TY - JOUR
T1 - Non-bonded poly(ethylene oxide) polymer-coated column for protein separation by capillary electrophoresis
AU - Iki, Nobuhiko
AU - Yeung, Edward S.
N1 - Funding Information:
The Ames Laboratory is operated for the U.S. Department of Energy by Iowa State University under Contract No. W-7405Eng-82. This work was supported by the Director of Energy Research, Office of Basic Energy Sciences, Division of Chemical Sciences, and the Office of Health and Environmental Research. N.I. acknowledges support from a research fellowship of the Japan Society for the Promotion of Science.
PY - 1996/4/19
Y1 - 1996/4/19
N2 - A simple method to coat a non-ionic hydrophilic polymer onto the inner wall of a bare fused-silica capillary is established. The capillary is first filled with 1 M HCl solution, then flushed with 0.2% poly(ethylene oxide) solution containing 0.1 M HCl, and finally rinsed with the electrophoretic buffer. Each step takes only 5 min. For regeneration of the coating, this procedure is repeated. Four basic proteins were separated by this method around pH 3-7 with phosphate buffer. The separation efficiency in terms of peak shape was excellent compared to bare fused-silica capillaries and comparable to many other more sophisticated column-treatment methods. The reproducibility of migration times of proteins was less than 3% R.S.D. at pH 6 by manual operation of the coating process. Titration of the fused-silica surface with acid and the structure of poly(ethylene oxide) are important factors to form a stable coating via hydrogen bonding to the surface silanol groups.
AB - A simple method to coat a non-ionic hydrophilic polymer onto the inner wall of a bare fused-silica capillary is established. The capillary is first filled with 1 M HCl solution, then flushed with 0.2% poly(ethylene oxide) solution containing 0.1 M HCl, and finally rinsed with the electrophoretic buffer. Each step takes only 5 min. For regeneration of the coating, this procedure is repeated. Four basic proteins were separated by this method around pH 3-7 with phosphate buffer. The separation efficiency in terms of peak shape was excellent compared to bare fused-silica capillaries and comparable to many other more sophisticated column-treatment methods. The reproducibility of migration times of proteins was less than 3% R.S.D. at pH 6 by manual operation of the coating process. Titration of the fused-silica surface with acid and the structure of poly(ethylene oxide) are important factors to form a stable coating via hydrogen bonding to the surface silanol groups.
KW - Capillary columns
KW - Poly(ethylene oxide)-coated columns
KW - Proteins
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U2 - 10.1016/0021-9673(95)01158-7
DO - 10.1016/0021-9673(95)01158-7
M3 - Article
AN - SCOPUS:0029988347
VL - 731
SP - 273
EP - 282
JO - Journal of Chromatography A
JF - Journal of Chromatography A
SN - 0021-9673
IS - 1-2
ER -