A simple method is described for the determination of adenosine in royal jelly. The adenosine in the sample was extracted using 80% ethanol and analysed by reversed-phase high-performance liquid chromatography (HPLC). Chromatographic separation was performed using a Dionex HPLC system with a Waters Symmetry C18 column and gradient elution with a mixture of two solvents: solvent A, 0.4% phosphoric acid and solvent B, methanol. The effluent was monitored using a UV detector set at 257nm. The average recoveries were 91.6–98.3% (n=6) with standard deviation below 5.3%. The limits of detection and quantification were 0.017 and 0.048μg/ml, respectively. The method has been successfully applied to the analysis of royal jelly samples. For 45 royal jelly products, the adenosine content varied from 5.9 to 2057.4mg/kg.