Development of Amine Capillary Column Applied to the Analysis of Basic Compounds by Electrochromatography
Affiliation(s)
1 Departamento de Tecnología Farmacéutica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina. 2 Consejo Nacional de Investigación Científica y Tecnológica (CONICET), Buenos Aires, Argentina.
1 Departamento de Tecnología Farmacéutica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina. 2 Consejo Nacional de Investigación Científica y Tecnológica (CONICET), Buenos Aires, Argentina.
ABSTRACT
An amine capillary column was developed in a fused-silica capillary as stationary phase and applied to separation of basic compounds by capillary electrochromatography (CEC). The functionalized capillary was prepared by inmobilization of 3-Aminopropyl)triethoxysilane (APTES). The CEC conditions including APTES and buffer concentration, pH and applied voltage were investigated to obtain the optimal CEC system for the separation of three anesthetics as basic test molecules. The capillary column provided an efficiency of up to 20,000 plates/m. Lidocaine, ketamine and xilacine were baseline separated under the running conditions with 10 mM Na2HPO4 pH 9.0 as BGE. The applied voltage was 5 kV temperature was set at 25oC and UV detection was performed. The resolutions were 4.97 and 1.53 for ketamine, lidocaine and xilacine, respectively. The column used in CEC mode showed better separation of the anesthetics compared with those used in the capillary zone mode. The comparison with reversed stationary phases used in HPLC in terms of resolution (Rs), sensitivity (LOD), efficiency (N), precision (RSD), asymmetry (T) was also performed. This study provided an alternative way for the CEC separation of basic compounds and demonstrated the improvement in the chromatographic parameters.
An amine capillary column was developed in a fused-silica capillary as stationary phase and applied to separation of basic compounds by capillary electrochromatography (CEC). The functionalized capillary was prepared by inmobilization of 3-Aminopropyl)triethoxysilane (APTES). The CEC conditions including APTES and buffer concentration, pH and applied voltage were investigated to obtain the optimal CEC system for the separation of three anesthetics as basic test molecules. The capillary column provided an efficiency of up to 20,000 plates/m. Lidocaine, ketamine and xilacine were baseline separated under the running conditions with 10 mM Na2HPO4 pH 9.0 as BGE. The applied voltage was 5 kV temperature was set at 25oC and UV detection was performed. The resolutions were 4.97 and 1.53 for ketamine, lidocaine and xilacine, respectively. The column used in CEC mode showed better separation of the anesthetics compared with those used in the capillary zone mode. The comparison with reversed stationary phases used in HPLC in terms of resolution (Rs), sensitivity (LOD), efficiency (N), precision (RSD), asymmetry (T) was also performed. This study provided an alternative way for the CEC separation of basic compounds and demonstrated the improvement in the chromatographic parameters.
Cite this paper
Flor, S. , Huala, J. , Tripodi, V. , Lucangioli, S. (2015) Development of Amine Capillary Column Applied to the Analysis of Basic Compounds by Electrochromatography. Journal of Analytical Sciences, Methods and Instrumentation, 5, 66-73. doi: 10.4236/jasmi.2015.54008.
Flor, S. , Huala, J. , Tripodi, V. , Lucangioli, S. (2015) Development of Amine Capillary Column Applied to the Analysis of Basic Compounds by Electrochromatography. Journal of Analytical Sciences, Methods and Instrumentation, 5, 66-73. doi: 10.4236/jasmi.2015.54008.
References
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[1] Kirkland, J.J. (2004) Development of Some Stationary Phases for Reversed-Phase High-Performance Liquid Chroma- tography. Journal of Chromatography A, 1060, 9-21.
http://dx.doi.org/10.1016/S0021-9673(04)01892-8 [2] McCalley, D.V. (2010) The Challenges of the Analysis of Basic Compounds by High Performance Liquid Chromato- graphy: Some Possible Approaches for Improved Separations. Journal of Chromatography A, 1217, 858-880. http://dx.doi.org/10.1016/j.chroma.2009.11.068 [3] Ruiz-Angel, M.J., Carda-Broch, S. and Berthod, A. (2006) Ionic Liquids versus Triethylamine as Mobile Phase Addi- tives in the Analysis of β-Blockers. Journal of Chromatography A, 1119, 202-208.
http://dx.doi.org/10.1016/j.chroma.2005.11.132 [4] Al-Hussin, A., Boysen, R.I., Saito, K. and Hearn, M.T.W. (2014) Preparation and Electrochromatographic Charac- terization of New Chiral β-Cyclodextrin Poly(acrylamidopropyl) Porous Layer Open Tubular Capillary Columns. Jour- nal of Chromatography A, 1358, 199-207. [5] Olsson, J. and Blomberg, L.G. (2008) Enantioseparation of Omeprazole and Its Metabolite 5-Hydroxyomeprazole Using Open Tubular Capillary Electrochromatography with Immobilized Avidin as Chiral Selector. Journal of Chro- matography B: Analytical Technologies in the Biomedical and Life Sciences, 875, 329-332. http://dx.doi.org/10.1016/j.jchromb.2008.05.050 [6] The United States Pharmacopeia, T.-F.R. (2008) Lidocaine. 2527-2528. [7] Yuan, R., Wang, Y. and Ding, G. (2010) Enantiomeric Separation by Capillary Electrochromatography on a Sulfated Poly β-Cyclodextrin Modified Silica-Based Monolith. Analytical Sciences: The International Journal of the Japan Society for Analytical Chemistry, 26, 943-947.
http://dx.doi.org/10.2116/analsci.26.943 [8] The United States Pharmacopeia, T.-F.R. (2008) Physical Test <621>. 232-243. [9] Rathore, A.S. and Horváth, C. (1996) Separation Parameters via Virtual Migration Distances in High-Performance Liquid Chromatography, Capillary Zone Electrophoresis and Electrokinetic Chromatography. Journal of Chromatography A, 743, 231-246. http://dx.doi.org/10.1016/0021-9673(96)00306-8