Analysis of the main nucleosides in Cordyceps sinensis by LC/ESI-MS

A sensitive, selective and reliable liquid chromatography-mass spectrometry coupled with electrospray ionization interface method for simultaneous separation and determination of thymine, adenine, adenosine and cordycepin in Cordyceps sinensis has been established. The optimum separation for these analytes was achieved using a gradient elution system and a 2.0 x 150 mm Shimadzu VP-ODS column. 2-Chloroadenosine was used as internal standard for this assay. [M+H]+ ions at m/z 127, 136, 268, 252 and 302 were chosen and selective ion monitoring (SIM) mode was used for quantitative analysis of the four main nucleosides. The regression equations were linear in the range of 1.0-117.5 microg x mL(-1) for thymine, 1.8-127.0 microg x mL(-1) for adenine, 0.6-114.0 microg x mL(-1) for adenosine and 0.5-107.5 microg x mL(-1) for cordycepin. The limits of quantitation (LOQ) and detection (LOD) were 1.0 and 0.2 microg x mL(-1) for thymine, 1.8 and 0.6 microg x mL(-1) for adenine, 0.6 and 0.1 microg x mL(-1) for adenosine and 0.5 and 0.1 microg x mL(-1) for cordycepin, respectively. The recoveries of the four nucleosides ranged from 98.47 to 99.32%. The developed method was successfully used to determine nucleosides in Cordyceps sinensis from different sources.