臺灣博碩士論文加值系統

二年生金門產一條根(闊葉大豆之主根部)樣品長度為12.0 ~ 14.3 cm。一條根之水、50 %乙醇以及95 %乙醇粗萃取物中，多酚類 (gallic acid equivalent) 含量分別為34.82、103.72以及88.54 mg/g，而三種溶劑中以95 %乙醇之萃取產率為最高(12.0%)，其次為水萃取之產率(10.1%)，50 %乙醇萃取產率最低(9.4%)。
對血色素催化亞麻油酸(linoleic acid)的抑制能力，以95 %乙醇粗萃取物之IC50最低 (4.63 μg/ml)，其次為50 %乙醇粗萃取物 (IC50=9.06 μg/ml)，水粗萃取物抑制脂肪酸氧化之能力為三者中最低(IC50=28.16 μg/ml)，一條根萃取溶劑之極性較低，其萃取物之抗氧化性較高。
掃除DPPH (α, α-diphenyl-β-picrylhydrazyl)自由基之抗氧化試驗中，發現三種粗萃取物皆具有掃除DPPH自由基之抗氧化特性，其掃除之能力依序為95 %乙醇粗萃取物 > 50 %乙醇粗萃取物 > 水粗萃取物，其結果與抑制脂肪酸自氧化之能力一致。
金門產一條根粗萃取物對於黃豆脂氧合(soybean lipoxygenase, LOX)皆具有抑制作用，產物與發光劑luminol產生之冷光，以95 %乙醇粗萃取物最高，水粗萃取物居末；三者皆可降低15-LOX催化花生四烯酸(eicosatetraenoic acid)形成之15-氫過氧化物(15- hydroperoxy eicosatetraenoic acid, 15-HpETE)之產量，其抑制率以95 %乙醇粗萃取物最高；三種粗萃取物皆具有螯合鐵離子之能力，可能會與LOX活性中心之鐵離子產生螯合，使酵素失活。
三種一條根粗萃取物對於人類血液流變性質之影響，根據冪次方程式σ=K۰γn ，其流體行為指標(fluid behavior index, n)皆為0.71，皆較未添加一條根萃取物之血液(n=0.65)更接近牛頓型流體(n=1)；而添加三種一條根萃取物黏稠指標(fluid consistency, K)皆為0.08，較控制組(K=0.13)低，代表一條根萃取物具有降低血液黏稠度之功能，並增加紅血球細胞膜之完整性。
一條根95%乙醇粗萃取物利用polyamide管柱及流動相水、10 %乙醇、30 %乙醇、50 %乙醇、70 %乙醇、95 %乙醇、氯仿-甲醇(1:1)、及甲醇分離，發現
50 %乙醇分離物在A254nm之純度為88.6 %，並經呈色試驗推測為黃酮類，在UV及可見光譜之吸光波峰類似epicatechin；此分離物具有抑制亞麻油酸自氧化、掃除DPPH之能力、及抑制LOX作用亞麻油酸產物與luminol反應之冷光之抗氧化特性，但不具螯合

I-Tiao-Gung, root of Glycine tomentella Hayata, used in this study was about 12.0 ~ 14.3 cm in length. The polyphenol content expressed as gallic acid equivalent extracted from I-Tiao-Gung using water, 50 % ethanol and 95 % ethanol as solvent were 34.82, 103.72 and 88.54 mg/g respectively. Extraction with 95 % ethanol gave the highest yield (12.0 %).
Inhibition of hemoglobin-induced linoleic acid (C18:2) oxidation by root extracts of I-Tiao-Gung was indicated by the IC50 being 4.63, 9.06, 28.16 μg/ml, for the 95 % ethanol, 50 % ethanol, water extract, decreased with increased polarity of the extraction solvent.
All three crude extracts had free radical-scavenging activity determined with DPPH. The capacity was similar with inhibition of lipid autoxidation in an order of 95 % ethanol extract > 50 % ethanol > water.
Inhibition on the soybean lipoxygenase catalyzed arachidonate oxidation was the highest with 95 % ethanol extract, and followed the same order in reducing 15-hydroperoxy eicosatetraenoic acid (15-HpETE) formation shown by HPLC, and chemiluminescence intensity using luminol. All three crude extracts chelated iron ion, while the lipoxygenase active site has Fe3+.
Blood thinning effects were observed in vitro by mixing the extract of I-Tiao-Gung with human red blood cells. The rheological data fitted the power law model, σ = K γ n, where σ =shear stress, γ = shear rate, K = fluid consistency, n =flow behavior index, with a correlation coefficient of R2 = 0.99 at a constant hematocrit of 44 %. The addition of extract resulted in a reduced K from 0.13 to 0.08, and an increased n from 0.65 to 0.71 indicative of a change in flow behavior closer to Newtonian type and a thinning consistency.
Separation of the crude 95 % ethanol extract of I-Tiao-Gung using polyamide column eluted by water, 10 % ethanol 30 % ethanol, 50 % ethanol, 70 % ethanol, 95 % ethanol, methanol-chloroform (1:1), and methanol as mobil phase, obtained one fraction eluted by 50 % ethanol showing a purity of 88.6 % as detected with A254nm. The fraction had color reactions and UV-Vis spectrum similar to those of epicatechin. It inhibited linolate autoxidation, and lipoxygenase activity, as well as DPPH free radical scavenging, but did not chelate iron.

中文摘要 …………………………………………………… I
英文摘要 ………………………………………………………… III
壹、前言 ………………………………………………………… 1
貳、文獻整理 ……………………………………………… 4
一、金門產一條根之藥用植物學考察及毒性 ………………… 4
二、多酚類化合物之分佈及定性 ……………………………… 6
三、抗氧化劑對於紅血球與血液黏度之影響 ………………… 7
四、脂氧合與血液黏度之關係 ……………………………… 9
五、脂氧合與心血管疾病的關係 …………………………… 10
六、5-脂氧合在動物體之生理功能 ………………………… 12
七、發炎及過敏反應之抑制 …………………………………… 14
八、12-lipoxygenase及生成物hepoxilin之生理功能………… 17
九、12-lipoxygenase之抑制作用 ……………………………… 19
參、材料與方法 ………………………………………………… 21
一、材料 ………………………………………………………… 21
二、分析方法 ……………………………………………… 21
1. 一條根萃取物之製備………………………………………… 21
2. 總多酚類之定量……………………………………………… 22
3. 對血色素催化亞麻油酸自氧化之抑制作用………………… 22
4. 脂氧合總活性測定………………………………………… 23
5. 逆向高效液相層析法測定脂氧合異構活性…………… 24
6. 血液黏度測定………………………………………………… 25
7. 血液滲透壓之測定…………………………………………… 25
8. 掃除自由基試驗……………………………………………… 25
9. 螯合鐵能力之測定…………………………………………… 26
10. 一條根粗萃取物之初步分離………………………………… 27
11. 酚類化合物之HPLC分析……………………………………… 28
12. 分離物之呈色反應………………………………………… 28
肆、結果與討論 ……………………………………………… 29
一、一條根萃取條件 …………………………………………… 29
1. 溶劑之選擇…………………………………………………… 29
2. 萃取次數及時間之選擇……………………………………… 30
二、一條根粗萃取物之功能特性 ……………………………… 30
1. 產率及多酚類之含量………………………………………… 30
2. 一條根粗萃取物對於脂肪酸自氧化作用之影響…………… 33
(1) 對亞麻油酸自氧化之影響 ……………………………………33
(2) 掃除DPPH自由基之能力 …………………………………… 35
3. 對脂氧合催化不飽和脂肪酸氧化之影響………………… 37
(1) 脂氧合總活性抑制………………………………………… 38
(2) 脂氧合異構之抑制 …………………………………… 40
4. 一條根粗萃取物對人血液特性之影響.……………………… 42
三、一條根萃取物多酚類之組成與抗氧化性…………………… 45
1. HPLC分離條件之確定………………………………………… 45
2.一條根分離物之多酚類含量及組成…………………………… 48
(1)水分離物 ……………………………………………………… 48
(2) 10 %乙醇分離物……………………………………………… 52
(3) 30 %乙醇分離物……………………………………………… 54
(4) 50 %乙醇分離物……………………………………………… 56
(5) 70 %乙醇分離物……………………………………………… 59
(6) 95 %乙醇分離物、氯仿-甲醇分離物、及甲醇分離物…… 60
3. 95%乙醇分離物經polyamide管柱層析分離物之抗氧化性… 61
(1) 對黃豆脂氧合作用亞麻油酸之影響 …………………… 61
(2) 對血色素催化脂質自氧化之影響…………………………… 64
伍、結論 ………………………………………………………… 66
陸、參考文獻 …………………………………………………… 68

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