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1 J Korean Soc Food Sci Nutr 한국식품영양과학회지 45(12), 1740~1746(2016) 산양삼의품질특성및항산화활성에미치는영향 강경명 1 이진영 1 김명욱 2 이신호 3 1 ( 주 ) 위드네이처 2 경북해양바이오산업연구원 3 대구가톨릭대학교식품공학과 Effects of Quality Characteristics and Antioxidant Activity of Korean Cultivated Wild Ginseng Extract KyoungMyoung Kang 1, JinYoung Lee 1, MyungUk Kim 2, and ShinHo Lee 3 1 R&D Center, WITHNATURE Co., Ltd. 2 Gyeongbuk Institute for Marine BioIndustry 3 Department of Food Science & Technology, Catholic University of Daegu ABSTRACT In this study, we investigated the nutritional and functional constituents as well as quality characteristics and antioxidant activity of Korean cultivated wild ginseng (KG). The chemical compositions and amino acid content of KG were 7.56% water, 73.01% carbohydrates, 12.58% protein, 1.99% lipids, and 5.54% ash as well as mg/g of amino acids, respectively. The major ginsenoside and minor ginsenoside contents of KG were mg/g and 0.04 mg/g, respectively. The total polyphenol and flavonoid contents of KGE (Korean cultivated wild ginseng with 70% ethanol extract) were 8.93 mg GAE/g and 3.96 mg RHE/g, respectively. KGE also showed higher antioxidant activity than the other extracts (KGW, Korean cultivated wild ginseng with water extract) with regard to DPPH and ABTS radical scavenging activities (57.75% and 70.73%, respectively), nitrite oxide scavenging activity (44.01%), SODlike activity (78.05%), reducing power activity (1.08 OD 700 nm), and ferrous ionchelating activity (65.33%). Additionally, KGE had higher elastase, collagenase, and tyrosinase inhibition activities than KGW. These results suggest that KGE can be used as a bioactive and functional material in the food industry. Key words: Korean cultivated wild ginseng, ginsenoside, antioxidant, functionality, phenolic compounds 서 인간은호흡과정에서노화및질병의원인이되는라디칼등의활성산소를생성하며, 인체는체내산화를방어할수있는다양한항산화물질을생성하여항상성을유지한다. 그러나체내항산화체계는다량의산화물을모두방어할수없으므로항산화제를추가섭취하여체내산화를막는것은노화방지및질병예방에매우중요하다. 기존에는효과가빠른합성항산화제가많이사용되었으나, butylated hydroxytoluene과 butylated hydroxyanisole 등의합성항산화제의발암성, 변이원성등이알려지면서 (1), 식물의 2차대사산물을이용한천연항산화제개발이왕성하게진행되고있다. 특히약용식물의약리효과는항산화효과로설명되기도하며 (2), 약용식물이일반식이과채류보다항산화효 Received 22 September 2016; Accepted 8 November 2016 Corresponding author: ShinHo Lee, Department of Food Science & Technology, Catholic University of Daegu, Gyeongsan, Gyeongbuk 38430, Korea leesh@cu.ac.kr, Phone: 론 과가우수한경향을보이므로 (3), 천연항산화제를개발하기위하여다양한민간 전통약용식물들의항산화효과를분석하는것은매우중요하다. 산양삼 (Korean cultivated wild ginseng, KG) 은오가피나무과 (Araliaceae) 인삼속 (Panax) 식물에속하는다년생반음지성숙근초로서한국, 중국, 일본을비롯한동아시아와러시아를비롯한유럽에서까지뿌리를식용또는약용으로이용하여왔다 (4). 이러한산양삼은천연산삼의종자를산림내에자연방임형태로기른것으로산양산삼혹은산양삼이라고한다. 산양삼의유효성분은구조적특징에따라크게사포닌계와비사포닌계로구분할수있다. 사포닌계의생리활성물질로는 ginsenosides가알려져있으며, 비사포닌계는 polyacetylenes, phenolic compounds, acidic polysaccharides, peptides, alkoxides, amino acids 유도체등이있다. 또한, 기타성분으로는 volatile oil, sugar, starch, pectins, minerals 등이있다 (5). 한편이러한산양삼은항암, 혈압강하, 항산화, 간독성등에대한효능이있는것으로알려졌지만 (68), 아직다양한분야에서의연구가미비한상황이다.

2 산양삼의품질특성및항산화활성에미치는영향 1741 본연구는산양삼의이화학적특성과추출용매를달리하여추출한각각의산양삼추출물의항산화활성을검증함으로써산양삼의이용가치를높이고, 기능성식품소재를개발하는데기초자료로활용하고자하였다. 150 rpm으로 24시간교반시켜 2회추출하였다. 각각의추출물은여과지 (Whatman No. 3, Whatman, Maidstone, UK) 로여과한후회전진공농축기로농축하고, 동결건조후분말화하여사용하였다. 재료및방법재료본연구에사용된산양삼은 2013년경북영주시에위치한소백산하산삼영농조합법인에서제공받아세척한후동결건조 (PVTFD20R, Ilshin Lab., Suwon, Korea) 하여분말화한후 70 C deep freezer에보관하면서사용하였다. 일반성분분석일반성분분석은 AOAC 방법 (9) 에따라행하였다. 즉수분은 105 C 상압건조가열법으로, 조단백질은 Kjeldahl법으로, 조지방은 Soxhlet법으로, 조회분은 550 C 직접건식회화법으로, 탄수화물은시료 100 g 중에서수분, 조회분, 조단백, 조지방함량을감하여얻은양으로표시하였다. 아미노산분석동결건조한산양삼 1 g에 70% 에탄올을 10 ml 첨가하여 24시간동안추출과정을거친후 3,000 rpm에서 20분간원심분리하여상등액을취하였다. 상등액은회전진공농축기 (WB 2000, Heidolph, Schwabach, Germany) 로용매를휘발시킨후아미노산분석용 lithium citrate loading buffer로용해시키고, 0.22 μm membrane filter(millipore Co., Billerica, MA, USA) 로여과하여아미노산자동분석기 (Biochrom 30 amino acid analyzer, Biochrom, Cambridge, UK) 로분석하였다. Ginsenosides 함량측정 In 등 (10) 및 Ando 등 (11) 의수포화부탄올추출법으로조사포닌을추출정량하였으며, ginsenosides 조성및함량은조사포닌추출한것을 MeOH에용해한후이를 0.22 μm membrane filter로여과, Acquity UPLC BEH C 18 column(1.7 μm, mm) 을장착한 UPLC(Acquity UPLC System; Waters, Milford, MA, USA) 와이동상용매 A(0.1 % formic acid) 와용매 B(acetonitrile) 를유속 0.3 ml/min 의조건으로시료 10 μl씩주입하여분석하였다. 추출물의제조산양삼의추출방법은시료 100 g에 10배의증류수 (W) 를가한후환류냉각추출과분쇄시료 100 g의 10배의 70% 에탄올 (E) 을가한후상온교반추출로추출물을제조하였다. 환류냉각추출은분쇄시료와증류수를넣은용기에냉각관을부착하여 80 C의항온수조에서 3시간동안추출하였고, 상온교반추출은 25±2 C의실온에서교반기를이용하여 총폴리페놀및플라보노이드함량측정 Singleton 등 (12) 의방법에따라각추출물 1 ml에 0.2 N FolinCiocalteu reagent 1 ml를가하여실온에서 3분간반응시킨후, Na 2CO 3(75 g/l) 1 ml를가한다음암소에서 1시간동안방치한후분광광도계 (Ultrospec 1000, Pharmacia Biotech, Cambridge, UK) 를사용하여 765 nm에서흡광도를측정하였다. 총폴리페놀함량은 gallic acid를표준물질로한표준곡선에의하여산출하였다. 총플라보노이드함량은 Saleh와 Hameed(13) 의방법에따라각추출물 0.1 ml에 5% sodium nitrite 0.15 ml를가한후 25 C에서 6분간방치한다음 10% aluminium chloride 0.3 ml를가하여 25 C에서 5분간방치하였다. 다음 1 N NaOH 1 ml를가하고 vortex 상에서가한후 510 nm에서흡광도를측정하였으며 rutin hydrate(sigmaaldrich Co., St. Louis, MO, USA) 의검량선에의하여함량을산출하였다. DPPH 및 ABTS 라디칼소거능 Blois(14) 의방법을변형하여각추출물 0.4 ml에 0.4 mm DPPH(α,αdiphenylβpicrylhydrazyl) 에탄올용액 0.8 ml를진탕혼합하고 10분간방치후 525 nm에서흡광도를측정하였으며, DPPH radical scavenging ability (%)= 100[(OD of sample/ OD of control) 100] 에의하여활성을산출하였다. ABTS 라디칼소거능은 Re 등 (15) 의방법에따라 7.4 mm ABTS[2,2'azinobis(3ethylbenzothiazoline6 sulfonic acid) diammonium salt] 와 2.6 mm potassium persulfate를혼합하여실온 암소에서 24시간동안방치하여라디칼을형성시킨다음실험직전에 ABTS 용액을 732 nm에서흡광도가 0.700±0.030이되도록 phosphate buffer saline(pbs, ph 7.4) 으로희석하여사용하였다. 희석된용액 950 μl에각추출물 50 μl를가하여암소에서 10분간반응시킨후 732 nm에서흡광도를측정하였으며, ABTS radical scavenging ability (%)=100[(OD of sample/ OD of control) 100] 에의하여활성을산출하였다. 아질산염소거능측정 Kato 등 (16) 의방법에따라각추출물 1 ml에 1 mm NaNO 2 용액 1 ml를가하고 0.1 N HCl을가하여총부피를 10 ml로하였다. 이용액을 37 C에서 1시간반응시킨후 1 ml를취하여 2% 초산용액 4 ml와 30% 초산용액으로용해한 Griess reagent(1% sulfanilic acid : 1% naphthylamine=1:1) 0.4 ml를가한후실온에서 15분간방치하여 520 nm에서흡광도를측정하였으며, nitrite scavenging

3 1742 강경명 이진영 김명욱 이신호 =100[(OD of sample/ OD of control) 100] 에의하여산출하였다. Superoxide dismutase(sod) 유사활성측정 Marklund와 Marklund(17) 의방법에따라각추출물 200 μl에 ph 8.5로조정한 trishcl buffer 용액 3 ml와 7.2 mm pyrogallol 200 μl를가하고 25 C에서 10분간반응시킨후 1 N HCl 1 ml를가하여반응을정지시키고 420 nm 에서흡광도를측정하였으며, SODlike =100 [(OD of sample/ OD of control) 100] 에의하여활성을산출하였다. 환원력측정 Oyaizu(18) 의방법에따라각추출물 2.5 ml에 0.2 M sodium phosphate buffer(ph 6.6) 2.5 ml와 1% potassium ferricyanide[k 3Fe(CN) 6] 2.5 ml를각각혼합하고혼합물을 50 C 항온수조에서 20분간반응시킨다음 10% trichloroacetic acid(tca, CCl 3COOH; w/v) 2.5 ml를첨가하여반응액을 3,000 rpm에서 10분간원심분리한다음상등액 5 ml에증류수 5 ml를첨가하고 700 nm에서흡광도를측정하였다. Ferrous ion chelating 효과측정 Yen 등 (19) 의방법에따라시액 1 ml, 80% 에탄올 0.8 ml, 2 mm FeCl 2 4H 2O[iron(Ⅱ) chloride tetrahydrate] 용액 0.1 ml, 5 mm ferrozine[3(2pyridyl)5,6diphenyl1,2,4triazine4,4 disulfonic acid] 용액 0.1 ml를첨가하여혼합하고실온에서 10분간반응시킨후 562 nm에서흡광도를측정하였다. 계산식 ferrous ion chelating =100[(OD of sample/ OD of control) 100] 에의하여산출하였다. Elastase 저해활성측정 Elastase 저해능측정은 Kraunsoe 등 (20) 의방법에따라 1.0 ml의 Tris/HCl buffer 용액 (0.2 M, ph 8.0) 에 0.1 ml 의 Nsuccinyl(Ala)3pnitroaniline(10.4 mm) 및 1 mg/ml 농도의추출물 0.1 ml를가한후 25 C에서 5분간반응시킨후, elastase( 4.0 units/mg protein, 1 μg/ml) 0.1 ml를가하여 405 nm에서흡광도를측정한다음다시 25 C에서 20분간반응한후의흡광도를측정하였다. 대조군은시료대신 buffer 용액을사용하였다. 저해활성계산식은 elastase inhibitory =100[(OD of sample/ OD of control) 100] 에의하여나타내었다. Collagenase 저해활성측정피부노화억제효과를확인하기위하여 collagenase 저해활성측정은 Wünsch와 Heidrich(21) 의방법으로측정하였다. 즉반응구는 0.1 M TrisHCl buffer(ph 7.5) 에 4 mm CaCl 2 를첨가하여 4phenylazobenzyloxycarbonylPro LeuGlyProArg(0.3 mg/ml) 를녹인기질액 0.25 ml 및시료용액 0.1 ml의혼합액에 collagenase(0.2 mg/ml) 0.15 ml를첨가하여실온에서 20분간방치한후 6% citric acid 0.5 ml를넣어반응을정지시킨다음, ethylacetate 1.5 ml를첨가하여상등액을취한후 320 nm에서흡광도를측정하였으며, collagenase inhibitory = 100[(OD of sample/ OD of control) 100] 에의하여나타내었다. Tyrosinase 저해활성측정 Tyrosinase 저해활성측정은 Vanni 등 (22) 의방법에따라측정하였다. 즉 96well plate에 140 μl의 sodium phosphate buffer(0.05 mm, ph 6.8) 와추출물 100 μl와 40 μl의 Ltyrosine solution(1.5 mm) 을넣고 20 μl의 mushroom tyrosinase(1,500 U/mL) 를넣었다. 반응이잘일어나도록 37 C에서 15분간반응시킨후 492 nm에서흡광도를측정하였으며, tyrosinase inhibitory = 100[(OD of sample/ OD of control) 100] 에의하여나타내었다. 통계처리아미노산, ginsenoside 분석을제외한모든실험은 3회반복으로시행하였으며, 평균치간의유의성은 SPSS system(statistical Package for Social Sciences, SPSS Inc., Chicago, IL, USA) software package(version 12.0) 를이용, P<0.05 수준으로 Duncan's multiple range test에의하여검정하였다. Table 1. Proximate composition of Korean cultivated wild ginseng Composition Percent (%) Moisture Carbohydrate Crude protein Crude lipid Crude ash 결과및고찰 일반성분및아미노산측정산양삼의일반성분을분석한결과는 Table 1과같다. 산양삼의수분함량, 탄수화물, 단백질, 지질, 회분의함량은각각 7.56%, 73.01%, 12.58%, 1.99%, 5.54% 를나타내었으며, 그중탄수화물을가장많이함유하는것으로나타났다. 이러한결과는일반인삼의수분함량, 탄수화물, 단백질, 지질, 회분의함량이각각 8.03%, 62.00%, 12.87%, 2.88%, 4.01% 라는 Lee 등 (23) 의보고와유사한결과를나타내었 7.56±0.08 1) 73.01± ± ± ±0.14 1) Values are means±standard deviation of triplicate determinations.

4 산양삼의품질특성및항산화활성에미치는영향 1743 Table 2. Amino acid composition of Korean cultivated wild ginseng Amino acid Isoleucine * Leucine * Lysine *# Methionine *# Phenylalanine * Threonine * Tryptophan * Valine * Histidine *# Alanine Asparagine Aspartic acid Arginine Glutamic acid Glutamine Glycine Proline Serine Taurine Tyrosine # γaminobutyric acid Content (mg/100 g, dry basis) Total amino acids * Essential amino acids. # Amino acids acting as an antioxidant. 다. 산양삼의아미노산측정결과는 Table 2와같다. 산양삼의총아미노산함량은 mg/100 g을나타내었고, 그중 arginine(10.42 mg/100 g, 64.44%), asparagine(1.25 mg /100 g, 7.73%), γaminobutyric acid(0.87 mg/100 g, 5.38%), glutamine(0.71 mg/100 g, 4.39%), alanine(0.58 mg/100 g, 3.59%), aspartic acid(0.32 mg/100 g, 1.98%) 순으로높은함량을나타내었다. 필수아미노산함량은 1.42 mg/100 g을나타내었고, 그중 leucine이 0.26 mg/100 g으로가장높은함량을나타내었다. 항산화활성이있는아미노산 (24) 으로알려진 lysine, methionine, histidine tyrosine의총함량은 0.62 mg/100 g을나타내었고, 그중 methionine이 0.20 mg/100 g으로가장높은함량을나타내었다. Choi과 Oh(25) 의연구에의하면한국산인삼의아미노산은 arginine> lysine> alanine> aspartic acid> serine 순이었다고보고하였고, Ko 등 (26) 은인삼의아미노산가운데 arginine이가장많았다고보고하였는데, 본연구와다소차이가있는것으로나타났다. 이러한결과는시료들의채취시기, 자연환경, 연령등에많이좌우되기때문에같은 Panax ginseng C.A Meyer 종이라도서로다른결과가나 타날수있다고판단된다. Ginsenosides 함량산양삼의 ginsenoside 조성및함량을분석한결과는 Table 3과같다. 총 ginsenoside 함량은 mg/g을나타내었고, 그중 major ginsenoside 함량은 mg/g으로약 90% 이상의함량을나타내었고, minor ginsenoside 함량은 0.04 mg/g 함량을나타내었다. Major ginsenoside 는 Rb 1(4.68 mg/g)> Re(4.32 mg/g)> Rg 1(2.59 mg/g)> Rc (1.56 mg/g)> Rb 2(1.22 mg/g)> Rf(1.02 mg/g)> Rd(0.35 mg/g)> Rb 3(0.18 mg/g) 순으로높은함량을나타내었고, minor ginsenoside는 Rh 1(0.03 mg/g) 과 Rg 3(0.01 mg/g) 는검출되었지만, Rh 2 와 compound K는검출되지않았다. 인삼은재배조건에따라 ginsenoside 조성및함량에차이가나타나지만대부분 Rg 1, Rb 1, Rb 2, Rc 및 Re를주로함유하고있다는보고 (27) 와유사한결과를나타내었다. 또한, Chang(28) 은인삼의채집시기에따른사포닌성분의함량을분석한결과 Re, Rd, Rg 1 등이전체사포닌의 70% 이상을차지한다고보고한것등과같은결과를나타내었다. 총폴리페놀및플라보노이드함량의변화식물성식품속에함유되어있는많은생리활성물질중페놀은가장많이함유되어있으며, 또한높은항산화활성을가지는것으로알려져있다. 플라보노이드는식물에의해합성된폴리페놀의가장큰부류이며, 효과적인 free radical scavenger로서항산화효과를가진다 (29). 증류수를이용한산양삼추출물 (KGW, Korean cultivated wild ginseng water extract) 과 70% 에탄올을이용한산양삼추출물 (KGE, Korean cultivated wild ginseng ethanol extract) 의총폴리페놀함량을측정한결과는 Table 4와같다. KGW와 KGE의총폴리페놀함량은각각 2.80 mg GAE/g, 8.93 mg GAE/g을나타내었고, 열수추출보다 70% 에탄올추출이더높은함량을나타내었다. 총플라보노이드함량을측정한결과역시 KGW(1.99 mg RHE/g) 보다 KGE(3.96 mg RHE/g) 가더높은함량을나타내었다. 이는추출방법에따라총폴리페놀및총플라보노이드함량에차이가있다는보고 (30) 와유사한경향을나타내었다. 따라서더욱많은유효성분을추출하기위해서는시료에따른최적의추출방법을사용하는것이중요하다고생각한다. DPPH 및 ABTS 라디칼소거능의변화자유라디칼소거능은활성라디칼에전자를공여하여지방질산화를억제시키는척도로사용되고있을뿐아니라 Table 3. Ginsenoside content of Korean cultivated wild ginseng (unit: mg/g) Ginsenosides Total Rb 1 Rb 2 Rb 3 Rc Rd Re Rf Rg 1 Rg 3 Rh 1 Rh 2 CK Korean cultivated wild ginseng

5 1744 강경명 이진영 김명욱 이신호 Table 4. Comparison of total polyphenol and flavonoid content of Korean cultivated wild ginseng water and ethanol extracts Sample 1) KGW KGE Total polyphenol content (mg GAE 2) /g) Flavonoid content (mg RHE 3) /g) 2.80±0.15 b4)5) 1.99±0.02 b 8.93±1.62 a 3.96±0.19 a 1) KGW: Korean cultivated wild ginseng water extract, KGE: Korean cultivated wild ginseng ethanol extract. 2) GAE: Gallic acid equivalent. 3) RHE: Rutin hydrate equivalent. 4) Values are means±sd of triplicate determinations. 5) Means with different letters within each column indicate significant differences (P<0.05). 인체내에서활성라디칼에의한노화를억제하는작용의척도로도이용되고있다 (31). 산양삼추출물의 DPPH와 ABTS 라디칼소거능의측정결과는 Table 5에나타내었다. 1 mg/ml의농도로조정한 KGW와 KGE의 DPPH 라디칼소거능은각각 40.39%, 57.57% 를나타내었고, KGW보다 KGE가더높은항산화활성을나타내었다. ABTS는양이온라디칼을, DPPH는음이온라디칼을소거하는활성을흡광도로측정하는방법으로두방법에대한기질과반응물질과의결합정도가달라추출물을이용한라디칼소거활성측정값에서차이가나타날수있다 (32). ABTS 라디칼소거능은 DPPH 라디칼소거능과유사한결과를나타내었다 (Table 5). 1 mg/ml에서 KGE(70.73%) 가 KGW(51.21%) 에비하여유의적으로높은활성을나타내었다 (P<0.05). 아질산염소거능및 SOD 유사활성의변화추출방법을달리한각각의산양삼추출물 1 mg/ml 농도의아질산염소거능측정결과는 Table 5와같다. 아질산염소거능은 KGW가 23.05%, KGE가 44.12% 로나타났으며, KGE가 KGW보다약 47% 이상높은아질산염분해효과를나타내었다. Kim 등 (33) 의팽이버섯, 하수오, 오미자, 행인등이 20% 이하의소거능을나타내었다는결과와비교하면산양삼은높은아질산염소거능을갖는다고할수있다. 또한, Yamada 등 (34) 은폴리페놀과플라보노이드화합물이종류에따라차이는있으나아질산염을효과적으로분해하여 nitrosamine의생성을억제한다고보고하였는데, 본연 구역시폴리페놀과플라보노이드함량이높은 KGE가높은활성을나타내었다. 산양삼추출물의 SOD 유사활성결과 KGW는 50.16%, KGE는 78.05% 를나타내었다 (Table 5). 이러한결과는약용식물을대상으로한 Lim 등 (35) 의감초 (35.63%), 인진 (25.40%), 황기 (23.13%), 천궁 (18.47%) 등의 SOD 유사활성보다높았으며, 기존에보고된여러종류의천연물보다더높은활성도를갖는다고할수있다. 따라서산양삼추출물은항산화효과가높아다양한기능성소재로활용이가능할것으로판단되었다. 환원력및 ferrous ion chelating 효과의변화각각의산양삼추출물은높은환원력을나타내었고, 그중 KGE는 1.08 OD 700 nm 로가장높은환원력을나타내었다 (Table 5). 환원력은일반적으로페놀함량과상관관계가높다고보고 (36) 되고있으며, 이는본실험결과와도유사하였다. 따라서산양삼에함유되어있는페놀성분이활성산소에수소및전자를공여함으로써활성산소사슬을파괴하여높은환원력을나타내는것으로판단된다. Fe, Cu, Co, Ni, Sn 등과같은산화환원이용이한금속이나이들의금속염은지질산화과정에서촉매로작용할수있는금속이다. 특히일부식품에함유되어있는 Fe 2+ 나 Cu 2+ 등은 hydroxy radical(oh) 과 superoxide radical(o 2) 등의생성을촉진하여식품의지질산화를가속화시키게된다. 이러한금속에대한봉쇄효과는금속촉매제로인한자유라디칼의생성을억제함으로써지질산화를방지할수있는능력을측정하는지표로이용된다 (37). 추출방법을달리한각각의산양삼추출물의 ferrous ion chelating 효과는 KGE(55.33%) 가 KGW(7.14%) 보다높았으며, 약 8배이상의높은항산화활성을나타내었다 (Table 5). Elastase, collagenase 및 tyrosinase 저해활성의변화 Elastin은피부세포외기질 (ECM; extracellular matrix) 을구성하는성분중의하나이며, 피부탄력성저하와주름형성에 collagen의감소뿐아니라피부탄력섬유의구성성분인 elastin 저하도관여한다. Elastase는진피내피부탄력을유지하는데중요한기질단백질인 elastin을분해하는 Table 5. Antioxidant activity of Korean cultivated wild ginseng extracted by different methods Sample 1) KGW KGE AA EDTA DPPH radical scavenging (%) 40.39±2.99 c2)3) 57.57±1.05 b 99.88±0.02 a ABTS radical scavenging (%) 52.21±0.50 c 70.73±3.25 b 99.95±0.02 a Nitrite oxide scavenging (%) 23.05±1.29 c 44.12±4.39 b 99.55±0.28 a Superoxide dismutase like 50.16±5.36 c 78.05±0.77 b 89.81±0.98 a Reducing power (O.D 700 nm) 0.62±0.01 c 1.08±0.01 b 1.25±0.02 a (1 mg/ml) Ferrous ion chelating 7.14±0.42 c 55.33±3.56 b 98.81±0.92 a 1) KGW: Korean cultivated wild ginseng water extract, KGE: Korean cultivated wild ginseng ethanol extract, AA: ascorbic acid (control), EDTA: ethylenediaminetetraacetic acid (control). 2) Values are means±standard deviation of triplicate determinations. 3) Different letters within a column are significantly different (P<0.05).

6 산양삼의품질특성및항산화활성에미치는영향 1745 효소이며, UV 노출로발생하는노화과정과주름형성작용에관여한다. 그러므로 elastase 저해제는피부주름을개선하는작용이있는것으로알려져있다 (38). 이에산양삼추출물의 elastase 저해활성을측정한결과는 Table 6과같다. 각각의산양삼추출물의 elastase 저해활성을측정한결과 KGW 28.99%, KGE 81.96% 로, KGE가가장높은활성을나타내었다. 또한, 대조구로사용된녹차추출물유래의단일물질인 epigallocatechin gallate(egcg) 는 % 를나타내었는데 1 mg/ml의 KGE는 EGCG와유의적이차이는나타났지만, 높은활성을나타내었다. 각각의산양삼추출물의 collagenase 저해활성을측정한결과 KGW는 54.99% 를나타내었고, KGE는 78.96% 로가장높은활성을나타내었다 (Table 6). 특히 KGE는대조구로사용된 mucin(81.81%) 과유의적인차이없이 collagenase를억제하는것으로나타났다. 이러한결과는산양삼추출물 (KGE) 이피부에존재하는 elastin과 collagen의분해를막아피부의주름을개선시킬수있고, 다양한향장제품의기능성소재로써활용이있을것으로판단된다. 일정농도 1 mg/ml에서각각의산양삼추출물의 tyrosinase 저해활성은 KGW 9.99%, KGE 30.96% 를나타내었다 (Table 6). 이러한결과는 Lee 등 (39) 의제주산식물을이용하여 tyrosinase 억제활성을측정한결과 1,000 ppm 의농도에서 10% 의저해효과를나타내었다는결과와비교하면산양삼추출물은높은 tyrosinase 저해능을나타내었다. 이상의결과추출방법을달리한산양삼추출물의항산화물질함량과추출물의항산화효과는각각서로다른경향을나타내었다. 이는식물추출물의항산화효과가추출조건에따라다르게나타난다고보고한 Kim 등 (40) 의결과와같이동일한식물도추출조건을달리함으로써생리활성물질추출수율및생리활성효과를증가시킬수있는것으로판단되었다. 또한, 산양삼추출물은기능성증진을위한다양한식 Table 6. Elastase, collagenase, and tyrosinase inhibition activity of Korean cultivated wild ginseng extracted by different methods (1 mg/ml) Sample 1) KGW KGE EGCG Mucin Kojic acid Elastase inhibition 28.99±6.30 c2)3) 81.96±1.06 b 90.88±1.22 a Collagenase inhibition 54.99±1.30 b 78.96±3.06 a 81.88±1.91 a Tyrosinase inhibition 9.99±5.30 c 30.96±1.06 b 89.88±2.94 a 1) KGW: Korean cultivated wild ginseng water extract, KGE: Korean cultivated wild ginseng ethanol extract, EGCG: epigallocatechin gallate (control), Mucin: control, Kojic acid: control. 2) Values are means±standard deviation of triplicate determinations. 3) Different letters within a column are significantly different (P< 0.05). 품소재로의활용가능성이있을것으로생각하며, 이러한결과가실제로체내에서적용되는지는 in vivo 연구를통하여살펴볼필요가있는것으로생각한다. 요 본연구는산양삼의이용가치를높이고, 기능성식품소재개발을위하여산양삼의이화학적특성과추출용매를달리하여추출한각각의추출물의항산화활성을비교하였다. 산양삼의일반성분은수분 7.56%, 탄수화물 73.01%, 단백질 12.58%, 지질 1.99%, 회분 5.54% 를나타내었고, 총아미노산함량은 mg/100 g이었으며, 그중필수아미노산은 1.42 mg/100 g을나타내었다. 총 ginsenoside 함량은 mg/g을나타내었고, 그중 major ginsenoside(rb 1, Rb 2, Rb 3, Rc, Rd, Re, Rf, Rg 1) 의함량은 mg/g, minor ginsenoside(rg 3, Rh 1, Rh 2) 함량은 0.04 mg/g을나타내었다. 1 mg/ml 농도로조정한증류수를이용한산양삼추출물 (KGW), 70% 에탄올을이용한산양삼추출물 (KGE) 의항산화활성을측정한결과 KGE가모든항목에서가장높게활성을나타냈으며, 각각 8.93 mg/g( 총폴리페놀함량 ), 3.96 mg/g( 총플라보노이드함량 ), 57.57%(DPPH 라디칼소거능 ), 70.73%(ABTS 라디칼소거능 ), 44.12%( 아질산염소거능 ), 78.05%(SOD 유사활성 ), 1.08 O.D 700 nm( 환원력 ), 55.33%(ferrous ion chelating activity) 를나타내었다. 또한, 각각의산양삼추출물의 elastase, collagenase 및 tyrosinase 저해활성을측정한결과역시 KGE가모든항목에서가장높은활성을나타내었으며, 각각 81.96%, 78.96%, 30.96% 를나타내었다. 약 REFERENCES 1. Branen AL Toxicology and biochemistry of butylated hydroxyanisole and butylated hydroxytoluene. J Am Oil Chem Soc 52: Wang TC, Ti MC, Lo SC, Yang CC Free radicalscavenging activity of aqueous extract of Pteris multifida Poiret. Fitoterapia 78: Liu H, Qiu N, Ding H, Yao R Polyphenols contents and antioxidant capacity of 68 Chinese herbals suitable for medical or food uses. Food Res Int 41: Nah SY Ginseng; Recent advances and trends. Korean J Ginseng Sci 21: Nam KY Clinical applications and efficacy of Korean ginseng (Panax ginseng C.A. Meyer). J Ginseng Res 26: Lui JHC, Staba EJ The ginsenosides of various ginseng plants and selected products. J Nat Prod 43: Lee HU, Bae EA, Han MJ, Kim DH Hepatoprotective effect of 20(S)ginsenosides Rg3 and its metabolite 20(S) ginsenoside Rh2 on tertbutyl hydroperoxideinduced liver injury. Biol Pharm Bull 28: Xie JT, Mehendale SR, Li X, Quigg R, Wang X, Wang CZ, Wu JA, Aung HH, Rue PA, Bell GI, Yuan CS Antidiabetic effect of ginsenoside Re in ob/ob mice. Bio

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Journal of Life Science 2011, Vol. 21. No μ μ

Journal of Life Science 2011 Vol. 21. No. 8. 1120~1126 ISSN : 1225-9918 DOI : http://dx.doi.org/10.5352/jls.2011.21.8.1120 μ μ μ α β Journal of Life Science 2011, Vol. 21. No. 8 1121 μ μ 1122 생명과학회지 2011,