF FRESH CIGARETTE.SMOKE. J.-C. Park, ' T.-H. Lee, and K'.-K.Yoo. Korea Ginseng and T~obaceo,Research Institute.,. Seoul, Korea 110i Korean non-blended flue-cured cigarettes and Korean.commercial cigarettes were examined for evaluatloTn ofsmokequal'ity. The SH~indexof each eigarettewas obtained for the amount of cysteine.thiol groups which reacted with such,.smoke components as unsaturated carbonyls, nitzogenoxides, and activated double-bonded compounds. This index wascombined'with the measured amounts of tar (T).,nieotine (,N)~, and carbon monoxide (C)in the smoke to provide the indices SH((N+T)iand SH/(N+T+C). For cigarettes with deliveriesof.tar, nicotine,, an&carbon monoxide of 9, 0.6.and 8 mg per clgarette,respectdvely, these indices become SH/.[N/0.6+ (T+C),/8] and SH/(NY0.6 + T/8)_ The correlation coefficients (r)tietween the.sei'ndices and stalk positions of Korean non-blended flue-cur~ed cigarettes were -D:5766 for SH and'-0.6044for SH/(N10.6 + T/S)„ the latter being,the better index. Fbr thecommercial cigarett~es,.ttie correlation coefficients between the ind.ividual indices and the cigarette grades were 0.5789 for T/N,-0.6403forSH/[N/0.6+ (T+C)/8], -0.6623 for SH9(N/0.6+ T,/8),, and. -0.7080 for SHJ(N+T), indicating the latter is the most reliable index. From theresultsobtained,e carbm.monoxidedoes not s.eem~toc.ontribute significantlyta smoke quality. 4:30 BUSINESS MEETING. 27
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FRIDAY MORNI!NG Technical Sessiom D. L. Roberts, Presiding 8:45 PH'ILIP MORRIS'AWARDEE, RESEARCH HIGHLIGHTS -53- 9:15 r A.METHOD~FOR CONSTRUCTING~WEIGHT DISTRIBU'TIONS VSSHRED DIMENSIONS. M, E. Counts, and B. C. LaRoy, Philip Morris Research Center~, Richmond, VA 23261 A method iss described for transforming shred dimension fcequencydistributions into corre.sponding,weight distributions by image analysis. The ability.of the computerized image analyzer to quantitate shred dimensions has numerous applicationstos tobacco research and processing, but knowledge of how weight is distributed among shred dimensions is frequentlyy required. Sieving provides weight distributions which are complex functions of density, an,lgeometry. Thus, thee objective of this work was number-to-weight conversfon~of'f image analysis data. The method described was.applied.oo one typee of tobacco filler before and after makerr handling..Slired dimensions and image analyzed sample weights were used to derivee the weightt distributions wfthin~individual sieve fractions. Fromthe sieve fraction weights, the component sieve fractions were recombined:to obtain weight distributions vsshred length and width~£o.r thecotal sample. These were satisfactorily described by 1og-normal curves. Maker handling resulitedlin a narrowing of the weight. as length, distribution primarily from the high-length side of the curve with~little change.in maximum and minimum shred length. Numerical inte.gratfon.ofn the fitted total sample dfstsibution.curves was done to obtain included weight f.ractionss equal tosi~evefractions. "Effective sieve lengths" from theseintegrationse were similar for the two samples despite ttieir different weight distributions. The method may.be.applicab'ley to studies of other tobacco fillers. -54- 9:35 A METHOD FOR MEASURING THE AREA OF TIPPING PERFORATSONS.. P. F. Perfetti,. Research and'Development,. R. J. Tobacco Company, Winston-Salemt NC 27102 A.method for measuring the area of electrostatic, laser, or.mechanical tippi~ngperforations.wae deve~ qed. A photomicrograph~of anindividual perforation was taken with~a Leitz.ORTHOpIAlY light microscope equippedw4~th a Poliaroid camera. Coordinate data for the center point andithe points on the perimeter of the perforation imagee were collected with a.Tektronix 49566 graphics tablet interfaced to a Tektronix 4052 graphic computing system. The area of the perforation was determined bysummingthey areas of a1ltriangLes that are formed by the centerpoint and two adjacent perimeter, points. The area of each triangle was calculated with thefolilowing equation: Area - 0.5 (xlyZ - xZyi + x2y3.- x3y2.+ x3y1i - x1y3): where the vertices of' the triangle wete (x~,yl)', (x~, y~and (x3, y3). The actual perforation area was then ealculated with he magni icfl ion of the phonograph. Fifty perforations were measured intrl~pliicate. The coefficient of variationwas calculatedlfor each.perforation.. The average.coeffieient of variation, a measure of the precision of the method, was found too be between 1.5 and2.2X.Tlie error of thee method was found tobeo between 1.2 and 1.6%. This range was determined by, comparing theareavalue determined by this method with the area value calculated fr,om,the diameter of two apertures of known diameter. 28'
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FRIDAY MORNING -55- CALIBRATION AtYD~CORRELATI09 OFTSiO'TRPES'OFCORESTA.P£RMFABILIT~YMETERS. 9:55 j A. L. Benson~, T: Kenefick,R, McCarthy, and P. Suoherl'and'y C. H. Dexter Division, 2'Elm Streer, Windsor Locks, CT'06096 Tlhe objective of this study. was to prepare two types of' CORESTA permeability meters for routine, reliable use in product development andd quality control of! porous plug wrap, One met~erwas designed by Dexter and measures mass flow with~a hot wire anemometer. The secondd meter was purchased from Phobos, Inc. and~me.asures volu- metric flow with two~Merriam laminar flowmeters. Bothmet'.ers test 1-cmZareas. Meter ranges are 600ito 60,000 cm~/min/centibar (Dexter) and 5 to.50,000 em/min/' centibar (Phobos). Calibration standards were a Merriam,lam,inarflow meter and an inclined manometer. The pressure drop of sample holders wasnegl!igitile up to20,0D0'cm/min, increasing,oo about 2mmHz0at 40,000:cm/min. Correlations were obtained with 30~orifieeplates and 30 porous plug wrap samples over the range from 900to 44',000cm/m~ia/cent'ibar. Plug wrap-amples wer.ee tested two.ways: same test areas and pairedd samples. Standard parametaicand non-parametric correlational statistical analysistechniques were.used. Destructive effects on plug wrap~ofl repeated'~testing of ohe same area were considered. The effect of ambient pressure on meter accuracywasal'so considered. A need'forstandard calibration and correlation methods isstated,s and recommendations are made for further work.. -56- I COMPUTER STUDIES ON THERELATIONSHDP OF FLAVOR TOyOLECULAR SHAPE. 10:15 R. S. Marmor, Lorillard. Corporation, a DivisioniofLoews Theatres, Research Department, Greensbono,. NC 27420 Reeent technological advances have given..chemistsgreater access to inexpensive an& powerf'~ulicamput'ers. One applicat'ionunder investigation util'izing,computers in our lab'oratoriesinvolves a search for correlation~between~stlructural features of a flavorant molecule and iRs per~ceived'flavor. Structure-activity rel'ationships involving.steric, Hydrophodic,, electronic, and other molecular parameters have been widelyy studied,, particularlyin the prediction-nd synthetic design of new pharmaceuticals. While the mechanism of human olfactory response is not known, most current.theories invoke thepr:incipie of a molecule fitting into specificc reeeptorregions and initiating a specific olfaetoryy response. Our work has involved the development of methods by which the surface regions of molecules can bemathematically expresse&, compared, and visualiaed~in aa manner allowing forr predictions of receptor sitedimensions.. Sever,ali molecules.sharing similarcedarwood flavor properties were examined by computer for.similaritiesin shape. it was demonstrated that cer,tain regions were similar in contour.. A proposed! organoleptic templa.tewas further tested by synthesis.Evaiuation of the utilityy of this approach to flavor chemisbryy is currently in progress. 29
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FRIDAY MORNING -57- ~ THE CONCEPTS. OF FLAVOR IN THE CIGARETTE INDUSTRY. F. Abdallah_, 10:35' A. C... Monk.6' Company.,. Inc., Farmville, NC 27828 Tobacco is defined in terms of seed,plant, cured leaf and varibusproducts- with more emphasis -smoking,p,roductsin general andicigarettes in partic.ular. The role of genetic factors.,, environment, cultura]/management practic.es,, harves.ting/curing.,, and aging/fermentation on the.chemical and physical make-up of the leaf will be discussed. Besides chemis.t'ry.ger se_,, other f~actors.such asf'~ililing.p,ower,.moisture holdi'ngcapacityy and burning: qualities contr.ibuteto the basic characteristics of the cured leaf. Siheethe actual prod'uct consumed i~s.ttie smoke, the complexx parameters involved in the.smoke formationwillibecoveredi. This will includebot'.h tobacco and',non-4obaeco parameters (additives, paper permeability, fillterefficiency)r which alter the.physieal and chemical eomposition.ofn thesmoke.. The concept of thee cigarettemode.l, in terms of physical and chemicali characteristics of the mainstream smoke.e will be reviewed. The smoke-smoker relationship, wi~ll be discussed in terms of'~ both flavor detection and the physiological/psychological effect of nicotine and related alkaloids.. The importantt roleof'~ flavor in the evaluation of; smoking,produets will be discussed andlemphasized in the development of low "tar" and nicotine cigarettes with acceptable smokingqualities. Thee chemicali constituents of the mainstream.smoke - espeeiallyy thetotalparticul'ate matter TiPM - and these.Pective.contrib'ution of these constiltuents too smoke fPavor will be reviewed.. Today,"s achievements in thef'ields of smoke analysis and fliavor precursors are.a.ime,lnot'onlytowards understanding smoke formation but also towardsmodifying and.improving.the.flavord qualities of thee smoke. 10:55 CONFERENCE ADJOURNMENT 30 Q
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AUTHOR INDEX (BY ABSTRACT NUMBER') Abd'allah, F. 57 Abel; S. G. 19 Adams, J. D. 49 Arcila, J. 10 Arrendale, R. F. 23, 32, 43, 44 Baker, J. L. 32 Benson, A. L. 55 Blume, D. E. 9 Bohlander, P. J. 16 Borgerding, M. F. 38 Brummer, U. 28 Brunnemann, K. D. 33 Bush, L. P. 4 Carmella, S. 50 Cartwright, W. 40 Castonguay, A. 49 Chamberlain, W. J. 32' Chaplin, J. F. 42, 43, 44, 45 Chortyk, O. T. 11,, 23, 43, 44, 45 Collins, G. B. 13 Cornell, A. 40 Counts„ M. E. 53 Crutchfield, J. 41 Davis, D. L. 13, 30 Dirks„ V. A. 12 Dube, M. F. 5 Dwyer, R. W. 19 Einolf, W. N. 36 Eivazi, F. 41 Enzell, C. R. 29 'Presenting author in italics 31 Ferguson, R. N. 36 Flaherty, K. A. 51. Fleming„ M. L. 19 Gord'on, B. M. 25, 37, 38 Greene, G. H. 25 Griest, W. H. 35 Gwyn n, G. R. 11, 23, 43, 44, 45 Hamilton, J. 31 Hecht, S. S. 50 Heckman, R, A. 5 Heeman, V. 28 Higgins, C. E. 35 Hoffrnann„ D. 33, 49, 50 Hyde, R'. A. 18 lhrig, A. M. 48' Jackson, M. D. 44, 45, 46 Johnson, A. W. 43, 44, 45 Johnson, V1/. H. 10 Jbnes, S. T. 2 Katz, T. 36 Keith„ C. H. 22 Kenefick, T. 55 Kinser, R. D. 36 Kung, S. D. 6 Lam, J. J. 9 LaRoy,,B: C. 53 Larson, T. M. 48 Lee„ S. J. 49 Lee, T.-H. 52 I
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Leete, F. 24 Legg, P. D. 13 Leggett, E. 31. Litzinger, E. F. 17 Long, R. C. 3 Lynm, D. 5 Marmor, R. & 56 Mason, P. F. 44 Massie,, I. E. 1 Mathis, D. E. 15, 21 McCarthy, R. 55 McDuffie, K. L. 11, 44„45, 46 McMurtrie, A. 17 McRae, D. D:. 20 Mohapatra, S. C. 1i0 Moss, B. L. 48 Nishida, T. 29 Nishikata, Y. 14 Nordfors, K. 29 Norman, V. 48 Olender, V. 40 Olerich, G. 35 Pandeya, R. S. 12 Park, J.-C. 52 Perfetti, P. F. 54 Perfetti, T. A. 26 Pitner, T. P. 36 Poushinsky, G. 12 Rhodes, P: R. 34 Ri'vers, J. M. 5, 39 Samejima, T. 14. Saunders, J. A. 34 Schepartz, A. J!. 42 Severson, R. F. 111, 23, 43; 44, 45, 46 Sheen, S. J. 8 Sims, J!. 311 Sims, J. L. 41 Slaven, R. W. 27 Smith, L. B. 11, 43 Snook, M. E. 11 Song, B. H. 30 Spears, A. W. 2 Squire,, K. R, 47 Sutherland„ P. 55 Tso, T. C., 6 Vinchkoski, N. 49 Wahlberg, I. 29 Wallin, I. 29 Waymack, B. E. 47 Weatherly, C. H. 21 Weeks, W., W. 7, 30 Weybrew, J. A. 3, 7 Wilson, R. L. 42 Woodlief, 1N. G. 9 Wu, D. T. 17 Yoo, K.-K. 52 Zilkey, B. F. 12 32 l
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35th Tobacco Chemists' Research Conference October 7-9, 1981 1Ninston,Salem, North Carolina Conference Chairman: R. E. Noft/e PROGRAM EDITORIAL COMMITTEE D. L. Davis, Chairman E. F. Litzinger C: F. Mattina POLICY COMMITTEE W. A. Selke, Chairman R. W. Jenkins & T. Chortyk E. A. 1Nernsman A. Rodgrrnan LOCAL ARRANGEMENTS COMMITTEE B. J. Annese H. W. Baird D. B. Fishel P. G. Goforth R. Hutchens J. Lawson L. S. Lewis E. A. Li A. R. Mitchem J. A. Nowelli A. Piehll A R'odgman D. E. Townsend FUTURE MEETINGS Raleigh, North Carolina, October 25-27, 1982 Host: North Carolina:State UniversiYy
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