高效液相色谱-串联质谱法同时测定人体尿液中的酪氨酸和色氨酸  

Simultaneous determination of tyrosine and tryptophan in human urine by HPLC-MS/MS

作　　者：支小平[1] 崔盼盼[2] 柳英霞[2] 

机构地区：[1]南昌师范高等专科学校,江西南昌330103 [2]南昌大学分析测试中心,江西南昌330047

出　　处：《南昌大学学报（理科版）》2016年第4期341-345,共5页Journal of Nanchang University(Natural Science)

基　　金：国家自然科学基金资助项目(21465017)

摘　　要：建立了高效液相色谱-串联质谱法(HPLC-MS/MS)同时测定人体尿液中游离酪氨酸和色氨酸分析新方法。采用乙腈-0.1%甲酸溶液(10:90,V:V)为流动相,流速为0.3mL·min^(-1)。在全扫描模式下,将m/z为182.2、205.2的离子分别确定为酪氨酸和色氨酸的母离子,m/z136.1与188.0分别为定量子离子。在质谱的多反应监测正离子模式下进行分析,酪氨酸和色氨酸的检测限分别为0.10和0.25ng·mL^(-1)。在已优化的色谱和质谱条件下,待测样品经过简单的离心、过滤后即可直接进样检测。在3个不同添加水平的回收实验中,酪氨酸和色氨酸的平均回收率为74.0%~124.0%,RSD值小于7.97%,表明该方法可满足实际样品检测要求。该方法操作简单、快速、灵敏度高,可用于人体尿液中游离酪氨酸和色氨酸的同时测定。A new method for the simultaneous determination of free tyrosine and tryptophan in human urine samples by HPLC-MS/MS was developed.The chromatographic separation was achieved on an Agilent XDB-C18 column（4.6×50 mm,1.8μm）,the column temperature was 25 ℃,the mobile phase was CH3CN-0.1%HCOOH（10：90,v/v）,the flow rate was 0.3 mL·min-1 .In the full scan mode,the m/z 182.2 and m/z 205.2 were identified as the precursor ions of tyrosine and tryptophan,the m/z 136.1 and m/z 188 were quantitative product ion,respectively.The analysis was realized by multiple reaction-monitoring modes.Under optimal conditions,the linear ranges of tyrosine and tryptophan were 0.001-2.00μg · mL-1 ,and the limits of detection were 0.10 and 0.25 ng·mL-1 ,respectively.The average recoveries for tyrosine and tryptophan（spiked at three different levels）ranged from 74.0 to 124.0%,the RSD was below 7.97%.The method is simple,rapid and sensitive,and suitable for the simultaneous determination of free tyrosine and tryptophan in human urine.

关 键 词：高效液相色谱-串联质谱 人体尿液 酪氨酸 色氨酸 

分 类 号：O657[理学—分析化学]