超高效液相色谱-串联质谱法测定蜂蜜中18种生物碱及风险评估

Risk Assessment and Determination of 18 Alkaloids in Honey by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry

建立了蜂蜜中马桑内酯、雷公藤类、异喹啉类和吡咯里西啶类等18种生物碱的固相萃取/超高效液相色谱-串联质谱(SPE/UPLC-MS/MS)同时测定方法,并根据实际样品检测结果进行初步风险评估。蜂蜜样品经0.1%甲酸水超声提取、MCX固相萃取柱富集与净化后,使用Waters ACQUITY UPLCTM BEH Phenyl色谱柱(100 mm×2.1 mm,1.7μm)分离,以0.1%甲酸水和甲醇为流动相进行梯度洗脱,采用UPLC-MS/MS进行测定,基质匹配标准曲线法定量。目标化合物在对应质量浓度范围内线性关系良好,相关系数(r^(2))均大于0.993;方法的定量下限为0.05~25μg/kg;平均回收率为77.2%~115%,相对标准偏差(RSD)不大于12%。40份蜂蜜样品中有6份样品检出吡咯里西啶类生物碱,含量为1.09~18.7μg/kg。该方法准确度好,灵敏度高,重现性好,适用于蜂蜜中多种生物碱的同时测定。初步风险评估提示,蜂蜜存在一定的安全风险。

A method for the simultaneous determination of 18 kinds alkaloids including coriaria lactone,tripterygium wilfordii Hook.F,isoquinoline and pyrrolizidine(PAs)in honey samples was proposed by solid phase extraction/ultra performance liquid chromatography-tandem mass spectrometry(SPE/UPLC-MS/MS),and carried out preliminary risk assessment according to the research results.The honey samples were extracted with 0.1%formic water,and subsequently purified and enriched with a mixed solid phase extraction(Oasis MCX SPE)cartridge.Target analytes were separated on a Waters ACQUITY UPLCTM BEH Phenyl column(100 mm×2.1 mm,1.7μm)by gradient mobile phase of 0.1%formic acid water and methanol.Alkaloids was detected under multiple reaction monitoring(MRM)in positive electrospray ionization mode,and quantified using the matrix-matched calibration curves.Good linear relations were obtained with correlation coefficient(r^(2))more than 0.993.The limits of quantitation(LOQs)were in the ranges of 0.05-25μg/kg.The average recoveries for alkaloids in honey samples at three different spiked concentration levels ranged from 77.2%to 115%,with relative standard deviations(RSDs)not more than 12%.The method was used to analyze 40 honey samples,and the PAs were detected in 6 samples with content ranging from 1.09μg/kg to 18.7μg/kg,which indicated to be of potential health risk to human.Due to its advantages of high accuracy and good reproducibility,the proposed method could be applied to the simultaneous analysis of alkaloids in honey samples.