黃世佑臺灣大學：化學工程學研究所盧秀青Lu, Shing-quingShing-quingLu2007-11-262018-06-282007-11-262018-06-282005http://ntur.lib.ntu.edu.tw//handle/246246/52103本研究從新鮮桑椹果實及乾燥的洛神葵花萼粉末中萃取分離出花青素，並進行抗氧化活性測定。將樣品之甲醇粗萃取物經由快速液相層析儀(Flash chromatograph)作初步分離，以25%甲醇(含1%醋酸)及50%甲醇(含1%醋酸)溶離，再利用HPLC作定性定量分析，進一步以HPLC-PDA、LC/MS、NMR鑑定花青素之結構。鑑定結果顯示，桑椹中含有cyanidin 3-glucoside、cyanidin-3-rutinoside及pelargonidin 3- rutinoside，其中cyanidin 3-glucoside、cyanidin-3-rutinoside含量為237.0±14.26mg (100g FW)-1、158.8±8.80mg (100g FW)-1，因此每100克新鮮桑椹果實可分離出395.8±14.26mg，而乾燥洛神葵花萼粉末中，可得到花青素8.7±0.05mg/g Dry Wt.。抗氧化活性測定方面，以FRAP(Ferric Reducing Ability of Plasma)及抑制H2O2誘發老鼠腦漿脂質過氧化(Lipid peroxidation)等方法，評估甲醇粗萃取物及快速層析儀分離出的色層之抗氧化活性。在抑制脂質過氧化表現上，F1-25及F1-50的ID50(μg)為2.0、1.7，相較於Trolox的ID50 4.3μg，抑制效果相當顯著。This work focuses on the separation, purification and identification of anthocyanins, mulberry (Morus spp.) and roselle (Hibiscus Sabdariffa L.) grown in Taiwan. Their antioxidative activities were also determined and compared. Extracts from the fruit samples were fractionated by flash chromatography using 25 and 50% methanol (with 1% acetic acid each). Qualitatative and quantitative analysis of fractionated products were carried out by using HPLC and identified the structures by employing HPLC-PDA, LC-Mass as well as NMR. The results showed that the mulberry contained cyanidin 3-glucoside (237.0±14.26mg/100g FW), cyanidin 3-rutinoside (158.8±8.80mg/100g FW) and a trace amount of plargonidin 3-rutinoside. 8.7±0.05mg/g DW of anthocyanins were isolated form the dried roselle. The method of Ferric Reducing Ability of Plasma (FRAP) and the inhibition of H2O2-induced lipid peroxidation in the rat brain homogenates were employed to assess their antioxidant activities. The ID 50 values of F1-25 and F1-50 were 2.0 and 1.7 μg, respectively. Comparing to the Trolox (ID 50 4.3μg), the inhibition capability of lipid peroxidation of mulberry was remarkable誌謝…………………………………………………………………… Ⅰ 摘要……………………………………………………………………. Ⅲ Abstract………………………………………………………………… Ⅳ 目錄……………………………………………………………………. Ⅴ 圖索引………………………………………………………………… Ⅷ 表索引………………………………………………………………… Ⅹ 第一章 緒論…………………………………………………………… 1 第二章 文獻回顧……………………………………………………… 3 2.1桑椹簡介………………………………… …………………………3 2.2洛神葵之簡介……………………………………………………… 5 2.3花青素之簡介………………………………………. ……………...7 2.4花青素之結構…………… …………………………………………8 2.5影響花青素顏色與穩定度之因素……… ………………………13 2.6萃取與分析……………………… ………………………………..18 2.7花青素的代謝與分泌…………………………………… ………..19 2.8花青素的抗氧化活性……………………………………………..20 2.9花青素的生理活性………………………………………………..23 第三章 材料與方法 …………………………………………………26 3.1實驗流程……………………… …………………………………26 3.2實驗材料…………………………………………………. ……….26 3.3實驗藥品…………………………………………………. ……….26 3.4儀器裝置………………………………………………… ………..27 3.5消耗器材………………………………………………… ………..28 3.6實驗方法…………………………………………………. ……….29 3.6.1不同溶劑萃取新鮮桑椹果實的比較………………… ……….29 3.6.2還原能力的測定(FRAP)…………………………………. .…..30 3.6.3萃取………………………………………………………….. ...31 3.6.4快速層析…………………………………………………….. ...31 3.6.5高效液相層析HPLC (定性定量分析)…………………..……31 3.6.7 H2O2誘導老鼠腦部均質液脂質過氧化…………………..…..32 3.6.8快速層析儀裝置圖……………………………………………. 34 第四章 結果與討論…………… …………………………………… 35 4.1不同溶劑對新鮮桑椹果中花青素的萃取效果比較………… …..35 4.2快速層析……………………………………… …………………36 4.3以HPLC分析快速層析儀所分離出的色層…………… ………..40 4.4化合物的分析與鑑定……………………………………………. 42 4.5還原力的測定FRAP(Ferric Reducing Ability of Plasma)……… .54 4.6 H2O2誘導老鼠腦部均質液脂質過氧化………………………… 60 4.7定量分析………………………………………………………….. 64 第五章結論………………………………………… …………………..67 第六章參考文獻………………………………………… ……………681996610 bytesapplication/pdfen-US花青素桑椹洛神葵抗氧化活性快速層析anthocyaninmulberryroselleantioxidative activityflash chromatographythesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/52103/1/ntu-94-R92524020-1.pdf