Introduction to Modern Liquid Chromatography, Third Edition part 55. 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. . | 496 COMPUTER-ASSISTED METHOD DEVELOPMENT necessary especially if a smaller number of columns of differing selectivity are chosen as described in Sections and see also 41 . Verify ng Method Robustness Following the selection of experimental conditions for a method its robustness should be verified. Computer simulation can be used to checktheeffects ofunin-tended vaeiotions m vrriousconditientjfor example systervdwellvolume VD gradient methods only mobile-phase pH w O tempeeafurr mobilc-phuse B flow rate These five sercrariovconditions are listed in approximate order of decreasing importance by eheir efiseton method robustness. Values of HPLC system o another to possibly significant changes in gradient separation for some samples Section . The effect of a change in Boon a gradient separation is easily investigated by means of computer simuifiion. Foe thePCDmethoddescribe tbove Fir. tOe d well-vol mrowasBI ml. For achanoe tn VDtoavalue between 0 and 5 mF a range that shouldcover mo snsystemr inure todayfisheerfect oneosolutiuu is no greater than . Uontvqunnfiythe htterniftkod itrobusttocDunges in VD. Com scUDsinluleuoH tt especiPlu osefuifur . 1 gfadirns nnnhiithtiivt are insensitivctrfypical ckengetui Vofrom systemtosDftem SeriiooD .a . Excefsiaeseneikvityth cionnrwiamobiio-uPase p H is a crnnmoi reaoonfor a lack of mtthodenUottnertiaf . tfcomauter 51 11103 is used to s ptimiromoUile-phate pHc it ant alei determine ihe roODSfness oOfhe method with opsptct tvcma loariatiDusin fH. The tsmperatueeaí cnHPLC eotfemcan 0P coattolledir ucrimuswaysiSecuign but small 6. in wmeccumv earn uccuv basedon the vulec e U5 S. Again comeutce eimuivtioncan hr e vl io 6 tDu elleetof a ckavoein temperature onsrDacotion. Fortke lili motkod abtve Dig. lO-ket u iliansc 0u temperatureofU0 C resolie in tiers iu uetDiutiunkf no more tkan . Consequently tkirmetCoduad toierate .
