Jia Xin, Fontaine Benjamin M, Strobel Fred, Weinert Emily E
Department of Chemistry, Emory University, Atlanta, GA 30322, USA.
Biomolecules. 2014 Dec 12;4(4):1070-92. doi: 10.3390/biom4041070.
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.
已开发出一种灵敏、通用且经济的方法，用于使用液相色谱-串联质谱法（LC-MS/MS）在哺乳动物组织和细胞系统中提取和定量环核苷酸单磷酸（cNMPs），包括3',5'-cNMPs和2',3'-cNMPs。解决了复杂生物样品的基质效应等问题并进行了优化。该方案允许在同一系统中比较多种cNMPs，并用于研究一组大鼠器官中cNMPs的组织水平之间的关系。此外，该研究首次鉴定并定量了2',3'-cIMP。所开发的方法将能够定量不同疾病状态下细胞和组织中的cNMPs水平，这将有助于深入了解cNMP信号通路的作用和相互作用。
