The effect of N,N,N',N'-tetramethyl-1,3-butanediamine (TMBD) in the running electrolyte on the electroosmotic flow and the migration behavior of four standard basic proteins in bare fused-silica capillaries was examined at pH 4.0, 5.5, and 6.5. Depending on the electrolyte pH and additive concentration the electroosmotic flow was either cathodic or anodic. A similar Langmuirian-type dependence of the electroosmotic flow on the concentration of TMBD in the running electrolyte was found at the three experimented pH values, which may be indicative of the specific adsorption of the additive in the immobilized region of the electric double layer at the interface between the capillary wall and the electrolyte solution. Electrophoretic separations of the four standard basic proteins performed at the three above pH values, showed well-resolved, efficient and symmetric peaks, demonstrating the utility of this additive for protein electrophoresis in bare fused-silica capillaries. The variations in separation efficiency, peak capacity, resolution and reproducibility of migration times as a function of the additive concentration at pH 6.5 were also examined.
The electrophoresis of epsilon-N-2-furoylmethyl-L-lysine (furosine) was studied in an attempt to develop a method for the identification and quantitation of this compound in processed food. The effect of pH and composition of electrolyte solution on both the electrophoretic migration of furosine and the electroosmotic flow in a bare fused-silica capillary of 75 mu m internal diameter was investigated. We demonstrate that the addition, to the running electrolyte solution, of N,N,N',N'-tetramethyl-1,3-butanediamine (TMBD) at concentrations ranging from 20 to 80 mM improves peak efficiency and can be used to modulate the migration time of furosine by controlling the electroosmotic flow which is reversed from cathodic to anodic. In a sample of dried milk subjected to a long period of storage under controlled conditions, furosine could be efficiently and reproducibly separated and quantitated by employing as the running electrolyte 60 mM TMBD titrated to pH 2.5. Capillary electrophoresis is a promising technique for the rapid identification and quantitation of furosine in processed food.
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