臺灣博碩士論文加值系統

本研究以2004年二期作糙米（台稉9號）與綠豆(台南5號)、花生(台南11號)及青仁黑豆(台南3號)種子經適當發芽至芽長0.5-1.0 mm，配合超甜玉米（金蜜1號）粒為原料，首先探討合適的萃取法及萃取溶劑，繼而探討發芽對穀豆種子抗氧化力及機能性成份之影響，分析機能性成份與抗氧化力特性間關係，並以反應曲面法探討原料不同比例組合之最佳總抗氧化力配方及各原料對混合物之總抗氧化力影響，之後再探討最適配方之抗致突變性。結果如下：超音波加熱萃取所得萃取液之總抗氧化力較水浴加熱振盪萃取者為高；水萃液較95%乙醇萃取液之抗氧化力佳，後者又較正己烷者佳。發芽處理影響的探討則顯示豆類經發芽處理，其總抗氧化力皆會增加，而糙米則會降低；DPPH自由基清除能力則都會降低，亞鐵離子螯合能力在豆類中似以增加者居多；至於還原力則皆不受影響。另亦顯示各榖豆芽水萃液中總酚、類黃酮、異黃酮素、抗壞血酸及白黎蘆醇含量皆高於穀豆種子者，但在花青素部分，除黑豆種子水萃液含量高於發芽黑豆者外，其餘原料則均沒有差異。機能性成份與抗氧化力則有如下關係：（1）穀豆種子方面，發現總酚含量增加則總抗氧化力提高。另外總酚含量與DPPH自由基清除能力、亞鐵離子螯合能力及還原力皆成正相關。（2）穀豆芽則顯示其機能性成份與還原力皆呈顯著正相關。機能性成份中，維生素C與各種抗氧化力間都有很高的相關性。總酚含量與總抗氧化力、DPPH自由基清除能力及還原力都呈正相關。
在榖豆芽混合物最適配方的研究中則發現發芽綠豆、發芽花生及發芽糙米添加比例與混合物之總抗氧化能力成反比，發芽黑豆添加比例則與之成正比，但由於各成份間存在交感作用，使反應曲面呈複雜的曲線。在超甜玉米粒及發芽綠豆添加比例為0.00 %時，總抗氧化力最強之配方為發芽糙米：發芽黑豆：發芽花生＝23.5 %：73.2 %：3.3 %。最適配方的抗致突變性試驗顯示此配方並無毒性效應及致突變性，在TA98及TA100菌株系統中則皆有抗致突變效果，其中以TA100菌株系統抑制AFB1及NQNO致突變劑所引起的致突變性的效果較佳。

Seeds of Cereals and legumes, summed up to five kinds, were used as raw materials in this study. The seeds used, all produced in year of 2004, were Taiken-9 brown rice of second crop season, mung bean of Tainan no. 5, peanut of Tainan no.11, black soybean with green kernel of Tainan no.3 and sweet corn of King-me no. 1. All seeds, except sweet corn, were soaked in water for a proper time, and then pick it on cotton sheet for germination. The germination terminated while the sprouts grew to 0.5-1.0 mm in length. The four kinds of germinated seeds and sweet corn seed then used to prepare nutrient adjuncts. The exploration of suitable extraction method and solvent were done at first, then, the effect of germination on the anti-oxidative activities and contents of functional components and the relationships between the anti-oxidative activities and functional components were investigated, and following the optimal formulated mixture with peak anti-oxidative activity was sought out. The mixture selected was then undergone a anti- mutagenictiy test to guarantee its safety. The results showed that most of four anti-oxidative activities, i.e., Total anti-oxidative activity, the DPPH radical scavenging power, the ferrous ion chelating power and the reducing power, were higher in the water extract than that of 95% ethanol extract and normal hexane extract either using ultrasound or shaking water bath extraction. As to the extraction method, ultrasound method showed higher anti-oxidative power in the extract than that of the extract of water bath combined with shaking. The investigation on the effect of germination on the anti-oxidative power of seeds showed that germination would cause increase in total anti-oxidative activity in legumes’ seeds, but decrease in brown rice. The DPPH scavenging power decreased for all kinds of seeds. All kinds of legumes’ seeds showed an increase in the ferrous ion chelating power after germination process, though there was no significant difference between reducing power of germinated and non-germinated seeds. The results also showed that the amount of all the functional components, except for anthocyanin, were higher in germinated seeds than that of the non-germinated seeds. The germinated black soybean seed showed a lower amount of anthocyanin, dut to the high water-soluble property of anthocyanin, than that of the non-germinated ones. It was also showed that a positive correlation between the amount of total polyphenols and all the four anti-oxidative activities either in non-germinated or germinated seeds. In addition, there were high correlation between the amount of vitamin C and all the four anti-oxidative properties. In the exploration of optimal formulation of nutriention adjunct, the result showed that total anti-oxidative activity raised while the level of germinated black soybean increased. It was suggested that there were interactions between different seeds on the total anti-oxidative activity, so it was better not to use single raw material. The optimal formulation with the highest total anti-oxidative activity for nutriention adjunct while the addition ratio of the five raw materials was as following: germinated brown rice : germinated black soybean: germinated peanut =23.5%: 73.2%: 3.3% while the addition level was set to be zero for sweet corn and =germinated mung bean.

目錄
中文摘要……………………………………………………………………………………………i
英文摘要…………………………………………………………………………………………iii
壹、前言……………………………………………………………………………………………1
貳、文獻回顧………………………………………………………………………………………3
ㄧ、五種穀、豆類原料簡介………………………………………………………………………3
1.1黑豆……………………………………………………………………………………………3
1.1.1 黑豆簡介…………………………………………………………………………………3
1.1.2 黑豆的功用和藥性………………………………………………………………………3
1.1.3 黑豆的營養及機能性……………………………………………………………………3
1.1.4 黑豆芽之研究……………………………………………………………………………4
1.2 綠豆…………………………………………………………………………………………4
1.2.1 綠豆簡介…………………………………………………………………………………4
1.2.2 綠豆的功用和藥性………………………………………………………………………5
1.2.3 綠豆的營養及機能性……………………………………………………………………5
1.2.4 綠豆芽之研究……………………………………………………………………………5
1.3 玉米…………………………………………………………………………………………6
1.3.1玉米簡介…………………………………………………………………………………6
1.3.2 玉米的功用和藥性………………………………………………………………………7
1.3.3 玉米的營養及機能性……………………………………………………………………7
1.4 花生…………………………………………………………………………………………8
1.4.1 花生之簡介………………………………………………………………………………8
1.4.2 花生的功用和藥性………………………………………………………………………8
1.4.3 花生的營養及機能性……………………………………………………………………8
1.4.4 花生芽之研究……………………………………………………………………………9
1.5糙米…………………………………………………………………………………………9
1.5.1糙米簡介…………………………………………………………………………………9
1.5.2 糙米的營養及機能性……………………………………………………………………10
1.5.3 發芽糙米之研究…………………………………………………………………………10
二、自由基與活性氧的定義及特性……………………………………………………………11
2.1自由基………………………………………………………………………………………11
2.2活性氧………………………………………………………………………………………11
2.3自由基的來源………………………………………………………………………………11
三、自由基與活性氧的清除及自由基性損傷的修復…………………………………………12
第一道防線：細胞色素氧化酶使氧直接還原成水……………………………………………13
第二道防線：抗氧化酶…………………………………………………………………………13
第三道防線：抗氧化劑（清除劑）……………………………………………………………14
（1）維生素C……………………………………………………………………………………15
（2）類胡蘿蔔素………………………………………………………………………………15
（3）酚的衍生…………………………………………………………………………………16
（4）維生素E…………………………………………………………………………………18
（5）間接清除劑………………………………………………………………………………18
第四道防線：自由基性損傷的修復……………………………………………………………20
（a）蛋白質的降解和修復………………………………………………………………………20
（b）DNA的降解和修復…………………………………………………………………………21
第五道防線：凋亡……….………………………………………………………………………21
三、抗氧化劑間相互作用及其複雜性…………………………………………………………21
3.1互相配合……………………………………………………………………………………21
3.2互相保護……………………………………………………………………………………23
3.3互相互補……………………………………………………………………………………23
四、安氏試驗法…………………………………………………………………………………23
4.1測試菌株……………………………………………………………………………………24
4.2.藥物轉化酵素S9（鼠肝混合物）………………………………………………………25
五、食物中的致突變物/致癌物………………………………………………………………26
5.1天然存在食物中的植物鹼及微生物或植物毒素…………………………………………26
5.2食物經由儲存、加工或烹調等過程所產生的化合物……………………………………27
5.3食物受污染或殘留的殺蟲劑及人工合成的化合物………………………………………28
六、食品中的抗致突變物及抗致癌物…………………………………………………………28
七、複方與相乘性………………………………………………………………………………29
参、材料與方法…………………………………………………………………………………31
一、試驗架構……………………………………………………………………………………31
二、試驗材料……………………………………………………………………………………34
三、化學藥品與溶劑……………………………………………………………………………34
（一）化學藥品…………………………………………………………………………………34
（二）溶劑………………………………………………………………………………………35
四、儀器設備……………………………………………………………………………………35
五、試驗方法……………………………………………………………………………………36
5.1穀、豆芽凍乾粉末製備……………………………………………………………………36
5.1.1 穀、豆芽原料製備………………………………………………………………36
5.1.1.1 發芽操作……………………………………………………………………36
5.1.1.1.1糙米之發芽操作…………………………………………………………37
5.1.1.1.2花生、綠豆及黑豆種子之發芽操作……………………………………37
5.1.1.1.2.1花生、綠豆及黑豆種子之浸泡溫度及時間的決定………………37
5.1.1.1.2.2花生、綠豆及青仁黑豆種子催芽時間的決定……………………38
5.1.2穀豆芽凍乾粉末製備……………………………………………………………38
5.2 萃取溶劑及萃取法選取…………………………………………………………………38
5.3 穀豆芽各種抗氧化力評估………………………………………………………………40
5.3.1 萃取液製備………………………………………………………………………40
5.3.2 各種抗氧化力評估………………………………………………………………40
5.3.2.1α,α-diphenyl-β-picrylhydrazyl自由基清除能力測定……………40
5.3.2.2螯合亞鐵離子能力測定……………………………………………………41
5.3.2.3還原力測定……………………………………………………………………41
5.4各穀、豆芽機能性成分含量之定量………………………………………………………42
5.4.1類黃酮化合物之定量……………………………………………………………42
5.4.2總酚類化合物之定量……………………………………………………………43
5.4.3花青素之定量……………………………………………………………………43
5.4.4抗壞血酸之定量…………………………………………………………………43
5.4.5異黃酮素及白黎蘆醇之定量……………………………………………………44
5.5穀、豆芽最適配方探討…………………………………………………………………46
5.6穀豆芽混合物最適配方之抗致突變性試驗……………………………………………46
5.6.1毒性試驗………………………………………………………………………………46
5.6.2 致突變性分析………………………………………………………………………49
5.6.3 抗致突變性分析……………………………………………………………………49
肆、結果與討論………………………………………………………………………………51
一、花生、綠豆及黑豆種子之發芽操作……………………………………………………51
1.1花生、綠豆及黑豆種子之浸泡溫度及時間的決定……………………………………51
1.2花生、綠豆及青仁黑豆種子催芽時間的決定………………………………………54
二、萃取溶劑及萃取法選取…………………………………………………………………54
三、各穀豆種子及其芽的抗氧化能力比較及發芽對各穀豆種子抗氧化力的影響………61
3.1 總抗氧化力比較…………………………………………………………………………61
3.1.1各穀豆種子總抗氧化力比較……………………………………………………………61
3.1.2各穀豆芽總抗氧化力比較………………………………………………………………63
3.1.3發芽對各穀豆種子總抗氧化力的影響…………………………………………………63
3.2 DPPH自由基清除能力比較………………………………………………………………63
3.2.1各穀豆種子DPPH自由基清除能力比較…………………………………………………63
3.2.2各穀豆芽DPPH自由基清除能力比較……………………………………………………65
3.2.3發芽對各穀豆種子DPPH自由基清除能力的影響………………………………………66
3.3亞鐵離子螯合能力比較……………………………………………………………………66
3.3.1各穀豆種子亞鐵離子螯合能力比較……………………………………………………66
3.3.2各穀豆芽亞鐵離子螯合能力比較………………………………………………………68
3.3.3發芽對穀豆種子亞鐵離子螯合能力的影響……………………………………………69
3.4 還原能力比較………………………………………………………………………………69
3.4.1各穀豆種子還原力比較…………………………………………………………………69
3.4.2各穀豆芽還原能力比較…………………………………………………………………71
3.4.3發芽對穀豆種子還原力的影響…………………………………………………………71
3.5穀豆種子及其芽之抗氧化力綜合比較……………………………………………………71
3.5.1穀豆種子之抗氧化力綜合比較…………………………………………………………71
3.5.2穀豆芽之抗氧化力綜合比較……………………………………………………………72
3.5.3發芽對穀豆種子抗氧化力之綜合影響…………………………………………………72
四、穀豆種子及其芽之機能性成份含量比較及發芽對其影響………………………………74
4.1穀豆種子機能性成份含量比較……………………………………………………………74
4.2穀豆芽機能性成份含量比較………………………………………………………………74
4.3發芽對機能性成份含量的影響……………………………………………………………76
五、機能性成份與抗氧化力特性間關係探討…………………………………………………76
5.1穀豆種子機能性成份與抗氧化力特性間關係探討………………………………………76
5.2穀豆芽機能性成份與抗氧化力特性間關係探討…………………………………………77
六、穀豆芽最適配方探討………………………………………………………………………82
七、穀豆芽混合物最適配方之抗致突變性試驗………………………………………………92
7.1毒性試驗……………………………………………………………………………………92
7.2致突變性試驗………………………………………………………………………………92
7.3抗致突變性試驗……………………………………………………………………………93
伍、結論………………………………………………………………………………………99
陸、參考文獻…………………………………………………………………………………101
表 次
表一、主要自由基來源…………………………………………………………………………12
表二、穀、豆芽營養補助品最適配方研製操作過程中，各種不同混合比例組合之實驗設計…………………………………………………………………………………………………48
表三、糙米、黑豆、花生及綠豆發芽時間與芽長及發芽率的關係…………………………53
表四、各種穀、豆芽之水和乙醇萃取物之抗氧化力比較……………………………………60
表五、發芽對穀豆種子抗氧化力的綜合影響…………………………………………………73
表六、穀豆種子及穀豆芽的機能性成份含量及抗氧化能力…………………………………75
表七、穀豆類及超甜玉米種子的機能性成份含量及抗氧化能力之相關係數矩陣…………79
表八、穀豆種子的機能性成份含量及抗氧化能力之相關係數矩陣…………………………80
表九、穀豆芽的機能性成份含量及抗氧化能力之相關係數矩陣……………………………81
表十、最適配方探討試驗中，五種穀豆芽各種不同混合比例組合及各種回應變數值……83
表十一、五種穀豆芽的各種不同混合處理之機能性成份與抗氧化力之相關係數矩陣……85
表十二、穀豆芽最適配方研製過程中，總抗氧化力順序模式與欠合性分析………………86
表十三、穀豆芽最適配方研製過程中，總抗氧化力之變方分析……………………………87
表十四、穀豆芽最適配方研製過程中，總抗氧化力與各因子間之迴歸方程式……………88
表十五、穀豆芽混合物最適配方組成…………………………………………………………91
表十六、綜合穀、豆芽營養補助品最適配方萃取物對鼠傷寒沙門桿菌TA98及TA100菌株之毒性試驗 (± S9)………………………………………………………………………………95
表十七、綜合穀、豆芽營養補助品最適配方萃取物對鼠傷寒沙門桿菌TA98及TA100菌株之致突變(± S9)……………………………………………………………………………………96
表十八、綜合穀、豆芽營養補助品最適配方萃取物對4-nitro-quinoline-N-oxide (NQNO) 在鼠傷寒沙門桿菌TA98及TA100菌株之抗致突變性測定………………………….97
表十九、綜合穀、豆芽營養補助品最適配方萃取物對aflatoxin B1（AFB1）在鼠傷寒沙門桿菌TA98及TA100菌株之抗致突變測定……………………………………………………98
圖 次
圖一、類胡蘿蔔素的化學結構…………………………………………………………………17
圖二、類黃酮化合物的化學結構………………………………………………………………19
圖三、維生素E、C和GSH的協同清除自由基作用……………………………………………22
圖四、抗氧化酶間的互相保護作用……………………………………………………………23
圖五、機能性綜合穀豆芽營養補助品最適配方之試驗架構…………………………………31
圖六、五種穀豆芽各種抗氧化活性及機能性成分含量評估之流程圖………………………32
圖七、穀豆芽混合物最適配方之抗致突變性評估實驗流程圖………………………………33
圖八、花生、綠豆及黑豆浸泡時之吸水曲線…………………………………………………52
圖九（A）、不同萃取法和萃取溶劑所得發芽糙米萃取液之總抗氧化力之比較……………55
圖九（B）、不同萃取法和萃取溶劑所得超甜玉米粒萃取液之總抗氧化力之比較…………56
圖九（C）、不同萃取法和萃取溶劑所得發芽黑豆萃取液之總抗氧化力之比較……………57
圖九（D）、不同萃取法和萃取溶劑所得發芽花生萃取液之總抗氧化力之比較……………58
圖九（E）、不同萃取法和萃取溶劑所得發芽綠豆萃取液之總抗氧化力之比較……………59
圖十、五種穀豆類種子及穀豆芽水萃取液之總抗氧化力……………………………………62
圖十一、五種穀、豆類種子及穀豆芽水萃取液之DPPH自由基清除能力……………………64
圖十二、五種穀、豆類種子及穀豆芽水萃取液之亞鐵離子螯合能力………………………67
圖十三、五種穀、豆類種子及穀豆芽水萃取液之還原力……………………………………70
圖十四、在超甜玉米添加量為0，綠豆添加量變動為（a）0（b）6.4條件下，發芽黑豆、花生及發芽糙米添加量對總抗氧化力之影響………………………………………………89
圖十五、在超甜玉米添加量為0，綠豆添加量變動為（c）12.8（d）19.2（e）25條件下，發芽黑豆、花生及發芽糙米添加量對總抗氧化力之影響……………………………90

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