The high-performance liquid chromatographic method using two stationary phases: RP-18 and cyanopropyl silica for the analysis of some alkaloids in Chelidonium maius L. extracts is performed. The extract was chromatographed using a mobile phase composed of an organic modifier (acetonitrile, methanol, tetrahydrofuran or 1,4-dioxan), phosphate buffer and sodium octadecylsulfate or di (2-ethyl hexyl) orthophosphoric acid (HDEHP) as ion-pairing reagents. The influence of the different kind of stationary phase and organic modifiers on retention has been compared. In order to improve the selectivity of separation and peak shape, gradient elution was applied.