高效液相色谱法-蒸发光散射检测器同时测定肝乐颗粒中黄芪甲苷和芍药苷的含量
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国家自然科学基金项目(81102874)


Simultaneous Determination of Astragaloside and Paeoniflorin in Ganle Granule by High-performance Liquid Chromatography/Evaporative Light-scattering Detector
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    摘要:

    目的 采用高效液相色谱-蒸发光散射检测器(high performance liquid chromatography-evaporative light scattering detector, HPLC-ELSD)建立肝乐颗粒中黄芪甲苷和芍药苷的含量测定方法。方法 以水和甲醇分别为流动相A和流动相B,在Hypersil ODS XB-C18柱(4.6 mm×250 mm,5 μm)上进行梯度洗脱。洗脱程序:0~5 min,A 60%;6~10 min,A 25%,11~18 min,A 10%;流速:1.2 mL/min;柱温:30 ℃。ELSD参数优化:增益值分别设为1、2、3;漂移管温度分别设为50、70、90、110 ℃;载气流量分别为1.2、1.6、2.0、2.4 L/min。结果 通过优化参数,在增益值为2,漂移管温度为90 ℃和载气流量为2.0 L/min时,对芍药苷和黄芪甲苷的含量检测结果最佳。芍药苷和黄芪甲苷的平均加样回收率分别为99.58%、100.37%,RSD分别为2.32%、1.21%。结论 该方法简便、准确、灵敏度高、重现性好,所优化的ELSD参数及建立的HPLC-ELSD法适合肝乐颗粒中芍药苷和黄芪甲苷含量的同时测定。

    Abstract:

    Objective To establish a method for determining the content of astragaloside and paeoniflorin in Ganle Granule by high-performance liquid chromatography/evaporative light-scattering detector (HPLC-ELSD). Methods Astragaloside and paeoniflorin were separated on a Hypersil ODS XB-C18 column (4.6 mm × 250 mm, 5 μm) with mobile phases of water (A) and methanol (B); the gradient elution was performed as follows: 0-5 min, A 60%; 6-10 min, A 25%; 11-18 min, A 10%; the flow rate was 1.2 mL/min and the column temperature was 30 ℃. ELSD parameters were optimized as follows: the gain was set at 1, 2, or 3; the drift tube temperature was set at 50 ℃, 70 ℃, 90 ℃, or 110 ℃; the nitrogen flow rate was set at 1.2 L/min, 1.6 L/min, 2.0 L/min, or 2.4 L/min. Results The optimized conditions for the determination of paeoniflorin and astragaloside were as follows: gain 2; drift tube temperature 90 ℃; nitrogen flow rate 2.0 L/min. The average recovery rates of paeoniflorin and astragaloside were 99.58% (relative standard deviation (RSD) = 2.32%) and 100.37% (RSD = 1.21%), respectively. Conclusion This method is simple, accurate, sensitive, and repeatable. The optimized ELSD parameters and established HPLC-ELSD method are suitable for simultaneous determination of paeoniflorin and astragaloside in Ganle Granule.

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姜 辉,王 斌,高家荣.高效液相色谱法-蒸发光散射检测器同时测定肝乐颗粒中黄芪甲苷和芍药苷的含量[J].安徽中医药大学学报,2015,34(3):86-89

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  • 在线发布日期: 2015-06-04