J Korean Soc Food Sci Nutr 한국식품영양과학회지 42(12), 1893~1898(2013) http://dx.doi.org/10.3746/jkfn.2013.42.12.1893 해조류에탄올추출물의항산화및 Tyrosinase 억제활성 이나영 군산대학교식품생명공학과 Antioxidant Effect and Tyrosinase Inhibition Activity of Seaweeds Ethanol Extracts Na Young Lee Dept. of Food Science and Biotechnology, Kunsan National University, Jeonbuk 573-701, Korea ABSTRACT Seaweeds, laver, sea mustard, kelp, and fusiformis, were prepared and investigated for its antioxidant and tyrosinase inhibition activities. The extracts yield, color, total phenolic contents, antioxidative activity, and tyrosinase inhibition activity of the extract samples were measured. Hunter Lightness values of laver, sea mustard, kelp, and fusiformis extracts were 82.88, 78.53, 83.04, and 78.11, respectively. The contents of total phenolic compounds of the seaweed extracts powder, laver, sea mustard, kelp, and fusiformis were 43.23, 11.59, 10.09, and 46.59 mg/g of sample, respectively. 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of the fusiformis extract was shown to be the highest value compared with other seaweed extracts. 2,2'-azino-bis-3-ethylbenzo-thiazoline-6-sulfonic acid (ABTS) radical scavenging activities of laver, sea mustard, kelp, and fusiformis extracts were 258.00, 219.26, 95.77, and 1186.62 µmol trolox equivalence per gram, respectively, at the 1,000 ppm level. TBARS value of oil emulsion, samples without extracts was higher than those of the samples prepared with laver and sea mustard extracts. The inhibition rates (%) of the mushroom tyrosinase of laver, sea mustard, kelp, and fusiformis extracts powder were 25.93, 26.32, 24.76 and 20.24% at 1,000 ppm, respectively. The results indicated that laver, sea mustard, kelp and fusiformis extracts possess biological activities such as antioxidant activity and tyrosinase inhibition effect. Key words: seaweed extracts, antioxidant activity, tyrosinase inhibition activity 서 최근소비자들이건강에대한인식이증가하면서건강기능식품에대한관심이높아지고있다. 이에따라건강기능식품개발을위해건강관련기능성물질탐구, 기능성원료및식품개발에대한연구가꾸준히진행되고있고, 기능성소재개발을위해천연물, 농수산물및약용식물로부터생리활성평가및물질분리에대한연구가활발히진행중에있다. 해조류 (seaweed) 는해양에서서식하는거대조류 (macroalgae) 로서바다에생육하는다세포원생생물을지칭하며육안으로관찰할수있는크기를가진점에서단세포의미세조류 (microalgae) 와구별된다. 해조류내의구분은광합성색소에따라녹조류 (green seaweed), 갈조류 (brown seaweed), 홍조류 (red seaweed) 로나눠지며각각이지니는서식환경및구성성분에서도차이를보인다. 또한세계적해조류생산은주로양식에의해이루어지고있는데대부분의수요는아시아지역을중심으로이루어져전체소비량 론 Received 22 August 2013; Accepted 8 November 2013 E-mail: nylee@kunsan.ac.kr, Phone: 82-63-469-1826 의 83~90% 를차지하고있다. 우리나라는중국, 필리핀, 일본에이어세계 4위의해조류생산국으로서생산량은 2008 년기준 93만톤이며, 이는전체어업생산량의 27.8% 및양식생산량의 67.6% 를차지하고있다 (1). 우리나라의해조류는주로김, 미역, 다시마, 톳및우무등이식용으로이용되고있으며, 가공형태로는건제품, 염장품및조미품등으로제조되거나식품의부재료로이용되기도한다. 해조류의영양성분은열량이낮고식이섬유, 비타민, 무기질, 미네랄등이풍부하여유용식용자원중의하나이다 (2,3). 세계건강기능식품시장규모는점차증가하는추세이며건강기능식품을포함한소위국제영양산업 (nutrition industry) 시장은규모가증가하고있다. 또한국내의경우도건강기능식품시장은다양한융 복합기술의발달에따라새롭고다양한건강기능식품발굴이늘어날것으로전망하고있다 (4). 최근해조류에함유된탄수화물이장관운동을원활히하고중금속배출을촉진시키며고지혈증의개선에유효하다는결과가발표되면서해조류를이용한건강기능식품개발에대한관심이높아지고있다 (5-7). 이와더불어해조류수용성식이섬유의혈압강하, 콜레스테롤저하등의효과가
1894 이나영 보고되고있으며 (8,9), 해조류의다당류인퓨코이단의혈중지질억제, 종양세포의성장저해활성이보고되면서기능성을갖는해조다당류의추출, 분리에대한연구가꾸준히진행되고있다 (10-12). 또한해조류추출물이 acetylcholinesterase 저해활성에효과를보여치매치료제로서의가능성이보고되었다 (3). 따라서본연구에서는기능성소재개발을위해국내에서주로생산및소비되는주요해조류인김, 미역, 다시마및톳을이용하여제조한추출파우더의항산화활성, 지질산패억제및 tyrosinase 저해활성을확인하여건강기능식품개발을위한기초자료로활용하고자하였다. 재료및방법시료제조실험에사용된김 (Porphyra tenera), 미역 (Undaria pinnatifida), 다시마 (Laminaria japonica Areschoug) 및톳 (Hizikia fusiformis) 은시중에서구입하여실험에사용하였다. 구입한시료는추출을위해 2 2 cm로자른후 70% 에탄올을이용하여실온에서 24시간동안추출을진행하였으며, 추출시사용된시료와용매는 1:20(w/v) 비율로추출하였다. 추출종료후 110 nm 여과지 (No. 2, Advantec, Toyo Co., Tokyo, Japan) 를이용하여여과한후여액은진공농축기 (Eyela New Rotary Vacuum Evaporator, Rikakikai Co., Tokyo, Japan) 를이용하여농축하였다. 농축된시료는진공동결건조기 (Eyela FD1, Rikakikai Co.) 를이용하여건조하였으며건조된시료의수율을측정하였다. 건조된시료는 -20 C 냉동고에서보관하면서실험에사용하였다. 시약분석에사용되어진시약인 1,1-diphenyl-2-picrylhydrazyl radical(dpph), Folin Ciocalteu's phenol reagent, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid(trolox), 2,2'-azino-bis-3-ethylbenzothiazoline- 6-sulfonic acid(abts), L-3,4-dihydroxyphenylalanine, mushroom tyrosinase, gallic acid 등은 Sigma-Aldrich Co.(St. Louis, MO, USA) 로부터구입하여실험에사용하였다. Folin-Ciocalteau 시약 500 μl 및 20% Na 2CO 3 용액 1.5 ml를가한후 23 C에서 2시간정치한후분광광도계 (UV- 1800, Shimadzu Co., Kyoto, Japan) 를이용하여 765 nm 에서흡광도를측정하였다. 총페놀함량은 gallic acid를이용하여작성한표준검량곡선으로환산하여산출하였다. 라디칼소거능라디칼소거능은 DPPH 및 ABTS 라디칼소거능에대해측정하였다. DPPH 라디칼소거능은 Blois 방법 (14) 에준하여측정하였다. 시료추출물 (1,000 ppm, 1 ml) 에 0.2 mm DPPH 라디칼용액을첨가한후혼합하여실온에서 30분동안반응시켰으며, 반응액은 517 nm에서흡광도를측정하였다. 각시료의라디칼소거능은다음식에의해계산하여백분율로나타내었다. DPPH 라디칼소거능 (%)=[1 시료첨가구의 O.D./ 무처리구의 O.D.] 100 ABTS 라디칼소거능은 Zhao 등 (15) 의방법을이용하여측정하였다. ABTS 7 mm과 potassium persulphate 2.45 mm을하루동안암소에방치하여 ABTS + 양이온을형성시킨후 30 C에서온도평형을실시한후 734 nm에서흡광도를측정하여값이 0.70(±0.02) 이되도록에탄올로희석하였다. 희석된 ABTS + 용액 2.9 ml에추출물 (1,000 ppm, 0.1 ml) 을첨가한후혼합하여 30 C에서 20분간반응시켜 734 nm에서흡광도를측정하였다. ABTS 라디칼소거능 (%) 은검량곡선을이용하여 Trolox(TE μmol/g) 함량으로산출하였다. 지질산패도 (2-thiobarbituric acid reactive substances value, TBARS) 측정 TBARS는 Ahn 등 (16) 의방법에준하여측정하였다. 증류수에대두유 1% 및 triton X-100을첨가하여균질화과정을거쳐 oil 에멀젼을제조하였다. 에멀젼에시료를첨가한후 50 C에서 3일동안저장하면서시료의지질산패도를분석하였다. 오일에멀젼에시료를첨가한샘플을균질시키고, 균질된시료에 2-thiobarbituric acid(tba)/trichloroacetic acid(tca) 용액 (20 mm TBA in 15% TCA) 을첨가하여혼합한후 90 C 수조에서 15분간반응후얼음물에서 10분간냉각하였다. 반응용액을 2,000 g에서 15분간원심분리한후상등액은 532 nm에서흡광도를측정하였다. 색도측정실험에사용한추출용액의색도는색차계 (Color JS55, Color Technology System Co., Tokyo, Japan) 를이용하여측정하였으며각시료당 3회반복하여측정하였다. 추출물의색도는 Hunter L*, a*, b* 값으로나타냈다. 총페놀함량총페놀함량은 Yu 등 (13) 의방법을이용한 Folin-Ciocalteau 비색법으로측정하였다. 추출물 (1%, 100 μl) 에 Tyrosinase 억제활성 Tyrosinase 저해활성은 Jo 등 (17) 의방법을이용하여측정하였다. 시료 (1,000 ppm, 0.2 ml) 에 10 mm L-3,4-dihydroxyphenylalanine(L-DOPA), 50 mm potassium phosphate buffer(ph 6.5), mushroom tyrosinase(172 unit/ml, Sigma-Aldrich Co.) 를첨가혼합하여 25 C에서 15분간반응시킨후얼음물에서 15분간정치시켰다. 반응에서생성된 dopachrome은분광광도계를이용하여 475 nm에서흡광도를측정하였다. Tyrosinase 저해활성은시
해조류추출물의생리활성 1895 료용액첨가구와무첨가구의흡광도감소율을백분율로나타내었다. 통계분석통계처리는 SAS(statistical analysis system) 통계 package(version 7.0, SAS Institute, Cary, NC, USA) 를이용하여평균및표준오차를구하였으며, ANOVA 분석후다중비교법 (Duncan's multiple range test) 으로 P<0.05에서유의차검정을실시하였다. 각분석항복간의유의적상관관계를분석하기위하여총페놀함량, DPPH 및 ABTS 라디칼소거활성, tyrosinase 억제활성값의분석은 P< 0.0001 확률수준에서피어슨상관관계분석을실시하였다. Table 1. Color changes in Hunter color value of sea algae extracts Sample L * a * b * 결과및고찰 추출수율및색도분석우리나라에서주로소비되는해조류인김, 미역, 다시마및톳을이용하여추출물을제조하였고추출물의항산화활성, 지질산패억제능및 tyrosinase 억제활성을확인하였다. 70% 에탄올을이용하여추출한김, 미역, 다시마및톳추출물의수율은 3.00, 5.38, 11.73 및 0.72% 를나타내는것으로확인하였다. 식품의색은음식선택시고려되는중요한특성중의하나이며, 식품의선택기준이된다 (18). 다양한생리활성을갖는추출물을이용하여가공식품을제조할경우추출물의색도는제품의색도에영향을미칠수있다. 해조류를이용하여추출한추출물의색도를분석하였으며그결과는 Table 1과같다. 김, 미역, 다시마및톳추출물의명도는 82.88, 78.53, 83.04 및 78.11로다시마추출물의명도가가장밝은것으로나타났으나김추출물과유의적차이는나타나지않았다. Kim 등 (19) 은해조류분말제조시건조과정중의온도는해조분말의갈변현상을일으켜분말의색도에영향을미친다고보고하였다. Kim 등 (20) 은다시마추출액을이용한발효음료개발을위해 tea fungus로부터분리한 Gluconacetobacter 속을접종하여발효한다시마발효액의색도분석결과명도는 92.17~99.17로투명도가매우높았고적색도는유의적으로 green쪽에더가까웠다고보고하였다. 또한 Jeon과 Choi(21) 는해조류를첨가한돈육패티의 L* 값은해조분말첨가에따라감소하였으며톳첨가패티에서가장 82.88±1.04 a 78.53±1.77 b 83.04±2.34 a 78.11±1.31 b -3.34±0.18 a -11.95±0.06 b -3.14±0.26 a -3.31±0.20 a 11.91±0.45 c 26.77±0.83 a 7.99±0.72 d 19.80±0.60 b a-d Different letters within the same column differ significantly (P<0.05). Tabel 2. Total phenolic compounds of sea algae extracts powders Sample Total phenolic compound (mg/g) 42.23±0.98 b 11.59±0.12 c 10.09±0.06 c 46.59±1.95 a a-c Different letters within the same column differ significantly (P<0.05). 낮고파래와미역첨가패티순으로낮았다고보고하였다. 총페놀함량분석 해조류추출물의총페놀함량을분석하였으며그결과는 Table 2와같다. 김, 미역, 다시마및톳의총페놀함량은각각 42.23, 11.59, 10.09 및 46.59 mg/g으로나타났으며톳추출물이가장높은총페놀함량을나타내는것으로확인되었다. Kim 등 (19) 은다시마및미역추출물의폴리페놀을측정한결과각각 3.91 및 3.55 mg/g을나타냈다고보고하였다. 또한 Kim 등 (22) 은구멍쇠미역과괭생이모자반의총페놀함량이각각 10.84 mg/g과 7.22 mg/g으로높았으며전자공여능과 hydroxy radical 소거능도 71.40% 와 83.98% 를나타냈다고보고하였다. Ahn 등 (2) 은 35종해조류추출물의성분분석결과총 flavonoid, 총 polyphenol, 총당및환원당함량은해조류종류에따라다양하였으며갈조류는총 flavonoid, 총 polyphenol 함량이상대적으로높으며, 홍조류와녹조류는상대적으로총당함량이높은결과를나타냈다고보고하였다. 해조류추출물의항산화활성해조류추출물의라디칼소거능을분석하였으며라디칼소거능의경우 DPPH 및 ABTS 라디칼함량을분석하였다 (Table 3). DPPH 라디칼의경우톳추출물이 82.43% 로가장높은라디칼소거능을나타냈으며김추출물의경우 26.76% 의라디칼소거능을확인하였다. ABTS 라디칼소거능의경우 DPPH 라디칼소거능과유사한경향을나타냈다. 톳추출물의 DPPH 라디칼소거능은 Trolox 함량기준으로 1,186.62 μm/g의라디칼소거능이나타나는것으로확인하였다. Jeon 등 (3) 은김및톳에탄올추출물을제조한후 Table 3. DPPH and ABTS radical scavenging acitivity of sea algae extracts powders Sample DPPH radical scavenging activity (%) 26.76±1.02 b 1.84±1.31 c 4.87±0.56 c 82.43±0.45 a ABTS radical scavenging activity (Trolox μm/g) 258.00±11.10 b 219.26±13.75 b 95.77±11.68 c 1,186.62±31.03 a a-c Different letters within the same column differ significantly (P<0.05).
1896 이나영 Table 4. Thiobarbituric acid reactive substances (TBARS) value of sea algae extracts powders Sample Control TBA values 1 day 2 days 3 days 0.02±0.01 d 0.17±0.03 b 0.16±0.02 a 0.01±0.01 e 0.04±0.01 d 0.13±0.01 a 0.13±0.01 a 0.13±0.01 c 0.15±0.01 a 0.06±0.01 c 0.11±0.01 c 0.84±0.67 a 0.07±0.01 b 0.23±0.02 a 0.27±0.03 a a-e Different letters within the same column differ significantly (P<0.05). 0 이의 DPPH 라디칼소거능을확인한결과김및톳의경우각각 47.4 및 13.3% 의라디칼소거능을나타낸다고보고하였다. Kim 등 (19) 은미역과다시마의물추출파우더의 DPPH 라디칼소거능을확인한결과 10% 추출물의경우 11.20% 의라디칼소거능을확인하였으며대조구로 BHA(1 mg/ml) 는 34.32% 의라디칼소거활성을나타냈다고보고하였다. 이는본연구에사용된추출물과비교했을때대조구로사용된 BHA보다톳추출물이보다높은라디칼소거능을나타내는것으로확인하였다. 해조류추출물의지질산화억제능분석해조류추출물을첨가하여제조된오일에멀젼의지질산화억제능을확인하였다 (Table 4). 1% 오일에멀젼을제조한후해조류추출물을 1,000 ppm을첨가하여 50 C에서저장하면서지질산화억제를확인하였다. 김및다시마추출물이저장기간에따라 3일차에도지질산화를억제하는능력이우수한것으로확인되었다. 다시마추출물의경우라디칼소거능에서는비교적낮은활성을나타냈으나지질산화억제능은가장우수한것으로확인되었다. Park 등 (23) 은다시마분말을첨가한매작과의 TBA값이전반적으로대조군에비해낮은값을나타내지방산화를지연시켰다고보고하였다. Kim 등 (19) 은지질과산화억제효과는미역, 다시마의농도증가에비례하여유의적으로활성이증가되었으며 10 mg/ml 농도에서미역추출물의분무및동결건조물은 61.14 및 50.24% 의저해활성을나타내며해조추출분말의지질산패억제활성은라디칼소거활성에비해높은활성도를나타냈다고보고하였다. 해조류추출물의 tyrosinase 억제활성해조류추출물의 tyrosinase 억제활성을확인하였으며그결과는 Fig. 1과같다. 김, 미역, 다시마및톳추출물의미백활성은각각 25.93, 26.32, 24.76 및 20.24% 의활성을나타냈으며유의적인차이는나타내지않는것으로확인하였다. Choi 등 (24) 등은제주도및남해연안에서해조류를채취한후메탄올로추출한추출물의 tyrosinase 억제활성을분석한결과잎파래, 톳, 미생이및참불가시리등이미약한활성을보였으며, 이외의해조류들은큰활성을보여주지못했다고보고하였다. Kim 등 (25) 은폐미역으로부터조다 Tyrosinase inhibition effect (%). 30 25 20 15 10 5 Fig. 1. Tyrosinase inhibition effect of sea algae extracts powders. 당을추출한후 tyrosinase 억제활성을분석한결과 1, 5, 10 mg/ml 농도증가에따라 0.42, 5.11, 8.06% 의억제활성을확인하였다고보고하였다. 또한해조류를이용한기능성에관한연구로서다시마발효액은 xanthine oxidase를농도에따라유의적으로억제했다고보고하였다 (26). Lee 등 (27) 은가죽나무물추출물의 tyrosinase 저해활성을측정한결과잎부위가 9.31~16.33% 로가장높게나타났으며대조군으로 ascorbic acid(0.1 mg/ml) 는 96.46% 의 tyrosinase 저해활성을나타내는것으로확인하였다. 해조류추출물의총페놀화합물, DPPH 라디칼소거능, ABTS 라디칼소거능및미백활성에대한상관분석을 P< 0.0001 수준에서분석하였으며그결과는 Table 5와같다. 해조류추출물의총페놀함량은 DPPH 및 ABTS 라디칼소거능의결과값과 P<0.0001 수준에서유의적으로상관관계를나타내는것으로확인되었다. 그러나총페놀함량, DPPH 라디칼소거능및 ABTS 라디칼소거능의경우 tyrosinase 억제활성과유의적으로유사하지않는결과를나타내었다. Kwak 등 (28) 은해조류의총플라보이드함량은항산화활성과유의한상관관계가없었으나총폴리페놀함량은지질과산화율, DPPH 라디칼소거율, MDA-BSA conjugation 억제율과유의한양의상관관계를나타냈다고보고하였다. Lee 등 (29) 은보리추출물의항산화및생리활성을분석하여분석치들간의상관관계를분석한결과추출물의총페놀함량과 ABTS 라디칼소거능의경우양의상관관계 (r=0.905, P<0.0001) 를나타냈으며, tyrosinase 억제활성및 ABTS 라디칼소거능의경우도유의적인상관성을나타냈다고보고하였다 (r=0.82, P<0.0001). 본연구결과 Table 5. Correlation coefficients between assays 1) DPPH ABTS TPC TYRO DPPH ABTS TPC TYRO 1 0.979 * 1 0.952 * 0.875 * 1-0.572 ** -0.595-0.447 1 1) DPPH, DPPH radical scavenging activity; ABTS, ABTS radical scavenging activity; TPC, total phenolic compound; TYRO, tyrosinase inhibition activity. * P<0.0001, ** P=1.
해조류추출물의생리활성 1897 해조류추출물의항산화활성및 tyrosinase 억제활성은톳추출물이가장우수하며, 지질산화억제활성의경우김및다시마추출물이높은활성을나타내는것으로확인되었다. 또한해조류추출물의 tyrosinase 억제활성의경우 20.24~26.32% 의억제활성을나타냈으나유의적으로차이를나타내지않았다. 요 우리나라에서소비량인높은대표적인해조류인김, 미역, 다시마및톳을이용하여추출물을제조하여추출물의추출수율, 색도, 총페놀함량, DPPH 라디칼소거능및 ABTS 라디칼소거능을분석하였고, 추출물의지질산화억제능및 tyrosinase 억제활성을분석하였다. 용매를이용하여김, 미역다시마및톳의추출물수율은 3.00, 5.38, 11.73 및 0.72% 를나타내었고, 색도분석결과김, 미역, 다시마및톳추출물의명도는 82.88, 78.53, 83.04 및 78.11로다시마추출물의명도가가장밝은것으로나타났으나김추출물의명도와유의적차이는나타나지않았다. 총페놀함량은김, 미역, 다시마및톳추출물이각각 42.23, 11.59, 10.09 및 46.59 mg/g으로나타냈다. DPPH 라디칼의경우톳추출물이 82.43% 로가장높은라디칼소거능을나타냈으며김추출물의경우 26.76% 의라디칼소거능을확인하였다. ABTS 라디칼소거능의경우 DPPH 라디칼소거능결과와유사한경향을나타내었다. 또한해조류추출물의 tyrosinase inhibition effect 결과는김, 미역, 다시마및톳추출물의 tyrosinase inhibition이각각 25.93, 26.32, 24.76 및 20.24% 의활성을나타냈으며유의적인차이는나타내지않는것으로확인하였다. 해조류추출물의총페놀화합물, DPPH 라디칼소거능, ABTS 라디칼소거능및미백활성에대한상관분석결과해조류추출물의총페놀함량은 DPPH 및 ABTS 라디칼소거능의결과값과 P<0.001 수준에서유의적으로상관관계를나타내는것으로확인되었다. 약 감사의글 이논문은 2011학년도군산대학교신임교수연구비지원에의하여연구되었습니다. REFERENCES 1. Lee SY, Ahn JW, Hwang HJ, Lee SB. 2011. Seaweed biomass resources in Korea. Korean Society for Biotechnology and Bioengineering Journal 26: 267-276. 2. Ahn SM, Hong YK, Kwon GS, Sohn HY. 2010. Evaluation on in-vitro anticoagulation activity of 35 different seaweed extracts. J Life Sci 20: 1640-1647. 3. Jeon YE, Yin XF, Lim SS, Chung CK, Kang IJ. 2012. Antioxidant activities and acetylcholinesterase inhibitory activities from seaweed extracts. J Korean Soc Food Sci Nutr 41: 443-449. 4. Lee HY. 2013. Approval of functional ingredient of health/ functional foods in Korea. Food Industry and Nutrition 18(1): 1-7. 5. Kim SA, Kim J, Woo MK, Kwak CS, Lee MS. 2005. Antimutagenic and cytotoxic effects of ethanol extracts from five kinds of seaweeds. J Korean Soc Food Sci Nutr 34: 451-459. 6. Ebihara K, Kiriyama S. 1990. Physico-chemical property and physiological function of dietary fiber. Nippon Shokuhin Kogyo Gakkaishi 37: 916-933. 7. Kim HS, Kim GJ. 1998. Effects of the feeding Hijikia fusiforme (Harvey) Okamura on lipid composition of serum in dietary hyperlipidemic rats. J Korean Soc Food Sci Nutr 27: 718-723. 8. Jung BM, Ahn CB, Kang SJ, Park JH, Chung DH. 2001. Effects on Hijikia fusiforme extracts on lipid metabolism and liver antioxidative enzyme activities in triton-induced hyperlipidemic rats. J Koran Soc Food Sci Nutr 30: 1184-1189. 9. Kim YY, Lee KW, Kim GB, Cho YJ. 2000. Studies on physiochemical and biological properties of depolymerized alginate from sea tangle, Laminaria japonicus by heating hydrolysis. J Korea Fish Soc 33: 393-398. 10. Lee JS, Lee MH, Koo JG. 2010. Effects of porphyran and insoluble dietary fiber isolated from laver, Porphyra yezoensis, on lipid metabolism in rats fed high fat diet. Korean J Food & Nutr 23: 562-569. 11. Yamamoto L, Nagumo T, Takahashi M, Fujihara M, Suzuki Y, Iizima N. 1981. Antitumor effect of seaweeds. Ⅲ. Antitumor effect of an extract from Sargassum kjellmanianum. Jap J Exp Med 51: 187-189. 12. Cho KJ, Lee YS, Ryu BH. 1990. Antitumor effect and immunology activity of seaweeds toward sarcoma-180. Bull Korean Fish Soc 23: 345-352. 13. Yu L, Haley S, Perret J, Harris M. 2004. Comparison of wheat flours grown at different locations for their antioxidant properties. Food Chem 86: 11-16. 14. Blois MS. 1958. Antioxidant determinations by the use of a stable free radical. Nature 181: 1199-1200. 15. Zhao H, Dong J, Lu J, Chin J, Li Y, Shan L, Lin Y, Fan W, Gu G. 2006. Effects of extraction solvent mixtures on antioxidant activity evaluation and their extraction capacity and selectivity for free phenolic compounds in barley (Hordeum vulgare L.). J Agric Food Chem 54: 7277-7286. 16. Ahn DU, Sell JL, Jeffery M, Jo C, Chen X, WU C, Lee JI. 1997. Dietary vitamin E affects lipid oxidation and total volatiles of irradiated raw turkey meat. J Food Sci 62: 954-658. 17. Jo C, Son JH, Shin MG, Byun MW. 2003. Irradiation effects on color and functional properties of persimmon (Diospyros kaki L. folium) leaf extract and licorice (Glycyrrhiza uralensis Fischer) root extract during storage. Radiat Phys Chem 67: 143-148. 18. Hallagan JB, Allen DC, Borzelleca JF. 1995. The safety and regulatory status of food, drug and cosmetics colour additives exempt from certification. Food Chem Toxic 33: 515-528. 19. Kim JW, Kwon YR, Youn KS. 2012. Quality characteristics and antioxidant properties in spray-dried and freeze-dried powder prepared with powdered seaweed extracts. Korean J Food Sci Technol 44: 716-721. 20. Kim MR, Choir MA, Jeong JS. 2008. Development of fer-
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