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.
- 1.Hadley ME, Levine JE (2007) Endocrinology. Pearson Prentice Hall, Upper Saddle River, NJ, USAGoogle Scholar
- 5.Toppari J, Larsen JC, Christiansen P, Giwercman A, Grandjean P, Guillette LJ Jr, Jegou B, Jensen TK, Jouannet P, Keiding N, Leffers H, McLachlan JA, Meyer O, Muller J, Rajpert-De ME, Scheike T, Sharpe R, Sumpter J, Skakkebaek NE (1996) Environ Health Perspect 104(Suppl 4):741–803CrossRefGoogle Scholar
- 8.Gazdar AF, Oie HK, Shackleton CH, Chen TR, Triche TJ, Myers CE, Chrousos GP, Brennan MF, Stein CA, La Rocca RV (1990) Cancer Res 50:5488–5496Google Scholar
- 12.Kaluzny MA, Duncan LA, Merritt MV, Epps DE (1985) J Lipid Res 26:135–140Google Scholar
- 13.Greenland P, Bowley NL, Meiklejohn B, Doane KL, Sparks CE (1990) Clin Chem 36:628–630Google Scholar
- 26.US National Library of Medicine (2010) ChemIDplus Advanced. http://chem.sis.nlm.nih.gov/chemidplus/