Identification of endogenous and exogenous glucocorticoids by HPLC-MS in human urine

• Published in: Pharmaceutical chemistry journal Vol. 44; no. 6; pp. 328 - 333
• Main Authors: Dikunets, M. A, Appolonova, S. A, Rodchenkov, G. M
• Format: Journal Article
• Language: English
• Published: Boston Boston : Springer US 01-10-2010; Springer US; Springer
• Subjects: adrenal cortex hormones; Chromatography; Detectors; Dexamethasone; doping control; Formic acid; HPLC-MS; Ionization; Mass spectrometry; Medicine; Methods of Analysis and Process Control; Nitriles; Organic acids; Organic Chemistry; Pharmacology/Toxicology; Pharmacy; Structure of Chemical Compounds; Toluene; corticosteroids; HPLC-MS; doping control
• ISSN: 0091-150X; 1573-9031
• DOI: 10.1007/s11094-010-0461-2

A selective and sensitive screening method for identifying 20 endogenous and exogenous corticosteroids in human urine is presented. Analysis was performed by HPLC with a mass-selective “ion trap” detector and spray ionization. Urine samples were prepared by liquid-liquid extraction (with a mixture of toluene and di-ethyl ether, 50:50 v/v). The dry residue was redissolved in 50 μl of methanol and aliquots of 5 μl were loaded onto the HPLC-MS system. Chromatographic separation was performed on a Restek Ultra C18 (2.1 × 100 mm, 5 μm) column. The mobile phase was a solution of 0.05% formic acid containing 20 mM ammonium acetate pH 3 (A) and acetonitrile (B). The detection limits for all corticosteroids were 1 - 3 ng/ml. Chromatographic separation and mass spectrometric data for identification of mixtures of steric isomers of betamethasone and dexamethasone are presented.