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液態發酵香杉芝菌絲體活性成分Antrodin C在超臨界流體萃取及模擬移動床層析純化研究

Supercritical Fluid Extraction of Antrodin C from Antrodia salmonea Mycelia and Subsequent Isolation by Simulated Moving Bed Chromatography

摘要


香杉芝為臺灣特有真菌,寄生於臺灣特有的香杉樹(Cunninghamia Konishii)枯樹幹中,香杉芝與樟芝同屬多孔菌科(Polyporaceae),薄孔菌屬(Antrodia),兩者外型極為相似。液態發酵的香杉芝菌絲體中發現許多的馬來酸衍生物,其中的潛力活性成分Antrodin C,為具有抗癌幹細胞、護肝、可預防或治療糖尿病相關的心血管疾病、阿茲海默症等活性。本研究目的為開發超臨界流體(SF)萃取及模擬移動床(SMB)層析方法,量產純化香杉芝活性成分Antrodin C。以SF萃取時,採用95%乙醇當作輔溶劑,粗萃率達13.6%,在萃取過程中前0.5 h收集得到的萃取液,可做為高活性成分的商業產品;收集萃取第0.5 h~2 h收集得到的萃出物,則繼續做為下一階段SMB層析純化的中間原料。SMB的組態設計為1-2/2/3,可以有效地將固定相再生,使用管柱尺寸為5 × 30 cm,進料溶液濃度50.00 g/L,在切換時間設定為5.5分鐘時,進料樣品回收率達99%以上,Antrodin C純度達74.39%。研究結果顯示,在超臨界流體萃取後,經SMB純化其萃取物的最大處理量可達550公斤/年,相當適合邁向工業化量產製備,並能以更經濟環保的方式提供活性原料。

並列摘要


Antrodia salmonea (A. salmonea) is a Taiwan-specific fungus that parasitizes the dead trunk of the Taiwan-specific Cunninghamia Konishii tree. Both Antrodia cinnamomea and A. salmonea are fungi of the Polyporaceae family and therefore are morphologically similar. In the liquid A. salmonea mycelium, many maleic acid derivatives were found. Among them, the potential active compound, Antrodin C, has been shown to exhibit anti-cancer stem cells, hepatoprotective, cardiovascular protective, and anti-Alzheimer's disease. Hence, the purpose of this research is to develop supercritical fluid (SF) extraction and simulated moving bed (SMB) chromatography methods to mass-produce and purify the active ingredient Antrodin C from A. salmonea. When SF is used for extraction, 95% ethanol is used as the solvent, the crude extraction rate reaches 13.6%.Within the first 0.5 h of extraction, the A. salmonea extract can be sold as a commercial product with high active ingredients (Antrodin C). Between 0.5 h to 2 h of extraction, the A. salmonea extract could be used as the intermediate raw material for subsequent chromatographic purification. The configuration design for SMB is 1-2/2/3, which can effectively regenerate the stationary phase. The column size used was 5 × 30 cm, and the concentration of the feed solution was 50.00 g/L. When the switching time was set to 5.5 minutes, the sample has over a 99% recovery rate, Antrodin C has a purity of 74.39%. Taken together, these results showed that using SF for extraction, the maximum amount of the A. salmonea extract after SMB purification can reach up to 550 kg/year, and the raw materials with active compounds can be produced in a more economically sustainable manner, which can therefore create competitive advantages for industrial production.

參考文獻


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