Capillary electrochromatography of steroids: Increased sensitivity by on-line concentration and comparison with high-performance liquid chromatography

A reversed-phase HPLC method previously developed for the analysis of progesterone and its major metabolites has been transferred successfully to a capillary electrochromatography (CEC) system. Procedures for fabricating packed capillaries and the modifications made to the capillary electropherograp...

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Bibliographic Details
Published in:Journal of Chromatography A 1998-03, Vol.798 (1), p.259-267
Main Authors: Stead, D.A, Reid, R.G, Taylor, R.B
Format: Article
Language:English
Subjects:
17-alpha-Hydroxyprogesterone - blood
20-alpha-Dihydroprogesterone - blood
Acetonitriles
Analysis
Analytical chemistry
Androstenedione - blood
Biological and medical sciences
Chemistry
Chromatographic methods and physical methods associated with chromatography
Chromatography - methods
Chromatography, High Pressure Liquid
Exact sciences and technology
General pharmacology
Hydrogen-Ion Concentration
Medical sciences
Methanol
Norethindrone - blood
Other chromatographic methods
Pharmacology. Drug treatments
Progesterone
Progesterone - blood
Sensitivity and Specificity
Solvents
Steroids
Steroids - blood
Testosterone - blood
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Summary:A reversed-phase HPLC method previously developed for the analysis of progesterone and its major metabolites has been transferred successfully to a capillary electrochromatography (CEC) system. Procedures for fabricating packed capillaries and the modifications made to the capillary electropherograph which allow operation in the CEC mode without pressurisation are described. The dependence of electroosmotic flow on electric field strength, pH and organic modifier content is discussed. Direct comparison with HPLC shows that CEC provides useful gains in efficiency and speed of analysis and requires vastly reduced amounts of both chromatographic phases and material for analysis. On-line concentration is described which allows the lower sensitivity of CEC to be offset by injecting analytes from a non-eluting solution. Examination of steroids in plasma demonstrates that the superior separation by CEC is maintained in a complex biological matrix.
ISSN:0021-9673
DOI:10.1016/S0021-9673(97)00840-6