臺灣博碩士論文加值系統

本研究利用澎湖地區所採集之腸石髮（Enteromorpha intestinalis）、海菜
（Monostorma nitidum）、石蓴（Ulva lactuca）、波狀網翼藻（Dictyopteris
undulate）、中國半葉馬尾藻（Sargassum hemiphyllum var. chinense）、凹頂藻
（Laurencia intermedia）及紫菜（Porphyra dentate）等七種海藻，使用熱水、
高溫高壓、均質及酵素等不同方法處理及萃取其機能性成份，並測定其總醣、總酚
含量，並利用DPPH(α,α-diphenyl-β-picryhydrazyl)自由基清除活性試驗、還原
力試驗、螯合亞鐵離子能力及SOD-like活性試驗等方法測定抗氧 化活性。
本研究結果顯示七種食用海藻含有許多總醣、總酚及高抗氧化活性物 質，因此，無
論熱水、殺菌及凍乾萃取，萃取物皆有明顯之抗氧化活性能力，特別是中國半葉馬尾
藻，無論在 DPPH 清除活性能力（76.6-98.6 %）、 還原力（1.9-2.9）及 SOD-like
(66.9-97.3 %)活性，比其他海藻萃取，皆顯示 較強之抗氧化能力。在海藻使用酵素
處理方面，海菜使用 Viscozyme 酵素 水解，其還原力2.88 為最高，而中國半葉馬尾
藻使用 Celluclast 醣解酶水解， 石蓴使用 Everlase 蛋白酶萃取，其 DPPH 自由基
清除能力各為92.2 %及98.8%，以上各組酵素萃取物皆高於對照組 Ascorbic acid、
Trolox 及其他海藻萃取，含有高抗氧化活性。此外，海藻以不同酵素萃取，則含有不
同性 質之抗氧化能力。利用海藻不同之抗氧化性質，開發海藻類保健產品，將有助於
推廣海 藻產品及增加海藻之利用價值。

In the present study, seven edible seaweeds including Enteromorpha
intestinalis, Monostorma nitidum, Ulva lactuca, Dictyopteris
undulate, Sargassum hemiphyllum var. chinense, Laurencia intermedia
and Porphyra dentate, were found to be rich in polysaccharide,
polyphenols, and also have high antioxidant activities.
Hot-water, sterilization, homogenization, lyophilization and enzyme
extraction, have significant effects on the antioxidant activity. The
antioxidant activity were analyzed by the methods such as DPPH scavening
activity, reducing power, chelating power, and SOD-like activity.
In this study, S. hemiphyllum var. chinense exhibited strong
antioxidant activity in DPPH (76.6-98.6%), reducing power (1.9-2.9), and
SOD-like activity (66.9-97.3 %). After enzyme digestion, M. nitidum treated
with Viscozyme was found to have the highest reducing power（2.88）.DPPH
scavening activities of the S. hemiphyllum var. chinense treated
with Celluclast carbohydrate hydrolyses and U. lactuca treated with
Everlase proteolytic hydrolyses, were 92.2 % and 98.8%,
respectively. Above mentioned all the enzyme
extracts, possessed higher antioxidant activities than those of the other
seaweeds extracts and the control.
Therefore, seaweed extracts with desired antioxidant properties,
could be used to develop seaweed products and increase value of seaweeds.

中文摘要---------------------------------------------------------i
英文摘要---------------------------------------------------------ii
致謝-------------------------------------------------------------iii
目錄-------------------------------------------------------------iv
表目錄-----------------------------------------------------------ix
圖目錄-----------------------------------------------------------x
第一章前言-------------------------------------------------------1
第二章文獻回顧---------------------------------------------------7
第一節
自由基與活性氧---------------------------------------------------7
一、自由基與活性氧之簡介-----------------------------------------7
1.超氧自由基（superoxide anion,˙O2•−）-------------------------9
2.過氧化氫（hydrogen peroxide, H2O2）--------------------------9
3.氫氧自由基(hydroxyl radical,˙OH)-----------------------------9
4.單重態氧（singlet oxygen , 1O2）-------------------------------10
5.過氧化自由基（peroxyl radical, ROO˙）-------------------------10
二、自由基與生物體之關係-----------------------------------------10
三、自由基與活性氧對生物體之氧化傷害-----------------------------11
四、自由基與疾病之相關性-----------------------------------------14
五、生物體之抗氧化防禦系統---------------------------------------14
1. 天然抗氧化物質-----------------------------------------------14
1.1 維生素C （ascorbic acid）----------------------------------14
1.2 維生素E （α-tocopherol）-----------------------------------15
1.3 β-胡蘿蔔素（β-carotene）-----------------------------------16
1.4 類黃酮（flavonoid）-----------------------------------------16
1.5 多酚類（polyphenols）---------------------------------------16
2. 抗氧化酵素---------------------------------------------------19
2.1 超氧歧化酶（superoxide dismutase, SOD）---------------------19
2.2 麩胱甘肽過氧化酶（glutathione peroxidase, GSH-Px）----------19
2.3 過氧化氫酶（catalase）--------------------------------------19
六、抗氧化劑之作用機制-------------------------------------------20
1.自由基終止型（free radical terminator）------------------------20
2.還原劑或氧清除劑者（reducing agent or oxygen scavenger）-------20
3.螯合劑型（chelating agent）------------------------------------21
七、酚類化合物的抗氧化效果及流行病學的關係-----------------------21
第二節
食用海藻介紹-----------------------------------------------------23
一、礁膜（Monostroma nitidum）-----------------------------------24
二、腸石髮（Ulva intestinalis Linnaeus）-------------------------25
三、石蓴（Ulva lactuca）-----------------------------------------26
四、波狀網翼藻（Dictyopteris undulata）--------------------------27
五、中國半葉馬尾藻(Sargassum hemiphyllum var. chinense)----------28
六、異枝軟骨凹頂藻（Chondrophycus intermedius）------------------29
七、長葉紫菜（Porphyra dentata）---------------------------------30
第三節
研究目的---------------------------------------------------------31
第三章 實驗材料與方法
一、藥品---------------------------------------------------------33
二、儀器設備-----------------------------------------------------33
三、材料---------------------------------------------------------34
四、海藻萃取-----------------------------------------------------34
1.熱水萃取-------------------------------------------------------34
2.殺菌-----------------------------------------------------------34
3.均質萃取-------------------------------------------------------34
4.冷凍乾燥萃取---------------------------------------------------35
5.酵素萃取-------------------------------------------------------35
五、體外抗氧化試驗-----------------------------------------------37
1.一般成分分析---------------------------------------------------37
1.1.水分---------------------------------------------------------37
1.2.灰分---------------------------------------------------------37
1.3.粗蛋白-------------------------------------------------------37
1.4.粗脂肪-------------------------------------------------------38
1.5.碳水化合物---------------------------------------------------38
2.海藻不同萃取物之總醣（total sugar）含量測定--------------------38
3.海藻不同萃取方法之總酚（total phenolic compond）含量測定-------39
4.海藻不同方法萃取物對清除α，α-diphenyl-β-picryl hydrazyl (DPPH)
自由基清除效力之測定-------------------------------------------40
5.海藻不同方法萃取物之類超氧歧化酶（SOD-like）活性測定-----------41
6.海藻不同方法萃取物之還原力（reducing power）測定---------------42
7.海藻不同方法萃取物之螯合亞鐵離子（chelating power）能力測定---43
8.DPPH、SOD-like 及還原力測定之 IC50 值--------------------------43
六、統計分析-----------------------------------------------------45
第四章
結果與討論-------------------------------------------------------46
一、海藻粉末一般成分分析-----------------------------------------46
二、海藻不同方法萃取物之總醣類含量-------------------------------46
三、海藻不同方法萃取物之總多酚類化合物含量-----------------------47
四、海藻不同方法萃取物對DPPH自由基之清除效力---------------------47
五、海藻不同方法萃取物之還原力-----------------------------------48
六、海藻不同方法萃取物之螯合亞鐵離子能力-------------------------49
七、海藻不同方法萃取物對類超氧歧化酶（SOD-like）活性-------------49
八、食用海藻酵素水解萃取之抗氧化活性、總醣及總酚含量-------------51
（一）醣解酶水解萃取---------------------------------------------51
1、DPPH自由基清除能力--------------------------------------------51
2、還原力--------------------------------------------------------51
3、螯合亞鐵離子能力----------------------------------------------51
4、SOD-like活性--------------------------------------------------52
5、總醣及總酚含量------------------------------------------------52
（二）蛋白酶水解萃取---------------------------------------------52
1、DPPH自由基清除能力--------------------------------------------52
2、還原力--------------------------------------------------------53
3、螯合亞鐵離子能力----------------------------------------------53
4、SOD-like活性--------------------------------------------------53
5、總醣及總酚含量------------------------------------------------53
第五章
結論-------------------------------------------------------------82
第六章
參考文獻---------------------------------------------------------83

Albert L. C., Chang C. K., Wu M. L., Huang. T. C. 2007. Studies on the
expression of liver detoxifying enzymes in rats fed seaweed (Monostroma
nitidum) Food Chem. Toxicol. 45, 2390-2396.
Amin Ismail, Zamaliah M. Marjan, Chin W. Foong. 2004. Total antioxidant
activity and phenolic content in selected vegetables. Food Chem. 87,
581-586.

Balasundram N., Sundram K., Samman S. 2006. Phenolic compounds in plants
and agri-industrial by-products: Antioxidant activity, occurrence, and
potential uses. Food Chem. 99, 191-203.

Betteridge D. John. 2000. What is oxidative stress. Metabol. Vol.49, No.2,
Suppl 1, pp. 3-8.
Cahyana A.H., Shuto Y., Kinoshita Y., 1992. Pyropheophytin a as an
antioxidative substance from the marine alga, Arame (Eisenia bicyclis).
Biosci. Biotechnol. Biochem. 56, 1533-1535.
Chandini S. Kumar, Ganesan P., Bhaskar N., In vitro antioxidant activities
of three selected brown seaweeds of India. 2008. Food Chem. 107, 707-713.
Chang C. H., Lin H. Y., Chang C. Y., Liu Y. C. 2006. Comparisons on the
antioxidant properties of fresh, freeze-dried and hot-air-dried tomatoes.
J. Food Engi. 77, 478-485.
Chaovanalikit A., Wrolstad R.E. 2004. Total anthocyanins and total phenolic
of fresh and processed cherries and their antioxidant properties. J. Food
Sci. 69, pFCT 67-72.
Christine D., Rainer S., Gerhard J. 2007. Amino acids, fatty acids, and
dietary fibre in edible seaweed products. Food Chem. 103, 891-899.
Chua Me. T., Tung Y. T., Chang S. T. 2008. Antioxidant activities of
ethanolic extracts from the twigs of Cinnamomum osmophloeum Bioresource
Technol.
99, 1918-1925.

Coothan K. V. , Anthony J., Sreenivasan P. P., Palaninathan V. 2007.
Benefcial role of sulfated polysaccharides from edible seaweed Fucus
vesiculosus in experimental hyperoxaluria. Food Chem. 100, 1552-559.
Decker E. A. and Welch B. (1990) Role of ferritin as a lipid oxidation
catalyst in muscle food. J. Agric. Food Chem. 38: 674-677.
Dorman H. J. D., Peltoketo A., Hiltunen R., Tikkanen M. J. 2003.

Characterisation of the antioxidant properties of de-odourised aqueous
extracts from selected Lamiaceae herbs. Food Chem. 83, 255-262.
Dubois M, gillers KA, Revers Ap, Smith F. 1956. Calorimetric method for
determination of sugars and related substance. Anal. Chem. 28 (4) 350-356.
Fang Y. Z., Yang S., and Wu G., PhD. 2002. Free Radicals, Antioxidants, and

Nutrition. Nutr. 18, 872-879.

Fang Z., Hu Y., Liu D., Chen J., Ye X. 2008. Changes of phenolic acids and
antioxidant activities during potherb mustard (Brassica juncea, Coss.)
pickling. Food Chem. 108, 811-817.
Fuller C. J., Grundy S.M., Norkus E. P. and Jialal I. 1996. Effect of
ascorbate supplementation on low density lipoprotein oxidation in smokers.
Atherosclerosis 119, 139-150.
Ganesan P., Kumar Chandini S., Bhaskar N.. 2008. Antioxidant properties of
methanol extract and its solvent fractions obtained from selected Indian
red seaweeds. Bioresource Technol. 99, 2717-2723.
Gray J. J., Gomaa E. A., Buckley D. J. 1996. Oxidative quality and shelf
life of
meats. Meat Sci. 43, 111-123.

Halliwell B. 1994. Free radicals and antioxidants: a personal view. Nutr.
Rev.

52(8), 253-265.

Halliwell B. and Gutteridge J. M. C. 1989. Free radicals, aging and

disease. In “Free Radicals in Biology and Medicine”, B. Halliwell, and

J. M. C. Gutteridge, Eds., Clarendon Press, Oxford. pp. 484-487.

Halliwell B. and Gutteridge J. M. C. 1998. The chemistry of free radicals
and related reactive species. In “Free Radicals in Biology and Medicine.”
B. Halliwell and J. M. C. Gutteridge, Eds., Clarendon Press, Oxford. pp.
36-104.

Halliwell B., Murcia M. A., Chirico S. and Aruoma O. I. 1995. Free radicals
and antioxidants in food and in vivo: what they do and how they work. Crit.
Rev. Food Sci. Nutr. 35: 7-20.
Heo Soo-Jin, Park Eun-Ju, Lee Ki-Wan, Jeon You-Jin. 2005. Antioxidant

activities of enzymatic extracts from brown seaweeds. Bioresource Technol.

96, 1613-1623.

Hirose M., Imaida K., Tamano S., and Ito N. (1994) Cancer chemoprevention
by antioxidants. In “ Food phytochmicals for cancer preventionⅡ”, C. T. Ho,

M. T. Huang and T. Osawa, Eds., Washington, DC: ACS. pp. 122-132. Jasna
Bertoncelj, Ursˇka Dobersˇek, Mojca Jamnik, Terezija Golob. 2007.
Evaluation of the phenolic content, antioxidant activity and colour of

Slovenian honey. Food Chem. 105, 822-828.

Javanmardi J., Stushno C., Locke E., Vivanco J.M. 2003. Antioxidant
activity and total phenolic content of Iranian Ocimum accessions. Food
Chem. 83,
547-550.
Katalinic V., Milos M., Modun D., Music I., M. Boban. 2004. Antioxidant
effectiveness of selected wines in comparison with (+)-catechin. Food Chem.
86, 593-600.

Keyvan D., H.J. Damien Dormana, Paivi P. Oinonen, Yusrida Darwis, Into
Laakso, Raimo Hiltunen. 2008. Chemical composition and in vitro
antioxidative activity of a lemon balm (Melissa officinalis L.) extract.
LWT
41, 391-400.

Kumar K. Suresh, Ganesan K., Rao P.V. Subba. Antioxidant potential of
solvent extracts of Kappaphycus alvarezii (Doty) Doty- An edible seaweed.
2008.
Food Chem. 107, 289-295.

Lahaye M., Kaffer B. 1997. Seaweed dietary fibres structure physiochemical

and biological properties relevant to intestinal physiology. Sci. Aliments.
17,

563-564.

Liu Q., Yao H. 2007. Antioxidant activities of barley seeds extracts. Food
Chem.

102, 732-737.

Lu M. J. a,b, Chen C. 2008. Enzymatic modi.cation by tannase increases the
antioxidant activity of green tea. Food Res. Int. 41, 130-137.
Mamath B.S., Namitha K.K., Senthil Amudha , Smitha J., Ravishankar G.A.

2007. Studies on use of enteromorpha in snack food. Food Chem. 101,

1707-1713.

Miller N. J., Evans C. A. R. 1996. The relative contributions of ascorbic
acid and phenolic antioxidants to the total antioxidant activity of orange
and apple fruit juices and blackcurrant drink. Food Chem. 60(3): 331-337.
Matsukawa R., Dubinsky Z., Kishimoto E., Masak K., Masuda Y., Takeuchi T.,
Chihara M., Yamamoto Y., Niki E., Karube I., 1997. A comparison of
screening methods for antioxidant activity in seaweeds. J. Appl. Phycol. 9,

29-35.

Meyer A.S., Heinonen M., Frankel E. N. 1998. Antioxidant interactions of
catechin, cyanidin, caffeic acid, quercetin, and ellagic acid on human LDL
oxidation. Food Chem. Vol. 61, No. 1/2, pp. 71-75.
Meyer A. S., Isaksen. A. 1995. Application of enzymes as food antioxidants.

Trends. Food Sci. Technol. 6, 300-304.

Mo’nica B. Pe’rez , Natalia L. Caldero’ n, Clara A. Croci. 2007.

Radiation-induced enhancement of antioxidant activity in extractsof
rosemary (Rosmarinus o.cinalis L.) Food Chem. 104, 585-592.
Montserrat Gudiel Urbano, Isabel Gon˜i. 2002a. Bioavailability of nutrients
in rats fed on edible seaweeds, Nori (Porphyra tenera) and Wakame (Undaria
pinnatifada), as a source of dietary fibre. Food Chem. 76, 281-286.
Montserrat Gudiel Urbano, Isabel Gon˜i. 2002b. Effect of edible seaweeds
(Undaria pinnatifida and Porphyra ternera) on the metabolic activities of
intestinal microflora in rats. Nutr. Res. 22, 323-331.
Moore R.E., 1978. Algal nonisoprenoids. In: Scheuer, P.J. (Ed.), Marine
Natural

Products, Chem. and Biol. Perspective, vol. 1. Academic Press, New York,
pp. 44-171.
Negro C., Tommasi L., Miceli A. 2003. Phenolic compounds and antioxidant
activity from red grape marc extracts. Bioresource Technol. 87, 41-44.
Nisizawa K., Noda H., Kikuchi R., & Watamaba T., 1987. The main seaweed
foods in Japan. Hydrobiol. 151/152, 5-29.
Oki T., Masuda M., Furuta S., Nishibia Y., Terahara N., Suda I., 2002.

Involvement of anthocyanins and other phenolic compounds in
radical-scavenging activity of purple-fleshed sweet potato cultivars. J.
Food

Chem. Toxicol. 67, 1752-1756.

Oyaizu M. 1988. Antioxidative activities of browning products of
glucosamine fractionated by organic solvent and thin-layer chromatography.
Nippon Shokuhin Kogyo Gakkaishi 35(11), 771-775.
Paula C., Martha T. Beconi, Cristian O’Flaherty. 2005. Relationship between
total superoxide dismutase activity with lipid peroxidation, dynamics and
morphological parameters in canine semen. Animal Reproduction Science 86,

163-173.

Qi H., Zhang Q., Zhao T., Hu R., ZhangK. and Lia Z. 2006. In vitro
antioxidant activity of acetylated and benzoylated derivatives of
polysaccharide extracted from Ulva pertusa (Chlorophyta). Bioor. & Medi.
Chem. Letters 16,
2441-2445.

Ramalakshmi K., Kubra I. Rahath, Rao L. Jagan Mohan. 2008. Antioxidant
potential of low-grade coffee beans. Food Rese. Int. 41, 96-103.
Santas J., Carbo R., Gordon M.H., Almajano M.P. 2008. Comparison of the
antioxidant activity of two Spanish onion varieties. Food Chem. 107,
1210-1216.

Sara M., Graham W. S., Michelle L. T. 2007. Comparison of nutritive
chemistry

of a range of temperate seaweeds. Food Chem. 100, 1331-1336.

Schechter S. R. 1996. Fighting free radicals with antioxidant. Health Foods

Business 6, 28-29.

Spranger I., Sun B., Mateus A. M., Vitor de Freitas, Jorge M.
Ricardo-da-Silva.

2008. Chemical characterization and antioxidant activities of oligomeric
and polymeric procyanidin fractions from grape seeds. Food Chem. 108,
519-532.

Stoilova I., Krastanov A., Stoyanova A., Denev P. , Gargova S. 2007.

Antioxidant activity of a ginger extract (Zingiber officinale). Food Chem.

102, 764-770.

Sua X., Duana J., Jianga Y., Shib J., Kakuda Y. 2006. Effects of soaking
conditions on the antioxidant potentials of oolong tea. J. Food Composition
and Analysis 19, 348-353.
Takashi K., Makiko T., Tomoko H., Yoko A. 2005. Antioxidant properties of
dried ‘kayamo-nori’, a brown alga Scytosiphon lomentaria (Scytosiphonales,
Phaeophyceae). Food Chem. 89, 617-622.
Takashi K., Taeko K., Hideyuki G., Takamoto S., Toshihiro Y. 2007.
Varieties

of antioxidant and antibacterial properties of Ecklonia stolonifera and
Ecklonia kurome products harvested and processed in the Noto peninsula,
Japan. Food Chem. 103, 900-905.
Takeshi N., Takakiyo Y. 2003. Preparation and functional properties of
beverages made from sea algae. Food Chem. 81, 327-332.
Wang K. H., Lin R. D., Hsu F. L., Huang Y. H., Chang H. C., Huang C. Y.,
Lee

M. H. 2006. Cosmetic applications of selected traditional Chinese herbal
medicines. J. Ethnopharmacol 106, 353-359.
Willians W. B., Cuvelier M. E. and Berset C. (1995) Use of free radical
method to evaluate antioxidant activity. Technol. 28, 25-30.
Wondimu T.e, Ueno A., Kanamaru I., Yamaguchi Y., McCrindle R., Hanaoka K.

I. 2007. Temperature-dependent extraction of trace elements in edible brown
alga hijiki, Hizikia fusiforme. Food Chem. 104, 542-550.
Yamaguchi T., Takamura H., Matoba T. and Terao J. (1998) HPLC method for
evaluation of the free radical-scavenging activity of foods by using 1,

1-diphenol-2-picrylhydrazyl. Biosci. Biotecnol. Biochem. 62(6), 1201-1204.
Yen F. L., Wu T. H., Lin L. T., Cham T. M., Lin C. C. 2008. Concordance
between antioxidant activities and .avonol contents in different extracts
and fractions of Cuscuta chinensis. Food Chem. 108, 455-462.
Yuan Y. V., Walsh N. A. 2006. Antioxidant and antiproliferative activities
of

Extracts from a variety of edible seaweeds. Food and Chem. Toxicol. 44,

1144-1150.

Yu L., Zhao M., Wang J. s., Cui C., Yang B., Jiang Y., Zhao Q. 2008.

Antioxidant, immunomodulatory and anti-breast cancer activities of phenolic
extract from pine (Pinus massoniana Lamb) bark. Innovative Food Sci. Emer.
Technol. 9, 122-128.
Yu P., Li N., Liu X., Zhou G., Zhang Q., Li P. 2003. Antioxidant activity
and antihyperlipidemic effects of polysaccharide extracted from Ulva
pertusa (Chlorophyta). Pharmacol. Res. 48, 543-549.
Zhu W., Chiu L.C.M., Ooi V.E.C., Chan P.K.S., Ang P.O., Jr. 2004. Antiviral
property and mode of action of a sulphated polysaccharide from Sargassum
patens against herpes simplex virus type 2. Int. J. Antimicrobial Agents
24,
81-85.

利文耀，「發酵蔬果飲品之製備及其機能性評估」，大葉大學生物產業科 技學系，碩士論文，民國96年。
吳玉菁，「聚球藻抗氧化活性探討」，屏東科技大學食品科學研究所，碩士 論文，民國92年。
孫財前，「綠藻水萃物之抗氧化性與關鍵組成份」，静宜大學食品營養學
系，碩士論文，民國95年。

陳淑芳，「天然食品抗氧化劑之應用現況」，食品工業發展研究所電子報， 民國96年。
陳淑茹，「石蓮萃取物之抗氧化活性及抗致突變性研究」，静宜大學食品 營養學系，碩士論文，民國92年。
黃淑芳，台灣東北角海藻圖錄，國立台灣博物館，台北，民國八十九年。


潘書毓，「台灣產山藥之酚類含量及抗氧化特性與加工的影響」，中國文

化大學生活應用科學研究所，碩士論文，民國九十二年。 鄭富元，柳育澤，萬添春，陳志銘，林亮全，。酵素水解雞腿骨蛋白製備

活性胜肽之研究Ⅱ：抗氧化分析。台灣農業化學與食品科學，第四十 五卷第二期，84-90頁，96年4月。
賴雅萍，「複方中草藥茶之抗氧化及抗致突變性」，國立嘉義大學食品科 學研究所，碩士論文，民國93年。
謝昆霖，「番木瓜果實水萃取物之抗氧化能力研究」，静宜大學食品營養 學系，碩士論文，民國96年。
蔣宗哲，李桂圓，莊培榳，陳盈年，曾慶瀛，桑椹酒陳化期間抗氧化活性 之探討。台灣農業化學與食品科學，第四十四卷第五期，315-325頁，
95年10月。