Simultaneous determination of 12 chemical components in Tiaojing Pills by RP-HPLC under variable wavelengths / 中草药

ABSTRACT

Objective: To establish a RP-HPLC method to determine the contents of paeoniflorin, acteoside, ferulic acid, leonurine hydrochloride, hesperidin, paeonol, baicalin, asperosaponin VI, limonin, atractylenolide I, atractylenolide III, and pachymic acid in Tiaojing Pills. Methods: The determination was performed on a Venusil MP-C18 column (250 mm × 4.6 mm, 5.0 μm) with ethanol-acetonitrile (40∶60, A) and 0.2% phosphoric acid (B) as mobile phases for gradient elution, at the flow rate of 0.8 mL/min; The column temperature was 45 ℃. Results: The nine components were well separated and showed good linearity, such as paeoniflorin 0.5-50.0 mg/L (r = 0.999 5), acteoside 0.1-10.0 mg/L (r = 0.999 1), ferulic acid 0.2-20.0 mg/L (r = 0.999 2), leonurine hydrochloride 0.3-30.0 mg/L (r = 0.999 3), hesperidin 4.0-400.0 mg/L (r = 0.999 8), paeonol 0.2-20.0 mg/L (r = 0.999 1), baicalin 0.6-60.0 mg/L (r = 0.999 4), asperosaponin VI 1.5-150.0 mg/L (r = 0.999 8), limonin 7.0-700. 0 mg/L (r = 0.999 9), atractylenolide I 0.5-50. 0 mg/L (r = 0.999 3), atractylenolide III 0.5-50. 0 mg/L (r = 0.999 4), and pachymic acid 1.0-100. 0 mg/L (r = 0.999 6). The precision was good, RSD ≤ 0.97%, the repeatability was good in terms of RSD ≤ 1.25% and the recovery rate was 98.5%-103.5% (RSD ≤ 1.24%). Test solution was stable at room temperature within 24 h. The contents of twelve batches of the paeoniflorin, acteoside, ferulic acid, leonurine hydrochloride, hesperidin, paeonol, baicalin, asperosaponin VI, limonin, atractylenolide I, atractylenolide III and pachymic acid were 4.328-4.688, 0.033-0.054, 0.073-0.091, 0.177-0.199, 0.243-0.283, 0.043-0.069, 1.144-1.173, 0.037-0.061, 0.094-0.126, 0.127-0.157, 0.155-0.179, and 0.285-0.327 mg/g, respectively. Conclusion: The method is rapid and has high sensitivity, high accuracy, and good specificity, It can be applied to the quality control of Tiaojing Pills.