title: A simple LC-MS/MS method for the determination of cortisol, cortisone and tetrahydro-metabolites in human urine: assay development, validation and application in depression patients.
abstract: Chronic stress as well as major depressive disorders is associated with cortisol metabolism. Two enzymes modulate cortisol (F) and cortisone (E) interconversion: 11β-hydroxysteroid dehydrogenase type 1 and type 2 (11β-HSD1 and 11β-HSD2). Furthermore, F and E were inactivated by 5α and 5β reductases to their tetrahydro-metabolites: tetrahydrocortisol (THF), allo-tetrahydrocortisol (5α-THF) and tetrahydrocortisone (THE). To better understand depression a LC-MS/MS method for simultaneous determination of F, E THF, 5α-THF and THE in human urine has been developed and validated. The quantification range was 0.1-160 ng mL(-1) for F and E, and 0.2-160 ng mL(-1) for the tetrahydro-metabolites, with >86.1% recovery for all analytes. The nocturnal urine concentrations of F, E and tetrahydro-metabolites in 12 apparently healthy male adult volunteers and 12 drug-free male patients (age range, 20-50 years) with a diagnosis of depression were analyzed. A series of significant changes in glucocorticoid metabolism can be detected: F/E ratios and (THF+5α-THF)/THE ratios as well as F and THF concentrations were significantly higher in depression patients than in healthy subjects (p<0.05); 5α-THF/F ratios, 5α-THF/THF ratios as well as 5α-THF concentrations were significantly lower in depression patients (p<0.05). The results pointed to the decreased 11β-HSD2 activity and a dysfunction in the 5α-reductase pathway in depressed patients. This method allows the assessment of 11β-HSD1/2 and 5α/β-reductase activities in a single analytical run providing an innovative tool to explain the potential etiology of depression.
authors: Department of Pharmacy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, People's Republic of China.
authors: Xuejia
authors: X
authors: Zhai
authors: Department of Pharmacy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, People's Republic of China.
authors: Fen
authors: F
authors: Chen
authors: Department of Pharmacy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, People's Republic of China.
authors: Chaoran
authors: C
authors: Zhu
authors: Department of Pharmacy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, People's Republic of China. Electronic address: luyn_union@163.com.
authors: Yongning
authors: Y
authors: Lu
mesh_terms: D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1 / Q000378:metabolism; D043209:11-beta-Hydroxysteroid Dehydrogenase Type 2 / Q000378:metabolism; D000328:Adult; D016022:Case-Control Studies; D042944:Cholestenone 5 alpha-Reductase / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods; D003348:Cortisone / Q000737:chemistry* / Q000378:metabolism / Q000652:urine*; D003865:Major Depressive Disorder / Q000378:metabolism / Q000652:urine*; D005938:Glucocorticoids / Q000737:chemistry / Q000378:metabolism / Q000652:urine; D006801:Humans; D006854:Hydrocortisone / Q000737:chemistry* / Q000378:metabolism / Q000652:urine*; D008297:Male; D008875:Middle Aged; D053719:Tandem Mass Spectrometry / Q000379:methods; D013760:Tetrahydrocortisol / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism / Q000652:urine; D013761:Tetrahydrocortisone / Q000737:chemistry* / Q000378:metabolism / Q000652:urine*; D055815:Young Adult
chemical_list: D005938:Glucocorticoids; C006461:allotetrahydrocortisol; D013761:Tetrahydrocortisone; D013760:Tetrahydrocortisol; D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1; D043209:11-beta-Hydroxysteroid Dehydrogenase Type 2; C468387:HSD11B2 protein, human; D042944:Cholestenone 5 alpha-Reductase; D003348:Cortisone; D006854:Hydrocortisone
keywords: 11β-HSD; Cortisol; Depression; LC–MS/MS; Reductase