Sodium glycylglycine as effective electrolyte run buffer for ascorbic and uric acid separation by CZE: a comparison with two other CE assays.

Angelo Zinellu, Salvatore Sotgia, Salvatore Caddeo, Luca Deiana, Ciriaco Carru

Index: J. Sep. Sci. 28 , 2193-2199, (2005)

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Abstract

A new CE method for ascorbic acid (AA) and uric acid (UA) detection in human plasma has been developed. Analytes were resolved in less than 4 min by employing sodium glycylglycine (Glygly) as electrolyte run buffer at pH 8.0. Using the diode array detector ability to measure multiple wavelengths simultaneously, detection was optimized by monitoring the run at 262 nm for AA and at 288 nm for UA. Electrophoretic parameters such as resolution, migration times, efficiency, and peak areas of this new method were compared to those obtained by the two CE assays described in literature, in which the analytes separation was achieved by using sodium borate (that allows faster migration times but poor resolution) or tricine (with the highest resolution but elevated migration times) as electrolyte run buffer. Sodium Glygly allows to obtain the same good resolution given by the tricine buffer but with the faster analysis times of the sodium borate run buffer. Ascorbate and urate levels were measured in 35 healthy volunteers by the three methods and the obtained data were compared by three different statistical tests (mountain plot, Passing-Bablok regression, and Bland-Altman test) in order to verify the accuracy of our proposed method.