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Quantitative Determination of Bioactive Compounds in Some Artemisia capillaris by High-Performance Liquid Chromatography

  • Kim, Sang-Won;Kim, Hyeong-Woo;Woo, Mi-Hee;Lee, Jae-Hyun;Choi, Jae-Sue;Min, Byung-Sun
    • Natural Product Sciences
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    • v.16 no.4
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    • pp.233-238
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    • 2010
  • In order to facilitate the quality control of Artemisia capillaris, a simple, accurate and reliable HPLC method was developed for the simultaneous determination of the six bioactive compounds: scopolin (1), chlorogenic acid (2), 2,4-dihydroxy-6-methoxyacetophenone 4-glycoside (3), hyperoside (4), isorhamnetin 3-Orobinobioside (5), and scoparone (6), which were selected as the chemical markers of A. capillaris. Separation was achieved on an Agilent Eclipse XDB-C18 column with a gradient solvent system of 0.1% trifluoroacetic acid aqueous-acetonitrile at a flow-rate of 1.0 mL/min and detected at 254 nm. All six calibration curves showed good linearity ($R^2$ > 0.998). A simple reversed phase HPLC method was developed for extracting pharmacologically active compounds scopolin, chlorogenic acid, 2,4-dihydroxy-6-methoxyacetophenone 4-glycoside, hyperoside, isorhamnetin 3-O-robinobioside, and scoparone from A. capillaris using a binary gradient of acetonitrile : 0.1% trifluoroacetic acid with UV detection at 254 nm. The scopolin (1), chlorogenic acid (2), 2,4-dihydroxy-6-methoxyacetophenone 4-glycoside (3), hyperoside (4), isorhamnetin 3-O-robinobioside (5), and scoparone (6) contents of the herb of A. capillaris collected from fifteen district markets in Korea were 0.00~0.90 mg/g, 0.06~7.29 mg/g, 0.06~0.91 mg/g, 0.07~5.05 mg/g, 0.42~13.11 mg/g, and 1.11~29.82 mg/g, respectively. The results demonstrated that this method is simple and reliable for the quality control of A. capillaris.

Quantitative Determination and Antioxidant Effects of Cheonwangbosimdan (LC-MS/MS를 이용한 천왕보심단(天王補心丹)의 함량분석 및 항산화 효능 연구)

  • Seo, Chang-Seob;Kim, Ohn Soon;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
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    • v.45 no.4
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    • pp.300-314
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    • 2014
  • An ultra-performance liquid chromatography-electrospray ionization-mass spectrometer (UPLC-ESI-MS) method was established for the simultaneous quantification of eighteen marker compounds in traditional Korean formula, Cheonwangbosimdan (CWBSD). In addition, we evaluated the antioxidant effects of CWBSD. Eighteen marker components were separated on a UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) and kept at $45^{\circ}C$ by gradient elution with 0.1% (v/v) formic acid in water and acetonitrile as mobile phase. The flow rate was 0.3 mL/min and the injection volume was $2.0{\mu}L$. The antioxidant activities of CWBSD were assessed by measuring free radical scavenging activities on 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The calibration curves of all analytes showed good linearity (correlation coefficient ${\geq}0.9937$) within the test ranges. The limits of detection and quantification for the 18 marker compounds were 0.01-4.71 ng/mL and 0.03-14.13 ng/mL, respectively. The contents of the 18 compounds in CWBSD extract ranged from none to $1701.00{\mu}g/g$. The CWBSD showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction ($RC_{50}$) against ABTS and DPPH radicals were $149.42{\mu}g/mL$ and $339.24{\mu}g/mL$.

Quantitative Analysis of the Fifteen Constituents in Hyangso-San by LC-MS/MS (LC-MS/MS를 이용한 향소산 중 15종 성분의 정량분석)

  • Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
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    • v.47 no.4
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    • pp.381-388
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    • 2016
  • Hyangso-san is a traditional herbal medicine that consists of the seven herbal medicines, Cyperi Rhizoma, Perillae Folium, Atractylodis Rhizoma, Citri Unshius Pericarpium, Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma Crudus, and Allii Fistulosi Bulbus. Hyangso-san has long been clinically used to treat the influenza, including headache, ferver, chills, and pantalgia. In this study, we were performed the simultaneous analysis of the 15 marker compounds (liquiritin apioside, liquiritin, ferulic acid, naringin, hesperidin, rosmarinic acid, liquiritigenin, kaempferol, glycyrrhizin, nobiletin, 6-gingerol, elemicin, atractylenolide III, nootkatone, and atractylenolide I) in Hyangso-san using liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS/MS). Column for the separation of the 15 ingredients was used a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) at $45^{\circ}C$ by using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient condition. Identifications of all analytes were performed using a Waters ACQUITY TQD LC-MS/MS system. The flow rate and injection volume were 0.3 mL/min and $2.0{\mu}L$, respectively. Correlation coefficient of the calibration curve was ${\geq}0.9958$. The values of limits of detection and quantification of the 15 components were 0.002-4.29 and 0.01-12.88 ng/mL, respectively. The result of an analysis using the established LC-MS/MS method, kaempferol and atractylenolide I were not detected, while other 13 compounds were 0.08-56.87 mg/g in lyophilized Hyangso-san sample.

Quantification of Diesel in Soils using the Partitioning Tracer Method with Two-dimensional Soil Box (분배성 추적자 기법을 이용한 디젤 오염 토양의 정량적 오염도 평가에 관한 2차원 토조 실험 연구)

  • Rhee, Sung-Su;Lee, Gwang-Hun;Park, Jun-Boum
    • Journal of Soil and Groundwater Environment
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    • v.15 no.1
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    • pp.66-72
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    • 2010
  • The partitioning tracer method is to estimate the residual saturation of nonaqueous phase liquid (NAPL) in soils by analyzing tracer's retardation induced by the reversible partitioning of tracer with NAPL. This study is to estimate the residual diesel saturation in soils using the partitioning tracer method. Two-dimensional soil box was used to represent the 2-dimensional flows of groundwater and tracer solution in the saturated aquifer, and the soil box was filled with soil and then saturated with water. The residual diesel saturation was induced in saturated soil, and the partitioning tracer method was applied. The results from batch-partitioning experiment indicated that the diesel-water partitioning was linear with respect to tracer's concentration, and the partition coefficient of tracer between diesel and water was measured by their linearities. The groundwater flow in the saturated aquifer was simulated in the 2-dimensional soil box, and the residual diesel contamination was visually identified. The results from the partitioning tracer method with or without diesel in soils confirmed that 4-methyl-2-pentanol, 2-ethyl-1-butanol and 1-hexanol, can be used as a detecting method for diesel contamination. By the accuracies of estimations for diesel contamination using the partitioning tracer method, 2-ethyl-1- butanol showed the highest accuracy with 83%.

Decomposition of Aqueous Anatoxin-a Using Underwater Dielectric Barrier Discharge Plasma Created in a Porous Ceramic Tube (다공성 세라믹관내에서 생성되는 수중 유전체 장벽 방전 플라즈마를 이용한 아나톡신-a의 분해)

  • JO, Jin-Oh;Jwa, Eunjin;Mok, Young-Sun
    • Journal of Korean Society of Water and Wastewater
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    • v.30 no.2
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    • pp.167-177
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    • 2016
  • This work investigated the decomposition of aqueous anatoxin-a originated from cyanobacteria using an underwater dielectric barrier discharge plasma system based on a porous ceramic tube and an alternating current (AC) high voltage. Plasmatic gas generated inside the porous ceramic tube was uniformly dispersed in the form of numerous bubbles into the aqueous solution through the micro-pores of the ceramic tube, which allowed an effective contact between the plasmatic gas and the aqueous anatoxin-a solution. Effect of applied voltage, treatment time and the coexistence of nutrients such as $NO_3{^-}$, $H_2PO_4{^-}$ and glucose on the decomposition of anatoxin-a was examined. Chemical analyses of the plasma-treated anatoxin-a solution using liquid chromatography-mass spectrometry (LC-MS) and ion chromatography (IC) were performed to elucidate the mineralization mechanisms. Increasing the voltage improved the anatoxin-a decomposition efficiency due to the increased discharge power, but the energy required to remove a given amount of anatoxin-a was similar, regardless of the voltage. At an applied voltage of 17.2 kV (oxygen flow rate: $1.0L\;min^{-1}$), anatoxin-a at an initial concentration of $1mg\;L^{-1}$ (volume: 0.5 L) was successfully treated within 3 min. The chemical analyses using LC-MS and IC suggested that the intermediates with molecular weights of 123~161 produced by the attack of plasma-induced reactive species on anatoxin-a molecule were further oxidized to stable compounds such as acetic acid, formic acid and oxalic acid.

PAHs Formation Characteristics and Fullerenes $(C_{60},\;C_{70})$ Synthesis in a Low-Pressure $C_6H_6/Ar/O_2$ Flame (저압 $C_6H_6/Ar/O_2$ 화염에서 PAHs 생성 특성 및 플러렌$(C_{60},\;C_{70})$ 합성에 대한 연구)

  • Lee, G.W.;Kim, Y.W.;Hwang, J.;Jrung, J.;Choi, M.
    • Journal of the Korean Society of Combustion
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    • v.7 no.4
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    • pp.36-44
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    • 2002
  • Carbon molecules with closed-cage structures are called fullerenes $(C_{60},\;C_{70})$, whose applications include super-conductors, sensors, catalysts, optical and electronic device, polymer composites, and biological and medical materials. The synthesis of fullerenes has been recently studied with low-pressure benzene/argon/oxygen flames. The formation of fullerene is known as molecular weight growth processes of PAHs (polycyclic aromatic hydrocarbon). This study presents results of PAHs and fullerene measurements performed in a low-pressure benzene/argon/oxygen normal co-flow laminar diffusion flame. Through the central tube of the burner, benzene vapors carried by argon are injected. The benzene vapors are made in a temperature-controlled bubbler. The burner is located in a chamber, equipped with a sampling system for direct collection of condensable species from the flame, and exhausted to a vacuum pump. Samples of the condensable are analyzed by HPLC (High Performance Liquid Chromatography) to determine the yields of PAHs and fullerene. Also, we computed mole fraction of fullerene and PAHs in a nearly sooting low pressure premixed, one-dimensional benzene/argon/oxygen flame (equivalence ratio ${\Phi}=2.4$, pressure=5.33kPa). The object of computation was to investigate the formation mechanism of fullerenes and PAHs. The computations were performed with CHEMKIN/PREMIX. As a result of this study, fullerenes were synthesized in a low pressure (20torr) $C_6H_6/Ar/O_2$ flames and the highest concentration of fullerene was detected just above the visible surface of a flame.

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Effect of Nanotube Length on Rheological Characteristics of Polystyrene/Multi-walled Carbon Nanotube Nanocomposites Prepared by Latex Technology (라텍스 기법으로 제조한 폴리스티렌/다중벽 탄소나노튜브 나노복합재료의 나노튜브 길이가 유변학적 특성에 미치는 영향)

  • Woo, Dong-Kyun;Noh, Won-Jin;Lee, Seong-Jae
    • Polymer(Korea)
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    • v.34 no.6
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    • pp.534-539
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    • 2010
  • Polystyrene (PS)/multi-walled carbon nanotube (MWCNT) nanocomposites were prepared via latex technology and the effect of nanotube length on rheological properties were investigated. Monodisperse PS particle was synthesized by the emulsifier-free emulsion polymerization and two types of MWCNTs were used after surface modification to improve dispersion state and to remove impurities. Final nanocomposites were prepared by the freeze-drying process after dispersing the PS particles and the surface-modified MWCNTs in a ultrasonic bath. The effects of MWCNT content and nanotube length on rheological properties were evaluated by imposing the small-amplitude oscillatory shear flow. The PS/MWCNT nanocomposites showed that rheological properties were enhanced as the amount and length of MWCNT increased. It is speculated that the rheological characteristics of nanocomposites change from liquid-like to solid-like as the MWCNT amount increases, and the critical concentration to achieve network structure decreases as the nanotube length increases.

Analytical Optimum of Ginsenosides according to the Gradient Elution of Mobile Phase in High Performance Liquid Chromatography (HPLC의 이동상 용매조건에 따른 인삼 Ginsenoside 분석)

  • Park, Ji-Yeong;Won, Jun-Yeon;Lee, Chung-Yeol
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.3
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    • pp.215-219
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    • 2007
  • This study was conducted to analyze not only for the quality guaranteed of red ginseng but also for the minor ginsenosides. Although several studies have reported to analyze ginseng saponins, those were focused to major saponins, including 6 to 7 ginsenosides. As increase of interest in medicinal effect of ginseng products, anasis of various ginsenosides in both red and white ginseng are strongly demanded. To perform optital condition of 12 ginsenoside analysis, We controlled HPLC conditions, such as the gradient elution of the mobile phase. We found the adequate separation method for 12 ginse-nosides. The optimum condition was as following : H$_2$O/CH$_3$CN ratios were 82/18, 70/30, 55/45 and 50/50, respectively. Sol-vent flow rate was 1.00 ma/min. Column temperature was kept to 35$^{\circ}$C. UV detector was set to 203 nm.

Effects of Bojeongjeongcheon-tang on Cytokines and Immunoglobulin E in B Cells (보정정천탕의 Cytokine 및 IgE에 대한 조절효과)

  • 권혁성;정주호;김성훈;정승기
    • The Journal of Korean Medicine
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    • v.25 no.2
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    • pp.51-66
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    • 2004
  • Objectives : To evaluate experimentally the clinical effect of Bojeongjeongcheon-tang, we observed the cytokines ($IL-1{\beta}$/TEX>, IL-4, IL-5, IL-6, IL-10, TNF-{\alpha},{\;}TGF-{\beta},{\;}IFN-{\gamma}$) and what effect they have on IgE in B cells of a rat. Methods : First of all, we extracted the spleens of healthy Balb/c mice and separated B cells from them. These B cells were cultured with anti-CD40 mAb (500 ng/ml), rmIL-4 (500 U/ml), Bojeongjeongcheon-tang (100 ug/ml, 10 ug/ml, 1 ug/ml). We used rmiL-10 (50 ng/ml) as a control group. Furthermore, we analyzed the expression of IgE, CD23, CD69 and the coherence of HRF in B cells using a flow cytometer. We also analyzed the cytokine gene expression in B cells by reverse transcriptase-PCR. We also measured B cells proliferation using the Liquid Scintillation Counter. Results : In this study, the Bojeongjeongcheon-tang treated group showed a tendency to decrease depending on the density compared with the control group in the expression of IgE+, CD23+, CD69, HRF. All of the Bojeongjeongcheon-tang treated group showed inhibitory effects with $IL-1{\beta}$, IL-4, IL-5 and proliferating effects with IL-6, IL-10, and $IFN-{\gamma}$ on cytokines transcript expression depending on the density. Meanwhile, $TNF-{\alpha}$ increased in all density. In IgE production, there was inhibitory effect on Bojeongjeongcheon-tang (both 100 ug/ml and 10 ug/ml) of significance (p < 0.01, p < 0.05). Also in B cell proliferation, the result revealed an inhibitory effect of Bojeongjeongcheon-tang (both 100 ug/ml and 10 ug/ml), of significance (p < 0.001, p < 0.01). Conclusions : This study shows that Bojeongjeongcheon-tang has an inhibitory effect on the production and activity of B cells. Also it inhibited CD23, IL-4 activity and IgE production and activation. It is obvious that Bojeongjeongcheon-tang treats asthma by inhibiting the production of histamine and HRF, IL-5 and proliferating IL-10. Also Bojeongjeongcheon-tang has some preventive effects on bronchial change by inhibiting $TGF-{\beta}$, which stimulates the bronchial transformation.

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Performance Characteristics with Various Fuel Composition and Temperature for an External Type Fuel Pump in LPLi System (LPLi 시스템에서 외장형 펌프의 연료조성 및 온도에 따른 성능특성 연구)

  • Nam, Deok-Woo;Yoon, Jun-Kyu;Lim, Jong-Han
    • Journal of Advanced Marine Engineering and Technology
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    • v.35 no.5
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    • pp.566-575
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    • 2011
  • Since LPG (Liquefied petroleum gas) fuel supply system has an advantage of low emission characteristics, many studies have been conducted. In spite of the advantage of LPG supply system, a higher vapor pressure and lower viscosity than diesel or gasoline fuel may cause unstable running of fuel pump by the deterioration in lubrication performance and chemical reaction with rubber parts than that of diesel and gasoline fuel. Therefore its physical properties can cause the deterioration of durability. In this research, we developed an external type LPG pump which has the advantage of the price competitiveness and the convenient maintenance for LPLi system. The experiments were carried out in order to assess characteristics of the external type fuel pump at different fuel composition and temperature. As a result, there aren't any differences between internal and external type pump performance. It is observed that the same level of efficiency was maintained for both pumps as flow rate was increased with higher fuel temperature and more contents of propane in the fuel. And the pressure difference in LPLi system is maintained at constant with the various fuel compositions and temperatures due to their own characteristics of fuel supply system.