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2 protocols using 13c3 estrone

1

Comprehensive Steroid Analysis Workflow

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The internal standards (d3-testosterone, d4-cortisol, d9-progesterone, d7-cholesterol, 13c3-androstene-3,17-dione, and 13c3-estrone) were purchased from Cerilliant (Round Rock, TX, USA). L-ascorbic acid, sodium acetate, acetic acid, β-glucosaldosidase/arylsulfatase, ammonium iodide (NH4I), dithioerythritol (DTE), and N-methyl-n-(trimethylsilyl)trifluoroacetamide (MSTFA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Commercial standards of steroids (dehydroepiandrosterone, testosterone, 11-deoxycorticosterone, cortisone, cortisol, 16α-hydroxyestrone, pregnenolone, 17α-hydroxypregnenolone, epipregnanolone, dihydrotestosterone, 21-hydroxyprogesterone, androsterone, epiandrosterone, corticosterone, estrone, 17β-estradiol, estriol, 16-epiestriol, progesterone, 17α-hydroxyprogesterone, desmosterol, cholesterol, etc.) were purchased from Sigma-Aldrich, J&K Chemical Ltd. (Beijing, China), or Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). Chromatographic pure hexane, methanol, and ethyl acetate were purchased from Merck (Fairfield, OH, USA). The Oasis HLB SPE cartridge was obtained from Waters (1.5 ml, 60 mg; Waters, Milford, MA, USA).
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2

LC-MS Analysis of Steroid Hormones

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Powder of E3S sodium salt (Sigma-Aldrich, St. Louis, MO, USA) dissolved in methanol, solutions of 17β-estradiol (LGC Standards, Luckenwalde, Germany), estrone and progesterone (Cerilliant, Round Rock, TX, USA) were used as reference standard. D 4 -estrone-3-sulfate sodium salt (Sigma-Aldrich, St. Louis, MO, USA) dissolved in methanol (stock solution), solutions of 13 C 3 -estrone, d 5 -17β-estradiol and d 9 -progesterone (Cerilliant, Round Rock, TX, USA) were used as an internal standard. Water, acetonitrile and methanol (LC-MS grade) were purchased from Biosolve (Biosolve, Dieuze, France). Ammonia solution (25% in water) (Merck KGaA, Darmstadt, Germany) was Suprapur® grade and was purchased from Sigma-Aldrich (St. Louis, MO, USA). Bovine serum albumin (BSA), dansyl chloride and carbonatebicarbonate buffer (0.05 M, pH = 10.5) used for the derivatization were purchased from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO, USA).
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