Identification and Quantification of Cyclic Di-Guanosine Monophosphate and Its Linear Metabolites by Reversed-Phase LC-MS/MS

Methods Mol Biol. 2017;1657:45-58. doi: 10.1007/978-1-4939-7240-1_5.


Cyclic dinucleotides such as bis-(3',5')-cyclic dimeric guanosine monophosphate (3',3'-c-di-GMP) represent an important class of second messengers in bacteria and are involved in numerous (patho)physiological settings. Here, we describe a sensitive and specific quantification method for 3',3'-c-di-GMP by HPLC-coupled tandem mass spectrometry (LC-MS/MS). Additionally, linear 3',3'-c-di-GMP metabolites, i.e., 5'-phosphoguanylyl-3',5'-guanosine (pGpG) and 5'-guanosine monophosphate (5'-GMP), as well as cyclic guanosine monophosphate (3',5'-cGMP) and 3',3' c-di-GMP analogues (2',3'-c-di-GMP and 2',2'-c-di-GMP) can be simultaneously determined by this method.

Keywords: Cyclic di-GMP; HPLC; Tandem mass spectrometry.

MeSH terms

  • Bacteria / metabolism
  • Chromatography, Reverse-Phase* / methods
  • Chromatography, Reverse-Phase* / standards
  • Cyclic GMP / analogs & derivatives*
  • Cyclic GMP / chemistry
  • Cyclic GMP / metabolism
  • Metabolomics / methods
  • Second Messenger Systems
  • Tandem Mass Spectrometry* / methods
  • Tandem Mass Spectrometry* / standards


  • bis(3',5')-cyclic diguanylic acid
  • Cyclic GMP