Determination of five isoflavones in Belamcanda Chinensis by RP-HPLC

A reversed phase high performance liquid chromatographic method for the quantitative determination of the five isoflavones (tectoridin, iridin, tectorigenin, irigenin and irisflorentin) in the rhizome of Belamcanda and Iris was developed. The column was packed with TSK gel ODS 80 TS (150 mm x 4.6 mm). Methanol-water (36%-60%) was used as mobile phase. The method was rapid, sensitive, precise and has good reproducibility. The five isoflavones in the crude drugs were completely separated within 40 min. Recoveries of the five constituents were 98.41%-101.93%. The crude drugs were analysed as follows: The powdered sample was refluxed with methanol for 6 hours. The extract was transferred into a 2 ml volumetric flask and diluted to mark with methanol. A definite amount of this sample solution was injected into RP-HPLC. The contents of the five constituents in the rhizomes of Belamcanda and Iris were calculated from the relevant peak heights or peak areas. The analytical results of determination for various species and habitats are shown in Tab 2-5.