The Method of Analysis of Hypericin (HPLC) 1.HPLC 1.HPLC condition :0.1mol/lNaH2PO4·2H2O(1.5601g→100ml):=1:4 Mobile phase:0.1mol/lNaH2PO4·2H2O(1.5601g→100ml):Methanol=1:4 :2ml/min Flow Rate:2ml/min :230nm Wave length: 230 nm :0.01Aufs Sensitivity:0.01Aufs :Kromasil C18 Chromatography column:Kromasil C18 :40℃ Column oven temp:40℃ 2. 2.Preparation of Standard solution 0.5mg10ml,. Weigh accurately about 0.5 mg Hypericin standard sample, transfer to a 10 ml volumetric flask. Make up the volume to the mark with Methanol. 3. 3.Preparation of Sample solution 0.1g10ml,20min,,0.45. Weigh accurately about 0.1g Hypericin sample, transfer to a 10 ml volumetric flask. Add appropriate amount Methanol dissolve by sonication. When the temperature of the solution return to the room temperature, make up the volume to the mark with Methanol.Centrifugate the solution,use the Oil system filters to filter supernatant. 4. 4.Calculation %=A1*C*V/(A2*W)*100% Content%=A1*C*V/(A2*W)*100% A1: A1:Sample peak area C: C:Standard sample concentration V: V: Sample dilution A2: A2:Standard Sample peak area W: W:Sample weight
