Determination of cyclic guanosine- and cyclic adenosine monophosphate (cGMP and cAMP) in human plasma and animal tissues by solid phase extraction on silica and liquid chromatography-triple quadrupole mass spectrometry

摘要: 3',5'-Cyclic guanosine monophosphate (cGMP) and 3',5'-cyclic adenosine (cAMP) are essential second messenger molecules. They involved in signal transduction within cells, physiological functions such as neurotransmission the modulation of cell growth differentiation organisms, respectively. A quantitative solid phase extraction method (SPE) based on hydrophilic interaction silica was developed applied to both plasma tissue samples. The stable isotope-labeled internal standards ²D₁, ¹⁵N₃-3',5'-cGMP ¹³C₁₀, ¹⁵N₅-3',5'-cAMP were added prior sample preparation ensure high precision accuracy. samples analyzed by reversed-phase liquid chromatography (RP-LC). Negative electrospray (ESI)-MS/MS used selectively monitor several transitions each metabolite. for analysis 3',5'-cAMP 3',5'-cGMP validated range 0.15-20 ng/mL (R²=0.9996 0.9994 3',5'-cGMP, respectively). Basal concentrations fifteen healthy human patients determined with this varied between 4.66-9.20 0.30-1.20 precisions better than 9.1%. together their 2',3'-isomers also a semi way animal tissues. structures isomers confirmed LC-high resolution time-of-flight MS subsequently comparison retention times standards.