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Micellar electrokinetic capillary chromatography (MECC) of UV-absorbing constituents in normal urine: a chemometric optimisation of the separation.
Electrophoresis. 1997 Sep; 18(10):1847-56.E

Abstract

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, beta-cyclodextrin (beta-CD) concentration, voltage, pH, temperature and electrolyte additives (urea, beta-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 microns x 44 cm (37 cm effective length) fused silica capillary the final optimum conditions were found to be, an electrolyte consisting of 30 mM sodium tetraborate, pH 10, containing 75 mM SDS and 10 mM beta-CD, 15 degrees 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.

Authors+Show Affiliations

Alfazema LN
Department of Chemistry, University of Greenwich, London, UK.
Hows ME
No affiliation info available
Howells S
No affiliation info available
Perrett D
No affiliation info available

MeSH

Analysis of VarianceBuffersChromatographyChromatography, High Pressure LiquidElectric ConductivityElectrolytesElectrophoresis, CapillaryFemaleHumansHydrogen-Ion ConcentrationMaleReproducibility of ResultsTime FactorsUltraviolet RaysUrine

Pub Type(s)

Journal Article

Language

eng

PubMed ID

9372279

Citation

Alfazema, L N., et al. "Micellar Electrokinetic Capillary Chromatography (MECC) of UV-absorbing Constituents in Normal Urine: a Chemometric Optimisation of the Separation." Electrophoresis, vol. 18, no. 10, 1997, pp. 1847-56.
Alfazema LN, Hows ME, Howells S, et al. Micellar electrokinetic capillary chromatography (MECC) of UV-absorbing constituents in normal urine: a chemometric optimisation of the separation. Electrophoresis. 1997;18(10):1847-56.
Alfazema, L. N., Hows, M. E., Howells, S., & Perrett, D. (1997). Micellar electrokinetic capillary chromatography (MECC) of UV-absorbing constituents in normal urine: a chemometric optimisation of the separation. Electrophoresis, 18(10), 1847-56.
Alfazema LN, et al. Micellar Electrokinetic Capillary Chromatography (MECC) of UV-absorbing Constituents in Normal Urine: a Chemometric Optimisation of the Separation. Electrophoresis. 1997;18(10):1847-56. PubMed PMID: 9372279.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Micellar electrokinetic capillary chromatography (MECC) of UV-absorbing constituents in normal urine: a chemometric optimisation of the separation. AU - Alfazema,L N, AU - Hows,M E, AU - Howells,S, AU - Perrett,D, PY - 1998/2/12/pubmed PY - 1998/2/12/medline PY - 1998/2/12/entrez SP - 1847 EP - 56 JF - Electrophoresis JO - Electrophoresis VL - 18 IS - 10 N2 - 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, beta-cyclodextrin (beta-CD) concentration, voltage, pH, temperature and electrolyte additives (urea, beta-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 microns x 44 cm (37 cm effective length) fused silica capillary the final optimum conditions were found to be, an electrolyte consisting of 30 mM sodium tetraborate, pH 10, containing 75 mM SDS and 10 mM beta-CD, 15 degrees 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. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/9372279/Micellar_electrokinetic_capillary_chromatography__MECC__of_UV_absorbing_constituents_in_normal_urine:_a_chemometric_optimisation_of_the_separation_ L2 - https://doi.org/10.1002/elps.1150181021 DB - PRIME DP - Unbound Medicine ER -