HPLC法同时测定复方抗焦虑胶囊中4种活性成分的含量

目的: 建立HPLC法同时测定复方抗焦虑胶囊中绿原酸、斯皮诺素、橙皮苷、蒙花苷的含量。方法: 采用Agilent ZORBAX Eclipse Plus C₁₈色谱柱(250 mm×4.6 mm,5 μm),以乙腈(A)-0.2%磷酸水(B)为流动相梯度洗脱(0~10 min,11%A→13%A;10~12 min,13%A→16%A;12~27 min,16%A→18%A;27~40 min,18%A;40~60 min,18%A→28%A;60~62 min,28%A→11%A;62~72 min,11%A),流速1 mL·min^-1,检测波长300 nm,柱温30 ℃。结果: 绿原酸、斯皮诺素、橙皮苷、蒙花苷进样量分别在0.076~1.221、0.026~0.410、0.160~2.552和0.007~0.115 μg范围内与峰面积有较好的线性关系,平均加样回收率均在99.8%~101.1%范围内,RSD均小于1.70%。3批样品中绿原酸、斯皮诺素、橙皮苷、蒙花苷含量平均测定结果(n=3)分别为4.14、1.26、7.15、0.31 mg·g^-1。结论: 经方法学验证,本法可用于复方抗焦虑胶囊中绿原酸、斯皮诺素、橙皮苷、蒙花苷的含量测定,为复方抗焦虑胶囊的全面质量控制提供科学依据。

Objective: To simultaneously determine chlorogenic acid, spinosin, hesperidin and buddleoside in anti-anxiety compound prescription capsules by HPLC.Methods: HPLC experiments were performed on Agilent ZORBAX Eclipse Plus C₁₈ column(250 mm×4.6 mm, 5 μm), eluted with acetonitrile(A)-0.1% phosphoric acid(B)as the mobile phases in gradient mode(0-10 min, 11%A→13%A;10-12 min, 13%A→16%A;12-27 min, 16%A→18%A;27-40 min, 18%A;40-60 min, 18%A→28%A;60-62 min, 28%A→11%A;62-72 min, 11%A)at a flow rate of 1 mL·min^-1.The detection wavelength was set at 300 nm.The column temperature was at 30 ℃.Results: The good linearity with peak areas was achieved in the range of 0.076-1.221 μg for chlorogenic acid, 0.026-0.410 μg for spinosin, 0.160-2.552 μg for hesperidin, and 0.007-0.115 μg for buddleoside.The average recoveries were between 99.8% and 101.1% with RSD less than 1.70%.The average contents(n=3)of chlorogenic acid, spinosin, hesperidin and buddleoside were 4.14, 1.26, 7.15 and 0.31 mg·g^-1, respectively.Conclusion: The method can be used in determination of chlorogenic acid, spinosin, hesperidin and buddleoside for the comprehensive quality control by method validation, which can be used for the overall assessment of the quality of anti-anxiety compound capsules.