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Micellar electrokinetic capillary chromatography (MECC) of UV‐absorbing constituents in normal urine: A chemometric optimisation of the separation
Author(s) -
Alfazema Luwiza N.,
Hows Mark E. P.,
Howells Sian,
Perrett David
Publication year - 1997
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150181021
Subject(s) - chromatography , micellar electrokinetic chromatography , capillary electrophoresis , chemistry , resolution (logic) , electrolyte , factorial experiment , electrokinetic phenomena , capillary action , analytical chemistry (journal) , materials science , statistics , mathematics , electrode , artificial intelligence , computer science , composite material
A micellar electrokinetic capillary chromatography (MECC) method when compared to free solution capillary electrophoresis (CZE) was shown to offer improved selectivity and resolution for the separation of UV‐absorbing components of human urine. Some of the factors affecting MECC separation e.g. methanol concentration, sodium dodecyl sulphate (SDS) concentration, β‐cyclodextrin (β‐CD) concentration, voltage, pH, temperature and electrolyte additives (urea, β‐CD and Brij 35) were optimised using chemometric techniques. Three‐level three‐factor (3 3 ) factorial designs and simplex optimisation were used to achieve optimised conditions with the goal of obtaining the maximum number of peaks in the shortest possible analysis time. Using a TSP CE2000 instrument with detection from 195–300 nm and fitted with a 75 μm × 44 cm (37 cm effective length) fused silica capillary the final optimum conditions were found to be, an electrolyte consisting of 30 m M sodium tetraborate, pH 10, containing 75 m M SDS and 10 m M β‐CD, 15°C, 20 kV, 4 s hydrodynamic injection of filtered urine. These conditions were capable of separating 70 peaks from a normal human urine pool in less than 12 min. The separation of components in urine using the optimised MECC was simpler, more reproducible, faster and gave better resolution than gradient reversed‐phase high performance liquid chromatography This work was first presented at the 4th International Symposium on Capillary Electrophoresis, University of York, UK, August 1996 .