Detailed comparison of in-house developed and commercially available heart-cutting and selective comprehensive two-dimensional liquid chromatography systems.

Recently, two-dimensional liquid chromatography (2D-LC) has become a popular approach to analyze complex samples. This is partly due to the introduction of commercial 2D-LC systems. In the past, 2D-LC was carried out on in-house developed setups, typically consisting of several switching valves and sample loops as the interface between the two dimensions. Commercial systems usually offer different 2D-LC modes in combination with specialized software to operate the instrument and analyze the data. This makes them highly user-friendly, however, at an increased cost compared to in-house developed setups. This study aims to make a comparison between an in-house developed 2D-LC setup and a commercially available 2D-LC instrument. The comparison is made based on experimental differences, in addition to more general differences, including cost price, flexibility, and ease of operation. Special attention is also paid to the different strategies to deal with the mobile phase incompatibility between the highly orthogonal separation mechanisms considered in this work: hydrophilic interaction liquid chromatography (HILIC) and reversed-phase LC (RPLC). For the commercial 2D-LC instrument, this is done using active solvent modulation (ASM), a valve-based approach allowing the on-line dilution of the effluent eluting from the first dimension column before transfer to the second dimension (D) column. For the in-house developed setup, a combination of restriction capillaries and a trap column is used. Using a sample of 28 compounds with a large polarity range, peak shapes and recoveries of the D-chromatograms are compared for both setups. For early eluting compounds, the selective comprehensive approach, currently only possible on the commercial 2D-LC instrument, results in the best peak shapes and recoveries, however, at the cost of an increased analysis time. In general, depending on the analytical goal (single heart-cut versus full-comprehensive 2D-LC), an in-house developed system can be satisfactory for the analysis of specific target compounds/samples. For more complex problems, it can be interesting to use a more specialized commercial 2D-LC instrument. Overall, this comparison study provides advice for analytical scientists, who are considering to use 2D-LC, on the type of equipment to consider, depending on the needs of their particular applications.