The Bioassay-Guided Fractionation and Identification of Potent Acetylcholinesterase Inhibitors from Narcissus c.v. 'Hawera' Using Optimized Vacuum Liquid Chromatography, High Resolution Mass Spectrometry and Bioautography.

Tomasz Mroczek,Krzysztof Kamil Wojtanowski,Jarosław Widelski,Aleksandra Dymek

doi:10.3390/metabo10100395

Tomasz Mroczek, Krzysztof Kamil Wojtanowski + Show 2 more

Open Access

https://doi.org/10.3390/metabo10100395

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Journal: Metabolites	Publication Date: Oct 4, 2020
Citations: 16	License type: CC BY 4.0

Affiliation: Medical University of Lublin

Abstract

Bioassay-guided isolation of bioactive compound is a modern and efficient technique in metabolites screening. It may shorten the total time of the entire process and reduce some costs of it. The aim of this paper was to fractionate and isolate alkaloids by developing an innovative vacuum liquid chromatography method for a species of Narcissus c.v. ‘Hawera’ rarely investigated so far and establishing the inhibitory activity of acetylcholinesterase (AChE). The studies consisted of the extraction of plant material by modern pressurized liquid extraction (PLE), followed by the isolation of alkaloidal fractions. For this purpose, the pioneering gradient vacuum liquid chromatography (gVLC) technique was employed by using two sorbents in various proportions packed in polypropylene cartridges for the first time. This step was performed in order to pre-clean the samples but also to establish the best combination of sorbents which permits obtaining potentially strong AChE inhibitors. The collected fractions were examined by HPLC/ESI-QTOF-MS in order to compare which combination of sorbents would allow us to obtain the highest concentration of alkaloids. The combination of these techniques confirmed the presence of the alkaloids and enabled the development of a modern method for the fractionation and isolation of the compounds with strong anti-AChE activity.

Highlights

Nature provides a huge diversity of compounds, which can be used for example as anticancer agents [1] or as a source of antioxidant polyphenolic ingredients of functional food [2]
The results obtained in research mentioned above clearly shows, that Narcissus ‘Hawera’ among the others plants from genus Narcissus and, generally, Amaryllidaceae family is very rich source of bioactive alkaloids
The gradient vacuum liquid chromatography (gVLC) method proved to be an efficient technique for purifying sample, concentration, fractionation and preparing for subsequent analysis

Summary

Introduction

Nature provides a huge diversity of compounds, which can be used for example as anticancer agents (curcuminoids from Curcuma longa) [1] or as a source of antioxidant polyphenolic ingredients of functional food (e.g., chokeberry-Aronia melanocarpa) [2]. Alkaloids are the group of substances occurring often in nature both in plant and animal kingdoms. They play very important role mainly due to their strong antioxidant, anxiolytic and antidepressant properties [7]. They have been used in the treatment of neurodegenerative diseases [7].

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