Determination of steroid hormones in blood by GC–MS/MS
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This paper presents the development, optimization and validation of a methodology to determine nine key steroid hormones (viz. pregnenolone, progesterone, dehydroepiandrosterone, androstenedione, testosterone, dihydrotestosterone, estrone, 17α-estradiol and 17β-estradiol) expressed in the steroidogenesis in biological fluids. The analytical method allows for the determination of steroid hormones in blood plasma and serum down to 0.08–0.16 ng/mL for estrogens, 0.20–0.36 ng/mL for androgens and 0.36–0.43 ng/mL for progestagens. These limits of detection were obtainable using a two-step solid-phase clean-up for fractionation and elimination of interfering lipids (fatty acids, phospholipids, glycerides and sterols) from the steroid hormones. The accuracy of the method was 50–112% in the range 0.10 to 2.00 ng/mL.
KeywordsPlasma Serum Solid-phase extraction Gas chromatography Lipid separation Biological samples
This work was supported by the Pathos Centre (ENV 2104-07-0015) funded by the Danish Strategic Research Council, the Drug Research Academy (Faculty of Pharmaceutical Sciences, University of Copenhagen) and a climate-change project funded by Dean Sven Frøkjær, Faculty of Pharmaceutical Sciences, University of Copenhagen. Kenneth M. Pedersen is gratefully acknowledged for his technical assistance in our EnvTox research group.
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