Making the Most of Your HPLC Separations: Maximizing Selectivity, Peak Capacity and Speed

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Diane M. Diehl, Eric S. Grumbach, Jeffrey R. Mazzeo, Uwe D. Neue [Waters]
ACS Spring Meeting
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Developing HPLC methods is often a time-consuming and daunting task. Many chromatographers have their own unique approach to method development. Some try a particular column at one pH, while others use a bank of columns with a variety of mobile phases. No universal approach to method development exists. To address the needs of current chromatographers for a more systematic approach to method development, we have examined how pH, stationary phase and solvent choice influence the selectivity of a separation. By far, the mobile phase pH has the most influence on selectivity. From these data, we have developed an approach to selecting the column and mobile phase for a particular separation. As chromatographers move towards faster method development, we discuss how peak capacity can be used to measure the separation power of a column. Combining the factors that control selectivity with peak capacity measurements, we can develop robust methods faster. Additionally, new advances in HPLC instrumentation and column technology that enable faster method development will be discussed.