DOI: 10.1002/pca.3271 ISSN: 0958-0344

High‐speed countercurrent chromatography with offline detection by electrospray mass spectrometry and nuclear magnetic resonance detection as a tool to resolve complex mixtures: A practical approach using Coffea arabica leaf extract

Aline Camargo Jesus de Souza Wuillda, Fernanda das Neves Costa, Rafael Garrett, Mauro dos Santos de Carvalho, Ricardo Moreira Borges
  • Complementary and alternative medicine
  • Drug Discovery
  • Plant Science
  • Molecular Medicine
  • General Medicine
  • Biochemistry
  • Food Science
  • Analytical Chemistry

Abstract

Introduction

Many secondary metabolites isolated from plants have been described in the literature owing to their important biological properties and possible pharmacological applications. However, the identification of compounds present in complex plant extracts has remained a great scientific challenge, is often laborious, and requires a long research time with high financial cost.

Objectives

The aim of this study was to develop a method that allows the identification of secondary metabolites in plant extracts with a high degree of confidence in a short period of time.

Material and methods

In this study, an ethanolic extract of Coffea arabica leaves was used to validate the proposed method. Countercurrent chromatography was chosen as the initial step for extraction fractionation using gradient elution. Resulting fractions presented a variation of compounds concentrations, allowing for statistical total correlation spectroscopy (STOCSY) calculations between liquid chromatography coupled with high‐resolution tandem mass spectrometry (LC‐HRMS/MS) and NMR across fractions.

Results

The proposed method allowed the identification of 57 compounds. Of the annotated compounds, 20 were previously described in the literature for the species and 37 were reported for the first time. Among the inedited compounds, we identified flavonoids, alkaloids, phenolic acids, coumarins, and terpenes.

Conclusion

The proposed method presents itself as a valid alternative for the study of complex extracts in an effective, fast, and reliable way that can be reproduced in the study of other extracts.

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