RP-HPLC in determination of seven active ingredients in blood-invigorating and stasi-removing prescription / 第二军医大学学报

ABSTRACT

Objective To develop an RP-HPLC method for determination of tanshinol, protocatechualdehyde, paeoniflorin, puerarin, ferulic acid, tanshinone II A and astragaloside in blood-invigorating and stasis-removing prescription (BSP). Methods The analysiswas performed with a column of Waters Symmetry Shield™ RP C18(150 mm × 4. 6 mm, 3. 5 μm), and the mobile phase consisted of methanol-0. 25% acetic acid. The flow rate was 0. 8 mL/min and the column temperature was 30°C. UV was employed to determine the contents of tanshinol, protocatechualdehyde, paeoniflorin, puerarin, ferulic acid and tanshinone II A, and the detection wavelength was set at 280 nm. Evaporative light scattering detection (ELSD) was employed to determine the contents of astragaloside. The temperature of drift tube was 90°C and the gas flow was 2. 8 L/min (compressed air). Results The linearity was obtained over 0. 01-0. 80 μg (r = 0. 999 8) for tanshinol, 0. 005-0. 4 μg (r=0. 999 7) for protocatechualdehyde, 0. 05-4 μg (r = 0. 999 8) for paeoniflorin, 0. 005-0. 4 μg (r = 0. 999 7) for puerarin, 0.006-0.5 μg (r=1) for ferulic acid, 0. 005-0. 4 μg (r=1) for tanshinone II A, and 0. 031-2. 46 μg (r= 0. 999 3) for astragaloside. The recoverieswere all between 97. 0%-101. 0%, and RSDs were all less than 2%. The contents (mean) of tanshinol, protocatechualdehyde, paeoniflorin, puerarin, ferulic acid, tanshinone H A and astragaloside in three batches of samples were 1. 15, 0. 13, 4. 48, 0. 80, 0. 72, 0. 31 and 3. 12 mg/g, respectively. Conclusion The method in our study is convenient, accurate and sensitive, and it provides a reference for the determination of active ingredients in BSP.