title: Simultaneous determination of urinary cortisol, cortisone and corticosterone in parachutists, depressed patients and healthy controls in view of biomedical and pharmacokinetic studies.
abstract: A rapid and sensitive reversed-phase liquid chromatographic method (RP-LC) with UV detection has been developed for the determination of free cortisol, cortisone and corticosterone in human urine. The assay was performed after a solid-phase extraction procedure (SPE) with dexamethasone as the internal standard. Chromatographic separation was carried out on a Nucleosil 100 C(18) analytical column using a mixture of acetonitrile and water (30 : 70, v/v) as a mobile phase at a flow-rate of 1 mL min(-1). Spectrophotometric detection was performed at 240 nm. The method has been validated for accuracy, precision, selectivity, linearity, recovery and stability. The absolute recoveries of glucocorticoids were above 94.6%. The limits of detection (LOD) and quantification (LOQ) were 0.5 and 2 ng mL(-1), respectively, for all analytes. Linearity was confirmed in the range of 2-300 ng mL(-1) with a correlation coefficient greater than 0.9997 for all steroid hormones. The proposed method was sensitive, robust and specific allowing reliable quantification of steroid hormones. This method was successfully applied for determination of three endogenous glucocorticoid levels in human urine. The studies were performed on 20 sedentary healthy volunteers in comparison to two socially diversified groups, namely 10 parachutists before and after jump and 10 patients with depression. Pharmacokinetic studies performed on these groups indicated that urinary free cortisol and cortisol-to-cortisone ratios can be treated as biomarkers of stress and depressive disorders.
authors.affiliation: Department of Pharmaceutical Chemistry, Medical University of Gdańsk, Hallera 107, 80-416 Gdańsk, Poland.
authors.forename: Alina
authors.initials: A
authors.lastname: Plenis
authors.forename: Lucyna
authors.initials: L
authors.lastname: Konieczna
authors.forename: Ilona
authors.initials: I
authors.lastname: Olędzka
authors.forename: Piotr
authors.initials: P
authors.lastname: Kowalski
authors.forename: Tomasz
authors.initials: T
authors.lastname: Bączek
mesh_terms: D000704:Analysis of Variance; D001359:Aviation; D056148:Chromatography, Reverse-Phase; D003345:Corticosterone / Q000737:chemistry / Q000652:urine*; D003348:Cortisone / Q000737:chemistry / Q000652:urine*; D003866:Depressive Disorder / Q000652:urine*; D003907:Dexamethasone / Q000493:pharmacokinetics / Q000652:urine; D005938:Glucocorticoids / Q000493:pharmacokinetics / Q000652:urine; D006801:Humans; D006854:Hydrocortisone / Q000737:chemistry / Q000652:urine*; D015394:Molecular Structure; D025341:Principal Component Analysis; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D013056:Spectrophotometry, Ultraviolet
chemical_list: D005938:Glucocorticoids; D003907:Dexamethasone; D003348:Cortisone; D003345:Corticosterone; D006854:Hydrocortisone