Investigating metabolic changes during different organismal or cellular states is of increasing interest. The combination of a data-rich analytical method like mass spectrometry or NMR spectroscopy with a statistical analysis identifies metabolites that are affected by a certain stimulus. Thus, important information on the underlying molecular pathways can be obtained. Here, we describe how to investigate metabolic changes in a model of oncogene-induced senescence. The water-soluble metabolites are isolated by a chloroform-methanol treatment and subsequently analyzed by NMR spectroscopy.