• Title/Summary/Keyword: Ion-pair

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Quantitative determination of inosine 5'-monophosphate dehydrogenase activity in human peripheral blood mononuclear cells by ion-pair reversed-phase high-performance liquid chromatography (이온쌍 역상 HPLC를 이용한 인체 말초혈액단핵구에서 이노신 5'-일인산 탈수소효소 활성의 정량적 측정)

  • Shin, Hye-Jin;Kwon, Soon-Ho;Park, Ji-Myeong;Kwon, Soon-Hyo;Lee, Kyoung-Ryul;Kim, Young-Jin;Lee, Sang-Hoo
    • Analytical Science and Technology
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    • v.23 no.6
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    • pp.531-536
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    • 2010
  • A quantitative analytical method has been established for the measurement of inosine 5'-monophosphate dehydrogenase (IMPDH) activity in human peripheral blood mononuclear cells (PBMCs) by ion-pair reversed-phase high performance liquid chromatography equipped with ultraviolet detection (HPLC/UV). IMPDH is a ${\beta}$-nicotinamide adenine dinucleotide hydrate (NAD+)-dependent dehydrogenase in which the enzyme converts inosine 5'-monophosphate (IMP) into xanthosine 5'-monophosphate (XMP). Its activity was measured by quantifying a HPLC chromatogram corresponding to XMP produced during the incubation of lysed PBMCs with IMP as a substrate and $NAD^+$ as a coenzyme. XMP produced was detected at a wavelength of 260 nm. The mobile phase was composed of a mixture of 37 mM potassium dihydrogen phosphate containing 7 mM tetra-n-butylammonium hydrogen sulfate adjusted to pH 5.5 and methanol (85:15, v/v) with a flow rate of 1 mL/min. The calibration curve was linear ($r^2$=0.999999) in the range of $0.2-50.0\;{\mu}M$ and the limit of quantification (LOQ) was $0.2\;{\mu}M$. The intra- and inter-day precisions were between 0.88-1.47% and 0.85-5.24%, respectively. The intra- and inter-day accuracies were between 98.74-99.99% and 99.95-101.65%, respectively. IMPDH activity in 11 Korean healthy volunteers ranged from 18.29 to 36.60 nmol/h/mg protein (mean = $27.70{\pm}6.28\;nmol/h/mg$ protein).

Optimization of HPLC Method and Clean-up Process for Simultaneous and Systematic Analysis of Synthetic Color Additives in Foods (식품 중 타르색소의 동시분석 및 계통분석을 위한 HPLC 분석조건 및 정제과정 확립)

  • Park, Sung-Kwan;Hong, Yeun;Jung, Yong-Hyun;Lee, Chang-Hee;Yoon, Hae-Jung;Kim, So-Hee;Lee, Jong-Ok
    • Korean Journal of Food Science and Technology
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    • v.33 no.1
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    • pp.33-39
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    • 2001
  • To develop a method for separation process using Sep-pak $C_18$, simultaneous and systematic analysis of 8 permitted and 11 non-permitted synthetic food colors in Korea, optimization of analysis conditions for reverse phase ion-pair high performance liquid chromatography was carried out. For the best result of Sep-pak $C_18$ separation the pH of color standard mixture solution was $5{\sim}6$ and 0.1% HCl-methanol solution were set as eluent. The colors eluated from Sep-pak $C_18$ cartridge were determined and confirmed by high performance liquid chromatography with a photodiode array detector at 420 nm for yellow colors type, at 520 nm for red colors type, at 600 nm for blue and green colors type and at 254 nm for mixed colors. Conditions for HPLC analysis were as follows: column, Symmetry $C_18$ (5 m, 3.9 mm $i.d.{\times}150\;mm$); mobile phase, 0.025 M ammonium acetate (containing 0.01 M tetrabutylammonium bromide) : acetonitrile : methanol (65 : 25 : 10) and 0.025 M ammonium acetate(containing 0.01 M tetrabutylammonium bromide) : acetonitrile : methanol (40 : 50 : 10); flow rate, 1 mL/min. It takes 35 minutes for simultaneaus analysis and 18 minutes for systematic analysis. The detection limits range of each colors were $0.01{\sim}0.05\;{\mu}g/g$.

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Monte Carlo Simulation of the Carbon Beam Nozzle for the Biomedical Research Facility in RAON (한국형 중이온 가속기 RAON의 의생물 연구시설 탄소 빔 노즐에 대한 Monte Carlo 시뮬레이션)

  • Bae, Jae-Beom;Cho, Byung-Cheol;Kwak, Jung-Won;Park, Woo-Yoon;Lim, Young-Kyung;Chung, Hyun-Tai
    • Progress in Medical Physics
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    • v.26 no.1
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    • pp.12-17
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    • 2015
  • The purpose of the Monte Carlo simulation study was to provide the optimized nozzle design to satisfy the beam conditions for biomedical researches in the Korean heavy-ion accelerator, RAON. The nozzle design was required to produce $C^{12}$ beam satisfying the three conditions; the maximum field size, the dose uniformity and the beam contamination. We employed the GEANT4 toolkit in Monte Carlo simulation to optimize the nozzle design. The beams for biomedical researches were required that the maximum field size should be more than $15{\times}15cm^2$, the dose uniformity was to be less than 3% and the level of beam contamination due to the scattered radiation from collimation systems was less than 5% of total dose. For the field size, we optimized the tilting angle of the circularly rotating beam controlled by a pair of dipole magnets at the most upstream of the user beam line unit and the thickness of the scatter plate located downstream of the dipole magnets. The values of beam scanning angle and the thickness of the scatter plate could be successfully optimized to be $0.5^{\circ}$ and 0.05 cm via this Monte Carlo simulation analysis. For the dose uniformity and the beam contamination, we introduced the new beam configuration technique by the combination of scanning and static beams. With the combination of a central static beam and a circularly rotating beam with the tilting angle of $0.5^{\circ}$ to beam axis, the dose uniformity could be established to be 1.1% in $15{\times}15cm^2$ sized maximum field. For the beam contamination, it was determined by the ratio of the absorbed doses delivered by $C^{12}$ ion and other particles. The level of the beam contamination could be achieved to be less than 2.5% of total dose in the region from 5 cm to 17 cm water equivalent depth in the combined beam configuration. Based on the results, we could establish the optimized nozzle design satisfying the beam conditions which were required for biomedical researches.

Determination of Pantothenic acid in Fortified Foods by HPLC (시판 영양강화식품중 판토텐산의 분석)

  • 최윤주;장재희;박혜경;박건상;구용의;황인경;김대병
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.2
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    • pp.381-385
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    • 2004
  • This study was conducted to develop an HPLC method for determining pantothenic acid in fortified foods which has typically been determined by microbiological assay (MBA) according to AOAC and Korean Food Code approved methods. Pantothenic acid was determined by reversed-phase ion-pair HPLC using UV absorption (200 nm) after extraction with 20 mM potassium phosphate solution by sonication. The recovery of spiked samples and detection limit (LOD) by HPLC were 83.5∼109.6% and 0.5 ppm (mg/kg), respectively. The LOD of the microbiological assay (MBA) was much lower than that of HPLC. The concentrations of pantothenic acid analyzed in all tested samples (n=13) confirmed compliance with declared label claims. The range of recovery ratio by the HPLC method when compared to the microbiological assay was 91.9∼117.6%. There was not significant difference (p<0.01) between the HPLC and MBA methods and the equation of the regression curve was y=1.1428x-0.2269 (r=0.9842). This proposed HPLC method for determining pantothenic acid appears to be suitable for determining pantothenic acid concentrations above 0.25 mg/100 g in fortified foods.

Increased Sensitivity of Carbon Nanotube Sensors by Forming Rigid CNT/metal Electrode

  • Park, Dae-Hyeon;Jeon, Dong-Ryeol
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.08a
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    • pp.348-348
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    • 2011
  • Carbon nanotube (CNT) field effect transistors and sensors use CNT as a current channel, of which the resistance varies with the gate voltage or upon molecule adsorption. Since the performance of CNT devices depends very much on the CNT/metal contact resistance, the CNT/electrode contact must be stable and the contact resistance must be small. Depending on the geometry of CNT/electrode contact, it can be categorized into the end-contact, embedded-contact (top-contact), and side-contact (bottom-contact). Because of difficulties in the sample preparation, the end-contact CNT device is seldom practiced. The embedded-contact in which CNT is embedded inside the electrode is desirable due to its rigidness and the low contact resistance. Fabrication of this structure is complicated, however, because each CNT has to be located under a high-resolution microscope and then the electrode is patterned by electron beam lithography. The side-contact is done by depositing CNT electrophoretically or by precipitating on the patterned electrode. Although this contact is fragile and the contact resistance is relatively high, the side-contact by far has been widely practiced because of its simple fabrication process. Here we introduce a simple method to embed CNT inside the electrode while taking advantage of the bottom-contact process. The idea is to utilize a eutectic material as an electrode, which melts at low temperature so that CNT is not damaged while annealing to melt the electrode to embed CNT. The lowering of CNT/Au contact resistance upon annealing at mild temperature has been reported, but the electrode in these studies did not melt and CNT laid on the surface of electrode even after annealing. In our experiment, we used a eutectic Au/Al film that melts at 250$^{\circ}C$. After depositing CNT on the electrode made of an Au/Al thin film, we annealed the sample at 250$^{\circ}C$ in air to induce eutectic melting. As a result, Au-Al alloy grains formed, under which the CNT was embedded to produce a rigid and low resistance contact. The embedded CNT contact was as strong as to tolerate the ultrasonic agitation for 90 s and the current-voltage measurement indicated that the contact resistance was lowered by a factor of 4. By performing standard fabrication process on this CNT-deposited substrate to add another pair of electrodes bridged by CNT in perpendicular direction, we could fabricate a CNT cross junction. Finally, we could conclude that the eutectic alloy electrode is valid for CNT sensors by examine the detection of Au ion which is spontaneously reduced to CNT surface. The device sustatined strong washing process and maintained its detection ability.

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Pressure Effects on the Ionic Dissociation of $[Co(en)]_3Cl_3\;and\;[Co(NH_3)_6Cl_3$ in Aqueous Solutions (수용액에서 $[Co(en)_3]Cl_3$$[Co(NH_3)_6]Cl_3$착물의 이온 해리에 미치는 압력의 영향)

  • Jong Jae Chung;Rho Byung Gill
    • Journal of the Korean Chemical Society
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    • v.30 no.4
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    • pp.335-450
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    • 1986
  • The thermodynamic dissociation constants of $[Co(en)]_3Cl_3\;and\;[Co(NH_3)_6Cl_3$ in aqueous solutions at $25{\circ}$ and at the pressure from 1 bar to 2000 bars were measured by conductometric method. The dissociation constants were increased with pressure elevation because of volume decreasing by the formation of charged ions during dissociation reaction. That is, the $pK^T$ values of $[Co(en)]_3Cl_3$ were 2.16 for I bar, 2.08 for 500 bars, 2,08 for 1000 bars, 2.05 for 1500 bars and 2.03 for 2000 bars, respectively and those of $[Co(NH_3)_6Cl_3$ were 2.02 for 1 bar 1.96 for 500 bars, 1.90 for 1000 bars, 1. 88 for 1500 bars and 1. 87 for 2000 bars, respectively. Comparing and analyzing the values of Stokes' radii and $K^T$, the formation of ion pair compound was affected by not only the electrostatic interaction, but also the Interal Conjugate Base(ICB), which was increased by the elevation of the pressure.

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