title: ¹H-NMR protocol for exometabolome analysis of cultured mammalian cells.
abstract: (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.
authors.affiliation: Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Tiago M
authors.initials: TM
authors.lastname: Duarte
authors.forename: Nuno
authors.initials: N
authors.lastname: Carinhas
authors.forename: Ana Carina
authors.initials: AC
authors.lastname: Silva
authors.forename: Paula M
authors.initials: PM
authors.lastname: Alves
authors.forename: Ana P
authors.initials: AP
authors.lastname: Teixeira
mesh_terms: D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D057230:Limit of Detection; D008322:Mammals / Q000378:metabolism*; D055432:Metabolomics / Q000379:methods*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*