1H-NMR Protocol for Exometabolome Analysis of Cultured Mammalian Cells

¹ H-Nuclear magnetic resonance (¹ H-NMR) spectroscopy is a powerful technique to analyze the composition of complex mixtures based on the particular proton fingerprint of each molecule. Here we describe a protocol for exometabolome analysis of mammalian cells using this technique, including sample preparation, spectra acquisition, and integration. The potential of this technique is exemplified by application to cultures of a Chinese hamster ovary (CHO) cell line. The average error associated to this method is under 3% and the limit of quantification for all metabolites analyzed is below 180 μM.