同位素内标超高效液相色谱-串联质谱法检测大鼠血清中丙烯酰胺和环氧丙酰胺

Determination of Acrylamide and Glycidamide in Rat Serum by Ultra Performance Liquid Chromatography Tandem Mass Spectrometry Using Isotope Internal Standards

建立了检测大鼠血清中丙烯酰胺和环氧丙酰胺的超高效液相色谱-串联质谱法(UPLC-MS/MS)。血清样品中加入乙腈沉淀蛋白质,超高速离心取上清液,离心浓缩后,采用ACQUITYUPLCT3 C18色谱柱(50mm×2.1mm,1.7μm),以0.1%甲酸溶液和甲醇为流动相进行梯度洗脱,经UPLC分离,电喷雾离子化四级杆串联质谱以正离子多反应监测方式(MRM)检测,用同位素内标法定量。丙烯酰胺和环氧丙酰胺在0.5～20.0mg/L浓度范围内线性良好,相关系数r均大于0.999;检出限均为0.1mg/L;在3个浓度水平(2.0,8.0,和15.0mg/L)进行添加实验,样品的加标回收率分别为98.0%～108.3%和99.6%～106.7%;日内精密度(RSD)为0.54%～5.8%和1.1%～5.8%;每个样品测试时间仅为6min。本方法测定大鼠血清中的丙烯酰胺和环氧丙酰胺操作简单快速,重现性好,灵敏度高。

A rapid,sensitive and specific method has been developed for the determination of acrylamide and glycidamide in rat serum using ultra performance liquid chromatography tandem mass spectrometry（UPLC-MS/MS） .Serum samples were extracted with acetonitrile,and then centrifugated.Chromatographic separation was achieved on ACQUITY UPLC T3 C18 column （50 mm×2.1 mm,1.7 μm） .The mobile phase consisted of 0.1% aqueous formic acid solution and methanol.After separation with UPLC,acrylamide and glycidamide were detected by electrospray ionization tandem quadrupole mass spectrometry in positive ion multiple reaction monitoring mode（MRM） .Isotope internal standards were used for quantitative analysis.The calibration curves of acrylamide and glycidamide were both linear in the ranges of 0.5-20.0 mg/L.The limits of detection were both 0.1 mg/L for these two analytes.At three spiked levels（2.0,8.0,15.0 mg/L） ,the average recoveries were 98.0%-108.3% with RSD of 0.5%-5.8% for acrylamide and 99.6%-106.7% with RSD of 1.1%-5.8% for glycidamide.Analysis time was only 6 min for each sample.This method was easy to be operated,with good reproducibility and high sensitivity.