Application of CE in hydrodynamically closed systems for analysis of bioactive compounds in food

CE is a family of electrokinetic separation techniques that separate compounds based upon differences in electrophoretic mobilities, phase partitioning, pI, molecular size, or a combination of one or several of these properties. CE has been used in several modes to analyze and characterize a wide variety of analytes from simple inorganic ions, small organic molecules, peptides, proteins, nucleic acids to virus, microbes and particles. Food consists of a complex mixture of a variety of components, many of which are biologically active. Components classified as "nutrients" are essential for growth, maintenance, and repair of the body. Other food constituents, typically occurring in small quantities, are classified as "biologically active substances" and they have beneficial or harmful effects on human health. There are two types of biologically active substances in food - naturally occurring and food additives. The bioactive compounds of food that will be mentioned in this review are inorganic and organic acids, amino acids, vitamins, phenolic compounds, biogenic amines, antinutrients, toxins, etc. This review is focused on the application of CE with hydrodynamically closed system (suppression of EOF) for the analysis of the above-mentioned compounds. CE can be an alternative method to HPLC or other methods for analysis of bioactive compounds in food. The main advantages of CE are low running cost (at least ten times than HPLC) and consideration to environment (hundreds of microliters of diluted water based electrolyte per analysis).     

Analysis of glycerophospho‐ and sphingolipids by CE

Glycerophospholipids are amphiphilic molecules possessing polar head groups with a glycerol backbone and nonpolar variable long‐chain fatty acids. Numerous molecular species are found in a single class of glycerophospholipid, conferring to these lipids a high structural diversity. They are major components of biological membranes and participate in important activities involving cell signaling and substrate transport. Sphingolipids consist of long‐chain bases linked by an amide bond to a fatty acid and via the terminal hydroxyl group to complex carbohydrate or phosphorus moieties, constituting a complex family of compounds that also present an enormous structural variability. As important component of neuronal membranes, sphingolipids contribute to cellular diversity and functions and are associated with several neurodegenerative disorders. Moreover, they were studied in several foods due to their sensorial, reological, and antioxidant characteristics. In this work, the most relevant information available on glycerophospholipid and sphingolipid analysis by CE is reviewed. CE is a very promising analytical technique in polar lipid analysis, which provides high efficiency, relatively high resolution, and enormous versatility and requires small amounts of sample and solvent. MEKC and NACE methodologies have been developed as the most useful alternatives for these analyses by CE. Very interesting LODs have been achieved enabling the application of CE to the determination of glycerophospholipids and sphingolipids in several food and biological matrices.     

Capillary Electrophoresis Coupled On-Line with Inductively Coupled Plasma Atomic Emission Spectrometry for Elemental Speciation

Capillary electrophoresis (CE) has become a powerful analytical technique for the separation of a variety of analytes ranging from small inorganic ions to large biomolecules such as proteins and nucleic acids. A selective and sensitive detector for CE has been one of the most important and challenging prerequisites for the growth of CE. On-column UV-Vis detectors are commonly used to determine the analytes separated by CE. However, these detectors are often not very selective. Other detection techniques such as mass spectrometry, laser induced fluorescence, amperometry, and inductively coupled plasma spectrometry have been investigated to provide a more sensitive and selective detection for the target analytes. However, relatively few studies have been published on the use of inductively coupled plasma atomic emission spectrometry (ICP-AES) as a means of detection in CE separation.     

Capillary electrophoresis mass spectrometry and its application to the analysis of biological mixtures

Capillary electrophoresis (CE) mass spectrometry (MS), with its ability to separate compounds present in extremely small volume samples rapidly, with high separation efficiency, and with compound identification capability based on molecular weight, is an extremely valuable analytical technique for the analysis of complex biological mixtures. The highest sensitivities and separation efficiencies are usually achieved by using narrow capillaries (5-50 micro m i.d.) and by using sheathless CE-to-MS interfaces. The difficulties in CE-to-MS interfacing and the limited loadability of these narrow columns, however, have prevented CE-MS from becoming a widely used analytical technique. To remedy these limitations, several CE-MS interfacing techniques have recently been introduced. While electrospray ionization is the most commonly used ionization technique for interfacing CE-to-MS, matrix assisted laser desorption ionization has also been used, using both on-line and off-line techniques. Moreover, the high concentration detection limit of CE has been addressed by development of several sample concentration and sample focusing methods. In addition, a wide variety of techniques such as capillary zone electrophoresis, capillary isoelectric focusing, and on-column transient isotachophoresis have now been interfaced to MS. These advances have resulted in a rapid increase in the use of CE-MS in the analysis of complex biological mixtures. CE-MS has now been successfully applied to the analysis of a wide variety of compounds including amino acids, protein digests, protein mixtures, single cells, oligonucleotides, and various small molecules relevant to the pharmaceutical industry.     

毛细管电泳技术在疾病预防控制领域的应用、发展与挑战

自1989年出现商品化的仪器以来,毛细管电泳（CE）技术在多个应用领域都取得了长足的进步与发展,重复性和准确性方面也有很大提升。能力验证样品分析的满意结果也显示了CE具备法规要求的准确定量能力。在疾病预防控制领域（简称"疾控"）CE也展现出很多独具特色的应用,成为不可或缺的技术之一。在聚合酶链式反应产物分析、核酸序列测定、DNA变异和分型分析、食源性致病微生物分析及疫苗分析等工作中CE发挥了重要作用。应对突发疫情或公共卫生事件如食物中毒时,除了通过非靶标分析尽快锁定目标物外,还需要对大量样品做出快速而准确的分析,高通量和高灵敏的CE就十分适合解决这一问题。在公共卫生理化检验以确保食品、保健食品、特殊医学用途食品、化妆品和消毒产品等的安全中,CE也发挥了不可或缺的作用。作为一种使用较少危险化学品的环境友好方法,在需要按照标准或规范进行的疾控实验室常规检测中,CE仍受制于标准方法的缺失,未能发挥其应有的作用。但简单、快速、经济、耐用、高效的CE分离一旦与高灵敏通用检测器联用,必将更加从容地应对疾控领域中的各种挑战,发挥更大的作用。本文综述了2010~2019年CE在疾控领域的应用,分析了CE在疾控领域发展的机遇和挑战,对CE在疾控领域的发展前景进行了展望。     

Capillary electrophoresis for the analysis of small-molecule pharmaceuticals

This paper reviews the application of CE to the analysis of small-molecule pharmaceuticals. The areas of pharmaceutical analysis covered are enantiomer separation, the analysis of small molecules such as amino acids or drug counter-ions, pharmaceutical assay, determination of related substances and physicochemical measurements such as log P and pK(a) of compounds. The different electrophoretic modes available and their advantages for pharmaceutical analysis are described. Recent applications of CE for each subject area are tabulated with electrolyte details.     

Contributions of capillary electrophoresis to neuroscience

Capillary electrophoresis (CE) is a small-volume separation approach amenable to the analysis of complex samples for their small molecule, peptide and protein content. A number of the features of CE make it a method of choice for addressing questions related to neurochemistry. The figures of merit inherent to CE that make it well suited for studying cell-to-cell and intracellular signaling include small sample volumes, high separation efficiency, the ability for online analyte concentration, and compatibility with sensitive and high-information content detection methods. A variety of instrumental aspects are detailed, including detection methods and sampling techniques that are particularly useful for the analysis of signaling molecules. Studies that have used these techniques to increase our understanding of neurobiology are emphasized throughout. One notable application is single neuron chemical analysis, a research area that has been greatly advanced by CE.     

Recent advances in the application of capillary electrophoresis for food analysis

This article reviews recent developments in the application of capillary electrophoresis (CE) for the analysis of foods and food components. CE has been applied to a number of important areas of food analysis and is fast becoming an established technique within food analytical and research laboratories. Papers are reviewed that were published during the two years to date following the previous review (Electrophoresis 2001, 22, 4197-4206).     

CE microchips: an opened gate to food analysis

CE microchips are the first generation of micrototal analysis systems (-TAS) emerging in the miniaturization scene of food analysis. CE microchips for food analysis are fabricated in both glass and polymer materials, such as PDMS and poly(methyl methacrylate) (PMMA), and use simple layouts of simple and double T crosses. Nowadays, the detection route preferred is electrochemical in both, amperometry and conductivity modes, using end-channel and contactless configurations, respectively. Food applications using CE microchips are now emerging since food samples present complex matrices, the selectivity being a very important challenge because the total integration of analytical steps into microchip format is very difficult. As a consequence, the first contributions that have recently appeared in the relevant literature are based primarily on fast separations of analytes of high food significance. These protocols are combined with different strategies to achieve selectivity using a suitable nonextensive sample preparation and/or strategically choosing detection routes. Polyphenolic compounds, amino acids, preservatives, and organic and inorganic ions have been studied using CE microchips. Thus, new and exciting future expectations arise in the domain of food analysis. However, several drawbacks could easily be found and assumed within the miniaturization map.     

Recent advances in capillary electrophoresis methods for food analysis

This review article addresses recent advances in the analysis of foods and food components by capillary electrophoresis (CE). CE has found application to a number of important areas of food analysis, including quantitative chemical analysis of food additives, biochemical analysis of protein composition, and others. The speed, resolution and simplicity of CE, combined with low operating costs, make the technique an attractive option for the development of improved methods of food analysis for the new millennium.     

Measuring D-amino acid-containing neuropeptides with capillary electrophoresis

Neuropeptides are heavily posttranslationally modified (PTM) gene products that are often characterized by a variety of mass spectrometric approaches. Recently, the occurrence of amino acids in the D-form has been documented in several neuropeptides. As this modification has no associated mass shift, this particular PTM is difficult to evaluate using mass spectrometry (MS) alone. Here we demonstrate several approaches using capillary electrophoresis (CE) with absorbance and laser-induced fluorescence (LIF) for the separation of native and derivatized molluscan peptides containing D-amino acids. The combination of peptide derivatization followed by CE/LIF is well suited for single cell measurements because of its ability to characterize the peptides in such small samples. In order to verify this approach, the D-Trp-containing peptide NdWFa (NH2-Asn-D-Trp-Phe-CONH2), present in individual neurons from the marine mollusk Aplysia californica, has been characterized. The mass spectra show that NdWFa and/or NWFa are present in specific neurons; CE/LIF analysis of these cells demonstrates that NdWFa is the dominant form of the peptide.     

Capillary electrophoresis of quantum dots: Minireview

It has been already three decades, since the fluorescent nanocrystals called quantum dots (QDs) appeared and attracted attention of a broad scientific community. Their excellent not only optical but also electronic properties predetermined QDs for utilization in a variety of areas. Besides lasers, solar cells, and/or computers, QDs have established themselves in the field of (bio)chemical labeling as well as medical imaging. However, due to the numerous application possibilities of QDs, there are high demands on their properties that need to be precisely controlled and characterized. CE with its versatile modes and possibilities of detection was found to be an effective tool not only for characterization of QDs size and/or surface properties but also for monitoring of their interactions with other molecules of interest. In this minireview, we are giving short insight in analysis of QDs by CE, and summarizing the advantages of this method for QDs characterization.     

Perspectives on analyses of nucleic acid constituents: the basis of genomics

The recent mapping of the human genome was a tremendous achievement made possible to a large degree by the development of analytical methods for sequencing purine and pyrimidine bases in nucleic acids. In the last 3 decades, the number of analyses of nucleic acids and their constituents by HPLC and capillary electrophoresis (CE) has exploded. These techniques have been used not only for genomics, but also for the determination of free nucleotides, nucleosides and their bases in body fluids and tissues. Although a large number of HPLC and CE papers have been published on nucleic acid constituent applications, relatively little has been written on the mechanisms of the separations. However, to optimize analytical conditions knowledgeably and rapidly, it is important to know why and how these separations occur and the factors that affect them. The HPLC methods for the analysis of nucleic acid constituents and the information available on some of the mechanisms of separation of nucleotides, nucleosides and their bases, as well as the analysis of these compounds by CE and the factors that affect these separations are discussed.     

CE frontal analysis based on simultaneous UV and contactless conductivity detection: a general setup for studying noncovalent interactions

CE frontal analysis (CE-FA) has been established as a powerful tool to study noncovalent interactions between macromolecules and small molecules such as drug substances or pharmaceutical excipients. However, when using traditional commercial CE instrumentation, a serious drawback is related to the fact that only UV-active compounds can be studied. In recent years, contactless conductivity detection has become an attractive alternative to UV detection in CE due to its high versatility. In this study, we combine contactless conductivity detection and UV detection in a highly versatile setup for profiling noncovalent interactions between low-molecular-weight molecules and macromolecules. In the case of molecules having a chromophore the setup allows determination of binding constants using two independent detectors. The new contactless conductivity detection cell is compatible with commercial CE instrumentation and is therefore easily implemented in any analysis laboratory with CE expertise.     

毛细管电泳分析中的富集技术及其应用

毛细管电泳（CE）具有分析速度快、分离效率高、样品消耗少、成本低廉等优点,已被应用于无机离子、有机小分子、蛋白质、核酸及细胞等的分析中。CE中最常用的检测方式是紫外检测（UV）,但由于常规进样样品体积小、检测光程短,CE-UV的灵敏度往往不能满足复杂样品中痕量物质直接分析的要求。CE中的在柱富集技术包括堆积、动态pH界面、吹扫和瞬间等速电泳等,可在很大程度上提高CE-UV的检测灵敏度;另外,固相和液相微萃取技术及其与在柱富集技术相结合应用在CE中也能净化样品基质,进一步提高富集倍数,改善分析灵敏度,从而拓宽了CE-UV在复杂样品分析中的应用范围。     

Overview of the status and applications of capillary electrophoresis to the analysis of small molecules

The status of capillary electrophoresis (CE) in the analysis of small molecules is reviewed and summarised with the illustrative use of recent literature references. Examples are cited in this review which demonstrate that CE is now a recognised and established technique in many industries, law courts and government regulatory agencies. Each of the principal areas of CE application in small molecule analysis are covered in sections which highlight the recent developments and possibilities within that area. Application areas include the analysis of pharmaceuticals, agrochemicals, chiral separations, and forensics is covered. This is an update to a previous review article [J. Chromatogr. A 856 (1999) 443] and covers papers published between 1999 and 2002. Technical developments and improvements, such as the advent of capillary array instrumentation for increased sample throughput, and improved detection options are described. Overall it is concluded that CE has become a recognised and established technique in many areas and is still within a period of development of both instrumentation and application which will continue to expand usage.     

Recent advances in the application of capillary electromigration methods for food analysis

This article reviews the latest developments in the application of capillary electromigration methods for the analysis of foods and food components. Nowadays, methods based on CE techniques are becoming widely used in food analytical and research laboratories. This review covers the application of CE to analyze amino acids, biogenic amines, peptides, proteins, DNAs, carbohydrates, phenols, polyphenols, pigments, toxins, pesticides, vitamins, additives, small organic and inorganic ions, chiral compounds, and other compounds in foods, as well as to investigate food interactions and food processing. The use of microchips as well as other foreseen trends in CE analysis of foods is discussed. Papers that were published during the period June 2002-June 2005 are included following the previous review by Frazier and Papadopoulou (Electrophoresis 2003, 24, 4095-4105).     

Recent advances in the application of capillary electromigration methods for food analysis

This review covers the application of capillary electromigration methods to analyze foods and food components, including amino acids, biogenic amines, peptides, proteins, DNAs, carbohydrates, phenols, polyphenols, pigments, toxins, pesticides, vitamins, additives, small organic and inorganic ions, chiral compounds, and other compounds in foods, as well as those applications of CE for monitoring food interactions and food processing. The use of microchips as well as other foreseen trends in food analysis by CE are discussed. Papers that were published during the period June 2005-March 2007 are included following the previous review by Cifuentes (Electrophoresis 2006, 27, 283-303).     

Recent novel MEKC applications to analyze free amino acids in different biomatrices: 2009-2010

This report intends to provide an updated overview of the most important methodological developments of MEKC in the field of qualitative/quantitative analysis of free amino acids in different matrices. A good number of articles published in the years 2009-2010 addresses the main applications of such procedures together with their advantages and/or drawbacks. The usefulness of chiral CE selective methods for the separation of D-amino acids in biological and food samples and the use of microchips, as well as other foreseen trends in different areas, are also discussed.     

Capillary electrophoresis-based immunoassays: principles and quantitative applications

The use of CE as a tool to conduct immunoassays has been an area of increasing interest over the last decade. This approach combines the efficiency, small sample requirements, and relatively high speed of CE with the selectivity of antibodies as binding agents. This review examines the various assay formats and detection modes that have been reported for these assays, along with some representative applications. Most CE immunoassays in the past have employed homogeneous methods in which the sample and reagents are allowed to react in solution. These homogeneous methods have been conducted as both competitive binding immunoassays and as noncompetitive binding immunoassays. Fluorescent labels are most commonly used for detection in these assays, but enzyme labels have also been utilized for such work. Some additional work has been performed in CE immunoassays with heterogeneous methods in which either antibodies or an analog of the analyte is immobilized to a solid support. These heterogeneous methods can be used for the selective isolation of analytes prior to their separation by CE or to remove a given species from a sample/reagent mixture prior to analysis by CE. These CE immunoassays can be used with a variety of detection modes, such as fluorescence, UV/Vis absorbance, chemiluminescence, electrochemical measurements, MS, and surface plasmon resonance.