基于柱前衍生-超高效液相色谱-质谱联用技术的植物提取液中氨基化合物代谢谱分析

doi:10.3724/SP.J.1123.2015.01028

摘要/Abstract

摘要：

氨基化合物在植物生长发育中起着重要作用。本文建立了一种基于柱前衍生-超高效液相色谱-质谱联用技术的植物提取液中氨基化合物代谢谱的分析方法。以烟草鲜叶为例,共检测出87种氨基化合物。其中43种氨基化合物的定量分析结果表明,方法的线性相关系数在0.993~0.999之间,线性范围可达到4个数量级,检出限为0.03~6.58 ng/mL,日内、日间精密度分别为0.7%~15.6%、0.8%~22.9%,回收率为74.4%~122.7%。采用该方法考察了打顶对不同部位烟草鲜叶中氨基化合物代谢谱的影响,结果显示打顶处理对上部叶影响最大,打顶后上部叶氮代谢主要流向生物碱合成方向。该方法充分利用了三重四极杆串联质谱和高分辨串联质谱技术的优势,可实现植物提取液中氨基代谢物的高选择性、高灵敏度分析。

关键词: 氨基化合物, 超高效液相色谱, 代谢谱, 三重四极杆质谱, 四极杆飞行时间质谱, 植物提取液, 柱前衍生

Abstract:

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.

Key words: amino metabolite, metabolic profiling, plant extract, pre-column derivatization, quadrupole-time of flight mass spectrometry (Q-TOF MS), triple quadrupole mass spectrometry (QQQ MS), ultra high performance liquid chromatography (UHPLC)

中图分类号:

O658

张俊杰, 赵春霞, 赵燕妮, 赵洁妤, 李丽丽, 路鑫, 许国旺. 基于柱前衍生-超高效液相色谱-质谱联用技术的植物提取液中氨基化合物代谢谱分析[J]. 色谱, 2015, 33(6): 613-621.

ZHANG Junjie, ZHAO Chunxia, ZHAO Yanni, ZHAO Jieyu, LI Lili, LU Xin, XU Guowang. Metabolic profiling analysis of amino metabolites in plant extract based on pre-column derivatization-ultra high performance liquid chromatography-mass spectrometry[J]. Chinese Journal of Chromatography, 2015, 33(6): 613-621.

参考文献

[1] Okumoto S, Pilot G. Mol Plant, 2011, 4(3): 453

[2] Zeier J. Plant Cell Environ, 2013, 36(12): 2085

[3] Huang T, Jander G, de Vos M. Phytochemistry, 2011, 72(13): 1531

[4] Ghosh B. Indian J Exp Biol, 2000, 38(11): 1086

[5] Dewey R E, Xie J. Phytochemistry, 2013, 94: 10

[6] Li Y, Wang C H, Zhou C Y, et al. Chinese Tobacco Science (李影, 王程辉, 周初跃, 等. 中国烟草科学), 2013, 34(6): 20

[7] Bouchereau A, Guenot P, Larher F. J Chromatogr B, 2000, 747(1/2): 49

[8] Pena-Gallego A, Hernandez-Orte P, Cacho J, et al. Food Rev Int, 2012, 28(1): 71

[9] Erim F B. TrAC-Trends Anal Chem, 2013, 52(S): 239

[10] Kutlan D, Molnar-Perl I. J Chromatogr A, 2003, 987(1/2): 311

[11] Gomez-Alonso S, Hermosin-Gutierrez I, Garcia-Romero E. J Agric Food Chem, 2007, 55(3): 608

[12] Fiechter G, Sivec G, Mayer H K. J Chromatogr B, 2013, 927(S): 191

[13] Gaudin Z, Cerveau D, Marnet N, et al. Anal Chem, 2014, 86(2): 1138

[14] Sun Y C, Xu X Z, Xu Y L, et al. Chinese Journal of Chromatography (孙言春, 许宪祝, 徐衍岭, 等. 色谱), 2013, 31(3): 275

[15] Li H L, Zhao C X, Zhang J J, et al. Chinese Journal of Chromatography (李好丽, 赵春霞, 张俊杰, 等. 色谱), 2013, 31(12): 1182

[16] Aflaki F, Ghoulipour V, Saemian N, et al. Food Anal Methods, 2014, 7(3): 713

[17] Elbashir A A, Suliman F E O, Aboul-Enein H Y. Appl Spectrosc Rev, 2011, 46(3): 222

[18] Manier M L, Spraggins J M, Reyzer M L, et al. J Mass Spectrom, 2014, 49(8): 665

[19] Mazzotti F, Di Donna L, Napoli A, et al. J Mass Spectrom, 2014, 49(9): 802

[20] Loukou Z, Zotou A. J Chromatogr A, 2003, 996(1/2): 103

[21] Guo K, Li L. Anal Chem, 2009, 81(10): 3919

[22] Gong X, Wang X X, Qi N L, et al. Food Addit Contam, 2014, 31(8): 1431

[23] Yang Y X, Mu C L, Zhang J F. Anal Lett, 2014, 47(8): 1290

[24] Mayer H K, Fiechter G, Fischer E. J Chromatogr A, 2010, 1217(19): 3251

[25] Jacob P, Goniewicz M L, Havel C M, et al. Chem Res Toxicol, 2013, 26(11): 1615

[26] Wang S N, Liu Z, Tang H Z, et al. Microbiol-SGM, 2007, 153: 1556

[27] Yang H Q, Zhou J H, Luo Z M, et al. Hunan Agricultural Sciences (杨虹琦, 周冀衡, 罗泽民, 等. 湖南农业科学), 2004(4): 19

[28] Baldwin I T. J Chem Ecol, 1989, 15(5): 1661

[29] Geng S X, Wang S H, Liu W Q. Chinese Journal of Eco-Agriculture (耿素祥, 王树会, 刘卫群. 中国生态农业学报), 2011, 19(6): 1250

[30] Liu W Q, Cui Z W, Chen X C, et al. Plant Physiology Communications (刘卫群, 崔振伟, 陈旭初, 等. 植物生理学通讯), 2006, 42(6): 1121

[31] Zou Y, Su Y R. Tobacco Science & Technology (邹焱, 苏以荣. 烟草科技), 2008(10): 50

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