高效液相色谱-串联质谱法快速检测植物药酒中的40种生物碱

Rapid determination of 40 kinds of alkaloids in botanical medicated wines by high performance liquid chromatography-tandem mass spectrometry

目的 建立高效液相色谱-串联三重四极杆质谱(high performance liquid chromatography-tandem triple quadrupole mass spectrometry,HPLC-MS/MS)法快速检测植物药酒中40种生物碱的分析方法。方法 样品采用乙腈提取,提取液经N-丙基乙二胺吸附剂净化、氮吹浓缩并以初始流动相溶液定容。待测液经Waters CORTECS C_(18)+色谱柱(100 mm×3.0 mm, 2.7μm)分离,以乙腈-5 mmol/L甲酸铵(含0.1%甲酸)水溶液作为流动相进行梯度洗脱,质谱采用正离子电喷雾离子化、多反应监测模式定性和定量。结果 40种生物碱均具有良好线性关系,检出限为0.03-0.30μg/L,定量限为0.1-1.0μg/L。对植物药酒进行低、中、高质量浓度3水平加标回收试验,回收率范围为60.1%-100.5%,相对标准偏差为1.8%-14.4%(n=6)。结论 本方法样品前处理快速、简单,检测灵敏度高、选择性好,适用于大批量植物药酒样本中多种生物碱的快速分析。

Objective To establish a analysis method for the rapid determination of 40 kinds of alkaloids in botanical medicated wine by high performance liquid chromatography-tandem triple quadrupole mass spectrometry(HPLC-MS/MS). Methods The sample was extracted by acetonitrile, purified by N-propyl ethylenediamine adsorbent,condensed by nitrogen blowing. The residues were redissolved with the initial mobile phase solution. The solution was separated with a Waters CORTECS C_(18)+ column(100 mm×3.0 mm, 2.7 μm) using acetonitrile-5 mmol/L ammonium formate(containing 0.1% formic acid) aqueous solution as mobile phase for gradient elution. Mass spectrometry with electrospray ionization in positive mode and multiple reaction monitoring were used for alkaloids qualitative and quantitative analysis. Results All the 40 kinds of alkaloids had good linear relationship, the limits of detection were 0.03-0.30 μg/L, and the limits of quantification were 0.1-1.0 μg/L. The 3 level standard addition recovery test of low,medium and high concentration was carried out on the botanical wine, and the recoveries were 60.1%-100.5%, and the relative standard deviations were 1.8%-14.4%(n=6). Conclusion The method has the advantages of fast and simple sample pretreatment, high detection sensitivity and good selectivity, and is suitable for the rapid analysis of various alkaloids in large quantities of botanical medicated wine samples.