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Experimental design optimization of the capillary electrophoresis separation of leucine enkephalin and its immune complex
Author(s) -
Babar Sheikh Md. Enayetul,
Song Eun Joo,
Hasan Md. Nabiul,
Yoo Young Sook
Publication year - 2007
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200700145
Subject(s) - capillary electrophoresis , chromatography , factorial experiment , analytical chemistry (journal) , chemistry , response surface methodology , electrophoresis , central composite design , resolution (logic) , capillary action , separation process , linear regression , fractional factorial design , materials science , mathematics , statistics , artificial intelligence , computer science , composite material
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 ( X 1 ), buffer concentration ( X 2 ), and applied voltage ( X 3 ), 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.