摘要
建立了阴离子交换色谱-荧光检测法(AEC-FLD)同时测定人体血清中的酪氨酸(Tyr)、对羟苯基乳酸(PHPLA)和对羟苯基丙酸(PHPA)的分析新方法。样品经乙腈处理沉淀蛋白,离心后取上清液直接进样检测。采用IonPac AG21保护柱(50×2mm)、IonPac AS21分析柱(250×2mm),流动相为4mmol/L NaOH溶液,流速为0.45mL/min,荧光检测器的激发波长和发射波长分别为277nm和340nm。优化条件下三种物质能得到良好的分离和检测。Tyr、PHPLA和PHPA的线性范围分别为0.30-25mg/L、0.025-2mg/L和0.025-2mg/L,相关系数在0.9998-0.9999之间,检出限(S/N=3)分别为0.024mg/L、0.020mg/L和0.019mg/L。三个水平的加标回收率在72.6%-99.1%之间,其相对标准偏差均小于6.62%。该方法简单、快速、准确,可用于实际样品的测定。
A novel,simple,rapid and accurate method was proposed for the simultaneous determination of Tyrosine(Tyr),p-hydroxyphenyllactic acid(PHPLA)and 3-p-hydroxyphenylpropionic acid(PHPA)in human serum by anion exchange chromatography-fluorescence detection(AEC-FLD).The serum samples by acetonitrile deproteinized were injected in AEC-FLD for the separation and detection.Dionex IonPac AS21(250×2 mm)analytical column and IonPac AG21(50×2 mm)guard column were used for the separation of the target analytes.The mobile phase of AEC was 4mmol/L NaOH solution with the flow rate of 0.45mL/min.Fluorescence detector worked at an excitation wavelength of 277 nm and emission wavelength of 340 nm.Under the optimal conditions,Tyr,PHPLA and PHPA were well separated within 20 min.The linear ranges for Tyr,PHPLA and PHPA were 0.30-25mg/L,0.025-2mg/L and 0.025-2mg/L,respectively.The detection limits were 0.024mg/L,0.020mg/L and 0.019mg/L,respectively.At the three spiking levels,the average recoveries were in the range of 72.6%-99.1%,with the relative standard derivations(RSDs)less than6.62%.The results indicate this method is suitable for the real sample analysis.
出处
《分析科学学报》
CAS
CSCD
北大核心
2016年第2期173-177,共5页
Journal of Analytical Science
基金
国家自然科学基金(No.21465017)
江西省自然科学基金(No.20122BAB203013)