Abstract

The separation of tryptophan enantiomers was carried out with medium-pressure liquid chromatography using BSA (bovine serum albumin)-bonded silica as a chiral stationary phase. The influence of various experimental factors such as pH and ionic strength of mobile phase, separation temperature, and the presence of organic additives on the resolution was studied. In order to expand this system to preparative scale, the loadability of sample and the stability of stationary phase for repeated use were also examined. The separation of tryptophan enantiomers was successful with this system. The data indicated that a higher separation factor (a) was obtained at a higher pH and lower temperature and ionic strength in mobile phase. Addition of organic additives (acetonitrile and 2-propanol) in mobile phase contributed to reduce the retention time of L-tryptophan. About $30\%$ of the separation factor was reduced after 80 days of repeated use.

주제어

#BSA-silica   #chiral separation   #chiral stationary phase   #liquid chromatography   #tryp-tophan enantiomers  

참고문헌 (20)

1. Commercially available brush-type chiral selectors for direct resolution of enantiomers , Perrin, S. P.;W. H. Pirkle , ACS Symp. Ser. / v.471,pp.43-66, 1991
   
2. Resolution of race-mic drugs on a new chiral column based on silica-immo-bilized cellobiohydrolase , Hermansson, J.;A. Grahn , J. Chromatogr. / v.A687,pp.45-59, 1994
   
3. Synthesis and chromatographic properties of an HPLC stationary phase based upon human serum albumin , Domenici, E.;C. Bertucci;P. Salvadori;G. Felix;I. Cahabne;S. Motellier;I. Wainer , Chromatographia / v.29,pp.170-176, 1990
   
4. High-performance liquid affinity chromatography , Larsson, P. O. , Methods Enzymol / v.104,pp.212-223, 1984
   
5. Protein column-based chiral reversed-phase liquid chromatography , Allenmark, S. G. , ACS Symp. Ser. / v.471,pp.114-125, 1991
   
6. A quantitative relationship between capacity factor and selector concentration in capillary electrophoresis and high performance liquid chromatography , Lloyd, D. K. , Electrophoresis / v.18,pp.958-964, 1997
   
7. Enantioselectivety of complex formation in ligand-exchange chromatographic systems with chiral stationary phase and/or chiral mobile phase , Davankov, V. A.;A. Kurganov;T. M. Ponomareva , J. Chromatogr / v.452,pp.309-316, 1988
   
8. Chromatographic optical resolution on polysaccharide carbamate phases , Okamoto, Y.;Y. Kaida;R. Aburatani;K. Hatada , ACS Symp. Ser. / v.471,pp.101-113, 1991
   
9. A chiral separation: An overview , Ahuja, S. , ACS Symp. Ser. / v.471,pp.1-26, 1991
   
10. Direct liquid chromatographic resolution of racemic drugs using α₁-acid glycoprotein as the chiral stationary phase , Hermansson, J. , J. Chromatogr. / v.269,pp.71-80, 1983
    
11. Direct liquid chromatographic separation of enantiomers on immobilozed protein stationary phases , Allenmark, S.;B. Bomgren;H. Boren , J. Chromatogr. / v.264,pp.63-68, 1983
    
12. Direct analytical and preparative resolution of enantiomers using albumin adsorbed to silica as a stationary phase , Erlandsson, P.;L. Hansson;R. Isaksson , J. Chromatogr. / v.370,pp.470-483, 1986
    
13. Retention data from affinity high-performance liquid chromatography in view of chemometrices , Kaliszan, R. , J. Chromatogr. / v.B715,pp.229-244, 1998
    
14. Protein chiral selectors in free-solution capillary electrophoresis and packed-capillary electrochromatography , Lloyd, D. K.;S. Li;P. Ryan , J. Chromatogr. / v.A694,pp.285-296, 1995
    
15. Chromatography on chiral stationary phase , Allenmark, S.;V. Schrig , J. Mater. Chem. / v.7,pp.1955-1963, 1997
    
16. Enantiomeric impurities in chiral catalysts, auxiliaries and synthons used in enantioselective synthesis , Armstrong, D. W.;J. T. Lee;L. W. Chang , Tetrahedron: Asymmetry / v.9,pp.2043-2064, 1998
    
17. Development of dihydrazide-activated silica supports for high-performance affinity chromatography , Ruhn, P. F.;S. Garver;D. S. Hage , J. Chromatogr. / v.A669,pp.9-19, 1994
    
18. Preparative chromatographic separation of enantiomers , Francotte, E.;A. Junker-Buchheit , J. Chromatogr. / v.576,pp.1-45, 1992
    
19. Separation procedures used to reveal and follow drug-protein binding , Sebille, B.;R. Zini;C. V. Madjar;N. Thuaud;J. P. Tillement , J. Chromatogr. / v.531,pp.51-77, 1990
    
20. Chiral separations in capillary electro-phoresis using proteins as stereoselective binding agents , Hage, D. , Electrophoresis / v.18,pp.2311-2321, 1997
    

이 논문을 인용한 문헌 (0)

1. 이 논문을 인용한 문헌 없음