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研究生:黃子軒
研究生(外文):Zih-Syuan Huang
論文名稱:高粱酒糟萃取液之製備及其有機酸成分與胞外生物活性之評估
論文名稱(外文):Preparation of extracts from sorghum distillery residues and investigation of their organic acid composition and in vitro bioactivities
指導教授:陳錦樹陳錦樹引用關係
口試委員:蘇南維林俊杰蔡碩文
口試日期:2016-07-22
學位類別:碩士
校院名稱:國立中興大學
系所名稱:食品暨應用生物科技學系所
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:2016
畢業學年度:104
語文別:中文
論文頁數:127
中文關鍵詞:高粱酒糟有機酸抗氧化能力抑制α-澱粉酶活性抑制酪胺酸酶活性
外文關鍵詞:sorghum distillery residuesorganic acidantioxidant abilityinhibition of α-amylase activityinhibition of tyrosinase activity
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現今台灣菸酒公司高粱酒之釀造,皆以紅、白高粱混合作為主原料,經由二釀二蒸或三釀三蒸程序取得高粱酒液,剩餘之高粱酒糟即為加工副產物,早期常供作飼料用,但近年來逐漸被商業混合飼料取代,導致高粱酒糟利用率降低。
然而,高粱中富含許多具生物活性之機能性成分,於發酵及蒸餾製程中,將逐步釋出並存留於高粱酒糟中,如粗蛋白、胺基酸、維生素、膳食纖維、醇類、酚類、酸類等具應用潛力之化合物,若將高粱酒糟棄置甚為可惜。本研究使用由嘉義酒廠所提供之第一、第二及第三次發酵蒸餾後高粱酒糟作為試驗樣品,藉由去離子水及50%、60%、70%、80%(v/v)乙醇水溶液進行萃取條件之探討,並分析高粱酒糟萃取液中有機酸成分及含量、總多酚含量、總類黃酮含量、蛋白質與胜肽含量,以及探討抗氧化能力、抑制α-澱粉酶活性、抑制酪胺酸酶活性等三項生物活性,期能提升高粱酒糟之再利用價值。
上述三種不同高粱酒糟基本成分分析結果顯示,灰分、粗脂肪、粗蛋白及總膳食纖維上,皆以第三次發酵蒸餾後高粱酒糟具最高含量。經pH值、總酸含量、HPLC、總多酚及總類黃酮含量、抗氧化活性等測定後,初步得知80%乙醇萃取液中含醋酸、乳酸、阿魏酸、對-香豆酸、咖啡酸等有機酸,且含量為所有萃取組合中最高,於總多酚、總類黃酮含量及抗氧化活性亦為最佳,故選定萃取條件:第三次發酵蒸餾後高粱酒糟作為萃取樣品,使用80%乙醇溶液,以固液比1:5 (w/v, wet basis)、溫度60℃,於震盪水浴槽中以120 rpm萃取2小時,作為實驗探討之主軸。
另外,若於萃取前增加預處理:接種Aspergillus oryzae NCH 42及市售商業半纖維素酶,皆能有效提升萃取效果;而在pH值、總酸含量、總多酚含量、總類黃酮含量、DPPH自由基清除率、螯合亞鐵離子能力、還原力、超氧陰離子清除率、抑制α-澱粉酶及酪胺酸酶活性等測定上,大致皆以市售商業半纖維素酶預處理組高於接種Aspergillus oryzae NCH 42預處理組;而以高粱酒糟粉末萃取組再次之,且與原先之濕高粱酒糟萃取液較無明顯差異。

Nowadays, the leaders of sorghum liquor production in Taiwan are Kinmen Distillery, Lungtien Distillery and Chiayi Distillery, which uses a mixture of red and white sorghum as the primary ingredients and through two or three cycles of fermentation accompanied by distillation, sorghum distillery residue (SDR) is produced after processing along with the remainder as a by-product. In the early days, the by-product has be used as animal feed before the rise of commercial feed, and because of this, the usage of sorghum has decreased.
However, the functional ingredients found in sorghum such as crude protein, amino acid, vitamin, dietary fiber, alcohol, polyphenol, and acids will be continuously released and retained in the sorghum distillery residues while fermenting and distilling. Considering the beneficial effect of compounds, the sorghum distillery residues could be an added value and discarding it would be considered a waste.
In this study, the sample is composed of the sorghum distillery residues processed after three stages of fermentation and distillation. The samples were treated with deionized water and 50%, 60%, 70%, 80% ethanol during extraction, and the organic content, total polyphenol content, total flavonoid content, protein content and peptide content were analyzed. Furthermore, investigation of antioxidant capacity as well as inhibition ability of both amylase and tyrosinase were carried out, in hope of making the most use of sorghum distillery residues.
Based on the ingredient analysis of three different samples, the sorghum distillery residues after third fermentation and distillation showed the highest amount of ash, crude fat, crude protein, and total dietary fiber. In addition, through the measurement of pH value, total acidity, HPLC, total polyphenol content, total flavonoid content and antioxidant activity, the result indicated that the extract contains acetic acid, lactic acid , ferulic acid, p-coumaric acid, and caffeic acid. According to the preliminary result mentioned above, the extraction condition below was established for further investigation : the sorghum distillery residues after third fermentation and distillation was used as sample and extracted with 80% ethanol, at 60℃ in a water bath shaker under 120 rpm for 2 hours (SDRC).
There were three kinds of pretreatment: using sorghum distillery residues powders(SDRP), Aspergillus oryzae NCH 42(SDRA), and commercial hemicellulase(SDRH), used in the subsequent experiment. Based on the measurement of pH value, total acidity, total polyphenol content, total flavonoid content, DPPH radical scavenging activity, ferrous ions chelating effect, reducing power, superoxide anion scavenging activity, inhibition of α-amylase activity , and inhibition of tyrosinase activity,the result indicated that SDRH>SDRA>SDRP>SDRC.

摘要 i
Abstract ii
目次 iv
表目次 vii
圖目次 viii
第一章、前言 1
第二章、文獻回顧 2
一、高粱之介紹 2
(一)高粱之分類 2
(二)高粱之結構 5
(三)高粱之成分組成及生理活性 5
二、高粱酒之介紹 10
(一)高粱酒之簡介 10
(二)高粱酒之製程 11
三、高粱酒糟之介紹 15
(一)高粱酒糟之成分組成 15
(二)高粱酒糟之有機酸 15
(三)高粱酒糟之酚酸 18
(四)高粱酒糟之應用 21
四、抗氧化之介紹 24
(一)氧化與自由基生成 24
(二)抗氧化機制 27
(三)抗氧化系統及物質 28
五、抑制α-澱粉酶活性之介紹 29
(一)糖尿病之簡介 29
(二)糖尿病之分類 29
(三)糖尿病之用藥與治療 30
六、抑制酪胺酸酶活性之介紹 32
(一)皮膚之簡介 32
(二)皮膚黑色素生合成機制 34
(三)抑制黑色素生合成及天然美白成分 36
七、研究動機 36
第三章、材料與方法 38
一、實驗材料 38
(一)實驗原料 38
(二)酵素 38
(三)菌種 38
(四)化學藥劑 38
二、儀器設備 39
三、套裝軟體 40
四、實驗方法 41
(一)實驗大綱 41
(二)實驗架構 42
(三)高粱酒糟前處理 43
(四)高粱酒糟基本成分分析 43
(五)濕高粱酒糟萃取液之製備 46
(六)高粱酒糟萃取液之有機酸分析 46
(七)高粱酒糟萃取液之蛋白質含量分析 48
(八)高粱酒糟萃取液之胜肽含量分析 49
(九)高粱酒糟萃取液之抗氧化分析 49
(十)高粱酒糟萃取液之抑制酵素活性分析 51
第四章、結果與討論 53
一、高粱酒糟之基本成分分析 53
二、高粱酒糟萃取液之萃取條件初步探討 57
(一)乙醇濃度之探討 59
(二)萃取時間之探討 68
(三)萃取溫度及高粱酒糟之探討 73
三、高粱酒糟萃取液之有機酸成分及含量 80
(一)離子層析儀分析 80
(二)高效液相串聯層析質譜儀分析 80
四、高粱酒糟萃取方法之改良 83
(一)以高粱酒糟粉末萃取組 84
(二)接種Aspergillus oryzae NCH 42預處理組 84
(三)市售商業半纖維素酶預處理組 84
(四)不同預處理方式之pH值及總酸含量探討 85
(五)不同預處理方式之總多酚及總類黃酮含量探討 89
(六)不同預處理方式之蛋白質及胜肽含量探討 93
(七)不同預處理方式之抗氧化分析探討 97
(八)不同預處理方式之抑制α-澱粉酶活性分析探討 103
(九)不同預處理方式之抑制酪胺酸酶活性分析探討 105
第五章、結論 107
一、高粱酒糟之基本成分分析 107
二、高粱酒糟萃取液之萃取條件初步探討 107
三、高粱酒糟萃取液之有機酸成分及含量 107
四、高粱酒糟萃取方法之改良 108
第六章、未來展望 109
一、改善最適萃取條件及處理 109
二、HPLC分析 109
三、探討高粱酒糟萃取液之生物活性及應用 109
第七章、參考文獻 110
附錄一、不同高粱萃取液之萃取率 127

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