• Plant Resources
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• 제6권3호
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• pp.221-226
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• 2003

To improve industrial scale extraction method for extraction of icariin from Epimedium koreanum Nakai, the yields under different extracting conditions such as solvent, temperature, duration and solvent to plant material weight ratio were compared. Regarding extracting solution, highest extracts and icariin yield could be achieved when 10% EtOH was used. In case of plant material to extracting solvent ratio, no significant differences could be observed from 1/10 to 1/50, indicating 1/10 was the most efficient. Extracting temperature significantly affected extracts and icariin yields in that 9$0^{\circ}C$ increased the collected extracts and icariin contents up to 29.6% and 0.76%, respectively, compared to 27.2%, 0.33% at 7$0^{\circ}C$. The yield of extracts was less dependent upon extracting temperature compared to icariin yield. Regarding extraction time, 4 hr and 6 hr resulted in high extracts and icariin yield, respectively. We found extracting Epimedium koreanum Nakai in 10 times volume of 10% EtOH for 4 and 6 hr at 9$0^{\circ}C$ seem to be relatively efficient methods for extracts and icariin, respectively.

• 대한한의학방제학회지
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• 제28권1호
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• pp.81-90
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• 2020

Objectives : This study was conducted to investigate the effects of major constituents of Epimedium koreanum Nakai (Icariin, epimedium A, epimedium B, and epimedium C) on melanin synthesis. Methods : We measured melanin contents, tyrosinase activity, and expression of Rab27a in B16F10 cells cultured with Epimedium koreanum Nakai ethanol extract (EKN) and their major constituents. After treatment with H89 and dibutyryl cAMP, which inhibit or promote the activation of PKA, we observed changes in melanin synthesis and tyrosinase activity stimulated by EKN. Results : Among them, EKN and icariin enhanced tyrosinase activity and melanin contents. We confirmed that EKN augmented melanin synthesis via cAMP/PKA pathway. Icariin-induced tyrosinase activity and melanin content were attenuated by PKA inhibitor H89, while melanogenic effect of icariin was further augmented by cAMP analog, dbc AMP. However, icariin did not affect the expression of small GTPase Rab27a involved in melanosome transport. Conclusions : These results suggest that icariin promotes melanogenesis through cAMP/PKA pathway but does not affect small GTPase Rab27a.

• 한국식생활문화학회지
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• 제32권2호
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• pp.144-149
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• 2017

This study was designed to investigate the contents of icariin during ripening of Epimedium koreanum with sugar and Soju (25, 30, and 35% alcohol contents) using a homemade method. Contents of icariin of Epimedium wine were examined using high performance liquid chromatography. Icariin levels in Epimedium wine increased during the first 3 days of storage; after 6 days, icariin contents decreased gradually until 30 days. Contents of icariin in Epimedium wine by storage temperature (5 and $20^{\circ}C$) were not different during storage. After 30 days, icaiin was not detected in all tested Epimedium wines. Epimedium wines were assessed for 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) and (ABTS) radical scavenging activity. DPPH and ABTS radical scavenging activities increased in early storage periods, however, after 6 or 9 days, activities decreased gradually.

• 한국식생활문화학회지
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• 제33권1호
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• pp.86-94
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• 2018

This study was designed to evaluate the antioxidant activities and protective effects on PC12 cells of the extract of Epimedium koreanum and its main constituents icariin and icariside I. After screening the seven identified flavonoid glycosides from E. koreanum through DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay, E. koreanum, Icariin and Icariside I exhibited significant effect on radical scavenging activity. E. koreanum, icariin and icariside I were examined using DPPH, ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) and FRAP (ferric reducing ability power) assay. In all antioxidant assays, E. koreanum, icariin and icariside I showed high radical scavenging activities in a dose-dependent manner. Protective effects against $H_2O_2-induced$ PC12 cells were assessed with MTT assay. The results indicated that cell viability and protection on PC12 cells of icariside I and icariin increased dose dependently. These study results suggest that E. koreanum, icariin and icariside showed high antioxidant capacities and cell protective effects. Icariside I, one of the metabolites of icariin, may be a new and effective flavonoid compound as a functional component.

• 생약학회지
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• 제32권1호통권124호
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• pp.43-48
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• 2001

Epimedii Herba has been used for tonic, cardiotonic, impotency, amnestic and diuretic. Icariin, a main component was isolated from the Epimedii Herba and identified by the spectroscopic methods. In order to evaluate the quality of Epimedii Herba purchased from various regions of Korea, quantitative determination of icariin in Epimedii Herba using HPLC method has been conducted. Quantitative analysis showed the average amount of icariin in commercial 42 samples was 0.34% on the dry weight basis.

• Journal of Microbiology and Biotechnology
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• 제32권4호
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• pp.437-446
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• 2022

In this study, to obtain icaritin with high pharmacological activities from icariin, which has a content ratio of over 58% in the total flavonoids of Epimedium herb, a special Epimedium flavonoid-glycosidase was produced, purified and characterized from Aspergillus sp.y848 strain. The optimal enzyme production was gained in a medium containing 5% (w/v) wheat bran extract and 0.7% (w/v) Epimedium leaf powder as the enzyme inducer, and strain culture at 30℃ for 6-7 days. The molecular weight of the enzyme was approximately 73.2 kDa; the optimal pH and temperature were 5.0 and 40℃. The enzyme Km and Vmax values for icariin were 15.63 mM and 55.56 mM/h. Moreover, the enzyme hydrolyzed the 7-O-glucosides of icariin into icariside II, and finally hydrolyzed 3-Orhamnoside of icariside II into icaritin. The enzyme also hydrolyzed 7-O-glucosides of epimedin B to sagittatoside B, and then further hydrolyzed terminal 3-O-xyloside of sagittatoside B to icarisiede II, before finally hydrolyzing 3-O-rhamnoside of icarisiede II into icaritin. The enzyme only hydrolyzed 7-O-glucoside of epimedin A or epimedin C into sagittatoside A or sagittatoside C. It is possible to prepare icaritin from the high-content icariin in Epimedium herb using this enzyme. When 2.5% icariin was reacted at 40℃ for 18-20 h by the low-cost crude enzyme, 5.04 g icaritin with 98% purity was obtained from 10 g icariin. Also, the icaritin molar yield was 92.5%. Our results showed icaritin was successfully produced via cost-effective and relatively simple methods from icariin by crude enzyme. Our results should be very useful for the development of medicines from Epimedium herb.

• 생약학회지
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• 제20권1호
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• pp.21-24
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• 1989

A new method for quantitative determination of icariin in Epimedii Herba by high performance liquid chromatography was established. A reversed-phase system with $a\;{\mu}Bondapak\;C_{18}$ column using methanol in water(15% to 70%, gradient elution) as a mobile phase was developed. Icariin and spinosin as an internal reference were detected at 350 nm and the analysis was successfully carried out within 30 min.

• 한국식품영양과학회지
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• 제40권10호
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• pp.1397-1403
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• 2011

본 연구는 삼지구엽초의 기능성 식품학적 이용성 증진을 위한 기초적인 연구로써 우리나라 산야에서 존재하며 예로부터 약용으로 이용되고 있는 삼지구엽초의 주요성분인 icariin을 분리하여 생리활성을 검토하였다. 에탄올로 추출한 삼지구엽초를 극성이 다른 용매인 에틸아세테이트, 부탄올, 헥산, 물, 클로로포름 층으로 분리하여, icariin의 함량을 측정한 결과 각각 27.9, 2.5, 1.7, 1.4, 1.3 ${\mu}g/g$이었고, icariin 수율이 가장 많은 에틸아세테이트 분획물로부터 정제한 icariin 수율은 295.5 ${\mu}g/g$이었으며 대부분 에틸아세테이트에서 용출되는 것을 확인하였다. 용매별 분획물의 항산화효과는 에틸아세테이트 분획층에서 49.0 ${\mu}g/mL$로 가장 높았으며, 부탄올(59.2 ${\mu}g/mL$), 헥산(119.8 ${\mu}g/mL$), 물(122.0 ${\mu}g/mL$), 클로로포름(138.5 ${\mu}g/mL$) 층의 순으로 나타났으며, 정제된 icariin(15.3 ${\mu}g/mL$)은 대조구인 ascorbic acid (19.5 ${\mu}g/mL$), ${\alpha}$-tocopherol(18.2 ${\mu}g/mL$)보다 다소 높은 활성을 나타내었다. Salmonella Typhimurium을 이용한 돌연변이 및 항돌연변이성에 미치는 영향을 조사한 결과, 삼지구엽초에서 분리한 icariin은 TA98과 TA100세포 모두에서 돌연변이성을 나타내지 않았다. 인간면역세포인 B세포와 T세포에서의 면역활성시험을 측정한 결과, icariin은 대조구보다 B세포에서 1.27배, T세포에서 1.28배의 비교적 높은 면역증강활성을 나타내었다. 이러한 결과는 삼지구엽초를 이용한 기능성식품 및 식품재료로서의 이용성 증진을 위한 기초 자료로 제공될 수 있을 것으로 판단된다.

• 한국식품위생안전성학회지
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• 제22권4호
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• pp.359-364
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• 2007

본 연구는 Icariin 함량별로 재배된 삼지구엽초의 추출조건 및 항산화 활성을 비교하기 위하여 수행하였다. 추출 수율은 열수 추출에서는 $90^{\circ}C$에서 7시간, 에탄을 추출에서는 70%와 $50^{\circ}C$에서 3시간 추출한 경우 가장 높은 수율을 나타내었다. 열수추출보다 에탄을 추출이 높은 수율을 나타내었으며 에탄을 추출농도는 100%(추출수율 22.7%)보다는 50%와 70%농도에서 각각 추출수율이 각각 37.3%와 39.3%로 높게 나타났다. 또한, 에탄올 추출시 추출시간과 추출온도 보다는 추출 용매농도가 추출 수율에 더 많은 영향을 미치는 것으로 나타났다. Icariin 함량별 추출수율은 열수 추출과 에탄을 추출 모두 icariin 함량 $0.5{\sim}0.6%$인 대에서 우수하게 나타났으며 icariin 함량에 따른 항산화 활성은 KE9412가 $70^{\circ}C{\cdot}7$시간에서 $34.9({\mu}g/mL),\;90^{\circ}C{\cdot}3$시간에서 $28.4({\mu}g/mL)$로 같은 추출조건에서 KE9405의 $114.3({\mu}g/mL)$ 및 $83.1({\mu}/mL)$에 비하여 각각 3.2배와 2.3배 높은 항산화활성을 나타내었다. 한편, 70% 에탄올 $70^{\circ}C$7시간 추출물로부터 유기 용매별로 순차적으로 분획한 결과, 추출수율은 KE9405와 KE9412 모두 butanol 분획층에서 각각 36.2%와 42%로 가장 높았고 항산화 활성도 butanol 분획층에서 $39.2{\mu}g$으로 가장 높았으며, ethylacetate$(49.0{\mu}g)$, 물$(118.8{\mu}g)$, heaxane$(119.8{\mu}g)$, chloroform$(138.5{\mu}g)$층으로 나타났다.

• 한국식품과학회지
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• 제28권4호
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• pp.616-623
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• 1996

음양곽 성분 중에서 icariin, magnoflorine을 분리하고 이를 표준품으로 음양곽 추출액의 이들 성분에 대하여 HPLC를 이용한 정량법을 수립하였으며 이 성분들의 추출효율을 여러 가지 추출조건하에서 검토하고 그 함량을 분석 하였다. Icariin은 0.1-0.4 mg/ml 농도에서, magnoflorine은 0.02-0.1 mg/ml 농도에서 아래와 같은 조건으로 정량 분석하였다. Icariin의 HPLC조건 :칼럼, LiChrosorbRPl8; 용출용매, MeOH/water/acetic acid (60 : 40 : 0.5); 검출기, 자외부 흡광광도계; 측정파장, 270 nm; 주사량, $25\;{\mu}l$. Icariin의 retention time은 7.37 min으로 다른 성분과도 잘 분리되었다. Icariin 0.1-0.4 mg/ml 50% (v/v)에탄올 용액에 대해 검량선을 작성하여(n=8) 직선을 얻었으며 그 회귀선은 y = -1.4+18296x이고 correlation coefficient (r)=0.9999로 1.00에 매우 접근하였다. Magnoflorine의 HPLC 조건 : 칼럼, LiChrosorb RP18; 용출용매, MeOH/80 mM $Na_{2}\;HPO_{4}/water$ (35 : 35 : 30); 검출기, 자외부 흡광광도계; 측정파장, 270nm ; 주사량, $25\;{\mu}l$. Magnoflorine의 retention time은 11 min이고 $2{\sim}10\;mg/100\;ml$ 50% 에탄올용액을 표준액으로 검량선을 작성하여 직선상의 검량선을 얻었으며(n=6) 그 회귀직선은 y = -0.02+5.7X이고 correlation coefficient (r)=1.0079로 1.00에 근접하였다. 음양곽의 추출용매로 물 또는 50% (v/v)에탄올을 사용하고 추출온도, 추출시간을 달리하였을 때 icariin, magnoflorine, 엑기스의 추출효율을 분석하였다. Icariin은 $50{\%}$ 에탄올($80^{\circ}C$, 1시간)로 추출하였을 때와 물($100^{\circ}C$, 3시간)로 가열 추출하였을 때 거의 같은 정도로 추출되었으나 magnoflorine은 50% 에탄올로 추출하였을 때 더 많이 추출되었다. 반면에 음양곽의 엑기스량은 50% 에탄올로 추출하였을 때 가장 적었고 물로 추출하였을 때는 추출시간이 길면 길수록 더 많았다. 음양곽을 직접 차(茶)로 음용할 때의 추출 조건에서는 icariin 및 magnoflorine의 추출 효율이 약 25%이므로 음양곽 엑기스로 가공한 차제제가 더 바람직하다고 생각된다. 음양곽 중에 icariin및 magnoflorine의 함량은 각각 0.94, 0.16%임을 처음으로 분석하였다.