臺灣博碩士論文加值系統

抗性澱粉(RS)主要被分類成四種形式：RS1、RS2、RS3及RS4。其中最常見的為RS3，主要是澱粉經由糊化與貯存程序所產生之回凝澱粉，可經由加工取得；RS4則是經由化學修飾後產生的修飾澱粉。抗性澱粉依澱粉來源、結構差異、製備或處理條件不同，其物理化學特性皆不同，因此進行修飾反應的難易程度或產品品質也不同。本研究旨在探討由糊化與貯存程序所產生之回凝澱粉RS3，及預糊化澱粉經低分子量羧酸鈉酯化後之修飾澱粉，其結構與特性上之異同。使用方法除了測定其酵素消化率、吸水性、水溶性、直鏈澱粉複合指數、取代度等一般物理特性外，進一步利用示差掃描熱分析（DSC）、快速黏度測定儀（RVA）及掃描式電子顯微鏡（SEM）、質地剖面分析 (TPA) 等技術，分析其糊化、回凝、膠體結構。而傅立葉轉換紅外光譜儀 (FTIR) 可用以追尋RS3與RS4在酯鍵數量上的差異。
抵抗消化酵素之作用與對熱穩定，為符合抗性澱粉應用之兩大要素，而製作抗性澱粉的方式，採用水浴加熱與高壓殺菌釜，將生綠豆澱粉完全糊化後，於4℃貯存使其回凝，形成穩定的雙股螺旋直鏈澱粉及增加結晶度 (RS3)；或使用已完全糊化之澱粉樣品製備低分子羧酸鹽酯化澱粉 (RS4)，使之生成酯化鍵交聯以加強分子間的結合作用，進而對酵素產生抗性並具熱穩定。分析物理處理所得之回凝樣品，和經糊化處理後再進行酯化所得修飾澱粉之抗性澱粉含量，結果發現化學酯化生成抗性澱粉較物理回凝處理來的高，其中抗性澱粉製作又以單用檸檬酸鹽酯
化效果最佳，且物理回凝處理與化學修飾並用，較單獨使用化學修飾效果佳。吸水性指標與水溶性指標測定的結果顯示，澱粉經糊化回凝處理後，吸水性與水溶性指標隨貯藏時間延長而有下降之趨勢，表示其回凝程度增加，而經檸檬酸鹽酯化之澱粉，吸水性與水溶性指標隨檸檬酸鹽濃度與反應溫度提高而上升，推測其原因為親水性酯鍵的增加。示差掃描熱分析發現，生的綠豆澱粉其糊化溫度大約在67℃，而熱焓值為13.7 J/g，澱粉經糊化後因晶體遭受破壞，使得波峰消失，再經貯藏回凝或檸檬酸鹽酯化後，糊化吸收峰向高溫度方向移動，熱焓值有略微上升的趨勢，表示對熱之穩定性增加。快速黏度測定結果顯示，生澱粉的糊化溫度與示差掃描熱分析所得結果相近，且經糊化回凝處理之澱粉，回升黏度隨貯存時間延長而增加，但連續黏度極低，與原來生澱粉的特性完全不同。綠豆澱粉膠經掃描式電子顯微鏡觀察，証實澱粉糊化後，膠體網狀結構緊密度較差，隨貯存時間延長網狀結構變得更加緻密，而觀察檸檬酸鹽酯化澱粉，隨檸檬酸鹽濃度及反應溫度提高，網狀結構細密度增加至幾不可見。檸檬酸鹽酯化澱粉經傅立葉轉換紅外光譜分析，結果顯示隨羧酸鹽濃度及反應溫度提高，在1728 cm-1有一酯鍵吸收峰更加明顯，證實酯化澱粉確實有酯鍵產生而對酵素產生抗性。
綜合以上結果，經由物理性回凝所產生的RS3，與經由化學酯化所產生的RS4，在物化特性及光譜結構分析上，都存在有一些差異，而這也許對兩者在食品加工利用上有重要影響。

Resistant starches have mainly been categorized into 4 types: RS1, RS2, RS3, and RS4. RS3, generated via starch gelatinization followed by retrogradation, is the most common RS from processing. RS4 is the modified starch through chemical modification. The physicochemical characteristics of RS will depend on the source, structural difference, preparation, and treating condition of starch investigated. This study aims to reveal the differences in structures and characteristics, between gelatinized-retrograded RS3, and RS4 which originated from pregelatinized starch esterified with low molecular weight carboxylic acids. Methods in this investigation include basic physical properties such as enzyme digestibility, water absorption, water solubility, amylose-lipid complexing index, degree of substitution. In addition, differential scanning calorimetry (DSC), rapid viscosity analyzer (RVA), scanning electron microscope (SEM), texture profile analysis (TPA) were implemented for examining the gelatinization, retrogradation, gel structure, etc. Fourier transform infrared specscopy (FTIR) was further utilized for the quantitative characterization of ester bonds formed.
Resistivity to enzyme digestion and heat stability are two primary characteristics for the application of resistant starch. This study utilized hot water bath and autoclave to gelatinize mung bean starch, followed by retrograding the gelatinized starch at 4℃, forming stable double helical amylose and increased its crystallinity (RS3). Esterified starches (RS4) were
prepared by etherifying pregelatinized mung bean starch with low molecular weight carboxylic acids. It was anticipated to create the resistivity to enzyme digestion and heat stability through crosslinkage of ester bonds. The production of RS4 was found to exceed that of RS3 (retrograded starch), and the optimum condition was confirmed as reacted with sodium citrate. Further improvement on RS yield by the combination of physical and chemical treatments. Both water absorption index (WAI) and water solubility index (WSI) decreased upon storage which indicates the advance of retrogradation. WAI/WSI increased as the concentrations of citrate and reaction temperature climbed which were presumable attributed to the increase of hydrophilic ester bonds. Gelatinization of raw mung bean starch was found at 67℃ with 13.7 J/g enthalpy by differential scanning calorimeter (DSC). The peak in DSC thermogram disappears after gelatinization, yet moves to higher temperature zone with slight enhancement of transition enthalpy after retrogradation or esterification, which signifies the enhanced heat stability of resistant starch. The gelatinization temperature by RVA was close to that predicted by DSC, while setback viscosity of retrograded starch increased upon storage yet with a very low continuous viscosity, totally different from that of raw starch. Scanning electron microscope (SEM) confirmed the augmented network structure through gelatinization, but it became more compact through staling or esterification of starch. The network structure of starch gel almost fully disappeared by sodium citrate esterification. Absorption peak at 1728 cm-1 which could be attributed to ester bond was verified by Fourier transform infrared spectroscopy (FTIR), which proved that esterified starch acquires enzyme resistivity by its ester bonds.
In conclusion, RS3 through physical retrogradation, showed quite different characteristics in physicochemical aspects and structural evidences by spectroscopy, from that of RS4 which was chemically esterified. This may have an important effect on the application of these two RS.

中文摘要 I
Abstract III
謝誌 V
圖表目錄 X
1. 前言 1
2. 文獻回顧 2
2.1澱粉之介紹 2
2.1.1澱粉的組成 2
2.1.1.1直鏈澱粉 2
2.1.1.2支鏈澱粉 2
2.1.2澱粉之糊化 3
2.2抗性澱粉 9
2.2.1抗性澱粉的定義 9
2.2.2抗性澱粉之分類 9
2.2.3抗性澱粉的生成模式 10
2.2.4物理修飾產生的抗性澱粉 (澱粉之回凝) 11
2.2.4.1澱粉/水分含量 11
2.2.4.2加熱冷卻循環次數 15
2.2.4.3貯存時間與溫度 15
2.2.4.4 pH值 16
2.2.3.5食品成分 16
2.2.4.5.1油脂 16
2.2.4.5.2 鹽類 22
2.2.4.5.3 糖 22
2.2.4.6直鏈澱粉含量 22
2.2.4.7澱粉聚合度 24
2.2.4.7.1直鏈澱粉 24
2.2.4.7.2支鏈澱粉 24
2.2.5酵素處理產生抗性澱粉 24
2.2.5.1去分支酶 (Pullulanase) 25
2.2.5.2異澱粉酶作用方式 25
2.2.6化學修飾產生的抗性澱粉 25
2.2.6.1 RS4的介紹 25
2.2.6.2化學修飾澱粉 25
2.2.6.2.1無機酸水解修飾澱粉 25
2.2.6.2.2有機酸酯化澱粉 26
2.2.6.3酯化澱粉的使用規範 27
2.2.7抗性澱粉的研究近況 27
2.3 抗性澱粉在食品上的應用 33
2.3.1 在麵類食品中的應用 33
2.3.2 在焙烤食品中的應用 33
2.3.3 抗性澱粉在冷凍製品中的應用 34
2.3.4在飲料及發酵製品中的應用 34
2.4抗性澱粉在醫療上的應用 34
2.4.1 抗性澱粉與糖尿病 34
2.4.2 抗性澱粉與腸道疾病 35
2.4.3 抗性澱粉與體重控制 35
2.4.4 抗性澱粉與膽固醇、血脂質代謝 35
2.4.5 抗性澱粉與維生素、礦物質之吸收 35
2.5 綠豆 36
3.材料與方法 37
3.1實驗設計 37
3.2實驗材料 40
3.2.1原料 40
3.2.2試驗藥品 40
3.2.3試驗器材 41
3.3實驗方法 42
3.3.1實驗樣品的製備 42
3.3.1.1綠豆澱粉分離 42
3.3.1.2 RS3的製備 42
3.3.1.3 RS4 (酯化澱粉)的製備 42
3.3.1.3.2低分子羧酸鹽酯化澱粉的製備 42
3.3.1.3.3回凝酯化澱粉製備 43
3.4化學分析方法 43
3.4.1基本成分測定 43
3.4.1.1水分測定 43
3.4.1.2澱粉含量測定 43
3.4.1.3粗蛋白測定 43
3.4.1.4粗脂肪測定 43
3.4.1.5直鏈澱粉含量測定 44
3.4.1.6粗灰分測定 44
3.5抗性澱粉 (RS3、RS4)之物化特性 44
3.5.1抗性澱粉含量測定方法 44
3.5.2直鏈澱粉複合指數 (Amylose Complexing Index, ACI)測定 44
3.5.3澱粉之熱焓質(示差掃描熱分析, DSC) 47
3.5.4澱粉吸水性指標、水溶性指標之測定 47
3.5.5澱粉檸檬酸酯取代度的測定 48
3.5.6 FTIR傅立葉轉換紅外光譜 48
3.5.7連續糊化黏度之測定 48
3.5.8綠豆澱粉膠的物性分析 48
3.5.8.1質地特性測定 (Texture Profile Analysis, TPA) 48
3.5.8.2色澤分析 (L a b值的測定) 49
3.5.8.3掃描式電子顯微鏡觀察 (Scanning electron microscope, SEM) 49
3.5.9統計分析 49
4. 結果與討論 50
4.1綠豆澱粉基本成分分析 50
4.2抗性澱粉含量分析 50
4.2.1物理處理 50
4.2.2化學修飾 54
4.2.2.1不同低分子量羧酸鹽酯化澱粉 54
4.2.2.2檸檬酸鹽酯化澱粉 54
4.3吸水性指標與水溶性指標測定 57
4.3.1物理處理 57
4.3.2化學修飾 57
4.4示差掃描熱分析 (DSC) 63
4.4.1物理處理 63
4.4.2化學修飾 64
4.5連續糊化黏度測定 64
4.6直鏈澱粉複合指數 (amylose complexing index, ACI)測定 71
4.7檸檬酸酯取代度測定 71
4.8綠豆澱粉的物性分析 71
4.8.1掃描式電子顯微鏡 (SEM)觀察 74
4.8.1.1物理處理 74
4.8.1.2化學修飾 74
4.8.2質地剖面分析 (Texture Profile Analysis, TPA) 74
4.8.3色澤分析 (L a b值的測定) 82
4.8.3.1物理處理 82
4.8.3.2化學修飾 82
4.9酯化性質之相關性 83
4.10 FTIR官能基圖譜表現 83
5. 結論 89
參考文獻 91
作者簡介 102


圖表目錄
圖1、澱粉巨分子：（a）直鏈澱粉；（b）支鏈澱粉 5
圖2、澱粉顆粒內直鏈及支鏈澱粉分子分佈模式之一 6
圖3、連續黏度測定分析過程中澱粉顆粒變化之描述圖 7
圖4、連續糊化黏度分析儀典型曲線圖 8
圖5、抗性澱粉在直鏈澱粉溶液中，所生成的(a) 微胞體模與(b) 膠體模式 13
圖6、回凝澱粉構造模式圖 14
圖7、澱粉/水比例、加熱溫度及加熱冷卻循環次數對高直鏈玉米澱粉生產抗性澱粉含量之影響 17
圖8、以直鏈玉米澱粉經不同加熱/冷卻循環次數對抗性澱粉生成之影響 18
圖9、小麥澱粉經殺菌釜121℃加熱1小時後，於不同貯存溫度下其抗性澱粉含量之變化 19
圖10、未修飾與經檸檬酸修飾澱粉之傅立葉轉換紅外光譜圖 30
圖11、生澱粉、未修飾以及經戊二酸鈉修飾澱粉之傅立葉轉換紅外光譜圖 31
圖12、加熱前後天然玉米澱粉和經檸檬酸修飾玉米澱粉之掃描式電子顯微圖 32
圖13、不同回凝條件對抗性澱粉 (RS 3)生成與物化特性之影響 38
圖14、不同酯化條件對抗性澱粉含量 (RS 4) 生成與物化特性之影響 39
圖15、葡萄糖含量之標準曲線 45
圖16、直鏈澱粉含量之標準曲線 46
圖17、以不同糊化條件處理綠豆澱粉，於4℃下儲存對RS含量之影響 53
圖18、低分子量羧酸鹽和反應時間對綠豆澱粉生成抗性澱粉之影響 55
圖19、不同檸檬酸鹽濃度、反應溫度和反應時間對綠豆澱粉中抗性澱粉含量之影響 56
圖20、綠豆澱粉經水浴或高壓殺菌釜系統糊化後，於4℃貯存之吸水性指標 59
圖21、綠豆澱粉經水浴或高壓殺菌釜系統糊化後，於4℃貯存之水溶性指標 60
圖22、不同檸檬酸鹽濃度、反應溫度和反應時間對綠豆澱粉吸水性指標之影響 61
圖23、不同檸檬酸鹽濃度、反應溫度和反應時間對綠豆澱粉水溶性指標之影響 62
圖24、生綠豆澱粉之示差掃描熱分析圖譜 65
圖25、綠豆澱粉經沸水浴糊化或高壓殺菌釜系統糊化後於4℃貯存之示差掃描熱分析圖譜 66
圖26、綠豆澱粉經沸水浴糊化後，以檸檬酸鹽於不同溫度、不同時間進行酯化之示差掃描熱分析圖譜 68
圖27、不同檸檬酸鹽濃度、反應溫度和反應時間對綠豆澱粉中檸檬酸酯取代度之影響 73
圖28、綠豆澱粉經水浴糊化30 min後，於4℃貯存之電子顯微鏡照像圖 75
圖29、綠豆澱粉經水浴糊化60 min後，於4℃貯存之電子顯微鏡照像圖 76
圖30、綠豆澱粉經高壓殺菌釜糊化30 min後，於4℃貯存之電子顯微鏡照像圖 77
圖31、綠豆澱粉經高壓殺菌釜糊化60 min後，於4℃貯存之電子顯微鏡照像圖 78
圖32、不同濃度檸檬酸鹽、不同反應溫度反應3小時後酯化綠豆澱粉之掃描式電子顯微鏡圖 79
圖33、不同濃度檸檬酸鹽、不同反應溫度反應5小時後酯化綠豆澱粉之掃描式電子顯微鏡圖 80
圖34、0.2%檸檬酸鹽酯化澱粉之傅立葉轉換紅外光譜圖 87
圖35、0.4%檸檬酸鹽酯化澱粉之傅立葉轉換紅外光譜圖 88

表1、抗性澱粉之種類 12
表2、澱粉在營養上的分類 12
表3、香蕉澱粉經高壓蒸煮處理後，貯存於不同溫度和時間下之抗性澱粉含量、吸水性指標及水溶性指標 20
表4、澱粉/水懸浮液的pH值對抗性澱粉生成量之影響 21
表5、不同澱粉的直鏈澱粉含量和抗性澱粉生成量 23
表6、不同濃度、反應溫度、反應時間，對戊二酸鈉酯化薏仁澱粉的抗性澱粉生成量及顏色變化 28
表7、戊二酸鈉酯化薏仁澱粉加熱前後之抗性澱粉含量之變化 29
表8、綠豆澱粉之ㄧ般組成分析 51
表9、澱粉/水比例、兩種不同乾燥方法對綠豆澱粉中抗性澱粉產生量之影響 52
表10、綠豆澱粉經回凝酯化後對抗性澱粉含量生成之影響 58
表11、綠豆澱粉經不同糊化條件處理貯存於4℃不同時間之示差掃描熱分析特性 67
表12、檸檬酸酯化綠豆澱粉 (RS 4)在示差掃描熱分析中的特性關係 69
表13、綠豆澱粉經不同條件糊化後於4℃貯存不同時間之快速黏度測定儀測定之糊化特性 70
表14、綠豆澱粉經不同條件糊化後於4℃貯存不同時間之直鏈澱粉複合指數 72
表15、4℃冷藏對綠豆澱粉膠體膠強度的影響 81
表16、以不同糊化條件處理於4℃下儲存不同時間後，對綠豆澱粉色澤品質之影響 84
表17、不同酯化條件對綠豆澱粉色澤品質之影響 85
表18、RS含量、吸水性/水溶性指標、酯化取代度與色澤品質相關係數分析 86

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