• Title/Summary/Keyword: Alkaloid

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Optimal HPLC Condition for Simultaneous Determination of Catechins and Caffeine in Green Tea Extracts (녹차 함유 카테친 및 카페인 동시분석을 위한 최적 HPLC 분석 조건)

  • Choung, Myoung-Gun;Lee, Min-Seuk
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.2
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    • pp.224-232
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    • 2008
  • The health benefits associated with tea consumption have resulted in the wide inclusion of green tea extracts in botanical dietary supplements, which are widely consumed as adjuvants for complementary and alternative medicines. Tea contains polyphenols such as catechins or flavan-3-ols including (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), and (-)-epigallocatechin gallate (EGCG), as well as the alkaloid, caffeine. The contents of catechins and caffeine in green tea are considered as a standard of quality evaluation of green tea. Therefor, the purpose of this study was to investigate the most suitable HPLC condition for simultaneous determination of catechins and caffeine in green tea extracts. The efficient HPLC analytical condition of catechins and caffeine contained green tea extracts was developed. The gradient elution employed a $250\;mm\;{\times}\;4.6\;mm$ i.d. YMC-pak ODS-AM 303 column. The gradient system was used two mobile phases. A gradient elution was performed with mobile phase A, consisting of 0.1% aqueous phosphoric acid, and mobile phase B, comprising 100% MeOH, and delivered at a flow rate of 1 mL/min as follows: $0{\sim}25\;min$, 80% A; $26{\sim}50\;min$, $80{\sim}70%$ A; 51 min, 80% A. $51{\sim}55\;min$, 80% A. The UV detection wavelength was set at 280 nm. The limit of detection (LOD) for catechins and caffeine standards were under 50 ng/mL.

Adipocyte Differentiation Inhibitor Isolated from the Barks of Phellodendron amurense (황백(Phellodendri Cortex)으로부터 분리한 지방세포 분화 저해물질)

  • Kim, Kyung-Hee;Ahn, Soon-Cheol;Lee, Myung-Sun;Kweon, Oh-Song;Oh, Won-Keun;Kim, Min-Soo;Sohn, Cheon-Bae;Ahn, Jong-Seog
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.503-509
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    • 2003
  • For the development of the anti-obesity natural drug, the inhibitor of adipocyte differentiation was screened from Korean traditional medicinal plants. Phellodendri Cortex was selected as a candidate of adipocyte differentiation inhibitor. An inhibitory compound PC-4 was purified from the methanol (MeOH) extract of Phellodendri Cortex using silica gel and ODS RP-18 column chromatography and HPLC. PC-4 was obtained as yellow powder; UV ${\lambda}_{max}$ (MeOH): 230, 260, 340 and 430 nm. The chemical structure of PC-4 was determined as an isoquionoline alkaloid, berberine, on the basis of various NMR experiments including $^1H-\;and\;^{13}C-NMR$. The PC-4 inhibited the differentiation of preadipocyte NIH-3T3 L1 cells at a concentration of $1\;{\mu}g/mL$.

Mechanistic Studies for Electrochemical Oxidation of ${\iota}$-Sparteine (${\iota}$-Sparteine의 전기화학적 산화반응에 대한 메카니즘의 연구)

  • Jin-Hyo Park;Chang-Soo Jin;Sung-Nak Choi;Yoon-Bo Shim
    • Journal of the Korean Chemical Society
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    • v.37 no.8
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    • pp.711-716
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    • 1993
  • The mechanism for electrochemical oxidation of natural alkaloid, ${\iota}$-sparteine (SP) was studied in acetonitrile solvent. The cyclic voltammogram of SP shows two irreversible anodic peaks at +0.75 V and +1.45 V vs. Ag/AgCl (0.1M AgNO$_2$ in acetonitrile) electrode. Coulometry reveals that the number of electrons involved in each oxidation peaks is in the range of 1.2∼1.3 respectively. Neutral imine radical was produced by fast deprotonation of SP radical cation formed by oxidation of one nitrogen atom in SP. Two pathways are possible for the reaction of the neutral radical: Due to the disproportionation of the radical, SP and enamine were mainly produced. Also, the 1,2-dehydrosparteinium cation was formed as minor product through the second one electron transfer oxidation of this radical. The (+)-lupanine was produced by treatment of sparteinium cation with potassium hydroxide. We have isolated and confirmed the electrolysis products using IR, GC-MS, UV-Vis, and thin-layer spectroelectrochemical method.

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Heme Oxygenase-1 Determines the Differential Response of Breast Cancer and Normal Cells to Piperlongumine

  • Lee, Ha-Na;Jin, Hyeon-Ok;Park, Jin-Ah;Kim, Jin-Hee;Kim, Ji-Young;Kim, BoRa;Kim, Wonki;Hong, Sung-Eun;Lee, Yun-Han;Chang, Yoon Hwan;Hong, Seok-Il;Hong, Young Jun;Park, In-Chul;Surh, Young-Joon;Lee, Jin Kyung
    • Molecules and Cells
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    • v.38 no.4
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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.

Effect of Mahuang on Heart Rate Variability in Adults : a Double-Blind, Placebo-Controlled, Randomized Trial (마황 복용이 정산인의 심박변이도에 미치는 영향에 대한 무작위배정 이중맹검 임상연구)

  • Son, Dong-Hyuk;Hsing, Li-Chang;Kim, Lak-Hyung;Jeong, Seung-Il;Seo, Eui-Seok;Jang, In-Soo
    • The Journal of Korean Medicine
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    • v.28 no.1 s.69
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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.

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Biological Activities and the Metabolite Analysis of Camptotheca acuminata Dence.

  • Cho, Jwa Yeong;Park, Mi Jin;Ryu, Da Hye;Kang, Young-Hwa
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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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.

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Improvement of Growth and Benzo[c]phenanthridine Alkaloids Production by Modifying Nitrogen Source in Suspension Cell Culture of Eschscholtzia californica (Eschscholtzia californica의 현탁 세포배양에서 질소원 조절에 의한 세포 성장 및 Benzo[c]phenanthridine Alkaloids 생산량 향상)

  • Lee, Song-Eun;Rhee, Hong-Soon;Son, Seok-Young;Park, Jong-Moon
    • KSBB Journal
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    • v.24 no.2
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    • pp.195-200
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    • 2009
  • The effect of nitrogen source on cell growth and benzo[c]phenanthridine alkaloids production by modifying $NO_3\;^-:NH_4\;^+$ ratio in cell suspension culture of Eschscholtzia califarnica was investigated. When total nitrogen concentration is maintained (60 mM), maximum benzo[c]phenanthridine alkaloids production is about 60.72 mg/L at 50:10 (mol/mol). This productivity was 3.8 times higher than that obtained when cells were grown instandard MS medium. The decrease of $NO_3\;^-:NH_4\;^+$ ratio at 60 mM of total nitrogen caused the decline of both growth and benzo[c]phenanthridine alkaloids production. Under the same concentration of $N0_3\;^-$ (50 mM), higher concentration of $NH_4\;^+$ inhibited cell growth strongly but induced alkaloids production slightly. Also, under the same concentration of $NH_4\;^+$ (25 mM), higher concentration of $N0_3\;^-$ induced alkaloids production strongly but high concentration of $N0_3\;^-$ (${\geq}$100 mM) interfered alkaloids instead. Maximum benzo[c]phenanthridine alkaloids production is about 62.71 mg/L at 50:25 (mol/mol). These results suggest that higher biomass and higher alkaloids production could be obtained by optimizing each nitrogen concentration as well as $NO_3\;^-:NH_4\;^+$ ratio in the culture medium. Nitrate and ammonium in culture medium have distinct role in the regulation of growth and alkaloids production; ammonium had a strong influence on growth while nitrate had an influence on alkaloids production.

Method Development and Validation of Strychnine in Blood by Gas Chromatography/Mass Spectrometry Using Solid Phase Extraction and its Application in Real Specimens (SPE 및 GC/MS에 의한 혈액중 스트리크닌의 분석법 개발 및 검출사례)

  • Rhee, Jongsook;Yum, Hyesun;Moon, Sungmin;Lee, Sangki
    • YAKHAK HOEJI
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    • v.57 no.2
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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.

Depressor Responses to Intravenously Administered Artemisia asiatica Nakai Juice in Cats (애엽(艾葉) (Artemisia asiatica Nakai)의 혈압강하작용(血壓降下作用))

  • Kim, Yun-Ho;Shin, Hong-Kee;Kim, Kee-Soon
    • The Korean Journal of Physiology
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    • v.15 no.2
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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.

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Chromosome Aberration Test of Water Extract of Decaffeined Green Tea using Supercritical Carbon Dioxide with Mammalian cell line (초임계이산화탄소를 이용한 탈카페인녹차 열수추출물의 포유동물 세포주를 이용한 염색체이상시험)

  • Koo, Yun-Chang;Lee, Hyun-Sun;Park, Byung-Gyu;Kim, Eun-Jin;Lee, Sun-Joo;Kim, Kyoung-Hoen;Kim, Young-Suk;Chung, Young-Shin;Lee, Kwang-Won
    • Environmental Mutagens and Carcinogens
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    • v.26 no.4
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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.

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