Insights into head-column field-amplified sample stacking: Part I. Detailed study of electrokinetic injection of a weak base across a short water plug.

Sesták, Jozef; Thormann, Wolfgang (2017). Insights into head-column field-amplified sample stacking: Part I. Detailed study of electrokinetic injection of a weak base across a short water plug. Journal of chromatography A, 1502, pp. 51-61. Elsevier Science 10.1016/j.chroma.2017.04.041

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The fundamentals of electrokinetic injection of the weak base methadone across a short water plug into a phosphate buffer at low pH were studied experimentally and with computer simulation. The current during electrokinetic injection, the formation of the analyte zone, changes occurring within and around the water plug and mass transport of all compounds in the electric field were investigated. The impact of water plug length, plug injection velocity, and composition of sample, plug and background electrolyte are discussed. Experimental data revealed that properties of sample, water plug and stacking boundary are significantly and rapidly altered during electrokinetic injection. Simulation provided insight into these changes, including the nature of the migrating boundaries and the stacking of methadone at the interface to a newly formed phosphoric acid zone. The data confirm the role of the water plug to prevent contamination of the sample by components of the background electrolyte and suggest that mixing caused by electrohydrodynamic instabilities increases the water plug conductivity. The sample conductivity must be controlled by addition of an acid to prevent generation of reversed flow which removes the water plug and to create a buffering environment. Results revealed that a large increase in background electrolyte concentration is not accompanied with a significant increase in stacking.

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