毛细管电泳的原理及应用(第五讲)毛细管电泳在医药领域中的应用

简单介绍了毛细管电泳在医药领域中的应用，包括药物分析、中药成分分析、手性对映体分离分析和临床化学、法医及单细胞分析等。药物分析中包括主药成分分析、相关杂质检测、药物计量离子配比测定和定量分析等。中药成分分析中包括各类药效成分、中药材中主要成分及中药复方制剂成分分析。手性对映体分离中包括机理研究、新型手性选择剂。临床化学中包括临床疾病诊断、临床蛋白分析、临床药物监测、药物代谢研究和分子生物学测定。法庭科学中包括毒物分析、枪击残余物分析、炸药分析、笔迹墨水分析。单细胞分析中包括神经细胞分析、红细胞分析和胚胎细胞分析。     

毛细管电泳法在手性分离中的应用及进展

近年来,毛细管电泳(CE) 手性分离方法的研究主要集中在各种手性添加剂与对映体药物的匹配及实验条件的最优化选择上.目前,较为成熟的CE分离模式有:区带电泳(CZE)、凝胶电泳(CGE)、等速电泳(CITP)、胶束电动色谱(MEKC)和非水电泳(NACE)等,并已成功地用于手性化合物对映体的分离.CE手性分离研究正朝着新型手性选择剂的研制和实现与其他各种定性分析仪器及其他色谱分离模式的联用方向发展.     

气相色谱手性固定相进展

1966年Gil-Av等人首次用手性固定相在毛细管柱上分离了氨基酸对映体。至今这种方法已发展为研究和分离对映体的一种重要技术,并愈来愈多地用于天然产物绝对构象(外激素、香料成分等)的测定,不对称选择合成中对映体纯度及过剩量测定,手性药物中对映体纯度的测定,手性化合物在化学转移机理等方面的研究。目前气相色谱中用于分离对映体的     

手性毛细管电泳实验中手性选择剂取代度的影响研究

针对手性毛细管电泳实验“手性药物氧氟沙星对映体的分离检测”的教学过程中,发现手性选择剂取代度对实验结果具有重要影响,详细研究了一系列不同取代度的羟丙基-β-环糊精对氧氟沙星对映体分离效果的影响。研究结果表明：对映体的分离度随取代度变化显著,且取代度DS=7.9时分离度最佳。依据取代度的测定方法,对市售羟丙基-β-环糊精的质量进行评估,以确保手性毛细管电泳实验结果的稳定性和良好的教学效果。     

5种手性药物毛细管电泳对映体拆分

采用α、β、γ－环糊精和二甲基、羟丙基－β－环糊精等６种中性环糊精为手性添加剂,在毛细管电泳分离中,进行了５种手性中心具有相似特点的碱性药物对映体的拆分研究。详细地讨论了环糊精类型及其浓度、pＨ值、分离电压、操作温度、以及有机添加剂等操作参数对药物对映体拆分的影响。     

毛果芸香碱对映体的毛细管电泳手性分离

采用β-环糊精及其衍生物作为手性选择剂对毛果芸香碱对映体进行了分离,研究了环糊精类型和浓度对分离的影响,同时考察了背景电解质pH、操作电压和温度等因素对对映体分离的影响;结果表明,采用羟丙基-β-环糊精(HP-β-CD)可以使毛果芸香碱对映体达到基线分离,优化条件下手性分离度可达2．79,为此类药物提供了一种简便、快速的毛细管电泳手性分离分析方法。     

使用二元环糊精体系毛细管电泳法分离手性药物

采用高效毛细管区带电泳法，分别以羟丙基-β-环糊精（HP-β-CD）和高磺化-β-环糊精（HS-β-CD）及二者混合物为手性选择剂，研究了5种药物的对映体分离，并取得很好的对映体分离结果。比较了HP-β-CD和HS-β-CD手性识别能力，分析了二元环糊精体系的“协同效应”。     

MAH-β-CD的合成及其在毛细管电泳手性拆分中的应用

合成了顺丁烯二酸酐-β-环糊精(MAH-β-CD),并通过红外光谱、质谱对其结构进行表征。以20 mmol/L乙酸铵为缓冲溶液,MAH-β-CD作为手性选择剂,利用毛细管电泳对氨基酸和手性药物对映体进行拆分研究。考察了缓冲溶液pH、分离电压和手性选择剂浓度等对拆分效果的影响,在优化条件下,成功拆分了3种氨基酸(DL-甲硫氨酸、DL-精氨酸、DL-赖氨酸)和两种手性药物(沙丁胺醇、氯美扎酮)对映体,分离度分别为5.11,5.55,2.99,2.33和1.64。     

8种芳香醇对映体的毛细管区带电泳手性拆分及其光学纯度分析

采用毛细管电泳(CE)手性分离方法对8种不同取代基的芳香醇对映体样品的光学纯度进行分析。通过对手性选择剂β-CD-NH3Cl的浓度、背景电解质的浓度、pH值及操作电压等影响因素的研究,优化了分离条件,成功地对8种芳香醇对映体样品进行了手性拆分;同时,对8种对映体样品进行了光学纯度检查,并与HPLC测定结果进行比较。结果表明,3批样品中对映体过量(e.e.%)测定值与HPLC法的测定结果相一致。该方法简单、准确,可用于该8种不同取代基芳香醇的手性拆分和e.e.%值的测定。     

毛细管电泳在手性化合物分离中的应用进展

由于手性化合物尤其是手性药物的两个对映体具有不同的化学性质和生理活性,对手性化合物进行分离在医药、生物、食品和环境等领域都具有十分重要的意义。毛细管电泳由于其独特的优势已广泛应用于手性物质的分离。本文对2013~2015年毛细管电泳用于手性分离的最新进展进行了综述,并对其发展前景进行了展望。     

毛细管电泳用于药物分析的研究进展

药物分析是毛细管电泳（CE）的重要应用领域,所有CE分离模式与检测方法都在各种药物及其不同形式样品的分离分析中显示出特色和应用能力。该文从药品分析领域中的小分子药物（包括手性药物）及其有关物质、中药与天然产物、体内药物分析、生物制品药物分析等几个方面,综述了近几年CE在这些传统药物分析领域应用的研究进展。限于篇幅,未包括现代药物分析研究比较活跃的理化常数测定、亲和毛细管电泳与结合常数研究（药物与受体间的相互作用等）、临床生物标志物分析、代谢组学和微流控芯片CE分析等方面的内容。根据目前传统药物分析领域的发展,该文关注到近期CE在顺应药物分析的法规需求、电容耦合非接触电导检测（CE-C⁴ D）、改进检测灵敏度与精密度、CE-十二烷基硫酸钠（SDS）毛细管电泳、全柱成像毛细管等电聚焦（icIEF）、抗体分析等方面的新进展。该文结合文献,讨论了目前传统药物分析领域的需求,以及CE在其中的地位、挑战和机遇。对目前CE主要作为互补分析方法在化学药和中药分析中的应用研究提出了一些针对性的建议,期待CE在生物制品分析中的特色和能力得到进一步的发挥,同时提出CE-MS和对CE分析重复性改进等新进展可能对未来CE应用领域的大幅度扩展。该综述主要涉及近3年（2017年1月到2020年2月）及部分2016年的相关文献。     

A decade of capillary electrophoresis

Since the introduction of the first commercial capillary electrophoresis (CE) instrument a decade ago, CE applications have become widespread. Today, CE is a versatile analytical technique which is successfully used for the separation of small ions, neutral molecules, and large biomolecules and for the study of physicochemical parameters. It is being utilized in widely different fields, such as analytical chemistry, forensic chemistry, clinical chemistry, organic chemistry, natural products, pharmaceutical industry, chiral separations, molecular biology, and others. It is not only used as a separation technique but to answer physicochemical questions. In this review, we will discuss different modes of CE such as capillary zone electrophoresis, micellar electrokinetic chromatography, capillary gel electrophoresis, capillary isoelectric focusing, and capillary electrochromatography, and will comment on the future direction of CE, including array capillary electrophoresis and array microchip separations.     

Generic capillary electrophoresis conditions for chiral assay in early pharmaceutical development

Generic capillary electrophoresis (CE) conditions have been implemented for chiral separations in early pharmaceutical development. The chiral CE separations of several pharmaceutical samples at different stages of development, i.e., discovery, process chemistry, and investigative new drug application, have been obtained using sulfated beta-cyclodextrin (CD). Several sulfated beta-CDs have been screened to select an appropriate enantioselective agent. The use of a generic CE method allows for a convenient and rapid chiral recognition of different weak bases, with minimal or no method development. CE using sulfated beta-CD for the chiral separation of N-benzoyl methyl piperazine has been validated for linearity, precision, accuracy, limits of detection and quantitation (LOD, LOQ). Although less sensitive than a specific liquid chromatography method using a Chiralpak AD column, the overall performance of the chiral CE method was found comparable. Validation data demonstrate that a LOD of 0.1%, sufficient to fulfill regulatory requirements, is achievable by chiral CE.     

Recent advances of capillary electrophoresis in pharmaceutical analysis

This review covers recent advances of capillary electrophoresis (CE) in pharmaceutical analysis. The principle, instrumentation, and conventional modes of CE are briefly discussed. Advances in the different CE techniques (non-aqueous CE, microemulsion electrokinetic chromatography, capillary isotachophoresis, capillary electrochromatography, and immunoaffinity CE), detection techniques (mass spectrometry, light-emitting diode, fluorescence, chemiluminescence, and contactless conductivity), on-line sample pretreatment (flow injection) and chiral separation are described. Applications of CE to assay of active pharmaceutical ingredients (APIs), drug impurity testing, chiral drug separation, and determination of APIs in biological fluids published from 2008 to 2009 are tabulated.     

Determination of enantiomeric excess by capillary electrophoresis

Capillary electrophoresis (CE) is becoming an established method for the determination of chiral trace impurities. This paper provides an overview of the state of the art of CE for such determinations. Detection limits of 0.1% impurity is widely accepted as a minimum requirement for chiral trace impurity determinations. This can be relatively easily achieved with CE. However, determination of lower concentrations requires careful optimization of the separation system. Four factors that are of particular significance for trace enantiomeric determinations: resolution, limit of detection, linear range and type of detection, are discussed. Further, the advantages and disadvantages of derivatization in this context are treated as well as the separation approach, ie., direct chiral separation or separation after the formation of diastereomers. It is concluded that the limit of impurity detection can be about 0.05% when UV detection is employed. Using laser-induced fluorescence detection, a quantitative determination at the 0.005% level is often possible.     

Enantiomeric impurity determination in capillary electrophoresis using a highly-sulfated cyclodextrins-based method

Capillary electrophoresis (CE), using highly-sulfated cyclodextrins as chiral selectors, has been applied to determine the chiral purity of pharmaceutical compounds. A chiral separation strategy, developed earlier for racaemic mixtures, was applied on four basic drugs (propranolol, atenolol, chlorpheniramine and tryptophan methylester). The aim was to develop validated separation methods which allow determination of 0.1% impurity levels of the unwanted enantiomers (distomer) in the presence of 99.9% of the active compound (eutomer). The linearity, quantification limits for the trace enantiomers and the precision of the measurements were determined. In a second part, impurity separations have been simulated in order to evaluate the required resolution when assaying impurities. It is shown that a baseline resolution of 1.5, generally accepted for racaemic mixtures, does not always allow good impurity determinations. Two alternative methods to solve this problem have been proposed.     

毛细管电泳的原理和应用(第六讲)CE在DNA、糖和离子分析中的应用

 简要介绍了ＣＥ在ＤＮＡ、糖和离子分析中的应用。ＤＮＡ分析包括分离原理、方法及检测手段，碱基、核苷和核苷酸分析，纯度检测和微量制备，ｓｓＤＮＡ和ｄｓＤＮＡ分析，基因突变检测及ＤＮＡ序列分析等。糖分析包括衍生化技术、检测方法、所用模式及糖复合物的分析等。离子分析包括阴离子分析原理、检测方法、影响因素、应用及阳离子分析等。     

Chemiluminescence detection coupled to capillary electrophoresis

In recent years, chemiluminescence (CL)-based detection coupled to capillary electrophoresis (CE) as separation technique has attracted much interest due to new advances in home-made configurations, sample-treatment techniques for application to real matrixes, development of a commercial instrument and use of miniaturization techniques to obtain micro total analysis systems incorporating CE separation and CL detection in microchips. We present some developments, key strategies and selected analytical applications of CE-CL since the year 2000 in diverse fields (e.g., clinical and pharmaceutical, environmental or food analysis).     

2019年毛细管电泳技术年度回顾

本文归纳了ISI Web of Science中检索的2019年度毛细管电泳（CE）技术的相关论文,从生物分析、药物分析、临床检验及医学诊断、手性拆分、食品检测、其他化合物和离子检测以及毛细管电泳-质谱（CE-MS）的应用等7个方面进行了分类说明;简要介绍了2019年度与CE技术有关的国际会议和国内会议及各会议的重点研究报告。