QuEChERS结合高效液相色谱-串联质谱法检测甘草中吡咯里西啶生物碱与风险分析

Determination of pyrrolizidine alkaloids in licorice by QuEChERS combined with high performance liquid chromatography-tandem mass spectrometry and risk analysis

中文关键词:  甘草  吡咯里西啶生物碱  高效液相色谱串联质谱  QuEChERS

英文关键词:licorice  pyrrolizidine alkaloids  high liquid chromatography-tandem mass spectrometry  QuEChERS

中文摘要:

      目的 建立QuEChERS和高效液相色谱-串联质谱法(high performance liquid chromatography-tandem mass spectrometry, HPLC-MS/MS)测定甘草中吡咯里西啶生物碱(pyrrolizidine alkaloids, PAs)的方法。方法 甘草样品经0.05 mol/L硫酸水QuEChERS自动样品制备系统振荡提取后, 用混合型阳离子交换固相萃取小柱(PCX SPE)进行净化, 以5 mmol/L甲酸铵水(含0.1%甲酸)-5 mmol/L甲酸铵甲醇为流动相, Agilent Proshell 120 EC-C18柱进行分离, 利用三重四极杆检测器在多反应监测模式下进行检测。结果 利用建立的前处理方法对甘草中34种PAs做添加回收实验, 在10、20和50 μg/kg 3个浓度下的回收率为71.3%~112.0%, 相对标准偏差为0.29%~8.18%, 满足检测要求。利用建立的提取和检测方法对172个甘草样品中34种PAs进行监测, 阳性样品检出率为22.1%, PAs总含量在9.5~118.0 μg/kg之间。其中石松胺、石松胺N-氧化物、天芥菜碱和大尾摇碱N-氧化物的检出率最高。结论 本研究开发的甘草中PAs的提取和检测方法快速、准确, 可同时测定甘草中34种PAs阳性。所有监测的甘草样品中PAs总量低于欧盟限量标准。

英文摘要:

      Objective To establish a method for the determination of pyrrolizidine alkaloids (PAs) in licorice by QuEChERS combined with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Methods Licorice samples were extracted using 0.05 mol/L sulfuric acid by QuEChERS automatic machine and purified by mixed cation exchange solid phase extraction (PCX SPE). The mobile phase consisted of 5 mmol/L ammonium formate water (containing 0.1% formic acid) and 5 mmol/L ammonium formate methnol. The samples were separated on a Proshell 120 EC-C18 column, and analyzed by triple quadrupole tandem mass spectrometry under the multiple reaction monitoring mode. Results The average recoveries of 34 kinds of PAs in spiked samples ranged from 71.3% to 112.0% at 3 different levels of 10, 20 and 50 μg/kg with the relative standard deviations ranging from 0.29% to 8.18%, meeting the analysis requirements. The 172 licorice samples were analyzed by this method, and 22.1% positive samples were found in those samples with total content in the range of 9.5?118.0 μg/kg. Among all the PAs, lycopsamine, lycopsamine-N-oxide, heliotrine and indicine-N-oxide were the top 4 most frequently found PAs. Conclusion The extraction and detection method for PAs in licorice developed in this study is fast and accurate, and can simultaneously detect 34 kinds of PAs positive in licorice. The total amount of PAs in all monitored licorice samples is below the EU limit standard.