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
Abstract
:1. Introduction
2. Results and Discussion
2.1. Extraction and Optimization of Fractionation by VLC for the Isolation of Alkaloids from Narcissus triandrus L. c.v. ‘Hawera’
2.2. LC-MS Identification of the Isolated Compounds
2.3. TLC-Bioautography for the Detection of Potent AChE Inhibitors
3. Materials and Methods
3.1. Plant Material
3.2. Sample Preparation and Alkaloid Extraction
3.3. Method Optimization of VLC
3.4. LC-MS Identification of the Isolated Compounds
3.5. TLC with Bioautography of Anticholinesterase Activity
4. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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The Number of Experiments | 1-A | 2-B | 3-C |
---|---|---|---|
Type of column | glass | polypropylene cartridge | polypropylene cartridge |
Sorbent filling ratio (Al2O3(150 MeSh): silica gel (60 F254)) | 1:1 Al2O3(25 g): silica gel (57 g) | 1:3 Al2O3(17 g): silica gel (25 g) | 3:1 Al2O3(57 g): silica gel (8 g) |
Fractions obtained | A-19 A-20-22 A-23-24 A-25 A-26 A-27-33 A-34 | B-3 B-4-7 B-8-9 B-10 B-11 B-12 B-13-14 B-15-17 B-18-20 B-21 B-22-25 B-26 B-27-29 B-30 B-31 | C-2 C-3-4 C-5-8 C-9 C-10-12 C-13-14 C-15-16 C-17 |
% of Isolated Compounds to the Total Amount of All Alkaloid Compounds Obtained from Individual Fractions [%] | Fraction Number | ||||||
---|---|---|---|---|---|---|---|
A-19 | A-20-22 | A-23-24 | A-25 | A-26 | A-27-33 | A-34 | |
Sanguinine | 1.2 | ||||||
Lycoramine | 54.3 | 9.9 | 27.5 | 16.4 | 5.6 | ||
Lycorine | 7.2 | 52.9 | 41.2 | 23.6 | 17.8 | 10.0 | 10.0 |
Ungeremine | 9.9 | 16.0 | 16.8 | 18.6 | 22.6 | 14.6 | |
4,N-didehydro-nor-augustamine | 17.7 | 15.5 | 7.2 | 15.9 | 18.1 | 11.9 | |
Tetrahydro-nor-augustamine | 4.7 | 10.7 | 8.8 | 12.5 | 21.4 | 16.6 | |
Mesembrinole | 6.2 | 7.1 | 24.6 | ||||
Galanthamine | 1.4 | 3.3 |
% of Isolated Compounds to the Total Amount of All Alkaloid Compounds Obtained from Individual Fractions [%] | Fraction Number | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
B-10 | B-11 | B-12 | B-13-14 | B-15-17 | B-18-20 | B-21 | B-22-25 | B-26 | B-27-29 | B-30 | B-31 | |
Sanguinine | 3.4 | 9.2 | 9.1 | 5.8 | 7.5 | 1.4 | 2.9 | |||||
Lycoramine | 39.0 | 38.1 | 36.9 | |||||||||
Lycorine | 11.6 | 26.6 | 29.2 | 22.6 | 17.1 | 21.2 | 22.2 | 22.8 | 26.1 | 9.1 | ||
Ungeremine | 7.7 | 4.5 | 5.8 | 9.2 | 7.6 | 7.2 | 6.3 | 11.5 | 9.2 | 24.9 | 26.1 | |
4,N-didehydro-nor-augustamine | 10.8 | 4.7 | 6.6 | 6.0 | 3.6 | 5.9 | 4.0 | 8.1 | ||||
Tetrahydro-nor-augustamine | 5.8 | 3.4 | 4.3 | 4.7 | 0.8 | 0.8 | ||||||
Mesembrinole | 27.4 | 15.6 | 12.1 | 9.0 | 4.6 | 9.7 | ||||||
Haemanthamine | 3.5 | |||||||||||
Lycorine-N-oxide | 0.7 | |||||||||||
Galanthamine-N-oxide | 2.1 | 1.6 | ||||||||||
Galanthamine | 1.9 | 1.7 | ||||||||||
Tazettine | 3.4 | 8.3 | 9.9 | 5.7 |
% of Isolated Compounds to the Total Amount of All Alkaloid Compounds Obtained from Individual Fractions [%] | Fraction Number | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
C-2 | C-3-4 | C-5-8 | C-9 | C-10-12 | C-13-14 | C-15-16 | C-17 | C-18-24 | C-25 | |
Sanguinine | 1.7 | 8.7 | ||||||||
Lycoramine | 7.5 | 26.5 | 53.5 | |||||||
Lycorine | 23.8 | 23.3 | 2.4 | 20.2 | 25.4 | 22.1 | 15.6 | 30.1 | 7.8 | |
Ungeremine | 4.2 | 8.2 | 6.7 | 4.1 | 4.1 | 10.3 | 11.2 | 7.8 | 2.0 | 10.0 |
4,N-didehydro-nor-augustamine | 4.2 | 9.7 | 12.9 | 4.3 | ||||||
Tetrahydro-nor-augustamine | 5.5 | 2.2 | 3.3 | 2.8 | 1.3 | 9.3 | 3.7 | |||
Mesembrinole | 6.7 | 13.4 | 36.2 | 10.3 | 4.3 | |||||
Galanthamine | 5.9 | |||||||||
Lycoramine-N-oxide | 2.1 | 2.4 | ||||||||
Haemanthamine | 12.2 | 6.9 | 3.4 | 2.6 | ||||||
Lycorine-N-oxide | 2.8 |
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Mroczek, T.; Dymek, A.; Widelski, J.; Wojtanowski, K.K. 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. Metabolites 2020, 10, 395. https://doi.org/10.3390/metabo10100395
Mroczek T, Dymek A, Widelski J, Wojtanowski KK. 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. Metabolites. 2020; 10(10):395. https://doi.org/10.3390/metabo10100395
Chicago/Turabian StyleMroczek, Tomasz, Aleksandra Dymek, Jarosław Widelski, and Krzysztof Kamil Wojtanowski. 2020. "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" Metabolites 10, no. 10: 395. https://doi.org/10.3390/metabo10100395