Determination of dissociation constants of cytokinins by capillary zone electrophoresis.
A method for the pKa determination, based on mobility data measured by capillary zone electrophoresis, was applied to cytokinins and their analogs. The combination of charged mobility standards with an uncharged electroosmosis marker, injected in the uncoated capillary simultaneously with the measured substances, allows one to minimize the number of runs, reduce their duration and, in addition, to inform on the run-to-run stability of electroosmosis and on contingent side-effects. pKa values of investigated cytokinins and their analogs ranged from 2.8 to 4.0 at 25 degrees C in the phosphate and acetate buffers of ionic strength 0.015 M. Standard deviations of the constants, obtained by the non-linear fitting of equations for the pKa calculation, were 3-5-times lower than standard deviations from the linear fitting or from the point-to-point calculation utilizing the Hendersson-Haselbalch equation. The equation of Boltzman sigmoid offers two checks on reliability of effective mobilities that serve as the raw data in the pKa calculation.
Barták
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P
Center of Bioanalytical Research, Palacký University, Olomouc, Czech Republic.
Bednár
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P
Stránský
Z
Z
Bocek
P
P
Vespalec
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R
D003583:Cytokinins / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*
D003583:Cytokinins