Ion Pair Chromatography for Endogenous Metabolite LC-MS Analysis in Tissue Samples Following HGH Resolution Untargeted Acquisition.

A protocol for the preparation of tissue extracts for the targeted analysis 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 in high-resolution mass spectrometry detection provides highly specific and sensitive acquisition of metabolic intermediates with wide range 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. At last general comments are given to help the assessment of data quality and system performance.