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• W2022804323 abstract "Oestrogens, such as oestrone (E1), 17β-oestradiol (E2), oestriol (E3) and their biologically active metabolites 2-methoxyoestrone (2-MeOE1), 2-hydroxyoestradiol (2-OHE2) 16-ketooestradiol (16-OE2), 16-epioestriol (16-epiE3), as well as testosterone (T) play an important role in physiological and pathological developmental processes during human development. We therefore aimed at developing an isotope dilution/bench top gas chromatography–mass spectrometry (ID/GC–MS) method, based on benchtop GC–MS, for the simultaneous determination (‘profiling’) of the above analytes in children. The method consisted of equilibration of urine (5 ml) with a cocktail containing stable isotope-labelled analogues of the analytes as internal standards ([2,4-2H2]E1, [2,4,16,16-2H4]E2, [2,4,17-2H3]E3, [16,16,17-2H3]T, [1,4,16,16-2H4]2-MeOE1, [1,4,16,16,17-2H5]2-OHE2, [2,4,15,15,17-2H5]16-OE2 and [2,4-2H2]16-epiE3). Then, solid-phase extraction (C18 cartridges), enzymatic hydrolysis (sulphatase from Helix pomatia (type H-1)), re-extraction, purification by anion exchange chromatography and derivatisation to trimethylsilyl ethers followed. The samples were analysed by GC–MS (Agilent GC 6890N/5975MSD; fused silica capillary column 25 m × 0.2 mm i.d., film 0.10 μm). Calibration plots were linear and showed excellent reproducibility with coefficients of determination (r2) between 0.999 and 1.000. Intra- and inter-assay coefficients of variation (CV) were <2.21% for all quantified metabolites. Sensitivity was highest for 2-OHE2 (0.25 pg per absolute injection: signal-to-noise ratio (S/N) = 3) and lowest for 16-epiE3 (2 pg per absolute injection: S/N = 2.6), translating into corresponding urine sample analyte concentrations of 0.025 ng ml−1 and 0.2 ng ml−1, respectively. Accuracy – determined in a two-level spike experiment – showed relative errors ranging between 0.15% for 16-OE2 and 11.63% for 2-OHE2. Chromatography showed clear peak shapes for the components analysed. In summary, we describe a practical, sensitive and specific ID/GC–MS assay capable of profiling the above-mentioned steroids in human urine from childhood onwards." @default.
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• W2022804323 date "2010-12-01" @default.
• W2022804323 modified "2023-10-17" @default.
• W2022804323 title "Profiling oestrogens and testosterone in human urine by stable isotope dilution/benchtop gas chromatography–mass spectrometry" @default.
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• W2022804323 doi "https://doi.org/10.1016/j.steroids.2010.06.014" @default.
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