Metabolomic analysis of Ocotea odorifera Cell Cultures: A model protocol for acquiring metabolite data

Abstract

Metabolomics constitutes a quantitative and qualitative survey of the whole metabolites of an organism as well as a tissue, reflecting the genome and proteome of a sample as analyzed. Advanced analytical spectroscopic and chromatographic techniques are used along with uni-or multivariate statistical data analysis, rapidly identifying up-or down-regulated metabolites in complex matrices. In this chapter, protocols for the analysis of target compounds (protocol I) and metabolomics (protocol II) of Ocotea odorifera cell cultures are described. In the first case, the target compound safrole, an aromatic ether used as a flavoring agent and also in the manufacture of insecticides, is analyzed in the organosolvent fraction of stable prototrophic cell lines of O. odorifera by gas chromatography-mass spectrometry. For metabolomics studies the protocol is designed to detect and quantify metabolites in the aqueous extract of O. odorifera cell lines by using high-resolution 1D-and 2D-nuclear magnetic resonance spectroscopy, followed by chemometric analysis of the 1H NMR spectra dataset. Protocol I has been successfully used, for example, in screening studies of cell lines able of producing safrole. Protocol II is suitable to detect the chemical features of a number of metabolite compounds in aqueous extracts of O. odorifera cell lines cultured under certain conditions, leading to new insights into metabolomics of that species.