An optimized method for NMR-based plant seed metabolomic analysis with maximized polar metabolite extraction efficiency, signal-to-noise ratio, and chemical shift consistency

CORC > 武汉物理与数学研究所 > 中国科学院武汉物理与数学研究所 > 磁共振应用研究部

	An optimized method for NMR-based plant seed metabolomic analysis with maximized polar metabolite extraction efficiency, signal-to-noise ratio, and chemical shift consistency
	Wu, Xiangyu 1; Li, Ning 1; Li, Hongde 1; Tang, Huiru 1,2
刊名	ANALYST
	2014
卷号	139 期号:7 页码:1769-1778
英文摘要	Plant metabolomic analysis has become an essential part of functional genomics and systems biology and requires effective extraction of both primary and secondary metabolites from plant cells. To establish an optimized extraction method for the NMR-based analysis, we used the seeds of mungbean (Vigna radiata cv. Elu no. 1) as a model and systematically investigated the dependence of the metabolite composition in plant extracts on various extraction parameters including cell-breaking methods, extraction solvents, number of extraction repeats, tissue-to-solvent ratio, and extract-to-buffer ratio (for final NMR analysis). We also compared two NMR approaches for quantitative metabolomic analysis from completely relaxed spectra directly and from partially relaxed spectra calculated with T-1. By maximizing the extraction efficiency and signal-to-noise ratio but minimizing inter-sample chemical-shift variations and metabolite degradations, we established a parameter-optimized protocol for NMR-based plant seed metabolomic analysis. We concluded that aqueous methanol was the best extraction solvent with an optimal tissue-to-solvent ratio of about 1 : 10-1 : 15 (mg per mu L). The combination of tissuelyser homogenization with ultrasonication was the choice of cell-breaking method with three repeated extractions being necessary. For NMR analysis, the optimal extract-to-solvent was around 5-8 mg mL(-1) and completely relaxed spectra were ideal for intrinsically quantitative metabolomic analysis although partially relaxed spectra were employable for comparative metabolomics. This optimized method will offer ensured data quality for high-throughput and reliable plant metabolomics studies.
WOS标题词	Science & Technology ; Physical Sciences
类目[WOS]	Chemistry, Analytical
研究领域[WOS]	Chemistry
关键词[WOS]	MULTIVARIATE DATA-ANALYSIS ; SPINNING H-1-NMR SPECTROSCOPY ; NUCLEAR-MAGNETIC-RESONANCE ; METABONOMIC ANALYSIS ; LIQUID-CHROMATOGRAPHY ; HIGH-TEMPERATURE ; RESPONSES ; IDENTIFICATION ; TISSUES ; SYSTEMS
收录类别	SCI
语种	英语
WOS记录号	WOS:000332422400027
公开日期	2015-07-14
内容类型	期刊论文
源URL	[http://ir.wipm.ac.cn/handle/112942/1443]
专题	武汉物理与数学研究所_磁共振应用研究部
作者单位	1.Univ Chinese Acad Sci, Key Lab Magnet Resonance Biol Syst, State Key Lab Magnet Resonance & Atom & Mol Phys, Wuhan Ctr Magnet Resonance,Wuhan Inst Phys & Math, Wuhan 430071, Peoples R China 2.Fudan Univ, Sch Life Sci, State Key Lab Genet Engn Biospectroscopy & Metab, Shanghai 200433, Peoples R China
推荐引用方式 GB/T 7714	Wu, Xiangyu,Li, Ning,Li, Hongde,et al. An optimized method for NMR-based plant seed metabolomic analysis with maximized polar metabolite extraction efficiency, signal-to-noise ratio, and chemical shift consistency[J]. ANALYST,2014,139(7):1769-1778.
APA	Wu, Xiangyu,Li, Ning,Li, Hongde,&Tang, Huiru.(2014).An optimized method for NMR-based plant seed metabolomic analysis with maximized polar metabolite extraction efficiency, signal-to-noise ratio, and chemical shift consistency.ANALYST,139(7),1769-1778.
MLA	Wu, Xiangyu,et al."An optimized method for NMR-based plant seed metabolomic analysis with maximized polar metabolite extraction efficiency, signal-to-noise ratio, and chemical shift consistency".ANALYST 139.7(2014):1769-1778.