·12;11·:~·-. i r g Aw~341 ·: 5:i: cG`fgtit~ ITE~P~H$t~Ea~k: F:-· nc, .? ~CS ::t: ·· ,·- ·I 1WI I'YB~~~ht~A'~i;;;~h~B%")S~P~h~~ll~';;YII~·~n)~-~~ili;·'vi ... j i, *: laTPli,RESIARUI IABOApaOAY· tm, ,. 64 i Acetate·, Inhlblticn or Radllll Inltfat~d~ ·lc: i 1 i Projectl 7705 R1 He, 138 , :: 741':"''I~" :u.I '~: 71··!1 :~~·4~ ·· ·:~ .r ~·: "'I·~B i~lp*~'~;U;U~PB~.\~~~b?86~r~!i~(uc9Ri~5~c?~HS~I if: ·~ ·· I I· :th~il ) i··.~: - ::-: ~v:D;' 4·i~ b': $·! c, ,i ·rr· ;Y*Y;I ~T~F~ i4:~".~i~im~3WP~ge~S~t~a·"f~~I;=::'; ~i~%.~B~~18~$L~s~6~wrPB"W~b~"Y~3~l~jl:~~~Wj~;~;~~ :I : N m~lr·'~I~?~SL~X;eb~O~t~al4~wa~u~wcx;~c~;i~~s~iit~~~~':r O CCI \O O 0\ BATC6 document for Legal Services : Health Canada 20 May 1999 Ar ~tYL ·tC~c·z~t~4~ ,~ ~e (fL~bl~ BATCo document for legal Services : Health Canada 20 May 1999 P, 10, B, ~BRII1Y B,A,T, CO, SODaMMndl III~~~X~~~ BPd ~!~ id l~lL7~hj i Illull ?~~ 1~8 Dots rs~i~a~: I '~ ril iv Bspdsr lats ILitilla Dlt~ IP rOD 1~T TO non~ TIIrS alonr To uo~n ~~s, P~~ IISEBT I~S~CT !~B1LS bP~B 'IIB~~' ~t T~ SE=T CIBC~~TZOB BLOCI, ~S EISU~TG ~ICI ~CQLL ~B~ TT IS QEDBD """""'7""'-"'i.'-'~"'.~.i~~r-~ ~lle· b.ii ~e 7i 1~( ~J 9c~ d3-·rl~ (w LL~I z kai a BATCo document for Legal Senlces : Health Canada 20 May 1999 _ __ RESE~RO LABORAtORY REPORl MO. 138 RLB ~nPERIAI tlleCO PRODUaS LInITEO LSEIM lAOORATORY RITE~L IIHIBIT1DI OF IleAl laI~I~D POUnERfS~tIOI OF Ylrll ICETmE BI TOAttO SnOxE ~O 5oME Polrnctre HroRocAaoa PROlFCl T 7705 ~AH~: 01, A,A, Mr50ET RS, S, S885l8llER MR5, J, JM~OI OR, H,H, B~LfMORIA ISSUED BY: DR, T,d, SMIM DATE ISSUED O~~ER 6, 1972 OfSTRIB~la: i, library 14, Or, S,J. Gncn 2, Mr, R,M, Gfbb 15, Dr, S,J, Cncn j, Mr, RS, Vlh 16. 0~ 5,J. Gnen 4, Or, T,A. Smit~ 17, Or, S,J, Onen 5, Mr,S.M, Candlish 19, Dr. 05, Fc~~n I 6, Dr, M,A, nisbet 19, Dr, D.G, fclton J 7. Mr, R,1, Rice 20, Or, f, Hlslam 8, Mn, J, d~nson 21, Dr, I,Y. MugheS 9, Ma, 5. Schlncllet 22, Dr. R,A, 54of9rd 10, D~ LH. Hnm~l 2J, O~I~, Sarford 11. Mr. E.R. F~lerldcn 24. Manager; RLD Austr41il 12, Mr, M,P. Sc~l~ak 25, Manager; R L O bustr~l~ 13, Or,5,3.0re9n Idorato~ ~tcb6ats 13, 312 BATCo document for legal Services : Health Canada 20 May 1999 SLNhiARY A b~ide ranoe of organic ccn~pounds, which a~ caDable of intet· teptina iree radical intermdiates fan~d during radical initiated vinyl acetate p0lymcriration, have a Mined impact on the rate of polynriration of vinyl acetate. The efficiency with which a conoaund retards aolvmeriration can be estimated by Rasurina the tiw it taker for a mixtu~P of rnonb~oer and ~eaavlleroxide, ianersed in a Hater·bath at MOC, to reach a IPootaneovr boil and ermnarinP it with the tint! readnd far a iimilar mixture with added starter to reach boiling paint, We have tested the efficiency of smokes from various sources as inhiiitnrr as well as a ranoe of polycyclic hydmcarbons, Smoke vaoour is strongly inhibitory and the activity has been attributed to eonjuoateb diene~ thiefly iropnne. Smoke condensates from different tobacco typer can 1e Ciffenntiated by their relative efficiencies in retarding poly- meriration. Flue cured tobacco smoke has the highest activity: smoke fTOn reconstituted tobacco has the Icwert; a~r·cund, stem and pe~que tobacco have internediate values, The effect of the smoking vehicle war mearureC oy tertinq Cigar and oiw smoke cPndensates, The ionr has a value similar tn sit-cured tobacco but the latter has 1 verr 101 activitP. Ofthe nalycyelics which we scanned, cateino0enic subrtrmces ~enerally ih~h~ed hipher activities than nwl·catcinoqenr. sane weak carcinogens such as anthraeene Here, however, une;coectedly active inhibitors , A Sydrocamon enriched fraction of smoke did not /epear any an active than a crude condensate, BATCo document for Legal Services : Health Canada 20 May 1999 Be Eclanirna by uilich sndre inhibits po~y"e'i"tion are diicursed. Uhile all three possible n~chanir~s; rulrti:utjnn reactions, stat ttansfrr and radical coupling pra~ably oreu~, i: is portulabd that atom transfer is the dominant one. P BATCo document for Legal Services : Health Canada 20 May 1999 ·_ I~SPi~R~ fine ~n,te Reihe a,aanircoer Substanren, welche irl:anee sind in einer vcn Paditalen eincleieite:e Vinvlacetat-Pol~~ririeruno ea:staodene !reie int~rrdiar Radilaie auiluhalten, haben eineo auff~lienden iinfiurr auf die Poiyn~ri sati anr~roinindi gkeit van Vinyiacetat. Oje Leirtunorfihigkei t mit welcher eine Sujrtan2 die ~olyeririerung ven89ert, ist leh~2bar aJrch ~errunoen der 2ectintervdlles welches e~arderlich ist un ein IDcnt3ner Siedel iu*r ein Secnqe von HonOme~ und BenzovlDemx!d, eMilmt in einel Uasrerbal van 70'1, 2u erreichen und durch Verq~eilhen derrelbel ~t ihnlichen Gewncen zu velcheo 5uSstanzen lit verz~elder WirkunP hinluoeeejen rurden. Wir ori~ften die Leist~masi~hipkeit des Pa,(her ,eri~nieitcen U?rP,Eo~i, rmie eine Reihe lyklircher Kahienrarreritoiie Ravdl·[ia~ofohdse ist ein starker Inhibitor und seine 4ktivi:it is: den koajuoierten Oienen, haupts%ehlich ~e~ IlhDRnl I uTUS dr re i bcn . ?aidlkonCosate van verrcniedenen iajaktypen sind unterrcheid- 3ar ds,cr iil~ reldtiv untenlhiedlilhe Leirtunosfihi gkeitsven~enmq i~r ?claP~irienng. RauCb van fluecured Tabak hat die hlc~lte Rltivitlt: iru[" von re~lL~rtiaiertem iabak die niedtigrte, Imd der van air·cund, jtrunk· ~na Perig~i :abaken hat intermediit Aictivitatswerte. Sic Ui~lno der Rdue!lvehjkels wurde uenfiit:eilt Pro~en van 2iPrnl· unn Pfeifenraucht~ndenraten ge~tren. Oas Er',IOenannte hat Jinen a"nlichen Yer; wie a~r·cured Tablk, das letl:ere reda~ i~lt eioe ;ehr nic~rioe ~tti~~tit. BATCo document for Legal Services : Health Canada 20 May 1999 _ __ Unter den ron uns ~prJRLn lyUlschen K~~nwarlcntof~n nl~a unaose er~as ~*nn ~r ~~R L~d~t BI $dt~namae. M~r kidD enc~~~ ~loQ, kbnl· LI, wann unemartet aCtive Inhlbit0Rn. ~ne ~ t K~]enHaSle~tOff n~~iche~e Rau~f nt~M I~ien ~d~ ~dvcr ~I ~n Rlucnkand~sal zu I~n. ht ne~l~l·l ~t rclchel Ralcb Me rd~~n~lr·P ~M~e~ Murde disku!ie~. B ruth ~ert~ic~, d151 der #odrllrpo~ hr daddenl~ C~llr*I ~n hi dni kJbr~Laddnl~ ~o~bPh" Rt~db - Su~r~tJe~n~ bt~transpo~ ~d Ra~k~kupplun4ist, B~Co document for legal Senices : Hea~h Canada 20 Y~ ~999 1~5VY ii, ~I:e ~~~x de eornorC~ arpa~~ues caoahles d'intereeotcr de~ irtermeaiaires ~bcaux librcl fonis lnrl de id o~~d~~~on ra~caiemln: initiee de I'acCutr de vvnile, ont on erfet ~rovC lur Ic tiux de o~~· ~~~oa de I'a~~ta~ de vvoile, l'effie~it( d'un eaDalC I retarder la sl~C~wdon *d Ftn lal~e a m~llnrt L h31 nPdl *r cc IP~ tidn d'un mon~~rr c: de pcr0xyde de benlovle, ~anoCe dens un hair ~ !~"C, 3oUT dtt~~re Son oaiot d'C3ullition Ipbntank, au temrl reouii oar un m~anoe 310~irl Id~n0nnj dl~n retardan:, pour atteindre Inn roint d'C~~- lition, Nour avanr cxami~ l'efficadlC ~e fumkel de lourcrs veriies i I iteurl, id einsi quc nr~in~ hydr~eatbones p~!cv8iouls or tant ~u' nh b aQur de f·k at fDrtement CMM~ke: we rMdtC I ite ~d~bil i der dihres co~uou~ orincloaiemert I'i~anrln~ Ocs eondensCf de fun~ orovenant ~ diffe'rentes sortes de tabac neuvent ftre ~fflnn~~ selan Icur cfRa~tC reraertive b retarder la ool~~sn~on, la fumle de tabac jeune a ia ~u~ gnnde ae~~t~: id f~C dP tl~ll reConStifUe 1 Ia IijUs harie a~~~tl; les tabacs '~t·eure~', ~~oue, et de dne, ont der acthi:Cs lovc~ner. l'e'fet de ~a fone du nroGuit fwne a etC mesurb e~ cx~C~ment~nt ~ur des condenrls de f~~e be eioates et de piaes, re p~emier donne deT n- s~tatr lee~a~er av :abac "~r·cure~'.~ir ie der~cr DOIS~de une dctlvite' trPs aarse, Plni let nnlyevelioues Ixa~nb, les cormesbs cardnnolner oCro~ trent une ~ur onnde IIM~tC ~~ Ic~ e~D~Cr no~careinan~ncs. 4~]p~~l carc:nookoes eonsidCrCI eemoe dtant f~~~l, t~ nuc Ilanthrac~ne, se rant toutefais dbontrCI came ~tant de ouissants inCihiteur~, BATCo document for legal Services : Health Canada 20 May 1999 Une fractinn de f~hlre entiehie d'hdroearbOnes re r'est naS montree nlus active oue le eondensC brut, Les dchanismer oar lerguelr la furde inhibite la nolvn~risation Icot diselter, Alorr oue ler tmir ~dchanirmer rarribl!s: ler re'a~tianr de substitution, le tranrfert d'atane, et le counlaPe de tadicaur, ant pmba- blement tots lieu, ii ert noJtule/ Pue le transfert d'atome est le mhchanif· me dominant, BATCo document for Legal Services : Health Canada 20 May 1999 INTR~?1I[T!ON Tne~ is continued intea: in finding chemical or biacnemic31 Dredic:oa of the biological activity ef tobacco smoke, Che appmaeh to the Droblen nas been to select enpnes which are responsive to snake and to bare a predictive test on enzyme inhibition ii), it is oenenlly accented that inhibition, in most cases, is due to reacticn of sloke with lulfhydril group~. Evidence for ti~il has come f~dm studier of inhibition of aehydrogenaser by snake (21 and fron the fact that the presence of added cvsteine can decrease the inhibition (3), Our own approach to the problem has been to develdo asaYS for Eoeciiic Dronertier of smoke; properties which He feel an of biolodical inportance, and an attributable to a class of canpounds rather than one roerific Caa9ound. An example is the redoa property of smoke, which can be n~dluted b? the ability of smoke to reduce dyes (4) or olidire nscorbate (5). Our attention tar drawn to ndhx charac!e~lstics durin!: sm studies on the effect of smoke cn bioiopical electron transport systenb, It war, found that some of the disruption of the system could be ascribed to the reducticri by smoke of electrca~ accepters such as cytoehm* c. A second oroperty on which we have focussed is photorensitired o;tidaticn of aaeovs ~replratias of smoke (61, Condensates contain both seori:isers and the oxidisable substrate. RDpreciable oxidation rater lan be achieved by irradiation with light at wavelenpthr areatet than 301 n~. RdCition of arco7bic acid as an auidisable substrate considerably enhances the rate of oxygen uotake. This latter reaction iupgerti the oo~rihility of a test for the bialo~idl activity Of Smoke bared on the photodynanic action of aqueous O~paratiani of condensates. Ph0tedynamiC activity has P BATCo document for Legal Services : Health Canada 20 May 1999 __ ·2· been defined as photosenritisation by substances not no~~~ present in 1 aiolc4ioal 5.vstem ~]· net of the investigations in this area have involved Kdftldll dyes a~ dcn~n~a, PElaxide RnlHa is believed ta be the reactive intenediate involved, Gcntinuiro our search for li~le predictive tests, we hew recentlv ~jkd ~r effect of smoke I radical initiated p~~e~u~a of vinyl lcetatc. It has been rh~n that certain classes of empnu~ 1~ oo~~ af nPrP3 benloyl pndh initiated Idy~ldlo Qi ed in ~ct i nu~er of anticancer d~ps have been demonstrated to have strong lldical inhibitory properties, It coulC be ewpected that sore ai tne reactive qedu ~ Iloh nnuid be PPIL If n~dal with free radicals thus retlrdinq vinyl acetate pol~rlsati~. Udnp the standard procedures described by teor4ieff ~il ~ have Imcniced the cfhln of lab and 1 a~r of nnlycvciic Imdla on this n~~m and the results 14 n~o~ed in thjs paocr. 3aterillr Ind Yethnds Yinyl acetate, free of stabiliserr, was ~tdnrd fr~ G~f M1 (Canada) ltd, Benroyl p~~de Mar crjrtalliled ba detofon lrio' t" T~ nolvaroMtic hydrocarbons Hen tested far purity by nelting aoint and thin.layer dmll~qrll!; Mhere necerrlry, la Here cas9nlwd to conrtan: oeiting point, OaPan was scruoulauslY cleaned by a se(uence ~ ~ahhg in aerhanolic pta~o k!dn~db cnrwllic Kid, Mnlnd Mater ad finally I~an. me tuber Mere dried and stored in 1 dust free ~arm·~tr Oven, BATCo document for legal Services : Heath Canada 20 May 1949 __ ·3· Cioaretter and ~iparr were smoked to the standard naia of one minute to butt lenothi of UI 4nd 50 nan !5 mi puff of 2 second duration Oer RIDee:iYelY· Pills vrR ild~~d on a i3etially lonrtncted machine drawing one ~5 nl puff of 2 second duration at 10 second intenaii. Plr?iculate matter war collected on weighed Canaridoe filter pads for tests m ~n· irac!ionated smoke. HydmEamon enricned fractions were prepared By the ne:h0d of Giim~r (1Pi, ior this iradiooation the cendmlate from 50 - 15 cigarettei or 30 - ~ cilarr rlr collected by eltrtmiai~ DrPliFitlTiCn· Yutioni of inhi~itorr were prepared in vinyl acetate, When rse sole Has :rapDeo on a iadridg~ filter,i~e entire lad war acera?Pd in the ranbet and filter fragnents were removed by filtraticn, if IRDte frictions or i·vdrata~anr nPre being tested the la'srial war dirralvld dirrtl? in vinil aarate. The concentrated solutions were all made up to gtai~ 5 4 of inhibitor ~er mi, 'he tests were tarried out by placing 10.00 mi vinyl acetate and !3 mg of beniovioomride in 20 x 150 an Ilair tubes. Ouolitates win Ireoann ~Y adding 0.0t, 1.0lt 0.03, 0.05 and 0.01 mi of inhibitor to mnaaPt immersed in a constant tenperature Detoxid~ ni ytu~. The ten tubes were oath at 70 t n.j'l and me tie required to reach a roantaneoui boil was recorded. The presence of a control with each test set war designed to counter the effect of iluc:uations in bdth temperature, The tests on vao0llr phase were achieved by crauring the rmdwt from the ri~~rettl through a (ind~e filter d:re2ly into a prewashed tcr:tube lontainins R.4 mi of vinyl a~ctaa, cooled in an itc·bath. (iee fioure i). BATCo document for Legal Services : Health Canada 20 May 1999 -41 ~igurr 1. VapPu, phase trapping ryrtem, ~,, cigarette C-----t Il--cambridge filter to smoking machine I3l~lvinyl acetate BATCo document for Legal Serrices : Health Canada 20 Hay 1999 .5. be to four puffs, excludina in all eases the Ilohting puff, were bubbled into the monamer. The tubes were then removed from the apparatus, 2 mi of vinyl acetate containing 20 mg of bentoylPeroxide wece added and the mixtures Here inn~rred in the water bath, the tire required to reach a spontaneous bail was recorded, A control was derived by bubbling the aporopriate nuRaer of puffs from a unlit cigarette through the trap, Least squares lines were fitted to the data and from the slopes o' the lines, tne inhibition factors (delay in spontaneous boiling) exoreried as minutes per ppm, Here derived far solid or liquid inhibitors, In our tables, to facilitate visual comparison we have expressed the results as Pin ppm .1 x 1o3. 1, the case of vaoour phase the results a~e shown grapnially as minuter per mi of bubbled vapour, Results and discussion Our initial studies were directed toward smoke vapour phase which we found was an effective inhibitbr of viriyl polymerisrtion, As can be seen from the qraoh in figure 2, se puff of sake is capable of delaying the soontaneous boil b? 4 minutes. The extent of inhibition sho4ed a linear relationship to the number of puffs over the range which we examined. Information as to the type of canpbunds in smoke vapour phase sast likely to contribute to this inhibitory effect Has provided by Georgiefi's study of the inhibitory properties of a ran(le of unsaturated hydrocarbons (9), He observed that a nu~er of canjugated diener and triene~ were oarticularly active, The vapour phase of smoke from flue·evnd tobacco contains the followinp dienes. BATCo document for Legal Services : Health Canada 20 May 1999 -6- Figure 2. Effect of smoke vapour phase and isoprene on the polvmeritation of vinyl acetate. seconds 1000 sao 100 700 600 loa 400 300 200 a smoke vapour phase D isoprene 100 0~- 20 1 40 60 1 80 1001 120 140 mi 2 3 4 puffs rV BATCo document for Legal Services: Health Canada 20 May 1999 -'I- CoSound Cooeentrati on ( 40 mi puf fi 1,3·butadiene 4.3 e9 iroprene Pf,O u9 I,]·tranr·pentadiene 1,2 sP l,]·cis·oentadiene 1.9 vg The major c0npcnent, isoprene, was tested far its inhibitory activi:y over a ranpe of concentrations similar to those Encountered in r~oie vapour (see fisure 2). It was found to have an inhibition factor of 782 min opm'l n 1(13 and calculations showed that Isoprene alone could account for 92: of the inhibitory activity of the vapour phase. If one ass Ud Ijmi )aT inhibition factors far the other dienes ansent, 95: of the retardation of polyretiration by vapour phase can be attributed to its dlene cwltent ~ Condensate, af unfraetianated smoke were tested by MceratinS the Can~ridge iiiter oads, on uhieh the smoke had been trapped, in vinyl acetate and filtering the solutions to remove fragents of fibreglasr. Estimates of the drl wein~ts of material on the C~ridqe filteruere calculated from the predicted water content of smoke from the various sources. The smoke from 1 range of different tobacco types and smoking vehicles an compared for their abilitg to inhibit palymerisation; the results are shown in table I. BATCo document for Legal Senrices : Health Canada 20 May 1999 -8- INHIBITION OF POLYMERlSAflON OF YINYI ACETATE RY OI FFERENfmBACC(ISM~E tMI~E~T;AT~ TABLE I r I. I Tobacco Fonn Inhibition Factor mi '1 x i~3 ppm Flue Cured Gut 115 Flue Cured Granulated 114 Rir Curec Granulated 87 Stern Cut 98 Stem Granulated 95 Peripue Granulated 82 Recortituted Cut ~1 Re eer ti tuted Granulated 66 Pipe I Clear 90 / vlridtion of 2.1]2, -- BATCo document for Legal Services : Health Canada 20 May 1999 ·9· Comparisons an made be&cen the same tobacco types in cut and granulated :an so that s~all samples of to3accn, of which inruff'cienf material is available for conventional cigarette manufacture, can be included in the Lest series, The granulation technique consists of grinding the tobacco and collecting the fracticn which pastes through a n~et 20 and can be retained on a nla~er 35 mesh sieve (iyler Screen Scale), The ground material is then oound into standard cigarette tuber rith a cellulose acerate filter, The yields of particulate matter from cigarettes made in this fashion have Seen Ih(wn to be comoarable to yields iKwn conventiodi cigarettes, Tbe reslllts indicate that there is no sianiticant difference in polygrira~ion inhibition factors between conwnra?ei from the rare tobacco in cut and granulated farm, The results also shoved that flue·cured tobacco has the highest activi;·i with a value of 115 units uh'le pipe smoking produces the Imest, !3 un.:s, intenediate values were found for air-cund, Item, reconstituted tobacco sheet and aeriqw tobacco, Cnnsideralle significance has been attached to the presence of rnlyamadtic lydrltrrbcns in smoke, it is cmridend the: the Dolyamaatics ate responsible for the greater part of the tumorigenic activity of smoke as measured by mnure·5tin painting (il). Smoke was processed by the mithod ~lcribed by Crinmrr (12) to yield a fraction enriched in palyaroPatic hydrcl· ea~ons, 1 nurbet of these fractions were preaarPd add tested far their inhibition factor:, The results are rh(kin in fable II, BATCo document for Legal Services : Health Canada 20 May 1999 110- INHIBITION Of POLYHERISATIONBY POLYAROIIATIC HYO00cARBC6l NEO SMOKI I nnL i 1VIIJ VU i n~ - - TABLE II Inhibition factors Source of Ftactio? nin oorl x103opt" Flue-Cured Flw·Cured ~alvene Extracted flue-cured Acetwle Extracted Clear 1 112 i~---------- BATCo document for Legal Services : Health Canada 20 May 1999 -11- It is noticeable from the table that the enriched fractions do not have greater activity per ppm than whale smoke. Analysis of variance fndicated:hat no significant differences existed between the fractions derived from various tobacco typer. The low activity encountered here suggests that there Is present, a large amoPt of inert material, such as aliphatic hydmca~wlr, which does not inhibit ~alpriration. We plan to purify this fraction further In the hope of detecting qullltative differences between Smoke from different sources, To provide a baris for comparison we have investigated the inhibition factors of sae aromatic hydrocarbons including both carcinogenic and non- carcinogenic conpounds. Table iii sunmarises the results, The Careinogenidtv ratings given to the ca~ounds tend to be subjective since the hydrocarbons have not usually been tested for tunoripenic activity under the same elpen'm;ntll conditions, Na direct correlation can he drawn between the ability of the hydrocarbon to inhibit vinyl polynerisaticn and carcinagenicity, I: can be observed, hmver, that the active carcinogens have high inhibition factors ranging frtn 272 units for 20·methylcholanthnne to &69 units for 9,10·dimethyl-l ,t·benz~nthracene~ Weak ·carcinogen~ tend to have lower activities~ crmpo~ds, for example anthracene, do 1 demonstrate a high ability to read with radicals formed during palyreriraticn, but fail to behave as active carcinogens, n·.'2-~~ Cp~':ah "E~ Y~pl 12 ,g6& 4 'c~L p a~B-~ o hEa-i ~· r BATCo document for Legal Services : Health Canada 20 May 1999 -12- dlalLdOn r ·~16 11101 ILln 16 WOLCII lalrlAltv rvl~ntrl,nllr' n 1111I~1 TWLE fII tlrdnopcd d~ Inhibitian Factor ~'1 ,103 min ppm 12 Phenanth~ne Haphthalene t 53 4,5.Benloo~nne Chrjsene 141 PvRne 1 ,2 ,5 ,6 · M bmz an th r aeene Bb~~dB·Ln· D1 352 I,t-lenzmthracene g.10·0i~thylbnth raccne 0 396 Flua~an~ene m a55 3,~·0cn20pynne Antnraccne O 697 3·~thy lanthracene g ~~.M mthyl ~ ~dFnlan~tacme H' ~I Ic~ve car~noF"S~'Weak carcino4ens 'Source: Ln~ af ~I·I ~d Lr Lr iii fpt hd·lic ~~,~~ plm~M nn I L~1~~ Z 1ZC~ BATCP document lor Isgd Services : Hedth Csnldl 10 kley 1009 .13. Rn explanation of why sore aromatics are so mud! more prone to addition of free radicals has been given by Coulson (13) who has calculated I ocalisati on energies for a nutter of arornati c hydrocarbons, The "pteparedness " of the hydr3ea~bon to react with an approaching radical to fow a new c · c acnd depends on the ease with which 1 electrw can be lacalised in the hydra- carjan, Sware (14) has confirned this Horl; cxperientally by r~asuring the rates of addition of methyl radicals to araatje eompowlds, It was demonstrated that the logarithms of ethyl affinities ate linearly related to locatisation energies and also,that logarithm of the relative rate constants of addition of any radical R' to a series of aromatic hydrocarbons bears a linear relationship to locdliration energies, The relative abilities of the aromatic hydrocarbons to inhibit vinyl Doly~nerisation closely parallel their relative reactivities tward methyl radicals as determined by Swarc. As a basis for interpreting our results with smore and arocnatic hydra· car3onr, an explanation must be given of the type of reaction Imder study. The reaction n~eci~anism in the ~Rsena of an added retarder has been given by Kice (15), Mechanism Initiation 1+2RI~P' Propagati on P'~ M ~ P' Trans fer or Addf ti on P'~ X ~ 1' CooolFeri s ati an Z't M ~ P' Termination 2' ~ P' ~ polymer 1' ~ 2' -r Inactive oroduct P' ~ P' + Polvat I,initiator, M:ranoarl X ~ ~tdrder, P ~ oolysr, P': Polyar radical, h~ 1': radical fonned by the reaction of P'nith retarder X~ Chi BATCo document for Legal Services : Health Canada 20 May 1999 -14- Three tyoes of reaction can he involved in the retardation nrocen. i, P~di ti on o r Subs ti tution Sore substances react with radicals to nroduce mother radical which nay be too r:able to enter the kinetic chain or is rebcti w alanp a diffennt pa:hray. Examolel of this type of reaction have been quoted by Sware (151 who StuCied the ~activity Of aromatic hydrocarbons tcmard ethyl radicals generated from acety~peroxide, g' ~ C6HS * RCsH6' Substitution 1C6Hg' ' R~i RCsH5( RH Addition a H " n R H i, Atom Transfer The most commonly encountered transfer is of hydmpen Which takes Ilace from comooundl such as phenols or aminer and other naterials with active hydrogen, S' ~ H-D · SH ~ 0' 0' + new reaction H~ represents a hydrogen donor. 3, inhibition av Radicals .4dded f~e radical species car intercept the radicals fan~d Ss~ BA~Co document for Legal Services: Health Canada 20 May 1999 ·15· during the oolyneriratian oroceii and produce inactive products, euamole of this inhibition tvoe was dermnstrated by Ilartlett (17) who showed that diohenllpitrllhydraryl , a stable free radical, was a very effective inhibitor of radical initiated vinyl polymeriSation. fhe reaction can be described by the equation: R' ~X' +RX Retardation will occur provided that the species It' cannot react with R to produce R', The infoonation about the three mechaniss of inhibition enables us to predict the classes of smoke comDoncnts which say be resnonsible for ntardino vinyl acetate oolyn~risation. This is shown in Table IV, BATCo document for legal Services : Health Canada 20 May 1999 _· '16' M~CHPYY~SMS OF IHHIDITION OF RADICAL ·---~M~E~jiijR';iT1~icD~N~~;E INI....._) POLYP : ~31·_ - tABLE IV lnhiliti on mechanism Reactive roecies Exarrgles in smoke Addi ti on Irubsti tuti on dwble bonds unsaturated hydrocarbons aroatie nuclei arwMtic and polyaron~afe compounds , j,,,;n,, active hydrogen phenols an~ines nducino spents Inhibi ti no 31, radi cals radi cals resonance stabiliseC smoke radicals I ------------- BATCo document for legal Services : Health Canada 20 May 1999 ·17· More specific infanna:iofi about inhibitory COmPOlpldS in Sm3ke can be derived from the systematic studies Sy Geargieff (18) an substances which retard benzoylperoxide initialed vinyl acetate polymtisatian. A selection of his findings is given in Table Y, As the table indicater, most of the active inhibitors tested by Georaieif have been detected in smoke, Chlombutadiene me hexatriene have not been detected, Hlmever, other conjvgated unsaturated compounds an known to be l,nsent 110) and inhijitiln due to this class of ecn~0ounds could be exwcted, The free radical diphenylPicrlihydrazyl is sat known to exist in smoke but other resonance stabilired radical sneciel do occur and have been demonstrated colorinetricaily (20) and by ESR spectra (21). They are sufficiently stable to exist far several days in smoke condensates. Our extension of Gear~ieff'r test to scan a range of oolyaromatic hydrocaroans indicates that while sorre polycnndensed eanpounds, particularly earcinoPens, have high inhibition factors, me concentration of there materials in smoke is too low to make a major contribution to the observed activity. Sone exgerinents in this laboratory and elsewhen an the reducing oropertier of ~a~beeo smoke (4), coupled with the fact that in the food industry the Dtesenative action of smoke an cat is partially attributed to the antior.idant effect of phenols, suggested that hydr0wn donors sight play dr: important role in sae inhibition of vinyl polvn~eri~atian, To test this hJ;iotnesis, He cnmeared Be results obtained for dye redultitn by diffe4nt smoke condensates with the inhibition factors of the sane smokes, See Table VI~ BATCo document for Legal Services : Health Canada 20 May 1999 -18· latBnIOI o~ POLIEdS~IOI BY CMPOAOS~L~ TAB1E Y toqound I nhihi ti or Factor Pnsen: inSrnoke (19) ,1 min ppm x Unsaturated iiydrpcarbons ~exene-) 2 I~·Hewadcne 9 4·Ch lo~·1,J·butadlene 521 trans·1,3,5·hexatricna 1570 Phenols Hydr~~~one 1010 I·B~~rtylabcb~ 920 1~16.i~hyd~wy~nzene M10 ~~inoncs rl·~da·e lp t AmineS Oionenylamine 770 Id~ln~yla~~e 700 [arbonyl compounds Acetlldchyde C~ton~Cc~ydl 62 Acctle Sulphur compounds Thi alene Bcntene thial 110 Stable Free Radicals n 2~·~phcnyl~·~cy~ hydrarll 1170 N BATCo document lor legal Services : Health Canada 20 May 1999 ·19· LATIMI BE~WEEII OYE REDUCING RO INH[SITIOM OF CORRE OF TOBACCO SMOKE CO#OENSAtES P9!YMERISAtlOll ICIIYITIES TABLE VI r~ Flue-cured 6,60 114 / Air-cured 1.16 81 0,64 S2 Petique Stem 6,41 98 j Retcnrtitrted to~acca i 4,35 71 cigar 1.90 90 1.13 1 33 Flue 'k arbitrary unit of dye reducing activity her been defined ds that amount Hnidh lR~ngs ahout an absorbance ehanp of i.a per minute It 600 nm at annien; termeratvre (20·22't) in II an lirlht path at an initial concentration of 1 r 10'5, OtIP and pH R,E (d], BATCo document for Legal Services : Health Canada 20 May 1999 ·20· In general the cartelatien is fair, The outstanding deviants are perique tobacca, whore reducing activity is unuruaily Iru, and cigars. Our ~lultr ~ this point indiea~ that I~~e vapour phase is an efficient inhibitor of vinyl aatate polyn~riratiw~ and that lonju~ated Ciener, chiefly iro~nnt, alp pmbably nrpwllible far this activity. The ~artieulate matter prerents a mere canolex pictun, It is a relatively rtmng inhibitor and we find that snakes fmm differen: t~bac~a typer and inoiirg vehicle ase ligniiirantly different activities. The test rihi a we ale iludyini] is balically a aasure of the ability of smoke to intercept free ·adicalr famed during vinyl pol~a~riration and thus disrupt the chain prPcerr, eaoriderati0n of the eclanira of radical interaption and the typa of tomo~mdi knam ta retard poiynuriratim, rloger:r bat all three Redaniln~, additien, aton transfer and irec radical cwaling, contrilute to the inhilitary action oi sale, Because of the tomiatian of reducing adivityl as *nured Ly dye reduction, and innibitiol oi oolvmrilation, if is likely that hydmgn tranrfe, is the dominant Reci~anism. BATCo document for Legal Services : Health Canada 20 May 1999 _C ·21· REFERENCES i, Busse, R,F,, TobaecoChc~, Res~ Durhdm, N.G., 1967 2. Benedict, R.C, and Stedman, R,L,, Tobacco Sei., ~ 166, 1969 3. Benedi ct, R,C,, and Stedman, R.L., Experi enti a, 24, 1205, 1968 a. Bili~ria, M.H, and Nisbet, M.A,, Beitrage 1ur Tahtk., 6_, 27, 1971 5. Bilil3ria, M.H, and Nisbet, H.A., Beitraoe lur Tabdk,, 6, 32, 1971 6. Nisbet, M.A., Johnscn, 3. and Scnneller, S,, T6bacro Sci., in press I. 81um, H.F., Physiol. Rev., 2, 23, 1932 8. Georgieff, K.K., Science, 173, 537, 1971 9. re,,9ieff, K,K,, 81aikie, K,G, and Yhite, R.C., J. Rppl. Polyter Sci,, ~, 889, 1964 10. Elmenhor;t, H, and Schulr, Q,, Beitrape Zut Tabak,, i, 90, 1968 11. DontenHi 11, W,, Elmenhorst, H,, HalJle, H,P,, Aectteh, G,, Weber, K,H., Misfeld, 3. and Tirm, J,, 2, Ktebforseh, 73, 305, 1970 12. Elwnhorst, H, and Eri rmer, 6,, 2. Krebfors ch., 71, 66, 1968 13. toulson, C.A., J. Chern. Sbe,, 1135, 1955 14, SHare, M., J. 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