Determination of free tetrahydrocortisol and tetrahydrocortisone ratio in urine by liquid chromatography-tandem mass spectrometry

Scandinavian Journal of Clinical and Laboratory Investigation
U TurpeinenE Hämäläinen

Abstract

Measurement of urinary free tetrahydrocortisol and tetrahydrocortisone ratio (allo-THF+THF)/THE is clinically important in the diagnosis of hypertension caused by congenital absence of 11beta-hydroxysteroid dehydrogenase type 2 (apparent mineralocorticoid excess, AME) or inhibition of the enzyme after licorice ingestion. Although gas chromatography-mass spectrometry (GC-MS) provides reliable results, it requires derivatization and is lengthy and time-consuming. The purpose of this study was to demonstrate that detection by liquid chromatography-mass spectrometry (LC-MS) is a potentially superior method. The analysis utilizes 1 mL urine. The samples were extracted with solid-phase extraction (SPE) using ethyl acetate as eluent. The extract was evaporated to dryness, and allo-tetrahydrocortisol (allo-THF), THF and THE concentrations were analyzed by LC-MS/MS operating in the negative mode after separation on a reversed-phase column. The calibration curves exhibited consistent linearity and reproducibility in the range of 7.5-120 nmol/L. Interassay CVs were 7.0-10 % at mean ratios of (allo-THF+THF)/THE of 0.54-1.9. The detection limit of the analytes was 0.4-0.8 nmol/L (signal-to-noise ratio = 3). The mean recovery of the three an...Continue Reading

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Citations

Sep 1, 2009·Journal of Clinical Research in Pediatric Endocrinology·John W Honour
Jun 17, 2016·Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment·Luca ChiesaFrancesco Arioli
Dec 1, 2006·Journal of Mass Spectrometry : JMS
May 1, 2014·Expert Review of Endocrinology & Metabolism·Yuhei KoyamaTomonobu Hasegawa
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Apr 1, 2014·Chromatographia·Fidel AllendeCarlos E Fardella
Jun 12, 2018·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Takaaki MatsumotoTatsuya Higashi
Jun 2, 2018·Pain Research & Management : the Journal of the Canadian Pain Society = Journal De La Société Canadienne Pour Le Traitement De La Douleur·Kordian StaniszewskiAnnika Rosén

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