代谢组学血浆样品前处理及其快速高分辨液相色谱-质谱分析方法研究  

Plasma Preparation Method for Metabolomics Analysis Based on Rapid Resolution Liquid Chromatography-Mass Spectrometry

作　　者：徐婧[1] 田亚平[1] 陈艳华[1] 张瑞萍[1] 杨芬[2] 宋咏梅[3] 再帕尔·阿不力孜[1] 

机构地区：[1]中国医学科学院,北京协和医学院药物研究所,北京100050 [2]中国医学科学院,北京协和医院,北京100032 [3]中国医学科学院,北京协和医学院肿瘤医院/研究所,北京100021

出　　处：《分析化学》2011年第12期1793-1797,共5页Chinese Journal of Analytical Chemistry

基　　金：国家重大科学研究计划(No.2006CB910600)项目

摘　　要：采用快速高分辨液相色谱-质谱联用技术,对代谢组学研究中血浆样品的前处理方法进行了考察及优化,建立了用于血浆中小分子代谢物分析的前处理方法。通过考察有机溶剂沉淀蛋白(不同溶剂系统)和固相萃取(Solid phase extraction,SPE)两种方法对血浆中蛋白质去除程度及16个代表性化合物的提取效果,确定采用乙腈沉淀蛋白法;进一步对乙腈沉淀蛋白法进行了细致的优化,确定使用3倍体积的冷藏乙腈慢速沉淀可以达到最佳的处理效果。对血浆中10种代表性化合物进行了方法学考察,结果表明,本方法重现性、精密度及样品冻融稳定性均良好。对穿插在检测序列中的生物质控样本检测结果进行主成分分析(Principal componentanalysis,PCA),证明本方法在代谢组学研究中的可重复性及获得的数据的可靠性良好,适用于代谢组学研究。Different methods for plasma preparation in metabolomics analysis were compared in this study.Plasma proteins were either precipitated with four different organic solvents or subjected to solid phase extraction（SPE） on C18-boned phase,with consideration to the number of extracted standard compounds.The eluent was subjected to analyze by rapid resolution liquid chromatography（RRLC）-MS.The results indicated that acetonitrile could effectively precipitated plasma proteins,and had the best extracting effect.Three fold cold acetonitrile coupled with slow precipitation was optimal.The results,according to the reproducibility,precision and stability of the sample,showed that the method was satisfactory and would be applied in metabolomics analysis of plasma preparation.Furthermore,reproducibility of the method verified by PCA analysis is sufficient to ensure data quality in metabolomics applications by carefully monitoring of the quality control samples randomly inserted among the real sample queue.The method could be applied in the metabolomics study.

关 键 词：代谢组学 血浆前处理方法 快速高分辨液相色谱-质谱联用技术 

分 类 号：R927.2[医药卫生—药学] O657.63[理学—分析化学]