Quantitative Analyses of Galanthamine of In Vitro and In Vivo Plant Bulbs of Lycoris aurea Herb. by Capillary Column Gas Chromatography

• Main Authors: Taing-Yoe Dai, 戴天佑
• Other Authors: Li-Chun Huang
• Format: Others
• Language: zh-TW
• Published: 2001
• Online Access: http://ndltd.ncl.edu.tw/handle/05154538796427985551

碩士 === 台北醫學院 === 生藥學研究所 === 89 === Biologically Galanthamine is an alkaloid and belongs to a secondary metabolite, which is wildly distributed in Amaryllidaceae family. It exerts cholinesterase inhibitory effect and has been used in clinical trials against Alizheimer's disease pharmaceutically. In this investigation, capillary gas chromatography-NPD spectrometry was employed to detect galanthamine in Lycoris aurea plants in vitro and in vivo. This system was operated under the following conditions: an Ultra-1 Capillary Column (15m ×0.25mm i.d.×0.2mm film thickness；Hewlett Packard) was coupled to the NPD (Nitrogen Phosphorus Detector). The carrier gas was Nitrogen at a constant flow rate of 1.2 ml/min. The temperatures were 260℃ for the injector, 260℃ for the detector, and 200℃ for initial column , The column temperature was linear increased with 4℃/min to reach to 250℃，and held for 3 minutes. The sensitivity can be detected as low as the concentration of 1.5625 g/ml in the standard sample. The sample preparation was first to dry the bulbs or entire plants in a oven at 80℃，then stored in the desecrator in the powder form. A mount of 500mg powder was first extracted in 6ml of 0.05N HCl solution at 40℃ for 1hour, then added 1ml 0.3N NaOH to neutralize, and partition with 3ml chloroform. Dry the extract with the aid of Nitrogen gas and dissolved in 100μl methanol. Determination the content of galanthamine in Lycoris aurea plants established in vitro and field growing in 5 areas in Taiwan from north to south., i.e., 宜蘭、淡水、桃園、后里、台東. Capillary GC analyses indicated the production of galanthamine in bulbs collected in April and August separately was not correlated in terms of bulb sizes and galanthamine yield. The galanthamine yields of tissue culture plants and bulbs were 132.99μg及70.48μg per gram dry weight respectively annually. The field growing plants in five different areas synthesized the galanthamine 9.69 to 138.85μg per gram dry weight of 3-4 years old bulbs. The superiority of galanthamine content from tissue culture plants probably was due to the shoot apical meristem established cultures or clones possessed the pathogen —free character.