发展SPE-LC-MS技术定量分析大肠杆菌胞内能量辅因子

lthough the objective of any 'omic' science is broad measurement of its constitutes, such coverage has been challenging in metabolomics because the metabolome is comprised of a chemically diverse set of small molecules with variable physical propertiesl. Our study aims to establish a robust, sensitive and generic HILIC-HRMS platform to acquire the maximum amount of biological information related to energy cofactors metabolism (ATP, ADP, NAD, NADH, NADP, NADPH) in engineered Escherichia coli cells. To this end, solid phase extraction column (SPE) were selected to remove the interference from the biological matrix or other metabolite. We developed a new LC-MS-based scheme for selected primary metabolite analysis. These metabolites of special biological importance are very diffcult to capture in reported large-scale metabolomic surverys. Targeted approaches for their exteaction, enrichment and LC-MS analysis were developed to accomplish sensitive and selective identification as well as accurate quantitation of these metabolites subset.