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HPLC Analysis of Marker Substances in Taeumin Gamijowi-tang (태음인(太陰人) 가미조위탕(加味調胃湯)의 HPLC 분석)

  • Hwang, Young-Myung;Yu, Young-Beob;Kim, Jong-Youl;Koh, Byung-Hee;Lee, Eui-Ju
    • Journal of Sasang Constitutional Medicine
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    • v.18 no.1
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    • pp.147-154
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    • 2006
  • 1. Objectives This study was aimed to evaluate marker substances in Gamijowi-tang by high performance liquid chromatography(HPLC) 2. Methods The seperation was performed an AquasilC18(4.6X250mm)column by gradient elution with 0.1N H3PO4 in H2O - acetonitrile(0min 100:10,20min60:40, 40min 80:20, 60min 100:0) as the mobile phase at a flow-rate of 1.0 ml/min with detection at 190-800nm 3. Results Ephedrine and schizandrin were established as marker substances in Gamijowi-tang and its qualitative analysis was carried out. Analyzed was similarity of marker substances between in standard article and in Gamijowi-tang. The result of the analysis was that ephedrine and schizadrin showed 0.9999 of similarity, which meant they were really ephedrine and schizadrin. In addition, ephedrin content was 12.16mg, pseudoephedrine conrenr was 5.09mg, and schizandrin content was 26.6mg in 76g of Gamijowi-tang. 4. Conclusions Qualitative and quantitative analyses of ephedrine and schizandrin in Gamijowi-tang were conducted, and it was identified thar they could be used as marker substances in standardizing quality of Gamijowi-rang.

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Study on the Pro-apoptotic Effect of Artemisiae Capillaris Herba Extracted with Ethanol on Biliary Tract Cancer Cell Line, SNU-1196 (인진(茵蔯)의 에탄올 추출물이 담도암 세포주 SNU-1196의 apoptosis에 미치는 효과에 관한 연구)

  • Lee, Kyung-Wook;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.33 no.4
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    • pp.587-598
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    • 2012
  • Objectives : This study was performed to elucidate the pro-apoptotic effect of Artemisiae capillaris herba extracted with ethanol on biliary tract cancer cells. Materials and Methods : The biliary tract cancer cell line SNU-1196 was used in this study. Cells were treated with different concentrations of Artemisiae capillaris herba for 24, 48 and 72 hours. After the treatment, cell viability, apoptosis, caspase activities and the mRNA expressions of the Bcl-2, Bax, P53, and P21 were measured by using MTT assay, cell cycle analysis, apoptosis assay, and RT-PCR. The cell cycle analysis was done by flow cytometry and apoptosis assay by cell death detection ELISA kit. Results : Artemisiae capillaris herba inhibited proliferation of SNU-1196 in long-time culture group with dose-dependent manner. All cells treated with Artemisiae capillaris herba showed increased apoptosis with dose- and time-dependent manner. Exposure of SNU-1196 to Artemisiae capillaris herba induced caspase-3 activation. However, apeoptosis was blocked when SNU-1196 was treated together with the pan-caspase inhibitor Z-VAD-FMK and the caspase-3 inhibitor Z-DEVE-FMK. After the treatment of Artemisiae capillaris herba, the mRNA expressions of caspase -3, -8, -9, p53, and p21 was increased in all cells. Artemisiae capillaris herba resulted in a significant decrease in Bcl-2 and an increase in Bax mRNA levels. Conclusions : These results suggest that Artemisiae capillaris herba would be beneficial in the treatment of biliary tract cancer.

Preparation of L-cysteine Salicylaldehyde Schiff-base Modified Macroporous Polystyrene Resin and Its Application to Determination of Trace Cadmium and Lead in Environmental Water Samples

  • Xie, Fazhi;Zhang, Fengjun;Xuan, Han;Ge, Yejun;Wang, Yin;Li, Guolian;Zhu, Lei;Oh, Won-Chun
    • Journal of the Korean Ceramic Society
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    • v.51 no.5
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    • pp.472-476
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    • 2014
  • In this work, a new method that utilizes L-cysteine salicylaldehyde Schiff-base modified macroporous polystyrene resin (PS-CSC) as an effective sorbent has been developed for preconcentration of trace cadmium and lead in environmental water samples. The effect of pH, the contact time, the elution conditions, the flow rate, the initial concentration of target metal ions, and the effects of interfering ions on the preconcentration of the analytes were investigated. The maximum adsorption capacity of PS-CSC under optimum conditions for cadmium and lead were found to be 6.03 - 18.17 mg/g and 12.58 - 36.13 mg/g when the initial concentration of metal ions between 5.0 - 90 mg/L. The limits of detection for cadmium and lead were 2.46 ng/L and $0.52{\mu}g/L$, with a preconcentration factor of 200. The developed method has been validated by analyzing certified reference material and successfully applied for the enrichment and determination of trace cadmium and lead from environmental water samples.

In vitro Anti-allergic Effects of Bangpungtongseong-san in Human Keratinocytes and Primary Mouse Splenocytes (간 각질 세포 및 마우스 비장 세포를 이용한 방풍통성산의 항알러지 효능 연구)

  • Jeong, Soo-Jin;Lee, Mee-Young;Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.29 no.2
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    • pp.168-173
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    • 2015
  • Bangpungtongseong-san(BPTSS, Fangfengtongsheng-san in Chineses) is a traditional herbal formula comprising 18 medicinal herbs. In the present study, we performed the simultaneous analysis for four compounds of BPTSS and examined anti-allergic effects in human keratinocytes and mouse splenocytes. The column for separation of four compounds was used Luna C18 column and maintained at 40℃. The mobile phase for gradient elution consisted of two solvent systems. The analysis was carried out at a flow rate of 1.0 mL/min with PDA detection at 254 and 280 nm. To evaluate production and expression of Th2 chemokines, ELISA and RT-PCR were conducted in tumor necrosis factor (TNF)-α and interferon (IFN)-γ-stimulated HaCaT cells with or without BPTSS or silymarin, a positive control for skin inflammation. To measure Th2 cytokines, primary mouse splenocytes were treated with BPTSS and performed ELISA for interleukin (IL)-4, 5, 13. Calibration curves were acquired with r2>0.9999. The contents of geniposide, liquiritin, baicalin, and glycyrrhizin in BPTSS were 5.06 ㎎/g, 7.33 ㎎/g, 27.56 ㎎/g, and 7.81 ㎎/g, respectively. BPTSS reduced TARC and RANTES production and mRNA expression in TNF-α and IFN-γ-treated HaCaT cells. BPTSS inhibited IL-4, 5, and 13 production in mouse splenocytes. Our data will be a helpful information to upgrade quality control and anti-allergic effects of BPTSS.

Simultaneous Determination of Gumiganghwal-tang and Its Anti-atherosclerotic Effect (구미강활탕의 성분 분석 및 LDL 산화억제 효능 연구)

  • Kim, Ohn Soon;Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • Journal of Society of Preventive Korean Medicine
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    • v.17 no.3
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    • pp.165-176
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    • 2013
  • Objective : The purpose of this study was to establish the simultaneous analysis for six compounds in Gumiganghwal-tang (GMGHT, Jiuweiqianghuo-tang) and to investigate the anti-atherosclerotic effects of GMGHT in vitro. Methods : The column for separation of six compounds was used Luna $C_{18}$ column and maintained at $40^{\circ}C$. The mobile phase for gradient elution consisted of two solvent systems, 1.0% acetic acid in water and 1.0% acetic acid in acetonitrile. The analysis was carried out at a flow rate of 1.0 mL/min with pothodiode array (PDA) detection at 254, 280, and 320 nm. The injection volume was 10 ${\mu}L$. The antioxidant activities of GMGHT were evaluated by measuring free radical scavenging activities on 2,2'-Azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The inhibitory effects on low-density lipoprotein (LDL) oxidation were evaluated by the formation of thiobarbituric acid relative substances (TBARS), relative electrophoretic mobility (REM), and fragmentation of apolipoprotein B (ApoB)-100. Results : Calibration curves were acquired with $r^2{\geq}0.9998$. The contents of liquiritin, ferulic acid, baicalin, baicalein, glycyrrhizin, and wogonin in GMGHT were 1.784, 1.693, 37.899, 0.258, 1.869, and 0.034 mg/g, respectively. The GMGHT showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction ($RC_{50}$) against ABTS and DPPH radicals were 72.51 ${\mu}g/mL$ and 128.49 ${\mu}g/mL$. Furthermore, GMGHT reduced the oxidation properties of LDL induced by $CuSO_4$. Conclusion : HPLC-PDA is considered as an available and convenient method for quality control and standardization of GMGH and GMGHT has potentials on anti-atherosclerosis by anti-oxidative effect and suppressive effect on LDL oxidation.

Application of Polyurethane Foam Loaded with a Schiff Base Ligand for Determination of Trace Amounts of Copper in Water Samples by Flame Atomic Absorption Spectroscopy

  • Moftakhar, Mahdieh Koorehpazan;Yaftian, Mohammad Reza;Nahaei, Samaneh;Zamani, Abbas Ali
    • Journal of the Korean Chemical Society
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    • v.58 no.3
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    • pp.283-288
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    • 2014
  • A column solid phase extraction procedure based on modified polyurethane foam (PUF) by a newly synthesized Schiff base ionophore, named 2,2'-{iminobis[propane-3,1-diylnitrilo(1E)prop-1-yl-1-ylidene]}diphenol, was developed for preconcentration step of trace amounts determination of copper ions in water samples by atomic absorption flame spectroscopy. The influence of parameters on the adsorption process such as sample pH, amount of modified PUF packed in the column, type and volume of stripping reagent and its flow rate were investigated and optimized. Under optimum experimental conditions, the calibration graph was linear in a relatively wide range ($0.005-210{\mu}g/ml$) with a limit of detection $0.002{\mu}g/ml$ of copper. The proposed method allows achieving to a concentration factor of >133. The capacity of a column (1.6 cm i.d.) packed by 6 g of PUF modified by 12 mg of the Schiff base was found to be $247.7({\pm}2.1){\mu}g$ of copper. It was found that the adsorption process was highly selective towards copper ions with respect to some associated metal ions. The presented procedure was successfully applied for determination of copper in some water samples.

Liquid Chromatography-Tandem Mass Spectrometric Determination of Geniposide in Rat Plasma and its Pharmacokinetic Application

  • Jin, Ming Ji;Kim, In Sook;Kim, Dong-Hyun;Yoo, Hye Hyun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.9
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    • pp.2760-2764
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    • 2013
  • Geniposide is a biologically active ingredient of gardenia fruit. A liquid chromatography-tandem mass spectrometric method was developed and validated for the determination of geniposide in rat plasma. The plasma samples were pretreated by solid-phase extraction and introduced into a BDS Hypersil $C_{18}$ column ($50{\times}2.1mm$, $5{\mu}m$) for chromatographic separation. The mobile phase consisted of 0.1% formic acid and 0.1% formic acid in acetonitrile, and gradient elution was performed at a flow rate of 0.25 mL/min. For mass spectrometric detection, multiple reaction monitoring was performed via an electrospray ionization source in positive mode. The calibration curve for geniposide was linear ($r^2=0.997$) in the concentration range of $0.005-1{\mu}g/mL$. The intra- and inter-day accuracies and precisions fulfilled the required criteria (${\pm}15%$). The developed method was subsequently used for pharmacokinetic analysis of geniposide after oral administration to rats at a dose of 50 mg/kg. The mean maximum plasma concentration of geniposide was $0.68{\pm}0.29{\mu}g/mL$ at $0.44{\pm}0.13h$, and the mean area under the plasma concentration versus time curve was $1.46{\mu}g{\cdot}h/mL$.

Development of HPLC method for differentiation of three parts of mulberry tree

  • Eom, Ji Hyun;Vu, Thi Phuong Duyen;Cai, Linxi;Zhao, Yan;Li, Hong Xu;Yang, Seo Young;Kim, Young Ho;Kim, Seok Jin;Cho, Hyun So;Bao, Haiying;Chem, Jianbo;Kim, Kyung Tae;Kang, Jong Seong
    • Analytical Science and Technology
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    • v.30 no.3
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    • pp.130-137
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    • 2017
  • The leaves (Mori Folium; MF), branches (Mori Ramulus; MR), and root bark (Mori Cortex Radicis; MCR) of the mulberry tree have been used as therapeutic herbs for centuries. Existing analytical methods were developed specifically for different parts of the tree and cannot be applied to samples containing a mixture of tree parts. Such method specialization is time-consuming and requires separate identification and quality control of each tree part. This report describes an HPLC method for the simultaneous quality control and discrimination of MF, MR, and MCR using four marker compounds: rutin, kuwanon G, oxyresveratrol, and morusin. An Optimapak $C_{18}$ column ($4.6{\times}250mm$, $5{\mu}m$) was used with a gradient elution of 0.1 % formic acid in water and acetonitrile. The flow rate was 1.0 mL/min and the detection wavelength was 270 nm. In quantitative analyses of the three parts, rutin (0.11 % w/w) was detected only in MF. The oxyresveratrol content (0.12 % w/w) was highest in MR. Kuwanon G (0.33 % w/w) and morusin (0.18 % w/w) were higher in MCR than in other parts. The HPLC method given herein can be used to simultaneously classify and quantify three herbal medicines from the mulberry tree.

A Study of Penetration Depth into Ceiling Materials containing Asbestos according to Dilution Rate of Scattering Prevention Agent (석면 함유 천장재의 안정화제 희석에 따른 침투깊이 연구)

  • Shin, Hyungyoo;Choi, Youngkue;Jeon, Boram;Ha, Jooyeon
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.25 no.1
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    • pp.82-88
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    • 2015
  • Objectives: This study is designed to analyze the penetration performance into ceiling materials containing asbestos of scattering prevention agents and investigate the change in penetration depth and viscosity according to the dilution rate of anti-scattering agents diluted with distilled water. Methods: Five different types of scattering prevention agents were spread on plate-type asbestos ceiling materials. The penetration depth of each coated ceiling material was measured by energy dispersive spectroscopy (EDS) analysis, based on X-ray fluorescence (XRF) results of the non-coated ceiling materials. Test equipment installed the ceiling materials and 60 minutes were collected at a flow rate of $10{\ell}/min$ at a filter of 25 mm. Results: An EDS analysis of the cross-section of ceiling materials constructed with a scattering prevention agent revealed that potassium is detected in the process of penetrating hardener solidification and this element could be an indicator for infiltration. When anti-scattering agents with different viscosities were constructed and the penetration depth was analyzed by potassium detection assessment using EDS, the depth results with viscosities of 5.0, 2.5, and 1.9 cP were 98.5, 103, and $147{\mu}m$, respectively. Penetration performance improved with decrease in viscosity. Conclusions: For asbestos ceiling materials, it is concluded that a higher dilution rate of the scattering prevention agent leads to lower viscosity, and hence a deeper penetration depth from $156{\mu}m$ to 3 mm. The asbestos anti-scattering properties according to the penetration depth will be confirmed through further study.

Branch Misprediction Recovery Mechanism That Exploits Control Independence on Program (프로그램 상의 제어 독립성을 이용한 분기 예상 실패 복구 메커니즘)

  • Yoon, Sung-Lyong;Lee, Won-Mo;Cho, Yeong-Il
    • Journal of KIISE:Computer Systems and Theory
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    • v.29 no.7
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    • pp.401-410
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    • 2002
  • Control independence has been put forward as a new significant source of instruction-level parallelism for superscalar processors. In branch prediction mechanisms, all instructions after a mispredicted branch have to be squashed and then instructions of a correct path have to be re-fetched and re-executed. This paper presents a new branch misprediction recovery mechanism to reduce the number of instructions squashed on a misprediction. Detection of control independent instructions is accomplished with the help of the static method using a profiling and the dynamic method using a control flow of program sequences. We show that the suggested branch misprediction recovery mechanism improves the performance by 2~7% on a 4-issue processor, by 4~15% on an 8-issue processor and by 8~28% on a 16-issue processor.