超高效液相色谱-四极杆-飞行时间质谱法测定牛奶中51种激素

Determination of 51 Hormones in Milk by Ultrahigh Performance Liq⁃uid Chromatography-Quadrupole-Time of Flight Mass Spectrometry

基于超高效液相色谱-四极杆-飞行时间质谱(UPLC-Q-TOF MS),建立了牛奶中51种激素的高通量筛选和定量方法,并采用信息依赖型采集(IDA)模式建立了51种激素的定性筛选数据库。牛奶样品经乙腈沉淀蛋白后,采用氧化锆包覆硅胶(Z-Sep)填料进一步净化。采用Waters Acquity BEH C_(18)(100 mm×2.1 mm,1.7µm)色谱柱进行分离,UPLC-Q-TOF MS测定,基质匹配曲线外标法定量。牛奶中51种激素的线性范围为0.10~500µg/L,相关系数(r^(2))均大于0.99;检出限为0.03~1.67µg/kg,定量下限为0.10~5.00µg/kg;在5、10、50µg/kg加标水平下的回收率为44.6%~120%,相对标准偏差为0.90%~20%。利用该方法测定了20份市售牛奶样品,其中19份样品检出孕酮,检出量为0.80~4.51µg/kg;1份样品中含有痕量的氢化可的松,检出量为0.62µg/kg;其他激素均未检出。该方法具有良好的线性、灵敏度、准确度和精密度,可用于多种激素的同时定性定量分析。

A comprehensive high-throughput screening and quantification method for hormones was established based on ultrahigh performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF MS).A database for qualitative screening of 51 hormones was estab-lished using information dependent acquisition(IDA)mode.After precipitation of protein in milk samples by acetonitrile,the milk samples were purified using zirconia coated silica gel sorbent(Z-Sep).Separation was performed on Waters Acquity BEH C_(18) column(100 mm×2.1 mm,1.7µm)with gradient elution by UPLC-Q-TOF MS and quantified using matrix matching curve external stan-dard method.The method showed good linearity in the range of 0.10-500µg/L,with correlation co-efficients(r^(2))greater than 0.99.The limits of detection(LODs)and limits of quantitation(LOQs)were in the range of 0.03-1.67µg/kg and 0.10-5.00µg/kg,respectively.The recoveries is 44.6%-120%with relative standard deviations of 0.90%-20%.Twenty milk samples were determined using this method,of which 19 samples were detected for progesterone,with content ranged from 0.80 to 4.51µg/kg,one sample had trace amounts of cortisol detected at 0.62µg/kg,and no other hormones were detected.This method has good linearity,sensitivity,accuracy,and precision,and can beused for simultaneous qualitative and quantitative analysis of multiple hormones.