Experimental design optimization of the capillary electrophoresis separation of leucine enkephalin and its immune complex

Sheikh Md Enayetul Babar, Eun Joo Song, Md Nabiul Hasan, Young Sook Yoo

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

To optimize the capillary electrophoretic separation conditions for leucine enkephalin (LE) and the immune complex of the LE and anti-LE reaction, an analysis using a three-level, three-factorial Box-Behnken design was performed. Three separation parameters, buffer pH (X1), buffer concentration (X2), and applied voltage (X3), were chosen to observe the effect on separation responses. The responses were theoretical plate number, migration time of the LE peak, and resolution between the peaks. The optimum conditions and process validation were determined using statistical regression analysis and surface plot diagrams. The capillary electrophoresis optimum separation conditions were established to be 75 mM phosphate buffer at pH 7.00 with an applied separation voltage of 15 kV. By using the analysis technique, the prediction of responses was satisfactory and process verification yielded values within the ±5% range of the predicted efficiency.

Original languageEnglish
Pages (from-to)2311-2319
Number of pages9
JournalJournal of Separation Science
Volume30
Issue number14
DOIs
StatePublished - Sep 2007

Keywords

  • Box - Behnken design
  • Capillary electrophoresis
  • Leucine enkephalin

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