TAILIEUCHUNG - Introduction to Modern Liquid Chromatography, Third Edition part 52

Introduction to Modern Liquid Chromatography, Third Edition part 52. 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. . | 466 GRADIENT ELUTION Table Dependence of k on B for Different Separation Modes Eq. or Separation Mode Dependence of k on B Definition of kw Eq. or k Eq. Dependence of 5 or n on the solute Reversed-phase RPC Eq. value of isocratic k for A-solvent itvarcr or buffer as mobile phase S increases with solute molecular size Eq. Normal-phase NPC Chapter 7 Eq. vaiue of isocratic k ior B-solvent moreprlar solvent as mobile phase a increases wifhthc number 0f polae groups in the solute mr ecule Soctian Ion-exchange IPC Chapter 7t Eq. 4 5 value of isocratic k for inott phrse w sah concentration ss Iso M n mmiber orcharges or estate molecule .SUmeS ncai ilent buffer Hydrophobic interaction HIC Chapter 13O Eq. kw value of isocratic k for A-solvent as mobile phase higrs raef concentratiou S H Hydrophilic interaction HILIC Chapter 8 Eq. kB value ofiioooaScfefor A-solvent organic as mobile n increases widttOs number ot polar proqis 111 tlie sotaoe molecule Section Here the valueofw varies for different solutes andseparation modes as definedin Table f and f o refer to values of b at the beginning 0 and end a oa the gradient respectively. For small-molecule samples values of n for different separation modes usually vary between 1 and 4 a value of n 2 can be used as a starting approximation prior to meahodaevelepme nt .Thus for IhaOC sepaaotion with al 50 u column a flow rateof 2 mL min and gradiaoto fl 0 -10 h B inl0minutes o4e value of 4 woui4 beapptoximately 10 x 2 x 2 x log 10 that is not much different fsom the usluvoa 4 4 forsimilse RPC so IEC separationweeha 15Sx aolumn s flow raxaofa e nao andgroniaiitoS 10-100 mM in 10 same odlueof sosaltsfeemdauarion . note also for IES fdaothesalt ooncsntradon UHs aOr sumofc oocentrations of oak plur buffer . Foa further dltaiieoothtthsoldoSyadient elution for feparation modes other than RPC reo aOapeesUS cS l

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