Two-Dimensional HPLC Using Polymer Monoliths

Polymer monolithic columns with small diameter have been successfully employed for proteome analysis. Karger and coworkers reported MALDI-TOF of separated fractions spotted on a plate from a polymeric reversed-phase column that showed high peak capacity (Chen et al., 2005). Huber and coworkers reported separation and detection of about 200 peaks within 5 min by using a PSDVB column (Premstaller et al., 2001). 

As mentioned earlier, high-speed separation is necessary to carry out fast, comprehensive 2D HPLC. The polymer monoliths have not been employed in such 2D HPLC, probably because permeability of polymer monoliths is not high enough to allow fast elution of the second dimension (2nd-D) in simple 2D operation, and the gradient cycle at the 2nd-D cannot be so fast to allow online 2D operation without reducing peak capacity at first dimension (lst-D). 

The application of polymer monoliths in 2D separations, however, is very attractive in that polymer-based packing materials can provide a high performance, chemically stable stationary phase, and better recovery of biological molecules, namely proteins and peptides, even in comparison with C18 phases on silica particles with wide mesopores (Tanaka et al., 1990). Microchip fabrication for 2D HPLC has been disclosed in a recent patent, based on polymer monoliths (Corso et al., 2003). This separation system consists of stacked separation blocks, namely, the first block for ion exchange (strong cation exchange) and the second block for reversed-phase separation. This layered separation chip device also contains an electrospray interface microfabricated on chip (a polymer monolith/ 

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