Recent progress of on-line sample preconcentration techniques in microchip electrophoresis

This review highlights recent developments and applications of on-line sample preconcentration techniques to enhance the detection sensitivity in microchip electrophoresis (MCE); references are mainly from 2008 and later. Among various developed techniques, we focus on the sample preconcentration based on the changes in the migration velocity of analytes in two or three discontinuous solutions system, since they can provide the sensitivity enhancement with relatively easy experimental procedures and short analysis times. The characteristic features of the on-line sample preconcentration applied to microchip electrophoresis (MCE) are presented, categorized on the basis of "field strength-" or "chemically" induced changes in the migration velocity. The preconcentration techniques utilizing field strength-induced changes in the velocity include field-amplified sample stacking, isotachophoresis and transient-isotachophoresis, whereas those based on chemically induced changes in the velocity are sweeping, transient-trapping and dynamic pH junction.     

Micellar electrokinetic chromatography: current developments and future

This review highlights recent methodological and instrumental advances in micellar electrokinetic chromatography (MEKC). Enhancements in sensitivity and selectivity of the technique through the use of on-line preconcentration approaches (stacking and sweeping) and nonconventional pseudostationary phases, namely nonionic and zwitterionic surfactants, mixed micelles and polymers, are discussed in detail. Laser-induced fluorescence and mass spectrometry, as alternatives to UV-absorption detection, have been covered to evaluate their advantages and limitations when applied to analysis in an MEKC format. Some thoughts on future directions in this area such as in-capillary reactions, coated capillaries and MEKC on microchips are also presented.     

Capillary electrophoresis on microchip

Capillary electrophoresis and related techniques on microchips have made great strides in recent years. This review concentrates on progress in capillary zone electrophoresis, but also covers other capillary techniques such as isoelectric focusing, isotachophoresis, free flow electrophoresis, and micellar electrokinetic chromatography. The material and technologies used to prepare microchips, microchip designs, channel geometries, sample manipulation and derivatization, detection, and applications of capillary electrophoresis to microchips are discussed. The progress in separation of nucleic acids and proteins is particularly emphasized.     

芯片毛细管电泳及其在生命科学中的应用

芯片毛细管电泳 (Chip CE)技术在近几年已取得了很大的进展。本文着重介绍芯片毛细管区带电泳技术 ,对等电聚焦、等速电泳、自由溶液电泳及胶束电动色谱等其它芯片电泳模式也有所提及。讨论了芯片材料和制作技术、芯片的几何形状、样品的操作和衍生、检测及芯片毛细管电泳技术的应用 ,特别是在核酸和蛋白质的分离分析中的进展     

Hydrophobic labeling of amino acids: transient trapping-capillary/microchip electrophoresis

Transient trapping (tr-trapping) was developed as one of the on-line sample preconcentration techniques to improve a low concentration-sensitivity in microchip electrophoresis (MCE), providing highly effective preconcentration and separation based on the trap-and-release mechanism. However, a poor performance to hydrophilic analytes limited the applicability of tr-trapping. To overcome this drawback, tr-trapping was combined with a sample labeling using a hydrophobic reagent in CE. Three commercially available fluorescent dyes, fluorescein isothiocyanate, succinimidyl esters of Alexa Fluor 488 and BODIPY FL-X, were tested as derivatization reagents to increase the hydrophobicity of amino acids (AAs) that were undetectable due to no fluorescence/UV-absorbance. As a result, it was confirmed that BODIPY labeling allowed various AAs to be analyzed in tr-trapping-micellar electrokinetic chromatography (tr-trapping-MEKC) by the increase in the hydrophobicity. In tr-trapping-MEKC, both the improvement of the resolution and 106-125-fold enhancements of the detectability of labeled AAs were achieved relative to the conventional capillary zone electrophoresis. The limit of detection of labeled phenylalanine was improved from 800 to 5 pM by applying tr-trapping-MEKC. In tr-trapping-microchip MEKC, furthermore, an 80-160-fold enhancement of the peak intensity and a baseline separation was also achieved within 30 s. These results clearly demonstrate that the tr-trapping technique with hydrophobic labeling will make CE/MCE more sensitive for various analytes.     

On-chip micellar electrokinetic chromatographic separation of phenolic chemicals in waters

This paper describes on-chip micellar electrokinetic chromatography (MEKC) separation of bisphenol A and 3 kinds of alkylphenols, which have been recently recognized as endocrine disrupting chemicals for fish by the Japanese government, using microchip capillary electrophoresis with UV detection. We successfully obtained high-speed separation of the phenolic chemicals within 15 s as optimizing in microfluidic controls and MEKC separation conditions. We obtained fairly good linearity with correlation coefficient of over 0.98 from 0 to 50 mg/l phenolic chemicals except for 4-nonylphenol, which sample is the mixture of many geometrical isomers (r = 0.86). The values of the relative standard deviation for peak height in 50 mg/l phenolic chemicals were less than 8% except for bisphenol A (11.0%). The limits of detection obtained at a signal-to-noise ratio of 3 were from 5.6 to 20.0 mg/l. To realize on-site monitoring, we described strategy for on-chip MEKC analysis of the phenolic chemicals in waters using a portable analyzer based on microfluidic devices.     

Poly(dimethylsiloxane) microchip for precolumn reaction and micellar electrokinetic chromatography of biogenic amines

We have demonstrated that precolumn derivatization and capillary electrophoresis separation on a poly(dimethylsiloxane) (PDMS) microchip can be realized as efficient as those on glass microchips. In an optimized condition of micellar electrokinetic chromatography (MEKC), using 25 mM sodium borate buffer (pH 10.0) with 25 mM sodium dodecyl sulfate (SDS) and 5% v/v methanol, the electroosmotic flow in an oxidized PDMS microchip is stabilized within 3% for days. By employing a fluorometric derivatization with o-phthaldialdehyde (OPA) in an optimally designed reaction chamber, four most important biogenic amines occurring in foods, histamine, tyramine, putrescine, and tryptamine, are quantitatively determined in less than 1 min at the levels applicable to real samples. The migration behaviors of anionic OPA-derivatized biogenic amines under the MEKC conditions are analyzed, and it has been found that under our separation conditions, the electrophoretic mobility of the SDS micelles is significantly greater than those of the anions in the aqueous phase. The channel manifold in a PDMS substrate is fabricated using replica molding against a thick photoresist, SU-8, pattern generated by photolithography. The plate with the microchannel pattern is strongly, irreversibly bonded to another PDMS plate by using a new bonding technique, which employs surface oxidation by corona discharge generated from a cheap, handy source, Tesla coil.     

High-throughput DNA analysis by microchip electrophoresis

DNA analysis plays a great role in genetic and medical research, and clinical diagnosis of inherited diseases and particular cancers. Development of new methods for high throughput DNA analysis is necessitated with incoming of post human genome era. A new powerful analytical technology, called microchip capillary electrophoresis (MCE), can be integrated with some experimental units and is characterized by high-speed, small sample and reagent requirements and high-throughput. This new technology, which has been applied successfully to the separation of DNA fragments, analysis of polymerase chain reaction (PCR) products, DNA sequencing, and mutation detection, for example, will become an attractive alternative to conventional methods such as slab gel electrophoresis, Southern blotting and Northern blotting for DNA analysis. This review is focused on some basic issues about DNA analysis by MCE, such as fabrication methods for microchips, detection system and separation schemes, and several key applications are summarized.     

Recent innovations in protein separation on microchips by electrophoretic methods

Microchips for analytical purposes have attracted great attention over the last 20 years. In the present review, we focus on the most recent development of microchips for electrophoretic separation of proteins. This review starts with a short recalling about the microchips covering the basic microchip layout for CE and the commercial chips and microchip platforms. A short paragraph is dedicated to the surface treatment of microchips, which is of paramount importance in protein analysis. One section is dedicated to on-line sample pretreatment in microchips and summarizes different strategies to pre-concentrate or to purify proteins from complex matrixes. Most of the common modes used for CE of proteins have already been adapted to the chip format, while multidimensional approaches are still in progress. The different routes to achieve detection in microchip are also presented with a special attention to derivatization or labeling of proteins. Finally, several recent applications are mentioned. They highlight the great potential of electrophoretic separations of proteins in numerous fields such as biological, pharmaceutical or agricultural and food analysis. A bibliography with 151 references is provided covering papers published from 2000 to the early 2007.     

Stacking and separation of enantiomers by acetonitrile-salt mixtures in micellar electrokinetic chromatography

The feasibility of employing the "acetonitrile stacking" method in micellar electrokinetic chromatography (MEKC) for the on-line preconcentration and separation of enantiomers is demonstrated for the first time. The effects of various experimental parameters on the stacking and separation of three different pairs of optical isomers, i.e., two substituted naphthyl enantiomers and one dansylated-DL-amino acid, were examined. In particular, the effectiveness of the addition of acetonitrile and salt in the sample matrix to induce narrowing of the analyte bands was investigated in the presence of sodium cholate as the chiral surfactant micelle in the separation buffer. For example, it was found that the presence of both acetonitrile and 1% NaCl in the sample matrix (volume ratio = 2:1) led to a significant improvement of the peak height and resolution for the MEKC separation of a pair of R(-)/S(+)-1,1'-binaphthyl diyl hydrogen phosphate enantiomers when the injection sample size was relatively large (e.g., 12% capillary volume). Furthermore, the feasibility of combining salting-out solvent extraction (off-line) and acetonitrile stacking (on-line) as a novel approach for sample preconcentration in capillary electrophoresis was also demonstrated.     

Electrophoretic analysis of food dyes using a miniaturized microfluidic system

A simple and sensitive on-chip preconcentration, separation, and electrochemical detection (ED) method for the electrophoretic analysis of food dyes was developed. The microchip comprised of three parallel channels: the first two are for the field-amplified sample stacking (FASS) and subsequent field-amplified sample injection (FASI) steps, while the third one is for the micellar EKC with ED (MEKC-ED) step. The food dyes were initially extracted from real samples by employing a method that was simpler, easier, and faster compared with a standard method. The extraction of the samples was characterized by UV-Vis and electrochemical experiments. The chronoamperometric detection was performed with a glassy carbon electrode coupled horizontally with the microchip at the separation channel exit. Experimental parameters affecting the analytical performance of the method were assessed and optimized. The sensitivity of the method was improved by approximately 10,800-fold when compared with a conventional MEKC-ED analysis. Reproducible response was observed during multiple injections of samples with an RSD of <7.2% (n=5). The calibration plots were linear (r2=0.998) within the range of 1.0 nM-1.0 microM for all food dyes. LODs were estimated between 1.0 and 5.0 nM, based on S/N=3, for food dyes. The applicability of the method for the analysis of food dyes in real sample was demonstrated.     

Recent progress of online sample preconcentration techniques in microchip electrophoresis

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.     

Recent developments in electrokinetically driven analysis on microfabricated devices

This review is devoted to the rapid developments in the field of microfluidic separation devices in which the flow is electrokinetically driven, and where the separation element forms the heart of the system, in order to give an overview of the trends of the last three years. Examples of microchip layouts that were designed for various application areas are given. Optimization of mixing and injection strategies, designs for the handling of multiple samples, and capillary array systems show the enormous progress made since the first proof-of-concept papers about lab-on-a-chip devices. Examples of functional elements for on-chip preconcentration, filtering, DNA amplification and on-chip detection indicate that the real integration of various analytical tasks on a single microchip is coming into reach. The use of materials other than glass, such as poly(dimethylsiloxane) and polymethylmethacrylate, for chip fabrication and detection methods other than laser-induced fluorescence (LIF) detection, such as mass spectrometry and electrochemical detection, are described. Furthermore, it can be observed that the separation modes known from capillary electrophoresis (CE) in fused-silica capillaries can be easily transferred to the microchip platform. The review concludes with an overview of applications of microchip CE and with a brief outlook.     

胶束毛细管电动色谱法在线富集联用技术灵敏检测乳制品和饲料样品中的三聚氰胺

建立了胶束毛细管电泳(MEKC)在线富集技术灵敏检测三聚氰胺的方法,采用场放大进样(FASS)联用胶束扫集(Sweep)测定多种样品中的三聚氰胺.试样用乙腈反复提取3次,在优化实验条件下,三聚氰胺的检测灵敏度提高了约1000倍,检出限由原来的2 mg/L降到1.8 μg/L(S/N=3).本方法用于配方奶粉和动物饲料中...     

Capillary electrophoresis in pharmaceutical analysis: a survey on recent applications

Capillary electrophoresis (CE) has a significant role in drug discovery and manufacturing processes and has a potential to grow further, due to new developments that can provide highly sensitive and high throughput analysis. This review illustrates recent applications of CE in pharmaceutical analysis (2005-present). The history, principles, instruments, and conventional modes of CE are briefly described. Applications for drug analysis by various techniques of CE are presented in six tables: capillary zone electrophoresis (CZE) (Table I), micellar electrokinetic chromatography (MEKC) and microemulsion electrokinetic chromatography (MEEKC) (Table II), non-aqueous CE (NACE) (Table III), chiral CE (Table IV), CE-mass spectrometry (MS) microchip CE (Table V), and multiplexed CE (MCE) (Table VI).     

Recent developments in electrochemical detection for microchip capillary electrophoresis

Significant progress in the development of miniaturized microfluidic systems has occurred since their inception over a decade ago. This is primarily due to the numerous advantages of microchip analysis, including the ability to analyze minute samples, speed of analysis, reduced cost and waste, and portability. This review focuses on recent developments in integrating electrochemical (EC) detection with microchip capillary electrophoresis (CE). These detection modes include amperometry, conductimetry, and potentiometry. EC detection is ideal for use with microchip CE systems because it can be easily miniaturized with no diminution in analytical performance. Advances in microchip format, electrode material and design, decoupling of the detector from the separation field, and integration of sample preparation, separation, and detection on-chip are discussed. Microchip CEEC applications for enzyme/immunoassays, clinical and environmental assays, as well as the detection of neurotransmitters are also described.     

On-line sample preconcentration in capillary electrophoresis. Fundamentals and applications

On-line preconcentration is one of the aspects of analytical method development using capillary electrophoretic techniques. The choice of the sample matrix alone can significantly alter both method sensitivity and separation efficiency. The recent trend to detect samples in narrower separation vessels also necessitates the need to improve detection sensitivity. The desire to detect very low levels of analytes using limited amounts of sample from biological specimens and the high separation efficiency obtainable using very large injections compared to classical small size injections also adds to this list. Indeed, one of the rich areas of research in the capillary electrophoresis field is on on-line sample preconcentration. More than 400 published research articles gathered from the http://www.webofscience.com from the year 2000 described a form of on-line preconcentration in capillary electrophoresis. This review provides a comprehensive table listing the applications of on-line preconcentration in capillary electrophoresis.     

阴离子耗尽进样胶束扫集毛细管电动色谱法在线富集测定甘草黄酮类化合物

建立了一个毛细管胶束电动色谱(MEKC)在线富集阴离子耗尽进样(ASEI)联用扫集(sweeping)技术测定3种甘草黄酮化合物(异甘草素、甘草素和甘草苷)的方法。考察了MEKC的分离条件和富集体系的优化条件,其中样品基质、水塞进入时间、进样时间对目标化合物的富集效果有较大的影响。在优化实验条件下,异甘草素、甘草素和甘草苷的富集倍数分别提高了110、120、300倍,检出限分别为0.015、0.014、0.011mg/L。该方法用于中药制剂中甘草素、异甘草素和甘草苷含量的测定,回收率在90.6%～107%之间,相对标准偏差(RSD)均小于4.5%(n=3)。实验证明,该方法可成功地应用于实际样品中甘草素、异甘草素和甘草苷含量的测定     

Separation of biogenic amines by micellar electrokinetic chromatography with on-line chemiluminescence detection

A method of on-line chemiluminescence detection with capillary electrophoresis for biogenic amines (diaminopropane, putrescine, cadaverine and diaminohexane) labeled with N-(4-aminobutyl)-N-ethylisoluminol is reported for the first time. Two separation modes, capillary zone electrophoresis and micellar electrokinetic chromatography (MEKC), were studied. The results show that excellent resolution was achieved in MEKC. Parameters affecting separation process and chemiluminescence detection have been examined in detail. Under the optimum conditions, the baseline separation of four amines was obtained within 7.5 min. The detection limits (S/N=3) of diaminopropane, putrescine, cadaverine and diaminohexane are 3.5 x 10(-8), 3.5 x 10(-8), 3.9 x 10(-8) and 1.2 x 10(-7) M, respectively. The method was applied to the analysis of biogenic amines in lake water.