Application of on-line sample preconcentration by large-volume dual preconcentration by isotachophoresis and stacking (LDIS) on straight-channel microchips

IF 1.8 4区 化学 Q3 CHEMISTRY, ANALYTICAL
Fumihiko Kitagawa, Kazuki Takahashi, Reina Osanai, Ryota Sasaki, Takayuki Kawai
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Abstract

In this study, large-volume dual preconcentration by isotachophoresis and stacking (LDIS) which is an on-line sample preconcentration technique coupling large-volume sample stacking with an electroosmotic flow pump (LVSEP) with transient isotachophoresis (tITP) was applied to microchip electrophoresis (MCE) for improving both detection sensitivities and peak shapes. To realize LDIS in MCE, we investigated experimental procedures for injecting a short plug of a leading electrolyte (LE) solution into a straight microchannel without any sophisticated injector apparatus. We found that a short LE plug could be injected into a sample-filled straight-channel only by making the liquid level of the LE solution in an outlet reservoir higher than that in an inlet one. By applying a reversed-polarity voltage to the microchip, anionic analytes injected throughout the microchannel were first enriched by LVSEP, followed by tITP. Through the second preconcentration effect by tITP in LDIS, sensitivity enhancement factor (SEF) and asymmetry factor for a standard dye were improved from 878 and 0.62 to 1330 and 1.14, respectively, relative to those in conventional LVSEP. It should be noted that more viscous running buffer containing sieving polymers could be employed to the LDIS analysis, which was effective for improving the SEF and the separation efficiencies, especially for bio-polymeric compounds. Finally, LDIS was applied to the oligosaccharide and protein analyses in MCE, resulting in the SEFs of 1410 and ca. 50 for maltotriose and bovine milk casein, respectively.

Graphical Abstract

Abstract Image

在直通道微芯片上应用大容量同向电泳和堆积双预浓缩(LDIS)在线样品预浓缩技术。
在这项研究中,我们将大容量等渗和堆积双重预浓缩技术(LDIS)应用于微芯片电泳(MCE),该技术是一种在线样品预浓缩技术,它将大容量样品堆积与电渗流泵(LVSEP)和瞬态等渗电泳(tITP)结合在一起,从而提高了检测灵敏度和峰形。为了在 MCE 中实现 LDIS,我们研究了在不使用任何复杂的注入装置的情况下,将前导电解质(LE)溶液的短塞注入直的微通道的实验程序。我们发现,只有使出口储液器中的前导电解质溶液液面高于进口储液器中的液面,才能将前导电解质短塞注入充满样品的直通微通道。通过在微芯片上施加反极性电压,注入整个微通道的阴离子分析物首先被 LVSEP 富集,然后被 tITP 富集。通过 LDIS 中 tITP 的二次预富集效应,标准染料的灵敏度增强因子(SEF)和不对称因子分别从 878 和 0.62 提高到 1330 和 1.14,而传统 LVSEP 的灵敏度增强因子和不对称因子则分别从 878 和 0.62 提高到 1330 和 1.14。值得注意的是,在 LDIS 分析中可以使用含有筛分聚合物的更粘稠的运行缓冲液,这对提高 SEF 和分离效率非常有效,尤其是对生物聚合物化合物。最后,将 LDIS 应用于 MCE 中的寡糖和蛋白质分析,结果麦芽三糖和牛乳酪蛋白的 SEF 分别为 1410 和约 50。
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来源期刊
Analytical Sciences
Analytical Sciences 化学-分析化学
CiteScore
2.90
自引率
18.80%
发文量
232
审稿时长
1 months
期刊介绍: Analytical Sciences is an international journal published monthly by The Japan Society for Analytical Chemistry. The journal publishes papers on all aspects of the theory and practice of analytical sciences, including fundamental and applied, inorganic and organic, wet chemical and instrumental methods. This publication is supported in part by the Grant-in-Aid for Publication of Scientific Research Result of the Japanese Ministry of Education, Culture, Sports, Science and Technology.
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