Method Development for Comprehensive Two-Dimensional Liquid Chromatography Separations

Method development continues to be a significant barrier to wider adoption of comprehensive two-dimensional liquid chromatography (LC×LC). Several groups have discussed different frameworks for systematically approaching method development over the past 15 years or so. In this chapter we propose a generic approach to developing LC×LC methods that leverages these past experiences and knowledge. The specific route one takes through this workflow will depend on which method performance metrics are most important for the application at hand. Then, we use two case studies of the development of real LC×LC methods to illustrate how the method development approach can be used in practice. Finally, we discuss strategies that can be used to optimize LC×LC methods, including recent progress toward automating this work. Method development for LC×LC is a highly active research area where we expect great strides to be made in the near future.