高效液相色谱法测定附子及其炮制品中三种双酯型生物碱

Determination of Three Aconitum Alkaloids in Radix Aconiti Lateralis and Its Prepared Materials by High Performance Liquid Chromatography

本文建立了一种能同时检测附子及其炮制品中乌头碱、新乌头碱和次乌头碱含量的HPLC-DAD检测方法。采用的色谱柱为Hepersil BDS C18分析柱(150 mm×4.6 mm ID,5μm),流动相为40 mmol/L乙酸铵(浓氨水调pH10.0)(A)-乙腈(B)梯度洗脱,检测波长为240nm,流速为1 mL/min。乌头碱、新乌头碱、次乌头碱的线性范围分别为0.0232-2.32μg、0.0216-2.16μg、0.0214-2.14μg(r=0.999976、0.999992、0.999994,n=6);加样回收率为96.4-103.5%(n=6),RSD均小于2.3%。测定结果表明7批不同产地附子生品及16批由同一产地生附子制得的炮制品中三种双酯型生物碱的含量差异悬殊,该方法为附子质量标准和炮制工艺规范的制定及进一步的药效学研究提供了可靠的数据和方法学平台。

By optimizing the extraction, separation and analytical conditions, a reliable and accurate high-performance liquid chromatography （HPLC） method coupled with photodiode array detector （DAD） was developed for simultaneous quantitative determination of aconitine, mesaconitine, hypaconitine in Radix Aconiti Lateralis and its prepared materials. The separation of these Aconitum alkaloids was achieved on a Hypersil BDS C18 column （4.6 mm×150 mm,5μm） with gradient elution using solvents of acetonitrile and ammonium acetate buffer （pH 10.0）. The average recovery rates obtained were in the range of 96.4 103.5% for all with RSDs below 2.3%. The linear ranges of aconitine; mesaconitine; hypaconitine were respectively 0.0232-2.32μg, 0.0216-2.16μg, 0.0214-2.14μg （r=0.999976,0.999992,0.999994,n = 6） O uantitative analysis of the three Aconitum alkaloids in the unprocessed and processed Radix Aconiti Lateralis showed that the contents of the alkaloids varied significantly. This method and quantitation results can provide a scientific and technical platform for setting up a quality control standard of prepared materials of Radix Aconiti Lateralis.