The ever increasing demands on chromatographers to rapidly produce fit for purpose separations requires more efficient and methodical working processes. Method development workflows are generally well established for reversed phase liquid chromatography. Hydrophillic Interaction Liquid Chromatography (HILIC) is less well understood practically and mechanistically, meaning that the method development process tends to be less systematic and well defined. This ACE Knowledge Note details a simple, rationally designed protocol for HILIC method development using the three ACE HILIC phases.
Efficient method development procedures require a logical exploration of key chromatographic parameters leading to identification of a robust method on a suitable column/mobile phase combination. Rationally designed method development procedures assess key parameters which affect chromatographic selectivity (e.g. stationary phase, pH etc.) and enable analysts to make well informed decisions, whilst reducing the risk of developing sub standard, non robust methods. By following a step by step process, method development can be streamlined thereby increasing laboratory productivity.
The approach outlined here is based on a logical assessment of the most powerful parameters affecting HILIC selectivity.
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