Summary
The manipulation of electroosmotic flow in capillary electrophoresis was achieved by coating the inner wall of a fused silica capillary with the biopolymers α-chymotrypsinogen A and dextran sulfate. Simple coating procedures were based on flushing the fused silica capillary with α-chymotrypsinogen A solution to obtain a α-chymotrypsinogen A coating, or to additionally coat with dextran sulfate solution to obtain a α-chymotrypsinogen A-dextran sulfate coating. The biopolymers α-chymotrypsinogen A coated capillary exhibited strong reversed (anodic) electrosmotic flow values as high as −81.7×10−9 m2V−1s−1 at pH 2.0. The α-chymotrypsinogen A-dextran sulfate coated capillary exhibited a cathodic EOF of 62.2×10−9m2V−1 s−1 which remained virtually unaltered over the pH range 2–9. Both coatings showed high stability, as demonstrated by electroosmotic flow reproducibility of 1.0% and 0.7% RSD (n=50), respectively. The α-chymotrypsinogen A coating was found to be tolerant to 0.1 M HCl, whilst the α-chymotrypsinogen A-dextran sulfate coating was tolerant to 1 M NaOH, CH3OH and CH3CN. The coating-to-coating repeatability for the two coatings, as determined by the RSD of the resultant electroosmotic flow values, were 2.25% and 1.85% (n=4), respectively. Four anions and five cations were used as test substances to examine the separation performance of the α-chymotrypsinogen A and α-chymotrypsinogen A-dextran sulfate coatings and high efficiencies (80,000 to 200,000 theoretical plates) and rapid separations were obtained. The separation of isomers of chloroaniline was carried out using the α-chymotrypsinogen A-dextran sulfate coating and a pH 2.5 electrolyte in about one third of the time needed when using a FS capillary. A α-chymotrypsinogen A coated capillary was used for ultra-rapid separation of nitrate and nitrite at acidic pH using a co-electroosmotic flow mode. The separation was completed in less than 10s with a migration time reproducibility of 0.3% RSD (n=10) and sub-μM detection limits.
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Macka, M.; Johns, C.; Doble, P.; Haddad, P.R.LC-GC 2001,19, 38–47.
Macka, M.; Johns, C.; Doble, P.; Haddad, P.R.LC-GC 2001,19, 178–188.
Melanson, J.E.; Baryla, N.E.; Lucy, C.A.Anal. Chem. 2000,72, 4110–4114.
Horvath, J.; Dolnik, V.Electrophoresis,2001,22, 644–655.
Hjerten, S.J. Chromatogr. 1985,347, 191–198.
Cobb, K.A.; Dolnik, V.; Novotny, M.Anal. Chem. 1990,62, 2478–2483.
Xu, R.J.; Vidal-Madjar, C.; Sebille, B.; DiezMasa, J.C.J. Chromatogr. A,1996,730, 289–295.
Liu, Q.C.; Lin, F.M.; Hartwick, R.A.J. Liq. Chromatogr. 1997,20, 707–718.
Schmalzing, D.; Piggee, C.A.; Foret, F.; Carrilho, E.; Karger, B.L.J. Chromatogr. A 1993,652, 149–159.
Melanson, J.E.; Baryla, N.E.; Lucy, C.A.Trends in Anal. Chem. 2001,20, 365–374.
Tsuda, T.J. High Resolut. Chromatogr. Chromatogr. Commun. 1987,10, 622–624.
Jones, W.R.; Jandik, P.J. Chromatogr. 1991,546, 445–458.
Assi, K.A.; Altria, K.D.; Clark, B.J.J. Pharm. Biomed. Anal. 1997,15, 1041–1049.
Richards, M.P.J. Chromatogr. B 1994,657, 345–355.
Liu, Q.C.; Lin, F.M.; Hartwick, R.A.,J. Chromatogr. Sci. 1997,35, 126–130.
Wang, Y.; Dubin, P.L.Anal. Chem. 1999,71, 3463–3468.
Stathakis, C.; Arriaga, E.A.; Lewis, D.F.; Dovichi, N.J.J. Chromatogr. A 1998,817, 227–232.
Roche, M.E.; Anderson, M.A.; Pda, R.P.; Riggs, B.L.; Strausbauch, M.A.; Okazaki, R.; Wettstein, P.J.; Ianders, J.P.Anal. Biochem. 1998,258, 87–95.
Erim, F.B.; Cifuentes, A.; Poppe, H.; Kraak, J.C.J. Chromatogr. A 1995,708, 356–361.
Cordova, E.; Gao, J.; Whitesides, G.M.Anal. Chem. 1997,69, 1370–1379.
Li, M.X.; Liu, L.; Wu, J.T.; Lubman, D.M.Anal. Chem. 1997,69, 2451–2456.
Yao, Y.J.; Loh, K.C.; Chung, M.C.M.; Li, S.F.Y.Electrophoresis 1995,16, 647–653.
Chiu, R.W.; Jimenez, J.C.; Monnig, C.A.Anal. Chim. Acta 1995,307, 193–201.
Bossi, A.; Piletsky, S.A.; Turner, A.P.F.; Righetti, P.G.Electrophoresis 2002,23, 203–208.
Robb, C.S.; Yang, S.C.; Brown, P.R.Electrophoresis 2002,23, 1900–1905.
Wu, Q.R.; Lee, M.L.; Harrison, R.G.J. Chromatogr. A 2002,954, 247–258.
Gao, Q.F.; Yeung, E.S.Anal. Chem. 1998,70, 1382–1388.
Kleemiss, M.H.; Gilges, M.; Schomburg, G.Electrophoresis 1993,14, 515–522.
Nicole, E.B.; Lucy, C.A.J. Chromatogr. A 2002,956, 271–277.
Cunliffe, J.M.; Baryla, N.E.; Lucy, C.A.Anal. Chem. 2002,74, 776–783.
Katayama, H.; Ishihama, Y.; Asakawa, N.Anal. Chem. 1998,70, 2254–2260.
Katayama, H.; Ishihama, Y.; Asakawa, N.Anal. Chem. 1998,70, 5272–5277.
Bendahl, L.; Hansen, S.; Gammelgaard, B.Electrophoresis 2001,22, 2565–2573.
Rodriguez-Delgado, M.A.; Garcia-Montelongo, F.J.; Cifuentes, A.Anal. Chem. 2002,74, 257–260.
Varghese, J.; Cole, R.B.J Chromatogr. A 1993,652, 369–376.
Haginaka, J.J. Chromatogr. A 2000,875, 235–254.
Van Tassel, P.R.; Miras, D.; Hagege, A.; Voegel, J.C.; Schaff, P.J. Colloid and Interface Sci. 1996,183, 269–273.
Janssens, J.; Chevigne, R.; Louis, P.US Patent 5, 611, 903.
Carchon, H.; Eggermont, E.Electrophoresis 1982,3, 263–274.
Altria, K.D.; Kelly, M.A.; Clark, B.J.Chromatographia 1996,43, 153–158.
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Yang, W.C., Macka, M. & Haddad, P.R. Biopolymer-coated fused silica capillaries for high magnitude cathodic or anodic electro-osmotic flows in capillary electrophoresis. Chromatographia 57 (Suppl 1), S187–S193 (2003). https://doi.org/10.1007/BF02492101
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DOI: https://doi.org/10.1007/BF02492101