title: A facile and sensitive method for quantification of cyclic nucleotide monophosphates in mammalian organs: basal levels of eight cNMPs and identification of 2',3'-cIMP.
abstract: A sensitive, versatile and economical method to extract and quantify cyclic nucleotide monophosphates (cNMPs) using LC-MS/MS, including both 3',5'-cNMPs and 2',3'-cNMPs, in mammalian tissues and cellular systems has been developed. Problems, such as matrix effects from complex biological samples, are addressed and have been optimized. This protocol allows for comparison of multiple cNMPs in the same system and was used to examine the relationship between tissue levels of cNMPs in a panel of rat organs. In addition, the study reports the first identification and quantification of 2',3'-cIMP. The developed method will allow for quantification of cNMPs levels in cells and tissues with varying disease states, which will provide insight into the role(s) and interplay of cNMP signalling pathways.
authors.affiliation: Department of Chemistry, Emory University, Atlanta, GA 30322, USA. xin.jia@emory.edu.
authors.forename: Xin
authors.initials: X
authors.lastname: Jia
authors.affiliation: Department of Chemistry, Emory University, Atlanta, GA 30322, USA. bfontai@emory.edu.
authors.forename: Benjamin M
authors.initials: BM
authors.lastname: Fontaine
authors.affiliation: Department of Chemistry, Emory University, Atlanta, GA 30322, USA. fstrobe@emory.edu.
authors.forename: Fred
authors.initials: F
authors.lastname: Strobel
authors.affiliation: Department of Chemistry, Emory University, Atlanta, GA 30322, USA. emily.weinert@emory.edu.
authors.forename: Emily E
authors.initials: EE
authors.lastname: Weinert
mesh_terms: D000818:Animals; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods; D007289:Cyclic IMP / Q000032:analysis; D057230:Limit of Detection; D008322:Mammals / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells / Q000378:metabolism; D009712:Nucleotides, Cyclic / Q000032:analysis* / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D009712:Nucleotides, Cyclic; D007289:Cyclic IMP