Micellular Electrokinetic Capillary Chromatography in the Undergraduate Curriculum:: Separation and Identification of the Amino Acid Residues in an Unknown Dipeptide Using FMOC Derivatization

• Timothy G. Strein ^[1] ; James L. Poechmann ^[1] ; Mark Prudenti ^[1]
  1. [1] Bucknell University

     Bucknell University

     Borough of Lewisburg, Estados Unidos

     
• Localización: Journal of chemical education, ISSN 0021-9584, Vol. 76, Nº 6 (June), 1999, págs. 820-820
• Idioma: inglés
• Texto completo no disponible (Saber más ...)
• Resumen

  • This manuscript describes our efforts to introduce biochemistry students to micellar electrokinetic capillary chromatography (MEKC), a mode of capillary electrophoresis that employs micelles in the operating buffer. Unlike free solution capillary electrophoresis, MEKC is capable of resolving both charged and uncharged analytes because the micellar pseudo stationary phase allows for the separation of uncharged species. The experiment described herein includes a comparison of MEKC, employing sodium dodecyl sulfate (SDS) as the micelle-forming species, with reverse-phase HPLC. Both methods are used to determine the amino acid residues in an unknown dipeptide. Advanced undergraduate chemistry, biochemistry, and biology majors perform this experiment in the Biochemical Methods course at Bucknell University. The students cleave the peptide bond, derivatize the resultant amino acids with 9-fluorenylmethyl chloroformate (FMOC), and separate the FMOC–amino acid derivatives using HPLC and MEKC. This manuscript details the analytical procedures for the MEKC separation and presents typical student data obtained using this relatively new method.