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研究生:陳偉齊
研究生(外文):Wei-Chi Chen
論文名稱:建立金針菇免疫調節蛋白 FIP-fve 品管指標及評估 FIP-fve 之生物可及性
論文名稱(外文):Establishment of Quality Control Indicator and Bioaccessibility Evaluation for Fungal Immunomodulatory Protein FIP-fve
指導教授:許輔許輔引用關係
口試委員:周志輝潘敏雄繆希椿
口試日期:2015-07-10
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:園藝學研究所
學門:農業科學學門
學類:園藝學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
語文別:中文
論文頁數:83
中文關鍵詞:FIP-fve分析方法確效火菇毒素生物可及性
外文關鍵詞:FIP-fveanalytical method validationflammutoxinbioaccessibility
相關次數:
  • 被引用被引用:2
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金針菇含有免疫調節蛋白 FIP-fve,具有抗過敏、抗腫瘤、免疫調節等生物活性,因此金針菇具有開發為健康食品素材的潛力。健康食品的開發必須要提出功效成分的科學證據,並能管理品質,使其在食用上須安全無虞。目前 FIP-fve 沒有一套完善的分析方法,從產品開發到品管皆有困難。因此,本研究的目的為建立分析 FIP-fve 的品管指標,測定金針菇中 FIP-fve 的含量,作為產品開發和品管依據,同時評估金針菇產品在食用上的安全性,並研究 FIP-fve 的生物可及性。
第一部分,建立 HPLC 分析 FIP-fve 的方法,並予以確效,並評估專一性、線性、準確度、精密度和偵測極限。結果顯示此分析方法穩定且值得信賴,並應用於後續的分析。第二部分,研究乾燥金針菇中 FIP-fve 的含量變化,找出最適當的乾燥條件用於往後的大量生產,結果發現金針菇分別經真空冷凍乾燥、真空微波乾燥和熱風乾燥後,三者 FIP-fve 含量並無顯著差異,且在熱風乾燥時 FIP-fve 含量會隨著溫度上升而顯著的下降。第三部分,探討食用安全性,對滅菌處理及金針菇的火菇毒素 (flammutoxin, FTX) 進行研究。首先將金針菇產品以加馬輻射滅菌,發現並不影響 FIP-fve 的含量。另一方面,火菇毒素為金針菇具溶血活性的毒蛋白,實驗發現火菇毒素於冷凍乾燥的金針菇中含量最多,其次為熱風和微波乾燥,而當熱風乾燥溫度提高時火菇毒素含量亦會顯著的下降。利用小鼠紅血球測其溶血活性,發現在熱風乾燥 70℃ 後,仍具有溶血活性。為解決火菇毒素的問題,初步以 60℃ 熱水預處理 5 分鐘後再進行50℃ 熱風乾燥,可使火菇毒素含量顯著下降並保留大部分的 FIP-fve。第四部分,在個別模擬胃及腸道消化的模式,在胃蛋白酶和胰酶的作用下,FIP-fve 僅少部分被消化。使用連續式胃腸模擬消化 FIP-fve,其在胃及腸道各消化兩小時後,FIP-fve 並不會降解。綜合以上結果,本實驗對金針菇健康食品開發從品管、安全性到消化已做一全面性的研究,證明金針菇具有作為健康食品的可行性。


Enoki mushroom was known as containing FIP-fve, a fungal immunomodulatory protein with several biological activities including anti-allergy, anti-tumor, and immunomodulation, and it showed a high potential as a good functional ingredient. The development of a successful health food product demanded the necessary requirements: (1) its ingredients should provide healthy and functional benefits with significant science evidences, (2) its functional compartments should be determined, and (3) the levels of the bio-activating compartments should be assured. Nevertheless, it lacked an appropriate analytical method available for FIP-fve analysis currently, which limited the utilization and application of enoki mushroom as a functional ingredient. Therefore, the aims of this study were to establish and to validate an analytical method as a quality control indicator of FIP-fve determination. In addition, the processing technologies to dehydrate the mushroom, to retain most of FIP-fve contents, and to eliminate the activity of hemolytic protein flammutoxin (FTX) were researched. Moreover, the bioaccessibility of FIP-fve within enoki juice and products were performed in this study.
First, we established and validated a HPLC method for FIP-fve determination. Through considering its specificity, linearity, accuracy, precision and quantitation limit, the results showed that this method was reliable and could be applied for subsequent analysis. Second, we studied the variation of FIP-fve contents in dried enoki mushroom, to figure out the best dehydration conditions for industrial production. The contents of FIP-fve showed no significant difference among vacuum freeze drying, vacuum microwave drying and hot air dehydration methods. Moreover, content of FIP-fve would be markedly dropped while the dehydration temperature was risen. Third, we investigated the processing stability of FIP-fve and FTX. Mushroom products exposed to gamma radiation presented no discernible difference in FIP-fve content. In addition, freeze dried mushroom yielded the highest level of FTX, which was followed by hot air dehydration and microwave drying, respectively. As the same as FIP-fve, content of FTX was dropped significantly while temperature rising and it was found that the hot air dehydration at 70℃ could not completely destroy the hemolytic activity of FTX. In order to provide a safe mushroom preparation, the mushrooms were preheated and treated using hot water at 60℃ before hot air dehydration at 50℃. Under this hot-water treatment, content of FTX dropped significantly and retained most of the FIP-fve levels. Forth, FIP-fve was found to be slightly degraded when using the model of mimicked stages of digestion in stomach and intestine. FIP-fve was not degraded when using the mimicked continuous gastrointestinal digestion model. In conclusion, this study made comprehensive researches of FIP-fve preparation and provided more evidences for enoki mushroom as a functional food ingredient for health food utilization.


目錄
口試委員會審定書 I
誌謝 II
摘要 IV
Abstract VI
目錄 VIII
表目錄 X
圖目錄 XI
第一章 前言 1
第一節 金針菇介紹 1
第二節 金針菇免疫調節蛋白 FIP-fve 5
第三節 分析方法確效 10
第四節 生物可及性 12
第五節 研究動機與目的 13
第二章 材料與方法 15
第一節 金針菇免疫調節蛋白 FIP-fve 純化 17
第二節 SDS-PAGE 18
第三節 高效液相層析儀 (HPLC) 分析 FIP-fve 方法確效 19
第四節 金針菇分析前處理方法 22
第五節 金針菇乾燥方法 23
第六節 膠體內水解 24
第七節 火菇毒素溶血活性分析 25
第八節 體外模擬胃消化金針菇汁 26
第九節 體外模擬腸道消化金針菇汁 26
第十節 體外模擬連續胃腸道消化金針菇汁 27
第十一節 高效液相層析儀 (HPLC) 分析火菇毒素 28
第十二節 統計分析 29
第三章 結果 31
第一節 建立及確效 HPLC 分析 FIP-fve 方法 31
第二節 評估金針菇乾燥後 FIP-fve 的含量 33
第三節 金針菇素材食用安全性 34
第四節 金針菇汁中 FIP-fve 之生物可及性 37
第四章 討論 40
第一節 建立 FIP-fve 品管指標 40
第二節 金針菇乾燥加工 41
第三節 乾燥金針菇食用安全性 42
第四節 金針菇汁中 FIP-fve 之生物可及性 45
第五章 結論與未來展望 47
參考文獻 48
TABLES 57
FIGURES 64



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