非衍生化-QuEChERS-超高效液相色谱-串联质谱法快速测定鱼肉中9种生物胺

Rapid determination of 9 kinds of biogenic amines in fish by non-derivatization-QuEChERS-ultra performance liquid chromatography-tandem mass spectrometry

目的建立一种基于Qu ECh ERS前处理技术快速测定鱼肉中9种生物胺(biogenic amines,BAs)含量的超高效液相色谱-串联质谱法,解决衍生化方法操作复杂、条件严格、重现性差等问题。方法样品经乙腈提取、C18和N-丙基乙二胺(primary secondary amine,PSA)净化后,采用Waters HSS T3色谱柱(100 mm×3.0 mm,1.8μm)进行分离,以0.5%甲酸水和0.1%甲酸乙腈为流动相体系进行梯度洗脱,流速为0.3m L/min。采用电喷雾电离(electrospray ionization,ESI)正离子模式,多反应监测(multiple reaction monitoring,MRM)模式检测,标准曲线外标法定量。结果9种生物胺化合物在1.0~100.0μg/L质量浓度范围内线性良好,相关系数(r^(2))在0.9959~0.9998之间,方法检出限(limits of detection,LODs)和定量限(limits of quantitation,LOQs)分别为0.02和0.05 mg/kg。3个浓度水平下,相对标准偏差(relative standard deviations,RSDs)在1.2%~3.6%(n=6)之间,回收率在75.1%~105.6%之间。结论该方法操作便捷、分析时间短、准确度和灵敏度高,适用于鱼肉中9种生物胺化合物的测定。

Objective To develop a method for the rapid determination of 9 kinds of biogenic amines(BAs)in fish based on QuEChERS combined with ultra performance liquid chromatography-tandem mass spectrometry,and solve the problems of complex operation,strict conditions and poor reproducibility of derivations.Methods Samples were extracted with acetonitrile,and purified with C18and primary secondary amine(PSA).The target compounds were separated on a Waters HSS T3 column(100 mm×3.0 mm,1.8μm)by gradient elution using 0.5% formic acid aqueous solution and 0.1% formic acid-acetonitrile as mobile phase at a flow rate of 0.3 mL/min.The detection was performed with electrospray ionization(ESI)in positive mode under multiple reaction monitoring(MRM)mode,the calibration curve was used for the quantification.Results There were good linear relationship for 9 kinds of BAs in the mass concentration range of 1.0-100.0μg/L,their determination coefficients(r^(2))were between 0.9959 and 0.9998.The limits of detection(LODs)and limits of quantitation(LOQs)were 0.02 and 0.05 mg/kg,respectively.The relative standard deviations(RSDs)for 9 kinds of BAs at 3 spiked levels were in the range of 1.2%-3.6%(n=6),the recoveries were in the range of 75.1%-105.6%.Conclusion With simple pretreatment,short analysis time,high accuracy and sensitivity,the method is suitable for the analysis of 9 kinds of BAs in fish.