Capillary electrophoresis with electrochemiluminescence detection for the analysis of quinolone drugs and pharmacokinetics study

A novel method for the determination of two quinolone drugs norfloxacin （NOR） and levofloxacin （LVX） was described by capillary electrophoresis with electrochemiluminescence detection. The good relationship （r ≥ 0.9991） between peak area and concentration of analytes was established over two orders of magnitude. The limits of detection （LOD, S/N = 3） in standard solution are 4.8 × 10^-7 mol/L for NOR and 6.4 × 10^-7 mol/L for LVX, respectively. The limits of quantitation （LOQ, S/N = 10） in real human urine samples are 1.2 × 10^-6 mol/L for NOR and 1.4 × 10^-6 mol/L for LVX, respectively. The present method was successfully applied to the determination of NOR and LVX in human urine and the studv of oharmacokinetics of NOR.

Analysis of Amino Acids in a Single Human Red Blood Cell by Capillary Zone Electrophoresis with Intracellular NDA- derivatization and Electrochemical Detection

A novel method for determination of amino acids in individual human red blood cells has been developed. In this method, the derivatization reagents (NDA and CN-) are introduced into living cells by electroporation. After completion of derivatization, the amino acids in a single cell is determined by capillary zone electrophoresis with end-column amperometric detection.

Quality Analysis of Herbal Medicine Products Prepared from Herba Sarcandrae by Capillary Electrophoresis with Electrochemical Detection

A capillary electrophoresis with electrochemical detection（CE-ED） method was developed for the quality analysis of herbal medicine products prepared from the same herb of Herba Sarcandrae： Fufang Caoshanhu tablets, Qingrexiaoyanning capsules, and Xuekang oral liquids. Under the optimal analysis conditions, the low detection limit[l.0×10^-7 mol/L（S/N=3）] and the wide linear range（1.0×10^-7-1.0×10^-4 mol/L） were obtained for quality standard compound of isofraxidin. The precisions of the peak current and the migration time（as RSDs） for the real sample analysis were 2.0%-2.6%, and 1.2%-1.8% for isofraxidin, respectively. The contents of isofraxidin detected were 15.77 μg/tablet, 0.48 mg/capsule, 1.2 mg/ampoule（Jiangxi）, and 0.44 mg/ampoule（Dalian） for Fufang Caoshanhu tablets, Qingrexiaoyanning capsules, and Xuekang oral liquids from different manufacturers, respectively. Quality estimate was conducted by comparing the contents of isofraxidin in the herbal medicine products with the demanded values of Chinese pharmacopeia. In addition, based on their own unique CE-ED profiles（namely, CE-ED electropherograms） the Xuekang oral liquids from the different manufacturers could be easily identified.

Study on the Interaction between Strychnine and Bovine Serum Albumin by Capillary Electrophoretic Frontal Analysis

The protein binding constant, binding sites of the Strychnos alkaloid-strychnine and bovine serum albumin （BSA） was determined by capillary electrophoretic frontal analysis （CE-FA） for the first time. The experiment was carried out in a polyacrylamide-coated fused silica capillary （48.4 cm×50 μm i.d., 38.1 cm effective length） with 20 mmol/L citrate/MES buffer （pH 6.0, ionic strength 0.17）. The applied voltage was 12 kV and detection wavelength was set at 257 nm. The plateau height of the peak was employed to determine the unbound concentration of drug in BSA equilibrated sample solution based on the external drug standard in the absence of protein. The present method provides a convenient, accurate technique for the early stage of drug screening.

Highly sensitive trivalent copper chelate-luminol chemiluminescence system for capillary electrophoresis chiral separation and determination of ofloxacin enantiomers in urine samples

A simple,fast and sensitive capillary electrophoresis（CE） strategy combined with chemiluminescence（CL） detection for analysis of ofloxacin（OF） enantiomers was established in the present work.Sulfonated p-cyclodextrin（β-CD） was used as the chiral additive being added into the running buffer of luminol-diperiodatocuprate（Ⅲ）（K[Cu（HIO6）2],DPC） chemiluminescence system.Under the optimum conditions,the proposed method was successfully applied to separation and analysis of OF enantiomers with the detection limits（S/N=3） of 8.0 nM and 7.0 nM for levofloxacin and dextrofloxacin,respectively.The linear ranges were both 0.010-100 μM.The method was utilized for analyzing OF in urine;the results obtained were satisfactory and recoveries were 89.5-110.8%,which demonstrated the reliability of this method.This approach can also be further extended to analyze different commercial OF medicines.

Determination of Amino Acids in an Individual Erythrocyteby Capillary Electrophoresis with Intracellular FITC-derivatization and Laser-induced Fluorescence Detection

A novel approach for analysis of amino acids in individual erythrocytes was established. In this method, the derivatization reagent was introduced into the living cells by electroporation. After derivatization, the amino acids in a single cell were determined by capillary electrophoresis with laser-induced fluorescence detection.

Rapid Separation and Quantification of Iron in Uranium Nuclear Matrix by Capillary Zone Electrophoresis (CZE)

A method was developed for rapid separation and determination of iron by employing capillary zone elec-trophoresis (CZE) technique with direct UV detection. Iron could be separated from matrix uranium by di-rect injection of dissolved sample solution into capillary using a mixture of 10 mM HCl and 65 mM KCl (pH = 2) as background electrolyte (BGE) at an applied voltage of 15 kV. The developed method has a very high tolerance for the matrix element U (100 mg/mL) and as such may not need prior separation of iron from the matrix. Iron could be separated with better than 95% recovery. The method showed a linear calibration over a concentration range 1-50 ppm of Fe. The migration times for the iron peak were reproducible within 1% for both pure Fe(III) and in presence of matrix uranium (80 mg/mL). The precision (RSD, n = 22) of peak area obtained for 1ppm of iron was 3.5%. The limit of detection (LOD) (3 ) was 0.1 ppm and the ab-solute LOD was 9 × 10-14 g considering the sample injection volume of 1.5 nL. The developed method has been validated by separating and determining iron in two certified reference materials of U3O8. The method was applied for the determination of iron in different uranium based nuclear materials. The CZE method is versatile for routine analysis as it is simple, rapid and has simple sample preparation procedure.

Quick Single Run Capillary Zone Electrophoresis Determination of Active Ingredients and Preservatives in Pharmaceutical Products

The paper deals with the development of a rapid and efficient Capillary Zone Electrophoresis (CZE) method for Quality Control analysis of pharmaceutical preparations containing antihistamines, decongestants, anticholinergic remedies and preservatives. Active ingredients of interest are: ChlorPheniramine Maleate (CPM), DiPhenhydramine Hydrochloride (DPH), Ephedrine hydrochloride (E), Isopropamide Iodide (II), Pheniramine Maleate (PM), Lidocaine hydrochloride (L), Tetracaine hydrochloride (T), Clopamide Hydrochloride (CH), DiHydroErgochristine (DHE), PhenylEphrine hydrochloride (PE) and Acetaminophen (A). Preservatives studied are: MethylParaben (MeP), EthylParaben (EtP), PropylParaben (PrP), ButylParaben (BuP), p-HydroxyBenzoic Acid (p-HBA). All these analytes were separated in a single run using 60 mM tetraborate buffer solution (TBS) pH = 9.2 as a BackGround Electrolyte (BGE) by using an uncoated fused silica capillary of I.D. = 50 mm and applying a voltage of 25 kV in the first part of the electropheretic run (up to 5.8 min) and 30 kV for the remaining time. The hydrodynamic pressurization of the inlet vial was 20 psi at 7.2 min. up to the end of analysis. Total separation time was of 7.5 min. The method was then successfully validated and applied to the simultaneous determination of active ingredients and preservatives. Good repeatability, linearity, and sensitivity were demonstrated. Precision of migration time (tm) was RSD% < 0.53% and of corrected peak area (Ac) was RSD% < 6.15%. The linearity evaluation gave 0.9928 2 1.000. LOD and LOQ, accuracy (recovery) and ruggedness were evaluated for each analyte demonstrating the good reliability of the method. Analyses of some pharmaceutical real samples were performed.

Rapid and Sensitive CZE Method for Quality Control Analysis of Pharmaceuticals Containing Pseudoephedrine, Triprolidine and Paracetamol

A rapid and sensitive CZE (Capillary Zone Electrophoresis) method for pharmaceutical analysis was developed and fully validated. The active compounds: Pseudoephedrine hydrochloride (PSE), Triprolidine hydrochloride (TRI) and Paracetamol (PAR) were separated and quantitatively determined using the tris-borate 30 mM buffer at pH = 9.0 as a Background Electrolyte (BGE). The electrophoretic separation was carried out at 25 kV in an unmodified fused silica capillary of I.D. = 50 μm with a “bubble-cell” for UV detection at 210 nm and 25&deg;C. The separation was reached in about 3 min. After calibration the method was applied for analysis of three commercially available pharmaceutical preparations. The repeatability (RSD%) of migration time (tm) was ranging between 0.47% and 0.90% and of peak areas (A) between 0.63% and 3.64%. The Limit of Detection (LOD) values was of 0.19 μg/mL, 0.31 μg/mL and 0.08 μg/mL for respectively PSE, TRI and PAR. The results obtained in this study showed that the proposed method was useful in routinely analysis of pharmaceuticals.

寡核苷酸药物生物分析方法的研究进展

寡核苷酸药物是具有不同功能化学修饰的DNA或RNA,主要在基因水平上发挥治疗的作用。到目前为止,已经批准16种寡核苷酸药物用于临床治疗,这也导致了人们对寡核苷酸药物生物分析的兴趣日益增加。尤其需要强大的生物分析工具来研究这些药物的质量控制及体内的药代动力学研究。该篇综述总结了液相色谱、毛细管凝胶电泳、杂交酶联免疫吸附技术在检测寡核苷酸药物的优缺点,为该类药物的研究及质量控制提供一定的思路。

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。

线上线下融合的仿真实验软件(电子书)研发及其教学应用——以毛细管电泳实验为例

以Web设计语言和Photoshop、Flash、3D Max等工具绘制动画和3D仿真图,自主研发了毛细管电泳实验的仿真实验软件(电子书)。对仿真实验软件的设计思路、功能模块的实现、教学应用和效果等作了详细叙述。开发的电子书涵盖了实验原理、仪器介绍、样品制备、仪器参数设置、样品测试、数据处理、实验报告等完整的实验流程。电子书的内容完全与我院现行“仪器分析实验”的真实实验和纸质版教材相匹配,从而实现了“虚拟实验”与“真实实验”相融合,线上线下实验教学相融合。电子书中丰富的动画、仿真和实物图片,充分发挥出视觉效应和感观思维的优势,很好地克服了传统实验教学中的时空限制以及纸质版教材单调枯燥的不足,极大地激发学生的学习积极性,提高了学生的学习兴趣和学习效率。

毛细管电泳方法在兽药质量控制中的可行性探讨

兽药质量控制是兽药生产、使用过程中的一个重要环节,通过建立可行的质量标准,对兽药的质量进行有效控制,保障动物源性食品安全。毛细管电泳作为一种以电驱动为主的分离技术手段,近年来得到了广泛的应用。本文将从毛细管电泳发展史、毛细管电泳相关标准、毛细管电泳在化学合成药品、抗生素、中药及其他领域的分析应用等方面,尝试讨论其在兽药行业应用的可行性。

基于毛细管凝胶电泳的寡核苷酸纯度分析方法

在寡核苷酸的化学合成过程中,会产生与全长序列相差一个(n-1)和两个核苷酸(n-2)的杂质,因为分子质量及电荷性质与主成分差异小,难以准确定量检测。毛细管凝胶电泳具有样品用量少、速度快、灵敏度高、分离效果好、自动化程度高等优势,而普遍应用于核酸的分离和分析。采用毛细管凝胶电泳对寡核苷酸纯度进行分析,能有效分离寡核苷酸全长序列与n-1的杂质,在0.03~1.00 mg/L浓度范围内线性良好(R^(2)=0.998 8),检出限约为0.01 mg/L(信噪比S/N>3),该方法的建立将为寡核苷酸的纯度分析与质量控制提供有力的技术支撑。

格隆溴铵立体异构体杂质的毛细管电泳测定——推荐一个药物分析实验

推荐了一个药物分析实验——手性毛细管电泳分析药物格隆溴铵的立体异构体。实验内容包括:环糊精手性选择剂的选择、毛细管电泳分离条件的优化、标准曲线的绘制等。本实验从药物分析的角度,重点介绍毛细管电泳分析方法、手性分离的原理及药物立体异构体杂质分析的重要性。建议在应用化学、药学及制药工程等专业开设。

基于SSR标记的12个非洲菊品种指纹图谱构建及杂交F_(1)后代鉴定

【目的】基于SSR分子标记对12份非洲菊种质进行亲缘关系分析及部分种质杂交F_(1)后代真假杂种鉴定,以期为非洲菊种质资源鉴评及创新利用提供技术支撑。【方法】收集12个非洲菊种质资源,提取其基因组DNA后,利用50对SSR引物进行PCR扩增,通过检测相关多态性结果进行聚类分析和指纹图谱构建,以及部分品种杂交F_(1)后代鉴定。【结果】18对引物在12份非洲菊材料中共扩增出74个多态性等位变异片段,平均每对引物扩增出多态性等位变异片段4.1个。利用74个等位位点,构建了12份非洲菊种质资源的系统进化树,UPGMA聚类分析结果显示,当遗传系数为0.75时,可将受测的12份种质分为5组,Ⅰ组为‘云南红’‘大088’‘情人’,Ⅱ组为‘粉佳人’‘粉球’‘真爱’和‘福娃’,Ⅲ组为‘绣色’‘淑女’和‘黄球’,Ⅳ组和Ⅴ组均只包含1个品种,分别是‘紫水晶’和‘深圳5号’。筛选出GHSSR-1、GHSSR-18、GHSSR-20和GHSSR-21等4对引物,可完全区分12份种质;其中,后3对引物鉴定出兼具双亲特异位点的单株分别是Gh-5、Gh-4和Gh-1,单独1对引物的鉴定率为33%。且受检的3个单株均为真杂种,真杂种率为100%。【结论】成功构建12份非洲菊品种的指纹图谱,综合3对多态性较好的引物鉴定出3株非洲菊杂交F_(1)后代实生苗为真杂种,为非洲菊资源鉴评和创新利用提供可靠的SSR标记引物。

HPV核酸检测及基因分型试剂盒的性能验证及评价

目的对某公司生产的人乳头瘤病毒(HPV)核酸检测及基因分型检测试剂盒(PCR毛细电泳片段分析法)进行性能验证。方法用PCR-反向点杂交技术对待应用的检测HPV亚型试剂盒的准确性、重复性、最低检出限、交叉特异性及抗干扰能力等性能指标进行性能验证及评价。结果该试剂盒采用的PCR毛细电泳片段分析法检测的30例HPV样本中有6例结果不一致,经一代测序验证,与PCR毛细电泳片段分析法检测的结果一致,符合率为100%;重复性和最低检出限均符合要求;解脲支原体、沙眼衣原体、淋球菌和白色念珠菌与该HPV核酸检测及分型试剂盒不存在交叉反应;抗干扰物质对其检测结果无明显影响;与PCR-反向点杂交技术对比,该HPV试剂盒检测对HPV总体阳性率无显著性差异,而对HPV单一感染及多重感染的检出率有差异。结论HPV核酸检测及基因分型试剂盒(PCR毛细电泳片段分析法)性能参数评估合格,可以满足临床样本的检测需求。

The new approach of standardization of capillary electrophoresis

In this paper, we develope the new standardization methods to eliminate the influence in capillary electrophoresis (CE). The markers were used to determine the basis position and then correct the data of sample by the migration time of standard sample, and make the migration time of samples consistent with the standard sample by the criterion of the marker. The problem of time transition was corrected in this way. Then according to the peak height or peak area of the marker in the sample (peak height was used here) compared with the standard sample, the sample data was zoomed appropriately. The absorbance error was made to be correct.The wavelet de-noise method was also used to make the data smooth and get a good baseline.

毛细管电泳法测定青菜中敌百虫的残留量

建立了一种测定青菜中有机磷农药敌百虫的方法,样品以二氯甲烷提取,活性炭脱叶绿素,毛细管电泳法检测,最低检出限为2×10-6mol L,敌百虫浓度在5×10-5～2×10-6mol L之间,呈现良好的线性关系,r=0.9914,样品加标回收率为90%～102%。

毛细管电泳法检测蜂蜜中残留的抗生素

用毛细管电泳法对 5种常用的抗生素四环素 (TC) ,土霉素 (OTC) ,多西环素 (DOC) ,金霉素 (CTC)和氯霉素 (CP)进行了分离。研究了电泳缓冲液的 pH、不同有机试剂添加剂、温度等因素对抗生素分离的影响。实验结果表明 ,体积分数为 4%的N 甲基吗啡啉的加入可大大改善分离效果。在各自相应的浓度范围内 ,峰面积和样品浓度之间呈现良好的线性关系 ,重现性好。氯霉素的检测极限为 10 μg/L ,四环素、土霉素、多西环素为 2 0 μg/L ,金霉素为 40 μg/L(信噪比 >5 )。该方法成功地应用于蜂蜜中残留抗生素的测定 ,具有操作简单、快速方便、灵敏度及自动化程度高、重现性好等优点 ,是对含抗生素类物质进行痕量分析的好方法。