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研究生:劉俊毅
研究生(外文):Liou, Jiunyi
論文名稱:金柑萃取物抑制鯖魚油氧化之活性及其有效成分
論文名稱(外文):Inhibitory Activity Of Kumquat Extracts On Lipid Oxidation Of Mackerel And It’s Effective Compounds
指導教授:駱錫能駱錫能引用關係
指導教授(外文):Lou, Shyineng
口試委員:邱思魁盧義發張永鍾邱一鳴
口試委員(外文):Chiou,TzekueiLu, YifaChang, YungchungChiou, Yiming
口試日期:2012-07-11
學位類別:碩士
校院名稱:國立宜蘭大學
系所名稱:食品科學系碩士班
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:116
中文關鍵詞:金柑類黃酮鯖魚油脂質氧化Acacetin 7-O-neohesperidoside
外文關鍵詞:Kumquatflavonoidsmackerel lipidslipid oxidationAcacetin 7-O-neohesperidoside
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本研究以未成熟長實金柑為材料,分別以熱水、乙醇及乙酸乙脂萃取後,添加至鯖魚油中,探討萃取物之抑制脂質氧化能力,再以半製備C18管柱劃分收集,確認有效抑制脂質氧化之劃分物,並進一步分析抗氧化模式及鑑定其有效成分。
結果顯示,熱水及乙醇萃取物的清除DPPH自由基能力高於乙酸乙脂萃取物,其有效DPPH清除率分別為7.51±0.67和6.98±0.30 % / mg / mL,相當於158.96 ±5.39和119.93 ±3.81 M trolox。將乙醇萃取物添加至鯖魚油中進行加熱貯藏試驗,發現添加2000 ppm金柑乙醇萃取物可以有效延緩氧化誘導期至第5天,並能減少共軛雙烯化合物之生成,且可延緩EPA及DHA的氧化裂解。
以半製備C18管柱分離金柑乙醇萃取物為六個劃分物,再添加至鯖魚油中進行加熱貯藏試驗,得知劃分物I能夠延緩氧化誘導期至第3天,劃分物V及VI則有抑制共軛雙烯生成的能力,顯示抗氧化有效成分可能與劃分物I、V及VI等有關。
經抗氧化測試結果顯示,劃分物I具有較佳的清除DPPH自由基及清除超氧陰離子能力,其值分別為22.79±6.10 %及46.04±19.91 %。此外,劃分物I、V及VI皆具有氧自由基吸收能力(ORAC),其中以劃分物V的吸收能力最佳,其值相當於1785±9 M trolox。混合不同比例劃分物I、劃分物V、金柑乙醇萃取物及BHA等進行ORAC測試,發現金柑乙醇萃取物與BHA以667 ppm+133 ppm混合時有最佳提高ORAC的效果,其提高率達419%,次佳則為劃分物I與BHA以667 ppm+67 ppm混合,可提高20.6% ORAC的效果。
分析劃分物I和V的組成,顯示劃分物I含有甚多不同化合物,而劃分物V 則僅有一主要波峰,經收集純化後,以UV scan進行掃描,劃分物V之UV max為 267與332 nm,再經質譜分析後得知分子量為592 dalton,主要裂解離子MS1為447 [M+H–146]+ 與MS2 285 [M+H–146–162]+,經與文獻比對後,初步鑑定結構為 Acacetin 7-O-neohesperidoside,在未成熟金柑中含量達0.56 g / 100 g d.w.。

In this study immature kumquat was extracted by hot water, ethanol, and ethyl acetate. The obtained extracts were added in oil of mackerel, separately, to evaluate the inhibitory activity on lipid oxidation under heating condition. In order to confirm the effective compounds, a semi-preparative C18 column was used to separate and fractionate the target substances. Furthermore, the inhibitory model of antioxidant and identification of the effective compounds were also investigated.
The results exhibited that DPPH free radical scavenging effect of extracts from hot water and ethanol extracts were better than ethyl acetate extract, while the scavenging potency were 7.51±0.67 and 6.98±0.30 % / mg / mL, respectively, which also equal to 158.96 ±5.39 and 119.93 ±3.81 M trolox. After addition of ethanol extract of kumquat into oil of mackerel, evaluation of inhibitory activity against lipid oxidation was carried out under heating storage. The results showed that 2000 ppm ethanol extract could retard the induction time to 5 days against lipid oxidation. Besides, it could decrease the formation of conjugated diene substances and also postpone the degradation of EPA and DHA, effectively. The ethanol extract was separated to six fractions via semi-preparative C18 column, and each fraction was added to oil of mackerel for the accelerated storage test. Fraction I could delay the oxidation induction time to third day, while fraction V and fraction VI could inhibit the formation of conjugated diene. These hinted that the compounds in fraction I, V and VI might relate to the antioxidant activity.
The results of antioxidant activity test revealed that fraction I had better DPPH free radical and superoxide anion radical scavenging activity, with values of 22.79 ± 6.10 % and 46.04 ± 19.91 %, respectively. Moreover, the highest ORAC(oxygen-radical absorbance capacity)of fraction V with 1785±9 M trolox was also observed. We used different mixtures of kumquat ethanol extract(K), fraction I(I), fraction V(V), and BHA in the ORAC assay. A increasing effect of ORAC assay to 419% was observed with the ratio of K:BHA=5:1. A increase of 20.6% ORAC of I:B=10:1 was also found.
In fraction I, many compounds existed, however, there was only a major compound in fraction V. The compound was detected by UV scan and the maximum absorptions were 267 and 332 nm. The molecule weight was certified as 592 dalton. The main breaking ion for MS1 was 447 [M+H–146]+ and for MS2 was 285 [M+H–146–162]+. It was identified as acacetin 7-O-neohesperidoside in comparing with literature data. The level of acacetin 7-O-neo- hesperidoside in immature kumquat was 0.56 g / 100 g d.w.

摘要 I
Abstract III
目錄 V
表目錄 IX
圖目錄 X
壹、前言 1
貳、文獻整理 3
一、金柑 3
(一)品種與生長習性 3
(二)產量及分布概況 5
(三)組成成份與生理功能 5
二、柑橘類水果中抗氧化相關成分 10
(一)類黃酮 10
(二)酚酸 22
三、天然萃取物對脂質氧化之影響 24
四、鯖魚魚油的特性 26
(一)鯖魚 26
(二)脂肪酸組成 28
(三)安定性 30
參、實驗架構 32
一、不同溶劑萃取之抗氧化能力試驗 32
二、金柑萃取物抑制鯖魚油脂質氧化試驗 32
三、金柑劃分物抑制鯖魚油脂質氧化試驗 33
四、劃分物抗氧化模式判定及加乘效果試驗 33
肆、材料與方法 34
一、實驗材料 34
(一)金柑 34
(二)鯖魚 34
二、藥品及器材 34
(一)藥品 34
(二)器材 35
三、方法 36
(一)樣品製備 36
(二)貯藏試驗 37
(三)油脂氧化指標測定 38
(四)脂肪酸組成分析 39
(五)抗氧化活性物質之半製備管柱劃分收集 41
(六)抗氧化模式測定 42
(七)有效成分組成分析 44
(八)質譜儀鑑定條件 47
四、統計分析 47
伍、結果與討論 48
一、金柑不同溶劑萃取物之抗氧化能力 48
二、添加金柑乙醇萃取物抑制鯖魚油脂氧化之效應 49
(一)貯藏中油脂氧化指標的變化 50
(二)貯藏中脂肪酸組成之變化 54
三、金柑乙醇萃取物之劃分收集及其抗氧化能力評估 58
(一)C18半製備管柱劃分收集 58
(二)劃分物抑制鯖魚油脂氧化之能力 58
(三)劃分物抗氧化模式初步判定 61
四、劃分物抗氧化加乘效應之探討 63
五、相關有效成分之探討 64
陸、結論 66
柒、參考文獻 67
捌、表 83
玖、圖 91

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