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Nuclear Magnetic Resonance spectroscopy has emerged as a key tool for understanding metabolic processes in living systems. Recently, a new approach to elucidate metabolism and its mechanisms appeared to the scientific community scene. It is metabonomics: an analysis based on very few assumptions on the biochemical processes that occur in living systems, mainly investigated by advanced spectroscopic techniques including mass spectrometry and NMR spectroscopy. The development of new spectroscopic tools for high thoughput analysis of selected biochemical pathways is crucial for metabolome investigations. The purpose of the present work was to explore the recent NMR improvements by applying and developing new metabolomic strategies for biomarkers discovery, including NMR data handling, peaks quantification and fast data acquisition. This book is integrally extracted from my PhD thesis.