臺灣博碩士論文加值系統

荔枝 (Litchi chinensis Sonn.) 屬無患子科 (Sapindaceae family)荔枝屬（Litchi）植物，為一種台灣重要的經濟作物，於三月下旬開花，六月下旬成熟。荔枝花一般都以廢棄物處理。我們先前的研究發現荔枝花的丙酮萃取物含有豐富的多酚類和極佳抗氧化力。因此本研究繼續將丙酮萃取物依次進行液液分離、矽膠管柱層析、LH-20管柱層析，所得的區分物均評估總酚與總類黃酮含量，及進行抗氧化活性測試 (總抗氧化力 (trolox equivalent antioxidant capacity, TEAC) 、DPPH (2,2-diphenyl-1-picrylhydrazyl hydrate) 自由基的清除能力和延遲Cu2+誘導LDL (low density lipoproteins) 的氧化能力) 。丙酮萃取物經液液萃取 (水、正丁醇、乙酸乙酯、正己烷) 以乙酸乙酯區分物的抗氧化活性較佳，且總酚及總類黃酮含量較高；再將此區分物經矽膠管柱層析得到13個區分，以第10~12區分物之抗氧化活性較好且總酚與總類黃酮含量較高；由第10~12區分再經 Sephadex LH-20管柱層各得到3個區分，以三者的第2區分最好；此三者之區分利用半製備型HPLC分離纯化得到兩個主要的抗氧化物質，經由Mass與NMR (nuclear magnetic resonance) 進行結構鑑定，確認荔枝花之主要抗氧化物質為表兒茶素(－)-epicatechin和原花青素proanthocyanidin A2。經由HPLC定量，凍乾的荔枝花 (－)-epicatechin 與proanthocyanidin A2含量分別為 5.48 及11.07 (mg/g of dry weight) 。

Litchi (Litchi chinensis Sonn.) belonging to Sapindaceae family is one of the important commercial crops in Taiwan, which blooms in late March and fruit matures in late June. The flower is usually considered as disposable byproducts. Our pervious study found that acetone extract of litchi flower had a notable amount of phenol and also exhibited good antioxidant capacity. In the investigation, the acetone extract was further separated through liquid-liquid partition, silica gel column chromatography and Sephadex LH-20 column chromatography in turn. The amounts of phenol and flavonoid in each collected fraction were determined; the antioxidant activities for each fraction were also assayed with Trolox equivalent antioxidant capacity (TEAC), scavenging ability on 2, 2-diphenyl-2-picrylhydrazyl hydrate (DPPH) radicals and inhibition of Cu2+-induced oxidation of human low density lipoprotein (LDL).
The acetone extract of litchi flower was separated by liquid-liquid partition into n-hexane, ethyl acetate, n-butanol, and water fractions. The ethyl acetate fraction had the highest phenol and flavonoid levels as well as antioxidant capacities. Thirty sub-fractions could be obtained after the ethyl acetate fraction was subjected to silica gel column chromatography. The sub-fractions 10~12 with higher phenol and flavonoid levels and antioxidant capacities were applied to Sephadex LH-20 column chromatography. Each sub-fraction (10, 11 and 12) could be further separated into three minor parts, and the minor-part 2 from each sub-fraction was the best, which could be isolated two major compounds by semi-preparative high performance liquid chromatography (HPLC). (-)-Epicatechin and proanthocyanidin A2 identified with Mass and nuclear magnetic resonance (NMR) were the major antioxidant compounds in litchi flower; their amounts were 5.48 and 11.07 (mg/g dry weight), respectively.

目錄
中文摘要.........................................................................................................................I
英文摘要.......................................................................................................................II
目錄..............................................................................................................................III
表次..............................................................................................................................IV
圖次............................................................................................................................IIV

壹、前言..........................................................................................................................1
貳、文獻整理.................................................................................................................3
一、荔枝與荔枝花之簡介.........................................................................................3
二、荔枝的生理功效與成分....................................................................................4
三、自由基與活性氧................................................................................................4
四、低密度脂蛋白與動脈粥狀硬化........................................................................5
五、抗氧化物質........................................................................................................6
六、酚類化合物之分離純化方法..........................................................................12
七、酚類化合物之抗氧化檢測..............................................................................15
参、研究目的與實驗架構..........................................................................................18
一、荔枝與荔枝花之簡介....................................................................................18
二、實驗架構........................................................................................................19
肆、材料與方法..........................................................................................................20
一、實驗材料..........................................................................................................20
二、實驗藥品..........................................................................................................21
三、實驗儀器..........................................................................................................23
四、實驗方法..........................................................................................................25
(一) 抗氧化含量分析...................................................................................25
1. 總酚類化合物 （Total polyphenols） 含量測定….................25
2. 總類黃酮 (Total flavonoids)含量測定.......................................25
(二) 抗氧化活性試驗..................................................................................26
1. DPPH自由基清除能力之測定.................................................................26
2. 總抗氧化能力（Trolox equivalent antioxidant capacity, TEAC）…….....26
3. 抑制銅離子誘導人類LDL氧化反應......................................................27
(三) 抗氧化成分之分離、純化..................................................................29
1. 溶劑萃取....................................................................................................29
2. 液液萃取....................................................................................................29
3. 矽膠管柱層析............................................................................................30
4.Sephadex LH-20管住層析…......................................................................30
5. 以高效液相層析儀 (HPLC) 分離純化荔枝花中主要抗氧化成分…...31
6. 荔枝花主要抗氧化成分含量之測定........................................................32
(四) 荔枝花主要抗氧化成分之結構鑑定.........................................................32
1. 紫外光-可見光光譜分析............................................................................32
2. 質譜檢測.....................................................................................................32
3. 1H-及13C-核磁共振光譜儀.........................................................................33
伍、統計分析............................................................................................................34
陸、結果與討論 ..........................................................................................................35
一、荔枝花丙酮萃取物進行液液萃取....................................................................35
二、 荔枝花丙酮萃取物之乙酸乙酯區分物進行矽膠管柱層析.........................40
三、Sephadex LH-20管柱層析...............................................................................46
四、HPLC分離、純化............................................................................................53
五、荔枝花主要抗氧化成分之結構鑑定................................................................55
六、荔枝花抗氧化物質含量分析............................................................................65
柒、結論........................................................................................................................67

Part 2
壹、文獻整理...............................................................................................................69
貳、研究目的與實驗架構...........................................................................................75
参、材料與方法............................................................................................................76
肆、結果與討論 ..........................................................................................................80
伍、結論........................................................................................................................82

參考文獻......................................................................................................................83
附錄..............................................................................................................................89

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