臺灣博碩士論文加值系統

洛神花英文名：Roselle (Hibiscus sabdariffa Linnaeus)，為一年生草本灌木，相關報告指出其具有抗氧化、抗醣化、降血壓、預防心血管等相關的能力。本研究以台灣洛神花為原料，以不同溫度 (30℃、80℃、90℃) 水及不同濃度 (50%、70%) 乙醇萃取，並進行機能性成分(總多酚、總類黃酮、總花青素、總多醣、有機酸) 含量測定及功能性 (抗氧化能力、抑制醣化能力、抑制細菌生長活性)等評估，最後篩選出最佳的50% 乙醇粗萃物，並進行Amberlite XAD-7管柱層析之分離純化，分別得到含0.2%、20% Delphinidin-3-O-sambubioside (醣苷鍵結飛燕草素) 的區分物，並探討其水解前後的抗氧化能力、抑制醣化能力及抑制細菌生長能力的差異性。研究結果顯示 (A)﹒抗氧化能力：(1).Trolox當量抗氧化能力：水解後的0.2%、20%-Delphinidin 區分物Trolox當量抗氧化能力為 (359.7、400.7uM)，具有較高的Trolox含量。2.清除DPPH自由基能力：水解前的0.2%-Delphinidin-3-O- sambubioside 區分物 (EC50：64.7 ug/g) 優於水解後的0.2%-Delphinidin區分物 (EC50：94.1 ug/g)。而在20% 飛燕草素區分物中，則以水解後 (EC50：138.4ug/g) 是優於水解前的 (EC50：242.1ug/g)。(B)﹒在抑制醣化能力上：未水解的0.2%、20% (20%-Delphinidin-3-O-sambubioside)的區分物，其抑制率為 (9.1-25.3%、10.2-53.0%) 優於水解後(0.0%、0.0%)。 (C)﹒在抑制細菌生長活性 (大腸桿菌、金黃色葡萄球菌)上：當在濃度 (0.5 mg/ml)時，顯示水解後的 (20%-Delphinidin) 之區分物具有最佳的抑菌能力，其抑菌能力可達 (100%、100%) 優於未水解 (20%-Delphinidin-3-O-sambubioside) 之區分物 (87.7%、50.1%)，其次為水解後的 (0.2%-Delphinidin)之區分物其抑菌能力為(98.4%、42.7%)，皆優於水解前的 (0.2%-Delphinidin-3-O- sambubioside) 之區分物 (16.0%、39.4%)。研究結果以水解後的 (20%-Delphinidin)之區分物，其在清除ABTS自由基及抑菌能力上為最佳。但未水解的(20%-Delphinidin-3-O-sambubioside) 之區分物則在抑制醣化的能力上為最佳。顯示水解前後的(20%-Delphinidin-3-O-sambubioside、20%-Delphinidin) 之區分物皆具有其一定的清除ABTS自由基、抑制醣化及抑制細菌的抑制能力。最後，證明提高洛神花酒萃物中花青素含量更能增加其功能性，是值得且具有作為開發保健食品的潛力。

The roselle (Hibiscus sabdariffa L.) is a annual shrub, related report showed that it has antioxidant, anti-glycation, anti-hyper- tension and prevent cardiovascular diseases. This study of raw material is Taiwan’s roselle, which is extracted useing different solvent as (30℃, 80℃, 90℃) water and (50%, 70%) ethanol, and is to evaluate and analyze its functional components content (polyphenols, total flavonoids, total anthocyanine, polysaccharides, organic acid) and (antioxidant, anti-glycation and inhibit bacterial growth capacity). Finally, we select the best of 50% ethanol of crude extracts and is separated and purified by the XAD-7 column chromatography. Respectively, obtained the (0.2%, 20%) Delphini- din-3-O-sambubioside of fractionates, and to assess the differences of antioxidant capacity, anti-glycation, inhibit bacterial growth capacity of unhydrolyzed and hydrolyzed fractionates. The study showed (A) the antioxidant capacity : (1) Trolox equivalent antioxidant capacity : the hydrolyzed fractionates (0.2%, 20%-Delphinidin) ( 359.7, 400.7uM) are the best than unhydrolyzed fractionates (313.3, 400.7 uM)(2)DPPH free radical scavenging capacity: unhydrolyzed fractionate (0.2%-Delphinidin-3-o-sambubioside) (EC50：64.7 ug/g) is the best than hydrolyzed fractionate (EC50：94.1g/g), but the hydrolyzed fractionate (20%-Delphinidin) (EC50：138.4ug/g) is the best than the unhydrolyzed fractionate (EC50：242.1 ug/g). In addition, (B). anti-glycation capacity: unhydrolyzed fractionates (0.2%, 20%-Delphinidin-3-O-sambubioside) are the best (9.1-25.3%, 10.2-53.0%) than the hydrolyzed fractionates (0.0%, 0.0%). (C). Inhibition of bacterial growth capacity (Escherichia coli, Staphylococcus aureus) : In 0.5mg/ml of sample, the hydrolyzed fractionate (20%-Delphinidin) (100%、100%) is the best than the unhydrolyzed fractionate 20%-Delphinidin-3-O-sambubioside (87.7、 50.1%). the hydrolyzed fractionate (0.2%-Delphinidin) (98.4%、42.7%) is the best than the unhydrolyzed fractionate (16.0%、39.4%).The results studied that the hydrolyzed fractionate (20%- Delphinidin) is the best in antioxidant and inhibit bacterial growth capacity.But unhydrolyzed fractionate (20%-Delphinidin-3-O-sam- bubioside) is the best in anti-glycation capacity, it was showed the unhydrolyzed (20%-Delphinidin-3-O-sambubioside) and hydrolyzed (20%-Delphinidin) fractionates are the better in antioxidant, anti-glycation and inhibit bacterial growth capacity. Finally, it is proven to the higher content of anthocyanin of ethanol extract that can be increased the Roselle’s functional, and have a potential as the development of health food products.

摘要..................................................................................................... Ⅰ
英文摘要.................................................................................................. Ⅲ
目錄..................................................................................................... Ⅴ
表目錄.................................................................................................. IX
圖目錄.................................................................................................. XI
壹、前言...................................................................................................1
貳、文獻回顧...............................................................................................4
一、洛神花的簡介與形態.......................................................................................4
二、洛神花的功能............................................................................................5
三、生理活性成分............................................................................................9
四、抑菌機制...............................................................................................15
五、花青素的純化方法........................................................................................17
參、研究目的與大綱..........................................................................................19
一、研究目的...............................................................................................19
二、實驗架構...............................................................................................20
肆、材料與方法.............................................................................................21
一、材料..................................................................................................21
(一) 原料...............................................................................................21
(二) 菌種...............................................................................................22
(三) 化學藥劑............................................................................................23
二、實驗儀器...............................................................................................24
三、方法...................................................................................................26
(一) 抗氧化能力...........................................................................................26
(二) 抑制醣化能力.........................................................................................28
(三) 抑制細菌生長試驗......................................................................................28
(四) 機能性成分分析........................................................................................30
(五) 50% 酒精洛神花萃取物之離子交換管柱層析分離純化方法.........................................................33
(六) 區分物水解...........................................................................................33
(八) 水解區分物測定其抑菌生長活性、清除DPPH自由基能力、Trolox抗氧化能力、抑制醣化能力...............................35
(九) 統計分析.............................................................................................35
伍、結果與討論...............................................................................................36
一、洛神花萃取率.............................................................................................36
二、洛神花各萃取物之功能性評估.................................................................................38
(一) 抗氧化能力...........................................................................................38
(二) 抑制醣化能力.........................................................................................45
(三) 抑制細菌生長試驗......................................................................................48
三、機能性成分分析...........................................................................................51
(一) 總多酚含量...........................................................................................51
(二) 總類黃酮含量測定......................................................................................51
(三) 總花素含量...........................................................................................52
(四) 總有機酸含量.........................................................................................52
(五) 總多醣含量...........................................................................................53
(六) 洛神花不同條件的萃取物所含機能性成分與功能性相關性分析......................................................55
(七) 篩選洛神花不同條件的萃取物之功能性和機能性最佳粗萃物........................................................59
四、洛神花之分離純化............................................................................................60
(一) 洛神花粗萃物之分離純化....................................................................................60
(二) 50% 乙醇粗萃物經管柱層析分離純化後之回收率..................................................................62
(三) 以HPLC 分析洛神花粗萃物及其區分物中含醣苷鍵結飛燕草素含量.....................................................64
(四) 以HPLC分析洛神花50% 乙醇粗萃物水解前後中含飛燕草素之區分物....................................................65
五、管柱分離純化含0.2%﹐20% 飛燕草素的洛神花區分物抑制細菌活性能力分析.................................................71
(一) 洛神花50% 乙醇粗萃物、含0.2%﹐20% 醣苷鍵結飛燕草素洛神花區分物抑制大腸桿菌活性試驗................................71
(二) 洛神花50% 乙醇粗萃物及0.2%、20% 不含醣苷鍵結區分物的水解物抑制大腸桿菌活性試驗...................................71
(三) 未水解的洛神花50% 乙醇粗萃物、0.2%、20% 含醣苷鍵結飛燕草素區分物抑制金黃色葡萄球菌活性之試驗.......................76
六、管柱分離純化含0.2%﹐20% 飛燕草素的洛神花區分物Trolox當量抗氧化能力................................................81
(一) 水解前後洛神花50% 乙醇粗萃物、0.2%、20% 的洛神花飛燕草素區分物其Trolox當量抗氧化能力的測定........................81
七、管柱分離純化含0.2%﹐20% 飛燕草素的洛神花區分物清除DPPH自由基.....................................................84
(一) 洛神花50% 乙醇粗萃物及0.2%、20% 洛神花飛燕草素的區分物的水解前後其清除DPPH自由基的EC50...........................84
八、管柱分離純化含0.2%﹐20% 飛燕草素的洛神花區分物抑制醣化能力........................................................86
(一) 洛神花50% 乙醇粗萃物及未水解的0.2%、20% 醣苷鍵結飛燕草素區分物其抑制醣化能力.....................................86
陸、結論......................................................................................................90
柒、論文出處...................................................................................................91

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