Abstract:
OBJECTIVE To develop an HPLC method for simultaneous determination of indigo, indirubin and glycyrrhizic acid in Wufu Huadu pill. METHODS An Eclipse XDB-C
18 column(250 mm×4.6 mm, 5 μm) was adopted. The mobile phase was methanol-0.2 mol·L
-1 ammonium acetate-glacial acetic acid(65∶35∶1) at the flow rate of 1.0 mL·min
-1. The UV-detection wavelength was at 250, 288 nm with the column temperature of 25 ℃. RESULTS Calibration curves were found to be linear in the ranges of 2.501-25.01 μg·mL
-1 for indigo, 0.633-6.330 μg·mL
-1 for indirubin and 5.10-51.02 μg·mL
-1 for glycyrrhizic acid. The correlation coefficients were 1.000 0, 0.999 5, 1.000 0. The average recoveries(n=9) of indigo, indirubin and glycyrrhizic acid were 98.3%, 98.4% and 98.2%. CONCLUSION The method is simple, accurate and reproducible. It could be reliable for the quality control of Wufu Huadu Pill.