To achieve goals in metabolomics investigations, it is necessary to produce a comprehensive
metabolic profiling from biological samples. Identification of metabolites is one of the important steps in
metabolomics studies and the conclusion drawn from such studies depends on how exactly the metabolites
are identified. NMR is one of the most selective analytical techniques which offers structural information
of molecules. But, due to complex biological sample matrix, metabolic identification step needs
application of advanced NMR techniques and analytical strategies for better accuracy. This review covers the analytical
methods and strategies used for identification of metabolites in NMR-based metabolomics. The specific problems and
troubleshoots associated with identification of metabolites in biological samples are discussed in details.
Keywords: NMR spectroscopy, spiking, metabolomics, metabolites assignments, COSY, HSQC.
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