• Title/Summary/Keyword: preparative-scale

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Large Scale Purification of KRF-001 on the Preparative HPLC (Preparative HPLC를 이용한 KRF-001의 대량분리정제)

  • 이항우;김무경정태숙복성해
    • KSBB Journal
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    • v.9 no.4
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    • pp.385-394
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    • 1994
  • Large scale purification to get antifungal antibiotic KRF-001 of 90% purity, was investigated using preparative HPLC. Crude KRF-001 was purified by XAD-7 adsorption chromatography, acid precipitation and microfiltration. Microfiltration was the most effective isolation method of crude KRF-001. The purification methods using C18 chromatography was convenient compared with the conventional methods. Delta PAK C18 column and Bonda PAK C18 column were adapted large scale purification of KRF-001. Gradient system of prep HPLC using Delta PAK C18 column was more effective. With these conditions, final recovery of KRF-001 yielded 77%.

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Isolation of ginsenosides Rb1, Rb2, Rc Rd, Re, Rf and Rg1 from cinseng root by high performance liquid chromatography

  • Paik, Nam-Ho;Park, Man-Ki;Choi, Kang-Ju;Cho, Yung-Hyun
    • Archives of Pharmacal Research
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    • v.5 no.1
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    • pp.7-12
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    • 1982
  • Ginsenosides Rb1, Rb2, Rc, Rd, Re, Rf and Rg1 were effectively isolated from ginseng root by preparative liquid chromatography (LC) on two PrepPAK-500/c18 cartridges in series and semipreparative LC on a .mu. Bondapak cabohydrate analysis column, a .mu.Bondapak C18 column or a .mu. Porasil column. The identities of the isolated ginsenosides were confirmed by analytical high-performance liquid chromatography (HPLC) and infrared spectrophotometry. By this method large scale isolation of pure ginsenosides was efficiently accomplished.

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Downstream Processing of Recombinant Hirudin Produced in Saccharomyces cerevisiae

  • Chung, Bong-Hyun;Kim, Won-Kyung;Rao, K.Jagannadha;Kim, Chul-Ho;Rhee, Sang-Ki
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.179-183
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    • 1999
  • A recombinant form of hirudin, a potent thrombin-specific inhibitor derived from the bloodsucking leech, was expressed as a secretory product in Saccharomyces cerevisiae under the control of GALl0 promoter and the mating factor $\alpha$pre-pro leader sequence. In an attempt to produce recombinant hirudin (r-Hir) of therapeutic purity in large quantities, the fed-batch fermentation was carried out by using this recombinant yeast, and subsequently downstream processing was developed with the preparative-scale column chromatography systems. About 234 mg/l of biologically active r-Hir was produced as a secretory product by the fed-batch fermentation strategy developed for an efficient downstream processing. Using a two-step chromatography process (an anion exchange chromatography followed by the reverse phase HPLC), the r-Hir was purified to>98% with an overall recovery yield of 84%. According to the N-terminal amino acid sequencing, the purified r-Hir was found to have the predicted N-terminal amino acid sequence. The biological activity of the purified r-Hir to inhibit thrombin was also identical to that of the commercial hirudin.

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Simulated Moving Bed [SMB] (모사 이동층 크로마토그래피)

  • Lee Chong-Ho;Koo Yoon-Mo
    • KSBB Journal
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    • v.20 no.3
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    • pp.192-196
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    • 2005
  • Chromatography has been a method of choice in the separation of complex biological mixtures for the analytical purpose in particular for the last half of century. In current years, chromatographic method extends its use to the preparative separation where the productivity per resin amount and solvent use become a matter of concern. Recently, simulated moving bed (SMB) method which claims high separation efficiency of the ideal counter-current moving bed chromatography has become a workhorse of preparative separation. SMB technology was developed in the early 1960s for large-scale hydrocarbon separation by UOP and approximately 120 Sorbex units have been licensed to date. Recently, SMB separation technology has been successfully extended from hydrocarbons and sugars to fine chemicals, particularly biochemicals, from laboratory to pilot to production plant. In this paper, the current status of SMB and its modifications were reviewed.

Characteristics of Preparative Liquid Chromatography (제조용 액체 크로마토그래피의 특성)

  • Row, Kyung-Ho;Jin, Yin-Zhe
    • KSBB Journal
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    • v.20 no.3
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    • pp.149-163
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    • 2005
  • Recently preparative liquid chromatography (PLC) has been used more frequently to separate drugs and natural substances. This modern separation methodologies require reliable tools that perform on a high level in terms of efficiency and reproducibility. However, large-scale PLC easily tends to reduce the yield and purity of the product. To promote the separation efficiency of PLC, we need to properly understand the controlling effects of the process, which may enable to predict the process and to improve the design and operation of PLC. Progress in computer technology allows the use of sophisticated models, provided their parameters can be measured. Some hardwares as well as softwares for PLC were already commercially available. In this work, the separation characteristic of PLC will be reviewed and compared on both the software and the hardware.

Recent Developments in Analytical and Preparative Scale Enantioseparations

  • Chankvetadze, Bezhan
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.161-162
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    • 2002
  • Field of enantioseparations represents one of the actual topics of chemistry. Significant, in some cases dramatic differences in a desired activity as well as in toxic properties of enantiomers of chiral drugs, food additives, agrochemicals, cosmetic products, etc. represents a driving force in this field. The enantiomers of all chiral bioactive compounds have to be isolated and to be tested. Besides the ethical or environmental reasons for developing single enantiomers, it may be a real therapeutic benefit and in some cases, it has been used as a strategy for extending patent life. (omitted)

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Centrifugal Liquid Chromatography with Application of the N-hexadecyl-L-proline Coated Reversed Phase for Separation of amino Acid Enantiomers (N-hexadecay-L-proline이 코팅된 역상 원심 액체크로마토그래피에 의한 아미노산 이성질체의 분리)

  • Sun Haing Lee;Tae Sub Oh;Hae Woon Lee
    • Journal of the Korean Chemical Society
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    • v.36 no.6
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    • pp.849-856
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    • 1992
  • We prepared a simple, economic and versatile preparative system for an enantiomeric separation. Hydrophobic amino acid enantiomers were resolved in the preparative scale by a centrifugal liquid chromatography on the N-hexadecyl-L-proline coated reversed phase. The factors controlling the retention and resolution of racemic amino acids such as the concentration of Cu(Ⅱ), pH of the eluent, the type and content of organic modifier, and rpm of CLC were examined. Several mg of hydrophobic amino acid enantiomers were separated preparatively. To separate all of different amino acid enantiomers, various coating material will be investigated.

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Effect of Sample-loading on Fractionation Efficiency (FE) in a Large Scale Splitter-less Gravitational SPLITT Fractionation (GSF)

  • Lee, Seung-Ho;Lee, Ji-Yeon;Lee, Tae-Woo;Jung, Euo-Chang;Cho, Sung-Kwang
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4291-4296
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    • 2011
  • Gravitational SPLITT fractionation (GSF) provides separation of colloidal particles into two subpopulations in a preparative scale. Conventionally, GSF is carried out in a thin rectangular channel having two inlets and two outlets at the top and bottom of the channel, respectively. And the channel is equipped with two flow-splitters, one between the top and bottom inlets and another between the top and bottom outlets. A large scale splitter-less GSF system had been developed, which was designed to operate in the full feed depletion (FFD) mode. In the FFD mode, there is only one inlet through which the sample is fed, thus preventing the sample dilution. In this study, the effect of the sample-loading (in the unit of g/hr) on the fractionation efficiency (FE, number% of particles in a GSF fraction that have the sizes expected by theory) of the new large scale splitter-less FFD-GSF system was investigated. The system was tested in the sample-loading range of 3.0-12.0 g/hr with polyurethane latex beads (PU) and sea-sediment. It was found that there is an optimum range in the sample-loading for a FFD-GSF separation. It was also found that there is a general tendency of FE decreasing as the concentration of the sample suspension increases.