Capillary electrochromatographic separation of proteins on a column coated with titanium dioxide nanoparticles.
A TiO2 nanoparticle (TiO2 NP)-coated open-tubular column for the capillary electrochromatographic separation of proteins is described. The surface chemistry of the TiO2 NPs on the inner wall of the fused silica was significantly affected by the running buffer. By varying of the phosphate buffer pH, only cathodic EOF was indicated. The results showed that TiO2 NPs are existed as a complexed form with the buffer ligand. Good separation of conalbumin (ConA), apo-transferrin (apoTf), ovalbumin (OVA), and BSA could be achieved with phosphate buffer (40 mM, pH 8.0) and an applied voltage of 15 kV. Five peaks of glycoisoforms of OVA were observed under these conditions. In comparison with the retention behavior of the analytes on the bare fused-silica column, the new column's high resolving power seems to be predominantly derived from the ligand exchange of the analytes with the phosphate adsorbed onto the TiO2 NPs. The method was also used to separate egg-white proteins. Both acidic and basic proteins in egg white were separated in a single run. The microheterogeneities of OVA could also be found in it. The separation efficiency for the main peak of OVA in egg white was around 10,000 plates/m.
Hsieh
Yi-Ling
YL
Department of Chemistry, National Taiwan University, Taipei, Taiwan.
Chen
Tse-Hsien
TH
Liu
Chuen-Ying
CY
D000818:Animals; D053801:Capillary Electrochromatography / Q000295:instrumentation* / Q000379:methods*; D002417:Cattle; D006031:Glycosylation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D053758:Nanoparticles / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D014025:Titanium / Q000737:chemistry*
D011506:Proteins; C009495:titanium dioxide; D014025:Titanium