Patent ID: 11698361

Abstract:
A method of processing of a biological sample containing multiple metabolites is described The method comprising the steps of pre-treating the biological sample with a metabolite extraction solvent to provide a pre-treated sample, separating a first aliquot of the pretreated sample by reverse phase liquid chromatography (RPLC) to provide a first eluent containing resolved hydrophobic metabolites, and separating a second aliquot of the pre-treated sample by hydrophilic interaction liquid interaction chromatography (HILIC) to provide a second eluent containing resolved hydrophilic metabolites. The first and second eluents are assayed using targeted tandem mass spectroscopy operated in multiple reaction monitoring mode. Each liquid chromatography step (LC) is directly hyphenated with the tandem mass spectrometry (MS/MS) into a single LC-MS/MS analysis. The extraction solvent typically comprises methanol, isopropanol and an acetate buffer.