免疫分析法在体内药物分析中的应用

介绍了放射免疫分析法、酶免疫分析法、化学发光免疫分析法、荧光免疫分析法在体内药物分析中的应用,并比较了免疫分析法和色谱法在体内药物分析中的优缺点.     

常用的中枢神经系统药物体内分析方法

中枢神经系统药物大多具有镇静安眠作用，且有效浓度范围窄，易引起急性中毒或不良反应，故有必要建立有效的体内药物分析方法，对其临床用药进行血药浓度监测。分类综述近年来中枢神经系统药物体内分析方法的研究与发展，其中包括各类色谱法、光谱法及免疫法等。     

色谱新技术在体内药物分析中的应用

综述了国内外用于体内药物分析的一些新兴色谱技术,如色谱-固相微萃取联用法、色谱-质谱联用法、色谱-色谱联用法、超临界流体色谱法、毛细管电泳法、分子生物色谱法、手性色谱法等的具体应用进展;展望了色谱技术在体内药物分析应用中的前景及其发展方向,即开发新的检测技术及借助计算机手段,以实现其连续化、自动化、联用化及智能化.     

传感器技术在体内药物分析中的应用概述

对生物体内肉眼无法观测到的生物活性物质以及药物的分布和代谢进行连续监测,一直是医学家、药理学家和生理学家们研究的目标。传统的体内药物分析方法仅限于色谱法及其与质谱、核磁共振等方法的联用,由于这些方法均需要繁杂的前处理如抽取动物体液后经一系列纯化过程或先处死动物,在组织匀浆后采用复杂的检测设备进行离体检测分析,因此,成本高、耗时长、对机体损害大、不便于进行在体连续监测等。这些缺点使这些方法的应用受到极大的限制。传感器技术可用于在体分析的微型     

限制进入固定相及其在体内药物分析中的应用

在体内药物分析中，样本的制备是最为关键的环节。生物样本成分复杂，除待测药物外，往往含有内源性物质、代谢产物及共存药物等干扰物质。因此，在测定前通常需要花费大量时间进行样本的预处理，如沉淀蛋白、液-液萃取或固相萃取等，冗长繁杂的实验步骤束缚了体内药物分析向快速、自动化方向发展的步伐。     

毛细管电泳在体内药物分析中的应用

介绍毛细管电泳的分离模式、特点及其在血浆、血清、尿液药物分析中的应用,说明它的优越性。指出它在体内药物分析中的广阔应用前景。     

NMR技术在体内药物分析中的应用

ＮＭＲ技术的迅速发展，使其在药学领域中从新药的化学结构的预测扩展到对体液、组织中药物及春代谢物的分析。本文综术字近年发展的几种ＮＭＲ模式在体内药物分析中的应用。     

液相色谱技术在体内药物分析中的应用

现代药学的迅速发展促进针对药物及其代谢物在机体内处置过程的研究不断深入 ,一方面对体内药物分析研究方法和手段提出了越来越高的要求 ,另一方面也推动了体内药物分析研究方法的蓬勃发展。近年来 ,色谱技术特别是液相色谱技术不断发展和完善 ,在灵敏度和选择性等方面都有了很大提高 ,使得对复杂生物样品中药物及其代谢物的测定变得更加准确、快速和简便 ,本文就近年来新型液相色谱技术在体内药物分析中的应用作一简要综述。1　胶束色谱法 (ｍｉｃｅｌｌａｒｃｈｒｏｍａｔｏｇｒａｐｈｙ ,ＭＣ)ＭＣ是一种在常规色谱柱上实现体液样品直接…     

Smart materials for sample preparation in bioanalysis: A green overview

The analysis of biological samples is a complex challenge due to the complexity of the matrix, but also to the low concentration of target analytes that must be determined. Consequently, different sample treatment procedures have been proposed in bioanalysis to clean-up and enrich sample extracts, paying special attention to microextraction approaches. In this frame, the combined use of microextraction approaches with smart materials provides environmentally friendly sample treatment strategies with improved selectivity, sensitivity, and reusability. Applications of smart solid materials includes antibody–antigen interaction based materials, aptamers, molecularly imprinted polymers, metal organic frameworks, restricted access materials, carbon based nanomaterials, and magnetic nanoparticles. On the other hand, smart liquid materials used in sample preparation in bioanalysis comprise bio-based solvents, supramolecular solvents, ionic liquids, and deep eutectic solvents. Synthesis and applications of smart materials have been discussed under a Green Analytical Chemistry perspective.     

The application of fully automated on-line solid phase extraction in bioanalysis

This paper describes the application of fully automated on-line solid phase extraction to the bioanalysis of three example compounds using the Symbiosis platform. The on-line assay performance is compared to off-line methodologies for the same compounds. The three example compounds possess a variety of physicochemical properties and different extraction modes were applied in off-line methods. These methods were developed through optimisation of solid phase or liquid–liquid extraction and chromatographic separation conditions for each of the analytes. Both on-line and off-line methods were evaluated for linearity, carryover, imprecision and inaccuracy. Experiments were also performed investigating modification of ionisation and selectivity against different batches of plasma. On-line and off-line methods were found to be comparable in performance. In conclusion, on-line methodology has distinct advantages for the analysis of large numbers of samples with a marked reduction in manual operation.     

Capillary electrophoresis as a versatile tool for the bioanalysis of drugs--a review

This review article presents an overview of current research on the use of capillary electrophoretic techniques for the analysis of drugs in biological matrices. The principles of capillary electrophoresis and its various separation and detection modes are briefly discussed. Sample pretreatment methods which have been used for clean-up and concentration are discussed. Finally, an extensive overview of bioanalytical applications is presented. The bioanalyses of more than 200 drugs have been summarised, including the applied sample pretreatment methods and the achieved detection limits.     

Inspection by variables as an acceptance criterion in bioanalysis — a proposal

Inspection by variables is proposed as an acceptance criterion for use in bioanalysis. The criteria currently used are deficient either by ignoring the issues of precision (fixed range) and/or accuracy (99% confidence interval), not being able to provide immediate answers (quality charts), or even not being scientifically justified (fixed range). Inspection (sampling) by a variables procedure was originally developed to drive the quality of military supplies (MIL-STD-414) and was consequently incorporated in ISO 3951 as a part of industrial quality control. It is based on the concept of acceptable quality level (AQL), which is the maximum per cent defective (number of results outside of specification per 100 results) that can be considered satisfactory as a process average. It also correlates sample size with batch size. An AQL of 6.5% is proposed as a standard with specification set at ± 20% or ± 15%, which should result in coefficients of variation of approximately 15% and 12%, respectively. This concept has been applied post factum to 14 authentic data sets, thus proving its utility and validity.     

Supercritical fluid chromatography-tandem mass spectrometry for fast bioanalysis of R/S-warfarin in human plasma

Chiral separation for the analysis of enantiomers in biological fluids by HPLC often takes relatively long chromatography time compared to achiral analysis. The advantage of fast mass transfer in packed-column supercritical fluid chromatography (pSFC) and the high-flow compatibility of APCI-MS/MS were applied to develop a fast bioanalytical method for R/S-warfarin in human plasma. Presented here are the main challenges encountered during method development of a semi-automated liquid extraction SFC-MS/MS method. The selection of internal standard, robustness of the SFC equipment, and carryover issues are discussed. The method has high-throughput: the chromatography time is at least two-fold faster than the our fastest previous method; and the liquid/liquid extraction time of 96 samples is less than 20 min using a Tecan Genesis^® RSP 100 pipetting station and a Tomtec Quadra-96^® workstation. The standard curve range was 13.6–2500 ng/ml. Precision of QC concentrations from four validation runs was 7.0% for R-warfarin and 6.0% C.V. for S-warfarin; and the bias was 3.7 and 3.2% R.E., respectively. The method is sensitive, accurate, selective and robust, and was applied to a drug-interaction clinical study with rapid turnaround of sample analysis.     

甲状腺肿瘤患者免疫检验中使用化学发光免疫测定技术的临床价值研究

目的 研究甲状腺肿瘤患者免疫检验中使用化学发光免疫测定技术的临床价值.方法 80例甲状腺肿瘤患者,随机分为实验组和对照组,每组40例.对照组采用放射免疫检测技术,观察组采用化学发光免疫测定技术.对比两组患者检查准确率、平均检测时间.结果 以病理检测结果为金标准,实验组确诊39例,漏诊1例,误诊0例;对照组确诊37例,漏...     

分子印记搅拌棒的研究及应用

搅拌棒吸附萃取技术(SBSE)是从20世纪90年代逐渐发展起来的新型的样品预处理技术,基于固相萃取(SPE)原理,常与高效液相色谱、气相色谱等分析仪器相结合使用,建立的分析方法具有较高的灵敏度以及较好的重现性,应用十分广泛.分子印记技术(MIT)是一项发展迅速的分子识别新技术,制得的分子印记聚合物(MIPs)具有预定性、识别性和广泛的使用性,对模板分子空间结构的"记忆"效应和作用位点的"识别"作用,能够高选择性的分离富集复杂体系中的微量成分.将分子印记技术与搅拌棒吸附萃取技术相结合,弥补了SPE选择性有限的缺点,在复杂样品预处理领域得到了广泛应用.     

Selective solid-phase extraction of cholesterol using molecularly imprinted polymers and its application in different biological samples

Non-covalent molecularly imprinted polymers (MIPs) of cholesterol were prepared by UV initiated polymerization. A polymer that had the highest binding selectivity and capability was used as solid-phase extraction (SPE) sorbents for direct extraction of cholesterol from different biological samples (human serum, cow milk, yolk, shrimp, pork and beef). The extraction conditions of molecularly imprinted SPE (MISPE) were optimized and the optimum protocol was: conditioning MISPE cartridges with n-hexane, loading with n-hexane, washing with n-hexane and n-hexane:toluene = 9:1, respectively, then eluting with chloroform:ethanol:acetic acid = 3:1:1. Cholesterol MISPE selectively recognized, effectively trapped and pre-concentrated cholesterol over a concentration range of 10–80 μg/mL. Recoveries ranged from 80.6% to 92.7%, with R.S.D. lower than 9.8%. Under the optimal condition, MISPE recoveries of spiked human serum, yolk, cow milk, shrimp, pork and beef were 91.1%, 80.4%, 86.6%, 78.2%, 81.4% and 80.1%, respectively. Compared with C18 SPE, almost all of the matrix interferences were removed after MISPE, and better baselines and higher selectivity were achieved.     

A novel strategy for reducing phospholipids-based matrix effect in LC–ESI-MS bioanalysis by means of HybridSPE

A novel strategy to minimize phospholipids-based matrix effects in bioanalytical LC–MS/MS assays was evaluated. The phospholipids-based matrix effect was investigated with a commercially available electrospray ionization (ESI) source coupled with a triple quadrupole mass spectrometer. A systematic comparison approach of two sample preparation procedures was performed. In particular, the matrix effect on mass spectrometry response in rat and human plasma samples was studied by comparing sample extracts obtained by means of a conventional plasma protein precipitation with acetonitrile and the novel HybridSPE-Precipitation procedure.The HybridSPE dramatically reduced the levels of residual phospholipids in biological samples, leading to significant reduction in matrix effects. This new procedure which combines the simplicity of precipitation with the selectivity of SPE allows to obtain much cleaner extracts than with conventional procedures. The effective targeted removal of phospholipids and proteins in biological plasma samples achieved with the HybridSPE-Precipitation procedure provides significant improvement in bioanalysis and a practical and fast way to ensure the avoidance of phospholipids-based matrix effects.     

Regulated LC-MS/MS bioanalysis technology for therapeutic antibodies and Fc-fusion proteins using structure-indicated approach

In recent studies, the development of bioanalysis technologies using liquid chromatography-tandem mass spectrometry (LC-MS/MS) has attracted attention. Our developed nano-surface and molecular-orientation limited (nSMOL) proteolysis enables Fab-specific proteolysis and is optimal for LC-MS/MS analysis of antibody drugs and Fc-fusion proteins in biological samples. In this nSMOL method, antibodies and Fc-fusion proteins are held in pores of the particle and the subsequent proteolysis is carried out with protease-immobilized nanoparticles. The Fab of antibodies or fused region of Fc-fusion protein can be held to orient toward the reaction solution. The access of the immobilized protease is limited to a part in the structure of protein substrate on the particle surface. Thus, nSMOL proteolysis reacts selectively at the Fab complementarity-determining region of antibodies or N-terminal specific domain of Fc-fusion proteins and can be applied to both types of drugs. We have already evaluated drug concentrations in biological samples pretreated with nSMOL proteolysis using LC-MS/MS for more than twenty drugs, of which ten drugs have been fully validated and published. In this review, we discuss the development and application of LC-MS/MS bioanalysis, which enables the bioanalysis of therapeutic antibodies and Fc-fusion proteins by focusing on a structure-based approach.