Constant pressure mode of operation in the second dimension of two-dimensional liquid chromatography: A proof of...

The use of two-dimensional liquid chromatography (2D-LC) continues to grow as the advantages over 1D-LC become increasingly clear in specific application areas, and the number of experienced 2D-LC users increases. As with any technique, however, there is always room for innovation that could improve the performance of 2D-LC. In recent years the technical aspects and potential benefits of a volume-based mode of operation were studied in detail for 1D-LC. The salient features of this approach that are immediately interesting for use in 2D-LC are two-fold. First, the ability to maintain a nominally constant pressure in the second dimension by dynamically adjusting the flow rate to compensate for changes in the viscosity of the fluid in the ²D flow path provides a means to more fully utilize the pressure capability of the pumping system, and accelerates separations in the second dimension (²D). Second, constant pressure operation minimizes physical stress on the system components and the ²D column. In this paper we discuss the aspects of volume-based operation of LC that are particularly relevant to 2D-LC systems. The proof-of-concept experiments illustrate the viability of the constant pressure mode of operation for the second dimension of 2D-LC. In the described separations the throughput improvement is on the order of 10%; this gain will be strongly application-dependent, and may be as large as several tens percent in some cases. Future work will involve a detailed investigation of the impact of the constant pressure mode on robustness of ²D separations.