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Investigation of the factors that induce analyte peak splitting in capillary electrophoresis
Authors:Chen Xiaofeng  Xie Jianping  Li Cunhong  Hu Zhide  Chen Xingguo
Affiliation:Department of Chemistry, Lanzhou University, Lanzhou, 730000, P. R. China. chenxf@lzu.edu.cn
Abstract:Peak splitting has a detrimental effect on analyses by capillary electrophoresis. Many papers have reported it and several mechanisms have been proposed to explain the phenomenon. We investigated the electrophoretic behavior of an amphoteric analyte, levodopa, in phosphate buffer and observed a peak splitting phenomenon at moderate sample concentrations and under general analytical conditions, even without organic solvent. The dependence of effective mobility on pH was taken into account and pKa values of 2.30, 8.11, and 9.92 were obtained for levodopa. Then, we constructed pH-dependent distribution diagrams of levodopa and phosphate species present in aqueous solution and proposed that the most relevant factors contributing to peak splitting are the presence of ionizable groups in the analyte molecule and the occurrence of ionization, yielding charged species which interacted with buffer electrolyte species in a definite pH range to form complexes. This result is different from those presented in the literature and broadens our understanding of amphoteric analyte peak splitting.
Keywords:Capillary electrophoresis  Peak splitting  Ampholyte  Levodopa  Apparent dissociation constant  Isoelectric point
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