東京水産大学食品生産学科東京水産大学食品生産学科東京水産大学食品生産学科東京水産大学食品生産学

Takeshi Suzuki

Wei Wang

Ya-Pei Hsieh

Yumiko Yoshie

吉江 由美子

王 火韋

謝 雅珮

鈴木 健

Open Access Collection of International and Scholarly Papers

TUMSAT-OACIS Repository - Tokyo University of Marine Science and Technology (東京海洋大学)Compositional Difference of Phenolic Compoundsbetween Two Seaweeds, Halimeda sppjournal orpublication title東京水産大学研究報告volume 88page range 21-24year 2002-03-29URL http://id.nii.ac.jp/1342/00000100/Journal of Tokyo University of Fisheries, Vol. 88, pp. 21-24, 2002Compositional Difference of Phenolic Compounds between Two Seaweeds, Halimeda spp.Yumiko Yoshie,* Wei Wang,* Ya-Pei Hsieh* and Takeshi Suzuki* (Received August 29, 2001)Abstract: Halimeda macroloba and Halimeda opuntia were collected at their growing district (Ishigaki island, Oki-nawa Pref., Japan), in the same season. Polyphenolic and related phenolic compounds were extracted from two Hal-imeda, and analyzed by high performance liquid chromatography. Polyphenolic compounds (catechin, epicatechin,epigallocatechin, catechin gallate, epicatechin gallate, epigallocatechin gallate, rutin, quercitrin, hesperidin, myricetin,morin, luteolin, quercetin, apigenin, kaempferol, baicalein) and related phenolic compounds (caffeic acid and catechol)were determined. The composition of polyphenolic and related phenolic compounds was different between two Hal-imeda. The extremely large amount (28,000µg/g dry matter) of epigallocatechin was found in H. macroloba. Caffeicacid and hesperidin were found only in H. macroloba. Catechol was detected in H. macroloba 5 times as much as cate-chol in H. opuntia. Myricetin and morin were found in H. macroloba approximately double amounts from H. opuntia.Other polyphenolic compounds tested in this study were not found from both Halimeda. Keywords:  polyphenolic compounds, Halimeda, HPLC analysisIntroductionPolyphenolic compounds occur ubiquitously in foods ofplant origin. Since polyphenolic compounds have severalhydroxyl (OH) groups, it was expected to express radicalscavenging effect1,2) or sometimes to have prooxidanteffect as a source of reactive oxygen species. 3,4)Polyphenolic compounds may have beneficial healtheffects because of their antioxidant properties and theirinhibitory role in various stages of tumour development inanimal studies. There are many studies to reportpolyphenolic composition in teas5-8), wines8-10),cacaos11,12), fruits and vegetables13-17). There might besome high flavonoid/catechin concentration in seaweedswhich were not used for our life in comparison to the landplant. Previously we reported the contents of catechinsand their related compounds in seaweeds,18) there weretwo Caurelpa, but they did not have similarity in catechincomposition. In this report, there are two Halimeda; one isHiroha-sabotengusa (H. macroloba), the other isSabotengusa (H. opuntia) in Japanese. They are living inthe subtropical area, but are not used as a food likeCaurelpa. Halimeda is living in the seashore, and easy tocollect even by children. Therefore we tried to analyze thecontents for efficient utilization. In spite of the similarshape and living area each other, no one reported thedifferences of their contents. Thus, the purpose of thisresearch was to note the difference of polyphenoliccomposition in two Halimeda.Materials and MethodsSeaweed samplesGreen algae Halimeda macroloba and Halimedaopuntia were collected at their growing district, Ishigaki,Okinawa Prefecture, and transported under refrigeration.After washing with tap water and wiping with a papertowel, they were minced by a food cutter (MK-K75,Matsushita Electric Co., Osaka, Japan), and stored at -20°Cuntil use. ChemicalsAuthentic catechin, epicatechin, epigallocatechin,catechin gallate, epicatechin gallate, epigallocatechingallate, rutin, caffeic acid, catechol, hesperidin, quercitrin,myricetin, morin, luteolin, quercetin, apigenin,kaempferol, and baicalein were purchased from SigmaChemical Co. (St. Louis, MO, USA). All other chemicalswere commercially available.Extraction and Analysis of CatechinsCatechins were extracted according to the method fortea catechin7) as mentioned in the previous paper.18) Theextracts were kept at -80°C (freezer, Sanyo MDF-392)until analysis. All samples were extracted and analyzed intriplicate. Catechins were determined by a high-performanceliquid chromatograph modified from the methods ofSuematsu et al.6) and Terada et al.5) Catechins were* Department of Food Science and Technology, Tokyo University of Fisheries, 5-7, Konan 4-chome, Minato-ku, Tokyo 108-8477, JapanY. Yoshie, W. Wang, Y. Hsieh and T. Suzuki22separated by an ODS column (Inertsil ODS-2, 5µm, 250mm x 4.6 mm ID, GL Science Inc., Tokyo, Japan) in acolumn oven at 40°C fitted with a guard column (10 mm× 4.0mm ID) using acetonitrile/ethyl acetate/0.1%phosphoric acid (85/20/895, v/v/v) as a mobile phase,flow rate at 1 ml/min, and analyzed at 280 nm with aspectrophotometric detector SPD-6A UV (Shimadzu Co.,Kyoto, Japan). Standard solutions for all catechins wereprepared in methanol. Catechin concentrations in theseaweeds were calculated using standard curve at therange of 0-50 µg/ml. A peak of each catechin wasidentified by retention time. Extraction, Hydrolysis and Analysis of FlavonoidsTotal flavonoids were extracted according to themethod of Hertog et al.19) as follows. Each minced freshseaweed sample, 5g, was homogenized with 40 ml of 75%methanol with 2g/l TBHQ (t-butylhydroquinone) using amixer (Ultra-Turrax T-25 Janke & Kunkel, GmbH Co.Staufen, Germany) at 5,000-10,000 rpm for 60 s. Tenmililiters of 6M hydrochloric acid was added andcarefully mixed. The homogenate was refluxed at 90°Cfor 2 hours, and final concentration of hydrochrolic acidwas approximately 1.2 N. After cooling, the supernatantwas filtered through an Advantec filter paper No. 101(Toyo Roshi Kaisha, Ltd., Tokyo, Japan) and transferredto a volumetlic flask with methanol. After replacing airwith nitrogen gas to inhibit decomposition of flavonoids,the extracts were kept at -80°C until analysis. The effectof hydrolysis on seaweed samples by methanol-HClmentioned above was compared with that of methanolextract. All samples were extracted and analyzed intriplicate. Flavonoids and related phenolic compounds weredetermined by high-performance liquid chromatographmodified from the methods of Hertog et al.19) and Vinsonet al.20) Flavonoids and related compounds were separatedby a C18 column (Nova-Pak C18 , 4µm, 150 mm × 3.9 mmID, Waters Co., Milford, MA, USA) fitted with a guardcolumn (20 mm × 3.9 mm ID), using 25% acetonitrile in0.025M KH2PO4 at pH 2.4 as mobile phase, flow rate at0.9 ml/min and analyzed by a diode array detector SPD-M10Avp (Shimadzu Co., Kyoto, Japan). Authenticreagents were dissolved in methanol. Flavonoidconcentration in the seaweed was calculated using acalibration curve within concentration 0-200µg/ml.Samples were separated and identified by retention timeand spectra of each peak.Results and DiscussionDistribution of the polyphenolic and relatedcompounds in two Halimeda are shown in Table 1.Contents of the polyphenolic compounds are expressed asµg of compounds per g of dry weight of seaweeds. Catechin, epicatechin, catehin gallate, epicatechingallate, epigallocatechin gallate, rutin, quercitrin, luteolin,quercetin, apigenin, kaempferol, baicalein were notdetected from both Halimeda. Quercitrin was detected inH. opuntia, but it was extremely small amount.H. macroloba and H. opuntia belong to the samespecies, and live in the same area. Their difference of theshape is quite small, however, composition ofpolyphenolics was different. H. macroloba had 28,000µgof epigallocatechin and 1,880µg of catechol, in contrast,H. opuntia had 12,700µg and 384µg of each compound.H. macroloba had caffeic acid and hesperidin, but H.opuntia did not contain caffeic acid and it had hesperidinin trace amount. In this research, while samples werecollected in the same seashore and in the same season, theresult showed the difference of polyphenolic composition.Polyphenolic contents of seaweed may have seasonal andlocal variations. Therefore, further research is required forthis field of study.Table 1. Distributions of polyphenolic compounds in Halimeda(mean ± SD  µg/g dry matter)Compounds Hiroha-sabotengusa SabotengusaHalimeda macroloba Halimeda opuntia Catechin - -Epicatechin - -Epigallocatechin 28,000 ± 6,200 12,700 ± 930Catechin gallate - -Epicatechin gallate - -Epigallocatechin gallate - -Rutin - -Caffeic acid 84.9± 21 -Catechol 1,880± 650 384± 130Quercitrin - +Hesperidin 144± 100 +Myricetin 414± 80 147± 1.2Morin 429± 4.3 234± 7.7Luteolin - -Quercetin - -Apigenin - -Kaempferol - -Baicalein - -Polyphenolic contents in Halimeda 23Terada et al.5) reported that dried tea leaves contained20,000µg of epigallocatechin, 25,000µg ofepigallocatechin gallate, 18,000µg of epicatechin gallate,and 8,000µg of epicatechin. Tea leaves from ShizuokaPrefecture, Japan, were analyzed by Suematsu et al.6), andthey reported 670µg of epigallocatechin, 220µg ofepicatechin, 740µg of epigallocatechin gallate, 140µg ofepicatechin gallate, and 35µg of catechin. Concentrationsof catechins in tea leaves varied widely according to thereports. H. macroloba was found to have a larger amountof epigallocatechin (28,000µg) than tea leaves, but exceptthat, both Halimeda had smaller amount of catechins thantea leaves. Epigallocatechin gallate, epigallocatechin,epicatechin gallate, and epicatechin were found to be themost important components in terms of antioxidant abilityamong all the catechins.19-21) All of the active catechinsmentioned above were not found in both Halimeda.Polyphenolic and related compounds occupied more than3% of dry weight H. macroloba, and such a largepercentage of polyphenolic content was also reported incase of Scutellaria baicalensis (one of the herb).21) S.baicalensis contained mainly baicalein, however,H.macroloba contained epigallocatechin instead ofbaicalein. Quercetin, luteolin and apigenin were reportedto be "anticarcinogenic" flavonoids,22) but Halimeda didnot contain them. There are free polyphenolic compoundsand conjugated one. It was reported that some vegetableshad more than 50% of conjugated form.20) From thisreport, we found the difference of polyphenoliccomposition of Halimeda, but we should find out thedifference of the structure of polyphenolic compounds,and make clear the difference of their activity on ourhealth. As Cao et al.23) mentioned the structure-activityrelationships of flavonoids, we are continuing the researchon the activities as an antioxidant or a prooxidant ofpolyphenolic compounds from seaweeds with severaloxidation models.AcknowledgementsThe authors gratefully acknowledge the efforts of Dr.Tsuyoshi Hayashibara and Mr. Kazumasa Hashimoto inIshigaki Tropical Station, Seikai National FisheriesResearch Institute, Fisheries Agency for their help incollecting samples at Ishigaki island. We also thank Dr.Jiro Tanaka and Dr. Teru Ioriya, for their help with speciesidentification. This research was partly supported by aGrant-Aid for Encouragement of Young Scientists (No.11760146) from the Ministry of Education, Science,Sports and Culture of Japan.References1) J.Y. Qian, D. Mayer, and M. Kuhn: Flavonoids infine buckwheat (Fagopyrum esculentum Monch)flour and their free radical scavenging activities.Dtsch. Lebensm.-Rundschau., 95, 343-349 (1999).2) X.J. Yan, Y. Chuda, M. Suzuki, and T. Nagata:Fucoxanthin as the major antioxidant in Hizikiafusiformis, a common edible seaweed. Biosci.Biotech. Biochem., 63, 605-607 (1999).3) H. Ohshima, Y. Yoshie, S. Auriol, and I. Gilibert:Antioxidant and pro-oxidant actions of flavonoids:Effects on DNA damage induced by nitric oxide,peroxynitrite and nitroxyl anion. Free Radic. Biol.Med., 25,1057-1065 (1998).4) T. Chan, G. Galati, and P.J. O'Brien: Oxygenactivation during peroxidase catalysed metabolismof flavonones or flavanones. Chemico-BiologicalInteractions, 122, 15-25 (1999).5) S. Terada, Y. Maeda, T. Masui, Y. Suzuki, and K.Ina: Comparison of caffeine and catechincomponents in infusion of various tea (green, oolongand black tea) and tea drinks. Nippon ShokuhinKogyo Gakkaishi, 34, 20-27 (1987). (in Japanesewith English abstract)6) S. Suematsu, Y. Hisanobu, H. Saigo, R. Matsuda,and Y. Komatsu: A new extraction procedure ofcaffeine and catechins in green tea (Studies onpreservation of constituents in canned drinks partV). Nippon Shokuhin Kagaku Kogaku Kaishi , 42,419-424 (1995). (in Japanese with English abstract)7) K. Ikegaya, H. Takayanagi, and T. Anan:Quantitative analysis of tea constituents. ChagyoKenkyu Hokoku, 71, 43-74 (1990). (in Japanese)8) M.G.L. Hertog, P.C.H. Hollman, and B. van dePutte: Content of potentially anticarcinogenicflavonoids of tea infusions, wines, and fruit juices. J.Agric. Food Chem. , 41, 1242-1246 (1993).9) G.J. Soleas, J. Dam, M. Carey, and D.M. Goldberg:Toward fingerprinting of wines: Cultivar-relatedpatterns of polyphenolic constituents in Ontariowines. J. Agric. Food Chem., 45, 3871-3880 (1997)10) A. Ghiselli, M. Nardini, A. Baldi, and C. Scaccini:Antioxidant activity of different phenolic fractionsseparated from an Italian red wine. J. Agric. FoodChem., 46, 361-367 (1998).11) C. Sanbongi, N. Osakabe, M. Natsume, T. Takizawa,S. Gomi, and T. Osawa: Antioxidative polyphenolsisolated from Theobroma cacao. J. Agric. FoodChem., 46, 454-457 (1998). Y. Yoshie, W. Wang, Y. Hsieh and T. Suzuki2412) N. Osakabe, M. Yamagishi, C. Sanbongi, M.Natsume, T. Takizawa, and T. Osawa: Theantioxidative substances in cacao liquor. J. Nutr. Sci.& Vitaminol., 44, 313-321 (1998).13) I.C.W. Arts and P.C.H. Hollman: Optimization of aquantitative method for the determination ofcatechins in fruits and legumes, J. Agric. FoodChem., 46, 5156-5162 (1998).14) B. Risch and K. Herrman: Die Gehalte anHydroxyzimtsaeure- Verbindungen und Catechinenin Kern- and Steinobst (Contents ofhydroxycinnamic acid derivatives and catechins inpome and stone fruit). Z. Lebensm. Unters. Forsch.,186, 225-230 (1988).15) C. Quesada, B. Bartolome, O. Nieto, C. Gomez-Cordoves, T. Hernandez, and I. Estrella: Phenolicinhibitors of (-amylase and trypsin enzymes byextracts from pears, lentils, and cocoa. J. FoodProtection, 59, 185-192 (1996) 16) S.H. Hakkinen, S.O. Karenlampi, I.M. Heinonen,H.M. Mykkanen, and A.R. Torronen: Content offlavonols quercetin, myricetin, and kaempferol in 25edible berries. J. Agric. Food Chem., 47, 2274-2279(1999).17) M.G.L. Hertog, P.C.H. Hollman, and M.B. Katan:Content of potentially anticarcinogenic flavonoidsof 28 vegetables and 9 fruits commonly consumedin the Neterlands. J. Agric. Food Chem., 40, 2379-2383 (1992).18) Y. Yoshie, W. Wang, D. Petillo, and T. Suzuki:Distribution of catechins in Japanese seaweeds.Fisheries Sci., 66, 998-1000 (2000).19) M.G.L. Hertog, P.C.H. Hollman, and D.P. Venema:Optimization of a quantitative HPLC determinationof potentially anticarcinogenic flavonoids invegetable and fruits. J. Agric. Food Chem., 40, 1591-1598 (1992).20) J.A. Vinson, Y. Hao, X. Su, and L. Zubik: Phenolantioxidant quantity and quality in foods:Vegetables. J. Agric. Food Chem., , 46, 3630-3634(1998).21) M.C. Lin, M.J. Tsai, and K.C. Wen: Supercriticalfluid extraction of flavonoids from ScutellariaeRadix. J. Chromatogr., , 830, 387-395 (1999).22) P.C.H. Hollman, and M.B. Katan: Dietaryflavonoids: Intake, health effects and bioavailability.Food & Chemical Toxicology, 37, 937-942 (1999).23) G. Cao, E. Sofic, and R.L. Prior: Antioxidant andprooxidant behavior of flavonoids: structure-activityrelationships. Free Radic. Biol. Med., 22, 749-760(1997).サボテングサ 2種のポリフェノール類の含量吉江由美子・王　 ・謝　雅珮・鈴木　健（東京水産大学食品生産学科）ポリフェノール化合物は、脂質やDNAの酸化に対して影響力を持つとされている。本研究では沖縄で同時期に採取されたヒロハサボテングサならびにサボテングサを試料とし、ポリフェノール化合物16種および関連化合物としてカテコール、カフェイン酸を、ODSカラム、C18カラムとダイオードアレイ検出器を用いたHPLCによって分析した。ヒロハサボテングサにのみカフェイン酸が検出された。ヒロハサボテングサからはサボテングサでごく微量であったヘスペリジンが検出された。ヒロハサボテングサのカテコール含量はサボテングサの5倍ほどであった。エピガロカテキン、ミリセチンとモリンはヒロハサボテングサにおいてサボテングサの約2倍の含量が認められた。このように、類似した海藻2試料から異なる含量のポリフェノールならびに関連化合物が検出された。キーワード：ポリフェノール化合物、サボテングサ、HPLC-ダイオードアレイ分析烟�韋

サボテングサ2種のポリフェノール類の含量

https://oacis.repo.nii.ac.jp/?action=repository_action_common_download&item_id=106&item_no=1&attribute_id=22&file_no=1

サボテングサ2種のポリフェノール類の含量

Abstract

Similar works

Full text

Available Versions

Open Access Collection of International and Scholarly Papers