液相色谱-串联质谱生物分析方法的基质效应和对策

发布时间：2019-01-02 11:37

【摘要】：液相色谱-串联质谱(LC-MS/MS)法具有高灵敏度、高选择性、高通量等特点,已经成为生物分析的主流方法,广泛应用于新药发现和开发过程中新化学实体及其代谢物的定量分析。然而,由于生物样品基质成分复杂,共洗脱物质会影响分析物的离子化,使分析物的质谱响应增加或降低,从而影响LC-MS/MS分析方法的准确度和精密度。一般而言,离子抑制较离子增强更为常见。因此,在建立LC-MS/MS法时,需要对离子抑制进行评估和校正,并采用不同的策略消除或减少基质效应的影响。本文综述了生物样品分析中基质效应的来源和评估方法,重点介绍了克服基质效应的策略。引起基质效应的物质包括磷脂、盐类、尿素、代谢物等内源性物质和赋形剂、抗凝剂、固定相释放物质及降解产物等外源性物质。目前基质效应的评价方法主要有提取后加入法和柱后灌注法。克服基质效应的策略主要有使用稳定同位素内标,将极性药物衍生化,沉淀蛋白后稀释上清液,优化样品预处理方法、色谱和质谱条件等。结合本课题组的应用实例,重点阐述了建立LC-MS/MS生物分析方法时,为减少基质效应遇到的困难及解决方法。
[Abstract]:Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has become the main method of biological analysis because of its high sensitivity, high selectivity and high throughput. It is widely used in quantitative analysis of new chemical entities and their metabolites in the process of discovery and development of new drugs. However, due to the complex matrix composition of biological samples, co-eluting substances will affect the ionization of the analytes and increase or decrease the mass spectrometric response of the analytes, thus affecting the accuracy and precision of the LC-MS/MS analytical method. In general, ion suppression is more common than ion enhancement. Therefore, when establishing LC-MS/MS method, we need to evaluate and correct the inhibition of ions, and adopt different strategies to eliminate or reduce the effect of matrix effect. In this paper, the source and evaluation methods of matrix effect in biological sample analysis are reviewed, and the strategies to overcome matrix effect are emphasized. The substances that cause matrix effects include endogenous substances such as phospholipids, salts, urea, metabolites and excipients, anticoagulants, stationary release substances, degradation products and other exogenous substances. At present, the evaluation methods of matrix effect mainly include extraction after adding method and post-column perfusion method. The strategies to overcome the matrix effect include the use of stable isotopic internal standards, the derivation of polar drugs, the dilution of supernatants after precipitation of proteins, the optimization of sample pretreatment methods, and the conditions of chromatography and mass spectrometry. Based on the application examples of our group, the difficulties and solutions to reduce matrix effect in the establishment of LC-MS/MS biological analysis method are discussed.
【作者单位】：中国科学院上海药物研究所药物代谢研究中心;
【基金】：国家自然科学基金(81373479,81521005)资助
【分类号】：O657.63

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