Kor. J. Hort. Sci. Technol. 28(4):696-700, 2010 순수로즈마리정유제품의생산국별향기성분과항산화활력 우진호ㆍ목민균ㆍ한고운ㆍ이상용ㆍ박권우 * 고려대학교생명공학부 Aroma Components and Antioxidant Activities of Pure Rosemary Essential Oil Goods Produced in Different Countries Jin Ho Woo, Min Gyun Mok, Koh Woon Han, Sang Yong Lee, and Kuen Woo Park * Division of Biotechnology Science, Korea University, Seoul 136-713, Korea Abstract. Thirty-one units of pure rosemary essential oil (EO) in domestic and foreign distribution markets were collected. Aromatic components of these samples were analyzed and antioxidant activity was measured. As the result, major aromatic components were identified such as -pinene, camphene, β-pinene, 1.8-cineole, verbenone and borneol. Major components of rosemary EO were different according to countries. Essential oil from France had good quality. Essential oil cultivated and extracted in South Korea had more content than the collected essential oil in components of α-pinene and camphene. Only 16% of the total 31 unit samples satisfied the content of 9 ingredients presented by ISO. The quality of EDA was shown as 4.8-96.0%, remarkable differences per specimen. A total of 31 units, only 13% of EO showed more than 60% antioxidant activity. Difference of antioxidant activity did not correspond with specific component. Essential oil from Swiss had high antioxidant activity. Additional key words: aromatherapy, camphene, EDA (electron donation ability), pinene 서언 로즈마리는꿀풀과에속하는상록성식물로지중해연안이원산지이며, 주요생산국은프랑스, 스페인, 튀니지, 모로코로연간 300톤의정유가생산된다 (Shrinivas, 2008). 주요구성성분은 pinenes, camphor, camphene, limonene, cineole, borneol, linalool, terpineol, bornyl acetate 등 (Park, 2007) 으로, 간기능활성, 알레르기성비염, 저혈압, 근육통, 두통, 피로해소등의효과 (Kim 등, 2002) 가있는것으로알려져있다. 또한, 강력한항산화효과 (Uchiyanna 등, 1968) 가증명되어현재육제품의산화억제 (Djenane 등, 2003), 김치의발효억제 (Moon 등, 1995) 등각종식품의천연보존제로서이용하기위한연구가활발히진행되고있다. 이처럼이용범위가확대되고있는것에비해유통되고있는정유의성분함량에대한규제와감독은시행되고있지않으며, 기준조차마련되어있지않은실정이다. 그러나로즈마리는한국인이가장선호하는허브 (Park, 2007) 로서대체의학의한분 *Corresponding author: kuenwp@korea.ac.kr Received ***** 20**; Accepted ***** 20**. 야인향기치료 ( 아로마테라피 ) 에서스트레스해소, 기분안정등과같은정신적치료에많이사용되고있다. 따라서본연구에서는국내외에서유통되고있는로즈마리정유를수집하여성분분석과항산화활력을알아봄으로써유통로즈마리정유의국가간품질차이를규명하여소비자의선택및연구응용의기초자료를제공하고천연식품보존제로의이용가능성을규명하고자실시하였다. 재료및방법시험재료순수한 100% 로즈마리정유를 2008년 3월부터 6월까지한국, 일본, 중국, 유럽에서수집한정유 30점과국내 ( 제주도소재제주허브동산 ) 에서재배생산한정유 1점등총 31점을공시분석재료로사용했다. 국가별로는호주산 5점, 프랑스산 7점, 모르코산 3점, 튀니지산 2점, 남아프리카, 독일, 헝가리, 스페인, 스위스, 영국, 미국각 1점그리고원산지미표기 6점이었다 (Fig. 1). 유효유통기간내의정유제품을사용하였다. 696 Kor. J. Hort. Sci. Technol. 28(4), August 2010
Table 1. GC/MSD condition for lavender pure essential oil analysis. GC/MSD control information Oven Initial temp. 50 Initial time 0 min Maximum temp. 325 Final temp. 250 Run time 35 min Fig. 1. Electron donating ability (EDA) content of marked origin in rosemary oils. AFR: South Africa n=1, AUS: Australia n=5, ESP: Spain n=1, FRA: France n=7, GBR: United Kingdom n=1, GER: Germany n=1, HUN: Hungary n=1, KOR: Cultivated in Korea n=1, MAR: Morocco n=3, N/M: Not marked origin n=6, SUI: Swiss n=1, TUN: Tunisia n=2, USA: United States of America n=1. Vertical bars represent means ± standard deviation. 성분측정국내외에서수집한로즈마리정유의방향성분과함량을조사하기위한 gas chromatography(gc) 는 Agilent 6890N (Agilent, USA) 를사용하였으며, mass selective detector(msd) 는 Agilent 5975MSD(Agilent USA) 를각각사용하였다. Column은 Agilent DB-5MS capillary column을사용하였으며, inlet mode는 splitless mode로 flow gas는 helium gas를사용하였다 (Baek, 2006). 세부분석조건은 Table 1과같으며측정시시료는용매 ethyl ether에 10:1의비율로희석한후 auto sampler를사용하여 sample당 1μL씩 3반복주입하였다. 정확한물질동정을위해표준물질 -pinene, 1.8-cineole, borneol(sigma Chemical Co. USA) 을시료와동일하게처리하여측정한후retention time과문헌을비교하였고, NIST mass spectral library version 2.0(2005) program을이용하여측정된 mass spectral data와 mass spectral search program을통해시료와시료반복간의유의성을분석하였다. 동정된로즈마리정유의성분은 ISO(International Standardization Organization, 2009) 에명시한성분기준에따라품질을평가하였다. Inlet Mode Splitless Initial temp. 200 Pressure 16.90 psi Total flow 34.5 ml min -1 Carrier gas Helium Column Type Capillary Model name (number) Agilent DB-5MS (122-5532) Size 30.0 m 250 µm 0.25 µm Detecter (MSD) Acquisition mode Scan Resulting EM voltage 1776.5 였다. 정유의라디컬소거능측정을위해시료 0.2mL과반응액으로조제된 DPPH용액 2.8mL을시험관혼합기에 10초간혼합후상온에서 10분반응시켜 microplate spectrophotometer (Bio-Tek Powerwave XS) 로흡광도를 517nm파장에서측정하였다. 측정된흡광도를전자공여능 (EDA: Electron donating ability) 으로표시하기위해아래와같은식을이용하였다 (KSCS, 2004). EDA(%)=(1-A/B) 100 A : Absorbance of EO sample B : Absorbance of 4 10-4 M DPPH 통계분석시험의변수분석은 SAS program version 9.2(SAS Institute Inc., Cary, NC, USA. 2007) 을이용하여 Duncan의다중검정에의하여처리하였다. 처리반복은독립적으로 3회이상실시하여반복간오차를최소화하였다. DPPH 라디컬소거항산화활력 Blios(1958) 의방법에준하여 4 10-4 M의농도로수용성물질 1,1-diphenyl-2-picryl hydrazyl(dpph, Sigma Chemical Co. USA) 을메탄올에녹여시료의반응액과대조구로사용하 결과및고찰 주요유효성분 ISO의기준주요성분별로함량의분석치를비교한바, Kor. J. Hort. Sci. Technol. 28(4), August 2010 697
ISO에서제시한 9가지성분의기준함량을만족시키는제품은 31점중 5점에불과했다 (Table 2). 이는국내외에서구입하는로즈마리정유의 16% 만이 ISO규격품임을의미한다. 로즈마리의성분중 -pinene은미국산정유 1점에서동정되지않았으며, 호주산제품 1점과영국산제품에서각각 0.9, 1.1% 로현저히낮은함량을나타났다. 그외모든로즈마리정유의 -pinene함량은 9-26% 수준으로 ISO 기준성분함량을모두충족하였으며, 프랑스산정유 1점 (26.3%) 과 국내에서재배하여추출한정유 (25.7%) 에서높은 -pinene 함량이동정되었다 (Table 2). 이는사계절이뚜렷한국내기후가로즈마리의생육환경에영향을주어유효성분중 pinene 함량을높인것으로판단되었다. -pinene은식물계에가장풍부한 terpenoid로서방충, 항균, 생체활성에기여하는물질로알려져있으며, 감기, 감염등의바이러스성질환과신진대사촉진, 혈액순환및지혈등에효능이보고된바있다 (Park, 2008). 따라서국내에서재배된로즈마리정 Table 2. Major aroma components(%) in the ISO standard of domestic and foreign rosemary essential oils. ISO (Min.-Max., %) 9-26 2.5-13 2-9 1-4.5 17-55 0.1-2.5 1-3.5 1-4.5 0-2.5 Brand initial Marked origin -pi. a Cam. b β-pi. c Myr. d 1.8-ci. e Bornyl. f -ter. g Bor. h Ver. i S j EDA (%) HE South Africa 12.57 6.18c 7.22 1.84 32.49 1.42 4.58a 4.34 0.14 9 4.84 J Australia 10.86 5.16 7.90 1.96 26.07-1.08 10.65b - 7 30.21 P Australia 12.83 7.23b 7.12 2.24 24.09 3.27bc 5.30a 7.94-8 66.09bc HM Australia 19.81b x 3.78 5.80 1.45 38.68b 1.34 0.26 6.44-7 13.71 S Australia 0.91 5.83 9.74bc 2.62c 34.20bc 2.58c 0.38 11.13ab - 6 24.14 HI Australia 12.21 4.89 7.08 2.44c 35.40bc 2.05 0.08 9.41bc - 7 29.89 H France 26.25a 4.82 9.86bc - 39.24b 1.22-3.83-6 28.24 ET France 11.88 6.13c 9.25c 2.12 31.31c 2.56c 3.13 5.22-8 5.99 SS France 10.32 5.85 8.38 1.81 36.03bc 2.78c - 10.98b - 7 4.02 WP France 11.27 6.04 8.96 2.37 29.21 2.77-8.84c - 7 16.75 T France 16.80bc 8.47a 1.32 5.92a 20.88 0.69 3.05 6.18 5.34a 8 51.97c PT France 9.45 5.29 7.65-32.94c 0.59 4.05bc 6.31 0.59 8 18.97 AF France 14.16c 6.16 6.21 1.52 29.61 2.16 4.19b 6.20 0.65 9 17.32 FK Germany 11.64 2.96 5.10 1.09 25.11 4.69b 0.07 16.26a - 7 31.61 U Hungary 9.83 5.01 7.62 1.59 29.10 1.11 3.86c 7.69 0.33 9 11.41 JJ Korea 25.73a 8.52a 4.69 3.14bc 23.65 2.59 1.80 5.76 2.97b 9 16.34 AR - z 12.63 6.73bc 5.06 1.79 23.09 6.63a - 8.83c 0.10 8 76.52b M - 11.71 6.14c 7.18 1.97 32.57 0.00 0.16 8.71c 1.49c 7 0.49 PC - 11.66 5.22 6.24 2.10 36.95bc 0.97 5.00a 4.70-8 2.96 Etc - 12.74 5.71 5.90 2.01 35.21bc 0.78 3.31 5.76 0.33 9 3.12 PW - 8.46 2.53 3.79 1.37 57.23a 0.94 1.74 2.62-7 29.31 W - 10.78 0.63 1.54 1.07 65.46a - 3.67c - - 5 9.20 I Morocco 11.39 5.29 8.09 2.34c 26.77 2.68c 0.27 11.10ab - 7 9.69 FP Morocco 9.98 5.56 5.62 2.00 24.88 0.93 0.12 11.96ab 0.62 8 9.03 AG Morocco 10.41 5.37 12.06a - 26.29 2.13 0.23 11.10ab 0.33 7 41.46 SF Spain 14.25c 8.16a 3.67 3.69b 21.71 1.94 0.07 9.70bc 2.99b 8 43.92 E Swiss 10.28 5.89 6.53 2.05 24.20-0.14 0.61 0.30 6 95.98a C Tunisia 10.98 5.43 10.56b - 24.61 3.69bc 0.12 12.24-6 41.95 EX Tunisia 11.17 5.49 8.97 2.40c 27.65 3.27bc 0.37 10.09-7 7.96 B UK 1.14 5.80 8.52 2.03 25.50 4.47b 0.29 14.35-6 74.47bc A USA - y 5.49 7.36 2.79c 36.11bc 1.95 0.22 12.34-6 44.83 z No indication of origin. y No detection. x Mean separation by Duncan s multiple range test at p<0.05. a -pinene, b Capmphene, c β-pinene, d Myrcene, e 1.8-cineole, f Bornyl acetate, g -terpineol, h Borneol, i Verbenone, j Sufficient component count for ISO (9 means fully contained ISO standard). 698 Kor. J. Hort. Sci. Technol. 28(4), August 2010
유의높은 -pinene은의학적이용이기대된다. 국내에서재배하여추출된정유에서는 camphene의함량도다른정유에비하여높게나타났는데, 이는재배지역인제주도의기온의교차가적은해양성기후에따른성분합성의상이한특성으로판단되었다. Camphene은스페인산, β-pinene은튀니지산, 1.8-cineole 은원산지미표기정유에서, camphor는스페인원산과독일원산, borneol은독일원산에서함량이높은것으로평가되었다. 로즈마리의성분중 borneol은독일원산정유 1점이 ISO 기준 4.5% 보다 3.5배이상높은 16.3% 의함량을함유하고있었다 (Table 2). 이는담즙을촉진하는물질로일정량섭취시적당량의담즙분비를촉진하여체내의콜레스테롤수치를낮추고소화를촉진 (Frankel 등, 1996) 하는데기여하는것으로알려져있어, 독일제품이이런효능에따라많이적용할수있을것으로기대되었다. 1.8-cineole은수집된모든정유에서 18.8-65.5% 로 ISO기준에모두충족하였다. 원산지미표기 2점의정유에서각각 65.5, 57.2% 의높은 1.8-cineole 함량을보였다. 로즈마리에함유한 verbenone은호주원산의모든정유에함유되어있지않았으며, 프랑스산정유 1점이 5.3% 로 ISO기준에비해 2배가량많은함량을나타냈다. Terpene류중페르몬의일종인 verbenone성분을많이함유하고있는로즈마리정유는해충을유인하여나무를보호하는보호제 (William 등, 1980) 로의이용도가능할것으로본다. 이는실제로즈마리정유를아로마테라피에이용시성분차이로인한충분한효과를볼수없다는것을의미한다. 그러나 Woo 등 (2010) 은라벤더정유의경우에 ISO규정성분에충족하는정유제품이한점도없다고보고한바있다. 따라서제품의수입이나병 의원에서아로마테라피적용제품의품질이매우중요한것을알수있으며, 이런결과는국내에서처음보고되고있다. 로즈마리정유의 1.8-cineole의경우 ISO기준함량차이가같은지역원산임에도불구하고차이가나는것은재배지역 (Kubeczka와 Formacek, 2002), 재배시기에따른계절적인영향으로판단된다. 결과적으로로즈마리정유의원산지별, 시료별주요유효성분에따른품질은성분별로차이가있고, ISO규정에충족하는제품비율이 16% 로낮은것이확인되었다. 그러므로정유의이용성격과목적에따라성분을고려한선택이이뤄져야할것으로판단되었으며, 무엇보다정유의재배지역, 추출시기및유효성분의함량에대한정확한이력표시가이뤄져서유통되어야이용자의피해를줄이고아로마테라피적용등에효과를증대시킬수있다고판단된다. 항산화활력수집된로즈마리정유 31점의항산화활력을측정한결과, 시료별로 4.8-96.0% 의현저한차이를보였다. 남아프리카원산의정유에서 4.8% 로가장낮은항산화활력을보였으며, 스위스원산의정유가 96.0% 의높은항산화활력을보였고다음이 74.5% 로영국원산제품이었다 (Table 2). 대체로남아프리카의경우, 열대, 아열대기후로인한다양한정유의합성과집적이낮은반면에스위스는밤낮의온도차이가커서정유성분의집적이잘이루어진결과로추측된다 (Park, 2007). 성분분석결과와비교해볼때특정성분에따른항산화활력은일치하지않았으며, 다양한물질에의한항산화활력이나타나는것으로추측되었다. Ryu 등 (2007) 의연구에서로즈마리추출물은 61.3% 의전자공여능 (EDA) 을나타내어우수한항산화활력을인정하였다. 이결과와비슷하거나그이상의항산화활력을보인정유는호주원산정유 (66.1%) 1점, 영국원산정유 (74.5%) 1점, 스위스원산정유 (96.0%) 1점과원산지미표기정유 (76.5%) 1점으로총 4점으로전체 31점중 13% 에불과했었다 (Fig. 1). Yoon과 Kim(2007) 의 polyphenol 함량과항산화활력의관계실험결과를볼때 polyphenol의함량과항산화활력사이에상관관계가있음을밝혔으며, 오렌지의항산화활력을 66.1% 로제시하였다. 이는호주원산정유와수치상으로는같은값이나, 오렌지 poly-phenol 추출물보다정제된로즈마리정유의항산화활력이낮은것으로해석되었다. 또한, 항산화활력의차이는항산화물의농도, 추출방법과추출용매에따라다를수있다 (Lee와 Yoon, 1993). 정유추출온도에따라항산화력과성분함량의차이 (Ibanez 등, 2003) 가있기때문에정유의천연항산화제로서이용을위해서정유추출시항산화성분을잃지않는방법에대한체계적연구와유통기간동안활성물질의휘발과변화를극소화시킬수있는방법을모색하는것이필요할것으로보였다. 그래야만이항산화력이높은정유를골라서식품의보존제로의이용이가능하리라믿기때문이다. 앞으로는음료, 기타가공식품에서정유의사용을통한저장력증진연구를많이실시하여야국내허브정유산업의발전이구축될수있다고판단된다. 또한, 본연구에서유효유통기간내제품을수집하여실험하였으나저장기간중의성분변화가있을것을대비해서생산년월일이유사한제품을선택하여추후비교분석할필요가있다고본다. 초록국내외에서유통되고있는로즈마리정유 31점을수집하여 Kor. J. Hort. Sci. Technol. 28(4), August 2010 699
GC통한유효성분분석과항산화활력시험을실시하였다. 주요성분은 -pinene, camphene, β-pinene, 1.8-cineol, verbenone 및 borneol 등이동정되었다. 주요성분에따라생산국별로함량의차이가나타났으며, 프랑스원산정유의품질이우수하였고, 국내에서재배하여추출한정유가 α-pinene과 camphene의함량이높았다. ISO가제시한 9가지성분의함량을충족하는제품은총 31점중 5점 (16%) 으로낮았다. 전자공여능 (EDA) 은 4.8-96.0% 로현저한차이를보였으며, 60% 이상의항산화능활력을갖는정유는 13% 였다. 특정성분함량에따른항산화활력정도의차이는일치하지않았으며, 스위스원산의정유에서가장높은항산화활력을보였다. 추가주요어 : 아로마테라피, 캄펜, 전자공여능, 피넨 인용문헌 Baek, J.P. 2006. Effect of plant organs, harvest periods, and collected regions on the essential oil components of Lindera obtusiloba Bl. Ph.D Diss., Korea Univ., Korea. Blios, M.S. 1958. Antioxidant determinations by the use of stable free radical. Nature 181:1199-1200. Djenane, D., A.S. Escalante, J.A. Beltran, and P. Roncales. 2003. The shelf-life of beef steak treated with DL-lactic acid and antioxidants and stored under modified atmospheres. J. Food Microbiol. 20:1-7. Frankel, E.N., S.W. Huang, R. Aeschbach, and E. Prior. 1996. Antioxidant activity of a rosemary extract and its constituents, carnosic acid, carnosol, and rosmarinic acid, in bulk oil and oil-in-water emulsion. J. Agric. Food Chem. 44:131-135. Ibanez, E., A. Kubatova, F.J. Senorans, S. Cavero, G. Reglero, and S.B. Hawthorne. 2003. Subcritical water extraction of antioxidant compounds from rosemary plants. J. Agric. Food Chem. 51:375-382. International Organization for Standardization (ISO), 2009. ISO 1342:2000- Oil of rosemary (Rosemarinus officinalis L.). ISO website. http://www.iso.org. Kim, J.C,, M.A. Park, and M.J. Kim. 2002. Aromatherapy in primary care. J. Kor. Acad. Farm Medi. 23:417-429. Korean Society of Crop Science (KSCS). 2004. Analysis and evaluation of the useful component of crop. Hanlimwon. Suwon, Korea. Kubeczka, K.H. and V. Formacek. 2002. Essential oils analysis by capillary gas chromatography and carbon-13 NMR spectroscopy. John Wiley & Sons, Ltd. Second Edition. England. Lee, Y.C. and J.H. Yoon. 1993. Antioxidative effects of volatile oil and oleoresin extracted from rosemary, sage, clove and nutmeg. J. Kor. Food Sci. 25:351-354. Moon, K.D., J.A. Byun, S.J. Kim, and D.S. Han. 1995. Screening of natural preservatives to inhibit Kimchi fermentation. J. Kor. Food Sci. Technol. 27:257-263. Park, K.W. 2007. Herb & Aromatherapy. Sunjinmunhwasa. Seoul. Korea. Park, H.K. 2008. Antimicrobial activities of the essential oils of Rosemarinus offcinalis, Melaleuca altemifolis and Lavendula angustifolia. Thesis for the degree of master. Kyungnam University. Masan, Korea. Ryu, H.Y., K.H. Bae, E.J. Kum, S.J. Park, B.H. Lee, and H.Y. Sohn. 2007. Evaluation for the antimicrobial, antioxidant and antithrombosis activity of natural spices for fresh-cut yam. J. Life Sci. 17:652-657. Shrinivas, P.K. 2008. Market research data on essential oils and absolutes used in fragrance and flavor industry by in business. India. Goarticles Publishing. Uchiyama, M., Y. Suzuki, and K. Fukuzawa. 1968. Biochemical studies of physiological function of tocopheronolactone. Yakugaku Zasshi. 88:678-683. William, D.B., P.E. Tilden, K.Q. Lindahl, D.L. Wood, and P.A. Rauch. 1980. Effects of verbenone and trans-verbenol on the response of Dendroctonus brevicomis to natural and synthetic attractant in the field. J. Chem. Ecology 6:997-1013. Woo, J.H., M.G. Mok, and K.W. Park. 2010. Aroma components and antioxidant activities of pure lavender essential oil goods in different produced countries. Kor. J. Hort. Sci. Technol. 28:138-143. Yoon, K.E. and A.N. Kim. 2007. Total poly-phenol compounds and antioxidant activity in horticultural crops. J. Kor. Soc. People, Plant and Envir. 10:74-83. 700 Kor. J. Hort. Sci. Technol. 28(4), August 2010