Rapid Chiral Separation by Flow-Through Chromatography with a Biporous Stationary Phase


Enantiomer separation is an area of increasing importance in chemistry and in the pharmaceutical industry. Although liquid chromatography is one of the most practical processes for chiral resolution, the high mass-transfer resistance of most commercially available packings makes chiral resolution time-consuming. In this work, a novel chiral stationary phase (CSP) with wide pores was prepared by coupling bovine serum albumin to a biporous resin with triazine as a linker. The rigid biporous medium was fabricated by radical suspension copolymerization with solid granules and solvents as porogenic agents. Studies by scanning electron microscopy and mercury intrusion porosimetry revealed the matrix contained two types of pore—micropores smaller than 180 nm and macropores of 500–7300 nm. Because the macropores provide convective flow channels for the mobile phase, the chromatographic process can be operated at high flow rate with low back-pressure. The biporous CSP was applied to the resolution of d, l-tryptophan (Trp). When the flow velocity was as high as 1800 cm h−1, d, l-Trp could still be separated. By comparison of the resolution of d, l-Trp on this CSP at high flow velocities with that predicted theoretically for conventional supports we concluded that the increased flow velocity had little effect on the resolution of enantiomers with biporous packings. The results indicate that the protein biporous CSP is promising for high-speed resolution of enantiomers.

This is a preview of subscription content, log in to check access.


  1. Lai XH, Ng SC (2004) J Chromatogr A 1031:135–142

    CAS  Google Scholar 

  2. Garcés J, Franco P, Oliveros L (2003) Tetrahedron Asymmetry 14:1179–1185

    Google Scholar 

  3. Hefnawy MM, Aboul-Enein HY (2004) J Pharm Biomed Anal 35:535–543

    CAS  Google Scholar 

  4. Lu Y, Li CX, Liu XH, Huang WQ (2002) J Chromatogr A 950:89–97

    CAS  Google Scholar 

  5. Park JH, Ryu JK, Park JK (2001) Chromatographia 53:405–408

    CAS  Google Scholar 

  6. Millot MC, Taleb NL, Sebille B (2002) J Chromatogr B 768:157–166

    CAS  Google Scholar 

  7. Sadakane Y, Matsunaga H, Nakagomi K (2002) Biochem Biophys Res Commun 295: 587–590

    CAS  Google Scholar 

  8. Hjertén S, Li YM, Liao JL, Mohammad J, Nakazato K, Pettersson G (1992) Nature 356:810

    Google Scholar 

  9. Lubda D, Lindner W (2004) J Chromatogr A 1036:135–143

    CAS  Google Scholar 

  10. Afeyan NB, Gordon NF, Mazsaroff I, Varady L, Fulton SP, Yang YB, Regnier FE (1990) J Chromatogr 519:1–29

    CAS  Google Scholar 

  11. Collins WE (1997) Sep Purif Methods 26:215–253

    Article  CAS  Google Scholar 

  12. García MC, Torre M (2000) J Chromatogr A 880:169–187

    Google Scholar 

  13. Gustavsson PE, Larsson PO (1996) J Chromatogr A 734:231–240

    CAS  Google Scholar 

  14. Palsson E, Smeds AL, Petersson A, Larsson PO (1999) J Chromatogr A 840:39–50

    CAS  Google Scholar 

  15. Shi Y, Sun Y (2003) Chromatographia 57:29–35

    CAS  Google Scholar 

Download references


The authors are grateful for the financial support provided by the Natural Science Foundation of China (grant No. 20276051).

Author information



Corresponding author

Correspondence to Yan Sun.

Additional information

Revised: 8 October 2004 and 3 January 2005

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Li, Y., Shi, QH. & Sun, Y. Rapid Chiral Separation by Flow-Through Chromatography with a Biporous Stationary Phase. Chroma 61, 213 (2005). https://doi.org/10.1365/s10337-005-0508-9

Download citation


  • Column liquid chromatography
  • Flow-through chromatography
  • Chiral stationary phase
  • Enantiomer resolution
  • Bovine serum albumin
  • d,l-Tryptophan