The Practical Aspects of Normal-Phase HPLC

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Fang Xia, Diane M. Diehl, Jeffrey M. Mazzeo [Waters]
PREP 2006, May 14-17, Baltimore MD
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In today’s separation technology, reversed-phase liquid chromatography (RPLC) is the most widely used methodology due to its excellent resolving power and superior data reproducibility. However, normal-phase liquid chromatography (NPLC), as the first HPLC separation mode, is still very popular because of its unique selectivity, wide solvent choice, low system backpressure and easy solvent evaporation. A new normal-phase media made from high purity silica with maximized surface area has been developed. Several applications are presented that address the unique selectivity of NPLC for isomers, steroids, and classical synthetic batch reactants in medicinal chemistry labs. The preparative NPLC columns are manufactured with Optimum Bed Density (OBD™) design, which ensures the direct scale-up from analytical to preparative columns. The OBD™ preparative columns also possess the advantages of higher mass loading and longer column lifetime. The impact of several commonly used solvents to NPLC separations is also studied from a practical view point. In addition, the equilibrium of different pore size silica normal phase particles in the presence of water is compared for a better understanding of particle design and use.

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