纳米金标记液相色谱-电感耦合等离子体质谱法测定人血清中转铁蛋白  

Quantification of Human Serum Transferrin Based on High Performance Liquid Chromatography-Inductively Coupled Plasma-Mass Spectrometry with Gold Nanoparticles Labeling

作　　者：韩亚琛 冯流星[2] 李红梅[2] 熊金平[1] HAN Ya-Chen;FENG Liu-Xing;LI Hong-Mei;XIONG Jin-Ping(College of Material Science and Technology,Beijing University of Chemical Technology,Beijing 100029,China;National Institute of Metrology,Beijing 100029,China)

机构地区：[1]北京化工大学,材料科学与工程学院,北京100029 [2]中国计量科学研究院,北京100029

出　　处：《分析化学》2020年第2期206-214,共9页Chinese Journal of Analytical Chemistry

基　　金：国家重点研发计划项目(No.2017YFF0205402);国家自然科学基金项目(No.11475163)资助~~

摘　　要：建立了基于纳米金(AuNPs)标记并结合高效液相色谱(HPLC)-电感耦合等离子质谱(ICP-MS)的联用技术,通过测量转铁蛋白(Tf)上标记的AuNPs的量,实现了标准Tf及人血清中Tf的准确定量分析。在本研究中,将Tf与过量的AuNPs在磷酸盐缓冲液(pH 6.8)中孵育,形成AuNPs-Tf标记物,之后通过HPLC分离AuNPs-Tf,标记后Tf分子上附着的AuNPs上的Au原子可起到增强信号的作用,通过ICP-MS在线检测AuNPs-Tf上Au的信号,实现复杂血清样品中Tf的准确定量分析。本方法中Tf浓度与AuNPs-Tf上的Au信号强度之间线性关系良好(R 2=0.9959),线性范围宽(3个数量级),方法检出限低至6 ng/mL。本方法经标准Tf验证后,用于人血清标准物质(ERM-DA470/IFCC)中Tf的检测,测量结果在标准物质不确定度范围内,回收率在95.2%~102.6%之间。由于AuNPs的信号增强效应和ICP-MS的灵敏检测,相比于传统的以吸光度为检测方式的酶联免疫法,本方法在检出限和灵敏度方面均有明显提升。A simple and sensitive method for quantification of transferrin(Tf)in human serum by high performance liquid chromatography(HPLC)coupled with inductively coupled plasma mass spectrometer(ICP-MS)with gold nanoparticles(AuNPs)labeling was established.Accurate quantification of standard Tf and Tf in human serum were achieved by measuring the concentration of AuNPs labeled on Tf.Tf could be incubated with most amount of AuNPs to form AuNPs-Tf in phosphate buffer(pH 6.8).Since gold atoms labeled to the Tf molecule could enhance the signals,accurate quantification of Tf in human serum could be achieved by gold signal measurement with ICP-MS after AuNPs-Tf was separated by HPLC.Good linear correlation coefficient(R 2=0.9959)was obtained between Au peak area on AuNPs-Tf and concentration of Tf with a wide linear response over 3 orders of magnitude,and the limit of detection was 6 ng/mL.The developed method was successfully applied for detection of Tf in human serum certified reference material(ERM-DA470/IFCC)after being verified by standard Tf.Good agreement was achieved and the quantitative recovery was in the range of 95.2%-102.6%.Due to signal amplification of AuNPs and ultra-sensitive detection of ICP-MS,the sensitivity and accuracy were improved significantly compared to traditional enzyme-linked immunosorbent assay.Moreover,the measurement results could be directly traced to SI unit.

关 键 词：高效液相色谱-电感耦合等离子质谱 纳米金 转铁蛋白 

分 类 号：R446.1[医药卫生—诊断学] O657.63[医药卫生—临床医学]