Zirconia non-hydrolytic sol-gel coated column for the capillary electrochromatographic separation of proteins
Date Issued
2007
Date
2007
Author(s)
Lin, Po-Ting
DOI
zh-TW
Abstract
A ZrO2 nanoparticle-coated open-tubular column was prepared through the non-hydrolysis sol-gel reaction of zirconium isopropoxide propanol complex and zirconium chloride with silanol groups of the fused-silica capillary. The condensation reaction was carried out by gradient temp. raising to 300oC to form a monolayer. EOF measurements and SEM image were used to monitor the completion of reactions. The property of ZrO2 on the inner wall of capillary was significantly affected by the running buffer. Cathodic EOF at pH between 4~6 was indicated. It can be elucidated that the negatively charge phosphate ion in the buffer absorb strongly on the surface of ZrO2 resulting in the formation of phosphate ions-capped ZrO2 NPs complex. We selected four proteins, such as conalbumin (ConA), apo-transferrin (apoTf), ovalbumin (OVA), and bovine serum albumin (BSA) as model compounds, whose molecular weights and pI values are alike to each other. They could be separated with phosphate buffer (10 mM, pH 8.0) with 5 % MeOH and an applied voltage of 20 kV. In addition, five peaks of glycoisoforms of OVA and two forms of BSA were observed under these conditions. The column was also used to separate egg-white proteins. In addition to separating OVA and ConA, it can resolve three microheterogeneities of OVA and two forms (iron-free and iron-saturated) of ConA. In comparison with the retention behavior of the analytes on the bare fused-silica column, the new column of high resolving power seems to be predominantly derived from the ligand exchange of the analytes with the phosphate adsorbed onto the ZrO2 .
Subjects
二氧化鋯,毛細管電層析,蛋白質
zirconia,CEC,protein
Type
thesis
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