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
Xin JiaDepartment of Chemistry, Emory University, Atlanta, GA 30322, USA
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Biomolecules 2014, 4(4), 1070-1092; https://doi.org/10.3390/biom4041070
Received: 8 August 2014 / Revised: 27 November 2014 / Accepted: 1 December 2014 / Published: 12 December 2014
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.
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Keywords:
liquid chromatography; tandem mass spectrometry; LC-MS/MS; 3',5'-cyclic nucleotides monophosphate; 2',3'-cyclic nucleotides monophosphate; inosine 2',3'-cyclic monophosphate; mammalian cNMP basal levels
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited
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MDPI and ACS Style
Jia, X.; Fontaine, B.M.; Strobel, F.; Weinert, E.E. 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. Biomolecules 2014, 4, 1070-1092.
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