Fast HPLC Separations: Theory and Practice

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Uwe D. Neue;Diane M. Diehl;Eric S. Grumbach;KimVan Tran;Jeffrey R. Mazzeo [Waters]
Pittcon 2003
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Anilines; Caffeine; Ketoprofen; Fenoprofen; Flurbiprofen; Ibuprofen; Epicatechin; Hydrocortisone; Perylene; Triamcinolone; Acetanilide; Sulfa drugs; Sulfonamides; Atenolol; Pindolol; Metoprolol; Steroids; Prednisolone; Cortisone; Nortestosterone; Estradiol; Testosterone; Corticosterone; Estrone; Progesterone
In today''s world, the analytical chemist is under a steady pressure to accomplish more analyses in less and less time. In HPLC, analysis time is determined by the separation power required, and is constrained by the pressure available. However, even within these limits, a significant optimization is possible, and the analysis time can be driven to under one minute (1,2) In this presentation, we will outline the principles of our approach (3), and show the tools that make rapid analyses possible. We will discuss the impact of the oprating conditions, and the influence of column length, particle size and column diameter. In the same context, we will also look at alternative column designs, such as monoliths. Most of our focus is the speed of the analysis, but we will also examine other important elements of an assay, such as sensitivity. Specifically, we will optimize the performance of 2 and 3 µm high-performance particles in short columns. The current work is an extension of previous studies (1,3) that examined the optimization of chromatographic speed from a very fundamental standpoint. At the same time, many practical examples of very-high-speed chromatography will be shown that can be accomplished with the tools that are available today.

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