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3 protocols using etonogestrel

1

Quantitative Analysis of Steroid Hormones

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We purchased etonogestrel, levonorgestrel, medroxyprogesterone acetate, norethindrone, progesterone, testosterone-d3, estradiol, ethinyl estradiol, dansyl chloride, sodium bicarbonate, ammonium acetate and formic acid from Sigma-Aldrich. estradiol-d5 was purchased from CDN Isotopes. progesterone and estradiol stock solutions were purchased from Abbott Laboratories. Water, methanol, acetonitrile, N-butylchloride, and sodium hydroxide were purchased from Fisher Scientific. We purchased double-stripped human serum from Golden West Biologicals.
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2

Etonogestrel and Fluoxetine Preparation

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Etonogestrel (3-ketodesogestrel and fluoxetine were purchased from Sigma-Aldrich (Saint-Quentin Fallavier, France). For in vivo experiments, Etonogestrel was dissolved in oil. For ex vivo experiments, fluoxetine was prepared in saline and for Etonogestrel was dissolved in dimethylsulfoxide with a final concentration dimethylsulfoxide at 0.01%.
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3

Comprehensive LC-MS/MS Steroid Analysis

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Estrone-2,3,4-13C3, BCR576, 17-α-estradiol, E2 (E1024), E1 (E9750 and E1274), desogestrel, etonogestrel, medroxyprogesterone, fulvestrant, tamoxifen, exemestane, and letrozole were from Sigma Aldrich. All estrogen analytes had chemical and isotopic purity greater than 99%. Tam-N-ox, Z-4′Endoxifen, Z-Endoxifen, NDtam, NNDDtam, 4′OHtam, z-α-OHtam, cis-β-OHtam, and 17β-hydroxy exemestane were from Toronto Research Chemicals Inc. Z-4OHtam was from Santa Cruz Biotechnology. E2 (L03801) was from Alfa Aeser. 17-β-Estradiol-2,3,4-13C3, CRM E075 for E1, and CRM-E060 for E2 were from Cerillant. Methanol, ammonium hydroxide, and methyl tert-butyl ether (MTBE) were from VWR. Hexane was from Fischer Scientific.
Steroid-depleted human serum (SDHS) SF236-7 was from BBI Solutions (Crumlin, UK), or was prepared in-house from pooled serum using dextran-coated charcoal. Intralipid was from Fresenius Kabi.
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