Contemporary investigations regarding the (patho)physiological roles of the non-canonical cyclic nucleoside monophosphates (cNMP) cytidine 3',5'-cyclic monophosphate (cCMP) and uridine 3',5'-cyclic monophosphate (cUMP) have been hampered by the lack of highly specific and sensitive analytic methods for these analytes. In addition, the existence of 2',3'-cNMP besides 3',5'-cNMP has been described recently. HPLC coupled with tandem mass spectrometry (HPLC-MS/MS) is the method of choice for identification and quantification of low-molecular weight endogenous metabolites. In this chapter, recommendations for an HPLC-MS/MS method for 3',5'- and 2',3'-cNMP are summarized.
Keywords: 2′,3′-cNMPs; 3′,5′-cNMPs; Liquid chromatography; Mass spectrometry.