Liquid chromatographic analysis of prednisolone, prednisone and their 20-reduced metabolites in perfusion media

• Published in: Journal of chromatography Vol. 563; no. 2; pp. 341 - 347
• Main Authors: Cannell, G.R., Mortimer, R.H., Maguire, D.J., Addison, R.S.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.02.1991; Elsevier
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chromatography, Liquid - instrumentation; Chromatography, Liquid - methods; Female; Fundamental and applied biological sciences. Psychology; Humans; Hydrocortisone - analysis; Hydrocortisone - metabolism; Other biological molecules; Perfusion; Placenta - chemistry; Placenta - metabolism; Prednisolone - analysis; Prednisolone - metabolism; Prednisone - analysis; Prednisone - metabolism; Pregnancy; Terpenes, steroids. Hormones; Reversed phase chromatography; Steroid hormone; Liquid chromatography; Prednisone; Glucocorticoid; Prednisolone; Quantitative analysis
• ISSN: 0378-4347
• DOI: 10.1016/0378-4347(91)80040-J

A reversed-phase liquid chromatographic assay was developed to quantitate prednisolone, prednisone and the 20α-dihydro and 20β-dihydro reduced metabolites of both parent compounds in tissue culture media from in vitro perfusions of the human placental lobule. Steroids were extracted from perfusate, using reversed-phase cartridges, with average recoveries of 95.2% or greater. The internal standard for the analyses was 6α-methylprednisolone. In this assay cortisol coelutes with prednisolone, however, no other significant interferences were found. Assay of each steroid was linear in the range 0–1 μg/ml. Intra-assay coefficients of variation were measured at 10 and 750 ng/ml with ranges of 3.4% (20α-dihydroprednisone) to 8.8% (20β-dihydroprednisolone) and 4.1% (20β-dihydroprednisone) to 8.8% (prednisone). The corresponding inter-assay coefficients of variation were 3.3% (20α-dihydroprednisone) to 9.1% (20β-dihydroprednisolone) and 1.9% (prednisolone) to 3.5% (prednisone). The analyses utilized two C 18 columns which were linked together and maintained at 40°C.