Simultaneous determination of eleven constituents in Chenxiang Huazhi Pills by HPLC / 中草药

ABSTRACT

Objective To develop an HPLC method for simultaneous determination of the eleven constituents (agaric-alcohol, naringin, hesperidin, neohesperidin, honokiol, emodin, magnolol, costunolide, dehydrocostus, chrysophanol, and physcion) in Chenxiang Huazhi Pills (CHP) by HPLC with gradient elution. Methods The chromatographic separation was performed on an Thermo Syncronis C18 column (4.6 mm × 250 mm, 5 μm) which was operated at 30 ℃. The mobile phase was a linear gradient prepared from water (A) and acetonitrile (B). The linear gradient elution program was programmed as follows 0—10 min, 20% acetonitrile; 10—20 min, 20%—40% acetonitrile; 20—24 min, 40% acetonitrile; 24—26 min, 40%—52% acetonitrile; 26—30 min, 52% acetonitrile; 30—31 min, 52%—90% acetonitrile; 31—35 min, 90% acetonitrile; 35—40 min, 90%—100% acetonitrile; 40—43 min, 100% acetonitrile; 43—45 min, 100%—20% acetonitrile. The flow rate was 1 mL/min and the detection wavelength was 215 nm. Results The analysis permitted very good separation of eleven constituents within 43 min. A good linear relationship between the peak area and the injection volume was obtained. The ranges of the eleven constituents were 1.4—13.6, 10.0—200.0, 31.5—315.0,1.0—120.1, 1.8—50.6, 0.93—10.1, 1.8—30.0, 0.2—40.3, 1.8—18.1, 1.7—25.0, and 0.45—10.70 μg/mL. The average recoveries of eleven constituents in the samples were in the range of 98.90%—100.87%. The precision RSD of the peak areas of the 11 components ranged from 0.55%—1.54%; Eleven components had good stability within 30 h, and the concentration RSD of each component ranged from 0.75% to 1.94%; The repeatability RSD of each component ranged from 0.39% to 1.73%. The content of agaric-alcohol, naringin, hesperidin, neohesperidin, honokiol, emodin, magnolol, costunolide, dehydrocostus, chrysophanol, and physcion in six batches were 92.0—201.0, 511.5—9 033.0, 5 475.0—12 635.5, 54.5—5 095.5, 192.0—2 137.5, 117.0—391.5, 106.5—1 281.5, 13.0—136.5, 93.5—199.0, 177.0—1 207.0, and 33.5—251.5 μg/g, respectively. Conclusion The method is accurate, rapid and simple with high sensitivity, precision and repeatability, which has been successfully applied as an effective tool for the multicomponent analysis of CHP.