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. Author manuscript; available in PMC: 2010 Jul 15.
Published in final edited form as: Electrophoresis. 2008 Apr;29(8):1604–1611. doi: 10.1002/elps.200700741

Figure 1.

Figure 1

Diagram of the advanced on-line 2D SCX/PLOT/MS system using a 3.2 m × 10 μm i.d. PLOT column and an on-line triphasic trapping column. For the latter column, a sol-gel frit was first made in 75 μm i.d. fused-silica tubing, followed by 2 cm of SCX resin (Polysulfoethyl A™) and then 2 cm of C18 RP packing. A 4 cm × 50 μm i.d. PS-DVB monolithic column was connected to the biphasic RP/SCX column to assemble the triphasic RP/SCX/RP trapping column. A PicoClear tee was used to minimize the dead volume between the triphasic trapping column and the PLOT column, while allowing fast sample loading. Microtees a and b were employed to split the mobile phase during SCX fractionation and PLOT separation, respectively. The sample was first injected onto the C18 column for desalting, then gradient eluted and trapped on the SCX packing, and finally stepwise eluted and trapped onto the monolithic micro-SPE column. The trapped sample was switched on-line to the PLOT column for separation. See text for the details.