In this lecture the advantages of comprehensive two-dimensional liquid chromatography (LC×LC) will be discussed. LC×LC may provide high peak capacities in a much shorter time than conventional 1D-LC. Moreover, LC×LC provides unique selectivity. It is an attractive option for separating the very complex mixtures that are encountered in many fields (life science, food science, material science, etc.). LC×LC is instrumentally straightforward, but the development and optimization of methods may be an obstacle. If method development in LC is a time-consuming task, then method development in LC×LC threatens to be a (time)2 consuming task. Some guidelines will be discussed to help understand LC×LC and to develop methods more efficiently.If we are able to “modulate” collected fractions from the first-dimension separation we may minimize the effects of the first-dimension eluent on the second-dimension separation and we may essentially optimize the two separations independently. This is demonstrated for the separation of peptides by a combination of ion-exchange chromatography and reversed-phase LC. After the first-dimension separation the analytes are focussed on a trapping column. This allows the second-dimension column to me much narrower than the first-dimension column and it provides an increase in sensitivity by several orders of magnitude. In addition, salt is removed from the eluent, which promotes the MS compatibility of the system. Finally, directions for further development of multi-dimensional LC methods will be discussed.
Presenting author:
Peter Schoenmakers
University of Amsterdam, Faculty of Science, Science Park 904, 1098 XH Amsterdam, The Netherlands
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Authors:
Peter Schoenmakers - University of Amsterdam
