同位素稀释高效液相色谱-串联质谱法测定乳与乳制品中3-甲氧酪胺和肾上腺素

Determination of 3-Methoxytyramine and Adrenaline in Dairy Products by Isotopic Dilution-Assisted Liquid Chromatography-Tandem Mass Spectrometry

本文建立高效液相色谱-串联质谱法(High performance liquid chromatography-tandem mass spectrometry,HPLC-MS/MS)同时测定乳与乳制品中3-甲氧酪胺和肾上腺素的检测方法。样品前处理使用高氯酸溶液作为提取溶剂,通过弱阳离子固相萃取柱净化,样品采用Waters ACQUITY UPLC BEH C18色谱柱(100 mm×2.1 mm,1.7μm)分离,以0.1%甲酸-甲醇流动相体系进行梯度洗脱,采用电喷雾离子源正离子方式扫描,在多反应监测(Multiple reaction monitoring,MRM)模式下检测,内标法定量。结果表明,3-甲氧酪胺在0.5~12.0 ng/mL,肾上腺素在50~1200 ng/mL范围内线性关系均良好(r>0.99),3-甲氧酪胺的方法检出限为0.2μg/kg,定量限为0.5μg/kg;肾上腺素检出限为20μg/kg,定量限为50μg/kg。在0.5、1.0、3.0μg/kg浓度下,3-甲氧酪胺方法回收率在76.97%~119.20%之间,精密度在1.53%~9.61%之间;在50、100、300μg/kg浓度下,肾上腺素方法回收率在90.65%~108.55%之间,精密度在1.29%~3.95%之间。本方法建立的HPLC-MS/MS结果准确,特异性强,准确度和重复性好,适用于乳与乳制品中3-甲氧酪胺和肾上腺素的分析测定。

A method was established for the simultaneous determination of 3-methoxytyramine and adrenaline in dairy products by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)compled with isotope dilution.Sample was extracted by perchloric acid solution,cleaned by weak cationic solid phase extraction column.The analyte was separated by ACQUITY UPLC BEH C18(100 mm×2.1 mm,1.7μm)column and agradient elution of mobile phase with 0.1%formic acid aqueous solution-methanol.The detection was achieved by HPLC-MS/MS with electrospray ion source and positive ion scanning mode as well as multiple reaction monitoring(MRM)mode.The quantification was accomplished by the internal standard method.It was demonstrated that the linear relationships of 3-methoxytyramine and adrenaline were good in the range of 0.5~12.0 ng/mL and 50~1200 ng/mL(r>0.99),respectively.The detection limits and the limit of quantification of 3-methoxytyramine were 0.2 and 0.5μg/kg,respectively.At the concentrations of 0.5,1.0 and 3.0μg/kg,the recoveries of the 3-methoxytyramine ranged from 76.97%to 119.20%,with the corresponding precision(n=6)of 1.53%~9.61%.While the recoveries of adrenaline at 50,100 and 300μg/kg were 90.65%~108.55%and the relative standard deviations were in the range of 1.29%~3.95%.The method would be accurate in results and good in repeatability,and it can be an important measure to monitor 3-methoxytyramine and adrenaline in dairy products.