Sample Preparation Issues in NMR‐based Plant Metabolomics: Optimisation for Vitis Wood Samples1. Issue 4 (4th February 2014)

• Record Type: Journal Article
• Authors: Halabalaki, Maria; Bertrand, Samuel; Stefanou, Anna; Gindro, Katia; Kostidis, Sarantos; Mikros, Emmanuel; Skaltsounis, Leandros A.; Wolfender, Jean‐Luc; Choi, Young Hae; Verpoorte, Robert
• Is Part Of: Phytochemical analysis. Volume 25:Issue 4(2014:Jul.)
• Page Start: 350
• Page End: 356
• Publication Date: 2014-02-04
• Subjects: Plants -- Analysis -- Periodicals; Plants -- chemistry -- Periodicals; 572.2
• DOI: 10.1002/pca.2497
• Languages: English
• ISSNs: 0958-0344

Abstract

<abstract abstract-type="main"> <title>ABSTRACT</title> <sec id="pca2497-sec-0001" sec-type="section"> <title>Introduction</title> <p>Nuclear magnetic resonance (NMR) is one of the most commonly used analytical techniques in plant metabolomics. Although this technique is very reproducible and simple to implement, sample preparation procedures have a great impact on the quality of the metabolomics data.</p> </sec> <sec id="pca2497-sec-0002" sec-type="section"> <title>Objective</title> <p>Investigation of different sample preparation methods and establishment of an optimised protocol for untargeted NMR‐based metabolomics of <italic>Vitis vinifera</italic> L. wood samples.</p> </sec> <sec id="pca2497-sec-0003" sec-type="section"> <title>Methods</title> <p>Wood samples from two different cultivars of <italic>V. vinifera</italic> with well‐defined phenotypes (Gamaret and 2091) were selected as reference materials. Different extraction solvents (successively, dichloromethane, methanol and water, as well as ethyl acetate and 7:3 methanol‐water (v/v)) and deuterated solvents (methanol‐<italic>d</italic>4, 7:3 chloroform‐<italic>d</italic>‐methanol‐<italic>d</italic>4 (v/v), dimethylsulphoxide‐<italic>d</italic>6 and 9:1 dimethylsulphoxide‐<italic>d</italic>6‐water‐<italic>d</italic>2 (v/v)) were evaluated for NMR acquisition, and the spectral quality was compared. The optimal extract concentration, chemical shift stability and peak area repeatability were also investigated.</p> </sec> <sec id="pca2497-sec-0004" sec-type="section"> <title>Results</title> <p>Ethyl acetate was found to be the most satisfactory solvent for the extraction of all representative chemical classes of secondary metabolites in <italic>V. vinifera</italic> wood. The optimal concentration of dried extract was 10 mg/mL and 7:3 chloroform‐<italic>d</italic>‐methanol‐<italic>d</italic>4 (v/v) was the most suitable solvent system for NMR analysis. Multivariate data analysis was used to estimate the biological variation and clustering between different cultivars.</p> </sec> <sec id="pca2497-sec-0005" sec-type="section"> <title>Conclusion</title> <p>Close attention should be paid to all required procedures before NMR analysis, especially to the selection of an extraction solvent and a deuterated solvent system to perform an extensive metabolomic survey of the specific matrix. Copyright © 2014 John Wiley & Sons, Ltd.</p> </sec> </abstract>