Electrospray mass-spectrometry guided target isolation of neolignans from Nectandra leucantha (Lauraceae) by high performance- and spiral-coil countercurrent chromatography

Author(s)

Grecco, Simone dos Santos ^{[1, 2]} ; Letsyo, Emmanuel ^[3] ; Tempone, Andre Gustavo ^[4] ; Ghilardi Lago, Joao Henrique ^[1] ; Jerz, Gerold ^[5]

Affiliation

^[1] Fed Univ ABC, Ctr Nat Sci & Humanities, BR-09210580 Santo Andre, SP - Brazil

^[2] Anhanguera Univ Sao Paulo, Biotechnol & Innovat Hlth & Pharm Postgrad Progra, BR-05145200 Sao Paulo, SP - Brazil

^[3] Ho Tech Univ, Dept Food Sci & Technol, POB HP 217, Ho - Ghana

^[4] Adolfo Lutz Inst, Ctr Parasitol & Mycol, BR-01246902 Sao Paulo, SP - Brazil

^[5] Tech Univ Carolo Wilhelmina Braunschweig, Inst Food Chem, Schleinitzstr 20, D-38106 Braunschweig - Germany

• Document type: Journal article
• Source: Journal of Chromatography A; v. 1608, DEC 20 2019.
• Web of Science Citations: 0

Abstract

Nectandra leucantha (Lauraceae) is a tree indigenous to the tropical Atlantic forests of Brazil, one of the most biodiverse flora hotspots worldwide. This plant species contains high concentrations of neolignan and dehydrodieugenol derivatives that express significant in-vitro activities against various parasite strains. These activities are however responsible for severe tropical human infections, such as Leishmaniasis (Leishmania spp.) and Chagas disease (Trypanosoma cruzi), which have been classified by the World Health Organization (WHO) as Neglected Tropical Diseases (NTDs). In order to optimize the isolation process for these target metabolites, n-hexane extract of the leaves was separated by means of semi-preparative high performance countercurrent chromatography (HPCCC) and scale-up spiral-coil countercurrent chromatography (sp-CCC) systems. Several biphasic solvent mixtures were evaluated for their partitioning effects on neolignans, resulting in the selection of an optimized system n-hexane - ethylacetate - methanol - water (7:3:7:3, v/v/v/v). The chromatographic experiments on the HPCCC and sp-CCC were run in the head-to-tail mode with 500 mg and 16 g injections, respectively. For specific and multiple metabolite detection, the recovered CCC-fractions were off-line injected, in the sequence of recovery, to an electrospray mass-spectrometry (ESI-MS/MS) device. A projection of the single ion traces of the target compounds, in the positive ionization mode at a scan range of m/z 100-1500, located chromatographic areas where the co-elution effects occurred and pure target metabolites were present. Five major target neolignans were specifically detected, which enabled the accurate pooling of CCC-fractions for an optimum recovery of the metabolites. The direct comparison of the performance characteristics of the two CCC-devices, with very different mechanical designs was achieved by the conversion of the time axis into a partition ratio (K-D) separation scale. As a result, the compound specific K-D-elution values of the target neolignan were determined in high precision, while the comparison of the calculated separation factor (alpha) and resolution factor (R-s) values revealed a superior separation performance for the HPCCC system. Also, the reproducibility of detected metabolites in the two CCC experiments was confirmed by small variations (Delta K-D +/- 0.1). Neolignan target compounds with anti-parasite activities were successfully isolated in the 100 mg to 4 g range in a single lab-scale countercurrent chromatographic process step. (C) 2019 Published by Elsevier B.V. (AU)

• FAPESP's process: 18/07885-1 - Biomolecules from plant species of remnant areas of the Atlantic Forest and Cerrado to treat neglected tropical diseases - chemical and pharmacological aspects
• Grantee: João Henrique Ghilardi Lago
• Support Opportunities: BIOTA-FAPESP Program - Regular Research Grants
• FAPESP's process: 18/09083-0 - The 10th International Conference on Countercurrent Chromatography
• Grantee: Simone dos Santos Grecco
• Support Opportunities: Research Grants - Meeting - Abroad