UHPLC-QTOF-MS/MS-SPE-NMR: A Solution to the Metabolomics Grand Challenge of Higher-Throughput, Confident Metabolite Identifications.
Metabolomics represents a powerful, complementary approach for studying biological system responses to various biotic and abiotic stimuli. A major challenge in metabolomics is the lack of reliable annotations for all metabolites detected in complex MS and/or NMR data. To meet this challenge, we have developed an integrated UHPLC-QTOF-MS/MS-SPE-NMR system for higher-throughput metabolite identifications, which provides advanced biological context and enhances the scientific value of metabolomics data for understanding systems biology. This integrated instrumental method is less labor-intensive and more cost-effective than conventional individual methods (LC; MS; SPE; NMR). It enables the simultaneous purification and identification of primary and secondary metabolites present in biological samples. In this chapter, we describe the configuration and use of UHPLC-MS/MS-SPE-NMR in metabolite analyses ranging from sample extraction to higher-throughput metabolite annotation. With the integrated UHPLC-QTOF-MS/MS-SPE-NMR method, we have purified and confidently identified more than 100 previously known as well as unknown triterpene and flavonoid glycosides while noting that most of the identified compounds are not commercially available.
['Bhatia|Anil|A|', 'Sarma|Saurav J|SJ|', 'Lei|Zhentian|Z|', 'Sumner|Lloyd W|LW|']
D002851:Chromatography, High Pressure Liquid D006801:Humans D009682:Magnetic Resonance Spectroscopy D055442:Metabolome D055432:Metabolomics D052616:Solid Phase Extraction D021241:Spectrometry, Mass, Electrospray Ionization D053719:Tandem Mass Spectrometry
Mass spectrometry (MS) Nuclear magnetic resonance (NMR) spectroscopy Online and offline sample transfer Solid-phase extraction (SPE) Ultrahigh-performance liquid chromatography (UHPLC)