Robust standard procedures for plant metabolomics with NMR-improvements in the extraction and processing procedures

J Schripsema; M Almeida Lemos; M de Fatima Dianin Vianna; D Soares Vianna; D Saraiva Dagnino

doi:10.1055/s-0032-1321170

Planta Medica, Table of Contents

Robust standard procedures for plant metabolomics with NMR-improvements in the extraction and processing procedures

J Schripsema ¹, M Almeida Lemos ¹, M de Fatima Dianin Vianna ¹, D Soares Vianna ², D Saraiva Dagnino ³

¹Grupo Metabolômica,Laboratório de Ciências Quimicas, Universidade Estadual do Norte Fluminense, 28013–602 Campos dos Goytacazes, RJ,Brazil
²Universidade Federal Fluminense, Rio das Ostras, RJ, Brazil
³Laboratório de Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ, Brazil

Abstract

In metabolomics comprehensive metabolic fingerprints are obtained and subsequently analyzed to find the metabolites which differentiate groups of samples.

A new extraction method with a two-phase system of water and chloroform was evaluated and validated, by varying different variables in the extraction protocol (e.g. the extraction time and cycles of ultrasonic vibration). The effect of these variables was tested on the yield of the secondary metabolite eupatorin, a major secondary metabolite in plant material of Baccharis trimera.

For preprocessing the NMR data before submission to multivariate data processing, a specific software has been developed for reference deconvolution. The signal from an internal standard is used for the determination of the experimental peak shape. Subsequently all signals in the spectrum are corrected for this peak shape, reducing the peaks to single lines. A great reduction in data points is obtained and furthermore small signals are preserved, even when they are not resolved from major peaks.