Approaches to comprehensive multidimensional liquid chromatography systems

This work compares the performance of the three different schemes implementing two-dimensional liquid chromatography (2D-LC) in terms of the peak capacity that they can generate and of the time that they need to complete a two-dimensional analysis. We discuss in detail how time is spent in these two-dimensional liquid chromatography×liquidchromatography×liquid chromatography (LC×LCLC×LC) schemes and how to compare them. Keeping constant the characteristics of the first-dimension separation, we systematically varied those of the second-dimension separation and of its coupling to the first-dimension. In the process, five systems were created, based on the principles of the three known implementations of comprehensive 2D-LC. This work demonstrates an original method for the selection of the best comprehensive 2D-LC approach, depending on the desired peak capacity and on time constraints. The decision to use a 2D-LC method arises from the need to achieve a given resolution (i.e., a target peak capacity) within as short a time as possible or to reach the highest possible resolution in a given analysis time. Using the most appropriate schemes, we suggest how it is realistically possible to generate peak capacities ranging from 266 in just over 20 min or about 2800 in 2.3 h. When the time available for a two-dimensional separation is very short and the desired peak capacity cannot be achieved in 1D-LC, an on-line 2D-LC approach is unquestionably best. However, if a longer analysis time is acceptable, a 10-fold increase in the peak capacity can be obtained at the cost of a mere 7-fold increase in total analysis time.