超高效液相色谱-三重四极杆串联质谱法同时测定槟榔籽中四种生物碱含量

郭晓杰; 王盼; 庞朝海; 韩丙军; 阳辛凤

doi:10.13386/j.issn1002-0306.2020090152

超高效液相色谱-三重四极杆串联质谱法同时测定槟榔籽中四种生物碱含量

Simultaneous Determination of Four Alkaloids in Betel Nut Seeds by Ultra Performance Liquid Chromatography-Triple Quadrupole Tandem Mass Spectrometry

摘要

摘要: 目的：采用超高效液相色谱-三重四极杆串联质谱（UPLC-MS/MS）建立槟榔籽中槟榔碱、槟榔次碱、去甲槟榔碱和去甲槟榔次碱含量的检测方法。方法：样品经45%乙醇水溶液提取，利用UPLC-MS/MS法进行检测。色谱条件：采用Waters Atlantis T3（2.1 mm×150 mm，5 μm）色谱柱分离，以0.1%甲酸水溶液-乙腈溶液为流动相进行梯度洗脱，流速0.5 mL·min⁻¹，柱温40 ℃，进样量1 μL。质谱条件：采用电喷雾离子源（ESI⁺），检测方式多反应监测（MRM）模式扫描；外标法定量。结果：在50~500 ng/mL，槟榔碱、槟榔次碱、去甲槟榔碱和去甲槟榔次碱的质量浓度与峰面积都呈现良好的线性关系（R²>0.99）；加样回收率均在规定范围70%~120%内，平均回收率在75.27%~96.70%，RSD<7.0%。结论：该方法的重复性、稳定性良好，仪器精密度较好。

Abstract: Objective: A method was developed for the determination of arecoline, arecaidine, guvacoline and guvacine hydrochloride in areca nut seeds by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Methods: The sample were extracted with 45% ethanol aqueous solution, and detected by UPLC-MS/MS method. Chromatographic conditions: Separated by Waters Atlantis T3 (2.1 mm×150 mm, 5 μm) column, eluted by gradient with 0.1% formic acid aqueous solution-acetonitrile solution as mobile phase, with flow rate of 0.5 mL·min⁻¹, column temperature of 40 ℃, injection volume of 1 μL. Mass spectrometry conditions: Using electrospray ion source (ESI⁺), detection method multiple reaction monitoring (MRM) mode scanning, the quantitative analysis by external standard method. Results: In the range of 50~500 ng/mL, the mass concentration and peak area of arecoline, arecaidine, guvacoline and guvacine hydrochloride showed a good linear relationship (R²>0.99), and the sample recovery rates were all within the specified range of 70%~120%, the average recovery rate was 75.27%~96.70%, and the RSD was less than 7%. Conclusion: The method was good repeatability, stability, and instrument precision.