This paper describes a novel on-line sample preconcentration and separation technique named transient trapping (tr-trapping), which improves the efficiencies of separation and concentration by using a partially injected short micellar plug in microchip electrophoresis. Although a longer separation length often provides a better resolution of complexed or closely migrating analytes, our proposed theoretical model indicated that a trap-and-release mechanism enables a short micellar zone, which was partially injected into the separation channel, to work as an effective concentration and separation field. Application of the tr-trapping technique to microchip micellar electrokinetic chromatography (MCMEKC) was performed on a newly fabricated 5-way-cross microchip by using sodium dodecyl sulfate and rhodamine dyes as test micelle and analytes, respectively. When the injection times of micelle (t(inj),M) and sample solution (t(inj),S) were 1.0 and 2.0 s, respectively, both the preconcentration and separation of the dyes were completely finished within only 3.0 s. At t(inj),S of 8.0 s, a 393-fold improvement of the detectability was achieved in comparison with conventional MCMEKC. The resolution obtained with tr-trapping-MCMEKC was also better than that with conventional MCMEKC in spite of the 160-fold shorter length of the injected micellar zone at t(inj),M of 1.0 s. These results clearly demonstrated that the tr-trapping technique in MCMEKC provides a rapid, high-resolution and detectability analysis even in the short separation channel on the microchips.

The applicability of an online preconcentration technique, large-volume sample stacking with an electroosmotic flow pump (LVSEP), to microchip zone electrophoresis (MCZE) for the analysis of oligosaccharides was investigated. Since the sample stacking and separation proceeded continuously without polarity switching in LVSEP, a single "straight" channel microchip could be employed. In the MCZE analysis of oligosaccharides, sample adsorption onto the channel surface should be suppressed, so the straight microchannel was modified with poly(vinyl alcohol) (PVA). So far, the mechanism of LVSEP in the polymer-coated capillary or microchannel has not been reported, and thus, the LVSEP process in the PVA-coated channel was investigated by fluorescence imaging. Although it is well-known that the PVA coating can suppress the electroosmotic flow (EOF), an enhanced EOF with a mobility of 4.4 x 10(-4) cm(2)/(V x s) was observed in a low ionic strength sample solution. It was revealed that such temporarily enhanced EOF in the sample zone worked as the driving force to remove the sample matrix in LVSEP. To evaluate the analytical performance of LVSEP-MCZE, oligosaccharides were analyzed in the PVA-coated straight channel. As a result, both the glucose ladder and oligosaccharides obtained from bovine ribonuclease B were well enriched and separated with up to 2200-2900-fold sensitivity enhancement compared to those in a conventional MCZE analysis. The run-to-run repeatabilities of the migration time and peak height were good with relative standard deviations of 1.1% and 7.2%, respectively, which were better than those of normal MCZE. By applying the LVSEP technique to MCZE, a complicated voltage program for fluidic control could be simplified from four channels for two steps to two channels for one step.

Microchip electrophoresis (MCE) has been advanced remarkably by the applications of several separation modes and the integration with several chemical operations on a single planer substrate. MCE shows superior analytical performance, e.g., high-speed analysis, high resolution, low consumption of reagents, and so on, whereas low-concentration sensitivity is still one of the major problems. To overcome this drawback, various online sample preconcentration techniques have been developed in MCE over the past 15 years, which have successfully enhanced the detection sensitivity in MCE. This review highlights recent developments in online sample preconcentration in MCE categorized on the basis of "dynamic" and "static" methods. The dynamic techniques including field amplified stacking, ITP, sweeping, and focusing have been easily applied to MCE, which provide effective enrichments of various analytes. The static techniques such as SPE and filtration have also been combined with MCE. In the static techniques, extremely high preconcentration efficiency can be obtained, compared to the dynamic methods. This review provides comprehensive tables listing the applications and sensitivity enhancement factors of these preconcentration techniques employed in MCE.

The refractive index sensitivity of a localized surface plasmon resonance (LSPR) sensor is correlated to an enhanced local electromagnetic (EM) field originating from noble metal nanostructures. Here, we demonstrated that extensive EM field enhancement by a gold (Au) nanocone array (AuNCA) allowed highly sensitive and label-free detection of biomolecules in the visible wavelength spectrum. The AuNCA consisted of a polymer core and an Au shell, which was fabricated by using simple and inexpensive nanoimprint lithography. Under LSPR excitation, AuNCA absorbs visible light of a specific wavelength and extensively enhances the EM field near its surface. It was shown that AuNCA had high refractive index sensitivity (417.5 nm/RIU) because of the large distribution of the enhanced EM field, covering a large surface of the NCA. Moreover, in DNA hybridization detection, a very low limit of detection of 161 fM was achieved, and 1-base mismatch DNA was successfully discriminated by using AuNCA.

To improve the sensitivity in chiral analysis by capillary electrophoresis without loss of optical resolution, application of large-volume sample stacking with an electroosmotic flow pump (LVSEP) was investigated. Effects of the addition of cyclodextrin (CD) into a running solution on the LVSEP preconcentration was theoretically studied, where the preconcentration efficiency and effective separation length would be slightly increased if the effective electrophoretic velocity (v(ep,eff,BGS)) of the analytes was decreased by interacting with CD. In LVSEP-CD-modified capillary zone electrophoresis (CDCZE) and LVSEP-CD electrokinetic chromatography with reduced v(ep,eff,BGS), up to 1000-fold sensitivity increases were achieved with almost no loss of resolution. In LVSEP-CD-modified micellar electrokinetic chromatography of amino acids with increased v(ep,eff,BGS), a 1300-fold sensitivity increase was achieved without much loss of resolution, indicating the versatile applicability of LVSEP to many separation modes. An enantio-excess (EE) assay was also carried out in LVSEP-CDCZE, resulting in successful analyses of up to 99.6% EE. Finally, we analyzed ibuprofen in urine by desalting with a C₁₈ solid-phase extraction column. As a typical result, 250ppb ibuprofen was well concentrated and optically resolved with 84.0-86.6% recovery in LVSEP-CDCZE, indicating the applicability of LVSEP to real samples containing a large amount of unnecessary background salts.