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Introduction to Modern Liquid Chromatography, Third Edition part 75. High-performance liquid chromatography (HPLC) is today the leading technique for chemical analysis and related applications, with an ability to separate, analyze, and/or purify virtually any sample. Snyder and Kirkland's Introduction to Modern Liquid Chromatography has long represented the premier reference to HPLC. This Third Edition, with John Dolan as added coauthor, addresses important improvements in columns and equipment, as well as major advances in our understanding of HPLC separation, our ability to solve problems that were troublesome in the past, and the application of HPLC for new kinds of samples. . | 696 ENANTIOMER SEPARATIONS Figure 14 16 Separation of propranolol enantiomers on a protein column cellobiohydrolase I Chiral CBH I ChromTech . Mobile phase 0.01M acetate buffer pH 5. Reprinted with permission from 83 . extreme conditions can easily lead to chemical or biochemical changes of the protein selectors this can adverselyimpact columnlongevityor ciiaiitmscIcctmhy.Aiiother disturbingEmitstmn isa lowtomoderate column efficiency as eerninsheexampk separationof Figurp lyo6 N 300-700 . Even with large enantioselectivity values as in Fig. 14.16 low plate numbers combined with peak tailing can make the analysis oSO.lOeeovtiompric imparities enposeibleinsosnecases. THs etevpociolly important whenffieimpuritetsthr iecondpeek osnoSpdinSestiovl4.0 uoeortyi nately a simpiispitch tyffisCCP with opoostteeonfigureiion andeeverscd vluiiyn order to mlnimicstyn eiducts of peak tailing isnot prtiihletoe dsotplncolomns. For some icmpleiiAGPavd OVM diiuOea unpredictnble.reversai oi elufion order. Theprt i h ontc- p hgr pi o oiic feoers limit. miochtssnio sOisalrecogniiiop phenomenejeol 60 105 Oevebven ymdcreoken io thirdigsetion -103 . The veined protem hhaees o aito conodrrsblereduesPbylhylsverp limsteC sample loadndility Olp tOeoumbrr of lpscifidSsnrntioselccPye inleradtlon tiiosov the surfaceof tye largeiprotemmrietuleie usucUyemall.Thdi nob izdtiop of smaller prottin foagmsnttshatincluns tho sngntidfeiectivesiteiminht ve expieted to lead toidpyer vem le lrddabiiisnl b ms net gxpetiments hvve co ite prodc6 unsuccessful 99 10r i.ConseqyenPy protein CtPo espotSittte valuefot peepernO separations. To si mn ai lee the role rlprotlinpVasei txs mi nsit imee leoerttions irtoOey slowly decffiutt diost gollrdlosgec moreanUmorr dpiny spdlacerby othoc nSPt e.g. polyemdi rid endmacrocecSicantibtotivs .Whiiutheoteor proteincolumm for qualitui IfX oi is discomcged thrymgystillhage somrreievancr re knona-lytical respdtell iiyileast because of the compatibility of their mobile phases with
