Combination of on-line sample preconcentration by large-volume dual preconcentration by isotachophoresis and stacking (LDIS) with field-amplified sample injection (FASI) on Y-channel microchips.

In our previous study, the combination of two on-line sample preconcentration techniques, large-volume sample stacking with an electroosmotic flow (EOF) pump (LVSEP) and transient isotachophoresis (tITP), in microchip electrophoresis (MCE) was developed, which was named large-volume dual preconcentration by isotachophoresis and stacking (LDIS). LDIS was apparently effective for improving the sensitivity and the peak shape. In LDIS, however, there was a limit to the improvement of the sensitivity enhancement factor (SEF) since the amount of analytes to be concentrated was limited to the channel volume. To overcome this issue, in the present article, LDIS was coupled with field-amplified sample injection (FASI) technique on Y-shaped channel microchips. The use of a Y-channel in LDIS-FASI allowed consecutive LVSEP, FASI and tITP enrichments with a simple voltage control. In conventional LVSEP and LDIS analyses of a standard analyte, the SEFs were evaluated to be 2630 and 13,100, respectively, whereas in LDIS-FASI that was increased to 27,900 even at the FASI injection time of 0 s. To achieve higher SEFs, furthermore, the FASI injection time was increased to 150 s, resulting in the best SEF of 58,500. It should be emphasized that the peak width in LDIS-FASI was quite narrow, only 0.3-3.1 s, while in normal LVSEP that was 13 s. Furthermore, the LDIS-FASI technique was applied to the analysis of oligosaccharide mixture. Due to the focusing effect by LDIS-FASI, the resolutions were improved from 0.97-1.57 to 2.08-2.73.