A Series of Collaborations between Various Pharmaceutical Companies and Regulatory Authorities Concerning the Analysis of Biomolecules Using Capillary Electrophoresis: Additional Instruments/Buffer

An international project team (including members from US, Canada and UK) was formed from a number of interested biopharmaceutical companies and regulatory authorities to conduct a cross-organisation collaboration exercise. The results of the first comparison with eight different organisations that used instruments of the same equipment model, the same reagents, and the same methodology has been reported previously [1]. This report represents the addition of other instruments using a different run buffer. The relative migration times were different, as expected, prohibiting a direct comparison between companies. The within-organisation variability was low for both relative migration time (<0.34% RSD% for all companies save one) and the peak area (<5% RSD% for all companies save one) when measuring the purity of a representative IgG sample. The apparent molecular weight of bovine serum albumin was measured with good precision (less than 10% RSD% across all companies) to the theoretical value when all data is utilized (67.5 kDa compared to 66.4 kDa). For a representative IgG sample, the three main components, IgG Light Chain, IgG Non-glycosylated Heavy Chain, and IgG Heavy Chain, could not be separated, specifically the IgG Non-glycosylated Heavy Chain and IgG Heavy Chain. When the IgG Non-glycosylated Heavy Chain and IgG Heavy Chain were combined for all organisations, the fractional peak area for the IgG Light Chain and IgG Non-glycosylated Heavy Chain + IgG Heavy Chain peak also showed excellent agreement, with less than 7.5 and 3.5% RSD%, respectively. The value of this exercise is in demonstrating the reliability of CE for the determination of apparent size of biopharmaceutical proteins. This underpins the appropriate use of such CE data in support of regulatory submissions.

     

A Series of Collaborations Between Various Pharmaceutical Companies and Regulatory Authorities Concerning the Analysis of Biomolecules Using Capillary Electrophoresis

An international project team (including members from US, Canada and UK) has been formed from a number of interested biopharmaceutical companies and regulatory authorities to conduct a cross-organisation collaboration exercise. The results from this exercise demonstrate the robustness of CE-SDS across eight different organisations that used instruments of the same equipment model, the same reagents, and the same methodology. Data generated from the analysis of a series of molecular weight markers showed very good precision with regards to relative migration time (RMT) both within and between organisations. The apparent molecular weight of bovine serum albumin (BSA) was measured with good precision to within approximately 2% RSD across the participants. A representative IgG sample showed similar results with regards to relative migration time of its 3 main components, IgG Light Chain, IgG Non-glycosylated Heavy Chain, and IgG Heavy Chain. Fractional peak area for each peak also showed good agreement, with less than 9% RSD for all peaks. This exercise will facilitate both increased regulatory and industrial opinion of CE for biopharmaceutical analysis.CE in the Biotechnology & Pharmaceutical Industries: 7th Symposium on the Practical Applications for the Analysis of Proteins, Nucleotides and Small Molecules, Montreal, Canada, August 12–16, 2005     

毛细管电泳技术在药物分离分析中的研究与应用

综述了近5年毛细管电泳在手性药物拆分、药物制剂及中草药分析中的应用.在手性药物拆分的应用中主要探讨了手性选择剂的种类及毛细管分离方法;在药物制剂、中草药的应用中主要介绍该法对药效成分进行的分离及定量分析,总结方法的检出限、线性范围和检测方法;最后,探讨了毛细管电泳在求取药物水解常数上的应用.提出毛细管电泳在药物分析中将有广阔的应用前景.     

碱性蛋白质毛细管电泳分离研究

碱性蛋白质毛细管电泳分离研究任吉存，邓延倬，程介克（武汉大学化学系分析测试科学系，武汉，４３００７２）关键词毛细管电泳，碱性蛋白质，吸附，精氨酸，赖氨酸蛋白质的吸附作用严重影响了毛细管电泳分离蛋白质的重现性［１］．为此，人们寻找各种途径来克服蛋白质的...     

以L-谷氨酰胺为手性选择剂毛细管电泳拆分几种药物旋光对映体

以Ｌ－谷氨酰胺为手性选择剂,建立了对普萘洛尔、慢心律、心律平、苯丙哌酮、异丙肾上腺素、氯胺酮、山莨菪碱、洛贝林、布比卡因、苯海索等１０种手性药物对映体的毛细管电泳快速分离方法,在含有６０ｍｍｏｌ／Ｌ Ｌ－谷氨酰胺,６０ｍｍｏｌ／Ｌ硼砂（ｐＨ＝９．２５）的缓冲溶液中,１０种手性药物在１１ｍｉｎ内达到基线分离。并对分离机理作了初步探讨。     

电中性分子间相互作用分析的毛细管电泳方法研究

使用具有负有效淌度的十二烷基磺酸钠胶束赋予中性分子有效淌度,以中性主体分子18-冠-6和中性客体分子对-硝基苯胺形成中性超分子包结化合物为实例,建立了中性分子相互作用的毛细管电泳分析方法,发现对-硝基苯胺和18-冠-6与SDS的相互作用存在相当的差异.测得18-冠-6和对-硝基苯胺的lgK为4.14,18-冠-6与2-氨基吡啶、18-冠-6与SDS以及SDS与2-氨基吡啶的lgK分别为3.65,3.13和1.15.     

用N,N-二甲基六烷基溴化铵涂层的毛细管对重组促红细胞生长素及其酶解产物进行毛细管电泳和毛细管电泳-电喷雾质谱分析

采用毛细管电泳技术研究了重组促红细胞生长素(rhEPO)的分离问题。用N,N-二甲基六烷基溴化铵（6,6-ionene）涂层的毛细管测定了rhEPO中唾液酸的微多相性,同时采用毛细管电泳-质谱(CE-MS)联用技术在22 min内鉴定了rhEPO 20段胰酶消化肽中的11段。该方法简单快捷,重现性好,可用于蛋白质一级结构的测定。     

人工神经元网络法提高毛细管电泳药物控制分析准确度的研究

研究了人工神经元网络法在毛细管电泳定量测定memantine中提高测定准确度 的可行性。在毛细管电泳法定量测定memantine的过程中，其浓度与峰高或峰面积 以及与二者和内标的比值均没有良好的线性关系。人工神经元网络具有很强的非线 性校正能力，其最大优点是无须对分离体系及组分的迁移行为预先予以了解。人工 神经元网络的输为memantine的峰高和峰面积，输出为memantine的浓度。通过实验 确定的网络结构为2：1：1型。由于人工神经元网络的通用性，该法也可用于毛细 管电泳在其他药物控制分析中改善定量分析的准确度。     

葡萄糖氧化酶光敏氧化反应过程的毛细管电泳法监测

生物学研究表明，叶琳和酞著衍生物对癌变组织具有亲和性[1]．此类物质在吸收特定波长的光之后产生活性氧，可光敏氧化生物分子，并杀死癌细胞．光动力疗法对脑癌、颈癌和服癌等浅层癌具有明显的疗效[2]，研究各种光敏剂与生物大分子的作用机理[3，4]，对筛选新型光疗药物，改进光疗效果具有重要价值．癌细胞中葡萄糖的代谢非常旺盛，若阻断这种代谢，有可能实现对癌细胞进行代谢调控的目的．因此，本文选用葡萄糖氧化酶为模型化合物，用毛细管电泳法监测了4种新型光敏剂[ZnPcS4，ZnPc（NHCH2COOH）4，C1A1PcS，Por－Phyrine－like]…     

Investigation of the Interaction Between Lidocaine and the Components of Hyaluronic Acid Using Frontal Analysis Continuous Capillary Electrophoresis

The frontal analysis continuous capillary electrophoresis (FACCE) technique was used for the characterziation of the interaction between lidocaine-HCl (Lido) and components of Na-hyaluronic acid (HA). N-Acetylglucosamine (GlcNAc) and glucuronic acid (GluA) were the components of Na-hyaluronic acid. For the investigations fused silica capillaries were used. The FACCE method was compared to affinity capillary electrophoresis (ACE). The association constants between lidocaine-HCl and the components of Na-hyaluronic acid were determined using FACCE. It was observed that only an interaction between Lido and GluA exhibited. The association constant (K _lido-GluA) between Lido and GluA was 26 ± 0.3 L mol^?1.     

Investigation of the Interaction Between Lidocaine and the Components of Hyaluronic Acid Using Frontal Analysis Continuous Capillary Electrophoresis

The frontal analysis continuous capillary electrophoresis (FACCE) technique was used for the characterziation of the interaction between lidocaine-HCl (Lido) and components of Na-hyaluronic acid (HA). N-Acetylglucosamine (GlcNAc) and glucuronic acid (GluA) were the components of Na-hyaluronic acid. For the investigations fused silica capillaries were used. The FACCE method was compared to affinity capillary electrophoresis (ACE). The association constants between lidocaine-HCl and the components of Na-hyaluronic acid were determined using FACCE. It was observed that only an interaction between Lido and GluA exhibited. The association constant (K _lido-GluA) between Lido and GluA was 26 ± 0.3 L mol⁻¹.

     

毛细管电泳法测定猪体组织中甜菜碱含量

建立了毛细管电泳法快速测定猪体组织中甜菜碱含量的方法。甜菜碱首先转化为苯甲酰甲基酯后直接上样测定。pH为 3 .0的磷酸缓冲溶液使甜菜碱酯化物和甜菜碱结构类似物酯化物之间以及酯化物和酯化剂之间都能很好地分离 ,这也省去了反应混合液的前处理。该法标准曲线的线性范围为 4～ 6 0 0mg L ,相关系数r为 0 .9999,最低检测限为 1mg L ,相对标准偏差为 2 .2 %～ 4.7% ,标准加入回收率为 95 9%～ 98.4%     

A Novel Affinity Capillary Electrophoresis for the Detection of Gene Mutation Using Immobilized Oligonucleotides-Polyacrylamide Conjugate

It has been found that small mutations of certain genes are the definitive origin of many heritable disorders and cancers with striking development of recent molecular biology. Such new findings have taken close-up of the importance of gene mutation assays based on the difference of DNA base sequences in diagnostic or medical field Capillary electrophoresis can be a good candidate for an ideal method on such gene analysis, because the methods can be performed with trace amount of samples, high resolution and shorter running time. We have established an effect of oligonucleotide, which was introduced onto capillary inner surface, on the recognition of an overall sequence of sample DNA fragments as an affinity ligand.     

Recent Applications of Capillary Electrophoresis in the Determination of Active Compounds in Medicinal Plants and Pharmaceutical Formulations

The present review summarizes scientific reports from between 2010 and 2019 on the use of capillary electrophoresis to quantify active constituents (i.e., phenolic compounds, coumarins, protoberberines, curcuminoids, iridoid glycosides, alkaloids, triterpene acids) in medicinal plants and herbal formulations. The present literature review is founded on PRISMA guidelines and selection criteria were formulated on the basis of PICOS (Population, Intervention, Comparison, Outcome, Study type). The scrutiny reveals capillary electrophoresis with ultraviolet detection as the most frequently used capillary electromigration technique for the selective separation and quantification of bioactive compounds. For the purpose of improvement of resolution and sensitivity, other detection methods are used (including mass spectrometry), modifiers to the background electrolyte are introduced and different extraction as well as pre-concentration techniques are employed. In conclusion, capillary electrophoresis is a powerful tool and for given applications it is comparable to high performance liquid chromatography. Short time of execution, high efficiency, versatility in separation modes and low consumption of solvents and sample make capillary electrophoresis an attractive and eco-friendly alternative to more expensive methods for the quality control of drugs or raw plant material without any relevant decrease in sensitivity.     

Multiresidue Analysis of Pesticides in Environmental Waters by Capillary Electrophoresis Using Simultaneous UV and Electrochemical Detection

A rapid multiresidue method has been developed for the analysis of eight pesticides (triazines, triazinones and ureas) in environmental waters. A simple end‐column electrochemical detector was used in combination with an available commercial capillary electrophoresis instrument with UV detection. The determination of these pesticides using micellar electrokinetic capillary chromatography (MEKC) with dual electrochemical and UV detection is the first time reported. In both detection systems, a linear range was obtained for the eight pesticides, concentrations lower than 5.0×10^?5 mol L^?1, in 0.020 mol L^?1 boric acid at pH 8.3 and containing 0.025 mol L^?1 of sodium dodecylsulfate, to obtain selectivity in the additional separation by micellar distribution process. Under these conditions a lower detection limit than 2.0×10^?6 mol L^?1 (0.15 pmol of pesticide) was achieved for the most of them. The eight pesticides are resolved in less than 14 min.     

毛细管电泳-激光诱导荧光法分离检测DNA片段及基因扩增产物

以羟丙甲基纤维素和非交联聚丙烯酰胺浴液为筛分介质,将毛细管电泳-激光诱导荧光法用于DNA片段及基因扩增产物的分离检测。探讨了非胶筛分介质中高分子化合物的浓度、电解质的浓度、内插试剂用量等对DNA片段分离检测的影响;考察了DNA片段迁移时间和峰面积的重现性及DNA片段定量检测的关系。建立了一种快速、灵敏的DNA片段及基因扩增产物分离检测方法。     

毛细管电泳-柱端安培检测中分离毛细管与检测电极对接方式比较的研究

针对目前毛细管电泳中普遍采用的水平式和直立式两种柱端安培检测形式,在各种可能的对接角度下,以邻苯二酚、间苯二酚和对苯二酚3种二酚作为分离分析测试对象,对分离毛细管与检测电极的对接方式进行了优化比较,并对每一种对接方式的分析特性进行了测试。结果表明：在以3种典型的方式对接时,分离毛细管与检测电极为倒直立式对接时,获得的检测灵敏度最高,响应电流最大,水平式次之,直立式最小。     

Rapid Capillary Electrophoresis Method for Simultaneous Determination of Abemaciclib,Ribociclib, and Palbociclib in Pharmaceutical Dosage Forms: A Green Approach

Advances in the treatment of HR+/HER2- breast cancer phenotype have been made with the introduction of abemaciclib, ribociclib, and palbociclib, inhibitors of cyclin D dependent kinases 4 and 6 (CDK4/6). Here, a novel, fast, cheap, and green CE method for the simultaneous determination of these three CDK4/6 inhibitors in less than 4 min is proposed for the first time. Separation was achieved by capillary zone electrophoresis in an acidic medium, in accordance with the structures of the analytes and their pKa values. The optimal pH of the running buffer was found to be 2.9. The optimal method conditions were 27.5 kV separation voltage, 30 °C, 5 s injection time under 50 mbar pressure, and 50 mM phosphate background buffer with benzimidazole as an internal standard. The developed method was validated with respect to robustness, selectivity, accuracy, precision, linearity, and limits of detection. The method was shown to be linear in the range of 10 to 100 µg mL⁻¹ with correlation coefficients higher than 0.9981. A greenness assessment of the proposed method was performed, and the method was shown to be green. The validated method was successfully applied to pharmaceutical dosage forms of all CDK4/6 inhibitors.     

Capillary Zone Electrophoresis Investigation of Interactions between Granulocyte-colony Stimulating Factor and Dextran Sulfate／Carrageenan Oligosaccharide

Granulocyte-colony stimulating factor (G-CSF) is an O-glycosylated glycoprotein. G-CSF stimulates the proliferation and differentiation of hematopoietic progenitor cells committed to the neutrophil/granulocyte lineage in a dose-dependent manner. The most important application of G-CSF is in the treatment of transient phases of leukopenia following chemotherapy and/or radiotherapy. Using capillary zone electrophoresis (CZE), former studies showed that G-CSF could interact with polysulfa…