Determination of active component in silymarin by RP-LC and LC/MS

Silybin, isosilybin, silydianin and silycristin in silymarin were separated and quantitatively determined by RP-HPLC. Two diastereoisomers of silybin and isosilybin were respectively separated by RP-HPLC and confirmed by LC/MS. Chromatographic condition consisted of column: Shim-pack VP-ODS (150 x 4.6mm i.d. 5 microm) and Pre-column (10 x 4.6 mm i.d. 5 microm); mobile phase: methanol and solvent mixture (water: dioxane=9:1) by gradient; flow rate: 1.5ml/min; column temp.: 40 degrees C; detector wavelength: 288 nm; The recovery of 99.66% for silycristin, 99.48% for silydianin, 100.0% for silybin and 98.72% for isosilybin was respectively obtained.