Highly efficient,selective and sensitive molecular screening of acetylcholinesterase inhibitors of natural origin by solid-phase extraction-liquid chromatography/electrospray ionisation-octopole-orthogonal acceleration time-of-flight-mass spectrometry and novel thin-layer chromatography-based bioautography |
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Authors: | Tomasz Mroczek |
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Institution: | Department of Pharmacognosy with Medicinal Plant Laboratory Unit, 1 Chod?ki St., 20-093 Lublin, Poland |
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Abstract: | Highly efficient, selective and sensitive molecular screening of natural acetylcholinesterase (AChE) inhibitors was developed and comprised optimized pressurized liquid extraction (PLE) of plant materials followed by highly selective solid-phase extraction (SPE) using Oasis HLB cartridges. Pure alkaloidal fractions were analyzed by a newly developed high-performance liquid-chromatography (HPLC) on a 3 μm Atlantis HILIC silica stationary phase combined with recently introduced electrospray ionisation (ESI) octopole–orthogonal acceleration time-of-flight (oa TOF)-mass spectrometry (MS) with high mass accuracy (about 2 ppm) and high sensitivity (absolute limit of detection (LOD) for galanthamine was about 43 fg at signal-to-noise 13:1). Moreover, a newly developed and validated TLC-bioautography permit galanthamine sensitivities at pg levels. In this way, more potent than galanthamine AChE inhibitor namely 1,2-dihydrogalanthamine in Narcissus jonquilla ‘Pipit’ extract could be found (with IC50 value 0.19 μM lower of about 42% than that of galanthamine). |
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Keywords: | High-performance liquid chromatography Time-of-flight mass spectrometry Bioauthography Acetylcholinesterase inhibitors Amaryllidaceae alkaloids |
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