Sensitive and simultaneous determination of active components in Lycoris radiata and rat plasma by HPLC with fluorescence detection

Abstract

In this study, a simple, sensitive and rapid reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed for the simultaneous determination of lycorine, lycoramine and galantamine in traditional Chinese medicine Lycoris radiata with fluorescence detection (FD). The chromatographic separation of the three compounds was performed on an Intertsil ODS-SP column. The mobile phase consisted of a mixture of potassium dihydrogen phosphate (KH₂PO₄) solution (15 mM, pH 6.35)–methanol (50 : 50, v/v) delivered at a flow rate of 0.8 mL min⁻¹. The excitation and emission wavelengths for detection were set at 285 and 320 nm, respectively. Under optimum conditions, the baseline separation of these three alkaloids can be carried out within 12 min. The developed method was validated by the limit of detection, limit of quantification, linearity, intra-day precision, inter-day precision and recovery tests. Finally, the developed HPLC-FD method was successfully applied for the simultaneous determination of lycorine, lycoramine and galantamine in the extract of herb Lycoris radiata from different provinces and rat plasma samples after oral administration of the Lycoris radiata extract.