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采用固相萃取-高效液相色谱-蒸发光散射检测法测定附子侧根中5种氨基醇二萜生物碱的含量

Determination of Five Aminoalcohol-diterpenoid Alkaloids in the Lateral Root of Aconitum carmichaeli by HPLC-ELSD with SPE.

作者信息

Huang Dan, Zhao Xinqing, Liu Xiuxiu, Chao Ruobing

机构信息

West China School of Pharmacy, Sichuan University, Chengdu 610041, P.R. China.

出版信息

J Chromatogr Sci. 2017 Oct 1;55(9):940-945. doi: 10.1093/chromsci/bmx059.

Abstract

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.

摘要

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。

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