Huang Dan, Zhao Xinqing, Liu Xiuxiu, Chao Ruobing
West China School of Pharmacy, Sichuan University, Chengdu 610041, P.R. China.
J Chromatogr Sci. 2017 Oct 1;55(9):940-945. doi: 10.1093/chromsci/bmx059.
A convenient and accurate high-performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD) method using solid phase extraction (SPE) was established for quantification of five aminoalcohol-diterpenoid alkaloids (ADAs), including mesaconine, aconine, hypaconine, fuziline and neoline, in the lateral roots of Aconitum carmichaeli (Fuzi) for the first time. The Fuzi extractive was purified using strong cation-exchange SPE. The chromatographic separation was performed on a Gemini C18 column (150 × 4.60 mm, 5 μm) with the mobile phase of methanol-water-diethylamine (48:52:0.01, v/v/v), adjusted to pH 10.2 with acetic acid. The detector was Alltech ELSD 2000ES (drift tube temperature: 90°C; gas flow-rate: 2.3 L/min). Five ADAs in Fuzi were well separated. The calibration curves showed good linearity (r > 0.9990) in the range of 0.0125-0.3750 mg/mL for each alkaloid. The recoveries were in the range of 95.1-105.7%, with relative standard deviations < 5.0%. This method is accurate, specific and repeatable for the determination of five ADAs in different Fuzi samples, which can be applied to the quality control of Fuzi.
首次建立了一种简便、准确的高效液相色谱-蒸发光散射检测法（HPLC-ELSD），该方法采用固相萃取（SPE）对附子侧根中的5种氨基醇二萜生物碱（ADAs）进行定量分析，这5种生物碱包括中乌头碱、乌头碱、次乌头碱、伏毛铁棒锤碱和新乌头碱。附子提取物用强阳离子交换SPE进行纯化。色谱分离在Gemini C18柱（150×4.60 mm，5μm）上进行，流动相为甲醇-水-二乙胺（48:52:0.01，v/v/v），用乙酸调节pH至10.2。检测器为Alltech ELSD 2000ES（漂移管温度：90℃；气体流速：2.3 L/min）。附子中的5种ADAs得到了很好的分离。校准曲线在每种生物碱0.0125 - 0.3750 mg/mL范围内显示出良好的线性（r>0.9990）。回收率在95.1 - 105.7%范围内，相对标准偏差<5.0%。该方法用于测定不同附子样品中的5种ADAs准确、特异且可重复，可应用于附子的质量控制。
