title: An LC-MS/MS method for steroid profiling during adrenal venous sampling for investigation of primary aldosteronism.
abstract: BACKGROUND
  Steroid profiling for diagnosis of endocrine disorders featuring disordered production of steroid hormones is now possible from advances in liquid chromatography with tandem mass spectrometry (LC-MS/MS). Adrenal venous (AV) measurements of aldosterone and cortisol are a standard practice in the clinical work-up of primary aldosteronism, but do not yet take advantage of steroid profiling.
  
  
  METHODS
  A novel LC-MS/MS based method was developed for simultaneous measurement of 15 adrenal steroids: aldosterone, corticosterone, 11-deoxycorticosterone, progesterone, pregnenolone, cortisone, cortisol, 11-deoxycortisol, 17-hydroxyprogesterone, androstenedione, dehydroepiandrosterone, dehydroepiandrosterone-sulfate, 21-deoxycortisol, 18-oxocortisol and 18-hydroxycortisol. These were compared in peripheral venous (pV) and AV plasma from 70 patients undergoing AV sampling with and without cosyntropin stimulation. Aldosterone and cortisol levels measured by LC-MS/MS were compared with those measured by immunoassay.
  
  
  RESULTS
  Reproducibility of measurements with coefficients of variation ≤10% as well as analytical sensitivity sufficient to measure low pV levels particularly of aldosterone demonstrate the utility of the assay for profiling adrenal steroids in primary aldosteronism. Method comparisons indicated assay and concentration dependent differences of cortisol and aldosterone concentrations measured by immunoassay and LC-MS/MS. Median AV/pV ratios of 11-deoxycortisol (53.0), 17-hydroxyprogesterone (33.4), pregnenolone (62.4), androstenedione (40.6) and dehydroepiandrosterone (33.3) were 2.9- to, 5.4-fold larger than those for cortisol (11.6), with additionally generally larger increases than for cortisol with than without cosyntropin stimulation.
  
  
  CONCLUSIONS
  Our LC-MS/MS assay, in addition to improvements over existing immunoassay measurements of aldosterone and cortisol, offers profiling of 13 other adrenal steroids, providing a potentially useful method for the clinical work-up of patients with primary aldosteronism. In particular, the larger AV/pV ratios of several steroids compared to cortisol suggest more sensitive alternatives to the latter for assessing positioning of AV sampling catheters.
authors.affiliation: Institute of Clinical Chemistry and Laboratory Medicine, University Hospital Carl Gustav Carus, Medical Faculty Carl Gustav Carus, Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. Electronic address: Mirko.Peitzsch@uniklinikum-dresden.de.
authors.forename: Mirko
authors.initials: M
authors.lastname: Peitzsch
authors.affiliation: Department of General Internal Medicine, Radboud University Medical Center, Geert Grooteplein 8, 6525 Nijmegen, The Netherlands.
authors.forename: Tanja
authors.initials: T
authors.lastname: Dekkers
authors.affiliation: Heinrich-Heine-University of Dusseldorf, Medical Faculty, Division for Specific Endocrinology, Moorenstrasse 5, 40225 Dusseldorf, Germany; Department of Medicine III, University Hospital Carl Gustav Carus, Medical Faculty Carl Gustav Carus, Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Haase
authors.affiliation: Department of Laboratory Medicine, Radboud University Medical Center, Geert Grooteplein 10, 6525 Nijmegen, The Netherlands.
authors.forename: Fred C G J
authors.initials: FC
authors.lastname: Sweep
authors.affiliation: Heinrich-Heine-University of Dusseldorf, Medical Faculty, Department of Nephrology, Moorenstrasse 5, 40225 Dusseldorf, Germany.
authors.forename: Ivo
authors.initials: I
authors.lastname: Quack
authors.affiliation: Heinrich-Heine-University of Dusseldorf, Medical Faculty, Department of Diagnostic and Interventional Radiology, Moorenstrasse 5, 40225 Dusseldorf, Germany.
authors.forename: Gerald
authors.initials: G
authors.lastname: Antoch
authors.affiliation: Institute of Clinical Chemistry and Laboratory Medicine, University Hospital Carl Gustav Carus, Medical Faculty Carl Gustav Carus, Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Siegert
authors.affiliation: Department of General Internal Medicine, Radboud University Medical Center, Geert Grooteplein 8, 6525 Nijmegen, The Netherlands; Department of Medicine III, University Hospital Carl Gustav Carus, Medical Faculty Carl Gustav Carus, Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany.
authors.forename: Jacques W M
authors.initials: JW
authors.lastname: Lenders
authors.affiliation: Department of General Internal Medicine, Radboud University Medical Center, Geert Grooteplein 8, 6525 Nijmegen, The Netherlands.
authors.forename: Jaap
authors.initials: J
authors.lastname: Deinum
authors.affiliation: Heinrich-Heine-University of Dusseldorf, Medical Faculty, Division for Specific Endocrinology, Moorenstrasse 5, 40225 Dusseldorf, Germany; Department of Medicine III, University Hospital Carl Gustav Carus, Medical Faculty Carl Gustav Carus, Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany.
authors.forename: Holger S
authors.initials: HS
authors.lastname: Willenberg
authors.affiliation: Institute of Clinical Chemistry and Laboratory Medicine, University Hospital Carl Gustav Carus, Medical Faculty Carl Gustav Carus, Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany; Department of Medicine III, University Hospital Carl Gustav Carus, Medical Faculty Carl Gustav Carus, Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany.
authors.forename: Graeme
authors.initials: G
authors.lastname: Eisenhofer
mesh_terms: D000311:Adrenal Glands / Q000098:blood supply / Q000378:metabolism*; D000328:Adult; D000368:Aged; D002853:Chromatography, Liquid / Q000379:methods*; D003366:Cosyntropin / Q000097:blood; D005260:Female; D006801:Humans; D006854:Hydrocortisone / Q000097:blood; D006929:Hyperaldosteronism / Q000175:diagnosis* / Q000378:metabolism; D007118:Immunoassay; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results; D013256:Steroids / Q000032:analysis* / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D013256:Steroids; D003366:Cosyntropin; D006854:Hydrocortisone
keywords: Adrenal venous sampling; LC–MS/MS; Primary aldosteronism; Steroid profiling