Preparation of Affinity Column Based on Zr⁴⁺ Ion for Phosphoproteins Isolation

This paper has described about preparation of Zr⁴⁺ affinity column based on the poly(styreneco- glycidyl methacrylate) prepared by emulsion polymerization of styrene and glycidyl methacrylate in order to isolate phosphopeptide. The Zr⁴⁺ ions were introduced after the phophonation of an epoxy group on polymeric microspheres. The successful preparation of Zr⁴⁺-immobilized polymeric microsphere stationary phase was confirmed through Fourier transform infrared spectra, optical microscopy, scanning electron microscopy, X-ray photoelectron spectra and inductively coupled plasma-atomic emission spectrometer. The separation efficiency for Zr⁴⁺ affinity column prepared by slurry packing was tested to phosphonated casein and dephosphonated casein. The resolution time (min) of the phosphonated casein was higher than that of dephosphated casein for Zr⁴⁺ affinity polymeric microsphere by liquid chromatography. This Zr⁴⁺ affinity column can be used for isolation of phosphonated casein from casein using liquid chromatography