臺灣博碩士論文加值系統

本研究目的在於探討以表層海水 (surface sea water, SSW) 與深層海水 (deep sea water, DSW) 培養海洋擬球藻，其萃取物的組成差異和抗疲勞效果。在相同培養條件下，以表層海水及深層海水培養的海洋擬球藻其生長速率及外觀差異不明顯，但一般成分分析顯示，深層海水組的粗蛋白和碳水化合物含量較高，而表層海水組的灰分和粗脂肪含量較高，顯示以不同海水培養海洋擬球藻對其生長是有影響的。經超音波破胞後的海洋擬球藻萃取物，以深層海水培養的組別其水溶性蛋白質及總醣含量高於以表層海水培養組，而灰分含量則以表層海水培養組較高。萃取物中鈉含量以深層海水培養組高於以表層海水培養組 (p&;lt;0.05)，而其他礦物質含量皆無顯著差異。抗氧化能力的部分，深層海水組皆高於表層海水組之海洋擬球藻 (p&;lt;0.05)，可能是萃取物中多糖的含量與組成和游離胺基酸組成不同所造成。抗疲勞動物實驗結果中，餵食深層海水培養之海洋擬球藻萃取物在乳酸升高率明顯低於空白組 (p&;lt;0.05)，然而在其他抗疲勞評估項目中，如運動力竭試驗、乳酸消除率、血尿素氮含量以及肝臟中肝醣含量，餵食海洋擬球藻萃取物與空白組間皆無顯著差異，因此整體來說，海洋擬球藻萃取物的抗疲勞作用上並不顯著，但以深層海水培養之海洋擬球藻萃取物是具有潛在性的抗疲勞效果。

The purpose of this study is to compare the proximate composition and anti-fatigue effect of the extracts of Nannochloropsis oculata cultured by surface sea water (SSW) and deep sea water (DSW). Under the same culture condition, the growth rate and the appearance of N. oculata cultured by different sea water showed insignificant difference. The crude protein and carbohydrate content of N. oculata cultured by DSW were higher than cultured by SSW, on the contrary, the crude fat and ash content higher in SSW cultured extracts than DSW cultured extracts. It showed N. oculata cultured by different sea water had influence on their growth. After the ultrasound treatment, the soluble protein and total sugar content of the extracts of N. oculata cultured by DSW were higher than SSW. However the ash content was vice versa for the different cultured extracts. Both extracts of N. oculata cultured by SSW and DSW did not showed any significant difference in the other mineral content, except for sodium content. The antioxidant capacity of DSW cultured N. oculata extracts were significantly higher than SSW cultured extracts. This may be causing with polysaccharide’s amount and composition, and composition of free amino acid of extract. The results of the animal experiments, showed the lactic acid increment of DSW group were significantly better than the control group. The exhausting time, lactic acid elimination, blood urea nitrogen and liver glycogen content were not significantly different from the treatment group and control group. The anti-fatigue effect of the extracts of N. oculata were not significant. However, in comparison to SSW, DSW had potential benefit for anti-fatigue effect.

摘要 I
Abstract II
目錄 III
表目錄 VII
附錄 VIII
1、前言 1
2、文獻回顧 2
2.1. 微藻 2
2.1.1. 海洋擬球藻簡介 2
2.1.2. 影響微藻的培養條件 2
2.1.3. 海洋擬球藻的應用 3
2.2. 深層海水 3
2.2.1. 深層海水的特性 4
2.2.2. 深層海水的應用 4
2.3. 抗疲勞作用 5
2.3.1. 疲勞定義 5
2.3.2. 疲勞指標 5
2.4. 自由基 6
2.5. 抗氧化能力與抗疲勞關係 7
2.6. 礦物質與抗疲勞關係 7
3、實驗材料 8
3.1. 材料來源 8
3.1.1. 藻種來源 8
3.1.2. 海水來源 8
3.1.3. 實驗動物來源 8
3.2. 實驗藥品 8
3.3. 實驗儀器設備 9
4、實驗方法 11
4.1. 海水中成分分析 12
4.1.1. 營養鹽含量 12
4.1.2. 主要礦物質含量 13
4.2. 海洋擬球藻之培養條件 14
4.3. 海洋擬球藻之生長狀況 14
4.3.1. 海洋擬球藻之生長曲線 14
4.3.2. 海洋擬球藻之比生長速率 14
4.4. 海洋擬球藻之水萃取方法 14
4.5. 成分分析 15
4.5.1. 水分含量 15
4.5.2. 灰分含量 15
4.5.3. 粗蛋白含量 15
4.5.4. 碳水化合物含量 15
4.5.5. 總醣含量 16
4.5.6. 水溶性蛋白質含量 16
4.5.7. 主要礦物質含量 16
4.5.8. 硫酸根含量 16
4.5.9. 總多酚含量 17
4.5.10. 游離胺基酸含量 17
4.6. 抗氧化能力測定 18
4.6.1. ABTS自由基清除能力 18
4.6.2. DPPH自由基清除能力 18
4.6.3. 螯合亞鐵離子能力 19
4.6.4. 還原力之測定 19
4.7. 抗疲勞動物實驗 20
4.7.1. 實驗小鼠 20
4.7.2. 實驗設計 20
4.7.3. 運動能力試驗 20
4.7.4. 生化試驗 21
4.7.5. 抗疲勞判定之基準 22
4.8. 統計分析 23
5、結果與討論 24
5.1. 以表層海水及深層海水培養海洋擬球藻 24
5.1.1. 不同海水中的營養鹽及礦物質含量 24
5.1.2. 以不同海水培養之海洋擬球藻其生長狀況 24
5.1.3. 不同海水培養海洋擬球藻的一般成分分析 25
5.2. 以超音波協同水萃取海洋擬球藻 25
5.2.1. 海洋擬球藻萃取物之成分分析 25
5.2.2. 海洋擬球藻萃取物之礦物質含量 26
5.2.3. 海洋擬球藻萃取物之游離胺基酸含量 26
5.2.4. 海洋擬球藻萃取物之抗氧化能力 26
5.2.5. 海洋擬球藻萃取物中之抗氧化成分 27
5.3. 海洋擬球藻萃取物之抗疲勞動物實驗 28
5.3.1. 小鼠體重變化 28
5.3.2. 運動能力試驗 28
5.3.3. 血乳酸試驗 29
5.3.4. 血尿素氮含量 29
5.3.5. 肝臟中肝醣含量 29
5.4. 海洋擬球藻之抗疲勞作用 30
6、結論 31
7、參考文獻 32
8、圖 38
9、表格 49
10、附錄 59

王龍，2005，運動性疲勞的消除，甘肅政法成人教育學院學報第二期，pp. 152-154。
行政院衛生署，2003，健康食品之抗疲勞功能評估方法，衛署食字第0920401629號，臺北市，臺灣。
吳一德，1999，高強度間歇運動補充不同飲料對血清生化值之影響，大漢學報第十三期，pp. 387-400。
吳思賢，2010，深層海水培養微藻製備生質柴油，國立臺灣海洋大學輪機工程學系碩士論文，基隆市，臺灣。
周慶功、劉偉，2007，力竭運動小鼠體內部分元素含量的變化，環境與健康雜誌第24卷第12期，pp. 997-998。
林正常，1995，從生理學談運動疲勞，中華體育季刊第九期，pp. 35-43。
林金榮、蔡惠萍，2007，深層海水在水產養殖之應用及發展潛力，農業生技產業季刊，pp. 60-63。
林葦鈴、吳東昇、劉俊彥，2008，不同水層對餌料性藻類生長之差異評估，水利產業研討會，pp. 73-78。
徐振豐，2008，藻到健康，文經出版社有限公司，p. 18，臺北市，臺灣。
袁景道，2001，海洋細菌Pseudomonas vesicularis MA103 之agarases發酵槽生產條件探討以及海藻多醣，國立臺灣海洋大學食品科學系碩士論文，基隆市，臺灣。
陳伸賢、洪銘堅、黃宏莆、林文勝，2005，台灣深層海水資源利用政策與產業推動政策，深層海水中日國際研討會，臺北市，臺灣。
陳宥睿，2007，市售抗疲勞飲料對耐久性運動之抗疲勞影響之研究，大仁科技大學環境管理研究所碩士論文，屏東縣，臺灣。
劉瑞祥，2006，在光生化反應器中以二階段策略培養微藻生產油脂之研究，國立成功大學化學工程學系碩士論文，臺南市，臺灣。
謝昆霖，2007，番木瓜果實水萃取物之抗氧化能力研究，靜宜大學食品營養系碩士論文，臺中市，臺灣。
羅愛雁，2008，二十一世紀藍金－深層海水產業，科技發展政策報導第二期，pp. 106-109。
行政院環境保護署環境檢驗所，2002，重金屬檢測方法總則，NIEA M103.01C，臺北市，臺灣。
行政院環境保護署環境檢驗所，2003，水中矽酸鹽檢測方法－鉬矽酸鹽比色法，NIEA W450.50B2，臺北市，臺灣。
行政院環境保護署環境檢驗所，2006，水中硝酸鹽氮檢測方法－分光光度計法，NIEA W419.51A，臺北市，臺灣。
行政院環境保護署環境檢驗所，2010，水中磷檢測方法－分光光度計／維生素丙法，NIEA W427.53B，臺北市，臺灣。
A.O.A.C. 2005. Official methods of analysis. 18th edition. Association of Official Analytical Chemists. Arlington, U.S.A.
Abe, T., Takiguchi, Y., Tamura, M., Shimura, J., Yamazaki, K.I., 1995. Effect of vespa amino acid mixture (VAAM) isolated from hornet larval saliva and modified VMM nutrients on endurance exercise in swimming mice improvement in performance and changes of blood lactate and glucose. Japanese Journal of Physical Fitness and Sports Medicine 44, 225-238.
Alwan, M.R., Krasilshchikov, O., Muzaffar, T., Shihabudin, T.M., 2011. Effects of selenium supplementation on the muscle contractile properties, fatigue and antioxidant enzymes activities of gastrocnemius muscles in trained and untrained rats. South Asian Journal of Experimental Biology 1, 226-232.
Andrés, M., Raúl, C., Luis, L., Mariane, L., 1992. Evaluation of marine microalga Nannochloropsis sp. as a potential dietary supplement. Chemical, nutritional and short term toxicological evaluation in rats. Nutrition Research 12, 1273-1284.
Atmaca, G., 2004. Antioxidant effects of sulfur-containing amino acids. Yonsei Medical Journal 45, 776-788.
Brummer, Y., Cui, S.W., 2006. Detection and determination of polysaccharides in foods, In: Stephen, A.M., Phillips, G.O., Williams, P.A. (Eds.) Food Polysaccharides and Their Applications. CRC Press Inc, Florida, U.S.A., p. 689.
Cadenas, E., Davies, K.J.A., 2000. Mitochondrial free radical generation, oxidative stress, and aging. Free Radical Biology and Medicine 29, 222-230.
Chattopadhyay, N., Ghosh, T., Sinha, S., Chattopadhyay, K., Karmakar, P., Ray, B., 2010. Polysaccharides from Turbinaria conoides: Structural features and antioxidant capacity. Food Chemistry 118, 823-829.
Dinis, T.C.P., Madeira, V.M.C., Almeida, L.M., 1994. Action of phenolic derivatives (acetaminophen, salicylate, and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. Archives of Biochemistry and Biophysics 315, 161-169.
Dodgson, K.S., 1961. Determination of inorganic sulphate in studies on the enzymic and non-enzymic hydrolysis of carbohydrate and other sulphate esters. The Biochemical journal 78, 312-319.
Eom, S.H., Kang, Y.M., Park, J.H., Yu, D.U., Jeong, E.T., Lee, M.S., Kim, Y.M., 2011. Enhancement of polyphenol content and antioxidant activity of brown alga eisenia bicyclis extract by microbial fermentation. Fisheries and Aquatic Sciences 14, 192-197.
Fan, L.D., Zhai, F., Shi, D.X., Qiao, X.F., Fu, X.L., Li, H.P., 2011. Evaluation of antioxidant properties and anti-fatigue effect of green tea polyphenols. Scientific Research and Essays 6, 2624-2629.
FDA, 2002. Draft guidance for industry and reviewers-Estimating the safe starting dose in clinical trials for therapeutics in adult healthy volunteers. U.S. Food and Drug Administration, Rockville, Maryland, U.S.A. pp. 6-7.
Fukami, K., Nishijima, T., Hata, Y., 1992. Availability of deep seawater and effects of bacteria isolated from deep sea water on the mass culture of food microalga Chaetoceros ceratosporum. Nippon Suisan Gakkaishi 58, 931-936.
Gobatto, C.A., de Mello, M.A.R., Sibuya, C.Y., de Azevedo, J.R.M., dos Santos, L.A., Kokubun, E., 2001. Maximal lactate steady state in rats submitted to swimming exercise. Comparative Biochemistry and Physiology-Part A: Molecular and Integrative Physiology 130, 21-27.
Goh, S.H., Yusoff, F.M., Loh, S.P., 2010. A comparison of the antioxidant properties and total phenolic content in a diatom, Chaetoceros sp. and a green microalga, Nannochloropsis sp. Journal of Agricultural Science 2, 123-130.
Gu, N., Lin, Q., Li, G., Qin, G., Lin, J., Huang, L., 2012. Effect of salinity change on biomass and biochemical composition of Nannochloropsis oculata. Journal of the World Aquaculture Society 43, 97-106.
He, N., Yang, X., Jiao, Y., Tian, L., Zhao, Y., 2012. Characterisation of antioxidant and antiproliferative acidic polysaccharides from Chinese wolfberry fruits. Food Chemistry 133, 978-989.
Jiao, G., Yu, G., Zhang, J., Ewart, H.S., 2011. Chemical structures and bioactivities of sulfated polysaccharides from marine algae. Marine drugs 9, 196-223.
Kafaie, S., 2012. Acute and sub-chronic toxicological assessment of Nannochloropsis oculata in rats. African Journal of Agricultural Reseearch 7, 1220-1225.
Kimura, M., Tai, H., Nakagawa, K., Yokoyama, Y., Ikegami, Y., Takeda, T. 2004. Effect of drinking water without salt made from deep sea water in lipid metabolism of rats. In Ocean'04 MTS/IEEE Technology Conference (Kobe, Japan), pp. 320-321.
Lee, S.P., Mar, G.Y., Ng, L.T., 2009. Effects of tocotrienol-rich fraction on exercise endurance capacity and oxidative stress in forced swimming rats. European Journal of Applied Physiology 107, 587-595.
Liu, Y.X., Guo, Y.M., Wang, Z., 2007. Effect of magnesium on reactive oxygen species production in the thigh muscles of broiler chickens. British Poultry Science 48, 84-89.
Meucci, E., Mele, M.C., 1997. Amino acids and plasma antioxidant capacity. Amino Acids 12, 373-377.
Murat, J.C., Serfaty, A., 1974. Simple enzymatic determination of polysaccharide (glycogen) content of animal tissues. Clinical Chemistry 20, 1576-1577.
Nordberg, J., Arnér, E.S.J., 2001. Reactive oxygen species, antioxidants, and the mammalian thioredoxin system. Free Radical Biology and Medicine 31, 1287-1312.
Ono, E., Cuello, J.L., 2007. Carbon dioxide mitigation using thermophilic cyanobacteria. Biosystems Engineering. 96, 129-134.
Oyaizu, M., 1986. Antioxidative activity of browning products of glucosamine fractionated by organic soluble and thin-layer chromatography. Nippon Shokuhin Kogyo Gakkaishi 35, 771-775.
Pal, D., Khozin-Goldberg, I., Cohen, Z., Boussiba, S., 2011. The effect of light, salinity, and nitrogen availability on lipid production by Nannochloropsis sp. Applied Microbiology and Biotechnology 90, 1429-1441.
Powers, S.K., Howley, E., 2007. Exercise Physiology - Theory and application to fitness and performance, 6th eds. McGraw-Hill Co. Inc., New York, U.S.A. pp. 408-420, 477-498.
Powers, S.K., Jackson, M.J., 2008. Exercise-induced oxidative stress: cellular mechanisms and impact on muscle force production. Physiological Reviews 88, 1243-1276.
Putman, C.T., Spriet, L.L., Hultman, E., Lindinger, M.I., Lands, L.C., McKelvie, R.S., Cederblad, G., Jones, N.L., Heigenhauser, G.J., 1993. Pyruvate dehydrogenase activity and acetyl group accumulation during exercise after different diets. American Journal of Physiology - Endocrinology and Metabolism 265, 752-760.
Qi, H., Zhao, T., Zhang, Q., Li, Z., Zhao, Z., Xing, R., 2005. Antioxidant activity of different molecular weight sulfated polysaccharides from Ulva pertusa Kjellm (Chlorophyta). Journal of Applied Phycology 17, 527-534.
Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., Rice-Evans, C., 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine 26, 1231-1237.
Rubner, M. (1883). Uber den Einfluss der körpergrösse auf stoff- und kraftwechsel. Zeitschrift für Biologie. 19, 535-562.
Shan, X.Z., Zhou, J.L., Ma, T., Chai, Q.X., 2011. Lycium barbarum polysaccharides reduce exercise-induced oxidative stress. International Journal of Molecular Sciences 12, 1081-1088.
Singleton, V.L., Orthofer, R., Lamuela-Raventós, R.M., 1999. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods in Enzymology 299, 152-178.
Speich, M., Pineau, A., Ballereau, F., 2001. Minerals, trace elements and related biological variables in athletes and during physical activity. Clinica Chimica Acta 312, 1-11.
Sun, C., Wang, J.-W., Fang, L., Gao, X.-D., Tan, R.-X., 2004. Free radical scavenging and antioxidant activities of EPS2, an exopolysaccharide produced by a marine filamentous fungus Keissleriella sp. YS 4108. Life sciences 75, 1063-1073.
Tsuchiya, Y., Watanabe, A., Fujisawa, N., Kaneko, T., Ishizu, T., Fujimoto, T., Nakamura, K., Yamamoto, M., 2004. Effects of desalted deep seawater on hematologic and blood chemical values in mice. The Tohoku Journal of Experimental Medicine 203, 175-182.
Vinatoru, M., 2001. An overview of the ultrasonically assisted extraction of bioactive principles from herbs. Ultrasonics Sonochemistry 8, 303-313.
Wagenmakers, A.J., Beckers, E.J., Brouns, F., Kuipers, H., Soeters, P.B., van der Vusse, G.J., Saris, W.H., 1991. Carbohydrate supplementation, glycogen depletion, and amino acid metabolism during exercise. American Journal of Physiology - Endocrinology and Metabolism 260, 883-890.
Wang, J., Li, S., Fan, Y., Chen, Y., Liu, D., Cheng, H., Gao, X., Zhou, Y., 2010. Anti-fatigue activity of the water-soluble polysaccharides isolated from Panax ginseng C. A. Meyer. Journal of Ethnopharmacology 130, 421-423.
Wang, J., Zhang, Q., Zhang, Z., Song, H., Li, P., 2010. Potential antioxidant and anticoagulant capacity of low molecular weight fucoidan fractions extracted from Laminaria japonica. International Journal of Biological Macromolecules 46, 6-12.
Wang, S.T., Hwang, D.F., Chen, R.H., Chen, Y.C., Liang, C.W., Lin, C.S., Tsai, M.L., 2009. Effect of deep sea water on the exercise induced fatigue of rats. Journal of Food and Drug Analysis 17, 133-141.
Werman, M.J., Sukenik, A., Mokady, S., 2003. Effects of the marine unicellular alga Nannochloropsis sp. to reduce the plasma and liver cholesterol levels in male rats fed on diets with cholesterol. Bioscience, Biotechnology, and Biochemistry 76, 2266-2268.
Williams, M., 2005. Dietary supplements and sports performance: Minerals. Journal of the International Society of Sports Nutrition 2, 1-7.
Yang, B., Zhao, M.M., Shi, J., Yang, N., Jiang, Y.M., 2008. Effect of ultrasonic treatment on the recovery and DPPH radical scavenging activity of polysaccharides from longan fruit pericarp. Food Chemistry 106, 685-690.
Yen, G.C., Chung, D.Y., 1999. Antioxidant effects of extracts from Cassia tora L. prepared under different degrees of roasting on the oxidative damage to biomolecules. Journal of Agricultural and Food Chemistry 47, 1326-1332.
Rajapakse, N., Mendis, E., Jung, W.K., Je, J.Y., Kim, S.K., 2005. Purification of a radical scavenging peptide from fermented mussel sauce and its antioxidant properties. Food Research International 38, 175-182.
Suetsuna, K., Ukeda, H., Ochi, H. 2000. Isolation and characterization of free radical scavenging activities peptides derived from casein. Journal of Nutritional Biochemistry 11, 128–131.