Zirconia nanoparticles coated column for the CEC Separation of iron-binding- and phosphorylated-proteins
Date Issued
2009
Date
2009
Author(s)
LEE, CHIA-HSIN
Abstract
A ZrO2 nanoparticles(NPs) coated column was prepared through the sol-gel reaction of zirconium(IV) oxychloride with silanol groups of the fused-silica capillary. The condensation reaction was carried out at 350 oC for 8 h. Electroosmotic flow (EOF) measurements and SEM image were used for the characterization of the ZrO2 on the inner wall of capillary. Below the pI value (pH 5-6), cathodic EOF was elucidated that the phosphate buffer adsorb tightly on the zirconia surface resulting in a negative charge surface. In this work, iron-binding-, phosphorylated- and glyco-protein were selected as the model compounds. For the optimization, the effects of pH, concentration and buffer type as well as the organic modifier on the separation were studied. The results of iron-binding proteins show that the retention time is Mb > Hb. It is corresponding to the binding constants of ZrO2 NPs. α-, and β-subunit of Hb could be separated in borate buffer (20 mM, pH 9.0), MeOH (20 %, v/v). A greater affinity of α-casein and BSA toward the stationary phase was indicated by the comparison with that of ConA and Tf as pH increase. Interestingly, 14 peaks of glycoisoforms of OVA were observed under borate buffer (40 mM, pH 9.0). The established method was also applied to the determination of ConA and OVA in egg-white of chicken and duck egg. The concentration of ConA in chicken egg-white is 0.72 mM.
Subjects
zirconia nanoparticle
CEC
protein
Type
thesis
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