高效液相色谱-串联质谱法测定小鼠肝、肺、肾中Ⅰ、Ⅲ型胶原蛋白含量  

Determination of TypeⅠand TypeⅢCollagen of Liver,Lung and Kidney in Mouse by High Performance Liquid Chromatography-Tandem Mass Spectrometry

作　　者：高建萍[1,2] 张扬 邢芳毓[1] 王俊杰 徐军 孔英俊 罗希[1,2] 张贵锋 GAO Jian-ping;ZHANG Yang;XING Fang-yu;WANG Jun-jie;XU Jun;KONG Ying-jun;LUO Xi;ZHANG Gui-feng(National Key Laboratory of Biochemical Engineering,Institute of Process Engineering,Chinese Academy of Sciences,Beijing 100190,China;School of Chemical and Engineering,University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院过程工程研究所生化工程国家重点实验室,北京100190 [2]中国科学院大学化学工程学院,北京100049

出　　处：《质谱学报》2022年第4期454-462,I0003,共10页Journal of Chinese Mass Spectrometry Society

基　　金：国家重点研发计划(2018YFA0108200,2018YFC1106400);广州市科技计划项目(201803010086);海口市海洋经济创新发展示范城市产业链协调创新类项目(HHCL201804)。

摘　　要：建立了高效液相色谱-串联质谱(HPLC-MS/MS)法测定小鼠肝、肺、肾中Ⅰ、Ⅲ型胶原蛋白含量。首先识别小鼠Ⅰ、Ⅲ型胶原蛋白特征多肽,分别为GSEGPQGVR、GPSGFR。采用子离子及保留时间定性,以GLAGMK为内标对小鼠肝、肺、肾中Ⅰ、Ⅲ型胶原蛋白含量进行分析。结果表明,2个特征多肽在1~500 mg/L浓度范围内的线性关系良好,相关系数均大于0.99,精密度相对标准偏差小于3.7%,加标回收率在86%~118%之间。小鼠生长过程中,肝、肺、肾中Ⅰ型胶原蛋白的比例呈增加趋势,Ⅰ、Ⅲ型胶原蛋白总量呈先增加后降低的趋势。该方法前处理简便高效、精密度高、重现性好,可用于小鼠肝、肺、肾中Ⅰ、Ⅲ型胶原蛋白的精确定量。Collagen is the main component of extracellular matrix,accounting for 25%-30%of total protein.Currently,28 types of collagen have been identified in mammals.The variation of different types of collagen in different tissues is related to age,disease and biological function.TypeⅠandⅢcollagen are the main components in liver,lung and kidney,and their content and ratio are related to inflammation and canceration.Sirius red staining and immunoassay are the traditional method for collagen detection.However,sirius red staining is a semi-quantitative method and immunoassay is not suitable for the determination of collagen content in tissues.In this study,a method of high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)was developed for the determination of typeⅠandⅢcollagen in liver,lung and kidney of mouse.The marker peptides of mouse typeⅠandⅢcollagen were identified and the sequences were GSEGPQGVR and GPSGFR,respectively.The product ions and retention time were used to identify the collagen.The peptide GLAGMK was used as the internal substance to quantitate the content of typeⅠandⅢcollagen in liver,lung and kidney.The results showed that the linear relationship of these two marker peptides was good in the concentration range of 1-500 mg/L,the relative coefficient was more than 0.99,RSD of the assay precision was less than 3.7%,and the recovery of standard addition was between 86%and 118%.The ratios of typeⅠcollagen in liver,lung and kidney were all increased during the growth of mouse.From 0 to 1st week,the ratio of typeⅠcollagen increased rapidly,but they increased slowly from 6th to 12th week.The ratio of typeⅠcollagen was the highest in the kidney,and it was the lowest in the lung.The total content of typeⅠandⅢcollagen increased firstly and then decreased during the growth of mouse.The result was consistent with the trend of total collagen content in liver,lung and kidney,which was measured by hydroxyproline method.The total amount of typeⅠandⅢcollagen was the hi

关 键 词：高效液相色谱-串联质谱(HPLC-MS/MS) 肝 肺 肾 Ⅰ型胶原 Ⅲ型胶原 

分 类 号：O657.63[理学—分析化学]