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Separation Characteristics of Whey Protein by High Performance Membrane Chromatography (고성능 막 크로마토그래피에 의한 유청 단백질의 분리특성)

  • 홍승범;노경호
    • KSBB Journal
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    • v.16 no.6
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    • pp.533-537
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    • 2001
  • ${\alpha}$-lactalbumin and ${\beta}$-lactoglobulin in whey proteins were separated by high performance membrane chromatography (HPMC). The separation mechanism involved anion-exchange, and the stationary phase was anion CIM (Convective Interaction Media) DEAE, QA disk and cation exchanger SO$_3$(16${\times}$3 mm). Two types of mobile phase were used, buffer A (20 mM Tris-HCI, pH 7.3) and buffer B(buffer A + 1 M NaCl), As the amount of NaCl dissolved in buffer linearly increased, which enabled a gradient elution mode. The optimum mobile phase and operating condition (Buffer A/Buffer B = 100/0 - 30/70 vol%, gradient time 1 min, 30/70 - 10/90 vol.%, gradient time 2 min) were experimentally determined. In this experimental condition, ${\alpha}$-lacta1bumin, ${\beta}$-lactoglobulin were separated within 5 min at a mobile phase flow rate of 4 mL/min.

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Quantitative Determination of Bakkenolide D in Petasites japonicus and Farfugium japonicum by HPLC/UV

  • Quilantang, Norman G.;Lee, Ki Ho;Lee, Dong Gu;Lee, Ju Sung;Cho, EunJu;Kim, Hyun Young;Lee, Sanghyun
    • Natural Product Sciences
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    • v.23 no.4
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    • pp.270-273
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    • 2017
  • A quantitative analysis of bakkenolide D in the different parts of Petasites japonicus and Farfugium japonicum was performed by HPLC. A gradient HPLC elution system with a mobile phase consisting of water: acetonitrile solution (20:80 to 0:100 for 45 min) was followed and an INNO $C_{18}$ column was used for the chromatographic separation. The injection volume, flow rate, and UV detection were $10{\mu}L$, 1 mL/min, and 290 nm, respectively. Results show that both species showed the highest amount of bakkenolide D in the roots being 107.203 and 166.103 mg/g for P. japonicas and F. japonicum, respectively. Content analysis on the different parts of both plants displayed remarkably lower values which ranged from 0.403 - 4.419 and 7.252 - 32.614 mg/g for P. japonicas and F. japonicum, respectively. The results show that the roots of both plants are rich in bakkenolide D showing a promising use in the development of nutraceuticals and industrial application of the compound.

Contents of Poncirin and Naringin in Fruit of Poncirus Trifoliata according to Different Harvesting Times and Locations for Two Years (2년 동안 지실의 채취시기 및 지역에 따른 Poncirin과 Naringin의 함량)

  • Lee, Sul-Lim;Seo, Chang-Seob;Kim, Jung-Hoon;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
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    • v.42 no.2
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    • pp.138-143
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    • 2011
  • A high performance liquid chromatography (HPLC) method was developed for simultaneous determination of two major flavonoid glycosides (poncirin and nanringin) in Poncirus trifoliata Raf. by different harvesting times and locations for two years. A SunFire $C_{18}$ column (4.6 mm${\times}$250 mm, 5 ${\mu}M$) was used at $40^{\circ}C$ for the determination of poncirin and naringin. The mobile phase using gradient flow consisted of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. Flow rate was 1.0 mL/min and injection volume was 10 ${\mu}l$. The chromatogram was monitored by photodiode array (PDA) detection at 280 nm for the identification of two flavonoid glycosides in P. trifoliata. The contents of the two components in P. trifoliata ranged from 0.32~13.02%.

Study of Rainfall-Runoff Variation by Grid Size and Critical Area (격자크기와 임계면적에 따른 홍수유출특성 변화)

  • Ahn, Seung-Seop;Lee, Jeung-Seok;Jung, Do-Joon;Han, Ho-Chul
    • Journal of Environmental Science International
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    • v.16 no.4
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    • pp.523-532
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    • 2007
  • This study utilized the 1/25,000 topographic map of the upper area from the Geum-ho watermark located at the middle of Geum-ho river from the National Geographic Information Institute. For the analysis, first, the influence of the size of critical area to the hydro topographic factors was examined changing grid size to $10m{\times}10m,\;30m{\times}30m\;and\;50m{\times}50m$, and the critical area for the formation of a river to $0.01km^2{\sim}0.50km^2$. It is known from the examination result of watershed morphology according to the grid size that the smaller grid size, the better resolution and accuracy. And it is found, from the analysis result of the degree of the river according to the minimum critical area for each grid size, that the grid size does not affect on the degree of the river, and the number of rivers with 2nd and higher degree does not show remarkable difference while there is big difference in the number of 1st degree rivers. From the results above, it is thought that the critical area of $0.15km^2{\sim}0.20km^2$ is appropriate for formation of a river being irrelevant to the grid size in extraction of hydro topographic parameters that are used in the runoff analysis model using topographic maps. Therefore, the GIUH model applied analysis results by use of the river level difference law proposed in this study for the explanation on the outflow response-changing characters according to the decision of a critical value of a minimum level difference river, showed that, since an ogival occurrence time and an ogival flow volume are very significant in a flood occurrence in case of not undertow facilities, the researcher could obtain a good result for the forecast of river outflow when considering a convenient application of the model and an easy acquisition of data, so it's judged that this model is proper as an algorism for the decision of a critical value of a river basin.

Study on Determination of Seven Transition Metal Ions in Water and Food by Microcolumn High-Performance Liquid Chromatography

  • Hu, Qiufen;Yang, Guangyu;Li, Haitao;Tai, Xi;Yin, Jiayuan
    • Bulletin of the Korean Chemical Society
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    • v.25 no.5
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    • pp.694-698
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    • 2004
  • A new method for the simultaneous determination of seven transition metal ions in water and food by microcolumn high-performance liquid chromatography has been developed. The lead, cadmium, mercury, nickel, cobalt, silver and tin ions were pre-column derivatized with tetra-(4-aminophenyl)-porphyrin ($T_4$-APP) to form the colored chelates which were then enriched by solid phase extraction with $C_{18}$ cartridge. The enrichment factor of 50 was achieved by eluted the retained chelates from the cartridge with tetrahydrofuran (THF). The chelates were separated on a ZORBAX Stable Bound microcolumn ($2.0{\times}50\;mm,\;1.8\;{\mu}m$)with methanol-tetrahydrofuran (95 : 5, v/v, containing 0.05 mol/L pyrrolidine-acetic acid buffer salt, pH = 10.0) as mobile phase at a flow rate of 0.5 mL/min and detected with a photodiode array detector from 350-600 nm. The seven chelates were separated completely within 2.0 min. The detection limits of lead, cadmium, mercury, nickel, cobalt, silver and tin are 4 ng/L, 3 ng/L, 6 ng/L, 5 ng/L, 5 ng/L, 6 ng/L, 4 ng/L respectively in the original samples. This method was applied to the determination of the seven transition metal in water and food samples with good results.

Determination of Boron by Ion Pair Liquid Chromatography with Chromotropic Acid (Chromotropic Acid를 착화제로 이용한 이온쌍 액체 크로마토그래피에 의한 붕소의 분리와 정량)

  • Yun, Young Ja;Yu, Gu Yong
    • Journal of the Korean Chemical Society
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    • v.39 no.4
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    • pp.288-293
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    • 1995
  • The separation and determination of boron with chromotropic acid (1,8-Dihydroxynaphthalene-3,6-disulfonic acid) as a complex agent has been studied using ion pair liquid chromatography. The use of tetrabutylammonium bromide added as an ion pair reagent to mobile phase (MeOH 61%, phosphate buffer 39% pH=8.5) allowed good separation of boron-chromotrophic acid complex anion and chromotrophic acid on poly(styrene-divinylbenzene) based reversed phase column (PRP-1, 15 $cm{\times}4.6$ mm i.d.). The complex formation between boric acid and chromotrophic acid was enhanced in the presence of 0.1 M tetrabutylammonium bromide, resulting in high sensitivity. The linear calibration was achieved over the boron concentration range of 0.5∼1000 ${\mu}g/L.$ The detection limit was 0.5 ${\mu}g/L$ (S/N=2). The proposed method was applied to the determination of boron in commercially available chemicals, $Na_2SO_4$, NaOH, KCl.

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Determination of residual novobiocin in livestock products and fisheries products by HPLC (HPLC를 이용한 축·수산 식품 중 잔류 노보비오신의 분석)

  • Lee, Byung Kyu;Lee, Cheol-Woo;Lee, Sang-Ju;Jung, Eun Ha;Lim, Hyun Kyun;Han, Sang Beom
    • Analytical Science and Technology
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    • v.20 no.4
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    • pp.347-354
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    • 2007
  • A simple and rapid high-performance liquid chromatography assay for the determination of residual novobiocin levels in bovine, porcine, chicken, flatfish and japanese eel muscle has been developed and validated. The separation condition for HPLC/UV was optimized with phenyl hexyl ($4.6{\times}150mm$, $5{\mu}m$) column with 10 mM monobasic sodium phosphate buffer (pH 2.5)/acetonitrile (50/50, v/v) as the mobile phase at a flow rate of 1.0 mL/min and detection wavelength was set at 254 nm. Residues were extracted from tissue by blending with methanol and lipid materials were removed with n-hexane. Then, the methanol extract was evaporated to dryness under a nitrogen stream, reconstituted in the mobile phase. Aliquot of the organic extract was decanted and filtered through $0.45{\mu}m$ syringe filter. The $20{\mu}L$ of the resulting solution was injected into the HPLC system. The calibration ranges were $0.5{\sim}5{\mu}g/g$ and calibration curves were linear with coefficients of correlation better than 0.95. The limits of quantification were $0.5{\mu}g/g$ for all muscles. The recoveries of bovine, porcine, chicken, flatfish and japaneseel muscles were 99.8%, 102.4%, 91.0%, 104.0% and 93.0%, respectively. The procedures were validated according to the CODEX guideline, determining specificity, linearity, accuracy, precision, quantitation limit and recovery.

Protein Binding Study of S-Ibuprofen Using High-Performance Frontal Analysis

  • Jin, Longmei;Choi, Du-Young;Liu, Haiyan;Row, Kyung-Ho
    • Bulletin of the Korean Chemical Society
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    • v.26 no.1
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    • pp.136-138
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    • 2005
  • High-performance frontal analysis (HPFA) was used for the determination of the binding constant of Sibuprofen to human serum albumin (HSA). This experiment was based on an Inertsil 100 Diol 5 column and sodium phosphate buffer (pH 7.4 and ionic strength of 0.17) as the mobile phase. The mixture of S-ibuprofen and HSA (70 $\mu$M) solution were directly injected into the HPFA column. An injection volume of 200 $\mu$L and a “estricted injection”method were applied to ensure the drug to be eluted as a zonal peak with a plateau. The unbound drug concentration was calculated from the peak height of the zonal peak. Scatchard analysis was used for evaluation of the binding constant (K) and binding affinity (nK) of S-ibuprofen to HSA, and the results were K = 2.833 ${\times}$ 10$^4$ [L mol$^{-1}$], nK = 4.935 ${\times}$ 10$^4$ [L mol$^{-1}$], respectively.

Effects of Force and Position Aspects of the Ankle Proprioception on the Balance Ability (발목관절에서 힘과 위치 측면의 고유수용성감각 수준이 균형능력에 미치는 영향)

  • Ree, Jae Sun;Hwang, Seonhong
    • Journal of Biomedical Engineering Research
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    • v.41 no.2
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    • pp.84-93
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    • 2020
  • Despite of a lot of studies about proprioception tests, there are little study results to investigate the relationship between the functional movement and proprioception level. In this study, we tried to perform quantitative analysis for the effect of ankle joint proprioception level on the one leg standing postural control ability. Nine healthy people volunteered for this study. Force and position aspects of proprioception were evaluated using the electromyography system (EMG) and mobile clinometer application, respectively. The center of pressure (COP) trajectories, measured by a pressure mat sensor, were used for quantitative analysis of balance for each subject. We computed indices and errors of force and position aspects of proprioception from the EMG and ankle angle. Mean velocity of total and anterior-posterior direction (Vm and Vm_ap), root mean squared distance in anterior-posterior direction (RDap), travel length (L), and area (A) of COP trajectories were also calculated as indices of postural control ability of subjects. Two aspects of proprioception showed the low correlation from each other as previous studies. However, the EMG error of gastrocnemius lateral activation showed a high correlation coefficient with COP variables such as Vm (ρ=0.817, p=0.007), Vm_ap (ρ=0.883, p=0.002), RDap (ρ=0.854, p=0.003), L (ρ=0.817, 0.007) and A (ρ=0.700, p=0.036). Within our knowledge, this is almost the first study that investigated the relationship between proprioception level and functional movement. These study results could support that the ankle joint proprioception facilitation exercise would have positive effects on functional balance rehabilitation interventions.

Qualitative and quantitative assessment of process related impurities in Brigatinib raw material and formulations using HPLC

  • Attada Tharun;Potnuru Jagadeesh;B Srinivasa Kumar;Kota Thirumala Prasad;Venkateswara Rao Anna
    • Analytical Science and Technology
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    • v.36 no.4
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    • pp.180-190
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    • 2023
  • The presence of process related impurities in any drug or the drug product was associated with its safety, stability and efficacy. The overall literature survey proved that there is no method published on the assessment of process related impurities in brigatinib. In this study, a simple, reliable and stable HPLC qualitative method was reported for quantification of process related impurities with easy and quick extraction procedure. The impurities along with standard brigatinib was resolved on Lichrospher® C18 (250 mm × 4.6 mm; 5 ㎛ particle size) column in room temperature using methanol, acetonitrile, pH 4.5 phosphate buffer in 55:25:20 (v/v) at 1.0 mL/min as mobile phase and UV detection at 261 nm. The method produces well resolved peaks at retention time of 4.60 min, 12.28 min, 3.37 min, 7.34 min and 8.39 min respectively for brigatinib, impurity A, B, C and D. The method produces a very sensitive detection limit of 0.0065 ㎍/mL, 0.0068 ㎍/mL, 0.0053 ㎍/mL and 0.0058 ㎍/mL for impurity A, B, C and D respectively with calibration curve linear in the concentration range of 22.5-135 ㎍/mL for brigatinib and 0.0225-0.135 ㎍/mL for impurities. The method produces all the validation parameters under the acceptable level and doesn't produces any considerable changes in peak area response while minor changes in the developed method conditions. The method can effectively resolve the unknown stress degradation products along with known impurities with less % degradation. The method can efficiently resolve and quantify the impurities in formulation and hence can suitable for the routine quality analysis of brigatinib in raw material and formulation.