HPLC-CAD法测定小柴胡片中琥珀酸、盐酸麻黄碱、黄芩苷、汉黄芩苷、柴胡皂苷a、柴胡皂苷d、甘草酸、甘草次酸和棕榈酸

Determination of succinic acid,ephedrine hydrochloride,baicalin,wogonoside,saikosaponin a,saikosaponin d,glycyrrhizic acid,glycyrrhetinic acid,and palmitic acid in Xiaochaihu Tablets by HPLC-CAD

目的 建立高效液相色谱联合电喷雾检测器(HPLC-CAD)测定小柴胡片中琥珀酸、盐酸麻黄碱、黄芩苷、汉黄芩苷、柴胡皂苷a、柴胡皂苷d、甘草酸、甘草次酸、棕榈酸的分析方法。方法 采用GL Science Inertsil ODS-3色谱柱(250mm×4.6 mm,3μm);流动相:5 mmol/L甲酸铵(甲酸调pH值至4.2)–乙腈,梯度洗脱;柱温:35℃;进样量:20μL;体积流量:0.7 mL/min;CAD雾化器温度为35℃,气压为61.0 psi,采集频率为10 Hz。结果 琥珀酸、盐酸麻黄碱、黄芩苷、汉黄芩苷、柴胡皂苷a、柴胡皂苷d、甘草酸、甘草次酸、棕榈酸的线性范围分别为0.94~18.74、1.23~24.66、8.90~178.06、3.51~70.20、1.52~30.36、1.25~25.02、3.48~69.54、2.50~50.04、1.23~24.56μg/mL;平均回收率分别为97.05%、97.82%、97.85%、96.46%、98.53%、97.58%、97.05%、97.65%、97.03%,RSD值分别为1.46%、2.16%、0.83%、1.00%、1.74%、1.49%、0.75%、1.68%、1.46%。结论 本法简便、准确,为小柴胡片的质量标准提升研究提供有效的技术手段。

Objective To develop an HPLC-CAD method for simultaneous determination of succinic acid, ephedrine hydrochloride, baicalin, wogonoside, saikosaponin a, saikosaponin d, glycyrrhizic acid, glycyrrhetinic acid, and palmitic acid in Xiaochaihu Tablets. Methods The chromatographic separation was performed on a GL Science Inertsil ODS-3 column GL(250 mm × 4.6 mm, 3 μm) with 5 mmol/L ammonium formate(formic acid ajust pH 4.2)-acetonitrile as the mobile phase with gradient elution. The column temperature was set at 35 ℃, injection volume was 20 μL, and flow rate was 0.7 mL/min. For CAD, the nebulization temperature of detector was set as 35 ℃, the gas pressure was 61.0 psi, and acquisition frequency was 10 Hz. Results The linear ranges of succinic acid, ephedrine hydrochloride, baicalin, wogonoside, saikosaponin a, saikosaponin d, glycyrrhizic acid, glycyrrhetinic acid, and palmitic acid were 0.94 — 18.74, 1.23 — 24.66, 8.90 — 178.06, 3.51 — 70.20, 1.52 — 30.36, 1.25 — 25.02, 3.48 — 69.54, 2.50 — 50.04, and 1.23 — 24.56 μg/mL, respectively. The average recoveries were 97.05%, 97.82%, 97.85%, 96.46%, 98.53%, 97.58%, 97.05%, 97.65%, and 97.03% with RSD values of 1.46%, 2.16%, 0.83%, 1.00%, 1.74%, 1.49%, 0.75%, 1.68%, and 1.46%, respectively. Conclusion The method is simple and accurate, which can provide an effective technical means for the study on the improvement of the quality standard of Xiaochaihu Tablets.