Partial least squares model and design of experiments toward the analysis of the metabolome of Jatropha gossypifolia leaves: Extraction and chromatographic fingerprint optimization

Author(s)

Pilon, Alan Cesar ^[1] ; Neto, Fausto Carnevale ^[1] ; Freire, Rafael Teixeira ^[1] ; Cardoso, Patricia ^[1] ; Carneiro, Renato Lajarim ^[2] ; Bolzani, Vanderlan Da Silva ^[1] ; Castro-Gamboa, Ian ^[1]

Affiliation

^[1] UNESP, Inst Chem, Dept Organ Chem, Biosynth & Ecophysiol Nat Prod NuBBE, Nucleus Bioa, Rua Prof Francisco Degni 55 CP 355, BR-14800900 Araraquara, SP - Brazil

^[2] Sao Carlos Fed Univ UFSCar, CCET, Dept Chem, Rodovia Washington Luiz, Sao Carlos, SP - Brazil

• Document type: Journal article
• Source: JOURNAL OF SEPARATION SCIENCE; v. 39, n. 6, p. 1023-1030, MAR 2016.
• Web of Science Citations: 5

Abstract

A major challenge in metabolomic studies is how to extract and analyze an entire metabolome. So far, no single method was able to clearly complete this task in an efficient and reproducible way. In this work we proposed a sequential strategy for the extraction and chromatographic separation of metabolites from leaves Jatropha gossypifolia using a design of experiments and partial least square model. The effect of 14 different solvents on extraction process was evaluated and an optimized separation condition on liquid chromatography was estimated considering mobile phase composition and analysis time. The initial conditions of extraction using methanol and separation in 30 min between 5 and 100% water/methanol (1:1 v/v) with 0.1% of acetic acid, 20 L sample volume, 3.0 mL min(-1) flow rate and 25 degrees C column temperature led to 107 chromatographic peaks. After the optimization strategy using i-propanol/chloroform (1:1 v/v) for extraction, linear gradient elution of 60 min between 5 and 100% water/(acetonitrile/methanol 68:32 v/v with 0.1% of acetic acid), 30 L sample volume, 2.0 mL min(-1) flow rate, and 30 degrees C column temperature, we detected 140 chromatographic peaks, 30.84% more peaks compared to initial method. This is a reliable strategy using a limited number of experiments for metabolomics protocols. (AU)

• FAPESP's process: 13/07600-3 - CIBFar - Center for Innovation in Biodiversity and Drug Discovery
• Grantee: Glaucius Oliva
• Support type: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 10/17935-4 - Development of analytical methods of dereplication by NMR and multivaried analysis of metabolomic profile from Solanaceae species with bioactive potential
• Grantee: Alan Cesar Pilon
• Support type: Scholarships in Brazil - Doctorate