基质辅助激光解析电离飞行时间质谱法提取阳澄湖全谱图范围内大闸蟹蛋白条件优化

Optimization of extraction conditions of Yangcheng Lake hairy crab protein by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

中文关键词:  基质辅助激光解析电离飞行时间质谱法  阳澄湖大闸蟹  蛋白提取  质谱参数

英文关键词:matrix-assisted laser desorption/ionization time-of-flight mass spectrometry  Yangcheng Lake hairy crab  protein extraction  mass spectrometry parameters

中文摘要:

      目的 建立最佳的蟹肉组织蛋白分析方案, 获得最佳基质辅助激光解析电离飞行时间质谱(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, MALDI-TOF MS)图, 为基于MALDI-TOF MS技术的阳澄湖大闸蟹产地鉴别分析建立条件。方法 从蛋白提取液、基质液以及基质液与提取液配比3个方面对大闸蟹蛋白提取方案进行优化, 从激光强度和聚焦质量2个参数对质谱参数进行优化, 最终获得最佳的阳澄湖大闸蟹蛋白提取及质谱分析方案。结果 通过观察不同条件下的质谱图, 最终选择0.1%三氟乙酸进行蛋白提取, 芥子酸作为基质液, 样品提取液与基质液的上样体积比例选用1:9。而在质谱参数方面, 选择9 μJ的激光强度和10000 Da的聚焦质量。结论 本研究通过比较不同的蛋白提取方法及不同的质谱采集条件, 建立最佳的蟹肉组织蛋白分析方案, 获得最佳质谱图, 为基于MALDI-TOF MS技术的阳澄湖大闸蟹产地鉴别分析建立了条件。

英文摘要:

      Objective To establish the best protein analysis scheme of crab meat tissue, and obtain the best matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) diagram, so as to establish conditions for identification and analysis of hairy crabs in Yangcheng Lake based on MALDI-TOF MS technology. Methods The extraction scheme of hairy crab protein was optimized from 3 aspects of protein extract, matrix solution and ratio of matrix solution to extraction solution, and the parameters of mass spectrometry were optimized from 2 parameters of laser intensity and focusing mass, and finally the optimal scheme of Yangcheng Lake crab protein extraction and mass spectrometry was obtained. Results Through the observation of mass spectra under different conditions, 0.1% trifluoroacetic acid was finally selected to extract the protein, with sinapic acid as the matrix liquid, and the loading volume ratio of sample extract to matrix liquid was 1:9. In terms of mass spectral parameters, the laser intensity of 9 μJ and the focusing mass of 10000 Da were selected. Conclusion In this study, by comparing different protein extraction methods and different mass spectrometry collection conditions, establishe the best protein analysis scheme of crab meat tissue, and obtaine the best mass spectrometry, which establishe the conditions for identifying and analyzing the origin of Yangcheng Lake hairy crabs based on MALDI-TOF MS technology.