title: Fast and sensitive liquid chromatography-mass spectrometry assay for seven androgenic and progestagenic steroids in human serum.
abstract: A fast and sensitive LC-MS/MS method for the quantitative analysis of seven steroid hormones in 150 μl of human serum was developed and validated. The following compounds were included: 17α-hydroxypregnenolone, 17α-hydroxyprogesterone, androstenedione, dehydroepiandrosterone, testosterone, pregnenolone, and progesterone. Individual stable isotope-labeled analogues were used as internal standards. Sample preparation was performed by liquid-liquid extraction, followed by oxime derivatization to improve the ionization efficiency of the analytes. In contrast to the common derivatization-based methods, the reaction was incorporated into the sample preparation process and the only additional step due to the derivatization was a short heating of the autosampler vials before the sample injection. Chromatographic separation was achieved on a reversed-phase column using a methanol-water gradient. For the analyte detection, a triple quadrupole instrument with electrospray ionization was used. Total run time was 7.0 min and the lower limits of quantification were in the range of 0.03-0.34 nM (0.01-0.10 ng/ml), depending on the analyte. The method was validated using human serum samples from both sexes and applied for the serum steroid profiling of endometriosis patients.
authors.affiliation: Department of Pharmacy, University of Eastern Finland, Yliopistonranta 1 C, FI-70211 Kuopio, Finland. pekka.keski-rahkonen@uef.fi
authors.forename: Pekka
authors.initials: P
authors.lastname: Keski-Rahkonen
authors.forename: Kaisa
authors.initials: K
authors.lastname: Huhtinen
authors.forename: Matti
authors.initials: M
authors.lastname: Poutanen
authors.forename: Seppo
authors.initials: S
authors.lastname: Auriola
mesh_terms: D000728:Androgens / Q000097:blood*; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D011372:Progestins / Q000097:blood*; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000728:Androgens; D011372:Progestins