靶向采集后组织样本中内源性代谢物LC-MS分析的离子对色谱法

Ion Pair Chromatography for Endogenous Metabolites LC-MS Analysis in Tissue Samples Following Targeted Acquisition.

作者信息

Michopoulos Filippos

机构信息

IMED Oncology, AstraZeneca, Macclesfield, Cheshire, UK.

出版信息

Methods Mol Biol. 2018;1738:83-97. doi: 10.1007/978-1-4939-7643-0_6.

DOI:10.1007/978-1-4939-7643-0_6

PMID:29654584

Abstract

A protocol for the preparation of tissue extracts for the targeted analysis of ca. 150 polar metabolites, including those involved in central carbon metabolism is described, using a reversed-phase ion pair U(H)PLC-MS method. Data collection enabled by multiple-reaction monitoring provides highly specific, sensitive acquisition of metabolic intermediates with a wide range of physicochemical properties and pathway coverage. Technical aspects are discussed for method transfer along with the basic principles of sample sequence setup, data analysis, and validation. General comments are given to help the assessment of data quality and system performance.

摘要

本文描述了一种使用反相离子对U(H)PLC-MS方法制备组织提取物的方案，用于对约150种极性代谢物进行靶向分析，包括那些参与中心碳代谢的代谢物。通过多反应监测实现的数据采集能够高度特异性、灵敏地获取具有广泛物理化学性质和途径覆盖范围的代谢中间体。讨论了方法转移的技术方面以及样品序列设置、数据分析和验证的基本原理。给出了一般性评论以帮助评估数据质量和系统性能。