Extraction and isolation of acetylcholinesterase inhibitors from Citrus limon peel using an in vitro method

Abstract

A simple and efficient ultrafiltration-liquid chromatography-mass spectrometrybased method was developed for the rapid screening and identification of ligands from Citrus limon peel, which are suitable acetylcholinesterase inhibitors. Subsequently, the anti-Alzheimer's activity of these compounds was assessed using a PC12 cell model. Six major compounds, viz. neoeriocitrin, isonaringin, naringin, hesperidin, neohesperidin, and limonin, were identified as potent acetylcholinesterase inhibitors.A continuous and efficient online method, which involved the use of a microwaveassisted extraction device, solvent concentration tank, and centrifugal partition chromatography column, was developed for the scale-up of these compounds, and the obtained compounds presented high purity. Next, their bioactivity was evaluated using a PC12 cell model. This novel approach, which was based on ultrafiltration-liquid chromatography-mass spectrometry, microwave-assisted extraction online coupled with solvent concentration tank, and centrifugal partition chromatography along with in vitro evaluation, could represent a powerful tool for the screening and extraction of acetylcholinesterase inhibitors from complex matrices, and could be a useful platform for the large-scale production of bioactive and nutraceutical ingredients.

Single‐step screening and isolation of potential lipoxidase inhibitors from Trifolium repens by stepwise flow rate high‐speed countercurrent chromatography and semipreparative high‐performance liquid chromatography target‐guided by ultrafiltration‐LC‐MS

“…The choice of the solvent system is critical in stepwise flow rate HSCCC. According to a previous study, the most suitable K value range for HSCCC is 0.5–2.0 [24].…”

Food‐derived Acetylcholinesterase Inhibitors as Potential Agents against Alzheimer’s Disease

“…In a previous work, the Hawthorn Fruit (HF) peel was extracted with 95% ethanol (PHF) followed by further separation on a macroporous resin column to obtain a 20% ethanol eluate (EPHF), both of which were tested as potential AChE inhibitors [40]. [41]. In PC12 cells that were treated with the Amyloidbeta (Ab ) peptide, addition of each identified phenolic compound led to concentration-dependent reduction in cell death.…”

Affinity selection–mass spectrometry for the discovery of pharmacologically active compounds from combinatorial libraries and natural products

“…Anti‐inflammation targets have included cyclooxygenase‐1 23 and cyclooxygenase‐2, 24,25 secretory phospholipase A2, 26 and 5‐lipoxygenase 27 . Parkinson's disease and Alzheimer's disease targets have included tyrosinase 28 and acetylcholinesterase 29 . Antimicrobial, antimalarial, and antiviral targets have included eukaryotic dihydrofolate reductase, 30 Plasmodium falciparum thioredoxin and glutathione reductases, 31 neuraminidase, 32 and Ebola and Marburg virus nucleoproteins 33 .…”

“…27 Parkinson's disease and Alzheimer's disease targets have included tyrosinase 28 and acetylcholinesterase. 29 Antimicrobial, antimalarial, and antiviral targets have included eukaryotic dihydrofolate reductase, 30 Plasmodium falciparum thioredoxin and glutathione reductases, 31 neuraminidase, 32 and Ebola and Marburg virus nucleoproteins. 33 Anticancer targets have included retinoid X receptor (RXR)-α, 34 quinone reductase-2, 22 and urokinasetype plasminogen activator.…”