• Title/Summary/Keyword: Chiral column

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Effect of acid or base additive in the mobile phase on enantiomer separation of amino alcohols using polysaccharide derived chiral columns (다당유도체를 기초로 한 키랄 컬럼에서 이동상에서의 산 또는 염기 첨가제가 아미노 알코올의 광학분리에 미치는 영향)

  • Huang, Hu;Jin, Jing Yu;Lee, Wonjae
    • Analytical Science and Technology
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    • v.22 no.4
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    • pp.313-318
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    • 2009
  • Liquid chromatographic enantiomer separation of amino alcohols was performed on several chiral columns based on polysaccharide derivatives under the mobile phase conditions containing acid or base additive. The chromatographic parameters were greatly influenced by the nature of the mobile phase containing acid or base additive as well as the used chiral columns. Compared to chromatographic results obtained in the mobile phase containing base additive (0.1% triethylamine), especially, Chiralcel OD showed dramatically enhanced separation factors and resolution factors with reduced capacity factors under the mobile phase condition containing acidic additive (0.1% trifluoroacetic acid). When the mobile phase containing 0.05% trifluoroacetic acid and 0.05% triethylamine was used on Chiralcel OD, the greatest separation factors and resolution factors among all other mobile phase conditions in this study were observed.

Chiral Separation of Salbutamol Enantiomers in Human Plasma

  • Seo, Joung-Min;Kim, Kyeong-Ho
    • Archives of Pharmacal Research
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    • v.17 no.4
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    • pp.244-248
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    • 1994
  • A stereoselective and sensitive high performance liquid chromatography using fluoresecence deterctor was examined for the determination of R(-) and S(+)-salbutamol in human plasma. Solid phase extraction method using silica as sorbent was used to extract salbutamol racemates from the plasma matrices. After fractionation and freeze-drying of the eluates containing salbutamol racemates, they were separated and quantified on a chirla stationary column. The detection limit of each enantiomer was 2 ng/ml in human plasma (S/N=3).

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Multidimensional Gas Chromatography-A Powerful Tool for the Analysis of Multicomponent Mixtures

  • Kim, Kyoung-Su
    • Preventive Nutrition and Food Science
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    • v.1 no.1
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    • pp.127-133
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    • 1996
  • The development of high resolution capillary columns and a large variety of different detectors led to a strong position of gas chromatography in instrumental analysis. Every effort has been made to solve sophisticated separation problems by column switching. Nowadays, several systems are commercially available for this purpose. The principle and the capabilities of multidimensional gas chromatography(MDGC) are illustrated by different applications in the field of modern flavor and essential oil research.

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Preparative Chromatographic Separaction: Simulated Moving Bed and Modified Chromatography Methods

  • Yi Xie;Koo, Yoon-Mo;Nien-Hwa Linda Wang
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.6
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    • pp.363-375
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    • 2001
  • Chromatography has been method of choice for the separation complex biologi-cal mixtures fro analytical purpose, particularly for the last fifty years. Its use has recently been extended to preparative separation where the productivity relative to the amount of resin and sol-vent used is a matter of concern. To overcome the inherent thermodynamic inefficiency of batch chromatography, as exemplified by the partial temporal usage of the resin and dilution of the product with the solvent, chromatography has been continually modified by separation engineers. Column switching and recycling represnet some of the process modifications that have brought high productivity to chromatography. Recently, the simulated moving bed (SMB) method, which claims a high separation efficiency based on counter-current moving bed chromatography. has be-come the mainstay of preparative separation, especially in chiral separation. Accordingly, this pa-per reviews the current status of SMB along with several chromatographic modification, which may be helpful in routine laboratory and industrial chromatographic practices.

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Development of Optically Active Chelate Resin for Direct Resolution of Enantiomers(III)-Synthesis of Copper(II) L-Proline Chelate Resin and Resolution of Enantiomers- (Enantiomer의 분리에 이용될 수 있는 Chelate Resin의 개발(제 3보)-Copper(II) L-Proline Chelate Resin의 제조 및 Enantiomer 분리-)

  • Kim, Kil-Soo;Jeon, Dong-Won
    • Journal of Pharmaceutical Investigation
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    • v.19 no.3
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    • pp.117-121
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    • 1989
  • A metal complex, copper (II) L-proline was chemically bound to ethylene glycol dimethacrylate and divinylbenzene crosslinked chloromethylated polystyrene and they were used as chiral chelate resin matrix for column chromatography to resolve enantiomers of DL-amino acids. The L-enantiomers eluted first and the degree of resolution on the polymer crosslinked with ethylene glycol dimethacrylate was superior to the polymer crosslinked with divinylbenzene.

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A Novel Design of Simulated Moving Bed (SMB) Chromatography for Separation of Ketoprofen Enantiomer

  • Yoon, Tae-Ho;Chung, Bong-Hyun;Kim, In-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.4
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    • pp.285-291
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    • 2004
  • A simulated moving bed (SMB) chromatography system is a powerful tool for preparative scale separation, which can be applied to the separation of chiral compound. We have de-signed our own lab-scale SMB chromatography using 5 HPLC pumps, 6 stainless steel columns and 4 multi-position valves, to separate a racemic mixture of ketoprofen in to its enantiomers. Our design has the characteristics of the low cost for assembly for the SMB chromatography and easy repair of the unit, which differs from the designs suggested by other investigators. It is possible for the flow path through each column to be independently changed by computer control, using 4 multi-position rotary valves and 5 HPLC solvent delivery pumps. In order to prove the operability of our SMB system, attempts were made to separate the (S)-ketoprofen enantiomer from a ketoprofen racemic mixture. The operating parameters of the SMB chromatography were calculated for ketoprofen separation from a batch chromatography experiment as well as by the triangle theory. With a feed concentration of 1 mg/mL, (S)-ketoprofen was obtained with a purity of 96% under the calculated operating conditions.

Production of (R)-Ethyl-4-Chloro-3-Hydroxybutanoate Using Saccharomyces cerevisiae YOL151W Reductase Immobilized onto Magnetic Microparticles

  • Choo, Jin Woo;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1810-1818
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    • 2015
  • For the synthesis of various pharmaceuticals, chiral alcohols are useful intermediates. Among them, (R)-ethyl-4-chloro-3-hydroxybutanoate ((R)-ECHB) is an important building block for the synthesis of L-carnitine. (R)-ECHB is produced from ethyl-4-chloro-3-oxobutanoate (ECOB) by a reductase-mediated, enantioselective reduction reaction. The Saccharomyces cerevisiae YOL151W reductase that is expressed in Escherichia coli cells exhibited an enantioselective reduction reaction toward ECOB. By virtue of the C-terminal His-tag, the YOL151W reductase was purified from the cell-free extract using Ni2+-NTA column chromatography and immobilized onto Ni2+-magnetic microparticles. The physical properties of the immobilized reductase (Imm-Red) were measured using electron microscopy, a magnetic property measurement system, and a zeta potential system; the average size of the particles was approximately 1 μm and the saturated magnetic value was 31.76 emu/g. A neodymium magnet was used to recover the immobilized enzyme within 2 min. The Imm-Red showed an optimum temperature at 45℃ and an optimum pH at 6.0. In addition, Bacillus megaterium glucose dehydrogenase (GDH) was produced in the E. coli cells and was used in the coupling reaction to regenerate the NADPH cofactor. The reduction/oxidation coupling reaction composed of the Imm-Red and GDH converted 20 mM ECOB exclusively into (R)-ECHB with an e.e.p value of 98%.

Enantioselective Bioconversion Using Escherichia coli Cells Expressing Saccharomyces cerevisiae Reductase and Bacillus subtilis Glucose Dehydrogenase

  • Park, Hyun-Joo;Jung, Ji-Hye;Choi, Hye-Jeong;Uhm, Ki-Nam;Kim, Hyung-Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.20 no.9
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    • pp.1300-1306
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    • 2010
  • Ethyl (R, S)-4-chloro-3-hydroxybutanoate (ECHB) is a useful chiral building block for the synthesis of L-carnitine and hypercholesterolemia drugs. The yeast reductase, YOL151W (GenBank locus tag), exhibits an enantioselective reduction activity, converting ethyl-4-chlorooxobutanoate (ECOB) exclusively into (R)-ECHB. YOL151W was generated in Escherichia coli cells and purified via Ni-NTA and desalting column chromatography. It evidenced an optimum temperature of $45^{\circ}C$ and an optimum pH of 6.5-7.5. Bacillus subtilis glucose dehydrogenase (GDH) was also expressed in Escherichia coli, and was used for the recycling of NADPH, required for the reduction reaction. Thereafter, Escherichia coli cells co-expressing YOL151W and GDH were constructed. After permeablization treatment, the Escherichia coli whole cells were utilized for ECHB synthesis. Through the use of this system, the 30 mM ECOB substrate could be converted to (R)-ECHB.

A Newly Developed Analytical and Semi-preparative Enantiomer Separation of Fluoxetine using Polysaccharide-derived Chiral Stationary Phases by High Performance Liquid Chromatography (고성능 액체 크로마토그래피에 의한 다당 유도체의 키랄 고정상에서 플록세틴의 새롭게 개발된 분석 및 반분취의 광학분리)

  • Kim, Seok Jin;Nam, Kyung Wook;Park, Bohyun;Islam, Md. Fokhrul;Lee, Wonjae
    • KSBB Journal
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    • v.31 no.3
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    • pp.186-191
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    • 2016
  • A liquid chromatographic method for the enantiomer separation of fluoxetine was performed using covalently bonded and coated type polysaccharide-derived chiral stationary phases (CSPs). The degree of enantioseparation is affected by the used CSPs and mobile phases. The performance of Chiralpak IC was superior to the other CSPs used in this study. Out of various solvent composition and additives, the greatest separation and resolution was observed using Chiralpak IC with mobile phase of 2-propanol in hexane with diethylamine as an additive. Semi-preparative separation of fluoxetine was performed on the analytical Chiralpak IC column to obtain (R)- and (S)-fluoxetine enantiomer with high chemical and optical purity. From the overall study, the developed liquid chromatographic method on polysaccharide-derived CSPs is expected to be very useful for the enantiomer separation of fluoxetine.