超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法快速筛查及测定蜂蜜中20种植物源毒性成分

以超高效液相色谱-四极杆/静电场轨道阱高分辨质谱（ultra-performance liquid chromatography- tandem q-exactive quadrupole-electrostatic field track trap high resolution mass spectrometry，UPLC- Q-Exactive）建立蜂蜜中20种植物源毒性成分的快速筛查和测定方法，基于高分辨质谱的精准分子量和多级碎片信息确立了20种植物源毒性成分的质谱数据库。以纯水提取样品，以混合型阳离子交换固相萃取小柱（Mixed-mode polymeric strong cation exchange and reversed-phase column，MCX）净化，采用AQ-C₁₈色谱柱（2.1 mm×100 mm, 1.9 μm）进行分离，乙腈和含0.1%甲酸的10 mmol/L甲酸铵水溶液作为流动相梯度洗脱，质谱采用全扫描/数据依赖的二级扫描模式（full scan data-dependent MS/MS acquisition mode，Full MS/ddMS2），外标法定量。结果显示，20种化合物在10.0~200.0 μg/L范围内，线性关系良好（r>0.99）；该方法的检出限为0.4~6.9 μg/kg；在3个加标水平下，平均回收率为74.1% ~ 114.6%, 相对标准偏差（RSD）为0.2%~4.7%。该方法灵敏度高，准确可靠，操作简单，可对蜂蜜中20种植物源性毒性成分进行快速筛查和定量分析。

Rapid Screening and Determination of 20 Plant-Derived Toxins in Honey by UPLC-Q-Exactive Quadrupole-Electrostatic Field Track Trap High Resolution Mass Spectrometry

In this study, the method was applied in the rapid screening and determination of 20 plant-derived toxins in honey by ultra-performance liquid chromatography-tandem Q-Exactive quadrupole-electrostatic field track trap high resolution mass spectrometry (UPLC-Q-Exactive). According to precise molecular weight of compounds and multistage fragment ion information detected by UPLC-Q-Exactive, an in-house database of 20 plant-derived toxins was established. The sample was extracted by pure water, and purified on a mixed-mode polymeric strong cation exchange and reversed-phase column (MCX). The extract was separated on an AQ-C₁₈ column (2.1 mm×100 mm, 1.9 μm) through a mobile phase composition of acetonitrile and water (containing 0.1% formic acid and 10 mmol/L ammonium formate) in gradient elution mode. The high resolution mass spectrometry data was simultaneously acquired in full scan data-dependent MS/MS acquisition mode(Full MS/ddMS2), and external standard calibration curves were used for quantification. Results and conclusions were drawn as follows. First, desirable linearities of 20 plant-derived toxins were evaluated among the peak area responses and concentrations in the range of 10.0 to 200.0 μg/L, and the correlative coefficients were over 0.99. The limits of detection were 0.4~6.9 μg/kg. The average recoveries of all the toxins were 74.1%~114.6% at three different levels, and the relative standard deviations (RSDs) ranged between 0.2%~4.7%. This method is sensitive, accurate, reliable and easy to operate, which can be used for rapid screening and quantitative analysis of the 20 plant- derived toxins in honey.