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采用单四极杆质谱法对植物材料中的氨基酸进行高通量分析。

High-throughput analysis of amino acids in plant materials by single quadrupole mass spectrometry.

作者信息

Dahl-Lassen Rasmus, van Hecke Jan, Jørgensen Henning, Bukh Christian, Andersen Birgit, Schjoerring Jan K

机构信息

Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark.

出版信息

Plant Methods. 2018 Jan 19;14:8. doi: 10.1186/s13007-018-0277-8. eCollection 2018.

Abstract

BACKGROUND

The amino acid profile of plants is an important parameter in assessments of their growth potential, resource-use efficiency and/or quality as food and feed. Screening studies may involve large number of samples but the classical amino acid analysis is limited by the fact that it is very time consuming with typical chromatographic run times of 70 min or more.

RESULTS

We have here developed a high-throughput method for analysis of amino acid profiles in plant materials. The method combines classical protein hydrolysis and derivatization with fast separation by UHPLC and detection by a single quadrupole (QDa) mass spectrometer. The chromatographic run time is reduced to 10 min and the precision, accuracy and sensitivity of the method are in line with other recent methods utilizing advanced and more expensive mass spectrometers. The sensitivity of the method is at least a factor 10 better than that of methods relying on detection by fluorescence or UV. It is possible to downscale sample size to 20 mg without compromising reproducibility, which makes the method ideal for analysis of very small sample amounts.

CONCLUSION

The developed method allows high-throughput analysis of amino acid profiles in plant materials. The analysis is robust and accurate as well as compatible with both free amino acids and protein hydrolysates. The QDa detector offers high sensitivity and accuracy, while at the same time being relatively simple to operate and cheap to purchase, thus significantly reducing the overall analytical costs compared to methods based on more advanced mass spectrometers.

摘要

背景

植物的氨基酸谱是评估其生长潜力、资源利用效率和/或作为食物及饲料的质量的一个重要参数。筛选研究可能涉及大量样本,但传统的氨基酸分析受到其耗时极长这一事实的限制,典型的色谱运行时间为70分钟或更长。

结果

我们在此开发了一种用于分析植物材料中氨基酸谱的高通量方法。该方法将传统的蛋白质水解和衍生化与超高效液相色谱(UHPLC)的快速分离以及单四极杆(QDa)质谱仪检测相结合。色谱运行时间缩短至10分钟,该方法的精密度、准确度和灵敏度与其他使用先进且更昂贵质谱仪的近期方法相当。该方法的灵敏度比依赖荧光或紫外检测的方法至少高10倍。在不影响重现性的情况下,可以将样本量缩减至20毫克,这使得该方法非常适合分析极少量的样本。

结论

所开发的方法能够对植物材料中的氨基酸谱进行高通量分析。该分析方法稳健、准确,并且与游离氨基酸和蛋白质水解产物均兼容。QDa检测器具有高灵敏度和准确度,同时操作相对简单且购买成本低廉,因此与基于更先进质谱仪的方法相比,显著降低了总体分析成本。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4de8/5774165/846a48b267b4/13007_2018_277_Fig1_HTML.jpg

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