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Title: 大豆屬與千觔拔屬一條根生理活性之比較及其酚類化合物之分離
Comparison on the bioactivity of Glycine tomentella Hayata and Flemingia philipinensis Merr & Rolfe and the separation of the bioactive polyphenols in the Glycine root.
Authors: Ying-Yu Kuo
郭瑩玉
Contributors: NTOU:Department of Food Science
國立台灣海洋大學:食品科學系
Keywords: 環氧合酶;脂氧合酶
glycine;cyclooxygenase;lipoxygenase
Date: 2002
Issue Date: 2011-06-30T07:39:17Z
Abstract: 摘要 金門大豆屬(Glycine tomentella Hayata)一條根為台灣民眾廣泛 使用之本土特有中草藥,業者宣稱金門一條根的效用類似古籍記載 之千觔拔屬(Flemingia philipinensis Merr & Rolfe ),亦具有鎮痛解 熱、舒筋活血等功效,但其藥效及有效成分並無研究報告,故本研 究首先比較金門大豆屬及千觔拔屬一條根的生理活性,並純化大豆 屬一條根95 %乙醇萃取物,鑑定其功能特性及有效成分。 大豆屬一條根之水、50 %乙醇及95 %乙醇粗萃取物,多酚類 含量分別約為千觔拔屬各相關萃取物的4.5、2 及1.5 倍。以產率換 算時,大豆屬之95 %乙醇粗萃取物多酚類含量高於千觔拔屬,其餘 兩種萃取物無顯著差異。掃除自由基DPPH (α, α-diphenyl-β-picrylhydrazyl)、抑制脂質自氧化及抑制脂氧合酶之能 力,皆以大豆屬活性較高。抑制環氧合酶的能力,以千觔拔屬95 % 乙醇粗萃取物高於大豆屬,且為環氧合酶抑制劑indomethacin 之 30,000 倍。 大豆屬一條根95 %乙醇粗萃取物經polyamide、silica gel 管柱 可得十一區分(fractions),對於人血液流變性質的影響,添加大豆 屬純化之十一區分化合物之血液(fluid behavior index, n=0.72∼ 0.78),皆較未添加一條根分離物之血液(n=0.68)更接近牛頓型流 體(n=1),其黏稠指標(fluid consistency, K)為57∼79 cP,較控制 組(98 cP)低,一條根分離物可降低血液黏稠度,而增加血液流動 性。 在抗氧化力方面,由silica gel 管柱所得五區分抑制不飽和脂肪 酸自氧化力高於95%乙醇粗萃取物,IC50 為粗萃取液之15∼70 %; 所有區分掃除自由基之IC50 為95 %乙醇粗萃取物之2∼21 倍。 大豆屬一條根之十一區分對脂氧合酶之抑制力為NDGA (nordihydroguaiaretic acid)的0.36 ~ 1.67 %、95 %乙醇粗萃取物 之2.6 ~ 17.8 倍,其中以第VIII 區分抑制力最高,其再經製備型HPLC 分離、TLC、呈色反應及UV 吸收光譜,可推測為不含glycoside 之 異黃酮,以核磁共振光譜及HPLC 鑑定比對為異黃酮類之daidzein 化合物,其抑制脂氧合酶的能力為第VIII 區分及95 %乙醇粗萃取物 之1.4 及23 倍,故daidzein 為大豆屬一條根有效抑制脂氧合酶的化 合物。
Abstract The I-Tiao-Gung, root of Glycine tomentella Hayata has been used extensively as a herbal medicine in Taiwan. It was claimed to have similar functions in analgesic, anti-pyretic, muscle relaxative and blood-circulation enhancing effects to those of the Flemingia philipinensis Merr & Rolfe. However, no extensive research regarding its bioactivities nor its active compounds was found in literature. The objectives of this study were to compare the bioactivities of the roots of the 2 species, and to separate and identify the effective compounds in the Glycine root. The polyphenols content extracted from Glycine, using water, 50 % ethanol and 95 % ethanol as solvents were 4.5, 2, and 1.5 times of the corresponding extract of the Flemingia, respectively. The yield in dry weight of polyphenols of 95 % ethanolic extract of Glycine was higher than that of Flemingia. The two other solvent extracts showed no significant difference between the 2 types of I-Tiao-Gung. Glycine was higher in free radical-scavenging activity determined with DPPH, reduction in hemoglobin-catalyzed lipid autoxidation and inhibition of the lipoxygenase (LOX)-catalyzed arachidonate oxidation. But the inhibition on the cyclooxygenase-catalyzed arachidonate oxidation was higher for the 95 % ethanolic extract of Flemingia than that of Glycine, and was 30,000 times of the positive control, indomethacin. The 95 % ethanolic extract of Glycine was further fractionated using polyamide and silica gel column into 11 fractions. Blood thinning effects were observed in vitro by mixing the Glycine root extracts with human red blood cells. The addition of the 11 fractions of 95 % ethanolic extract of Glycine root increased the flow behavior index (n) from 0.68 to 0.72~0.78, and reduced fluid consistency (K) from 98 to 57~79 cP indicative of a change in flow behavior becoming closer to Newtonian type and a thinning consistency. The antioxidant activities of the 5 fractions eluted from silica gel column were higher than the 95 % ethanolic crude extract. The IC50 values were 15~70 % of the latter. The IC50 values of all fractions on DPPH free radical-scavenging activity were 2~21 times than the 95 % ethanolic crude extracts. The 11 fractions of the 95 % ethanolic extract showed inhibitory ability on LOX being 0.36~1.67 % of the NDGA (nordihydroguaiaretic acid), and 2.6~17.8 times of the 95 % ethanolic extract. The inhibitory activity of the fraction VIII was the highest among the 11 fractions. Fraction VIII was further separated using HPLC, and preliminarily identified using TLC sprayed with coloring agents, and UV-Vis spectrum. Isoflavone without glycoside was considered the bioactive component, which was further identified with NMR to be daidzein and its inhibitory ability on LOX was 1.4 and 23 times of the fraction VIII and the 95 % ethanolic extract.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0000000152
http://ntour.ntou.edu.tw/ir/handle/987654321/14520
Appears in Collections:[食品科學系] 博碩士論文

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