Simultaneous determination of 10 constituents in Boyun Tuiyi Pill by HPLC / 中草药

ABSTRACT

Objective: To develop an HPLC-DAD method for the simultaneous determination of liquiritin, ammonium glycyrrhizinate, vitexicarpin, pulegone, ferulic acid, chlorogenic acid, 3,5-dicaffeoyl quinic acid, berberine hydrochloride, kaempferol, and buddleoside in Boyun Tuiyi Pill (BTP). Methods: Shim-pack VP-ODS C18 column (250 mm × 4.6 mm, 5 μm) was adopted. The mobile phase was composed of methanol-acetonitrile (5050, A) and 0.05% phosphoric acid (B) with gradient elution. Gradient elution 0-5.0 min, 50% A; 5.0-30.0 min, 50%-80% A; 30.0-32.0 min, 80%-50% A; and 32.0-40.0 min, 50% A; Injection volume was 10 μL. The flow rate was 1.0 mL/min and the column temperature was 40℃. Results: liquiritin, ammonium glycyrrhizinate, vitexicarpin, pulegone, ferulic acid, chlorogenic acid, 3,5-dicaffeoyl quinic acid, berberine hydrochloride, kaempferol, and buddleoside were separated well. The linear calibration curves were obtained in 2-20 μg/mL for liquiritin, r = 0.999 2; 20-200 μg/mL for ammonium glycyrrhizinate, r = 0.999 5; 3-30 μg/mL for vitexicarpin, r = 0.999 4; 2-20 μg/mL for pulegone, r = 0.999 7; 1.2-12.0 μg/mL for ferulic acid, r = 0.999 5; 3.5-35.0 μg/mL for chlorogenic acid, r = 0.999 2; 8-80 μg/mL for 3,5-dicaffeoyl quinic acid, r = 0.999 3; 9-90 μg/mL for berberine hydrochloride, r = 0.999 3; 2-20 μg/mL for kaempferol, r = 0.999 6; and 3-30 μg/mL for buddleoside, r = 0.999 5. The average recoveries of the 10 constituents were 99.1%, 101.1%, 100.2%, 99.4%, 101.9%, 98.5%, 100.5%, 101.7%, 100.8%, and 99.7% with RSD of 0.62%, 0.79%, 0.77%, 0.83%, 0.47%, 0.38%, 0.97%, 1.05%, 0.86%, and 1.11%. The contents of six batches of the liquiritin, ammonium glycyrrhizinate, vitexicarpin, pulegone, ferulic acid, chlorogenic acid, 3,5-dicaffeoyl quinic acid, berberine hydrochloride, kaempferol, and buddleoside were 0.505-0.685, 1.793-2.012, 0.227-0.268, 0.183-0.206, 1.258-1.324, 0.348-0.381, 0.648-0.720, 1.544-1.722, 1.543-1.627, and 3.434-3.883 mg/pill, respectively. Conclusion: The method is rapid and has high sensitivity, high accuracy, and good specificity. It can be applied to the quality control of BTP.