L‐Cysteine‐Bonded Polymeric Monolithic Stationary Phase for Enantioseparation of Dansyl Amino Acids in Capillary Liquid Chromatography

A chiral monolith stationary phase was fabricated by modifying the monolith surface using L-cysteine through a thiol-epoxy click reaction. L-cysteine-bonded polymer monolith was characterized by scanning electron microscopy/energy-dispersive X-ray and attenuated total reflectance Fourier-transformed infrared. The monomer content and modification temperature were carefully optimized to create a polymer monolith with excellent mechanical stability and permeability. Our findings revealed that the column morphology depended significantly on the porogen concentration and modification temperature for its morphology and efficiency. Adequate pores and binding sites were formed with the optimal porogen content, while a higher modification temperature improved the modification yield, enhancing peak shapes and increasing separation efficiency. The column demonstrated its capability for enantioseparation of dansyl glutamic acid, dansyl aspartic acid, dansyl methionine, and dansyl phenylalanine using a 60 mM ammonium acetate buffer solution and acetonitrile in a 20:80 v/v ratio. It maintained good mechanical stability and repeatability with no relative standard deviation exceeding 7%. These results indicated that the L-cysteine-bonded polymer monolith has excellent potential as a chiral stationary phase.