Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/65990
標題: 季節對台灣產細本、光葉及基隆葡萄莖部萃取物成分與抗氧化活性之影響
Seasonal Effects on Ingredients and Antioxidant Activities of Stem Extracts from Vitis thunbergii, Vitis flexuosa, and Vitis kelungensis in Taiwan
作者: 鄭凱中
Cheng, Kai-Chung
關鍵字: Wild-grape;野生葡萄;Season;Antioxidant;Index compound;Resveratrol derivatives;季節;抗氧化;指標成分;白藜蘆醇衍生物
出版社: 森林學系所
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摘要: 
台灣常見的野生葡萄主要有台灣原生種細本葡萄(Vitis thunbergii)、光葉葡萄(Vitis flexuosa)及台灣特有種基隆葡萄(Vitis kelungensis)三種。台灣野葡萄為民間珍貴的保健作物,其宣稱具有祛風與保肝等功效,其中,細本葡萄與光葉葡萄莖部抽出物中已證實含有白藜蘆醇類化合物。然而,對於台灣野葡萄所潛藏具生物活性之抽出成分,截至目前為止仍缺乏相關性研究。因此,本研究針對不同採集季節之細本、光葉及基隆葡萄莖部甲醇粗萃物,利用DPPH自由基清除試驗、超氧自由基清除試驗、亞鐵離子螯合試驗、還原力試驗及總酚含量測定等試驗,評估其抗氧化活性差異。此外,並利用管柱層析與高效能液相層析等技術,分離及純化抽出成分中之指標成分,同時亦運用質譜儀及核磁共振分析儀等儀器,分析與鑑定其成分之確切結構。
試驗結果顯示,不同採集季節之細本、光葉及基隆葡萄莖部甲醇粗萃物之抗氧化活性,以冬季採集之基隆葡萄具有最佳抗氧化效果,其莖部甲醇粗萃物對DPPH自由基清除活性及超氧自由基清除活性均與常見抗氧化劑-兒茶素相當。而冬季採集之基隆葡萄莖部甲醇粗萃物及其各可溶部之活性表現,則以正丁醇可溶部具有最佳清除DPPH自由基與超氧自由基之效果,其IC50值分別為2.9 μg/mL與7.7 μg/mL。同樣的,在還原力與總酚含量方面,亦以正丁醇可溶部具有最佳表現,其值分別為600.1 mg CE/g與330.6 mg GAE/g。
此外,基隆葡萄莖部指標成分分析方面,經分離、純化及鑑定後,共獲得10種主要化合物,其中包括3種植物固醇(β-Sitostenone、β-Sitosterol及Stigmasterol)、4種三萜類化合物(Lupeol、β-Amyrin、Betulinic acid及Betulin)以及3種白藜蘆醇衍生物((-)-ε-Viniferin、2-(4-Hydroxyphenyl)-2,3-dihydrobenzo[b]furan-3,4,6-triol及 (+)-Balanocarpol);其中,2-(4-Hydroxyphenyl)-2,3-dihydrobenzo[b]furan-3,4,6 -triol為首次被分離鑑定之白藜蘆醇類化合物。此外,(-)-ε-Viniferin於不同採集季節之細本、光葉及基隆葡萄莖部,其每克氣乾重之莖部含量分別為2.20-0.41 mg/g、1.47-0.41 mg/g以及1.54-0.66 mg/g。三種葡萄之中,細本葡萄於春季採集時(-)-ε-Viniferin含量最高,而夏季採集時含量則最低。此外,光葉葡萄則是秋季採集時含量最高,而冬季採集時含量最低。至於基隆葡萄則是於冬季採集時含量最高,而秋季含量最低。另外,2-(4-Hydroxyphenyl)- 2,3-dihydrobenzo[b]furan-3,4,6-triol則僅存在於細本葡萄與基隆葡萄莖部;其中,細本葡萄亦於春季採集時含量最高(1.96 mg/g),而基隆葡萄則是於春季採集時含量最高(0.89 mg/g)。至於(+)-Balanocarpol則僅存在於光葉葡萄與基隆葡萄莖部,且二者均於夏季採集時含量最高,其含量分別為0.14 mg/g與0.11 mg/g。

Vitis thunbergii, Vitis flexuosa, and Vitis kelungensis are three common wild-grapes in Taiwan. They have been used as a health protection crop to prevent some diseases, such as flu and hepatitis, etc. It is well-known that V. thunbergii and V. flexuosa stems contain various resveratrols. However, the potential bioactivities of those wild-grapes have not been studied to data. Thus, in this study, the antioxidant activities of methanolic extracts from V. thunbergii, V. kelungensis, and V. flexuosa stems in different seasons are evaluated by various in vitro antioxidant models, including DPPH radical scavenging assay, superoxide radical scavenging assay, ferrous ion chelating ability assay, reducing power assay, and total phenolic content assay. In addition, column chromatography (CC) and high performance liquid chromatography (HPLC) are employed to separate and purify maker ingredients of extracts from Vitis kelungensis, and identified by the analyses of mass spectroscopy and nuclear magnetic resonance spectroscopy (NMR), etc.
Results revealed that, among all tested samples, V. kelungensis harvested in winter exhibited the strongest antioxidant activities in DPPH radical scavenging activity, superoxide radical scavenging activity, and ferrous ion chelating ability, as well as its antioxidant activities were similar to (+)-catechin, a well-known antioxidant. Furthermore, among all derived soluble fractions from the methanolic extracts of V. kelungensis stem, the BuOH fraction exhibited the strongest DPPH radical and superoxid radical scavenging activities with IC50 values of 2.9 μg/mL and 7.7 μg/mL, respectively. Additionally, the BuOH fraction also exhibited the highest reducing power (600.1 mg CE/g) and total phenolic content (330.6 mg GAE/g).
Following by column chromatography and high performance liquid chromatography, 10 major constituents, including 3 phytosteroids (β-sitostenone, β-sitosterol and stigmasterol), 4 triterpenoids (lupeol, β-amyrin, betulinic acid and betulin), and 3 resveratrol derivatives ((-)-ε-viniferin, 2-(4-hydroxyphenyl)-2,3- dihydrobenzo[b]furan-3,4,6-triol and (+)-balanocarpol), were isolated and purified from the EtOAc fraction of V. kelungensis stem. Among them, 2-(4-hydroxyphenyl)- 2,3-dihydrobenzo[b]furan-3,4,6-triol was a new compound. On the other hand, the contents of (-)-ε-viniferin were ranged from 0.63 to 2.20 mg, 0.44 to 1.47 mg, and 0.66 to 1.54 mg per gram of aur-dried stems of V. thunbergii, V. flexuosa, and V. kelungensis , respectively. Accordinglly, in spring, V. thunbergii had the highest amount of (-)-ε-viniferin content, whereas, the lowest content of that was obtained in summer, while, the V. flexuosa stem, the highest (-)-ε-viniferin content was obtained in autumn, whereas the lowest content was found in winter. As for V. kelungensis stem, the higest (-)-ε-viniferin content was obtained in winter and spring, while the lowest content was found in autumn. Furthermore, a new compound, 2-(4-hydroxyphenyl)-2,3-dihydro- benzo[b]furan-3,4,6-triol, was only presented in V. thunbergii and V. kelungensis stems. Both species the highest amount of this new compound in spring with values of 1.96 mg/g and 0.89 mg/g, respectively. Similarly, (+)-balanocarpol was only presented in V. kelungensis and V. flexuosa stems. Among all seasons, these two species showed the highest amount of (+)-balanocarpol content in summer with value of 0.17 mg/g and 0.11 mg/g, respectively.
URI: http://hdl.handle.net/11455/65990
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