Identification and Quantitative Analysis of Nucleosides and Nucleobases in Aqueous Extracts of Fritillaria Cirrhosa D. Don. Using HPLC–DAD and HPLC-ESI-MS

Abstract

Bulbus Fritillariae cirrhosae is the most widely used medicinal herb for antitussive and apophlegmatic in China and commonly prepared as a water decoction. For clarifying the water-soluble components in Fritillaria cirrhosa D. Don., a simple and sensitive method was developed using high performance liquid chromatography coupled with a photodiode array detector (HPLC-DAD) and an electrospray ionization mass spectrometer (HPLC-ESI/MS) for the quantitative determination and identification of 10 nucleosides and nucleobases. Of the 10 compounds, two (2-deoxyadenosine and hypoxanthine) were identified and determined in F. cirrhosa for the first time. The 10 compounds were separated using a Zorbax SB-Aq column with a gradient elution of methanol and water. All the calibration curves showed good linearity (r2 ≥ 0.9991). The recoveries were between 97.7% and 103.5%. The method was validated with respect to precision, repeatability and accuracy, and was successfully applied to the simultaneous determination of the 10 analytes in 24 batches of F. cirrhosa, collected from different parts of China. This report provided a firm basis for clarifying the pharmacological effect and comprehensively evaluating the quality of F. cirrhosa and related preparations.