Characterization of buffers for electrokinetic separations |
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Authors: | Subir K Basak Ajoy Velayudhan Michael R Ladisch |
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Institution: | (1) Department of Agricultural Engineering, Purdue University, 1295 Potter Center, 47907-1295 West Lafayette, IN;(2) Laboratory of Renewable Resources Engineering, Purdue University, 1295 Potter Center, 47907-1295 West Lafayette, IN;(3) Department of Agricultural Engineering, Purdue University, 1295 Potter Center, 47907-1295 West Lafayette, IN |
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Abstract: | Buffers used in electrophoresis and electrochromatography must have a relatively low ionic strength in order to minimize ohmic
heating in the presence of an applied potential. Calculation of pH, ionic strength, and the van Slyke buffer capacity, β,
is therefore important. This paper describes thea priori calculation of these parameters for tris buffer made up with either glycine (a zwitterion) or HCl. A quadratic expression
for pH, valid over wide ranges, is obtained for both buffer systems. The calculated values of pH, ionic strength, and buffer
capacity are shown to agree with experimental results as a function of tris, HCl, and glycine concentrations ranging from
1 to 50 mM. A new parameter, the electrokinetic buffer effectiveness factor, is introduced to characterize buffers being considered
for use in electrokinetic systems such as electrochromatography, and is used to determine the appropriate composition ranges
for the buffer components. |
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Keywords: | Zwitterions buffer electrochromatography pH buffer-capacity ionic strength tris glycine HCL |
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