Estudo de duas espécies de Eucalipto usando estratégias metabolômicas untargeted e targeted

Author(s)

João Benhur Mokochinski

• Document type: Doctoral Thesis
• Press: Campinas, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia
• Defense date: 2017-06-02

Examining board members

Alexandra Christine Helena Frankland Sawaya; Alessandra Sussulini; Paulo César Pires Rosa; Patricia de Oliveira Carvalho; Marcia Ortiz Mayo Marques

• Advisor: Alexandra Christine Helena Frankland Sawaya

Abstract

Metabolomics analysis aims to identify and quantify the metabolites produced by a given organism, proving to be a useful tool in elucidating the end-products of a biological system. Among the various techniques currently employed, ultra-high-performance liquid chromatography coupled to mass spectrometry (UHPLC-MS) has been shown to be a robust technique for the analysis of metabolites, since it allows a high degree of reproducibility, low solvent consumption and short analysis time. When coupled to a high-resolution mass spectrometer, the field of applications is considerably expanded, allowing its use in different analytical segments. Metabolomics can be divided basically into two branches: untargeted ¿ intended to search for the largest number of compounds in a sample, without specific targets; or targeted ¿ when the identity of a specific metabolite (or group) is known and it is monitored in the sample. Directed by these concepts, herein we propose the study of the metabolome of an important commercial plant ¿ eucalyptus ¿ divided in two parts. The first part of this thesis presents the results obtained through untargeted analyses, carried out during a research internship period at the University of Wageningen ¿ The Netherlands, allowing the identification of possible biomarkers related to the studied species. The second part addresses targeted analyses, which include the development and validation of analytical methods for quantification of primary and secondary metabolites. In this context, different methods were created to determine the S/G (syringyl/guaiacyl) ratio in lignin, analysis of abscisic acid and its derivatives, analysis of amino acids, sugars, organic acids and phenolic compounds. In addition, factorial design was applied to optimize the extraction conditions. All the methods developed were suitable to quantify metabolites in stems and leaves of eucalyptus. We emphasize that the methods developed herein are not restricted to the eucalyptus and may be employed in other plant matrices (AU)

• FAPESP's process: 13/21306-0 - DEVELOPMENT OF ANALYTICAL METHODS BY UPLC-MS FOR THE METABOLOMIC STUDY OF TWO SPECIES OF EUCALYPTUS EXPOSED TO CHANGES IN TEMPERATURE AND CONCENTRATION OF CO2
• Grantee: João Benhur Mokochinski
• Support Opportunities: Scholarships in Brazil - Doctorate