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 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.
Stead
D A
DA
School of Pharmacy, Robert Gordon University, Schoolhill, Aberdeen, UK.
Reid
R G
RG
Taylor
R B
RB
D019326:17-alpha-Hydroxyprogesterone / Q000097:blood; D004092:20-alpha-Dihydroprogesterone / Q000097:blood; D000097:Acetonitriles; D000735:Androstenedione / Q000097:blood; D002845:Chromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D006863:Hydrogen-Ion Concentration; D000432:Methanol; D009640:Norethindrone / Q000097:blood; D011374:Progesterone / Q000097:blood; D012680:Sensitivity and Specificity; D012997:Solvents; D013256:Steroids / Q000097:blood*; D013739:Testosterone / Q000097:blood
D000097:Acetonitriles; D012997:Solvents; D013256:Steroids; D004092:20-alpha-Dihydroprogesterone; D013739:Testosterone; D000735:Androstenedione; D011374:Progesterone; D019326:17-alpha-Hydroxyprogesterone; D009640:Norethindrone; D000432:Methanol; C032159:acetonitrile