• Title/Summary/Keyword: gradient elution chromatography

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Prediction of Formic Acid Chromatogram in Gradient Elution Chromatography

  • Won, Hye-Jin;Kim, In-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.1
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    • pp.31-36
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    • 2001
  • Optimal operation in chromatography is needed to save operation time and the solvent used in multiple chromatographic runs. To this end, many simulation studies of chromatography process have been performed. The relationship between the distribution coefficient and the ionic strength is important in gradient elution ion chromatography. Experimental runs and computer simulations were carried out under linear gradient elution condition in order to compare the experiments and the simulation. Experiments were performed with formic acid under isocratic conditions to determine the simulation equation parameters. Computer simulation was based on three equations which related distribution with ionic strength as follows; K=${\alpha}$I(sup)-${\beta}$, K=A+BI+Cl$^2$and K=y(sub)0+A$_1$$.$e(sup)(-I/m$_1$). The effects of gradient slope on the chromatograms are discussed, and good agreement between the experimental and the simulated results is shown.

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Recent Developments in High-performance Liquid Chromatography of Lipids

  • Christie, William W.
    • Journal of the Korean Applied Science and Technology
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    • v.10 no.1
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    • pp.1-8
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    • 1993
  • The possibilities for HPLC analysis of lipids have been revolutionised by the availability of evaporative light-scattering detectors, with which the response is independent of the nature of the mobile phase and does not depend On the presence of specific chromophores in the lipids. It was thus possible to develop an HPLC procedure, involving ternary gradient elution, for separating all the lipid classes in animal tissues in a single step. Although reversed-phase HPLC has been widely used for the analysis of molecular species of lipids, sliver ion chromatography can be a valuable alternative. For example, a stable silver ion column for HPLC was developed which permitted resolution of molecular species of triacylglycerols, even from such complex samples as fish oils, again With light-scattering detection and gradient elution. The capacity for HPLC resolution of diastereomeric diacyl-sn-glycerol derivatives, prepared from triacylglycerols. has lead to a new simple method for stereospecific analysis of the latter.

HPLC Study for Egg White Analysis (달걀 단백질 분석을 위한 HPLC 연구)

  • Jeon, Young-Ju;Lee, Eun;Kim, In-Ho
    • KSBB Journal
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    • v.22 no.2
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    • pp.119-122
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    • 2007
  • Since egg white contains various protein, it is important to research the protein distribution of egg white. Specially, lysozyme and ovalbumin important proteins, are used in medicine and food industry. Reversed phase high performance liquid chromatography has been used for separation of egg white, and column of RP-HPLC is available in variety. We have used C4, C8 and C18 columns to obtain chromatograms by varying carbon chain length of stationary phase. Long carbon chain length of stationary phase has good separation of egg white. Also, we have changed the composition of mobile phase (acetonitrile, water, and trifluoroacetic acid) to find optimum chromatograms. Acetonitrile and water composition of 50 : 50 show many peaks from egg white. Isocartic and gradient elution in RP-HPLC were used to compare the chromatography of egg white.

Simultaneous Purification of Enterotoxin A and C by Fast Protein Liquid Chromatography (FPLC에 의한 Staphylococcal Enterotoxin A와 C의 동시분리)

  • Lee, Jung-Hee;Kim, Jong-Bae;Shin, Heuyn-Kil
    • Korean Journal of Food Science and Technology
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    • v.20 no.6
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    • pp.856-861
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    • 1988
  • A new method developed for simultaneous purification of enterotoxin A and C from Staphylococcus aureus strain L 350/1 consisted of chromatography on carboxymethyl (CM)-cellulose using a buffer of variable pH, gel filtration on Ultro gel, and fast protein liquid chromatography(FPLC) using a buffer of variable pH. The enterotoxin A and C were purified by three steps: batchwise adsorption from culture supernatant on Amberlite CG-50; chromatography on CM-cellulose using a buffer of constant pH and molarity; and gel filtration on Sephadex G-75. The purified enterotoxin appeared homogeneous by gel diffusion and polyacrylamide gel electrophoresis. Upon treatment with CM-cellulose using a elution of variable pH, enterotoxin A and C were so close that they were not separated completely. After elution from gels, the enterotoxins appeared as a single peak at the same position. Gel filtration gave a reaction of complete identity to enterotoxin A and C in Ouchterlony immunodiffusion. In FPLC using a CM-cellulose, enterotoxin A and C were simultaneously separated at pH 8.6 and 6.8. When each fraction was performed to gel immunodiffusion, at peak of enterotoxin A and C were not detected each other. In a method of elution by pH-gradient was to be more efficient as a simultaneous separation method in terms of speed, yields and simplicity. The purified toxin A and C were identical to type A and C reference enterotoxin on both disc electrophoresis and Ouchterlony gel diffusion.

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Purification of Eicosapentaenoic Acid (EPA) by Density Gradient Supercritical $CO_2$ Chromatography (밀도구배 초임계 $CO_2$ 크로마토그래피에 의한 Eicosapentaenoic Acid (EPA) 정제)

  • 유병삼;변상요
    • KSBB Journal
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    • v.14 no.1
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    • pp.109-113
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    • 1999
  • Supercritical $CO_2$ chromatography was applied for purification of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from fish oil. Various supercritical $CO_2$ pressures were tested to find out the pressure effects on solubility and selectivity of low fatty acids in the silver nitrate column. The solubility of low fatty acids was increased as the supercntical $CO_2$ Pressure increased. However, the selectiviy between low fatty acids and EPA waw decreased. Stepwise density gradient method was applied to increase the purification efficiency of EPA. Low fatty acids were easily separated at the early elution steps with low $CO_2$ densities. Successive fractions containing 92.1~97.8% of EPA were collected. The average concentration of three purified fractions was 95.6% with the recovery rate of 30%.

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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.

Separation Characteristics of Oligodeoxynucleotides by High-Performance Membrane Chromatography (고성능 막 크로마토그래피에 의한 Oligodeoxynucleotides의 분리특성)

  • Kim, Jung-Il;Hong, Seung-Bum;Sun, Hyang;Row, Kyung-Ho
    • KSBB Journal
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    • v.15 no.6
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    • pp.605-608
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    • 2000
  • Oligodeoxynucleotides (ODNs) were separated by high-performance membrane chromatography (HPMC), a combined system of chromatography and membrane. The separation mechanism involved anion-exchange, and the stationary phase was cation CIM (Convective Interaction Media) DEAE disk (16${\times}$3 mm). Two types of mobile phase were used, buffer A (20mM Tris-HCl, pH 7.4) and buffer B (buffer A + 1M NaCl). As the amount of NaCl dissolved in buffer linearly increased, the retention time shortened, which enabled a gradient elution mode. Based on the number of theoretical plates and resolution observed, the optimum mobile phase and operating condition (Buffer A/Buffer B=50/50 - 20/80 vol%, gradient time 2 min) were experimentally determined. In this experimental condition, ODNs were separated within 2 min at a mobile phase flow rate of 6 ml/min.

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A study of test method for impurities(related compounds) in pharmaceutical products

  • Ko, Yong-Seok;Jang, Seung-Jae;Kang, Chan-Soon;Choi, Bo-Kyung;Kim, Hye-Soo;Kim, Eun-Jung;Cho, Myoeng-Sin;Hong, Chong-Hui;Kim, Sang-Hyun
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.280.1-280.1
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    • 2003
  • The high-performance liquid chromatography method was performed for test method development of related compounds in pharmaceuticals. Using reverse-phase column and gradient elution of 1 % acetonitrile-acetonitrile: H20:triethylamine (70:30:0.5), lansoprazole, 2-hydroxybenzimidazole, 2-mercaptobenzimidazole, lansoprazole sulfone, lansoprazole sulfide could be individually identified and quantitated. (omitted)

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Rational and efficient approach to the preparation of the active fractions of Scutellaria baicalensis (황금(Scutellaria baicalensis) 유효분획물 제조의 합리적이고 효율적인 접근방법)

  • Kim, Doo-Young;Kim, Won Jun;Kim, Jung-Hee;Oh, Sei-Ryang;Ryu, Hyung Won
    • Journal of Applied Biological Chemistry
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    • v.62 no.1
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    • pp.31-38
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    • 2019
  • Scutellaria baicalensis Georgi (Scutellariae Radix) has been widely used as a dietary ingredient and traditional herbal medicine such as diuretic, hyperlipidemia, antibacterial, anti-allergy, anti-inflammatory and anticancer properties. In this study, the isolation of biomarkers or bioactive compounds from complex S. baicalensis extracts represents an essential step for de novo identification and bioactivity assessment. The bioactive fraction consisted of eight compounds which was chromatographed on an analytical high performance liquid chromatography column using two different gradient runs. A simulative replacement of the analytical column with a medium pressure liquid chromatography and open column allowed the determination of gradient profile to allow sufficient separation in the preparative scale. From the optimized method, eight standard compounds have been identified in the fractions. In addition, MS, UV, HRMS detection was provided by ultraperformance liquid chromatographyequadrupole time-of-flight mass spectrometry (UPLC-QTof-MS) of all fractions. Therefore, this scale up procedure was successfully applied to a S. baicalensis extract.