Abstract
Method development can be straightforward for simple samples, where target, degradant and sample matrix compounds are known, and critical peak pairs resolved. For complex samples, the sample’s own matrix complicates the separation, with many unknown analytes that can co-elute and lead to a loss of resolution between critical peak pairs. Therefore, the best approach for complex sample separations is to maximize the separation power afforded by the accessible separation and detection technologies and subsequent optimization of the method to gain the highest practical peak capacity.
In this talk we highlight a practical approach, based on a thorough fundamental and practical understanding of reversed phase LC (RPLC) gradient elution theory. A guide that is accessible to all analysts and exploits the sample to guide the method development and column selection decisions. Two protocols will be shared, that are well established and used in practice for complex samples encountered across different industries and applications. One protocol is aimed at relatively larger peptide type separations and another, focused on small molecules.