HPLC法同时测定长喙乌头中3种主要二萜类生物碱含量

目的: 建立HPLC法同时测定长喙乌头Aconitum georgei Comber.中滇乌碱、草乌甲素和黑乌弱碱的含量。方法: 采用Agilent extend-C₁₈色谱柱(250 mm×4.6 mm,5 μm),以乙腈-40 mmol·L^-1乙酸铵水溶液(氨水调pH至9.5)为流动相梯度洗脱,流速1 mL·min^-1,柱温30 ℃;检测波长260 nm。结果: 滇乌碱、草乌甲素和黑乌弱碱质量浓度分别在0.14~1 400 μg·mL^-1(r=1.000 0)、0.086 8~868 μg·mL^-1(r=1.000 0)、0.064~128 μg·mL^-1(r=1.000 0)范围内内呈良好的线性关系,平均加样回收率分别为102.7%、102.1%、105.8%,RSD分别为2.5%、1.3%、2.2%。4批长喙乌头样品中滇乌碱、草乌甲素、黑乌弱碱的质量分数测定结果分别为62.00~1 037.65、386.99~2 448.99、11.17~96.00 μg·g^-1。结论: 经方法学验证,本法可用于同时测定长喙乌头中的滇乌碱、草乌甲素和黑乌弱碱的含量,为该药的质量控制和临床用药安全提供参考。

Objective: To establish an HPLC method for simultaneous determination of yunaconitine, bulleyaconitine A and foresaconitine in the roots of Aconitum georgei.Methods: The determination method was developed on an Agilent extend-C₁₈ column(250 mm×4.6 mm, 5 μm).The gradient elution was carried out with acetonitrile-40 mmol·L^-1 ammonium acetate(adjusted to pH 9.5 with ammonium hydroxide)as mobile phase at a flow rate of 1 mL·min^-1.The column temperature was 30 ℃ and detection wavelength was 260 nm.Results: Yunaconitine, bulleyaconitine A and foresaconitine each showed a good linearity in the range of 0.14-1 400 μg·mL^-1(r=1.000 0), 0.086 8-868 μg·mL^-1(r=1.000 0), and 0.064-128 μg·mL^-1(r=1.000 0)respectively;and the average recoveries were 102.7%(RSD=2.5%), 102.1%(RSD=1.3%), and 105.8%(RSD=2.2%), respectively.The contents in 4 samples of A.georgei were 62.00-1 037.65 μg·g^-1 for yunaconitine, 386.99-2 448.99 μg·g^-1 for bulleyaconitine A, and 11.17-96.00 μg·g^-1 for foresaconitine.Conclusion: According to the methodology validation, the established method can be applied to simultaneous determination of yunaconitine, bulleyaconitine A and foresaconitine in roots of A.georgei, which could be used for the quality and safety control of this species.