荔枝草之抗氧化能力研究 張逵揚、

荔枝草之抗氧化能力研究 張逵揚、

E-mail: [email protected]

摘 要

許多疾病被證實與體內氧化壓力（oxidative stress）有關，具有良好的抗氧化能力則可降低氧化壓力水平。荔枝草為唇型花 科二年生草本植物，在東亞地區如：中國、韓國、台灣、印度、馬來西亞與澳洲等皆能發現其分佈。在亞洲各國傳統民俗 用藥中，荔枝草被用來治療腫瘤、泌尿道感染、痔瘡，以及與發炎有關之疾病包含：肝炎、腹瀉及淋病。本研究使用四種 溶劑（水、甲醇、70%乙醇、乙酸乙酯）透過熱迴流萃取，將所得之萃取物進行抗氧化成分含量分析：總酚化合物與總類 黃酮，及抗氧化活性分析：DPPH自由基清除能力、ABTS陽離子自由基清除能力、超氧陰離子清除能力、亞鐵離子螯合能 力及還原力，評估荔枝草萃取物之抗氧化活性。細胞試驗則以細胞生存能力試驗來評估荔枝草之水萃物對Hep G2存活度的 影響。 實驗結果顯示，水萃物有最高萃取率，達16.3%；總酚化合物含量以70%乙醇萃取物為最高，達62.5±0.9 mg/g；總 類黃酮含量以乙酸乙酯萃取物為最高，達22.5±2.2 mg/g。於抗氧化活性方面，乙醇萃取物在DPPH自由基清除能力於濃 度0.1 mg/mL有最高清除活性（98.9%）；在抑制TBARS能力於濃度0.8 mg/mL有最高抑制率（86.1%）；在螯合能力於濃 度0.8 mg/mL有最高螯合率（69.5％）。荔枝草水萃取物在清除超氧陰離子能力於濃度1　mg/mL有到最高清除活性

（62.6%）。荔枝草甲醇萃取物在ABTS陽離子自由基清除能力於濃度0.5 mg/mL有最高清除活性，為99.9%；相對還原能 力於濃度0.8 mg/mL時與標準品BHA相比，有最高相對還原力（188.3％）。細胞生存能力試驗結果顯示，水萃物對Hep G2細胞的存活度沒有負面影響。綜合上述，荔枝草水、70％乙醇及甲醇萃取物具有良好的抗氧化能力，而水萃物對Hep G2沒有負面影響。

關鍵詞 : 荔枝草、氧化壓力、抗氧化能力、Hep G2

目錄

目錄 封面內頁 簽名頁 中文摘要iii 英文摘要v 誌謝vii 目錄viii 圖目錄xi 表目錄xii 1.緒論1 2.文獻回顧3 2.1氧化壓力3 2.2自由 基與活性氧4 2.2.1自由基4 2.2.2活性氧4 2.2.3氧化壓力指標5 2.3抗氧化防禦機制10 2.3.1一級防禦系統10 2.3.2二級防禦系 統10 2.4抗氧化劑之作用種類11 2.4.1自由基終止劑11 2.4.2還原劑或氧清除劑12 2.4.3金屬螯合劑12 2.4.4單重氧抑制劑13 2.5 荔枝草簡介13 2.5.1荔枝草外觀14 2.5.2荔枝草傳統藥效17 2.5.3荔枝草之研究17 2.6荔枝草主要成分19 2.7迷迭香酸21 3.材料 與方法24 3.1實驗材料24 3.2萃取物製備24 3.3試驗藥品25 3.4實驗設備27 3.5實驗架構28 3.6抗氧化成分含量分析30 3.6.1總酚 化合物30 3.6.2總類黃酮30 3.7抗氧化活性試驗31 3.7.1清除DPPH自由基能力31 3.7.2清除超氧陰離子能力32 3.7.3抑制硫代巴 比妥酸反應物生成（TBARS）能力32 3.7.4清除ABTS陽離子自由基能力33 3.7.5亞鐵離子螯合能力33 3.7.6還原能力34 3.8HPLC分析34 3.9細胞培養35 3.9.1培養基組成35 3.9.2細胞培養36 3.10細胞存活能力（Cell Viability）試驗37 3.11資料統計 分析37 4.結果與討論40 4.1荔枝草萃取率40 4.2總酚化合物與總類黃酮含量測定40 4.3抗氧化活性試驗44 4.3.1清除DPPH自 由基能力44 4.3.2清除超氧陰離子能力46 4.3.3清除ABTS陽離子自由基能力46 4.3.4抑制硫代巴比妥酸反應物生成能力49 4.3.5亞鐵離子螯合能力49 4.3.6還原力52 4.4HPLC成分分析54 4.5細胞存活能力試驗61 5.結論64 5.1結論64 5.2未來展望65 參 考文獻66 圖目錄 圖2.1氧分子的氧化還原及激發狀態8 圖2.2荔枝草標本15 圖2.3荔枝草全株16 圖2.4荔枝草主要類黃酮成 分20 圖2.5迷迭香酸結構式22 圖2.6迷迭香酸的合成路徑23 圖3.1實驗架構28 圖4.1荔枝草萃取物清除DPPH自由基之能力45 圖4.2荔枝草萃取物清除超氧陰離子之能力47 圖4.3荔枝草萃取物清除ABTS陽離子自由基之能力48 圖4.4荔枝草萃取物抑 制TBARS能力50 圖4.5荔枝草萃取物亞鐵離子螯合能力51 圖4.6荔枝草萃取物的相對還原能力53 圖4.7迷迭香酸HPLC之層 析圖55 圖4.8荔枝草水萃取物之HPLC層析圖56 圖4.9荔枝草乙醇萃取物之HPLC層析圖57 圖4.10荔枝草乙醇萃取物之HPLC 層析圖58 圖4.11荔枝草乙酸乙酯萃取物之HPLC層析圖59 圖4.12荔枝草萃取物對細胞存活度的影響（作用1小時）62 圖4.13 荔枝草萃取物對細胞存活度的影響（作用24小時）63 表目錄 表2.1活性氧物質之形7 表2.2生物體內氧化劑的種類9 表3.11×

磷酸鹽緩衝溶液配方38 表3.2Hep G2細胞培養基組成39 表4.1荔枝草不同溶劑之萃取率42 表4.2荔枝草不同萃取物之總酚與 總類黃酮含量43 表4.3HPLC分析荔枝草不同萃取物迷迭香酸含量60

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