title: Simultaneous measurement of testosterone, androstenedione and dehydroepiandrosterone (DHEA) in serum and plasma using Isotope-Dilution 2-Dimension Ultra High Performance Liquid-Chromatography Tandem Mass Spectrometry (ID-LC-MS/MS).
abstract: The adrenal and gonadal androgens, testosterone, androstenedione and dehydroepiandrosterone (DHEA) play an important role in sexual development as well as in other processes. We developed a method for simultaneous quantitative analysis of serum and plasma testosterone, androstenedione and DHEA levels using Isotope-Dilution Liquid-Chromatography Tandem Mass Spectrometry (ID-LC-MS/MS). Samples underwent liquid-liquid extraction and were analyzed on an Acquity 2D-UPLC-System and a Xevo TQ-S tandem mass spectrometer (Waters). The intra-assay and inter-assay coefficients of variation were <4.0%, <6.3% and <7.0% and <6.0%, <8.1% and <7.7% for testosterone, androstenedione and DHEA, respectively. Inter-assay CVs at the lower limit were 10.6%, 16.9% and 9.0% for testosterone (0.10nmol/L), androstenedione (0.10nmol/L) and DHEA (1.0nmol/L), respectively. Recoveries of spiked analytes were 93-107%. The present testosterone method compared well (y=1.00x-0.04; r=0.998) to a published ID-LC-MS/MS method for testosterone in our lab. The latter method being concordant with a published reference method (Bui et al., 2013). The present method compared well to a published ID-LC-MS/MS method (Kushnir et al., 2010) (y=1.06x-0.06; r=0.996 for testosterone; y=1.04x-0.04; r=0.995 for androstenedione and y=1.03x+0.01; r=0.991 for DHEA). In conclusion, we developed a sensitive and accurate ID-LC-MS/MS method to simultaneously measure serum testosterone, androstenedione and DHEA in serum and plasma.
authors.affiliation: Department of Clinical Chemistry, Endocrine Laboratory, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Rahel M
authors.initials: RM
authors.lastname: Büttler
authors.affiliation: Department of Clinical Chemistry, Endocrine Laboratory, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Frans
authors.initials: F
authors.lastname: Martens
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology and Department of Pathology, University of UT, Salt Lake City, USA.
authors.forename: Mark M
authors.initials: MM
authors.lastname: Kushnir
authors.affiliation: Department of Clinical Chemistry, Laboratory of Endocrinology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Mariette T
authors.initials: MT
authors.lastname: Ackermans
authors.affiliation: Department of Clinical Chemistry, Endocrine Laboratory, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Marinus A
authors.initials: MA
authors.lastname: Blankenstein
authors.affiliation: Department of Clinical Chemistry, Endocrine Laboratory, VU University Medical Center, Amsterdam, The Netherlands. Electronic address: a.heijboer@vumc.nl.
authors.forename: Annemieke C
authors.initials: AC
authors.lastname: Heijboer
mesh_terms: D000735:Androstenedione / Q000097:blood*; D002853:Chromatography, Liquid / Q000379:methods*; D003687:Dehydroepiandrosterone / Q000097:blood*; D006801:Humans; D007201:Indicator Dilution Techniques; D059625:Liquid-Liquid Extraction; D010949:Plasma / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013739:Testosterone / Q000097:blood*
chemical_list: D013739:Testosterone; D000735:Androstenedione; D003687:Dehydroepiandrosterone
keywords: Androgens; Androstenedione; Dehydroepiandrosterone (DHEA); Liquid chromatography; Mass spectrometry; Testosterone