Wj. Jusko et al., 15 YEARS OF OPERATION OF A HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ASSAY FOR PREDNISOLONE, CORTISOL AND PREDNISONE IN PLASMA, Journal of chromatography B. Biomedical applications, 658(1), 1994, pp. 47-54
Citations number
29
Categorie Soggetti
Chemistry Analytical
Journal title
Journal of chromatography B. Biomedical applications
A high-performance liquid chromatographic (HPLC) assay first described
in 1979 has been modified and revalidated for the simultaneous determ
ination of prednisone, cortisol and prednisolone in human plasma using
betamethasone as an internal standard. Revisions include: mobile phas
e composition; use of a precolumn, automated injector, integrator, and
computer software; improved sensitivity and quantitation; thorough in
vestigation of stability, variation, and specimen type; and inclusion
of suggested quality control criteria. Plasma-based drug standards are
extracted with methylene chloride and washed with sodium hydroxide fo
llowed by a water wash. After evaporation of solvent and reconstitutio
n with mobile phase, the extracts are then injected onto a silica gel
column (Zorbax SIL) for chromatography with UV absorbance at 254 nm. C
alculated limits of quantitation are 10 ng/ml and limits of detection
are less than 5 ng/ml. Intra- and inter-day coefficients of variation
for quality control samples for all three corticosteroids are less tha
n 11.2%. Recovery and stability data are also provided. Several drugs
that may be coadministered do not interfere with the analysis.