• 한국작물학회지
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• 제53권2호
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• pp.224-232
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• 2008

고 기능성 유용 차나무 품종육성 및 녹차의 이용성 증진 연구의 기초 자료를 제공하기 위해 녹차 함유 catechin 화합물 및 caffeine 성분을 동시에 분석할 수 있고, 재현성 및 검출 감도가 증진된 최적의 HPLC 분석 조건을 검토한 결과를 요약하면 다음과 같다. 1. 녹차에 함유된 catechin 화합물 및 caffeine의 동시분석을 위한 최적 HPLC 분석 조건을 검토한 결과 컬럼은 YMC-pak ODS-AM 303(25 cm)컬럼을 이용하고, 검출파장은 280 nm, 컬럼온도를 $30^{\circ}C$, 분당유속은 1 mL, 이동상 용매로는 A용매로 0.1% 인산 - 증류수, B용매로 100% MeOH를 사용하여 농도구배 조건으로 분석하는 것이 가장 효율적 분석법으로 조사되었다. 2. 녹차에 함유된 catechin 화합물 및 caffeine을 동시에 분석할 수 있는 최적 HPLC분석조건의 재현성 검정을 실시한 결과 머무름 시간(Rt.)의 변이계수는 최대 1.23% 미만, peak area의 변이계수는 최대 5.13% 미만을 나타내어 확립된 녹차 함유 catechin 9종 화합물 및 caffeine의 HPLC 정량분석법은 재현성이 높은 분석법으로 확인되었고, 각 cafechin 화합물 및 caffeine 성분의 최소 검출한계는 최소 50ppb(0.050 ppm, 50 ng/mL)이하로 조사되어, 확립된 HPLC 분석법은 녹차의 작물학적 특성 평가용으로 활용하기에 전혀 무리가 없는 분석법으로 판단되었다.

• 한국식품과학회지
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• 제35권3호
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• pp.503-509
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• 2003

비만 저해 물질을 탐색하고자 전통의학에서 사용되는 200여 종류의 식용 또는 약용식물을 대상으로 지방세포 분화저해활성을 보이는 생약식물을 선별하여 용매층으로 활성이 이행되는 생약 중 황백(Phellodendri Cortex)을 최종적으로 후보식물로 선정하였다. 황백을 메탄올로 추출한 후, silica gel column chromatography, ODS RP-18 column chromatography, HPLC 등을 수행하여 지방세포 분화 저해활성을 갖는 화합물 PC-4를 분리하였다. PC-4는 노란색의 분말형태로 메탄올을 용매로 UV 최대흡수치를 조사한 결과, 222, 265, 349, 429 nm에서 최대의 UV의 흡수치를 보였으며 산이나 알카리 조건에서 최대 흡수치의 변화가 관찰되지 않았다. EI-mass spectrum을 측정한 결과, 분자량이 336.1로 예상되었으며 $^1H-NMR,\;^{13}C-NMR,\;DEPT,\;^1H-^1H\;COSY$, HMQC, HMBC NMR spectrum을 통해, PC-4는 isoquionoline alkaloide계열의 berberine으로 동정되었다. PC-4는 지방의 대사에 관여하는 중요한 효소인 fatty acid synthase와 pancreatic lipase에 대한 저해활성은 나타나지 않았으나, 지방세포 분화 저해활성은 $1\;{\mu}g/mL$의 낮은 농도에서도 관찰되었다.

• 대한화학회지
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• 제37권8호
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• pp.711-716
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• 1993

천연 알칼로이드의 하나인 ${\iota}$-sparteine (SP)의 전기화학적 산화반응을 아세토니트릴 용액 중에서 조사하였다. SP의 순환전압전류그림(CV)은 Ag/AgCl (0.1M AgNO$_3$ in acetonitrile) 전극에 대하여 +0.75 V and +1.45 V에서 두 개의 비가역적 산화봉우리를 나타내었다. 두 과정은 각각 약 1.2~1.3개의 전자반응에 해당함을 전기량법을 이용하여 확인하였다. SP 의 첫 단계산화는 SP의 질소원자 하나가 산화되어 SP 라디칼 양이온을 형성하고, 라디칼 양이온은 뒤 이은 빠른 탈수소반응에 의하여 중성의 이민 라디칼을 형성한다. 이 중성 라디칼은 두 가지 반응 경로가 가능하다:대부분의 중성 라디칼은 불균등화 반응에 의하여 SP와 엔아민으로 된다. 또한 일부의 중성 라디칼은 다시 일전자 산화반응에 의해 1,2-dehydrosparteinium 양이온을 생성하게 된다. 이 양이온을 KOH로 처리하면 (+)-lupanine으로 된다. SP를 전기분해하여 얻은 생성물들을 분리한 후 적외선분광법, 질량분석법, 자외선-가시광선 분광법 및 얇은 막 전기분광화학법을 이용하여 확인하였다.

• Molecules and Cells
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• 제38권4호
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• pp.327-335
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• 2015

Piperlongumine, a natural alkaloid isolated from the long pepper, selectively increases reactive oxygen species production and apoptotic cell death in cancer cells but not in normal cells. However, the molecular mechanism underlying piperlongumine-induced selective killing of cancer cells remains unclear. In the present study, we observed that human breast cancer MCF-7 cells are sensitive to piperlongumine-induced apoptosis relative to human MCF-10A breast epithelial cells. Interestingly, this opposing effect of piperlongumine appears to be mediated by heme oxygenase-1 (HO-1). Piperlongumine upregulated HO-1 expression through the activation of nuclear factor-erythroid-2-related factor-2 (Nrf2) signaling in both MCF-7 and MCF-10A cells. However, knockdown of HO-1 expression and pharmacological inhibition of its activity abolished the ability of piperlongumine to induce apoptosis in MCF-7 cells, whereas those promoted apoptosis in MCF-10A cells, indicating that HO-1 has anti-tumor functions in cancer cells but cytoprotective functions in normal cells. Moreover, it was found that piperlongumine-induced Nrf2 activation, HO-1 expression and cancer cell apoptosis are not dependent on the generation of reactive oxygen species. Instead, piperlongumine, which bears electrophilic ${\alpha},{\beta}$-unsaturated carbonyl groups, appears to inactivate Kelch-like ECH-associated protein-1 (Keap1) through thiol modification, thereby activating the Nrf2/HO-1 pathway and subsequently upregulating HO-1 expression, which accounts for piperlongumine-induced apoptosis in cancer cells. Taken together, these findings suggest that direct interaction of piperlongumine with Keap1 leads to the upregulation of Nrf2-mediated HO-1 expression, and HO-1 determines the differential response of breast normal cells and cancer cells to piperlongumine.

• 대한한의학회지
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• 제28권1호통권69호
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• pp.105-116
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• 2007

Objectives : Mahuang (Ephedra sinica), well known as an herbal medicine in the East and West, contains a relatively high percentage of ephedrine known as sympathomimetic alkaloid. We investigated the effects of Mahuang on sympathetic nervous system with heart rate variety (HRV). Time and frequency domain analysis of HRV is a noninvasive technique capable of providing information on autonomic modulation of the sinus node. Methods : We investigated 57 healthy volunteers consisting of 37 subjects in the Mahuang group and 20 subjects in the placebo group. Study form was a randomized, placebo-controlled, double-blind clinical trial. The 37 subjects in the Mahuang group took 3 Mahuang capsules (1 capsule equivalent to 2g herb Mahuang) twice a day at 10 a.m. and 2 p.,., while the 20 subjects in the placebo group took 3 placebo-capsules filled with glutinous rice powder at the same times. Mahuang medicine and placebo medicine were made into opaque capsules. We measured HRV at 3 p.m. 1 or 2 days before medication and at 3 p.m. after medication. Results : Mean-RR and SDNN of the Mahuang group significantly decreased compared with that of the placebo group, but the heart rate of the Mahuang group significantly increased compared with the placebo. HRV-Index, RMSSD and SDSD of the Mahuang group significantly decreased compared with that of the placebo group, but PNN50 of the Mahuang group significantly increased compared with placebo. Ln(TP), Ln(VLF), Ln(LF) and Ln(HF) of the Mahuang group significantly decreased compared with those of the placebo group. There were no significant differences in normalized LF, normalized HF and LF/HF ratio between the Mahuang and placebo groups. Conclusion : The results suggest that Mahuang in healthy adults tends to reduce the autonomic nervous system within the normal range.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.14-14
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• 2018

This Camptotheca acuminata Decne. (CA), belonging to Nyssaceae, is a deciduous tree. and has been used as Traditional Chinese medicine since ancient times. The CA produces camptothecin a natural indole alkaloid, and reported to have anti-cancer effects. But the studies on biological activities of CA leaves are insufficient. Therefore, this study confirmed various biological activities such as antioxidant, antidiabetic, anticancer, antiinflammatory and metabolism analysis by HPLC-MS/MS of CA leaves. The $RC_{50}$ values of DPPH radical scavenging activity of ethyl acetate fraction, n-Butanol fraction, methanol extraction, water fraction and n-Hexane fraction were $12.23{\pm}0.01$, $15.93{\pm}0.42$, $55.12{\pm}0.45$, $56.29{\pm}4.15$ and $427.29{\pm}6.13ug/mL$, respectively. The $IC_{50}$ values of ${\alpha}$-glucosidase inhibitory activity of ethyl acetate fraction, n-Butanol fraction, methanol extraction, n-Hexane fraction and water fraction were $24.29{\pm}0.14$, $47.86{\pm}0.45$, $54.23{\pm}1.21$ $466.76{\pm}2.21$ and $623.91{\pm}9.67ug/mL$, respectively. The nitric oxide release activity of n-Hexane fraction, methanol extraction, ethyl acetate fraction, water fraction and n-Butanol fraction were $31.49{\pm}5.74$, $29.79{\pm}0.71$, $26.89{\pm}0.71$, $8.24{\pm}5.83$ and $7.75{\pm}4.08%$ at 25 ug/mL, respectively. The anti-cancer activity of n-Hexane fraction, methanol extraction, ethyl acetate fraction, water fraction and n-Butanol fraction were $31.49{\pm}5.74$, $29.79{\pm}0.71$, $26.89{\pm}0.71$, $8.24{\pm}5.83$ and $7.75{\pm}4.08%$ at 25 ug/mL, respectively. The ethyl acetate fraction activities showed higher biological activities than other fractions. Thus, Additional studies were conducted using ethyl acetate fraction. Metabolite analysis was performed using a LCMS-8040 triple quadrupole mass spectrometer. As a result, Five compounds (1-5) were identified in the ethyl acetate fraction of the CA leave. The identification of these compounds was generated by the analysis of fragmentation methods of the negative and positive ion modes. Five compounds were identified as gallic acid (1), chlorogenic acid (2), isoquercetin (3), astragalin (4) and camptothecin (5). These results suggest that the CA leave can be used for functional materials.

• KSBB Journal
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• 제24권2호
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• pp.195-200
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• 2009

본 연구에서는 Eschscholtzia californica의 이차 대사산물인 benzo[c]phenanthridine alkaloids의 생산량을 향상시키기 위해 주요 영양성분인 질소원 농도를 조절하여 배지의 조성을 최적화하였다. 그 결과, 질산과 암모늄 이온의 초기 농도 비율은 세포 성장과 alkaloids 생산량 증대에 중요한 영향을 미치는 인자로서 작용함을 볼 수 있었다. 총 질소 농도를 standard MS배지의 총 질소원 농도와 동일하게 유지하고 (60 mM), 농도 비율 다양하게 조절했을 때, 세포 성장과 alkaloids 생산량은 질산과 암모늄 이온을 각각 단독으로 이용할 때 보다 효율적으로 증대함을 볼 수 있었다. 최대 성장 (9.84 g DCW/L)과 alkaloids 생산량 (60.72 mg/L)은 50 : 10의 비율에서 나타났으며, 농도 비율의 감소는 세포 성장을 억제하고 alkaloids 생산량을 감소시켰다. 또한, 질산과 암모늄 이온이 세포 성장과 alkaloids 생산량에 미치는 영향력을 보다 명확히 확인하기 위해 질산 이온과 암모늄 이온 농도를 각각 조절한 결과, 암모늄 이온이 증가할수록 alkaloids 생산량은 비슷하나, 세포 성장은 감소하였으며, 또한, 질산 이온의 농도를 증가시킬수록 세포 성장은 비슷한 값을 나타내었지만, alkaloids 생산량은 다양한 차이를 나타내었다. 본 실험에서 세포성장과 alkaloids 생산량에 적합한 농도는 50 : 25 (mol/mol) 비율에서 나타났으며, 본 실험을 통해 Eschscholtzia californica의 현탁 세포배양에서 질소원 농도의 조절을 통한 외부 환경조절은 세포 성장과 alkaloids 향상에 매우 효과적이며, 소량의 암모늄 이온의 첨가 시 질산 이온 농도조절은 세포 성장을 억제하지 않으면서 alkaloids 생산량을 유도하는데 적합한 이온임을 확인하였다.

• 약학회지
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• 제57권2호
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• pp.87-94
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• 2013

An analytical methodology based on solid-space extraction (SPE) with with Bond Elut Certify cartridge (Varian, 130 mg) has been developed for the qualification and quantitation of strychnine in blood. After the elution layer was evaporated, the residue was reconstituted with methanol for GC/MS. Internal standard was used 10 mg/l dextromethorphan. Strychnine is a potent central nervous stimulant and convulsant, and an alkaloid found in seeds of Strychnos nux-vomica. It was used therapeutically to improve circulation and muscle tone in oral or intramuscular doses of 0.05~8 mg. The fatal dose of strychnine for humans is 50~100 mg. A man was found dead lying curled up the corner of the large room in a roof house after the fire fighter opened a locked door inside to put out the fire. The postmortem blood and gastric contents were analyzed for toxicological testing. Strychnine and brucine were detected using GC/MS first in gastric contents extracts. The contents of strychnine was 0.083 mg/l in heart blood, 0.088 mg/l in peripheral blood and 4.0 mg/kg in gastric contents, respectively. Method validation was carried out in terms of linearity, accuracy, precision (intraday, interday) in blood. The assay is linear over 0.05~10 mg/l ($r^2$=0.999). Limit of detection (LOD) and limit of quantitation (LOQ) in blood were determined 0.02 mg/l (S/N=3) and 0.07 mg/l (S/N=10), respectively. Accuracy (bias%) of strychnine with 0.1, 1 and 10 mg/l was 12.0% (n=6), 9.3% (n=6) and 6.9% (n=6), respectively. Intraday precision (CV%) of strychnine with, 0.1, 1 and 10 mg/l were 6.4%, 10.4%, 1.2% (n=6), respectively. Interday precision (CV%) of strychnine with 0.1, 1 and 10 mg/l over three days were 24.0%, 18.5%, 13.8% (n=18), respectively. Relative recovery with 0.1, 1 and 10 mg/l (in blood) were 114.9%, 99.3% and 87.4% (n=6), respectively. The described method can be applied in forensic toxicology to determine strychnine in blood samples.

• The Korean Journal of Physiology
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• 제15권2호
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• pp.91-96
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• 1981

The wormwood is one of the plants which occur widely throughout the world. Though the precise data on the entire chemical composition of mugwort leaves are not available, the major principles which have been found so far include inulin, alkaloid, thujon, sesquiterpene and several vitamins. Santonin, a parasiticide, is one of the glucosides extracted from the limited species of wormwood. It has long been known in herb medicine that the plants of this family has not only strong hemostatic, analgesic and parasiticidal actions but also therapeutic effects for diarrhea, stomachache and asthma. In recent pharmaceutical botany the wormwood is introduced to have antipyretic and astringent actions also. The mugwort(Artemisia asiatica Nakai) is the most common species of wormwood that occurs in Korea. The usage of this edible leaves of mugwort is rather various. It is used not only for wormwood bath but also as forage, moxa and medicinal agents. Recently Kim et al reported from their study on the effect of mugwort on the motility of isolated intestine of rabbits that tonus and motility were markedly enhanced by mugwort but this effect of mugwort on intestinal motility was almost completely blocked by atropine suggesting that activity of mugwort was exerted through its cholinergic effect. It was the findings of Kim et al that prompted the authors to do the present experiment. The present study was undertaken to investigate effects of mugwort(Artemisia asiatica Nakai) juice on the respiration and blood pressure in cats. And also studied was the mechanism of depressor action of Artemisia asiatica Nakai Juice (AAJ). The results obtained are as follows; 1) It was observed that mean arterial blood pressure and heart rate were decreased markedly by AAJ. Following administration of 0.15 ml/kg and 0.3 ml/kg AAJ into cats the maximum depressor responses observed were $77.5{\pm}2.2\;mmHg$ and $94.0{\pm}3.7\;mmHg$ respectively. 2) Depressor responses to AAJ were blocked markedly by atropine whereas the responses were not affected by propranolol and dibenamine. Therefore it is strongly inferred that depressor action of AAJ results mainly from its cholinergic effect. This inference was further substantiated by the fact that heart rate change which invariably accompanies depressor responses to AAJ was almost completely abolished by atropinization. 3) After administration of AAJ into cats frequency of respiration was markedly increased while depth of respiration decreased during first 2-3 seconds.

• 한국환경성돌연변이발암원학회지
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• 제26권4호
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• pp.119-124
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• 2006

There are $10{\sim}30%$ polyphenol and $2{\sim}4%$ caffeine in green tea. Caffeine is a kind of alkaloid containing nitrogen which cause stimulation, impatience, headache, insomnia, low birth weight infant. Because of these negative effect, decaffeined beverage came out and decaffeined coffee already have a big market since 1970s. Having proving the physiologic functions of green tea, high consumption of coffee is shifting to green tea. Because of the carcinogenic effect of the organic solvents, decaffeine processing with supercritical carbon dioxide has industrialized and have an advantage in environment-friendly and minimized flavor loss. Decaffeined green tea using supercritical carbon dioxide is considered to be safe but there are not enough study. We investigated the chromosome aberration test with mammalian cell line, CHL. When the cells were treated with 5000, 2000, 1000 ${\mu}g/ml$ and compared with the negative controls, there were no significant(P>0.05) increased chromosome aberration. Same results was observed when adding S9 mixture or not. As a result, water extract of decaffeined green tea using supercritical carbon dioxide does not induce chromosome aberration.