舞菇T2品系乙酸乙酯萃取層之生物活性成份分析

Bioactive Component Analysis of Ethyl Acetate Extract from Grifola frondosa

方苡藻(Yi-Zao Fang)

指導教授 ： 張竣維

亞洲大學/健康學院/生物科技學系碩士班/碩士(2011年)

摘要

本研究主要分析環控技術所栽培之舞菇(Grifola frondosa)所含之生物活性成份。以正己烷、乙酸乙酯與50%酒精分別進行超音波萃取乾燥粉末樣品，其萃取率分別為1.60 ± 0.36%、3.10 ± 0.17%與22.90 ± 0.40%。測試不同粗萃物之DPPH自由基清除能力、還原力與亞鐵離子螯合能力，結果顯示粗萃物於三種抗氧化測試中皆有良好效果。利用高效能液相層析質譜儀(High-performance Liquid Chromatography-Mass Spectrophotometer)檢測各萃取層內之成份，於乙酸乙酯萃取層與正己烷萃取層共測得7個分離峰，取其分離峰相對純化物進行三種抗氧化能力測試，結果顯示S4與S5具有清除DPPH自由基的能力(EC90各為0.96 ± 0.02mg/ml及0.04 ± 0.02mg/ml)，以S5清除自由基的能力最好；還原力測定結果顯示S4、S5均具有還原力，S4之還原力約為BHT的36.08 ± 1.36%，S5之還原力則高於BHT(>100%)；亞鐵離子螯合能力的測試結果，則發現五種成份並不具有亞鐵離子螯合能力；經由酪胺酸酶的測試則推斷其抑制型態為不競爭性抑制。

關鍵字

舞菇 ； 超音波萃取 ； DPPH自由基清除能力 ； 還原力測定 ； 亞鐵離子螯合能力 ； 高效能液相層析質譜儀 ； 酪胺酸酶

並列摘要

The aim of this study was to analyze bioactivities of components extracted from Grifola frondosa that was grown in environmental controlled cultivation conditions. Ultrasonic extraction was used to extract ground powder from dry fruit body of Grifola frondosa by n-hexane, ethyl acetate and 50% ethanol. The extraction efficiencies of various organic solvents were 1.60 ± 0.36%, 3.10 ± 0.17% and 22.90 ± 0.40% for n-hexane, ethyl acetate and 50% ethanol respectively. DPPH radical scavenging activity, reducing power and ferrous ion chelating activity assays were used to analyze the bioactivity of various organic solvent extracts. Results showed that all extracts containing antioxidant activity. The components of various organic solvent extracts were detected by high-performance liquid chromatography-mass spectrophotometry (LC-MS/MS). Totally, seven peaks in ethyl acetate extract and n-hexane extract including S1-S7 were observed. Three antioxidant activity assays were used to determine the purified isolates. Results showed that S4 and S5 both have 90% DPPH radical scavenging activities with EC90=0.96 ± 0.02mg/ml for S4 and EC90=0.04 ± 0.02mg/ml for S5 respectively. Results of reducing power assay showed S4 had 36.08 ± 1.36% reducing power activitycompared with BHT, and the reducing power activity of S5 (>100%) which was higher than BHT. Interestingly, the crude ethyl acetate and n-hexane extracts of Grifola frondosa showed efficacy in ferrous ion chelating activities, but S1-S5 showed no efficacy in ferrous ion chelating activity. Finally, Lineweaver-Burk plots showed that S4 and S5 were both non-competitive inhibitors of L-Dopa for tyrosinase mediated oxidative activity.

並列關鍵字

Grifola frondosa ； ultrasonic extraction ； DPPH radical scavenging activity ； reducing power assay ； ferrous ion chelating activity ； high-performance liquid chromatography-mass spectrophotometry ； non-competitive inhibitors ； L-Dopa ； tyrosinase mediated oxidative activity

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