Strategies for systematic HPLC method development

ANYL 244

Thomas Jupille, tjupille@sepsci.com and John Dolan. LC Resources Inc, 1295-F Boulevard Way, Walnut Creek, CA 94595
The objective of HPLC method development is the "identification of chromatographic conditions which meet the goals of the analysis". In an industrial setting, this objective must be modified to include the phrase "as quickly and as efficiently as possible".

Emphasizing reversed-phase chromatography, this presentation will explore the parameters (retention, selectivity, and efficiency) and the variables (column dimensions, particle size, flow rate, temperature, mobile phase strength, organic solvent type, column type, pH, and additive type/concentration) which must be manipulated in order to attain this objective. We will focus on identifying and ranking the key variables and outline strategies for systematically exploring their effects which maximizes the probability of meeting analytical goals while minimizing experimental time and effort.