Study of off-line combinations of HPLC and CZE methods for analysis of low molecular weight substances potentially related to humic substances degradation products
2009
Hutta, M., Comenius Univ., Bratislava (Slovak Republic) | Kaniansky, D., Comenius Univ., Bratislava (Slovak Republic) | Góra, R., Comenius Univ., Bratislava (Slovak Republic)
Design and development of novel systems for characterization of humic substances and/or their degradation products in solutions is still challenging. Two-dimensional system devised in this work was created by off-line combination of gradient elution reversed-phase high performance liquid chromatography and capillary zone electrophoresis in an open system. Electrophoretic system was used as a selective detector for collected liquid chromatographic fractions. We tested orthogonality of liquid chromatography and zone electrophoresis for group of cationogenic and anionogenic compounds under conditions of high compatibility of both separation systems. The choice of the model substances was based on criteria of their potential presence in solutions of degradation products of humic substances (photolysis, hydrolysis, oxidation, oxidative photolysis, biodegradation etc.). Migration and retention characteristics of analytes were evaluated both by Spearman rank correlation coefficients and Pearsons correlation coefficients. We measured migration times in electrophoresis and retention times in chromatography, respectively, of basic compounds by use of phosphate buffers of varying pH values. Similar experiments were done also with the set of acidic compounds. Combination of reversed - phase, or aqueous normal phase liquid chromatography, respectively, and capillary electrophoresis gives us results, that are interpreted as non-existence of proof of data sets correlations in studied combinations of CZE and HPLC systems. That practically means formulation of separation systems orthogonality. Similar combinations of orthogonal separation systems will be used also for the characterization of treated solutions of humic substances.
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