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.

The comparison of two column classification systems during the chromatographic analysis of steroids

“…In the first one, a new SPE‐LC‐UV method with an NUC18‐125/5 column was developed and validated in accordance to FDA and ICH guidelines. In this method, similar sample preparation procedure to that one described in 39 was applied but slight modification was introduced (dexamethasone as I.S. was added to the sample at the level of 400 ng/mL).…”

Antidepressant-like activity and cardioprotective effects of fatty acid amide hydrolase inhibitor URB694 in socially stressed Wistar Kyoto rats

“…After centrifugation (14000 g, 10 min, 4°C), the supernatant was directly injected in the liquid chromatography/tandem mass spectrometry system (HPLC/MS/MS) for quantification of corticosterone levels, in accordance with previously published analytical methods (e.g., Plenis et al, 2011).…”

The 24-hour urinary cortisol in post-traumatic stress disorder: A meta-analysis

“…When 24-h urinary cortisol collection method [40], and storage methods of 24-h urinary cortisol has be unified [41]. The 24-h urinary cortisol could be a quick biomarker assay to assist in screening a possibility for screening a lot of people with PTSD [10,42].…”