J)evd lllll ilelltofaQuantitativeMetho d forthe Analysisof NeutralSacc har ides in Wood Pulpusing Analytical Pyrolysis
by
JohnKell)', B.Se.,G.R.S.C.
Submlttcdinpartial Iulflllu.cnt oflirerequirements furthedeg reeof MasterofScience.
MemorialUniversityofNewfoundland 51.John' s,Newfoundland,Canada
Jul)'. I9I12
. ....
OfCanadaNalionalL,blaly 8'bliOtlv'1Qucn<lI'Onalc du Canada AcquiSilionsand Directionoos acquisoll(){\!i01 BibliOg,'ophicServiccsBra nch desservicesbibliOgraphi('lll{'$395WeUinglon Sl,OOI 395. ,ucW elIongtoo
~~fN""" ~'~ctitO<llill"'\
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a
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I59N 0-31::.i~7812 4-6
Canada
·ii- AlISTRACT
Trnd ttlunalmcthod sfor urcuclcnnilmtill\luftilenculT;11s'l~l:ha ridcCtllllpusilillll lll"Cll1l\I.I \· ~ blomarcdars suctraswoodPUlpM Cdiffi cull:ll\{l limc .ct>l l.~Ulll illt: .lJ)'comparison:Illal y lk :llllyn .l)'s!s techniqu esoffertheadvantagesof speedamlshupllclry.UndertheilPlm'pri:llcCtllll li l inll~,py",l ysi .~
yields protiucts lhatrctai n llles lercocon ligumlill1luf lhCpal\'ll llicutrals: lcch:lride Uhe a nll)'llmsugan;), The objective oflhis lnvcsngu nonW:L~to de velop amethodtuqunntiratclheucrnralsaccharidesin wood pulpusinglin-linepyntlysis - gas chnllnalngra phy(pY-Gel. lnilialeni 1l1.~ hIl: U.~Sl· lIlin lIIa:dmi zing the yichls ofnnhytlmsug:ln;
rmm
iSlll:ltcdl'lolysacch arh1cs rc."Clllhlillg lhm eflllJl\ll III wood.Faclun;suchaspym lysistemperatureanticauon-cxchuugcwell: invcsrlgatcd. Ncxt. n v:lricly or woo d pulps ofditTeringCIUIlpl.ISililll1:utdurigin wereinveslig:ltcdbypy.n c.ThenUI\Wll>IlS pyrnly ...tares producedby nescpUlps were cbnmctcrlzcdII)'Py-C C-MSusin gbuthctccmmhlljlaCI (El)nndchcrnlcnlillllisalion(ell.Fhl:llly. Py.GCmclhllllsweredcvclup,:dIII quanlify thes;\cchmilk compllsiliOl\s Ill' me pulps. C()IlIP:l riSlll l.~werem.ufcwith resullslI!lI:liIlCdhy :lCid Ilyd nllysis• t!eriva!i/,alil1nIgus chnun:l1l1grllphy. 'Twuon-ltn c pymlY/.cn;.lheCPS I'Ylujlnl hc 120'" amllll e:-;(;1:PYI\Jjcclll~.werecompared us tothclr SUililhililyjurquruaitntivcanalysis.
·iii·
m:DICATION
This thesisis detlicaled In mywifeAnnandIU my parents,vincent :lmlMary Kelly.
ACKN()WLEUli E1\IENTS
Iwish III expressIllyepprcoauon10 mySUllo.'rvhlr,.Dr.nnhIlclkur lind Dr.Mk h;ll'1 Mackey .fortbcircncou mgcmcnt.guidanceandndvlrc.1wIlIII.1alsolikeluthunkDr.HilmI('n'.:", )' andMs.M:1I1nnl3agg.~furmclrhclpWilhtheCbcmicalhmizatlon f\b ....'iSpccmunrtry.Fin:lll)·,IIIl.' financiulsuppn!1(Ifthe Schoul ofGral!u:Ile Slllllks illuc t'nnnnf :, fellowship.:u1I1nl Ill,' Dcpnrtmcmof Chcm islry,is grJlcfully acJmuwlcd gell.
GLOSSARY
A.II. AcidlIyd rul )'.~i s-ncrtvanxauonIOas Chromatography
CI Chemicalhmi1.alioll
CIMS Ctcuucat lonizalionMass Spectrometry
C.V. Cocnicicnll1fVari,lli(ln
DP" Degree of P(1)ymeri1.3titm El Elct:lmnImp:1l1I(1I\17.atiull
ElMS ElcctrunImp;.' lronszauon MassSpectrometry
EIC ExtractedInnChromatogram
rID Flamekutizatlou MassSpcctrumctry
GC
oas
Olwmalllgmphy111'l. e Ili);hPcrrornuncc LiquidChmmatography
MC Mit:mcry.~l'llllncCellulose
N.S.I'. Nllr:mtlaSulfitePulp
I'A PnmmAmnit)'
J>y P)' l\Ilys i.~
l'y·OC Pynllysis-O;.sChnlmalUgmphy Py-MS Pynl)'Sis-Mass Spectrometry TFA TrinullnJ:Il,."ClicAcid
·~i·
HilLE OFCONTI~ N TS
Abslmcl..
Giossary...
CHAPTERI:INTRODUCTION..
1.J.Pyrolysis -GiL~Chn.JlIlillllgf:lphy..
1.1.1.AnalylicalPymlysis.. . 1.1.2. Typcs uf Pyn,lyl Cr,;..
I.I .J.F;\ClUrsAITcclingl')'wlysis..
1.1.4.Arrlic,lliunsorAn:llylil'llll'ymly.~i s..
1.2.Woud Pulpx....
1.2.1.CUlllpUSiliuIlUfWuut!..
1.2.2.Differencesin ClenucnlCllmpll.silil)l1 llCIWe~·1III,mlwlklll_~
(AngillspcrlllsJandSullwolKls«(iYlIllMlspefllIS)...
1.2.3.The I'ulpingOl"WOlld...
I.J.TradilimmlMelhudsIll'Wuml l'u1rC:U1l\lhydralc Allalysis..
1.3.1.Nun-SredlleS:ll1:haridcAnaly.sis..
1.3.2.MOllllsaccharideAll:l lys is...
1.4. Pymlysis(lrCarhuhylll'lllCs...
1.4. 1.C;ulxlhydrdlcl'yrtJl y.~ i.~Mcchill\isms..
1.4.2.Malrixlnllucnecs ...
1.4.3.TIleSpccilidlyofAnhydm.~u~ ars..
1.5.OhjcclivcstlfthisSIllily
"
n 1.1 If, If, l2 22
"
-vii- CIIA l' rER2: EXPERI MENTA L..
2.1.Marcrlats...
2.1.1.Cheillicar.~...
2.1.2.Carbnhydralesand WoodPulps..
2.2.lIydmJysisIAJdilnlAt'CllllcDcnlval]zlilioll..
2.2.1.r.~ol lllet.lPolysaccharides ..
2.2.2.WoodPulps ...
2,).(j(;Analysis url)crivlllives... 2.4.Smllple I'rcp:lr.t1ion furPyRllys]s.... .
2.4.1.llo:louiz.llliullo rJ.~ola ictlPulysllccharilles..
2.4.2. 1' '' 11l11\E~ch;1I1geIll'WundPull's... 2.4.3.pI IAlIju.sllllclllufDciunlscrlwoodPulps..
2.4.4.Calil~lExchange ofSuturePulps....
2.~.l),...~cripl illllufI')'mlyz.crs....
2.5.1.CUSl'yn1jlmhe120 ..
25.2. SGEI')'mjcclur ...
2.(1.I')'ml)'sis -GllSChnlll1llln,g raphy..
2.7.I'YRllysis.G:lSChf\\ll\:llllgr.lphy.MllSS Spcclmmclry...
2.1.1.Ell·clrullllll ll;lcl luniz.minll... 2.7.1.Cllcmica! IOlliz.lllillll..
2.1'\. lnurgank Amllysis..
2.N.I.Ashing .
2..'1.2. The Dctcr minnionufIronin theAcid·W ashcll. Fch•
26 26 26 26 27 27
"
29
MJ 3J JJ JJ 32 32
:w
36 36 36 37 37 37
2.IU. SulfurCllllll'nlAn:II)'sis(Ifme Sulfite l'ulpsIt)'X-Kay Fluore scence..
CIlAI'TER3:\'YROLYSIS OFISOLATEDI'OL\'SACCII,\RlDES ... 411
3.I. GIUl:.ms... ·Ill
3.1.1.Mit.:nll:rysl:llIillc Ccllul(lsc .. -10
3.1.2.Amylnse.. 4,\
3.2. I-!C1CllljJIllysacd l'lrides.. .I.~
3.2.1.UIt:USIucnuGum; .I.~
12.2.Arahillll~:11:\CI:1I1.. ·17
3.2.3.D-Xyl'lll.. .rx
CI/AI'l' ER4:I'YROLYSIS OF \\,1001)PULl'S.. ~J
4.1.Inlrodllclinn ... ~I
4.2.Inlluencc ul"Dc;lshing .. SI
<1.2.1. llI'lt.:kSpruceMcchanicaJ1'11111.. Sl
<1.2.2.HlrchMcchanh'nlI'uill.. .~-I
4.2.3.BlackSpruce Kmli1'u1]1.... . .H
4.2.4.All'CU"'Bin:hOrg:\llusu lv I'lllp.. 511 4.3.Rcint J1 l( rll dJl~McwlCal i(lr\~•Tile EJ'lccl1I11 1 'ymr ys i.~... .~ "I 4.4.TheInfluence of Suspe nsion pllonmePyrolysis IIfWlltll!I'ulrl.~.. Yl 4.5.IIlcnlil k;ltillll uf l!lel'yl\llyt:l ICs lll"Acill·w' lshcd llhlck SPIU":C
andBin:11Mc..:hanicalPulllS hyl'y-GC.MS..
4.5.1.mcctronImpactM'lsS SfX.'Cl muletry..
4.5.2.Chemical hlilizaliullM,l~SSlll.-ctnunc try..
-he- CIIAI'rER 5:PYROL YSIS OF SULFITETREATEDPULPS...
5.1.hunuluctiun ...
5.2.The Effectorlun·exchangewithFet> . 5.3.ComparauvcYieldsuf AnhydrosugaTIlObtainedfmmaSulflrc Pulp
exchanyedwith aVariet.vofcations ....
18 7R IlO
82
5.4. Py-GC.MS (EI)ofSoflwoudSulfitePulps.. ... 84
5.4. 1. I'y-GC ·M S(13.1)of UntreatedSpruceSuturePulp.. 114 5.4.2. j'y·GC.MS (EI)IlfAcid-WllShcd SpruceSulthe Pulp... ... 86 5.4.3.!'y-GC·MS(EI)ufAdd-Wlished.Fel··Exchangct!Spruce
SulfitePulp..
CI IAI'I H{£I:VUANTITATIVI3.ANAl.YSISOF SACCIIARIDESIN WOOD I'ULI'SBYPY·GC...
h.l.hunuluctiou..
(\2.Ql1anlil1cmillllul" Glucose,IUd XyloseillaNumberufWuudPUlps II)'anExlem:1IStamfnrd Meehud ..
fJ.2.1.OptimizlLlint1ofthe Pyrtllysiscondtnons...
/1.2.2. QU:lIllilic:llilintil GIl/eliseAmiXyjosc ..
(d.<.)lIanli"I!ivc AnalysisIll' Ihe MajLlrSaccharidesin F!i ' .Exchanyed,
!'iulli h:-T rcalctlPulpsusingaSlmlllllnljzcdSulfitePulp...
h.~.r.Qunnlilka lillllusinythe Pynlp rohc"' h.J.::!.QU:ll1llficalillll usinythePyrojeclnJ"l 004.QU:1Il1it:llivcAnalysis(If;tllNCUIf:d Saccturt ucsin the Ackl-
W:lshcd.Nou-Sutthc PulpsIIsil1~:1Stnndnnlizcd Pulp
rlIAI'1' ER7:CONCLUSION .
ss
9S96 96 1(11
101 lUI
It .
110 liS
-xi- LISTOFF1GUR":S
1.1.Tilt SlruclurcufCellulose .•.•••~~HM_·. . _· . ._~. .• . . _ .. _·H.._ H · _ · ·_ .._.
1.2.Tilt Structuresofthe"bjurIlcmicclluloscs 3)Ambinog:d:JClan (Sofiwood) alklb) GalaclugluC(lm:llVlan (Soflwood).. .._ __ __ _•._•.•__.
1.3.TiltUgnillIm'Ut'MIl"!i __ ._••_ _.._ _•••~.._ _ __ _ 1.4.Ex:unpleuf the Lignin DcgradalionMechanisms in Kn fiPulping (53) 10 I.S.Exampleof theLigninDcgrJ d,lliunMedmnislllSill Sulfilc Pulping(54) II I.fl.AldilulAcctatepcrivntlzalon.COllven;ionof n-otu cosc Inlo Glucilul
Hcxacctatc
1.7.HemolyticFisslullorCenutcxc(72)
1.11.IlclcnllyiicMs.~jun,,[ Celluluse (7]) _ ..
I.IJ.I'ymlyiicl-';athway [ur liltFunnalioo of1.6-Allhydrogluco[unr.osc (74).__ . 1.lll Fun n:l1itlll ll[Nun-Sp:cilicCartdlyUr:ttc PyroIYL:lICS(71)_.•.•_ _ _ .
15
"
19 20 21
1.11.l11CSal,:harida;[lIuI.1in Wllod ;11111ueAnhytlrosugars lheyronn upt..
I'}'n ~r-'i ~ M _ M _ _ ~ _ _ __ 23
2.1.CDSl'yn~W"I'I~120·''' H _ _ __ 3]
_~.SGEl'ynljl'Clllr" _ _ _ _ _ _.. 35
ll.l'ynlgr.ullSnfMicnlCry!(fallincCellulose(a)Aslllcssand(b)'treated withll.ll:i MKO...
.1.2.I'ymgrams I\r AIU}'ltisc(a)Unlrc,llcdand(11)Acid-Waslll:ll...
JJ.l'yn1gmmstil'Locust
ncm
GUlli(a) Untrcurcdand(11) Pmton Bxchangcrl .lA.I'ymgr.llllsIll'1'nllnnE.~cJ~\lIl:! ~dAmllinogalaCI:lll...
41 44 46 48
-xil-
3.5.Pyrogrums ofXylanrromOal Spells ('I)Uillremcllmil!tb}r'nuuu Exchanged...
4.1.Pymgrnrnsor Black Spruce MechanicalPulp(a)Unlrc:LINImill(h)
Acid-Washed .
·N
4.2. PymgrumsofBirchMcchanlcnlPulp(a) Untrc:llcdand(h)/\ dd· WashCll .. ~.l 4.3.Pyrogr.lIllsof BlackSpruceKmfi Pulp(a)Untreatedami(h)Acill-W;t~II~'c1 ... .~~
4.4. Pymgrumsor Birch Ak'ellillPIJlp(a) umrcncn.lIld(b)Acid-W;IShcd... ~rl 4.5.PyragrarnsIll' Acid-Washed mackSpruceKr.'ft Pull'Treutcrl wilh(:I)0.1N
KCI.(h)0.1NC:LCI~:nul(c) 0.1Nreo,
4.6.TheInnucnccofSuspcnslnnpHun theAnhytlmsugarYlchlsfurBI;u.;k SpruceMechanical (\')alld Kran(0 ) Pulps..
4.7.TheinnucnccIll' SuspensionpI!on IIle AlIhytlmsug,lrYiel.lslilr Avice! (0) andee-Ccllulusc(0)..
4.8. Pymgrams01" Spruce Milled WotldLignin(a)Uutrcarctmill(h) Acid-W'l\herl ..
4.9. RepresentativeElectronImpact(EI) Speumof the AnhyllllIsugars,(aj1,(1- AlIhydruhexullyranusc, (b)1,(I·Anhydruhexofuranusc,(e)IA-!\llhydm- pcnropymnosc.and(d) lhcEISpectrum uf l,4-Anhydrog:llaclnllyr.1IIl1.\C..
4.10.Total InnChnnnatogmm(TIC)for Aeid·W:lshed1I1:1C\( SpruceMedmlliC:11 Pulp Obralncdby Py·GC·MS(EJ)...
4.1J. Tn1311ol\Cnrmuatogmm (TIC)forachl-waxltcdBlrchMedwnic:lIl'lIl11 ObtainedbyPy·GC·MS(Cl)...
4.12,Olcmical loniz,ltion(Cl) Mass SI'CClrJufLcvegfucnsuuU.~illgdifferent Reagcllt GOlscs (3)M cthane, (b) Il-lso bulanc aJld Cc)AIlIIlIOlli;1 .
.'is
f"
f"
-xill-
4.13.Trualhill Chromatogram(TIC) forAcid-WashedBlack Spruce Mechanical
4.14.'rulal lollQmllllatogrmllenC)for Acid-WashcdBirch McchancatPulp Ohtainedbyl'y-GC-MS(NIl,-CI)..
4.15. Ammoni,lOlclllieallonil lilion MnssSpectraof (a)4-YinylGuaiacol
:lI1d(h)4·MelhylSyringol...
5.1.Pymgmmsof Aspen SteamExplosion Polp(a) Untrealed,(b)Acid- Washedand(elFe: '·Exclmngcd•.••
15
76
77
79
5.2.t'yrogmmofAvicclMicmcryslallilleCellulosesorbed wilhDilute HlSO... 80 5.1 I'ymgmll\.~ofNlll'and:lSullilcPulp(#23J()l)()(a) Acld-WashedamI(b)
Fcl'_Exchangcd..
5.4.TnllillullCnromnugnnu(TIC)ofuntreated NotandaSpruceSulfite Pulp obtalncdhyPy.c;C-MS(EI)...
5.5.TUI,IIlUll Chnxn.uogmm(fIC) ofAcid-Washed Ncrunda SpruceSulfite I'ulpohillinetlhyl'y-GC-MS(EI) ...
5,(1.TileEIM;\s.~SpectralD,ltnmill Pmposetl Structuresrorlhe Unknown Ligninl'y m I Y/~ltcsSlI. 72nnd81l ....
5.7.Tutnlron Olwllla:ugr:nll(TIC)forFe1+-Exch:lr1g~'(1Norand;lSpruce
SullileI'utp ubtalncd hyPy.GC-MS(EI) .
fI.1.
'rnc
Eflh:lufPymlyxix Tcmpcnuurc onrncAnhydmsug<lrYieldsfor Glucose(0)and Xykrsc(v)InunAcid-WllshedBlackSpruce l\kehlmic:lll'ul p•.•6.2.Linear DynamicRange Curvesfor Anhydrosugar Prouucuon forAcid- washed 1lI:\l:kSpruceMcrtunical Pulp(:1) Auhydmglucuscsand (b)
81
85
91
94
97
·xh··
1,4. Anhydmxylopyanosc .
6.3.S';U1d:lrd PlOlSor AnhydnlsugarYield versusSampleWl.'ig!l1furlol)the Anhydmg!ucosesfromAvice!MlcmcrysudllncCcuutoscand {hl 1.4.Anhydroxy[op}·mnoscIrt uuDctonlscdXyl;mrmlll OalSl'dlS..
6"'. SampleWeigh'versosAnhytlrosugarPe;l\;'Arc:lforAvicelMicnK'r}'~1 ;llH llc Celluloseu~inl:thePymjeclor'"...
6.5. zxtractcr1011Ouomctogram(mIL57)fllfAd d·Washedm"r kSprurc MechanicalPulp...
!W
Ill!
110
II'}
I.IST m'TAULES
3.1.IdenlillcutillllortheImportan t CarbohydratepyrulyaatcsObservedin Ihel'ynJgr:ulIsuf lhelsnlarcdl'lllysacch :trides....
3.2.Yieldsof 111CGIUCfl.'OC Anhydmsugal5formeGIUeall~...•
4.1.IdelliineatiullormePcuksinthePY-GC-MS(EI andCI)1'01011Ion Chrnmafngrams forBl:lekSpruce(Acid-Wa.~hed)andBirch (Ad d· Washed)...
5.1.CtllllpamlivcYieldsofAnhydmsuJ;:II'SIrumCUI IOI1-Ey. ~l1angedNorunda SullilC Pulps..
5.2.IdeulincliliunofthePyrulyzules intheSulfitePulpTIC's....
rd.Opril1liz:Jlinnnfl'yrnly.sisPur.llllclers..
(1.2.I'Cf\.'C1l1agcGIII\.'(lse alldXylnscCo1l1 P(l~i1i(l1lofAcid-W:LShcdPulpsas DClcnllincdhyAddlIytlrulysis(A.If.)andPyrulysis·GlLs Chnllll:L1\l);rUllhy (I'Y)..
(l.} .t'crccmngcS:lccharidecomrostnon(IfFe:'-ExehnngcdPulpsasdctcrmtncd hyAcidIIr drulysislDcriv:Ll izalinn.GC (A.lI.) and Pyrolysis-Gas Cllrllmaltll)raphy (rY)..
/lA.I'crccmal)c S:lech:mdcCompos lnonor Fe:'-E xehlLllgedPulpsObmmcd
\lsill~theI'yn\jcl:l\l~ .
(1.5.TIl('I'c rcc nmgcClllLlPIlSiliollfoeauuc Saccharide sin(lieAdd- Washed PUlpsusingAehl-W:lsIIlYJR1:LCkSpruceMechanicalPulp asaComparartvc Slaml:ml...
42 43
69
8J
"
OS
101
1m
I())
112
-svl-
6.6.TilePc rccl'llJj;e XylllSCmmcA,'hl·W:lslll,' lll'u!ps Ilclc'n1\illl'dlIy(h\.'
methodor KlccnctAI.(96}••.•.,.,...•.•..._ 11.\
CIIAPT F.R I:INTROD UCTION
1.1. 1')'TuIY.'lh·(;a_~ l1tromaltll:raphy
1. 1.1.An a lyticalr)'rot,u~
Pymlysi~canbedescribed as a~mjcaldq:mla lionreactionlhalis induced bylhcnn~
energyatone(I).An:I.'yli~1pymlysbcanbe furtherdellnedastheeh3raclcri 1.at iooofam~ri:LIor a chemicalpmccs...bytheInSlrumefllaJ analysisof it'spyro lysisprodLK1S(I).
Though thelirst sden li llcuseof analyticalpymlys iswas docume nted in 1860 (2).ilwasnOI until the1941l'stllill pYlldysiswas uxcd in combin ationwithmodemanalyticalins!nUllCntalion.At lhi.~limeitllUlllhcrof reportswerema de oftnc useofrum spc ctromelry to characterizeuic pyn llY/alcsofsynlhe tlcJllllymc rs (3,4,5) .In 195 2, zemanyusedpyrolysis.mas s srcctromclly(Py- MS)10dla r.M.1Cri,.cla rgemolccul:u-weightbiopolymmsuchasalbuminand pepsin(6).Hisresults were signinC:IIll inlhatchJrdCIeristic pyrolysispro lllescould be rcprodllciblyobtainedfor these CClll1pnuntls. TheIlrsllrud yinlcgrated pyrolysis- mmspcctrceetricinstrumentwasdescribedin 1953 (7).
TheilllRk!\K:li onofgas-liquidchro~og r.Jphy(G C)In1952(8)rapidlyled tothenrst repu nsof it'suseinCtll11hill3lionwithpyrolysis (9.10.11).Pyrolysis.gM chromalography(Py-GC) 4uicklyllvc rs ha!klwedpyll.,lysis•nlass spectrometry asIhete clmlqac orchoIccduetoit'sabilhyto scpncnevnl;llilcpyrnly;r.~le.~priorInucrccu on,and becausepyrolYl.crscouldbeeasilyinleg rnledwith GCInstmmcrus ,Inadc.l i!illil.m;L~ Sspcctromeuywas vcryellpc nslvcand difficultto usc.Then:cent ucvctopncmur ruggedoJlCnlubularca(lill:l.I)'columns&rcaUycntunccdthe1CS01v ingpowerofgas Chrtllll.:ltlla;rap hy• .:Ind bassimp li fiedthe interfacingorGC10massspccnorneers, Cclllp!ex
.z.
pyrolY7.3lC mixtures cannow he resolvedaooummlhigouslyid~\Ili lic dhyJlyn~lysis'~ilp i l laryCIllu llill gas chromalography•massspccrmmctry(Py.G C· MSl.
The techniq ue ofPy-GC. in allus jonus,connuc sIIIheIheIIl~lIill;lIltmctluujillld ~ll)'lical pyrolysis.However,Py·MSIsu stcadu y itx:reascdinp.lpul: uilyhcc:lUSCll(it's:Ihl!ilyIIItkl('(11 ;1 r~t,'r molecula rweightpyrolyztllcS11marcunableto Pil'lSlhmugh;lGC11J1l11Un.l11t,Il"v dtlplllcnt s in analyticalpyrolysis ill.'i!rumcnlmlol1have been reviewedonanumbernf occustons(12 , 13,17.1N).
J.U.Types uf Pyrolyter s
The majorityof pymlyl.crsused in Ctllllbin;llillnwit hGCfallllllntlll'CcC;llcl:urics;rc.IISlivcly healedruemcrss.ln ductlvclyhealed(Curie- puint)lillllllcnts, llllllclIlltillll\luslyhC ;~Cll llli c rtlrum ac cs.
The nrst twocaleg(Jric.~arcalsoc1a.xsifiedas pulsePYCllIYler.~. lr:lcll(Ifllie ahuv..: IYllI.'); was devd opcd earlyinthe cvotuucnof analyncalpy ml ysi.~andtheh:l.~icfeaturesnavedWllj,'Cdlillie sincc(l4.IS, I6).
Resis!lvcly healedpymlyzcrs arc usuallycomposedOrjll:IIiIl UIIlJilamclllsIIfrihhous.SlIlu h le samples canbeeasilyapplie d 10the metal surface,An electriccurrculs Ihcll1J:l~...ed IhAKlghtile wire,wbichInlum heau andpym[YI.esthe sample.Insoluh lesamplesC:UIbepynllY/,cdhylll:lci l ig them In thecearc ofacoiledliIamcniwilhthe aid of a quartzhoat ortube. Mndcrnrcsisuvcty healedpyrolyses.sucha.~the Pyruprobc'&(Fig.2.1)aOOthePymra~.an:dcsigncillutighl lyt~llll n ll impor1 antparamete rssuch asthe finalpyrolysisle mper:rturc andtcmpcmur crise lime.
Inductivelyheated pyrolYlers operateon a dif1ercn tprincip le.Asampleisap pliedtuIhe surfaceofa fcrrmuagnetlcwlre whichisthenplacedln Ihe ccmrc01"ahighrreq ucllcyind uctirxlcull .
-3 -
Am~gneticf1u~i.~Intt fuicd. which intum inducescddy currcntsin thewire'ssurface.Th e
ru emcm
ishc~r ctlra pi lllyunlilatemperat ure(Curiepoin!)isreachedatwhichthewlrebecomesparamagnetic andabsorb snuIll(JrCenergy.11listemperaturewillbcmainl.aincdaslongasthe"field isapplied tolhewire . TheCuriepointCWlbeva ried fromapproxima tely 30010IIOCI'Cbyalte ringth e Icrromagnct tc alln)'co mrosmon ofthetilamCllI.Forinsolub lesamples,boats,tubes. and v-shapcd rihhullscanbeulCd. Ccrle-pointpyrolY7-Crsarcch aracterisedbya vel')'rapidtemperature rise lillle hu larcInnexihlein that theIln alempcruturcIs dctcnnlncdbythe compositionoruewire.
MtcroremaccPYTlllyzcrs arcsimplerindesignlh;m!he previoustwocatcgortcs.Theyusuall y
consist ufa qunnztubcsurroundedbya tcrnpcmturc -regulatcdmlcrcrumacc.Thetube isconsrcnuy liwe pll1yan inen gas(usu~ lI ytheearnergasforthe Ge). Samplesareeith e r droppedImctile
py ro lysiszone(vcnic al(unlOlCc),orarepl acedinaboatwhichisthen1lI0VOOinto Uteheatedregion (huriJ.lllllal rumacc). Solublesamplescan alsobe coaledonawirewhich isthen harouuccdlmc thepymlyzcr.Thoughmkrofumace pyrcl yzers can give pyrolysisprofilesrlmttario noseobtatned ustngepulsepymlyl cr.Irreprod ucibilitycanD::aseri ousproblem.Thisismostoftenduc10 slow lcmpcmturerise times andseco nd aryreactionsoccurringastilepyrclyzatcsarcsweptUuo ugll lhc large hCOllctiZOIlC.
1.1-1.recto rs AlTectingPyrolysis
11lCrcarclIl:lnyparametersthatmustbetlghllycootrollcdIn orde rtoguaranteereproducible pyrolysis.M;my(Ifthesepnmmcrcrshavebeenextensi velydocumented(13.19,20 .21).IIisagr~'Cd thill ;1p>'nllyzcrmustbe able lllrcprnduecU1CS;Ulletcmpcmurc-timc profileand that, inmost cases.
aICIIlJlCrnlUn: rtscumc inII~orucroruumscconasis mostdcst rablc,Inaddition,samples mustbe
. ..
homogenous.StIl3l1lnma.u (5-100111;).andhave alalJcscrraceIIIYO! UlhC1".-11Il,n. nrli alll.l'..·hh theserecommcnd:lllOfl.'lshouldmlnimlze theprohlcmlloflhcn nal 'f:ltliclll';IIllJSC\lllIbry n'a..1itlll"
occurringwithin thesample.
Otherradars an:im(lOnanl inPy.ocForCll3Illri C.a I,il:hearnerga... vrltllilywilililltill.' pymlyl.t risrequired10c:n.wrc the(astremo valofVOI;tlilcryRlly / illCOl(R,mIIIl;:pyml)'l;i,711111.:.l1lill muslhedone inanlcrtorfCV'Cnlscton d ary re;lctlo n,whichluwer lherichls uf IIICprim; lfy.hil:lI musspmdocts.However.Hthcg;unow is100r:L~1illl1~yancel lhercmpcraturc-umcororuc.Clllll spots!lclWCCllthepyrolysiswileamIlhc GC columnare m\l)lhc rp1~ Cllli;t1problem.'11ICrc~illlls111 betw eentheseIWO l<HlCSmU~1bekC(llsufficic mlywan utn orderIn preventtheCIlI1([CILs ,llinlluf lhe highermolecula rwciClllpyruly7.:aICs.
Analyt icalpyrolysis(1If1lin ue:'lIUliuffcr CnHIl lhcmililakolhclidIh;llilisirn"f'Rltlucihlc.II hasbeendcmo nslDlCtllIlalCYCIlaCOOlinuouslyhealedmicnl(um :ll:c(:InrepRldocc pynIlYI,OIICl'1lins forsyntheticco- polymenlntbc:creeecr 0.45'1>n:I:1IivcJilnlanldcvi;llMHI(22).Re~u ll~Midi;r'\lhis dcmonslr:J[ethatIhighdegreeorprecisionC3I1be:Il,:hievedpnlvltlcdthepyrolysispnw.."C\S is opIimiscdandrigidlycon trolled
However.interlaboratoryrcproducibilUyha... hccna ...erinu~pmhlc mwithhUlh1'y. (iCat.1 PY-MS.Un fOrl unalclythetlcvclopmcntoranalyticalpynll)'si,sh;~snotbeenwellcllllnlillatc tl. '11lis has resuhcd in alargevariel)'of lnstrumentxand pracuces.CompanulveinlCrlalxlnl hll")'lria lshave beencarried outforbothPy..oC123-2H)andl'y· MS(2'.1), Initialqualitativeandquamlmivc comparison'!forpy..ocwen:[IOOr.especially for CUlllplC.r1n1Y1llCIlI(23 -25).IIl1wcvcr. the ta«
trialsscistnctc rguideline sforallSICp!!inthe:maIysisand thisrcSlJllcdinsignilit::UllilUrHHV CUlCld.S
·s ·
in interlaboratory prec is io n forbothpeakInten sityand retentionlime(28). [Icanbe conclud e d thai aslung as lhe variablesarc controlled,differen tpymly zcr syste mscan produce similarresuns.
1.1.4. Applicalion~orAnlllytil:al Pyrolysis
Organic polyme rs,wtlclhcr syntheticorbiological.arcdiffic ult [0analyze by more convcnli on:llanalylicallcd miquc s becauseoftheir poor solubilitynnd/orcomp lex ity,However, pymlysisbreak suown pulymc rmolcculcs[010 smalle r. more eas ilyIdcmifi ab lc fragm entswhichare
frequentlychnracicrlsttcofueparentmolecule.Itisin thisareathai analyticalpyrolysishas made u'smust signincantcomrtteue ns.Inaddition to mosegiven below ,therearcmany examples orne dfffcrcnrap(llicaliol1s of analyticalpymlysistoIJcfound in theJournal ofAnalyticaland Applied PYl1Jl y.~i.~. Purrhcnuorc ,anumber or comprehensivereviewlexlsandarticlesarcavailable (13.30.31.32),
Analylic alpymlysish:1SImportantinduslrialandrorenstcapplfcationsInthecharacterization llr synlheliepolymersandrubber.>(33.34.35). II Isalso used in limanalysisorgeologicaland envinmtuenlal malerialssuch as coals(36).kerogens (37 ).soils (38),andsediments(39).Inrood chel1\i_~l ry ,uctccbnlquchasbeenused10lnvcstlgatethepyrolytie reactionsdial occurduring some cookillll proces se s (40). Analytical pyrolysishasalsobeen used todiITerenlialebetween micnlllrganisllls(4 1).Finally, ithasbeenused10characterisebiopolymerssuchasprorelns(42), PUIYS:LCclwillciit(43).li gnins(44). and, aswell asintactbiolog ic alsystemssuch aswood(45) and spruceneedles(46).
·6.
1.1.Wood Pulps 1.1.1.Compos ition or Wood
Wood isIhemajorsupportivetissueintreesami otherhighertinier pl:mK IIis:lhu responsibleformeupwardmovementof waterandnutrientsfnun Ihe nwllS10Ihe leaves:mu lIl'I",'r partsoflhcplan l. Fromachemicalpointorview, 90- lJ5%nfIJ~drywdghtof\'Iuu,1 can I""
accounted Ior by threepolymerclasses:cellulose, ncr uiceuulosc, andlignin(471.IntlrgiUlic ash 'Il.J enracuves.suchaswaxes.resins, pnncins. gums,etc.•makeup theremaindernftheIlrywctgju.
Cellulose IscompnsedcmlrclynflA-linkedB-D .glucopyr.lnosemkl ;LCcmllllsfur4n-50 ')f,uJ thedryweigh tor wood(Fig.1.1).IIis IIlinear pu lyme r withanumberaverage degree III"
polymerization(DP. )of 9,000•IO.(XJ() (4X). uccauscofII'Sabundance arH.!commcrcial huporumcc Itbasbeenextens ively invesligated(4XAlJ). TIleindividual moleculesarc:lggregOltct.! uuo mlcrnscopicstru cnsescalledmlcrolihrilsami theseformlheprimaryraneewurk Ill' meccnwalls (47).
Figure1.1:TheSmrcturc
o r
Ceuurosc.-7-
TIlehcmiccllul(JSc.~arc mlnuresof short chainpolysacc haridesCOp.=100.2(0). TIleYarc closelya.~s(Jd alctlwithcellulosc andligninanditisbelievedthatthey aCIesabin~ ingmatrix.for mcscIwuin.~(Jruhl cpolyrllcrs.Fully 25·35%uf drywoodishemicellul ose.The srrunurcsofue rnajnrsof!wucK!tcmiccuctosc..'areoullincdinFig.1.2.
Lignlns arcthre etlimens ionul, polyp hcnolicmacromole culesth a tarcdistributed throughout lhecell wallnnd lhemiddlelamella(intercellularregion),LigninisIncorpo ratedIntothe11I311during lhemalumliull "flhecellandgivesitslrcnglhandrigidlty (47) .'Ibcthreelignin precursorsarc listed inFig.1.3(4K),TIlemommcrtcunits arc randomlylinkedviaeucr and carbon- carbonbonds.
Thereforethe re isIXIIypical repealingunit as uere are for lhe polysacchari des.
1.2.2.lIi1Tcr c/ll'CsinChcmicalCo mpositio nBetweenIlard weeds(AngklspCrms)and SuRwouds((;y nlllU.~pCr nl.~).
Asa gencmlrule, lmrdwoodslindsonwoodsdifferIn(heirhemicelluloseandlignin CUlllPIlSlllllIlS.SenwoodncmtceuuloscIscommonlya 2:1 mtxiureofgalacloglucomannansand arahlnoglucunmox)'hUls.HardwuodhemicelluloseIslargclyeomposcdofglucuronnxylanswithsmall llU:lllti ticsof gluclInmnn:ms(4K.49),In addhlon, hardwoodxylansan::heavilyncerytnred atthe C2 nrC] ring flosirions.
SUrtW(101'Iigninsarederivedalmostentirelyfrom eonifcrylalco hol(guaiacol)precursors.
wllCreils hardwoodIIgninsalsocontains)'ringylpropaneunits(t.c.derived fromslnap)'lalcohol).
Dcriv:llivesof/l-L'(1um:lrylalcoholarcfoundinminoramountsInallUgnlns.
a)
b)
.K-
Figure1.2:TheStructures oftheMajorHcmiccllulcscs.a),ArnmnoglucumllOxyllln(Softwood) andb),Galectcgtuccmannan(SaRwood).
yH,OH CH
\I
¢
OH
p-CoumarylAlcohol Figure 1.3: TIleLigninPrecursors.
yH,OH CH
\I
¢tOCHl
OH
Conifcrylalcohol
CH20H
tH
\Ien
C HlO*OCHl OH
Sinapyl Alcohol
. ,.
1.2.J.The Pulp in;ur WOlKI
PulpingistheprflCCS5bywhichwoodis reducedto fibers.TIle choice ofpulpingprocess willdcpcntl lll1a numherof(actorsincludingtheendusc of!hepulp,the:treespecies IIIhe exploited.andthefdallve cess of!hevariousprocesses.1lJcmost widelyusedproccsscsf:lU imolwllc:llcgnrics:mechanicalantichemicalpulping.
Mcch;U1ical pulps arcohtainedby ahr.Jdingthenbcrs fromtile wood.TIdsisdoneeither by pressing debarked logs agahlsla revolvinggrindstoneorfceding wood chipsbetweentwo rotaling.
gnl(lvcd disks (50).Therenn erprocessnormallyyieldsapulpwithshort fibresand alotof lines.
'n lClatterpnlCCs.siscalledrelining,U1d produces amorehomogenouspulp.Thesepulps arcused In make pallerwitha highop;lcily.An importantad\l3f113gCof lhc mechanical pulping islhonthere isnntcwa.~l agc(i.e.•pdpyiclt1.~arc90%or grealer)andthereIsnotoxicdDucnl.However,these IIIccruPlicalpn~ ~irea large cncrgy inpuL Inaddition. papermadeeXclusivcly or mec:lun lc:al pulptearsea.~il yand yellows'1u1ck1ywtenexposed10light Generally.mechanicalpulpsundergo furthertreatmentand may be:IIlh:c:dwiththelonger fibresorchcmic31pulpinorder10strengttlcn thcfilial pmdul1(51).
Olelllicitlrulpinsprocc sscsreleasetilefibresbychemicallyattackingUleencrustinglignin plllyma.ThereliretW(Ipredominalltprocesses:krnfl andsulfileru1ping.In thekranprocesswood chipsarc usu:lllyconkedfer1·2hoursOIl 17o-18(J'Cin asolutionorNaOHand N:tzS(52).The pmpl.lsedlignint1cpulymcril.:ttiollreactions aredescribedIn Figure1,4 (5]). Krnflfibresarelong.
l!;ut alit!slnmg ;111\1arc usedtoproducepaperbagsantiboxes.Btcached kraftpulpisusedfor fine Il:lper.t1iS.·iUlvillJ:pUlpsforthepolymer indu.'«ry.andror whilepXkag iot! board.Inatypical
.10-
Cll~OH
I"
IIC- OR~
I .
~
,R,OIlyOClI
J°
1 ow,
SirfilOH
~. He,
0'Figure1.4:Exampleorthe LisninDegr:ll.lationMechanisms in KraftPulpini,:(5] ).
kraftpulp,80% orthellgnin. 50%orthe hemicellulose (especiallytheglucmnannansj.allliIlY;Il,nl thecelluloseareremovedduringcooking.In:u.ldiliun,the acetylgroupsandpcrKlantsidechaiu_~ul thetcnncellulcscsarcquickJylost.
.11.
1bcsnme processis.inIact,anumberofpulpingprocessesthaicoverthewholepHrange. hom~1J1(lJrdioxideMllulions(pH ,.1) to sodium sulfilCIsocJiwnhydroxidcsolutions(pH" 13).
Noonefex tiooscheme en describe !he lignindcpolymeriulion processesthaioccur duringcooking.
lhough thc.d phaClrtxlnofthepbl:nyl ptopal'leunit seems UI be:the siteofinilial~tInmost C:L'ieS.A possible n:::lClionsctereeforncul1:llscme(pH6-10)pulpingisdc:;aibcdinFigure 1.3 (54). At eithe rpHextremeligninremovalandhemicellulosedegt3cbtion iscx' ensivc.Xytans arc
"
CIl1
01lk b
rr~-o-<Q>-CII-!ll-Clr,orr
lit 61l
r{~,
r
110lt~o r=:" Q '
OCII) ~~Fis,:urc:1.5:E1I.3mpl':or Ill.:Lignin tXgr.llbliu nMcchanisms inNeutr:USUllilcPulping (54).
·12-
preferentially [OSI at low pH whereasthemannansare caster 10remove11\all;.'llineprocesses .Pulp yieldstend to behigher (75-90%)formid-rangeprocesseshut mccluuucal dcfnbationIll;!)'I~
neededto completelyseparatethe fibres.Because of thel1el lbiltyor thesuture!JRICCSS.pulpswilh a widevarietyof chamctcristtcsaod end uses can be pmduced.Forexample.Ol..:illsullih.'pulps[luw pH)arcused for dissolvingpulpsandlnrcnncdtac pHpulp.~(pI14- ltl) canhe uSl'tlfllr11CWS[Ullll, paper,and corrugated board.FinaJly,the recently develnped alkalinepmcess (pll 11-14)pmcluccs a pulpsimilar10kraft pulp bUIwilhout the 1l0000Ious odours a."st\Clalcllwilh till' Innerprocess15<1 ).
'ne rc arc many processesIhat do notlitne:ltlyinto eitheroftile :1I111vec:ltegllries.
t-or
elamplc,many processesuscaninitial chemicalso aktosoncnme H!!lIinpriur to grindingorrefining (cbcmimcchenlcal).Otherprocesses uscIlea:to achieve lhe same effect (lhennllmcch:mic:t1).Stcam·
explosionpulpisproducedwhen woodlhalbnsbeen treatedwhhS!C,Ull(wilh or whllllnichcmicaL,,) alhlgh pres."lIresIssuddcnly dcprcssuriscd,Finally,lherearetuc recently devcloped...oJvelllllllllling processes. A goodexam ple wouldbe tile Ak:eU'"ethyl akuhnlpulpingIlnlcess(55).Wlllldchips are cookedin analcotml-wacrmixtureut2OU'C.Dcligililic:llillilis enhanced byIhecollvcr.~hNIul the hemicelluloseacetylgroupsInro aceticacid.
1.3, Tr aditionalMethods of WoodPulpCarbohydruleAmlly.~i s 1,3.1.Nlln-Splocifie SaccharideAnlllysis
TIlefollowing are examplesof commonly usedmethodsfurcharactcrizlng Wtludand pUlps that do nordistinguish betweenindividual lllonosaccharil.les presentill lhe s:ullple. '111Ctnl;ll carbohydratecontent isucrcrnuncrby removingthelignin,ll1is canheachievedhytreaHng urc samplewithchlorine gas andz-amrnocmanoruntll a whileresidue,callednotoccnurosc.remains(.'irJ).
-13·
tllCvitah ly. some oflhchemicelluloscwill be losttluringthistreatment,
TIleccuulosccementcanbedctemunedinanumberof ways,For exam ple. Crossand Bevanccuumsc isnhtaincdbycntostnauonofthewoodsample. followedby washingwith2%
N~ S O land 3%SOl'and linallyby bolling in NalSO)solution(57).Alph a cellulose content Is dctcrnrlncd bytrcatlng holucclhllose with 17.5%NaOU (58),Thisprocedureremovesmost.though nnlall. of lhetcmrecuutoscs.
11lcpemusecontent.xyloseandarabinose. is determinedbyboilingthe samplein3.85 N IICIwithsome NaCt (59). These sugarsrcr m furfur al whichismensepara ted bydistillation.Its corcentrauonisuctcnutneucolurimctricallywithorcmct-rcc i,reagent,TIleurcnlc acidcementcan hedeterminedhygencr:lling.andsubsequentlyanalyzingfor.COl fromiL~carbo xylicccrdmoieties.
1111s isdun e by ;ln ilingIIlCsamplein 12%HCI(60),Alternatively,colorimetric analysis canbe used (61,(12).
TIIC wood polysaccharidesarcreduced \0theirconstituent mcnoeaccnartdcs byacid hydmlysis, Themust co mmon lyused methodis sacman hydrolysis(63). The woodsample is lrcall'tl wilh72')\,H~S04at]O'CIur I hour10 swellit. TIleacid Isthen diluted 10 I M and the suhuion isrcfluxcdfor2l\l5hours.Theinsolublematerial Il:m:tiningisreferred\0 asKtasonlignin.
even Ihl1ughitmay connun;L~muchas15%carbohydrate.
'rrmuoneccncacid (TFA)isalsoused\0hydrolyzewood and wood pulps (64,65,66).Itis
-14.
a vnluttleacidwhichfacilitatesit'sremovalafterII~hyd!\llysis step.In '1l.llIililln.,mhyllnlusTFt\
is a good solventfor cellulosewhichcan aid in h'shydnll)·sis. Fcngel ctnl.havedcvdolX'd ,I hydrolysisprocedure specifically forIignocd lulosi~s(65). TIlesampleisstmkcd tlvcmighlin anhydrousTFA at room tcmper aiure.followedby reOull.ingfor I hour.111C slllullllll ismen t1ilul,'d 10 80%TFAandrefluxedforISminutes.Finally the sofunonisdilull'd 10J(~,TFAatulrl'llu~ed for2hours.10 anomerinvestigation,POlice crat.hydmlyzcrlHgnecelluhudcs,uuplcs inutncr cnr corccntrattonsofTFAOIlIOIJ'CInscaled,evacuated vials (6fl).TIleyfmmd1I1,IIIheIleslresults were oblainedfor 2hours in110%TFA.
Ingeocmlthosemethodsarceffectiveat releasinglhcneutral1l\l lnnSaC~Ii;lrillc-sInnulh~
samples,However,llulcor nouronlc acidisdetected.IIInddhhrn,SUIllC(Ifthe1ll1ll 1l.Is;\C~ha ri lles
lireacidlabile(e.g..arabinoseamixylose),lIulcaremusthetakenlUllIIIdcstruymcm.'11lismust bebalancedwilhu»racr
nuu
celluloserequiresnnaggressive hytlmlysis.Traditionally,Ihemonosaccharideswcresep:lr.ued hypaper~li mlllatugrallhy((IJ I. '111e individualmonosaccharides could then bequmllificd using.ror example,Ihe Nclslll1-SullHlgyi cotonmcutcassayfor reducingsugars (67).Ho wever,mcihodsbnscdonhighpcrtonunnccliquid chromatography(HPlC)orgascllromatngraphy(Gelarc nnwdUlIlhmnl.
Themost successful HPlCmethods for monosaccharide separationuschll)-c~ch al1gcI\'sills.
However,inmany casessizeexclusion istheprincipal scpamorymechanism.I'mcxumplc. Paicc ct at (66) useda HPX,1l5lead len exchllllgecolumnwithdciuniscdwater aslIIuhilephOlSC.'1111:
disaccharide cellobiosewasthe first unulyteto be eluted.Morerecenll)',a rncllw)(J'llrIIIC separationof woodhydmlyzatcsbasedon trueion exchangeprincfplcswaspuhllsbcd((Ill).
-IS-
Because~haridc:sdonot conUinchromophores or fluorophoru.refractive index is the most commonlyused dell:clOfinIiPl..c.110000'C:ver.IIhas poorsensitivity(0.1 pg/pJ) detectionlimit).
Byromparison.pul\CdamperomeUicdetectionisquite sensitive(0.1ng/JIl)deu::aioolimil)andwas usedin!he inveSligalion of thewoodhydrolysa\eSby Edwards <:1 al. (68). AII<:m3livc:ly. the S:K:dwid~can be derivati1.c:d10increa.'iC!heirUV Ofnuc rescense ccnsitivily.
CH,'"
HC-OHfHOHO~"O" =
HO- fHHC-OHIO.(Jlucopyrnnosc I
" 1-
0"HtC-OH
! ..." .
-r-
HC-Ok HlHe-f-OOORI ACCliCAnhydridc I
kO-CH HO-CH
I I
HC-Oo\c HC- OH
I Pyridine I
HC-I QAc (orl·methylimidazolel
"1- 0 "
HtC-CAe Ht C-OH
GlldlD!.Hen ttWll GlucilOl
"'il!ur e 1.6:,\ld ilnlAcelall: D.:rivlllil.atitm. Conversion ofD-GlucopyranoseImo Glucltol Ik~ xeIJle.
-16-
Mono-andoligosaccharidesarctnvotarttc anoncrerorc lllusll lCtkrivOltilCtiifa GC lllcth\1I1 is adopted. Atdilolacetatederivitizalion islh eproc edure most widely used antiisillllslral~1illFi~.
1.6.ThoughUlis method is relativelylaborious. only oneaccunepnlduc t ispmdurcdpcrs;leeharitic species.Thisisrotthecasewithotherproceduressuchaslrimelhylsilyat hlll.Inthei r11iseussiuIl llf atdrrotacetatepreparationandanalysis.Fo xcrai.(69)review edthecolumnsamlstali un:lry Ilh:LscS used10 scpar..ate aldilolacetatecompounds. Cnp illarycolumns oncrsupc riurrcStll 111itlll:1lIl!pcnk shapewithrcsrccuopackedcolumns.Inaudition.themorepolarthestathmaryphuscthereneernc resolutionbetwee nalditolacetateswithinucsameclass(l.c.•pcll1USCS.hC M~S .etc.},'Illetlame ion b:ationdetec torismost commontyusedand is scnsntvccnougnfurII1Uslpurpuscs.However, sam pleswtur acomplexrnarnxmay requiretheusc IIfamore sctccrtvcucector.In thesecaSl:S selectiveion mon ilori ng withalIIass spcctmlllc te rWtlull1 hethe :IP]1lllpri:lledete ctor.
1.4. Pyrolysisof C:lrboh ydrat cs
From ananalyticalpofntofvie w.py rolys is orrcrsanalternative mcthtldtlfcha ractcri,.i I1~
thesefrequenllyinlrJclible polymers.uowcvcr.uerchas alsobeena greatneatIlfinterestinOlPlllieci py rol ysis forthe productionor use fulchemica l.~fromc,ublll1ydrale·cunlaini llg biomass(7m.
1.4.1.Carbohydra te PYrlllysisMcchallism.~
Thepyrolysisofpolysaccharides.and evensimplemonnsacclrnrldcs,C,LOpnduce ,I wide varietyof produc ts andunravellin g aume potentialreaction mechnnlsmshas pnwedIMI simp lel'ISk.
Forthe sake ofslrnpllchy and becauseofthewidesp read Interest inil.~pr npcr ucs.ccuutoscli:l~been thefocu s ofancnuonformost researchers.
Shall zadch dividedthe pyrol ysisof celluloseIntothreedistinc tive. tempe rature related pathways(71).Low temperature pyrolysis(ambientto30lY'C)slow lyyields char.WOller,CO2and CO.rmcrmcnnetemperaturepyrolysis (300-6O(J'C}prod uccs mainly1.6'arlhydro·B.D·glu copyrnnosc amiother relatedcom pounds. Above6<XfCgasificationof thepolymcr occurs.Inmanyinstances all threep"K:e!;.<;<:smayoccurduringthecourse of pyrolysis.Other parameters,such as the time- tcmpcnaurcprofile,thepresenceof inorganicimpuritics ,and therateof removalof thepyrolyzatcs from thehealedzone,canhaveallignin eant effectonthenatureanddistributi o n of products.
TIle principleproductoffastpyrolysisis 1,6.anhyd ro·B-D-glu copyr:mose (levoglucosan),I.
111llUghthetruereactionmechanismhasnotbeendetermined,a number of thoories havebeen pl11pnscd.Gotova suggestC1J thaipymly ~isisinilialedbyhomol ytic fiS!.!onOIlglycos idicbondsites dispersed thrllushoulthe cellulose molecule(72 ).The polymerIsthenrapidl yunzipped viaaself- pm pnglllingradlcal rearrangementto
rorm
levogtucosan(Fig.1.7).M\I.~tresearchersdtscouraunsin favourofaheterolyti cfissionmechanism . Essiget31.(73) Ilmpuscdthat hreak ageuf theglycos idicbond producesareson ance stabilised carbocation which then undergoe san illlr.lllluiceular addition(Fig.1.8).Thc process ispropagatedby theccnunuarlossof thelcvUl;!UcIIS;\ll endullil.
However,pyrolysisor cellulosealsoproduces 1.6-anhydro-B-D-gtuc ofuran osc .II,at approxlmutelyI()% thc yieldoricvogi ucosan,andncithc r oftheabov emechanisms can accountfor ilsrornuuon. Sh.1fi....adellctal.(74)haveproposedthe gene rationor a number oflntermc dtatc :lllhydmsug;lr svialhe nuclcnplnlic displacement oftheglycos idicbond by oneofthe ring hydro xyl gmulls (Fig. 1.9).Oneofthese,1,4-anhydro- ·D .gluc opyran ose,
m,
could then rearrange tofonn·Ill·
(In
(III)
(IV)
oo HO~ ~ HO~~R +1;;j;p'
RCpcal oI(ll I) :uldrlV)
Figure1.7: HomolytiCFissionofCellulose(72 ).
an
·19·
JL:1
oJ«)~o~,. ~;" ~·
R"
'"(- ;1;j;/ oo~ ~'
Ioi gurt1.8:IlclcmlYlicNu ionolCcllulosc(73).
.z..
,,",~ fQ '''' ~H '~
v::-v ,,,,,,O OM a OH
H,OHoo~
,
I
O ft
OH In-2 CELLULOSE
/
~,,,,
~,
~OH
(III)I.4-Anhydroglucopynnosc 1.!·Anh~lucopynnoK
-.
OO€1 ~OH
'"
(TI)l.6-AnhydroglucofUl11nOSC
av>
1.4-3.6-Dianhydroglucopynnose Fi~rl.9: Pyrolytic P:1thwayfortheFoml:dionof 1.6-Anhydruglucofuroull!'lCf741.either levoglueosm,I.orl.6-anhydm·lj· D-glucofur::lnosc.II.
A large numberofsnancr.!1Ofl·.\'PCcific~'()mpound.'Clll1belimnedduringpyml ~i:'\.Indeed.
Pouwels et:1l.pyrol)udmicrocrystallinecellulosebyeuric-poinl-GC·MS:uJddetectedWIpyml)'7.alCS
·11-
(75). ManyofIhe larserpymtyzatcs, suchll~ lA::l.6-!lia nhydm-·D.glul11pyr.II1\I....••IV,~.
hydroxymclhy!.2.furaldcl1yde,V,aOll2·fur:lldcllydc,VI,are cllllr.lclcristic pnll\ul1SIIf cellulose pyrolysis and Shafi7.adeh(71) has proposedtlmtme seproductsare funnedhythedcgrad;uiun Ill' levoglucosanandIhetmcrmcdtarcanhydrosugafS(Fig.1.10).TIlesmallerllnltluclslireliJnlll,'d hy avarietyof multistep pathwaysinvolvingIheruptureof the pyranosering.The y1lI:1)'Ilrigil~llcfrum lhcPYra.MSCunitsinlhcpolymerchainor from till::lUlhydnlsugllrs,lLml:Inumherofmcchnubaus havebeen proposed(75.76),
1.4,:Z.Malri xln nuenCC!I;
In addilion lo tilOsepamrnelers discusscd inscc liunl. I.3. lheCIIIllI1l.1Sitiunl lf lhcm :llliKC;\l1 have aprofound tnnucnccon thenatureofthepy rol )'sj.~pnutccts.Fur example.ilh:l.~been.~hllwn that aclds cnlmnccIheronnauon oflcvoglucosenolle fmm ccuoloscn the eKpcnseIll"ICVllglucClslLlI (77).
Thepresenceof inorganic ash is cspccranyhnponantInthepymlysisufellrIKlhyll l'llle.~.f-or example,Essig1.'1al,found thalaslittleas0.05%NaCI(w/w)elmrcdoccIcvllglucnsllllyichlsInnn 55%10 9%(78).There wasa corresponding increaseintheyieldsofclmrandUghlV()I:llilc.~.
1.4.3. The Specificityor Anhydrosugars
Anhydrosugars areuconly ehromlllogrJphahle pymlyza tesIhal retainlhe origin;11 stcrcoconnguraucnof lheparentsaccharide. Budgell 1.'1at.(7!JJdCJllHl1.~lm!ctlIhal unique anhydrosugarscouldbeproducedfora variety ulhcscscxandpcwascs,andthe ycuuld heresolved
-21-
(I)Lcvoglucosan
D·Glucosc
I
/ ! ' "
-Q o
Lcvoglucosenone (IV)I,4:3.6-Dlanhydroglucopyranose
He=0
~=<)
t~
I
H io H
HIOH CH~OH 3-Deolly-O-eryllV'(l-hellOSulosc
(V)S -Hydrollymelhyl-2furaldcbydc
.A...?-CHO H
(VI)2-FuraJdehyde
Fi~u re1.10:Ponnauon01'Non-Specifi c Carbohydr:uc Pyrolyzates(71).
. ".
crsor
mq ID~
Q"o"
011110
I
011O-GlucapynnCM'! J,6. AnhydroglutopytUlOsc 1,6·Anhydroglucofuranose
01fJI ,.---()
QIl.OII I D g
110,
D·Mannop)TlIIlOSC 1.6·Anhydrom ~yrU105C
IDei
OfI
ri:'?
U
011C.1JPlI
1~II'OI[
Of
~\lll.Oll
11O~ , tll
D·Xylop)T1lOOSC
cnon
~ll.OIl
al L·Arabinollll'll'lOSe
1,60AnhydrogaiaclOp)'rInose 1,6·AMydrogalIlCIOfunnosc
~
0111,4-Anhym-oJO;ylopyunose
~
011 JA·Anhydroanblnopyranoseri~un'1.11:111<:SaccharidesFoundin Wood<lnd TIleAnhydrosugnrsTheyFormUpon
-24-
using polar phase cnplllary gas chromatography.TIlis sumc systemwnsusedIII maractcrlzctlre saccharidecomposnlonof a number ofhomo-and hetempulysaccharidcs(KO.4Jl.InICf\)stillgly.
electronimpactmassspectrometrycoulu uistinguish betweendifferentclassesIll'allh}'lImsugar.;[i.e.•
1.6·anhydropyrnnoS(:s and 1.6-nnhydrofumnoscs)but notbetween ,mhydnlSugars within the same class(r.e.,1,6-anhydroglucopyranoscand 1,6-armydmmannopymnosc).11lCsOlceh:uidcscncuuntcml In this s! udyandtheanhydrosugarntheyproducewhenpy mly zcd arci1IuSlr:HcdinFigure1.11.
1.5.Objeclivcso fnisStudy
1bcobjectiveofthis workW:lS 10dctcr mtncttcuscfullness01'
oo-unc
l'y·GC(·MS)us a qua inanvcandqueunanvc toolrorurcinvesliga1iun ofl heS:lccharidc cOlUlKISilililiufwCMlllllllllls.'rradtttonatmethodsofsaccharidenn atystsmehlllgandarduousami Ilunultcuducmsctvcs III routineusc.Bycomparison.Py-GC ortcrs uc mivanlllgesof specd andsimlllidlY.Moreover,ithil~
beenshown 11m! uniquepyrnlyzalc~canheproducedfurncutmls'lccll:1ridc.~(7').
Prior10thisinvcsliga1ion. anumber ofpyroly/.crswerelnvcsugmcd;l~Intheirsuilahililyfur carbohydrate annlysfs.The Instrumentswerejudgedoncrhcrtasuch:L~caseofsampleweighingand loading. reproducibilityofpyrolysis. and crrccuvcrc ssofllymlYl.atetr ansfer. Wherepmsihlc. hardwaremodifications weremade10 meettheabove demands.Tllo.<;(:pymIYI.cn;tll,llwereI'cllliid tobemost suitablearc describedin section2.5.
Initial experimentsfocussedon thepyrolysis of isolaledpolysacch:lrides111:!1were representativeofthe ctrrcrcmrarbonydratcclassesfoundinwood.111e tnnucnccof parameters.such as.hepyrolysistemperature,inorganic ash and
ns
removal,andinn exchange.were illvesligMcd.-25-
Neill.,lhcsacd1aridccomP05jtion.~ofa numberofwoodpulpswerecharxtcrizcdbyPy.QC.
Thepulf15cho sen werebothhardwoodsamisoftwoodsandweremanufaauredby •numberof differentrecccsscs,Where necessary.pymlyx..alCl werek1cntificdbyPy·GC MSusing boIh electron lmp act(EI) andchemicalionil..1lion(el).Hereagain.uemnuenceof a number ofparameters such 3SmClhnd ~ofash removal, Ion-exchangeandscspcnslon pitwereinvestigated.Basedonthese results ,llfllimb: etl pre-iream cruproceduresfor pulps of diffe ring chemicalcomposition were dcvclnpctl.
Finally,wylical~swereevolvedwhcrcbylhesacdwidc eomflOSilionorthcpulps wasqU;ullinedbyPy.Qc.O:xup;1riSOOSweremade:with results oblainedbyclassicalmethods or amlysi5.
CIIAPTER 2:EXPERIMJ.:NTAL
2.1.Material s 2.1.1. Chemicals
Triflu o rnacctlc acidwaspurchasedrromSigmaOlCmicals(SI.Loui s,Mo.) . Allal:lrgr.ul~
hydrochloric,sulfuric.nil ricand acetic acids. amisodiumsulfitewereparcbascdrnu» BUll(Tu nllltll Ont.). CertifiedACSgradepyridine.mcihanol,chlom roml.dichlofllllle Ul' l1"le.:l1ld1elmhYllru l"ur.m werepurchasedfromFish cr Scient ific(HaUfa,... N.S.). Sodiumberuhyrln dc.accticanhydride.and l-methy hmldazolcwerepurchasedfrom Aldrich (Milwaukce.WI.),Allmeutch luridc.~amlthc Ambcrlitc'\on-exch:mgcresin(WramI)werepurchas ed rmmCanlab(Mi ....~iss,llIga.Onl.).
2.1.Z. Carbohyd ralesand Wood Pulps
Amylose.
«ccnurose,
locust bcangum,amhinog alac..1an. xylanfnlllluatspclts.1,(IOulhyd m·(l·D·glucopymnose.andall monosaccharides were perchuscdrromSigmaChcmlcrls{51.tuuis.Mu.J.
Standard 'IIdilolacetate mixtures werepurchased fmm Pierce Chcmhals(H I)C~llllll. 11.).
Mlcrocrysiadinc cellulose (I1.Cgradc) was oblaiflCdfrom J.T.Bakcr ChcmlcalCIlIllP.lllY(Nl.).
Avlcelwas obtainedrrom FMC corroranon(PhiladclphJa.I'A).Theanhydmsugarsl'lIl1lard~ .1.<1·
Anhyd roxyln py ranosc. J,4.anhydr oarabin opyrano sc, l.tl-an hydrllll1'1I1Jlll/lyrallusc. Ik anhydroga laclopynlllosc.anti1,6·anhydrogluco furJnuscweredonatedbyDr. A.S.Perlin.Mt:Gili Univers ity.
The blackspruce:andbirch rncchanlcatpulpsandsprucekralt/lul p~wereagi nI'IlN1JDr.I'.
Whiting.Abitibi·Price.Onl.TheMacksprucesuuue pulps werednnaledby Mr.Gu rdunIlnwlc rick,
-27-
FUJeifTcch nlllugiesLahc~IOf)'.NorandaTccIvlologi esO:nm.f'Q.TIlt:aspensteam-a.plosionpulp wasuhtaincdfromDr.B.V.Kotll, CRPP.Univenilfdu Quebec 1TroisRivi~res.Thespruce Ihcml.1lmcchan ialpulpandlhc5p1ICCmilledv.ooodIi".inwere a gift fromDr.M.jceen,Swedish PulpandPaperIIt~itule.Stockholm.lbcbirthorganosolv pulpwu obtainedfromMr.J.Motsey, AleellCu.•Newca...uc,N.B.
2.2.")'drul~slAldiM AalalfDerinlizat ion preeedures 2.2.1.bllialcdl'uI Y~llccllUidcs
Polys ac charldcswereh)'cJroIY7.cdinamannersimilar10111mdescribedbyHondaC(til.(RI).
Samplc.\(S-IOlng)weresu.'iflCndcdIn 2 M mnuoroaceucadd(TFA)InISmLmuMtoucncd flasks.TIICna.Jo:swen:stnpperedencrpurgingwithnitrogenforafewmimncs.Thiswas doneIn order10preve ntdcgr.:td:llionofUtemonosacdwidcsby oxygcn duringhydrotysis.lhc sam p leswen:
IhenplacedinaIOO"Coven for six hours.AOcrcooling.thes:l/!Iplcs werecvapor.llcdto dl)'TlCSS un anll ary cvaroea or.It.~allaliquotormeth anol(1:roL)wasaddedtocadtflaskand the h)'c1nd)'1.alcswerecvaroratcd10dryness once~ain.Thisproctdun: was repeatedunlit theodour of TFIt.cou l d 1"0longerbedt::tcclcd(usua llythree limes).
TIle1Il11l.~ccha ridcswere;ma1)'1.cdastheiroorrcspondingllIditol acetates .Thehydrolyz:llCS were tlissulvedillwaterOmL) anti 1-2tugorNaBH.(:Ired uc ingagenl)was added10cachflnsk.
Thesampleswere allowed10 s131ll1atroomicmpcraurc for(Inchour.alwhich limethe excess hORlhYllrhJcw;~~l)Culr.tliscdbylhceutuonorafew dropsof,50$acetic acid.Thesolutionswere cvaporaedrod()'l~ sSand lheborate wasremo vedrromIhc113J.llplcSbyevaporatingihrec timeswIth sma ll31iquolstlr4:1lllClhOlOO11acelicacid.
-28_
Aceticanhydride(I mL) anddry pyridine(I mL) were added 10 lhcnasls,whichweremen stopperedand Incubatedat 10000Cfor onehour,Attcr Iheaddilionof waternUlL)thedcrivmil'l'l!
sotaucnswereevaporate d \0dryness.Bvaponntonwas repealedthree rimeslinerthemhliliunof small quantities ofmethanol(2-3ml).Thcdcrivatizcdhydn llF.aleswere dissolvc.lin chl\lRlfllOlI (lmL)andtheinsolublematerialwasremovedby passagelhrough IIgla.~swoolrutcr.111Cmurkl bottomed flas k wasrinsed with O,SOIL ofchloroformwhichwasalsolillerell andaddedhilhe original solution,Thecmcrorcrmwas removedunder a slream of nilnlgcn,U\llthedriedt1erivaliv es werestoredina Irccz c r unlilanulysis,
2.2.2, Wood Plllps
TIlehydrolysisprocedurefollowcdwasthmdescribedbyFengeland Wegencr furcellulose- containin gmaterialwhh a highlignin cement (fI5).Pulpsampleswere cxtractcnrorrUUT 10sixhours wilhaneUlanol/hCnzenc(1:1)mtxnrrctc removelipids,waxes. ere. TIledried puIIlS(15-)O1Il~)were suspended ovcmlglu in 5gofanhydrousTFA. Aner refluxlngror onehour,
me
lLcidwasdihllctllll 80%withdcionlscdwaterandrcfluxedasecond umcfor twenty mtnurcs.,11eSl1lnflle,~we rerun her diluted 1030% and rcrluxcdagain forone\lOUT.Afte rfillrJlitln Ihnlughgl a~,~woolthe sululilltlS were cveporated10dryness on a mtaryevaporator, TIleresidualTPAWilSrcnmvc d hy rcpcalcli evaporation wnn methanolunlill hchyd m lyzaleswereorour free.Allose.Iheinternalstandard,wasaccuratelyweighedIntoeachna.~k{2-}mgj,arll,lme sam ples warsdissolvedinwater (31lI1).Thehydmtyzutcswere red ucedby Uleaddition (If.'lIKlium borohydride (5-6mg).Aftcr one hourthe residualbomhydride wasncutrauscdbyU,eat/diliouof afewdrops of 50%aqueousacetic acid. Thesamples were theneVilf/llraledto dryne ss, 'rhc
·1!'·
';C(:lylatillllproccdureIsba.~edon thatdescribedbyBlakency cr aI.(82).'mereduced saccharides were acctyletcd atroomtemperaturewith acetic anhyd ride(2ml)usingI-methyllmtdaznle(0.2ml) ascatal)');!. After10minutes,water (S ml) was added 10 each flaskand !hesampleswereallowed tosit unlilcrxil,Thesampleswereltansferrcdto2Sml screwcaptesttubes,thereactionflaskswere rinsed twice withsmall aliquots ofwater,andtherinseswereadded10theoriginalsolutio ns.The solutinmwerecxrrcctcu threetimeswithdlchle romethanc(2ml).Thedjchlommcthnnc enra c tswere pouled,evapuratediodryncssunder nitrogen,anduo rcdlnaIrcczcrumtlanalysis.
1.3.GC Analy.sisllfVel"i vativcs
TIl(:aldilOlacetatederivativesoftheIsolaicd polysaccha ridchyd rulyzalCSwcreesso rvcuin 0,511I1of chloroformandanalyzedusingaVarian 3700gaschromatograph (Georgetown, ONT.) equipped withJ&WDB·225 capillarycolumn(30 ml(0.225mm,0.25urn film thick ness, C1lRJl1Hltngm['lhicSpecialties' l)rockvi11 e. ONT.)andaflamefcnls auondetector.Theinjectionport and detector were maint,Lined nt 250"Candtheovcn tem peratureprogr;lffiwas as follows:200 °C for 2min.,3"Cmin·1until235"C,holdfor15min.The column flow wasI mllntin.ofheliumwitha splitrauoof 1:30. Theindividualaldilol acetates wereidenlifiedbycompatingtheir retentionlimes with rtmscnfcollllllcrcially availablestandards.
TIw::nldltolaceta tedcdvntlvcsisolatedfromthepulp samples,however,gaveunaccep table ctmnuatngrarns on neDB·22."iphase column,most probably dac todcrcrtorcncnincol umn pcrrllnll;UII.'C.These~,mlples werescpuratcdinsteadon a CPSIL-19 column (30III x0.25rom, 1.2 1l11lmtckncss.ChmmpnckCanada,Blenheim,Ont.)undersimilar conditio ns.QuanlificlIIlon of the individual saccha ride
ucnvanvcs
inUtepulpsampleswas malic by compa ringtheir peakareas with.JO-
thatof[heIme rna!srancaru allos e .Therelative detecto rrespon sefor eachtnunosaccarulederivative withrespecttothator theinternalstanda rd. allasltol accrue.was deter minedprcvinu.~ly.
2.4.Sample Prepar ationforPyr(Jlysi.~
2.4.1.ProlonExchan geaf lsolated Polys accharide s
Becauseof the diversenature ofthecarboh ydl'illesunde r invcslig :llhll1not11~mctho\lfllr metalionremovalsuiledall. wnrer solub lerolysilccharidcs weretrcmc dusingilsll1'l1~
canon
exchan geresin(Ambcrlilc H'Innu), An 0.5%w/vsolutionwasusunttyprepared ,theresinWilS addedinaratioor 10:Iandsllrrc<ltilr4nours.111C resin W,lstllclIaUllwc dIIIsettleuerlthcIfllllll~l solulionwasdecanted.The polysacc harideswererecoveredrromsnluuon euncr hyIYllphilisillilUI or.ifpossible.byprecipitalion
wnn
anon-sctvcasuchiL~tClrahydmfu r.U1.Inlhe innerpRK:eSS, finelydivided prodUCIwasIlhlalnc tlwhenthepolysaccharidesotunons weremilled.~I(lwl yIIIat least twice tile volumeof ion-a uvcmwhilevigorously agHiltln gIhemixture with a high ShC~f horuogcetsc r.TIle polysaccharidesWCItthe ncullcctcdby Iihratton,alr·d riedlure mov el1Iuslufillc volatilesolventandfinal lywcn-drlcdal 75"C {[lr eightlimITS.wacr-t nseiubtccarbollydr alc5suchas cellulose werepmtun-c xchungcdrullslellcctivclyhy suspending0,5gof samplein100rutof 0.1NlIelfor 4 ho urs.TIle s:llllpleW,L"mainlaiucdill suspensionbygcr ujestirr ingwithamagnelic sttrbar.Amyloscwaspmhlll-exc!limgcdill lhismunncr u~i ng0.1NHelil150%aqacousethanol, The solidswere thenculleCledhy liItmlio n,washcdwilh 200mlof deloniscdwater(50%aqueous ethano lforamyl(l!'C)anduvea-driedat75"Cfor eighl !lllUffl.
-3 1·
2.4.2. Pre tent:xcha nllfof WoodPulps
Aft.cr Slime initialpyrolysisinvestigations.themost suitablemethodwasfound 10bethe following. Pulps were delontsed by suspendingO.5gof thedriedmaterial in100 mlof0.1 NHO fur 4hou rs.nl\~ salllple.~werethencoltecled by filtr:llion. washed willldeionlscdwater (200 ml) to relllllveany residualacld and oven-dried for 8 hours atSO"C.
2.4~1.pI(Adju.~menl llrAcid-WashedWood Pulps
Forthe studyorthe rnrlucnccof suspension pH on\Ile anhydmsugaryieldthe procedure dc.<;crihe()inIhe1a.~LsccnenwasmudifiedLa includeanotherstep.Anelacid-washing andfiltration, Ihe Mlll fllcswereresuspendedforISminutesinasolutionwhosepHwa.~adjustcdusing0.1N HQ andNII~OHsolutions.Thesampleswere collectedbyIiltratlon, washed witha very smallquanti ty ufwarcr(20 m!).andoven-dried atScrC. TIlefinal acldityoftheIsolatedsamples was nOLmeasured.
2.4.4. Carlon)<;XcllllllgCofSulfitePulp s
Sulfltc-trcntcdpulpswere nrsrsuspendedin0.1N Hel for2hours, collected byfiltration and washed witha sm:1l1 quanlilyof dclonisedwater (50 ml). Thepulps werethen suspendedfor2hours in a 0.1N sulotiunofthe calionunderlnvcsugatton.111echloridesails wereusedin allcases. The pulpswereIhen filtered. washed willidctontsed water (500
mn
and driedovernight at 5O"C..)2·
2.5.De scription of r,roly un
Initialexperimentsfocussedon develo ping3 reproducibleandea...yto uscon-linep~nllysis. GCmethod.Twol~pcsor pyroiyecrswerefoundtomost suttablc andtheyarcdcscrihcdtnne followingsectio ns. Athlrdp~ roI Y"lCr,theP,lCkanlModelt\91CuriePoinl Pynllyw,wasabu Investigated.Ho wever,whilesolublesamples couldbecoaledon the curie
pouu
wires.itpmv\~1 impossibleto satisfactorilyapplyinsolublesamples suc haswoodpalpfilm:s.Purthcrreurc,willi regard to reproducibililyandQnhydtu':iugar yield,Lhe resultingpyrngr;unswere inferiortilthm;e producedbymeonenwopyrulyzen,;.Because(Ifthesepreblcms,this]1yrol~I-erwas nutUSCtlinlhc followingInvestig mions.1.5.2. CDS ryroprube120
ThePyroprobc120(COCmicalDataSystenl~,Oxford.Pcn nsyl~;ulia)cUllsisls (If1Ipmhe1111\~1 wilhaplalinullIcoil(Fig.2.1).1lICsampleisplacedin a qUl1n1.tubefillctlwilh3pu n m.sllu:ll1zplllg whichislhenInsencdintoth ecentreoflllecoil.1l1Cco tI is heatedresi.\1ively toaIlrcsetlclllper;tlme andthepymtysetc sarcswe ptbythecarrierga... flUllllhctubetntolhelucrruccoven:UKI fin:dlyinlu theGC injecllonpori(gasnowrae==13.7mU ntin.),The Interfaceuvcnis mainlained:112.'i(J'C In order10prevcrucondensatio n
o r
mepyrulysalC.1l1Cintl'rfacc designwaxmndilicd !itllhat tile ernuer afromthe pyroJyserpassedthroU&!1a22gU3gene e dlebeforeell!eringIhelnjcctkmpon(Fig.2.1).Previously.the pyrotyscrwasattachedtulhe GCinj ecliunport bya 5 emlung.slainlesssteel, wide-boretube,Thereproducibililybetweenrunswaspo o r,mllSI prulJalJlydueIuhrcruupletcIIII"illg cruepy rolysalewithjhcearner gas.Also.bcC3USCthetullCwa.. unhealed,merew:l"lhe stnlllg possibtlttyorpyrolyzale ccndcnsatlononlhe internalsu rf acesnl"Ihetube. Bycomparison.the
·33-
ProbeFiltedwith .PI.tinwnCail
\
GCO<rnWill
Ftgure2.1:CDSPymprobeJ2lt'.
l11utlilicoJdesignhas 3 verysmalldead volumeanti isshorter(1.5emin length).Repeatedpyrolysis ur isolalctlpolysaccharides. suchas
nucrccostamre
cellulose.wasroundrc giveanhydrosugaryields lhalvary hy 2%ortess.TIlesan pfcs.USUally100-150)Jg.were weighedinto the quartz tubeusing a Perkin Elmer
Autobalancc AD·2Z.ThelUbe W,1S filll:dwitha smallquan tilyof quant.wool inunlcrtu f,lCill!alc sample handling. Unlessstaled cibcrwlscthes:lmpleswerepyrolyzed :It5IXt'Cfllr 211 seconds:Ukl notemperaturerampwasused(i.e.,maximumhealingrate},
2.5.3.SGEPyrojector·
The Pyrojeeto r(Scientin e GlassEngineering,Auslin.Tex:L~)consists of a elllliin\lously hC,llc\t furnace linedwithaquartz tube(Fig.2.2).Samplesareinlruduced intu rncpymlyzcr thnlul:h:I normalGC injectionportusing asolids inJeclor.The septum ispn::·dri llctl tu :IJlOWeusypass:lge ur theinjecto rbarrel. TIlepyrolyl.:Ue.!iare swept intoUleGCinjecliun pUrl hy Ihecarrierg:l.~lhnll.ll:h aneedleinterface sim ilar10uuudcscnbcd intheprcviuus scclillll(ie .•luIalt:a."nnw mlC"13.7 mL/min.).
Forquersnanvcanalysisilisnecessary 10 ucicr mmctheweighloflhe sample bc rore py rolysis, and uus proved difficult 10dousing a solidsinjector.Alic rsome!:lVes lig:llirlll, lhelIIusl suitibleweighing proced urewas foundto be thefollowing:The injeCltlrwaxdisas.~clIlhlc(l,and rue injectorbarrel was weighedon the AutobalanceAD·2ZThesample (J(){)-15nIlg)W:L'I lmroduccdIntothe barrel ,which wasmenreweighed.Arterrc'l~SClll h lylhe s,ull[l1cW;l~injecred lulu thepymlyzcr,TIle py rolysis temperaturewas SOirc.unless stetcdorhcrwlsc.
The solidsinjectorsuppliedwiththeryrojcc lorWiL~fo und10hesuitahlefur meinll'llduclilJn ofmost Isolated polysacc haridesInto the pymlyzcr. uowcvcr, theintcrnnldiameter Ill'tile injeclllr barrelwas 100smalltoaccomodaelarge, fibroussamples suchus wllodpuips. ttcrcrorc. 0111h\jccwr withalarger bore barrel(2 mm. internaldi,ll11ete r)was fabricated.The1x'!ClhtllughjhcinjcctilUl
H.~ ==ll=='"
GCOYenW.n
Fil!url!2.2:SGEP,'mj ccl orr.
-J5-
~==t==_ SeptumPurge MicrofllJTll,l:e Oven
QuaruWoolPluS
.J(j .
head assemblyoftilepyrol yscrwaswidened10 facilil:lll:,hc larger Injl'\:ll\f.l11cnew iLUl'ClllTWlllk,'I' wellformepulpsamplesand theminormodificalions did netarrccr lhepCrrUnl11111CCtil"11k' pyrotyzer.
2.6.Pyrolysis .GasChromatograph y
Each of thepyrolyzcrsdiscussedabovewa.~nttacncdIII thelnjccliullpUll of,IVllriml:nnu Gas Chmmatogmphequippedwilh11 namelonisntlundetectorand nJ&W [)1I·17UIrusedstucn capillarycolumn(!pmttrlckress,30Mx 0.329111111. Ollumilltlgr.lpl1icSpcdnUies,Ilmckvllle, ONT.). Poorresolution of lheanhydmsugnrproductswasIlhlaincdifth isL~llumu(ITlise~llIiv ;I1cl\l wasnotused, TIle illjeelionPOlland detecto rwerem<lilll;lil\cdIII 27l1'C.Thc tcmpcrnmrcJlTugrilill SClling.~were asfollows:1000C.hold for 2 minutes,increase 5''Cmilf 'un!i1 2MY'C.hahlInr5 minutes. The spIllwasmeasured al10.5:1 wilha colunmnow rate 1I1' 1.2mhniu'. The chromatogramswere acq u ired usin g aSrccll",j·l'hysicsSt>4290inlcgr:llurlinked.viaaL;lhnct lntcrfacc.with aTandy1200liD personal compeer.andcUnlmllcdbyIISpcClra-PhysicsWinllcr"
data system (SanJose, California).
2.7.Pyrulysis .GasChrolllalogr~ phy•Ma s.~Spct tru lIlt lry 2.7.1.EleclronJrnpaelloniz3lilln
Pyrolysis-GasOlromnlography.MassSpectrometryusingctccmmimpattiuni"l illiul1.I'y' (; (;' MS(EI) , wascarrlcdoutbyaltm:hingthc Pyroj ttlor LO me injetlinnronuf aIlcwlcll f'ackmdModel S790AGC/MSDequippedwlthaHP S970AWorkslatiun.TheGCW;\SlincIIwilha nunll;\!tore J&W DB·1701column(O.2S5mm x30 m.U,SJIm1I1ickncss).Many orIlic pyrllgrall1 s , c~pccialJ y
-31-
thewondpulpc';.CUlIlaio Ihinyormore peaks.1bcincreased resolutionobl:ailclby using a narrower boreculumnr.:aci l i r all~due idcnlilication of these peaks.1ltc:injca ionportxIdMS Inletline were Illainl:lincd at 27fTC.Theoven lempctilturcprogr.amand splil llowr.uewell:asdesaibcdinsection 2.6 .11Jecolumnnowrare wasImUmin.1bcc:1ectronimpact kJnIzationvoltag ewas70 eV.The m:l'i'iispcclromClCrwasscannedfrom30to250 a.m.u.
2.7.2.Chenlicallonizatinn
nle rymjcclor wasinterfaced whhavartan3700GCmeedwitha norma!boreJ&W DB_
1711I (O.25:irumx30 111, U.5umlhickncss).Theend ofthecapillarycolumnwas introducedInlo lhe1,....llllhin;LlionEVCISIIUn:c of a VO 7070HSdoublefocussingmass spectrometerequippedwith itOS2[)J5datasy!ilclIl .TIle chromalogrnphicconditio nswereIdcnlicallothosedescribedinsecllon 2.7.Ammoniaw...~used lL\\hi::rcl1gClllgasandtheionsourcepressurewu6xIt)"'mbar.The ieupcraturesuflhccolumn Inletandionsou rce were27O"Cand2OO'Crespcctivdy.Thesourcewas u-<;cdinlhe CI lIwdcandthek1flI1.OIl loo vollage was100eV.Tbemass spectrometerwas~
fnml6llkl2.'Wla.m .u.011Isperdecade.
2.8.IUOfll::micAnalJsis 2.X.1. Ashill~
TIle,Ishcomcmof lhe:polysaccharides andwoodpulpswas dcterrnlrxdusing(he ASTM procedureforashingwuou(83).Tbcs:unplc (1-2 g) wasplaced in a prcwclghcdporcclaincrucible:
3ndwas wcighcu III,hc ncarc51 0.1 mg.TIlecruciblewaslhcnplacedIn anoven OilIOS'Cforone hour, AOcrc(lI,lli ngin;1dc$ic;JI(l ffor30minutes(hecrucible+sample wasweighed.This
"'11.
procedure was repeated until UIC weightremained constant10within0.1mg. l11CIinalweight measurement isused10 detcrnnncd thetruedry weight ofthesample.Thecrucible W<lS then I'lill'l'ti onthelipofa600"Cmurr lcfurnacelindmecontentswereallow~'tItilignite slowly. AucrIII minutes. when lhemajorityofthecombusubic material nas beenremoved.the crueihlcisutnvcrltu thecentreof the furnace.whichis1111.'0 closed. ArteronehourIhc cructblcsareremoved,~111,1lcllin adesstcater,and weighed. Thisprocedure is repealed untiltheweightis COll.'ilant IIIwilhill (1.1IlIg.
Atevery transfe rstep the crucible is covered wilhalid inorder III preve nltheInssIll' sample.'J1IC ashcorucra isccrcnnrncd as a pcrccnmgc oflllCdry wcll:ht.
2.8.2.TheDeterrelnatlunof Irun inthe Acid-washed,Fcl·.ElIchall~cd,SlIlIil c· T rclll"dI·uljl.~
TIlIlaxhcdsamples fromtheFc;··CllchllOl:ed.sulfitepulpsweredissolvedIn2nu.(Ifullrapurc HNOJwithgentle healingifnCL'Css;lry. TIle soruuons were then dilutedIII 50ml.,The Intn ccnccrareuonwasdetermined using aPerkin-Elmer 231l0 :lllIlIliclItlsnrplitmspectrophnlumClcrwhich wasset tomonitorUleabsorption at24M nm.'Ihelnstnnncmwas calibrated with;1cununcrclal standardsolution(1-10 ppm).Sampleswlthnff-scalc absorbunccswere IlilutcdIllllunlil inr:llll~c .
2.8.3.SulfurContentAnalysis (l( theSulfite 1'tJlpsbyX.Ray Fluorescence
Thesulnte/sulfon ieacidcorucoror uicsulfiterUIp.~wasmeasured ind irecllyhytlclenllinillJ;
theirsulfurcomer susing an ARL8420+wavelengthdlspcrslvcX-raynuorcsccncc specrnnucrcr.
'Though methods do ellislfor determiningthereduciblesulfurcllnlenl in pulpsandpaper{1l4.IISl.tllCy arclaborious and sulfitecontainingsamplesarc not necessarilylJuantiialivclyreduced. lty compar isonX-rayfluorescencespectrometryisfastand accurate.atJdis mulilldyusedInqUilllllly
.39·
lhem.iljo relemenlillcomposiliono riI.widevarietyofsolidandliquidsamples.
Stand~!iWCfCprepared bymixinlfinely groundNiI,SO)withiI.rce-snmetrealedpulp«
0.1'1.sulfur).SiandardiIOOsamplepulpsweremilled10 lessthan60 meshwiUliI.TckmarA.IO aOilly(icalmillprior10 analysis.The:spectrometerwasequippedwithaRhanodeX-raytubesource opcr.lledal30kVand100rnA.andanargonflowproponionalcoemer cctector,ThesulfurK..IJ line:(26 :11O.68")signalWallmeasuredfor20seconds foreachsample. All measurementswere had,gnlUntlsublraclcd (20 :114.()(r).
CHAPTERJ: PYROLYSIS OF ISOLATEDrOLYSACCIlARIDES
3.1.Glucans
3.1.1.Microcrystlilline Cellulose
Microcryslalline cellulose(MC) isaverypure. partlculme ronu ofcel lulose.ItisiSlll:ltcd fromee-cclhncscafter intensive mechanicalandacid treatment (86).Ithas atowmolecular weight (i.e.•deg ree ofpolymerization,DP.,of 2(0 ). MefromtwodiITe rentsourceswere investig;ltl'd:
Avicel" PH-IOIfrom FMC and TLCgrade MCfromBaker. lIere, ns withlilt therulluwilll:
polysaccuaridcs.tilesampleswerepynuyzcdusing theCDSI'ywpm!lee.
Figure 3.l(a) is thepyrogralllobtainedbypyrolyzing AvicclMC ar(lIKt'C.Thepym!Yl.alcs were idenlified curer by comparisonofthcir retention rtmcs with\hllseIll'aulhenli!.:staml:mlsor hy theirEImassspectra.'rabtc 3.1liSlSonly theimportant carlxJhytlnatepymIY/.:lte.~.FurtherdisCllss;un on themcnuncattonofpymlyzatcscanhefoundinseclion4.4.TIleoprhnumpyrolysisICllljlcr;llurc forlcvogluecsanpmducuonwas found 10be between550 anti fiIKJ''C. Tile Avicel MC pyrogmm is remarkable for IlSsimplicity.especiallywhencompared with thalllhtainedIurthe samecellulosehy Pouwels etat.(76) whoused Curtc-PuintPy-GC ,ulalysis. Lcvogjucusan, 7.',dominatesIlle pyrogram and l.ti-anhydro-glucofunmusc,liS,is also present,tlMIUJ,lhin.~llJallcrqn.uuutcs. lIy comparingthedetector responsewithtbarof purelevoglucus:mstandard il hasbeend~tcnlli nccll h;lt 51.6%ofthepyranoseunltx inlhc ccucrosc samplewereL~1I1VCl1cllintnthetwu:lllhydmsug:u products (Table3.2).II was observedthat an oily.nun-volatilercsnfuccondensedatthe endul'the pyrolysislube anditspresence wouldaccount formuch ofIheremainingpYllllYl.llrc.Nochar remainedin thetube after pyrolysis.
