同位素2023，Vol.36Issue(6)：598-604,7.DOI:10.7538/tws.2023.youxian.013
基于液相色谱-同位素稀释质谱法测定氨基酸的蛋白质定量方法
Protein Quantitative Method Based on Determination of Amino Acid by LC-ID-MS
李永利 1高艳秋 1薛民杰 1郝玉红 1李杰1
作者信息
- 1. 上海市计量测试技术研究院,上海市在线检测与控制技术重点实验室,上海 201203
- 折叠
摘要
Abstract
In order to quantitate protein,a liquid chromatography-isotope dilution mass spectrometry method for the analysis of amino acids in protein hydrolysate was estab-lished.The protein was hydrolyzed at 130℃in the presence of 6 mol/L hydrochloric acid solution,and separated by Hypercarb porous graphite carbon chromatography column and 0.1%perfluoropentanoic acid aqueous solution/acetonitrile mobile phase system.Five stable amino acids(proline,valine,leucine,isoleucine and phenylalanine)were selected for quantitative determination with their corresponding stable isotope markers as internal standard,and the absolute quantitative determination of protein was achieved by combining protein sequence information.The linear relationship of the five amino acids is good within the range of 10-1 000 ng/mL(R2＞0.995),the limit of detection and the limit of quantification are between 0.02-0.07 ng/mL and 0.06-0.2 ng/mL,respectively.The recovery at three different levels is between 97.0%-102.4%,and the relative standard deviation is between 1.1%-3.0%.The analysis results of human IgG monoclonal antibody to nucleocapsid protein solution reference material of 2019 Novel Coronavirus and the measurement results of national metrological comparison of 2019-nCovid nucleocapsid protein are satisfactory,which proves that the method has suffi-cient accuracy for protein determination.关键词
液相色谱-串联质谱法/同位素稀释质谱法/氨基酸/蛋白质Key words
LC-MS/MS/isotope diluted mass spectrometry/amino acid/protein分类
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李永利,高艳秋,薛民杰,郝玉红,李杰..基于液相色谱-同位素稀释质谱法测定氨基酸的蛋白质定量方法[J].同位素,2023,36(6):598-604,7.
