BASIEGAS; de‘TECHNIQUES IN ANALYTICAL CHEMISTRY SERIES BAKER + CAPILLARY ELECTROPHORESIS CUNNINGHAM +» INTRODUCTION TO BIOSENSORS LACOURSE + PULSED ELECTROCHEMICAL DETECTORS IN HPLC MCNAIR AND MILLER + BASIC GAS CHROMATOGRAPHY METCALF + APPLIED pH AND CONDUCTIVITY MEASUREMENTS MIRABELLA » MODERN TECHNIQUES IN APPLIED MOLECULAR SPECTROSCOPY SCHULMAN and SHARMA + INTRODUCTION TO MOLECULAR, LUMINESCENCE SPECTROSCOPY STEWART and EBEL. + CHEMICAL MEASUREMENTS IN BIOLOGICAL SYSTEMS TAYLOR » SUPERCRITICAL FLUID EXTRACTION Basic Gas Chromatography HAROLD M. MeNAIR, Ph.D. Virginia Posteshnie Tsite and State University JAMES M. MILLER, PRD. Drew University ® A Wiley aterscience Publication JOHN WILEY & SONS, INC. [New York + Chichester * Weinheim + Brisbane + Singapore * Toronto‘er GC pop epi by emi 024 Sent Pet Spon nthe nero Coe mau Taste pedo ie pape @ [At ah rene. ui imac hae {ht permite ty Sto? olathe 196 Unaed Seu Chr A Se pmon ft aye tte Cr min tte ‘asic gn cromtoraphy / Harold M. MeN Sues ies, a os SSS mes aS Contents Series Preface Preface 1 tntrodcton A Bric Hitoy Overview: Advantages and Disadvantages 2 Imirument Overview cases Gas Flow Control and Measurement Sample nts and Sampling Devies Colas “Temperature Zones Deters Data Stems 3 asi Concepts and Terms lintons, Terms, and SymborsThe Rate Theory A Redetition of H The Achievement of Separation 4 Stationary Phases Selecting & Colum ‘Castiation of Stationary Phases fr GLE Liga Stationary Phases (GLC) © Sold Stationary Phases (GSC) 5 Packed Columas and Ines Solid Suppers iid Stationary Phases Sod Stationary Phases (SC) Gai Anais Inet for Liu Samples and Solutions Special Columns parading for Capitay Colma (6 Capitary Columns “Types of OT Colunas OT Colum Tobing ‘Advantages of OT Colamns Colom Selection Capillary Inlet Systems Indes 7 Detectors Cawifcation of Detectors Detector Characteristics Flame foieaton Detector (FID) ‘Thermal Conductivity Detector (TCD) etron Capture Detector (ECD) (Omer Detectors 8 Qualitative and Quantitative Anais Ouattaive Anais ‘Quantitative Analysis conten si 2 s a S2SEeR foResaoe 1 1s 12 6 ug rs 6 16 an Contents 9 Programmed Tempertore Temperature Etfets Advantages and Disadvantages of PTGC Requirements of PIGC “Theory of PTGC Sper Topics 10 Special Topics oc-ms (Chiral Anais by GC Special Sampling Methods Derivatization 11 Troubleshooting GC Systems Appendizes 1. List of Symbols and Acronyms 1 Guidelines for Selecting Cepilary Colunas LIL GC: How to Avoid Problems 1. Calculation of Split Ratio for Spt injection on (OT Camas LV. Operating Conditions for Coplay Columns VE OV Ligud Phases Physical Property Data VIL Some Pressure Corection Factors ()) IIL List of Some Chromatographic Suply Houses TX. Other Resourses Aces of Applications Ades rn 14 46 ue 150 153 163 164 1 0 1s vo 18 191 1 3 4Series Preface “Ties ia the Techniques in Analytical Chemistry Seis address current Technics in general se by analal abortions. The series intends (0 Serve roa audizce ofboth prationers and cent of chem analysis “Tis audenceinclods not only aalicl chemists but abo professionals in other areas of chemi) and a otber dciplines eying 0 information Cervelo chemin analysis, Tu, the srs shoud be uct fo both Inoeatory and management personnel. ‘Writen for readers mith varying levels of edation and laboratory expertise tiles the series donot presome prior Knowledge ofthe subject, [nga the eae step by stop ough each tchngue- Numerous apt Cations and diagrams emphasize practi, applied approach to chem: “The specie objectives ofthe series ae + toprovide the reaer wth overviews of methods of naljsisthat inctue {bass inedcton to peniples But emphasie such practical aes {etechniue selection, ample preparation, measurement procedues, ‘ata analy quality contol a quay assurance: + to give the eeader a sense ofthe capbilties and linitatons ofeach techie. and fel fore appa to specie problems “to cover the wide rage of uf techniques, from mature ones 10 newer methods that are coming into common us: and + to communicate past loration in a readable, comprehensible Sle Reaer from he tecnica through the PAD sett o bora. Series Prece tory manager should come away with cate and coatdence about the {ee of the techniques Forticoming books inthe Techniques ix Anata Chemity Series wll over variety of techniques including chememetric methods, bcweso Sarface and interace analysis, measurements in bloga yes nde Sly coupled plasma-mass spectrometry. gas chromatopraphy -mas spe {eomcry, Four tansform infared spetoncopy, aad ther Sinica topics. The ers weleome your comments and sugestions tozaing atten ad future tiles and hope you find the erie stu rane A. Sern, Preface [A series of hooks om the Techniques n Analecal Chemis would be incomplete without a volume on gs chromatography (GC). undoubtedly the most widely used analytical technique. Over 0 yeas in development ‘Gc has become a mature method of analysis and one that Is thy 10 fade in pops. Tn the euly years of development of GC, many books were writen 10 lngom analysts ofthe latest developments. ew of them have been kept uptodate and few new anes have appeared, so tat satisfactory single inteoductry text dacs no exist. Ths book attempt 10 mest hat aed. Tt ‘bse in part onthe caler work bythe same ie, Basie Gas Chromaog- ‘rap, coauthored hy MeNairand Borell and published by Varia Instr ‘ments Some mater ako drawn from the eater Wiley book by Nil, (Chromatography: Coneps and Contras. ‘Weave atempied to write brie basi introduction to GC following the objectives fortes in this serie, It should sppeal to readers with ‘arying levels of education and emphasizes «praca, aplied approach to te subjec. Some background inches i required manly perl ‘tganic chemistry and some phy chemisry For ein forms cas ‘work, the Book should besiable for undergraduate anal chemisty Courses ad for intensive short courses of he type offered bythe America {Chemical Society and others. Analysts entering the Held should fad ‘ndapensable, and nds chemist working in GC should adit aut ‘elereace and guide.a Preface ‘Because the TUPAC ha recently published is nomenclature recommen ations for eheoatograpiy. we hive ted to use them comatenty (0 promot aunied set of detniions and symbols. Abo, we have endeavored {Dvnteinaschs way tht the book wouldhave the carasteisi ofa ile utor, asl especaly important for beinners in the el. Otherwise, he Content and coverage ae appropriately conventions "While open tabular (OT) sola ae the mot popular type both open ‘pula and packed columns ae tented throughout. ad the advantages, ‘tsodvantages, and apphieatonsarecomeasted, nado, pil chapter re devoted w each pe of column. Chpte 2 introduces the ise asta ‘mentation and Chapter 7 elaborates on detectors Other chapter cover Stationary phases (Chapter 4), qualitative and quantitative analy (Chop 'er8), programmed temperature (Chapee9), and woublesootng (Chapter 1), Chapter 10 riefy coves the important special topes af GC-MS, ile Ta + Oe z en where ds the distance betwen the peak maxi fortwo solutes, A and '. Figure 37 iustats the way la which resolution eaesnted. Taogens ‘ce dawn tothe inflection points i order to determine the with of the as at their base. Normally adjacent peas of eal aes wll hve the Same peak widths, and (will equal (yy. Therefore, uation 21 is reduced to aod @ a ge 3, net ae jt ching 0 = md = 10 Te Ie ve oo he tr pro om, tg ‘uno equiers ion a Sty ing enn nd 2 wld nly when th ih ot eo pnb sha Fi 37a ome et jak ep gery Se a al bse fe enpar ee eeo asic Concepts and Terms ro 4 \/ V ‘m3 Two auth pss ng heels of rena, Fo Me, Sema Cina cS “able 32 contains «summary of the most important chromatoprphic esftions and eauaton, anda complete Ist of Symbols and acronyms ined ia Append ‘THE RATE THEORY “The cast attempts to explain chromatographic band broadening were thsed on en equiv todel which eam To be known asthe Plate "Theory, While twas of son vale, 0t dal withthe nonegulis maton that ately extn the coum ad id nt addres the cases of band brosdeiag, Homever, a erative approach deserting the kinetic {acon way soon prevented it became Know asthe Rate Tory “The Orignal van Deemter Equation “The most afeetial paper using the Kinetic approach was published by sh Deemer, Klnketberg, and Zager i 1986 [3] Tented thee ‘ete that contribute to tand broadening in packed columns di So (the A-trn,Jongitodinal meleclar itfson (the Berm), and mass tanfr inthe sonar id phase (Te Cte), The broadening was xpress interme ofthe plate eight, H, as function of the average Inca ens vel, Init spe form, the van Deemer Eguaion is The Rave Theory a naman ghp newer (2) = (2) =550 ad @ Slace plate hea is inversely proportional o plate aumber small value Ingiates a narrow peak-the dstalecondidon. Ths, cat ofthe three ‘omstants. A, Band C should be minimized inorder to maximize cot ‘mn efseny. ‘The Golay Equation Since open tubular or capillary columns do not have any packing. het fate equation docs not have a9 term. Ths conclunion sam pointed oUt by Gola [i who aso propoed anew tem to deal withthe dillon ces in the gas pas of open tubular cols. His equation had 0 {Cserms.one for mass transfer in the stationary phase, Cy (sir to van| a Baie Concepts and Terms ‘Deere, and one for mas transfer in he mobile phase, Ga, The spe Gola equation is a G4 Gu) eH ‘The Beem of equation 2 acount forthe well Anown molecular dts sion The equation governing moleslareitfason i B=2De ee, where De the dition coffin forthe saute nthe cries Fire $e ilustates how s zone of molecules dfs from the repon of igh oncetetion to that of over concentration wit ine. The equation tel ‘thats salve forthe difson coefficient desirable otha diffusion | minimized, yielding a sl val for Band for 1. In general, om Atson coeticeat can be achieved by using carter uses with larger ‘molecular weights ik nope o argon nthe Golay equation equation 20), this term i vied bythe linear velocity, 0a large velit or fom ‘te wil ako minimize the contrbation ofthe Berm to the eneal pele Sroadcring, That igh velocity wl decrease the ime a sole spends ‘nthe column spd thus derese the Une avilable foe mozcla diffeion. ‘The Cems inthe Golay equation relate to mass transfer of he sole, citer inthe stonary phase ori the mobile pase deal fast ste ‘keri Chomp oepand ora Soe icy S.No ‘You Sep Recs cnneny abe Wey oe The Rate Theory s sorption and desorption will Keep the solute molecules close together and keep the tand brosdning to 2 mimi, Mass traafer i he stationary phase ean be decribed by reference 19 Fare 39. In both parts ofthe gue, the upper peak represents the ‘stebutio ofa solute inthe mobile phase andthe lower peak he dsb tion inthe stationary phase. dtbuion constant of? is wed i this txample so the lower peak has twice the afea ofthe upper ae. At equi ‘um the solute achieves lative distributions ike thos shown in part (a), but an instant ater the mobile gas moves the upper carve dowasiean sSvingise othe situation shown n(8.Te solute molecules inthe station ty pase are stationary the solute molecules in the gas phase have moved ‘shed of thes nthe stationary ise tho broadening the overall zone of ‘molec. The solute molecule which have moved aad mist now pert tin ito the saonary phase and vce vere for those that are it the ‘Ssionaty phate, a sha by the sow The fate they an make this lunsfr the es wil be the band roading "The C-term in the Goly equation i 2k CTT 25) where the average film thicknes ofthe gid stationary phase end Dp is the eifeso soeficent ofthe solute inthe sationry pha. To rnimize the coniaton ofthis term, the fm thickness boul be small 'nd the tion cetcet large. Rapid difeson through thin salons the solute molecules to stay closer together. Thin fm can be achieved by coating smal amounts of liquid om the capillary wal, but diasion efficients cannot usally be controlled except by selecting low vss ‘tionary lig, ‘Minimization of the Cerm resale when mas transfer into and ot of| the stationary guid isa fast as posible An analogy would be to coasier 4 person jampng into and out of «swimming pool ite water hallo, the proces can be done quickly: itis dep. cant ithe stationary phase a sé, modifeatios in the Cem ate neces: sary to relt i 10 the appropriate adsorption desorption Kinet Asin, {he aster the kinetics, the loser the process sto equlibum, and he es Js the hand broadening. “The ater pat ofthe Cyr the ratio hi +k Large vale of ‘est rom high sluts in the stationary phase This ratios inimized atlnge alucs of £ but very lle decease gecus beyond a kale of Stout 20 Since lg vals of tention factor esl in long analysis ties, Tite advantage fs gained by K-values lager than 20. "Mass arse inthe mobile phase can be visualized by reference to Figue 3.10 which shows the pre of s solute zone ay» consequence ofa 3 5 ; i é t i i ! i i p38 baonsing eo mat The Rate Theory 4s e310 tro mas ir nthe me ae non-ubulent Now through a tube. adequate mixing (ow Kinetic) in {he as phase can rest in band broadening becase the salute molecules inthe ecoter af the column’ move shed of thone tthe wal Sal ameter «columns minimize this broadening hecaoe the mas ante distances te felatvely sal, Golay's equation forthe Cy term i, ” where ri the radi ofthe column ‘The eative importance ofthe two C-zems nthe rate equation depends ‘primarily on the fm thickest andthe column ads: Ete [3] bs pub lished eafelations fr afew solutes on some typical 022 nm id columns. ‘A surnnay of his calculations is gven in Table 33 showing that in thin Sims (0.25 am) 85% ofthe otal Caer i attributable to mas tant in the mobile phase, (Cy) whereas fr thick Hae (50 um) its ony 3.3% Gime) pene ape6 Basle Concepts and Terms The Rate Theory ” e211 seo ey to. [An extension of hiscleltions fr other diameter eats shows that at Senaller diameters (e025 mm) the Cy term i less dominate and for lager diameters (eg 0.53 mm) its about thee Lies a large, up to sound Sb "AS poncralzation, we can conclude tat for thin lms (0.2 un) the ‘Crm is controle by mass transfer in the ebile phate: fr thick fs (2-50 po), it controle by mass raster inthe tationay phase: forthe intermediate fs (02020 pm) oth factors need wo beconsidered For the larger “wide-bore™ columns (se Chapter 6), he importance of mas transfer athe mobile phase is cosderblyeester inal, we sole thatthe C-terms are multiplied by the Heat velocity In equition 2¢, 0 they are minimized at low velocities Slow tlostie low time forthe molec to fein and out he lid pase and to fuse scrost the column in the mobile gas phase ‘Mobile Phase Mass Transfer in Packed Columns As originally proposed by van Deemer et lhe A-trm dal with ey ‘itason as shown in Figure 311 The ison paths of three moles Sr shown in the gue. All the arta the same nel potion. ba hey find dering paths tough the packed bed and arie atthe ead of the ‘column, having taeled dereatdssances Beene the fw rte fcr as is constant they ave a diferent times and ae septate rom each fer. Thus, fra age aumbee af molecules, the eddy difuson proces ‘or the multi-path elect result in band broadening a how. “The term inthe van Dest equation A= dy es) ‘where dy is the diameter ofthe partis packed ia the column and Aisa packing actor. To minimize A, small pares shouldbe wed and they Soul be tighly packed. In practi, the lower init on the patie ie Js determined bythe pressre drop actos the column andthe aby to ack uniformly very small parses. Mesh sizes around TOMID0 a co ‘mon: Small anges in sie ao promote better packing (minimal A), s0 {00/120 ise better mesh range than say S120. Since the original van Deemer equation di pot ince a Cytrm, an ‘extended van Deter equation that includes ho he Aer and the Coe ‘ermhasboenproposed{6l A simpli version ofthe eaten cauaon es avBs Skin ada a Fare De (2 where wi he obsirvton fetor for packed beds a function of the sl Seppo). This equation has found general acceptance although some thes have been proposed and are dsessed inthe next section Tt should ao be noted thatthe B-erm inthe orginal van Decmter uation iclded a tortuosity Tato, that ato accounts forte nate ‘ofthe paced bed. There no such factor in the Bterm for ope bul ‘columns of couse, Other Rate Egetions ‘Adatonal modieations to the orignal van Deemter equation hae been ‘proposed oer workers For example, one can gus that dy aiftacon {the Aterm) is part of mobile phase mass transfer (the Cartes) ot coupled wi it Giddings [7] has thorough dicated mass ianter an peters » coupled term combining eddy sition aad mass transet10 Produce a nw equation, ‘Others have define rate equations that would serve both GC and LC IM, An attesting discussion summing much ofthis wort has been published by Hawkes [9]. His summary equation i inthe some frm Gola’ hut tis ese spect. The eterences can he cnsied for mote in focmation Van Deemter Plots Wien the rate equation is plot ( v7) the so-called yan Deemer Plt takes the she of nonsymmetis hyperbola, shown in Figure 312, As ‘one would expect from an equation in wich one tetas mutpid by ‘locity while anothers vide hy there minimum in he er Sptimam velocity When provides the highs eficney and small plate ele, {Soman tie chs ema gh Pam hoagie meee a ape“8 Basic Concepts and Terms Heit at ° ©» % ein palm Deets lt Cus of Oper Tabar Clan at Cooma ‘yb Le Yong mbar oe Wy Se, Reppin Sot ey Sos Tks logical o assume that chromatography would be atid ut he (opamum) veloc represented the minimum nthe curve since yells the least peak broudeing, However, the veloc ean be increase, the analysis time wil be decreased. Consequently, chrmatopapirs hve de ‘voted their time to manipulating the van Deemer equation fo et the ‘best performance forthe shortest analy ime. By examining the tlatve ‘importance of the individual terms tothe overall equation in Figure 313, ‘one see that the upward slope as velo is ncreared comes abou com the increasing contribution of te Cte. Therefore, most tention bas been focused on minimizing them, 2 tpi hat wil be covered shortly ‘While the rate theory isa theoretical concep, taf onein practice, 's common to aban a van Deemer plo fr one's clus Im order The Rate Theory » to evaluate i andthe operating conditions. A solute is chosen and run Sothermally ata variety of Bow rates, beng sue 0 allow sien time for pressure equilibration fer each change. The pate number isevaluted fom each chromatogram using equation 1 and then ued t alate the [ate eight (equation 2). The late height aes are pote vers linet ‘locity (obtained by equation 1). The minima velocity noted swell ' the slope ofthe cuve atthe higher velocities Comparison between ‘columns wll ep ia the selection ofthe beat column, The van Deemer ‘eqaton seldom ised to calle H. ‘A Summary ofthe Rate Equations of van Deemer and Goay ‘Lets conclude this discussion by considering ony two rat equatons—one for open tubular columns and eae for packed colunas. The former represented by the Gola equation erry Sar aD, G+6ksueee ROTA DG eo ler by the extended van Deemter equation; Hand Do, AT, ot . PMT TERED, * Das ty “Ter value in improving chromatographic performance i summarized in the follwing secon Praca Impiations Returaing o our carer sugestion that chromatographer ook for ways ‘to minimize both 27 and anaes ne, let ws compare the elet of crit fason the rate equation for a capil column, One can choose to optimize "he column etitency (plate number) othe analysis tne. For» given column, a higher molecular weight gas will generate more pats since the ote diffi is minimized (Beem). Nieogen, having the higher moacular weight, shows a lower minimum H. one wishes to optimize the speed of analysis, however, itis eter 19 hoo a ighter carer gas, bike Blum, or hydrogen, Refering to Figue 5.13 one sees that iroges bas ts minimum H at linc fas veloc of 12 emisec. The minima for helium and hydrogen occur a about 20 tad 40 cases, respectively, all gases were ru at minimum H,aitropen would ‘enerate about 18% more plates, bata an sans tine 93 mes longer than hyrogen. Finaly we must examine the slope ofthe curves beyond the minimum in Figure 3.13.'We see that hyarogen, the lightest ga, atthe smallestA Redefintion of H st. ih Pe W\ 7 2 WS eee 13, ect fcr gt onan Deemer ane (025m WCU =) ‘Spies reman 2e) Heh slno Caroma, Send tion ‘evi Pca Ce" Wimp, DE Cp 8) wc Pd Cy epee ch perme Slope This means that with an increase nthe hydropen How ste, a sal toss in column efciency can be ott by a fre gan in he sped of Stagione could ehoos he column length to optimize a given separ ‘on, the lighter carter gases would provide the maximum plates pe secon fd thus the fastest analyses ines ‘As we have seen the C-terms predominate at high veloc and column ‘optimization is achioved by opimiing them. What factors eotute 10 Sh optimized Cer? Most important isthe ln hcknes which should te small: Commercial columns se avaliable wth Ss of 01 ym akhough 1035 yam fms are more common. While thin lm give high eficiencs land ae god for high-oling compounds it sould be remembered tht {hey can aosommodat only sry small sop ss ‘Sl iametrcolumnnare desirable analy, iathe Cy te). especally scouted with thn aw The smallest commercial cole are 010 fa Inside diameter (cd). Again, small sample ses ze required. Alb, we “cooing. te opin, Th te eh cr have already noted that hydrogen isthe prefered carrier gts fr a flfieent analyse; However, pei cae must be taken for safe we For packed eolsmns,thin fms, aro clos, and ela o dope carer gos ate abo deable for efcent operation. The fin thebacs ‘annot be measured easly and stan alternative, the percent by web of Tiguid phase is usally gies. The coverage Is dependent on the slate fen and density of the Sod support some equivalent loadings are sven {in Chapter (Table 53). Ift00 hte stationary hguid i aplied to the sold suppor some ot wil emain exposed and uacoeted,usaly resin Undesirable adsorption and tang, Typically colmos with an od of 18. Inch are the smallest ealsble commersnly, although socalled cto packed columns o 0.75 mum st (i6incho.d) are sve for afew phases Tn addin, the particles shold be of small uniform size ef 10-120, sest) and be phy and uniformly packed isthe column. The inet ‘ofthe solid support very important and ede in Chapter 5 ‘A REDEFINITION OF #7 Now that we have related the plate height , tothe important variables Intherate equation i might be tsful to lok it from another perspective ‘The concept of plate height originated in stilton they where colar ‘were described as continng plates o¢ “theoretical” plates, Each plate ‘ccuped a certain space (height) ia the dition column, or, there ‘were no physical plates ach egullbvium stage was considered fo be one ‘theoreti plate. Thus, the plat height was the eight of column oxepied byone plat. In chromatography we have contin the use ol thes ers. and there isa pate theory that teas the chromatographic columns ‘hough it contained theoretical pats, However, ou dacson ofthe fle theory has developed the concept of plate eight asthe extent of peak broadening fora Solute av pace though the column. Thus, «more Appropriate term might be cola deriv or rate of band broadening Toc, another definition of Hi, @ vere o isthe variance o square of the andard deviation represeat the width open, and Lele he length (or dstane) of monet oso ‘beter understanding of the meaning of ca be sane by coin ing several equations presented ear, stg hte destino 5 il to LN. Subtitting the deiatca of tequton 1) we pt2 Basie Concepts and Terms References 2 ane co) [Now we equate equations 22 and 33 e-4 on tome noe e Consequently, the meanag of for this stution (GC) i the reteation time, andthe concept of H Tor GC is est expesed aed o ee eee ova on Eqution 37 gves us definton of 1 which also provides the answer othe ‘question about the extent of peak broadening during the chromatographic roses: the peak width, expressed ino proportional othe quate rot ‘ofthe retetion time. Thus on alten column, x slate with» eeaion time tice that of another wil havea peak width 1 tines (eqte foot (012 the width ofthe other. Or, when ny one slut to compare two olumas that diet only in Teng the width of the solute peak the longer columa wil be the guare roo ofthe rato of tet lets, nes the width on the shorter column ‘THE ACHIEVEMENT OF SEPARATION Weave seen how an analyte 20 spreads or broadens sit pases hough the chromatographic column I might scem that this zone brown 4s ating counter to our intention to Separate solies and cule prevent tomography om being lective Is cumrproductive but oes not present us rom achieving separations ty chromatograph. ‘Consider the smplied equation for reslaon peste cater in this chapter ned o» Ip ALL Te Accent af Seputon ded fom dS 1 Dyas of ‘Shays are Der No Yon 8 Coy oar eth Ine om ie. M.Comangrapy Comp ant Couns Wik 8S ne, ‘wv pS Reraasoy fae Wy& Sen e ‘ile iste thatthe peak width, here represented by, increases 8 the square root af column length te Ustance between wo peak, Increase erty wth. Thue % ‘esolton i proportional tothe square oot of the cola length “Tis effect is shown graphialy in Figure 3.14 where d and my ate plotted aginst At some vale of fy indicated by the dashed lies d becomes age than wand Separation eaehieved. Our coctson i that ‘chromatography works, and Jong st two solutes have some ference in thir dstbuton constant it must be powsble to separate tem if he alan canbe made lo enough. Tati the chromatographic process lectv eventhough t produces peak broadening Tn practee, of emure, fone setdom uses increasing columt length a the only method for schie. ing separation REFERENCES 1 Ete. coma 16 47-485 98M Basie Concepts and Terms 2. ys Deemer 1, 2g 1st Kleene, A Chon Se 8.271 (84) 4 Gey ME Gat Compan BSED Dey, Boer sae ae 5 re. C, Cimaopan 1, 389-5 (0) (Som WL Aa Chm 3.0 Ot Gig 1€ Daas of Chroma, Fa bebe, New Yr. 185, 8. Koou Al Chom 20 80) 4. Stationary Phases (ofthe wo important decisions in setting up agus hromatopraphi nasi, choosing the best column (asualy the best sttonay psc) the more Important. The other, selecting the column tompertre, sles il ‘because the temperate can Be easly programmed inough 3 range of ‘ales to nd the optimum vale. (See Chapter 9) This chapter dues the tyes of sao phases thir lasication, their applications, andthe eter sed in selsting a spropente ig pate Tor given separation. With packed columns. he choice of the Stationary phise crit, bait ses ofr open ua Columns because of ther higher efcieny. Individual chapters ate devoted to each of the two column types, and tis chapter is more relevant fo picked columns (Chapter) SELECTING A COLUMN ‘This section concerns the sient has for selecting tationary phase, but fst we must admit that there are ater ways toss GC columns ‘The eas and quickest 10 ask somecne who Knows That person hy workin your laboratory or down the hal tere san experienced chrome Tographer nearby ar otrwise aeceasle to you you should not hese tok ‘There ae also chromatography supply houses wi extensive informe io, much oft already pabshed some in ther cts Tneeasagly, 55a Stationary Phases ‘spptications data ae being made avalible in computerize frm. Chron. ack fas produced a CD-ROM calles CPSCANVIEW andy Skene talled CPSCAN which contain over 129) GC and LC appa res have abo put these data on ther Web ste onthe Interact ashes Jey ‘Scetie, who has made is applications iterate aval onthe wer ‘Ask them questions gve thei pplications chemists cll Another method iso make a search ofthe Heat. GC 9 mature ‘cine: is highly probable that GC hs ready bees spied tone ‘ype of sample as tere are already over 10000 Ge publertons Weg 'eady acceso Chemeal Absractson fine, an experienced lteatoesee ‘ist should beable to come up wat suggestions fo lp sou ‘A til choice isto go othe laboratory and mate son al rns Some od columns nd typical conditions ae gested in Tale Wo ne ‘You can easly maka quik scouting run oh yout ew senile (CLASSIFICATION OF STATIONARY PHASES FOR GLC 1m Chapter it was noted thatthe stationary phase can be citer uid (a soli Liguids are more common and ge seo the mibcheoatee ‘Known as a5 iquidchromatopaphy. GLC: Solita gas sdchoomanse "raphy. OSC, wil be covered later i hs chapter, In order io se a aid asthe staiosry phan in GC, some ebeans must be found o hold he gui the column. For packed lune te ges ' coated on a solid suppor, chosen fore highsrface ates tnd crn ‘The coated supports dy packed into the eam a ah a peas For open tabula (OT) ce caplary columns the lds cea ote inside ofthe capa. To make i adhere beter. the gid phe hey ‘Extensively crossed and sometimes chemically bonded te ie feed sles sree. See Chapter 6 fo further deta. igeid Phase Requirements ‘Hundred of iquds have ben sed as sationary phases because the oly quirements rea lw vapor pressure, thermal say, and if poste TABLE Recommended Clr Sug ae 5 Teer ow ange fo San at mat) Packed si a: py ane Claseaton of Stionary Phases for GLE s SE-30 ov-210 if i Fe-At, Compson fo gd nes rm ntsc) SE poo ‘Shri VTi sgt cane dam osm be a staan tow vscsity (rast is ante. The hp umber of pos is tis made the sleon procs compictd and some chs ene ‘Send to simpy Sree il icp itsatthe fet of poy only. ‘Totwetecne wa ttonry ps hid hoch sud intro th Conpontats othe sampc tobe analed. The chemo ta “ike das ke suppests thats pol itd shold be sed sme polar analytes ada apa igi or sop ayer Fig shows ih Sepration a peste mare onto columns nope SE" tnd amore polar OV-210. Gary. he een of th proper soon igi vee npotan in the ca por clu woe wel for penises "The nonpolar E20 a god ctu Chih elfen) Cri Dea chee rma, oa koe hmm (aah ese) fn comprio owe datoay plac whih hai exten dierece ‘npolniy. the ode afeaioncan etna rene For cample Figure {Zahn te separation our compounds tha ave sia ling ps a beth a pour coum, Carbowar SM ands nonpolar noms S30 {i} The cton oer evescd. The eta chanboy atoms phase ov dig aera pans fw OA aly cy Cem “Cason tbe ade tame Un as Coy, et pee38 Stationary Phases by Wy ‘ Mi 42 of atone pty 0 amps prion) Crone 8 pt) (C20 ropa Sap tng ps epee {Bren 0) Som hae Reged ont Play not always so dramatic, but investigators shouldbe aware that ‘large change in column polarity may esl tna change in eation eee. Fale to contr the indviual retention tines a series of sluts on ‘column of diferent polarity could result in misidentitions and serious ‘rvors in ana, Clasifeion of utonary Phases for GLE 9 The chemi’s problem isto pedi retention behavior for slates wile Jacking # good system for apecivingpolaiy. We saw in Chapter 3 that the adjusted reenton volume i diely proportional othe distebation foesant Ke, so it could serve as a measure Of polarity, but dstrbaton| ustants ae not generally Known The best we cat do within the conten tis brit text eto aco some of he bei principles of polity based ‘on ou knowledge of intermoecalar forces. Polarity and Intermolecular Forces ening the polarity ofa stationary phase is complicated and aot easy ‘quantied Polarity i dstemined by intermolecular forces which are con ‘ex and dif to predict in chromatographic stems. The polarity of a ote igudcan be spectedby is dipole moment. Other pha properties, ‘chasollig pont and vporpresiure, rele the extent of ilermoleal forces. lage dipole momeat anda high Bolg pint would vetect high polrtyand song ntermolecslar forces, However, hve parameters late fo pure ligand in GLC, we are interested in intermolecular forces betwoun two diferent moleeles—a sot in the vapor tate and gid sainary phase Such stem i complicsed and ti impose at thi time to produce a single aumeria cle that canbe used to epson ll, posible interactions. ‘ssl, intermolecular fores have been casi a an der Waals {forces listed in Table 42) and hydrogen bonds Of the van det Wane forces. depersonspreseat between ll organic compounds even nonpolar ‘ones Consequently. dispersion spot of muh interest exces when nop lar hydrocarbons are the solutes. Induton and oicataion forces Bre selectivity to chromatographic stems and they eause the polarity we have been dsasing, However, attempts by chromatograpers to refine these ‘everalations of play oto wore tatu parameters have wot bora of ‘much paca value "Hydrogen bonding is better understood andi evidenced only if on of the molecules has a yrogen atom bonded to an electronegative atom ike titogen oF oxygen. Examples are alcohol and amines hich can both donate and receive a hydrogen tom to form & hydrogen bond. Other Imolecules such at ether, aldehyde, ketones, and enters ean only accept protoas—ihey have aoe to donate. Hence they ca frm hydrogen bonds nly with donors sich as slohos aad amines Hydrogen bond are rela Td inte amines ie. ian Bittefa Stationary Phases ‘ively strong forces and the te very niportant in chromatography: prt ating molecoles are usualy cased as hyarogen bom eens nee Fyropen bond acceptors ‘The strength of hydrogen bonding ca abo cate unwanted interactions Sots capable of hydrogen honing can become attached to the was of injection ports slid support and column tubing. Often thee senorton, ‘est insow desorption pings to nonyametnieal peaks calle ing ‘peaks This undesirable esymmety in peak cape can ion he eenoned by deriva the surtacehyaronlgroupron tic walls andonaldeeenee surfaces Slantzation of sd suport dcused in Chaper 3. ‘The combined effet of alinermeclr forces cannot be teed theo- ‘etal to producea “plait” value for aiven oleae Rather compar cal measurements and ints clelted ftom empincal mencucernts have Been devised to represent molecular play Separation Factor ‘The separation factor, a: i a parameter measuring relative distbition ere Vth speci retain volune (he et retention vue at TC pot pram of stationary pas). When benzene and loherane are ‘Sromatopaphed on dinnyphhaate at °K. thir cy cots Seton tobe OS? aod epetvely [4 aad = 1.8 = O8DUSE SBeazene retained mre tan eobexae byte pla nonstate ‘cnr ofits large intrasseusrinteracions Whie say Sens [Meno commoaly determised or hs arp, csr ht eye 4 ‘ald mean for exreinginermalecuar ater LIQUID STATIONARY PHASES (GLC) Sualane has already een discussed asthe ligud phase considered to have the Teast polarity, Isa saturated hydrocarbon withthe formula Cy Has, ite sructare i shown in Figure 45 ts upper temperature lint i oly 12°C soa larger partin, Apolane 8, withthe formula CH has often been sed asasubstiuteveathoughitsslahty more polasee Table 4, CH Ch Hy om 1 1 i HOCH, JH {CH, J CHOM, CHM CHO CH oH CS ch Ch AS, Saunt ety ced Conoe Stationary Phases Liquid Suatiomary Phases (GLC) o Silcone Polymers Stone polymers have god temperature stability and modied scone olymers now domiat the commonly wed lid pts A range of Polaris canbe proved by changing the prentge of pla grap ar ‘tapi pee and eysnopropsl rope Pe last pour dines one whose struct town in Figure 65s ot under the nase ‘0V-1 and OVC 101 by Oto Vay Specty Chemical, he former be 2 gum andthe ater «bg Both are ince Ins complet sting of Sion phases in Appendix VI eee rom te MeReyols constants (Table 4), it ca be cen hat OV nd OV-01 have ential the sme platy an ae sgh polar than Apolne #7. AS the met group re roplced ty the more plat they and eyanoprpyl groups, he plates incase idence y the Increasing McReynos constants Table 4 ss some alerted tons se for these popmers by other maneacres ‘Other Common Phases "en this fon listing of scone polymers does not meet the needs of all chromatographers who seek gus with higher polarity andor higher ‘operating temperatures seis of poetlenglyol poner hs et some ofthe need for higher polit see these mater cn byrogen bond. Te stature of these poets piven in Figure 43. The aprox. rate molecular weight js given as « numeri value inthe nate. For ‘example Carbowax 20M hat a moleslr weight of 20000 she highent [FABLES ga Soe Pe gl Phe ovemn0 seam pew ge Oncor) 2. Setar of Coan #20 «pres petal i an aenge Mictir mig 3090. molecular weight commercially avaliable and canbe used upto 25°C in packed colons and 280° in some Bonded capliry colts. “A series of exrborane scone polymer (se Fig. 48) has ben designed spec for hightemperature work. Thy were fit synthesized in 1964 ty Olin Chemical and ae so under the tradename Detsi.® Det 300° ithe leas polar, having all methy groups in the ein It canbe sed vp ware Recommended Stationary Phases For practical reasons it is desirable to have ony the minimum sumer of cts that will sve one's most fequeat separation problems. Open {bul columas are so elie! that ewer of them ae wally needed bt {tiscommoa io have two to fou fren phases and sever diferent im thikpesses and lengli. More spec information piven in each 0 he ‘especie chapters on packed and capllary clans (Choosing Stationary Liquid Phase 11s clear from the foregoing discussion that no convenient ste has been found fr guiding the sleton proses. Certain one cannot rel | “MeReynolds constants alone. The simple maxim commonty ied isthe ob with which we began this secon "ike dave ike” Which 9 ‘ne chooses nonpolar column fore poapoar mixture and polar esl foe polar mito "Aneception this generalization ours when one attempsto separate similar solutes suchas ores. For example, the rlene omer reall, ‘more of les nonpolar and have ssa boing points A nonpolar sation FAS en 8» Stationary Phases ‘ry phase will not he saisaciory for thee searation beens they do not ‘ary much i either boiling point or in poly. To aceatuate the small, ferences in polarity reguies apolar stationary phase like DB-WAX® Figure 49 shows «good separation of thi challenging separation, Following the general polarity rue and wing packed columns gly loaded (5), fou-o01,2 mm ia, OV-101 column would be a god choice {or a nonpolar sample and a similar Carbowax 20M column for polar sample, Between these vo exttemes, one a the slicone polymer of ater ‘mediate polarity (suchas OV17) could be ied. Other spec packings ae diseased in Chapter 5 or open tubular columns th choice of stationary phase is ch less stat thin fn (025 ym) 1Ssmeter methyl icone column (OV-101) DaMAK30 mete x 0250 10. Pe 49, Spat of ne tnt pr lan, DB-WaL® Coe of W Sei Ie un er 1M cmap Comp ut Cnr a Wy Som Rew Yr Wp. Remo cry of ke Wil)’ Sos Sod Saionary Phases (GSC) n foes Ctopa Cag Cech Grphpet fe ce Gon Cray Ses vould be good for general screening sir, but more polar scone Polymer (fr example the cjano-detivaives, OV 225 or OV275) would Bebetier or more pola samples. The capil coluns eqialen of Cabo- wax 20M (such 3 DB- WAX) is logal choice aso, ven thouph these ‘columns are easy oxidized and have relatively short elites ‘final consideration in choasing gid phate is emperatre lini tions. At the upper end temperatures eached where the vapor peste ‘ofthe liquids too high andi bleeds off the column pvngabightockpround| ‘eter signal sich high temperatres the column heme shot and ‘he chromatography is poor dust the Bleed, The tales of common lid [Phases his chapter have icluded these upper temperature nie for he ‘hates when used in packed columns. The limi for open ube eure esimilar~sualy bit higher bonded the column, The lower temper ‘ture imi is usually the fering pont or gas transtion temperature of the polymer. The asic example ix Carbowax 20M whichis sold at ‘oom temperature apd melts around FC i lower temperatre limi SOLID STATIONARY PHASES (GSC) Sols wed in GSC are wadonally run in packed columns, the subject of the nest chapter. However, st of commen sii given in Tate 46 [As wit the sll suppor used in GLC, thee solide should ave sal pate sizes and be uriform—forexaspe VID mesh range. ‘Some af these vol have beta coated onthe ine walls of capillary G* can only hold small amounts of quid (ply 35%), Low loadings are Better for high efficiency and high oiling compounds, andthe high loadings are ete for large samples or waa slates ene {ocexample A soltion ofthe stationary pase made a vlae when, mined withthe sod support and evaporated to remove the slvet. The nal material even those with 258 gud stationary phase (on Chronos 'P®) will appear ary and wl pack cay into the olen, LIQUID STATIONARY PHASES Virwally every nonvolatile liga found in a common chemi Iaboratory hasbeen tested es posible stationary phase As consequence, there ar oo many quid phases sted in commercial suppliers catalogs (peal ‘shout 210 of them) The problem i forest the long lt of pases To {ew which wil solve mow analytical problems To this ed soveal workers have published thei Isso peterced phases. A few of thexe coe ae Tinted in Table 54. Tn genera, they include» nonpolar column ike the Yan Xa te aoe SRT Nt WALA Pry a sen Ce Soy,6 Packed Columns and Inlet Sold Stationary Phases (GSC) 7 methyl scones, several of intermediate polity,» highly por scone like OV.273, and a polyyeltke Carbowax ‘A secondary consideration the amount of stationary phase needed to atthe solid support Table 51 listed the oper limits or sane sports. ‘The lover lnit usualy the miasmum amount tht il give complete coverage of the suppor sacs, an amount thts dependent onthe wee tea (aio listed in Tele 1). However, uiform coating are diel fo iain especially for polar hquis, aad the minimum percentage usually Aetermined by ial and error. ‘tire consideration i column length, but itis ot rita ifthe instru iment i capable of programmed temperature (sce Chapter 9). Column lengns are wualy shor (to 8 meters) for conveniences both packing ted handling, ‘Choosiogthe best column iid phate) fora given sample was dcised in Chapter 4 but a usefl etorene providing neatly 20) examples fatal Separations on packed colums bas been made valiabereceniyby Supa {2} Other saplers aso provide sppbcason information SOLID STATIONARY PHASES (GSC) ‘Common adsorbent soi ike sca gel and alumina are uke in GSC, bat ‘most the sallds wed stationary phases hive been desloned fr spect {ppliations in GSC. Table 6 Ise some of thom and ths chapter wi irate some of thee common wes, ‘A typieal separation of xed gases on sis gel is shown in Figute 51 Although he pen shapes and ple number ar athe goodin this example, many ofthe ois used in GSC produce poo shapes (eal tiling) tad ail characteristics nde properly coupled tothe ctomalopaph Some ‘afte are refereed to as mass selective detector (MSD) heh nde at they ate considered Ge det mas spectomtr with 8 as ‘shromatograph i avery powerful ase, and popular combintens ad itis treated more detail in Chaper 10, Fs more information on these ad oer decors in Table 7.1, consult ‘he references piven nthe table References [REFERENCES pre ete eee ee eee eee eeerenieratnen eee pee ee eg ie 3 ie At Ch 4 819-29). pesca nate erence eae ie She eiasares are 1m Tada Ot Choy 07 (0. oncorsoncmanes pope dee crtenereateaes Sones cm eee 124, Grima FP, Daca for Cap CArmtopraphy HLH il nd DM, Roe anna een cee Giese8. Qualitative and Quantitative Analysi IS x: como can ead for bth eine ad untae Aa: Beas more wl for quae nas Se ls ‘Sipe devoted th top: homee it Bop wi et oak a ‘qualitative analysis, oa = ‘QUALITATIVE ANALYSIS, ‘Th chromatographic parameter wed or usatv ana ithe ten ton volume or some cl Tele’ parte: However nee enon Parameters cannot confirm peak deny it common to coupe s man Soecrmter (MS) tthe OC (GC-MS) for quale aay GC MS ‘Foo wide wed tat itis dicted in eta Caper ‘Table 6 Tas the most common ethos ined for quliative ass in GC. Reerence 1 god sua ofthese and oer metbose Retention Parameters The retention tine for agen sole canbe wed for idea {allowing oun ari re ep constant eh sary ss tiki sig) temps ere arp a 126 Tie MSD faa eke tc) 2 spear otra 1, ‘Mar ei wor deny ne) Hue sate). As an example, consider that an unkown sample produced the ‘hromatogram showa ia Figure &la If one wished 10 know which ofthe ‘omponents were nalcohol, «sees of maleoba stndueds could be run Under ential condtions producing chromatogram ike Figure &1b. As ‘Show in the gore thoe peaks whose fetetion ines exactly match hose ‘tine standards canbe ented athe mlcohot Tas process wl oly ‘work ithe components ofthe unknown are alcohols. "This procedure wl not be effective ithe numberof possible compounds Islarge retention volumes arent that characters. Snce here are over 5.00 organi compounds in common ts, the gus ebomaogzaph by Suet annot be wot enya singe compound rom among thar group. Retention times are characters of OC system, but hey ae not uigue, S0 GC retenion tines cannot be use for qualitative confirmation. owsver, relative retention volsmes are more reproduble tha individ sal tenon volmes so guntative data should be reported on a relative [ss The retention index stibuted to Kevat (ce Chapter) has become {elabe method for porting retention data. Ia retention parameters {orbe used for usllative eatifeaton or dasa, the Koval eer- to index s+ good method To choose Selestive Detectors and Dual Detectors Sette GC dees a otis be wed lp ety ac empound to wich they show Bgh senity. The It feels i ‘Ghaper Team becomes fr farther intomaon and vtresssQualiiacive and Quantiative Analysis se ae 1 aa i i i | i pA, eniaion at woo by tn ine gan Fm Mle, hme Compa Conan Wy se Seat 6 ero ory ata Wy Sn More interesting is the use of two diferent detectors in parallel at the ext of» GC conse eal sal channel detection, The detecton ‘hosen should ave major ferences in sensi fo ferent atscs ot ‘compounds. Both signals are recorded simutancousty producing parse ‘hrometograms ike those shown in Figure 82 Tdeaueatins canbe made Qualitative Analysis Be 482. _Dan dune penton of Cans of pin sample ute DCI8 ‘Show chun ocho up Tom ii CSomamraphy Co nd arn a Wey & Sos, Rew a, 9 Reprod oy a oe ‘Si Smt ‘by inspection of the chromatograms (Fig. 83) or from the ratios ofthe lem Ms hr. BM Ae nha, Vl 3 wn, 1 Sa 1F, Dein) an Sor, 1.67 Crag 8 243908. 1 Hu. Ooomanpapy, Vo 23.1. ing Ea, Deer, 8 18, ‘Gye pp a 12 Bene, Din Je, MPa Dh, Wee JA Lav J Am ESS evan 1, Nato Cron, DR, Hi eM, Anal ha 8,20 98), 1 iin CL Seer 2 3990) 1 dog aA Chm 28-20 (8, 1, ar BJ Chom Ba 5,29 (580 1 Mao E, Ke Cal Pan Ca 1 Choy, 15 (989. Programmed Temperature Programmed temperature ga ehvomstography (PTC) isthe proces of increasing the column temperature during 8 OC ron Tis very elective method for optimizing sn analysand i ofen sed for secening ne ‘simples. Before desing it in deta et us consider the pera elec ‘of temperature on gas chromaograpic ress ‘TEMPERATURE EFFECTS ‘being the nature of the stationary phase. Reteation times and retention factors decreases temperture crete besase the dtbution constants ae temperatue-dependent in accordance with the Clauss Cpeyron auation, tog =~ 8 + cont o where 42s the entsipy of vaporization at abolte temperate ; is the ps constant and he compound's vapor presure at this tempera ‘tre, The equation indicates thes to the {abentuts) temperatare decreases, eee eee — the vapor pressure of the alate dees lori A decrease ia ‘Spor restr eas nets ie reauve amount lt the Trobe pean nese ihe eteaon factor, a a ese itreteaton tine Fre 1 plot of the log oft retention oe esi To afew cota sls Hate ti eatonsip. Sag ns re ote over ‘Tne temperature range nasorane ni our prediction bse on aston the dpe ofeach ine propetna to tat se ently ‘eporatin an ean be asmed be sonstaat over the femperatre fo a fst approximation, the Hine in Figure 9. are paral, nde that the eainpc of vores or tes compounds rene the fine A ser specton eve ha any pio ics vere spy eS. Tenure deed fn ame Reprint peman tom. ‘Spy 500 Rope for Ll om er M2 Chomp oer a cen wy Sn Sov arp. 1. Regd an an4 Programmed Temperature tow temperatures. From this obrervation we an draw the wl genera ization that GC separations are sual Beer a lover temperatures BL too at the two solutes, wacane and m-Aeorotleee: thc nes cos bout 40°C. At that temperature, 1°, they cannot be seared tt lower temperature the toluene eles frst Duta higher emer he revere tre. Whi mot common for elton orders to eves, ‘oppen resuing in miseataton of peaks See fr crample te work ‘of His with chlorinated pesticides (f Below ist ofthe consequences of ieeasing the tecpeature for OC ana Effect of Temperature Increase + Retention ime and retention volume decrease 1 Retention factor deceases + Selestiity (a changes (wsalydereases) + ieieny ()ierease sihly “The eet of temperature on elieny i ite complex [2] a docs nat lwaysincrease Usually isa minor eect and less portant than he elle fon cluma thermodynamics (selecuvity). Oval, however, temperature fffets are very signifiant and PTC i ery powerfl ADVANTAGES AND DISADVANTAGES OF PTGC ‘a sample being analyzed by GC contains components whore vapoe pres sures (bling pins) exend over a wide range, ite oten impose to Select one temperature which wll be suitable fr an bathe fun. AS a ‘xample, consider the separation of a wide range of homologs like the Kerosene sample shown in igure 92a, An sothermal run at 180° prevents the lighter components (| DETECTOR vacuum [_wass srecTau OAT cL [_ | me FAM, Sch of ma estan GC-MS img ncnee ene| >| Ey |] ner jmocorero| | seonyoret toe ng Comping of G10 cnr wins ae ma oo a meaty renege cee ni charges mee ts soa menba en soredtnaeaea moa Sci ae arate pee seit fc anced meen ee emt ad cinen es naive cue St Ses wheter anand intdarnceretag spamenomte eater as Beyer gerne Pee meses om eatin da pa ne cts a tm To ‘A) Capillary Direct interface is sous Soom |_Heste Taneie 1B) Jet Separator (Packed Columns) Yeon ——p =) FL cot goo oem Fe MS GC-MS etnfeed Special Topics ‘Oder GC-MS stem aed packed clus, wsully 2 mm id, with hw ates of abot 30 Lin These packed clan ot eed an Interface ite thet sparatr shows Figure 1035. sopare so Sst of las tubes aligned with sal tance (~ inn) betncen thc ‘Mos ofthe cai (sly Ho entering rom be GCcol's pmped sy by ssa rst em The nts olan thee moment and pas preferential the second capillary eit ‘he MS sure, Sample earcneat xr nd the nial stro pre sca rat ed slowing Santander fate Both temperate an sac aiy othe Base Separtr mst ‘cart oto tho maxi tame ans an to preore Spl ite Tontzation Sources ‘Analte molecules mst stb onze in nde toe traced (r replied) bythe proper magnetic or electrical felde Tete are aumerovsfonzation ‘techniques, but electron input (Ei the oldest, most common and most ‘smpe. The ionization soure i beated and unt acu somos senples ‘ate easily vaporized and then ionized, Ionization i wally accompined ‘by impact ofa highly energetic (70 ev) elecuon beam. ‘Apical sure & shown schematically in Figure 104, Etunt fom {te GC column passes into a heated sniation source at low vacvum, letras are drawn out from a tungsten lament by oletor valuge of ‘Mev. The voliage applied othe lament defines the energy ofthe elec ons ‘These high energy electrons strike the nesta analyte molecules cag cesing Elects || = “To in Tl y — 6¢.cotrm (ove Sams cael) vet's) Ccoector (oe) Fe 9A. Becton ing (1 sue GC-MS 1s7 ‘onion (ususly lo ofan eestron) and fragmentation. This oization {echnique produces almost exclusively posive fos Mb eM 26 o [Aternate means of achieving jonization include chemical ionization (CD. negative chemical nization (NCI), and fast atom bombardment (FAB). In Cl reagent gas like methane i adtted tothe ion camber there i onined producing ao that undergoes furter reactions 0 proce secondary fons. For example: Che CH He e CH + CHa CHE + CH, ° “The secondary ion (CH in this example serves as a reopen onze the farple sent, Usa ths proces est nese frepmentation abd more Kimpe mss spectra. The sajor MS peaks that normaly rest are (M De). (Me and (M29), where M6 the mass ofthe analyte being tied, "To perform chemical ionization the lon volume ofthe spectrometers sual eiferent from fhe one used for El the operaing prose higher (Garialy doe to the addinal reagent ga), and the temperatures lower. {Cenain pes of molecules also yeld pod negate tn specta by NCL providing anther option for anaes. However, mst bench-top GC-MS [suments are not capable of Cl operation. "A comparison of CI and El spect s shown in Figure 105 fr ora. 2 ‘ariturate witha molecule weight of 240, Te base peak inthe Cl spec trum is 241 the expected (8 1) peak. Thee ate some other, sal ‘peaks ut tht spectrum shows the value ofthe CI method in providing fr acignment ofthe molecular weight. The EI spectrum, on the other In shows avery smal parent ion with major pea at 1a and 136. These fagment fons can be sed to aid inthe assignment of the strctere— Information not pended by Cl Analyzers and Detectors “Aferioniatonte charged parle are repelled and atracteby charged Tenses nt the as analyzer, ere the sai pels ae separated by it Inns tonchage rai (mi) by eter magnet or electric il. Typical thas analzers for GC-MS are quadrupoles orton aps. Other analyzes re singe focsing magnetic sector, double focsing apneic scar (high ‘esolton, more expensive), and tne of igh. "The quadrpote as analyacr consis of four hypebolic ros tight anges to eachother (eee Fig 108), A DC voltage applied to al rds {Gdhacent rds ave oppose sia) aed the sins he voltage ae rapidly‘Special Topics P33 CL. a i: a i: = a a ab we mm LI: “lm Eo piu. 2 oe mw mae e105. Conprton eft an laps ol OW = 4). foyer Arai tequecy is ako applied othe four ro. Depentin {nthe onto athe ai ect and thet crete ‘ons of only one mas-o-charge ra wll ps trou ne eso ey {he detector. lon wth ether mi ration wil strike the ros and ae i srke the rods and be ann _y aa Quadrupole Fe 184 Ouse mas mje (GC-MS 159 “The quadrupole analyzer has the advantages of simply, smal sie, moderate cost snd ead snnning Which make ideal for GC-MS stom [Hisrertedo sbovt 200 Dalton and haslow resolution when compared to double foesing mass spectometr Figure 10.7 shows schematcaly anion trap mass analyzer which was developed special for GC-MS. Iisa simpler version ofthe quadrapole {which the ring electrode, baving ony a raio frequency sped to ‘eres eset as monopole to define astablerezion for chaged species inside the cour electrode space. Tere are wo end cape onthe op and ‘bottom the crulr ring electrode. Etfuent rom the GC enters the top {nd cap, some analytes are onized and then trapped in table wales imide the rng letrode The radio requeney can Dealteredo jet seques- "aly fos with selected i ats om the on ap and pass them through the end cap tothe detector on trap ae ako spe in design, modes in cos, sd capable of rpid setnaingfor GC-MS sppations. The spectra generated often die rom ‘asl jondroplespetra, and some ions may undergo dsocaion ot fonfmoleule colisias inside the fon trap. "Ate separation of he ons produced detector, usually continuous <éynde version of electron multiple, i sed to count the fons and enerte a mas spectrum, Such a detector i shown schematically Figure {us tons tom the mase analyzer strike the semconductve surface and {eles actseade of electrons. These are acserted bya potential ile ‘nce to mother poron ofthe semconductive surface where a arper ‘hoade of elects tess. This proces i repeated several mes unt “plicaion of the orignal wesk pst # mapniied about L-milion ‘ote ta the entre MS ytens der high acoum This ra een requirement to avid the lus ofthe charged spores by calision wih other fons, mo ccules, or surfaces. "Te mas spectra is simply 2 pot of the fo abundance asa function cof mi. Under controlled condions, the rates of fon abundance and the ENOCAP LAMENT ELECTRON ‘CURRENT FING ELECTRODE | 2 sons execrnon “SON, REE oooe 1g 02 loneap maser161 Special Topics GC-MS Conductive Sursce Capabilities of GC-MS, gle fon cone {GC-MS combines the advantages of both ecnigues: the high esol¥ing power an the speed of snl of GC retained, whe the MS provides > otk patie eaticatn and quantitative analyses dowa tothe piv e Mass ranges of 10 t9 609 allo ae common for ow cost systems, nd a ve Stoo Upto 100) Dalions for more expensive systems. Coss range fom 40K eter Fe t8h sec utr (omit ec specie mz species presen ate unique foreach compound. They canbe ‘sed to establish the molecular nepht snd the cemal sauce of ‘ach compound Hinton, Alter .1-Thompson wea mas spectrometer to sear ti in 913, MS was slowly developed snd snproved aan analy oo roved vo be power for senting unknown Compounds s wll ‘hciding scares ofboth inorganic and organi Smpocnds. It mas ‘ly used forth caracterznion of pelea prods and prot Tous ve own nn mor aay HCC nt en noes [MS waist couplet GCin 959 by Got [6 The eat nstrments were expensive cumbersome and comple, ly eqns cosderate pers and mastenance to kee them runing, Magne sector ae tents were the most popule typeof sna, howener ey oul nt Provide the rpidseanaing (ew scons) secs) to encate 2 ma Secu of «peak citing fom a GC: Far soning nays woud ereaured By telat 196 was obvious hat GC mas use ana market and growing api, bt no GC detector roiled es th iomaton ‘es avalabl om MS. Thos was bors he deieated “GC MS" stems ‘heres new MS was designed a detest forthe Ge By speciing iat the mame int would few GG. the MS egiremens could be simplified. The mae range cou be ined to abou 600 Datos tow resolution was satstatry hese the GC psssued high elton ‘abies so the eluting peas would in monte be “pure The tough fat was the development of rp setoning Seite (heal 40-40 Baton several times pe second) sd more spl, ragged istramcns ‘hich culd be tne in oui aaa! laboratories The quaapaa, Sod ater, he fo rap esgn ttha ed. tp to SISK for simple beachop stems iitton of GC-MS Systems {GC-MSintruments re a capital expense item: they are more complicated {fo operat than a GC. ad there it lack of sled GC-MS operators Fow cobeyes tun student on GC-MS ystems due to both lack of pes foreach purpocs and the lick of expertise of many cole professors. Data Anais ‘A ypicalapillary chromatogra of hydrocarbon sample run on GC-MS fas the same appearances woul with an FID (se Fig. 109). Note the ‘arrow peak wis ypealyeround I second rls a half height THs Icens thatthe MS stom mst sean the GC. peak about 10 times pet ‘Sond onde o eta good mss specu. gute 1010 shows x proposed mechanism forthe fragmentation of hexane (peak 4in Fig. 109) i the fon sours ofan GC-MS sytem. An Clectfon sks the parent molecule, ejecting one elton and generating the molar fon (vz = 86). Ths species a0 stble, however, ad Fapidly decomposes to more stable fragments inthis case ml of 7,57, ‘Sand 20 Dalton: That frogment wi the highest abundance, lz = 57, TIME (riny ——> * 100, To in camara 11) of yucion is162 onereccnenery me = 8 fa eee ne: tosonamy sora 4 ca — 4 Prenat CouenenoHfan* cHcH.cHoHs ONO Ghent setae p10 wopmeren) 7 M884, Fragman oes na wa ene scaled the base pea, andthe data sytem plots as 100% ofthe spectrum Seale. Other peas ae ploted relative to the bse peak, andthe res typical mas spectrum of mhexane (se Fig 10.1) ‘Daa can be plotted in two way ether a5 total can (TIC—Toal fon CChromsiogram) ora «smal number of indvidoa ons (SIM— Selected Ton Monitorig) characteristic of parclar compound (ee Fig 1012). ‘A total ion chomatoprany i wed to idly taknown compounds, A ‘pected mass range fe sanned—for example 40a 400 Daltns All peaks ‘re vepored so the mas spectra can be rtived from the ome and co MOL We 1 a7 we Fe ML Man pcram Rete ep iin Coparvon ttn amaogm (TC ad eke oa sna, te wed 1 identi ech peak The comps data se rapt comptes ch unknown mas specu with oer 15D referee spect 6 iar lea Matching peta rogues ote seconds wi the ate Anta opstns thong th Gerd guia nals The data eqison fate neceaay to san alos in he sted rage owe senate) Tinta an! usualy quantinton ct optimal of data poi) inslced fon ontring however, one small amber fos (9p cally os mun ar6) re monitored There 2 ter data acgiston Fle fete of te GC penk (=I scond, so quantitate da eter anny arent mprone SIM cannot sed for quaiaine alec (oral mses ae acaned), but the Pest mode Yor ace rata of urgeted compound fen doa othe pb ve “Rowe yr ote on esp allows oe operated in soca way inate onal agent frm thlonzton procs ate gun exposed 0 freee ar molealescauing secondary fragmentation frm the ‘Si infused dssosation (CID) Theres snort hat etained {hy dal stage ms spectomete sual ttered to MSM) and provides ren higher eect Tis mode operation Scaled sled Feacon onitoring (SRM) aad ir walale fo some newer ion Wap GENS facut CHIRAL ANALYSIS BY GC Chiral separation cither GCor HPLC san essential tpi the yates, Shuracenation and the wtztion of cial compounds (rs, ese. favor phoremones ee). Asa understanding ofthe sinc of chia. iy on boat aetiy increase, legislation repuating chia compounds becomes mre widesprad and stringent an the need fr high resolution ‘Separationtecinigs increases, Chiral separation by eaplry GC provides164 Special Topics high tency, sey. nd speed of aly bt nied ty the ed fo vty. Combiang ra aes nto peoranes hs sed a Increased temperate sy “GC separation of ean: cn be peromed citer dct (of a ciel aoa pe, CSP) or de (ann tomveian ‘strcomer deratnes an separain by nn hal anny pases) ‘The dew method» precede fing tila miming sce on venta Be Peon ol am oe ee “iL oom one eh — Boho aye oats St a mors) San =o = = ma one — "i = enc omee se Secs G ve ' | ie aa, iting hou af eda Ti dein _ say acer Napoca ty tt peat caet F {Sng scsamonree repens ns Sean COUNT etn ts __ meceernin oan, a ee sation ae din he Rees (2-2 Pee em tune aun ps tt apes opts : Lattevid “The reactions to produce volatile derivatives can be classified as silylation, ‘fompnooce acylation, aikylation, and coordination complexation, Examples of the frst c eee . as ala yogis ey se _ — oa eal ace tacos crag any ipackpeinacnacte raisehiocet han toasts oven Cae ter nareenisteue aes > oF not. A review of this subject has appeared recently [29]. Toc Teicuarecne ooaeatnomne Ped vth atl, andtpeurgennne phaoesrphscntd enna yucnen eae sy Ber aioe tioned ses ee eet Dore oF -,rLlrtrtt Y- vate ote ec do we et © Introduce ‘he trimethyl group nt the anaite to make i ole. A