한수지 49(2), , 2016 Original Article Korean J Fish Aquat Sci 49(2), ,2016 전기투석시스템을활용한염생식물칠면초 (Suaeda japonica) 추출물의염분표준화에따른주요성분및생리활성비교연구 박현

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1 한수지 49(2), 243, 6 Original Article Korean J Fish Aquat Sci 49(2),243,6 전기투석시스템을활용한염생식물칠면초 (Suaeda japonica) 추출물의염분표준화에따른주요성분및생리활성비교연구 박현호,2,3 고석천 3 정원교,2,3 부경대학교의공학과, 2 부경대학교 BK2 Plus 해양융복합생명의학기술센터, 3 부경대학교해양융복합바이오닉스연구단 Comparison of the Biological Activities of Electrodialysisdesalted Bioactive Compounds from the Halophyte Suaeda japonica HyeonHo Park,2,3 SeokChun Ko 3 and WonKyo Jung,2,3 Department of Biomedical Engineering, Pukyong National University, Busan 4853, Korea 2 Centre for MarineIntegrated Biomedical Technology (BK2 Plus), Pukyong National University, Busan 4853, Korea 3 MarineIntegrated Bionics Research Center, Pukyong National University, Busan 4853, Korea To effectively remove salts from Suaeda japonica, extracts, an electrodialysis system was developed. The biological activities of nondesalted () and desalted S. japonica () extracts were compared. The extract exhibited superior polyphenolic (6.26%) and carbohydrate (28.56%) contents. The IC 5 values of the extract against DPPH radicals and hydrogen peroxide were.22 and.68 mg/ml, respectively, which was higher than that of the extract. Neither the nor the extract was cytotoxic in RAW macrophages. Additionally, the extract had a higher NO inhibitory effect compared to the extract in lipopolysaccharide (LPS)activated RAW macrophages. These data indicate that extracts have greater biological activity than do ND extracts, and application of the electrodialysis process may be useful in marine bioresource applications. Key words: Suaeda japonica, Desalting, Electrodialysis system, Biological activity 서론,, (Lee et al., 2). (Kong et al., 8). 2,,. (Suaeda japonica) (Chenopodiaceae) SinoJapanese group, (Cho et al., 4). (Lee et al., 2; Lee et al., ), (Kang et al., 3) (Kim et al., ) (Choi et al., ; Choi et al., 9).,., (osmolarity),, ph., (hypertension) Korean J Fish Aquat Sci 49(2) 243, April 6 This is an Open Access article distributed under the terms of the Creative Commons Attribution NonCommercial Licens ( which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Received 2 January 6; Revised 3 March 6; Accepted 4 March 6 Corresponding author: Tel: Fax: address: wkjung@pknu.ac.kr Copyright 6 The Korean Society of Fisheries and Aquatic Science 24 pissn:3748, eissn:

2 염생식물칠면초의염분표준화에관한연구 25 (hypernatremia) (Jung and Shim, 8; Kim, 6). (cell death) (cell apoptosis), (Michea et al., ; Stela, 994). (distillation), (reverse osmosis), (vapor compression) (electrodialysis) (Cho and Han, ). (ED). Maigrot and Savates Meyer and Strauss, Juda and McRae,,, (Park et al., )..,. 시약 재료및방법,diphenyl2pricylhydrazyl (DPPH), 2deoxyribose, peroxidase, 2,2 azinobis(3ethylbenzthiazoline)6sulfonic acid (ABTS), FolinCiocalteu reagent, Dimethyl sulfoxide (DMSO), Griess reagent (% sulfanilamide and.% naphthylethylenediamine dihydrochloride in 2.5% phosphoric acid), 3[4,5dimethylthiazol2yl]2,5diphenyltetrazoliumbromide (MTT) Sigma Co. (St. Louis, MO, USA). Dublecco s modified eagle s medium (DMEM), fetal bovine serum (FBS) phosphate buffer saline (PBS) Gibco BRL Co. (Gaithersburg, MD, USA).. 칠면초 4. ( 칠면초추출물조제 7% : (w/w) , rpm.,.. 전기투석장치 (CJS3, ),, (B ) (A ). B A 2., (Fig. ). (C ) 5% (w/v) Na 2 SO 4 5 ml,, ml 3 g 5 ml (AC, CHANG JO, SEOUL, KOREA) 5, 5 V. Da,.,,. A B C Fig.. Conceptual flow diagram of electro dialysis system. A, desalination tank; B, Concentrate tank; C, electrode tank.

3 26 박현호ㆍ고석천ㆍ정원교 총 Polyphenol 함량측정 ( ) polyphenol Singleton et al. (999). L 7.5% sodium carbonate (Na 2 CO 3 ) 25 L 5. N FolinCiocalteu reagent 3 L 3 75 nm. gallic acid polyphenol. 총당함량측정 Dubois et al. (956). L 5% phenol L H 2 SO 4 5 L 49 nm. glucose. DPPH radical 소거활성측정 DPPH radical Nanjo et al. (996). 96well plate L.5 4 M DPPH L 3 57 nm. DPPH radical. DPPH radical (%)=[( )/ ] Hydrogen peroxide 소거활성측정 hydrogen peroxide Müller (985).. M phosphate buffer (ph 5.) L 96well plate L hydrogen peroxide mm ABTS peroxidase ( unit/ml) 3 L 37 5 nm. Hydrogen peroxide. Hydrogen peroxide (%)=[( )/ ] 세포배양 RAW American Type of Culture Collection (ATCC, Rockville, VA, USA). % FBS % streptomycin/penicillin (Gibco/BRL, Gaithersburg, MD, USA) DMEM 5% CO 세포독성평가 MTT assay. 5 cells/ml 96well plate 5% CO ,,, g/ml. lipopolysaccharide (LPS, g/ml) L MTT well formazan 4 MTT L DMSO 5 nm. Nitric oxide (NO) 생성억제활성측정 5 cells/ml 24well plate 5% CO ,,, g/ml. lipopolysaccharide (LPS, g/ml) 24. L Griess reagent L, 5 nm. 통계분석 (Mean) (standard deviation). SPSS (SPSS Inc. Ver.2.) Oneway ANOVAtest Duncan s multiple range test (P<.5). 결과및고찰 탈염후전기전도도, 염함량및수율,, (Fig. 2). (conductivity). (Table ). 8.9 ms/cm.8 ms/cm, 9 mm 3 mm., 53.33%.

4 염생식물칠면초의염분표준화에관한연구 27 Diluate Diluate Anode Cathode Fig. 2. Principle of ionexchange membrane of electro dialysis system.,., (Lee et al., 2).. 탈염전후의총폴리페놀및총당함량변화 Concentrate Concentrate Table.Change of condition and yield after desalting by electro dialysis system Conditions Before Desalting After Conductivity (ms/cm) 8.9±..±. NaCl (M).9±..2±. Yield (%) 53.33±.53 P <.5 indicates significantly different from nondesalting. 2,, (Kang et al., 4; Kim et al., ; Kim et al., 3). Table 2.Change of polyphenolic and carbohydrate contents after desalting by electro dialysis system Components Before Desalting After Total polyphenol (%) 4.48± ±. Total carbohydrate (%) 5.56± ±.7 P <.5 indicates significantly different from nondesalting.. Table % 6.26%.8%, 5.56% 28.56% 3%.. DPPH radical 소거활성 (Park et al., 2).. Free radical, DPPH radical free radical,. radical (Ko et al., a; Yokozawa et al., 988). DPPH radical Fig. 3..,.2.4 mg/ml, DPPH radical. DPPH radical.2.4 mg/ml DPPH radical scavenging activity (%)..2.4 IC 5 value (mg/ml) Concentration (mg/ml) Fig. 3. DPPH radical scavenging activities of and from Suaeda japonica. Values are expressed as means ± standard deviation (SD) in triplicate experiments. Statistical evaluation was performed to compare the and. P< ± ±.3

5 28 박현호ㆍ고석천ㆍ정원교, 5% (IC 5 value).35 mg/ml.22 mg/ml. DPPH radical (Kim et al., ; Oki et al., 2),. Hydrogen peroxide 소거활성 Hydrogen peroxide non free radical (Reactive oxygen species) DNA,, (Ko et al., b; Park and Kang, 2). hydrogen peroxide Fig ,.5 mg/ml, hydrogen peroxide. hydrogen peroxide.5 mg/ml, 5% (IC 5 value) mg/ml,.68 mg/ml. DPPH radical., hydrogen peroxide Lee et al. (2) hydrogen peroxide. Kim Hydrogen peroxide scavenging activity (%).25.5 IC 5 value (mg/ml) Concentration (mg/ml) Fig. 4. Hydrogen peroxide scavenging activities of and from Suaeda japonica. Values are expressed as means ± standard deviation (SD) in triplicate experiments. Statistical evaluation was performed to compare the and. P<.5. >.6 ±.3 et al. (3).. 세포독성확인 LPS MTT., LPS LPS g/ml (Fig. 5). NO g/ml. NO 생성억제능확인,,, (Heo et al., ). (cytokine) (Ko and Jeon, 5). NO NO (NOS) (arginine), (Kim et al., 999)., NO, NO (Ko and Jeon, 5). NO LPS., LPS NO %, Cell viability (%) Extract (μg/ml ) LPS ( μg/ml ) Fig. 5. Cytotoxic effects of and from S. japonica on viability in RAW macrophages. Cells were treated with and from Suaeda japonica at the indicated concentrations (,, and μg/ml). Values are expressed as means ± standard deviation (SD) in triplicate experiments.

6 염생식물칠면초의염분표준화에관한연구 29 NO production (%) Extract (μg/ml ) LPS ( μg/ml ) Fig. 6. Inhibitory effects of and from Suaeda japonica on LPSstimulated NO production in RAW macrophages. Production of NO was assayed in the culture medium of macrophages stimulated with LPS ( μg/ml) for 24 h in the presence of and from S. japonica (,, and μg/ml). Values are expressed as means±standard deviation (SD) in triplicate experiments. Statistical evaluation was performed to compare the and. P<.5 vs LPStreated group and sampletreated group, as indicated. P<.5 vs and, as indicated. g/ml,, g/ml., g/ml.,. 2.,., (, ) (, ).,.,,,... 사사 3 (52). References Cho BY and Han YW.. A study on Seawater desalination using EDR method. Korean J Soc Water Wastewater 5, Cho JY, Huang Z, Park SY, Park KH, Pai TK, Kim SY, Kim HR and Ham KS. 4. The effects of several halophytes on insulin resistance in otsuka longevans tokushima fatty rats. Korean J Food Sc Technol 46, 7. org/.972/kjfst Choi JI, Kim YY, Kim JH, Kwon JH, Ahn DH, Chun BS and Lee JW.. Physiological activities of Suaeda japonica extracts on harvest season. Korean J Soc Food Sci Nutr 39, Choi JI, Kim YJ, Kim JH, Song BS, Yoon Y, Byun MW, Kwon JH, Chun SS and Lee JW. 9. Antioxidant activities of the extract fractions from Suaeda japonica. Korean J Soc Food Sci Nutr 38, jkfn Dubois M, Gilles KA, Hamilton JK, Rebers PA and Smith F Colorimetric method for determination of sugar and related substances. Anal Chem 28, org/.2/aca7 Heo SJ, Yoon WJ, Kim KN, Ahn GN, Kang SM, Kang DH, Affan A, Oh C, Jung WK and Jeon YJ.. Evaluation of antiinflammatory effect of fucoxanthin isolated from brown algae in lipolysaccharidestimulated RAW macrophages. Food Chem Toxicol 48, org/.6/j.fct..5.3 Jung CE and Shim EG. 8. Saltrelated dietary behaviors and sodium intakes of university students in Gyeonggido. Korean J Soc Food Sci Nutr 37, org/.3746/jkfn Kang H, Koppula S, Kim HK and Park TK. 3. Suaeda japonica Makino attenuates lipopolysaccharideinduced neuroinflammatory responses in BV2 microglia via NFkappa B signaling. Trop J Pharm Res 2, org/.434/tjpr.v2i3.2 Kang MC, Kim SY, Kim YT, Kim EA, Lee SH, Ko SC, Wijesinghe WA, Samarakoon K W, Kim YS, Cho JH, Jang HS and Jeon YJ. 4. In vitro and in vivo antioxidant activities of polysaccharide purified from aloe vera (Aloe bardadensis) gel. Carbohydrate Polymers 99, dx.doi.org/.6/j.carbpol Kim DU. 6. Hypernatremia and hyponatremia. Korean J Pediatrics 49, Kim HK, Cheon BS, Kim YH, Kim SY and Kim HP Ef

7 3 박현호ㆍ고석천ㆍ정원교 fects of naturally occurring flavonoids on nitric oxide production in the macrophage cell line RAW and their structureactivity relationships. Biochem Pharmacol 58, Kim HH, Jun BS, Kim SK, Cha JY and Cho YS.. Polyphenolic compound content and antioxidative activities by extracts from seed, sprout and flower of safflower (Carthamus tinctiorius L.). Korean J Soc Food Sci Nutr 29, Kim JG, Kang YM, Eum GS, Ko YM and Kim TY. 3. Antioxidative activity and antimicrobacteral activity of extracts from medicinal plants (Akebia quinate Decaisn, Scirusfluviatilis A. Gray, Gardenia jasminoides for. Grandiflora Makino). J Agric Life Sci 37, Kim J, Song JY, Lee JM, Oh SH, Lee HJ, Choi HJ, Go JH and Kim YH.. A study on physicochemical property of Salicornia herbaciea and Suaeda japonica. J Fd Hyg Safety 25, 779. Ko SC and Jeon YJ. 5. Antiinflammatory effect of enzymatic hydrolysates from Styela clava flesh tissue in lipopolysaccharidestimulated RAW macrophages and in vivo zebrafish model. Nutr Res Prac 9, dx.doi.org/.462/nrp Ko SC, Kang SM, Ahn G, Yang HP, Kim KN and Jeon YJ. a. Antioxidant activity of enzymactic extracts from Sargassum coreanum. Korean J Soc Food Sci Nutr 39, Ko SC, Kang SM., Lee SH, Ahn G, Kim KN, Kim YT, Kim JS, Heu MS and Jeon YJ. b. Protective effect of enzymatic extracts from Sargassum coreanum on H 2 O 2 induced cell damage. Fish Aquat Sci 3, org/.5657/fas Kong CS, Um YR, Lee JI, Kim YA, Lee JS and Seo YW. 8. Inhibition effects of extracts and its solvent fractions isolated from Limonium tetragonum on growth of human cancer cells. Korean J Biotechnol Bioeng 23, Lee KS, Kim AJ and Lee KY. 2. Analysis of Chemical Composition and Antioxidant Activity of Suaeda japonica. J East Asian Soc Dietary Life 22, org/.3746/jkfn Lee KS, Gim JC, Son SM and Lee KY.. Antioxidative effect of Suaeda japonica ethanol extract and solvent partitioned fractions. Korean J Soc Food Sci Nutr, Michea L, Ferguson D R, Peters E M, Andrews P M, Kirby M R and Burg M B.. Cell cycle delay and apoptosis are induced by high salt and urea in renal medullary cells. Am J Physiol Renal Physiol 278, 928. Müller HE Detection of hydrogen peroxide produced by microorganism on ABTSperoxidase medium. Zentrabl Bakteriol Mikrobiol Hyg 259, org/.6/s766724(85)456 Nanjo F, Goto K, Seto R, Suzuki M, Sakai M and Hara Y Scavenging effects of tea catechins and their derivatives on,diphenyl2picrylhydrazyl radical. Free Radic Biol Med 2, (96)2377 Oki T, Masuda M, Furuta S, Nishibia Y, Terahara N and Suda I. 2. Involvement of anthocyanins and other phenolic compounds in radicalscavenging activity of purplefreshed sweet potato cultivars. Food Chem Toxicol 67, Park EJ and Kang MH. 2. Application of the alkaline comet assay for detecting oxidative DNA damage in human biomoditoring. Korean J Nutr 35, Park HJ, Kim MM and Oh YH. 2. Effect of fruit extract of Prunus mume on the scavenging activity of reactive oxygen species and melannin production in B6F cells. J Life Science 22, JLS Park PJ, Lee SH and Kim SK.. Desalination of boiled oyster extract by electrodialysis. Korean J Biotechnol Bioeng 5, Singleton VL, Orthofer R and LamuelaRaventos RM Analysis of total phenols and other oxidation substrates and antioxidants by means of folinciocalteu reagent. Methods Enzymol 299, Stela H Oxidation of ascorbic acid and metal ions as affected by NaCl. J Agric Food Chem 42, dx.doi.org/.2/jf47a8 Yokozawa T, Chen CP, Dong E, Tanaka T, Nonaka GI and Nishioka I Study on the inhibitory effect of tannins and flavonoids against the, diphenyl2picrylhydrazyl radical. Biochem Pharm 56, S62952(98)282

제 출 문 경상북도 경산시 농업기술센터 귀하 본 보고서를 6차산업수익모델시범사업 농산물가공품개발 연구용역 과제의 최종보고서로 제출합니다. 2014 년 11 월 19 일 주관연구기관명 : 영남대학교 총괄연구책임자 : 한 기 동 연 구 원 : 김 상 욱 이 수 형 이 상

6차 산업 수익모델 시범사업 농산물가공품개발 연구용역 최 종 보 고 서 영남대학교 식품공학과 연구책임자 : 한 기 동 제 출 문 경상북도 경산시 농업기술센터 귀하 본 보고서를 6차산업수익모델시범사업 농산물가공품개발 연구용역 과제의 최종보고서로 제출합니다. 2014 년 11 월 19 일 주관연구기관명 : 영남대학교 총괄연구책임자 : 한 기 동 연 구 원 : 김