臺灣博碩士論文加值系統

梅子，溫帶薔薇科果樹，是南投名產之ㄧ。到目前為止已經，蒐集八種梅子樣品，以梅子的不同部位區分為（1.梅花、2.梅皮、3.梅肉），以梅子的不同成熟度區分（4.青梅果、5黃梅果、6.熟梅果），以梅子不同的加工方式區分（7.青梅果醃漬品、8.青梅果殺菌萃出液）。梅子已經證實具有抗氧化的能力。然而，有關於梅子抗氧化能力的詳細機制這部份的資訊仍是缺乏的。
利用超微弱冷光分析儀，並藉由探針標的超微弱冷光方法可以偵測四種活性氧，分別是：超氧陰離子、羥自由基、脂質過氧化自由基和過氧化氫。這個系統可以評估清除超氧陰離子功效依序排列為梅皮(IC50 = 2.4μg) ＞ 梅肉(IC50 = 3.1μg) ＞ 黃梅果(IC50 = 3.3μg) ＞ 熟梅果(IC50 = 3.6μg) ＞ 青梅果醃漬品(IC50 = 3.8μg) ＞ 青梅果(IC50 = 4.9μg) ＞ 青梅果殺菌萃出液 (IC50 = 17μg) ＞ 梅花(IC50 = 29μg)；第二，評估清除羥自由基功效依序排列為梅花(IC50 = 3.6μg) ＞ 青梅果殺菌萃出液(IC50 = 14μg) ＞ 青梅果(IC50 = 15μg) ＞ 梅皮(IC50 = 16μg) ＞ 梅肉(IC50 = 17μg) ＞ 熟梅果(IC50 = 23μg) ＝ 黃梅果(IC50 = 23μg) ＞ 青梅果醃漬品(IC50 = 33μg)；第三，評估清除過氧化氫功效依序排列為黃梅果(IC50 = 260μg) ＞ 梅皮(IC50 = 290μg)＝ 熟梅果(IC50 = 290μg) ＞ 青梅果醃漬品 (IC50 = 305μg) ＞梅肉(IC50 = 310μg) ＞青梅果殺菌萃出液(IC50 = 620μg) ＞青梅果、梅花(沒效果)；最候，評估清除脂質過氧化物自由基功效依序排列為梅花(1.42 Trolox equivalent unit ;T.E.U ) ＞青梅果殺菌萃出液(0.36 T.E.U) ＞ 青梅果(0.19 T.E.U) ＞ 梅肉(0.18T.E.U) ＞梅皮(0.15T.E.U) ＞ 熟梅果(0.1T.E.U) ＞黃梅果(0.09T.E.U)＞青梅果醃漬品(0.08T.E.U)。總而言之，我們結果可以提供對這些梅產品抗氧化能力的了解，透過比較彼此之間自由基清除能力（RSA）。藉此得以提供大眾有利健康及選擇梅產品之重要資訊。

Mei fruit (Japanese apricot; Prunus mune Sieb. et Zucc.), a deciduous tree of genus Rosaceae , is one of famous special agricultural products in Nanto country. So far, a total of 8 samples have been isolated from the Mei fruit with different partition (1.Flower、2.Peel、3.Pulp) and different degrees of ripening (4.Green Mei、5. Ripe Mei、6.Full-ripe Mei ) and varying methods of ( 7.Green Mei Brine、8.Green antioxidative capacities. However, information pertaining to the mechanism of how this agent is involved for its antioxidative capacity is scanty.
With the availability of an ultraweak chemiluminescence （uwCL）analyzer, we report here a probe-based uwCL technique which is capable of detecting a panel of four ODFRs, namely: superoxide anion (O2-), hydroxyl radical (•OH), peroxyl radical (ROO•) and hydrogen peroxide (H2O2).This system could measure the scavenging efficiencies of O2- in a descending order of Peel (IC50 = 2.4μg)＞ Pulp(IC50 = 3.1μg)＞ Full-ripe Mei(IC50 = 3.3μg)＞ Ripe Mei(IC50 = 3.6μg)＞ Green Mei Brine(IC50 = 3.8μg)＞ Green Mei(IC50 = 4.9μg)＞ Green Mei-sterilized extract (IC50 = 17μg)＞ Flower(IC50 = 29μg)；Second, scavenging efficiencies of •OH in a descending order of Flower(IC50 = 3.6μg)＞ Green Mei-sterilized extract (IC50 = 14μg)＞ Green Mei(IC50 = 15μg)＞ Peel (IC50 = 16μg)＞ Pulp(IC50 = 17μg)＞ Ripe Mei(IC50 = 23μg)＝ Full-ripe Mei(IC50 = 23μg)＞ Green Mei Brine(IC50 = 33μg); Third, scavenging efficiencies of H2O2 in a descending order of Full-ripe Mei(IC50 = 260μg)＞ Peel(IC50 = 290μg)＝ Ripe Mei(IC50 = 290μg)＞ Green Mei Brine (IC50 = 305μg)＞Pulp(IC50 = 310μg)＞ Green Mei-sterilized extract (IC50 = 620μg)＞ Green Mei、Flower(no efficiency); Lastly, scavenging efficiencies of ROO• in a descending order of Flower(1.42 Trolox equivalent unit ;T.E.U )＞ Green Mei-sterilized extract (0.36 T.E.U)＞ Green Mei(0.19 T.E.U)＞ Pulp(0.18T.E.U)＞ Peel(0.15T.E.U)＞ Ripe Mei(0.1T.E.U)＞ Full-ripe Mei(0.09T.E.U)＞ Green Mei Brine (0.08T.E.U).
Taken together; our data provide insight into the antioxidative capacity of these Mei products to allowing actual comparison of the radical scavenging activities（RSA）between them. These data can yield valuable information to the public regarding the selection of Mei products that could benefit to their health.

目錄 （Contents）
指導教授推薦書 …………………………………………………
口試委員審定書 …………………………………………………
授權書 …………………………………………………………… iii
致謝 ……………………………………………………………… iv
縮寫表 …………………………………………………………… v
中文摘要 ………………………………………………………… vi
英文摘要 ………………………………………………………… vii
目錄 ……………………………………………………………… viii
第一章 簡介 …………………………………………………… 1
1.1 文獻回顧 …………………………………………… 3
1.1.1 梅子的成分 ……………………………… 3
1.1.2 梅子抗氧化或保健功效 ………………… 4
1.1.3 探針標的超微弱冷光分析法 …………… 6
1.2 研究目的 …………………………………………… 7
第二章 材料與方法 …………………………………………… 8
2.1 各類梅的製備 ……………………………………… 8
2.2 各種活性氧產生系統 ……………………………… 8
2.3 儀器 ………………………………………………… 9
2.4 分析原理 ………………………………………… 9
2.5 實驗步驟及試劑配製 …………………………… 11
第三章 結果 …………………………………………………… 18
3.1 以探針標的超微弱冷光方法所建立四
種以氧為中心的自由基的評統 ………………… 18
3.2 抑制曲線的繪製及IC50的計算 ………………… 21
3.3 以探針標的超微弱冷光分析法偵測各
類梅產品之自由基清除能力Radical
scavenging ability(RSA)之評估 ……………… 23
3.3.1 未加工梅子部分 …………………………… 24
3.3.2 加工梅子部分 ……………………………… 28
第四章 討論 …………………………………………………… 36
4.1 梅子不同部位與抗氧化的關係 …………………… 36
4.2 梅子不同成熟與抗氧化的關係 …………………… 37
4.3 梅子不同加工方式與抗氧化的關係 ……………… 38
第五章 結論 …………………………………………………… 39
第六章 參考文獻 ……………………………………………… 40

第六章 參考文獻 （References）
1.中華本草. 第四冊第十卷. 上海科學技術出版社 p.86-93, 1999.
2.張正明. 梅子醃漬前果汁之製取及脫除苦味之研究. 台大食科所論文, 1984.
3.方祖達、趙昭惠. 梅子果汁成分類型分布之研究. 食品科學21 （1）: 34-45, 1994.
4.沈紅梅、易楊華、喬傳卓、蘇中武、李承祜. 烏梅的化學成分研究. 中草葯 26（2）: 105-106, 1995.
5.沈紅梅、喬傳卓、蘇中武、李承祜. 採收、加工、植物基原對烏梅抑菌作用的影響. 中葯材 17（8）: 24, 1994.
6.沈紅梅、程濤、喬傳卓、蘇中武、李承祜. 烏梅的體外抗腫瘤活性及免疫調節作用初探. 中國中葯雜誌 20（6）: 365-368, 1995.
7.柏有成. 梅子核仁中β-glucosidase之純化與生化性質之探討. 台大園藝所碩士論文, 1989.
8.Hies H. Oxidative stress: oxidants and antioxidants. Exp Physiol. 82: 291-295, 1997.
9.Halliwell B. Free radical and antioxidants: A personal view. Nutri Rev. 52: 253-265, 1994.
10.Minotti G and Auat SD. The requirement for iron (III) in the initiation of lipid peroxide by iron (II) and hydrogen peroxide. J Biol Chem. 3: 1098-1104, 1987.
11.Perugini C, Seccia M, Alboano E. The dynamic reduction of Cu (II) to Cu (I) and not Cu (I) availability is a sufficient trigger for low density lipoprotein oxidation. Biochim Biophys Acta. 1343: 191-198, 1997.
12.Zarger RA and Burkhart RM. Differential effects of glutathione and cystein on Fe2+, Fe3+, H2O2 and myoglobin-induced proximal tubular cell attack. Kidney In+.53: 1661-1672, 1998.
13.Wanagat J, Cao Z, Pathare P, and Aiken JM. Mitochondrial DNA deletion mutations colocalize with segmental electron transport system abnormalities, muscle fiber atrophy, fiber splitting, and oxidative damage in sarcopenia. FASEB J. 15: 322–332, 2001.
14.Burdon R. Superoxide and hydrogen peroxide in relation to mammalian cell proliferation. Free Radical Biol Med. 18: 775–794, 1995.
15.Crawford DR, Zbinden I, Amstad P, and Cerutti P. Oxidant stress induces the proto-oncogenes c-fos and c-myc in mouse epidermal cells. Oncogene. 3: 27–32, 1988.
16.Datta R, Hallahan DE, Kharbanda SM, Rubin E, Sherman ML, Huberman E, Weichselbaum RR, and Kufe DW. Involvement of reactive oxygen intermediates in the induction of c-jun gene transcription by ionizing radiation. Biochemistry. 31: 8300–8306, 1992.
17.Nose K, Shibanuma M, Kikuchi K, Kageyama H, Sakiyama S, and Kuroki T. Transcriptional activation of early-response genes by hydrogen peroxide in a mouse osteoblastic cell line. Eur J Biochem. 201: 99–106, 1991.
18.Bae YS, Kang SW, Seo MS, Baines IC, Tekle E, Chock PB, and Rhee SG. Epidermal growth factor (EGF)-induced generation of hydrogen peroxide. J Biol Chem. 272: 217–221, 1997.
19.Ha HC, Thiagalingam A, Nelkin BD, and Caserora JR. Reactive oxygen species are critical for the growth and differentiation of medullary thyroid carcinoma cells. Clin Cancer Res. 6: 3783–3787, 2000.
20.Meier B, Radeke HH, Selle S, Younes M, Sies H, Resch K, and Habermrhl GG. Human fibroblasts release reactive oxygen species in response to interleukin-1 or tumor necrosis factor-a. Biochem J. 263: 539–545, 1989.
21.Suh YA, Arnold RS, Lassegue B, Shi J, Xu X, Sorescu D, Chung AB, Griendling KK, and Lambeth JD. Cell transformation by the superoxide- generating oxidase Mox1. Nature. 401: 79–82, 1999.
22.Sundaresan M, Zu-xi Y, Ferrans VJ, Irani K, and Finkel T. Requirement for generation of H2O2 for platelet-derived growth factor signal transduction. Science. 270: 296–299, 1995.
23.Szatrowski TP and Nathan CE. Production of large amounts of hydrogen peroxide by human tumor cells. Cancer Res. 51: 794–798, 1991.
24.Zafari AM, Ushio-Fukai M, Akers M, Yin Q, Shan A, Harrison DG, Taylor WR, and Griendling KK. Role of NADH/NADPH oxidasederived H2O2 in angiotensin II-induced vascular hypertrophy. Hypertension. 32: 488–495, 1998.
25.Harman D. Aging: a theory based on free radical and radiation chemistry. J Gerontol. 11: 298–300, 1956.
26.Brody JA. Prospects for an ageing population. Nature. 315: 463–466, 1985.
27.Campion EW. The oldest old. N Engl J Med 330: 1819–1820, 1994.
28.Lamberts SWJ, Van Den Beld AW, and VAN DER LELY AJ. The endocrinology of aging. Science. 278: 419–424, 1997.
29.Buscigllo J and Yankner BA. Apoptosis and increased generation of reactive oxygen species in Down’s syndrome neurons in vitro. Nature. 378: 776–779, 1995.
30.Buscigllo J and Yankner BA. Apoptosis and increased generation of reactive oxygen species in Down’s syndrome neurons in vitro. Nature. 378: 776–779, 1995.
31.Sinet PM, Coutrier J, and Dutillaux B. Trisomie 21 et superoxyde dismutase-1 (IPO-A): tentative de localisation sur la sous-bande21q22.1. Exp Cell Res. 97: 47–55, 1976.
32.Anneren G and Epstein CJ. Lipid peroxidation and superoxide dismutase-1 and glutathione peroxidase activities in trisomy 16 fetal mice and human trisomy 21 fibroblasts. Pediatr Res. 21: 88–92, 1987.
33.Brooksbank BWL and Balazs R. Superoxide dismutase, glutathione peroxidase and lipid peroxidation in Down’s syndrome fetal brain. Dev Brain Res. 16: 37–44, 1984.
34.De Haan JB, Christiano F, Iannello RC, and Kola I. Cu/Zn-superoxide dismutase and glutathione peroxidase during aging. Biochem Mol Biol Int. 35: 1281–1297, 1995.
35.De Haan JB, Wolvetang EJ, Christiano F, Iannello RC, Bladier C, Kelner MJ, and Kola I. Reactive oxygen species and their contribution to pathology in Down’s syndrome. Adv Pharmacol. 38: 379–402, 1997.
36.Flescher E, Ledbetter JA, Schieven GL, Vela-Roch N, Fossum D, Dang H, Ogawa N, and Talal N. Longitudinal exposure of human T lymphocytes to weak oxidative stress suppresses transmembrane and nuclear signal transduction. J Immunol. 153: 4880–4889, 1994.
37.Dogasaki C, Murakami H, Nishijima M, Yamamoto K, and Miyazaki T. Antimutagenic activities of hexane extracts of the fruit extract and the kernels of Prunus mume Sieb. et Zucc. Yakugaku Zasshi. 112:577-584, 1992.
38.Chuda Y, Ono H, Ohnishi-Kameyama M, Matsumoto K, Nagata T, and Kikuchi Y. Mumefural, citric acid derivative improving blood fluidity from fruit-juice concentrate of Japanese apricot (Prunus mume Sieb. et Zucc). J Agric Food Chem. 47: 828–831, 1999.
39.Utsunomiya H, Takekoshi S, Gato N, Utatsu H, Motley ED, Eguchi K, Fitzgerald TG, Mifune M, Frank GD, and Eguchi S. Fruit-juice concentrate of Asian plum inhibits growth signals of vascular smooth muscle cells induced by angiotensin II. Life Sci .
72: 659–667, 2002.
40.Matsuda H, Morikawa T, Ishiwada T, Managi H, Kagawa M, Higashi Y, and Yoshikawa M. Medicinal flowers. VIII. Radical scavenging constituents from the flowers of Prunus mume: structure of prunose III. Chem Pharm Bul . 51: 440–443, 2003.
41.Van Den Berg R, Haene GR, Van Den Berg H, and Bast A. Applicability of an improved Trolox equivalent antioxidant capacity (TEAC) assay for evaluation of antioxidant capacity measurements of mixtures. Food Chem. 66: 511-517, 1999.
42.Wayner DDM, Burton GW, Ingold KU, Barclay LRCI, and Locke JF. The relative contributions of vitamin E, urate, ascorbate and proteins to the total peroxyl radical-trapping antioxidant activity of human blood plasma. Biochim Biophy Acta. 924: 408-419, 1987.
43.Tsai CH, Chang RC, Chiou JF, and Liu TZ. Improved superoxide-generating system suitable for the assessment of the superoxide-scavenging ability of aqueous extracts of food constituents using ultraweak chemiluminescence. J Agric Food Chem. 51: 58-62, 2003.
44.Tsai CH, Stern A, Chiou JF, Chern CL, and Liu TZ. Rapid and specific detection of hydroxyl radical using an ultraweak chemiluminescence analyzer and a low-level chemiluminescence emitter: application to hydroxyl radical- scavenging ability of aqueous extracts of Food constituents. . J Agric Food Chem. 49: 2137-2141, 2001.