Separation of Tryptophan Enantiomers by using Chirobiotic® T HPLC Column

Chirobiotic® T HPLC 컬럼을 이용한 Tryptophan 이성질체 분리

  • Song, Sung-Moon (Department of Chemical Engineering, Chungnam National University) ;
  • Rang, Moon Jung (Department of Molecular Science, Paichai University) ;
  • Kim, In Ho (Department of Chemical Engineering, Chungnam National University)
  • 송성문 (충남대학교 화학공학과) ;
  • 랑문정 (배재대학교 분자과학과) ;
  • 김인호 (충남대학교 화학공학과)
  • Received : 2010.01.13
  • Accepted : 2010.06.22
  • Published : 2010.08.31

Abstract

D,L-tryptophans were separated by using $Chirobiotic^{(R)}$ T HPLC column. Mobile phases were the mixture of methanol and water(70:30, 80:20, 90:10, v/v). Experimental temperatures were adjusted as 25, 40 and $55^{\circ}C$ in order to compare retention times. Difference in D,L-tryptophan retention times was studied in terms of the interaction between stationary phase and tryptophans. Selectivity, resolution and efficiency of column were utilized to find an optimum separation condition. Retention times were shortened by increasing the amount of methanol in mobile phase and the temperature of column. The best selectivity and resolution was obtained with the temperature($25^{\circ}C$) and the ratio of mobilephase(70/30 v/v%).

$Chirobiotic^{(R)}$ T HPLC 컬럼을 이용하여 D, L-트립토판을 분리하였다. 이동상으로 메탄올과 증류수를 각각 70/30, 80/20, 90/10의 비율로 사용하고 체류시간을 비교하기 위해 온도를 25, 40, $55^{\circ}C$로 변화시켰다. 이동상 조성비를 변화시켜 D-트립토판과 L-트립토판의 체류시간 차이를 조사하였다. 선택도와 분리도, 컬럼의 효율성을 체류시간 관점에서 계산하여 최적의 분리 조건을 찾는 실험을 하였다. 이동상에서 메탄올의 양이 감소함에 따라 트립토판 체류시간이 짧아지고, 컬럼의 온도가 증가함에 따라 체류시간도 감소한다. 메탄올과 증류수의 비가 70/30일 때와 온도가 $25^{\circ}C$일 때 가장 좋은 선택도와 분리도를 얻었다.

Keywords

References

  1. Hyun, M. H., "Separation of Optical Isomers by LC," Minumsa, Seoul(1996).
  2. Nakamura, H., "Chiral Separations by HPLC," Hirokawa Publishing Co., Tokyo, Japan(1997).
  3. HPLC columns, Sigma-Aldrich, www.sigmaldrich.com.
  4. Jegal, J., Kim, J. H., Kim, J. H., Lee, K. H. and Lee, Y. T., "Enantioselective Membrane Based on Chitosan for the Separation of D- and L-Tryphophan," Kor. Mem. J., 5, 25-30(2003).
  5. Kim, K. I., "Study on Chiral Separation of Tryphophan by Preparative Liquid Chromatography with BSA-Silica Stationary Phase," MS thesis, Myungji University(2001).
  6. Park, M. B. and Kim, I. H., "Effect of Temperature and Eluent Composition on the Separation of Ketoprofen and Ibuprofen Racemates in Kromasil HPLC Column," Korean Chem. Eng. Res., 47, 54-58(2009).
  7. Kenji, H., Reiko, N., Akiko, M., Ryuichi, K. and Kiyoshi, Z., "Sensitive High-performance Liquid Chromatographic Assay for d-Amino-acid Oxidase Activity in Mammalian Tissues using a Fluorescent Non-natural Substrate, 5-Fluoro-d-Tryptophan," J. Chromatogr. A, 1106, 159-164(2003).
  8. Guisbert, A. L., Sharp, S., Petersen, J. A. and Risley, D. S., "Enantiomeric Separation of an AMPA antagonist using a Chirobiotic T column with HPLC and Evaporative Light-Scattering Detection," J. Liq. Chrom. Relat. Tech., 23(7), 1019-1028(2000). https://doi.org/10.1081/JLC-100101504
  9. Randon, J., Garnier, F., Rocca, J. L. and Maisterrena, B., "Optimization of the Enantiomeric Separation of Tryptophan Analogs by Membrane Processes," J. Mem. Sci., 175, 111-117(2000). https://doi.org/10.1016/S0376-7388(00)00402-6