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Separation and identification of the constituents in Fangchi Radix of different origins

Fangchi Radix is derived from Aristolochiaceae or Menispermaceae plants and contains a series of biologically active alkaloids. Fifteen alkaloids in Fangchi crude extracts, including acutumidine (1), magnoflorine (2), stepharine (3), sinomenine (4), acutumine (5), cyclanoline (6), fangchinoline (7), berbamine (8), tetrandrine (9), isotetrandrine (10), trilobine (11), aristolochic acid Π (12), aristolochic acid Ι (13), aristololactam (14) and isotrilobine (15), were separated completely by high-performance liquid chromatography (HPLC) within 60 min. UV detection at 254 nm with a linear gradient elution consisting of 20 mM CH3COONH4 (pH 4.81) and H2O-CH3CN was found to be the most efficient eluent for the LC and LC-MS separations. The structures of the compounds corresponding to the peaks in the LC chromatogram were identified by LC-MS. A total of 32 commercial samples of Fangchi Radix from Sinomenium acutumn, Stephania tetrandra, Aristolochia fangchi and Cocculus trilobus were collected from herbal markets in Taiwan, Japan and China. The contents (peak-area ratios) and structures of the fifteen constituents in these samples were readily determined by the proposed methods and the results could be used to estimate the origin and quality of an herbal drug.