液相色谱-同位素稀释质谱法测定血清生长激素  

Determination of Serum Growth Hormone by Liquid Chromatography-Isotope Dilution Mass Spectrometry

作　　者：余利星 翟睿 楚占营 金有训 武利庆 米薇[2] 龚晓云[2] 谢洁 江游[2] 戴新华[2] 方向[2] 俞晓平 YU Li-xing;ZHAI Rui;CHU Zhan-ying;JIN You-xun;WU Li-qing;MI Wei;GONG Xiao-yun;XIE Jie;JIANG You;DAI Xin-hua;FANG Xiang;YU Xiao-ping(Zhejiang Provincial Key Laboratory of Biometrology and Inspection&Quarantine,College of Life Sciences,China Jiliang University,Hangzhou 310018,China;Center for Advanced Measurement Science,Mass Spectrometry Engineering Technology Research Center,National Institute of Metrology,Beijing 100029,China;School of Chemistry and Molecular Engineering,East China University of Science and Technology,Shanghai 200237,China)

机构地区：[1]中国计量大学生命科学学院,浙江省生物计量及检验检疫技术重点实验室,浙江杭州310018 [2]中国计量科学研究院,质谱仪器工程技术研究中心,前沿计量科学中心,北京100029 [3]华东理工大学化学与分子工程学院,上海200237

出　　处：《质谱学报》2021年第2期129-139,共11页Journal of Chinese Mass Spectrometry Society

基　　金：国家重点研发计划项目(2016YFF0200504,2016YFF0200502,2017YFF0106001,2018YFF0212503);国家自然科学基金项目(21575132);重大科学仪器设备开发项目(2016YFF0102603)资助。

摘　　要：复杂基质中低丰度蛋白质的准确定量分析一直是蛋白质研究的重点和难点。随着质谱技术的飞速发展,同位素稀释质谱法成为血清中低丰度蛋白质准确定量分析的重要方法。本研究以同位素标记的人生长激素为内标,通过C12和C4色谱柱两次分离收集目标馏分,建立了基于离线二维高效液相色谱分离血清样本的新方法。然后,结合高效液相色谱-同位素稀释质谱法,准确定量了人血清低丰度蛋白质生长激素的含量。通过对模拟样品(空白血清添加已知质量生长激素标准物质,理论含量为12.00 ng/g)和国际比对样品中生长激素(浓度未知)的测量及不确定度计算,定量结果分别为(11.45±2.33)ng/g和(12.84±1.46)ng/g。该方法准确性高、重复性好,为复杂基质低丰度蛋白质的准确定量分析提供了一种有效的测量方法和可溯源的分析结果。The accurate and quantitative analysis of low-abundance proteins in complex matrices has always been the focus and faced difficulty in protein research.With the rapid development of mass spectrometry technology,isotope dilution mass spectrometry has become an important method for accurate quantitative analysis of low-abundance proteins in serum.In this study,isotopically labeled human growth hormone was used as an internal standard.A strategy for separation of serum samples was established based on off-line two-dimensional high-performance liquid chromatography(2D HPLC).Then,combined with high performance liquid chromatography-isotope dilution mass spectrometry(IDMS)method,quantitative analysis of low-abundance protein growth hormone(hGH)in human serum was performed with high accuracy.Based on the measurement and uncertainty calculation of the simulated samples(blank serum with known quality of growth hormone standard substance,theoretical concentration 12.00 ng/g)and international comparison samples,the quantitative results are(11.45±2.33)ng/g and(12.84±1.46)ng/g,respectively.This method has high accuracy and good repeatability,and can provide an effective measurement method for accurate quantitative analysis of low-abundance proteins in complex matrices.The results can trace to reference material.

关 键 词：血清 生长激素 高效液相色谱(HPLC) 同位素稀释质谱法(IDMS) 定量分析 

分 类 号：O657.63[理学—分析化学]