Ziang Junjie, Zhao Chunxia, Zhao Yanni, Zhao Jieyu, Li Lili, Lu Xin, Xu Guowang
Se Pu. 2015 Jun;33(6):613-21. doi: 10.3724/sp.j.1123.2015.01028.
Amino metabolites are important compounds that play a key role in plant growth and development. A metabolic profiling analysis method of amino metabolites in plant extract was developed based on pre-column derivatization-ultra high performance liquid chromatography- mass spectrometry. Using the tobacco leaf as an example, a total of 87 amino metabolites, including amino acids, amines, peptides, alkaloids etc. were detected. The repeatability of the method was good with RSDs of 85 amino metabolites between 1. 5% and 18. 8%. Forty-three amino metabolites validated by standard samples showed good linearity with the correlation coefficients of 0.993-0.999, covered linear range of four orders of magnitude. The limits of detection were 0.03-6.58 ng/mL. The intra-day and inter-day precisions were 0.7%-15.6% and 0.8%-22.9%, respectively. The recoveries were 74.4%-122.7%. The influence of topping on metabolic profiling of amino metabolites in fresh tobacco was investigated using the developed method. The results showed that the amino metabolites in the upper tobacco leaves were most affected than those in the middle and lower leaves. Metabolism of amino metabolites in the upper leaves after topping was mainly towards the alkaloid synthesis. The method integrated the advantages of triple quadrupole mass spectrometry and high resolution quadrupole-time of flight mass spectrometry. It can be used for metabolic profiling analysis of amino metabolites in plant extract with high sensitivity and selectivity.
氨基酸代谢物是在植物生长发育中起关键作用的重要化合物。基于柱前衍生化-超高效液相色谱-质谱联用技术,建立了一种植物提取物中氨基酸代谢物的代谢谱分析方法。以烟草叶片为例,共检测到87种氨基酸代谢物,包括氨基酸、胺类、肽类、生物碱等。该方法重复性良好,85种氨基酸代谢物的相对标准偏差在1.5%至18.8%之间。经标准样品验证的43种氨基酸代谢物线性良好,相关系数为0.993 - 0.999,线性范围涵盖四个数量级。检测限为0.03 - 6.58 ng/mL。日内和日间精密度分别为0.7% - 15.6%和0.8% - 22.9%。回收率为74.4% - 122.7%。利用所建立的方法研究了打顶对鲜烟叶中氨基酸代谢物代谢谱的影响。结果表明,上部烟叶中的氨基酸代谢物比中部和下部烟叶受影响最大。打顶后上部烟叶中氨基酸代谢物的代谢主要朝着生物碱合成方向进行。该方法综合了三重四极杆质谱和高分辨四极杆-飞行时间质谱的优点。可用于植物提取物中氨基酸代谢物的代谢谱分析,具有高灵敏度和高选择性。