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2 protocols using diclofenac dfc

1

Evaluation of Progestin Compounds on Cells

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The following active substances were used in the study: the progestin gestobutanoyl (GB) and its metabolite amol were synthesized at the Pirogov Russian National Research Medical University. The structural formulas of GB and amol are shown in Figure 1. The other compounds tested on the cells, medroxyprogesterone acetate (MPA), megestrol acetate (MA), and dienogest (D), as well as the reference substances glucocorticoid hormones dexamethasone (DEX) and hydrocortisone (HC), and the non-steroidal anti-inflammatory drug diclofenac (DFC), were obtained from Sigma (St. Louis, MO, USA).
The following reagents and equipment were used: culture flasks (Eppendorf, Hamburg, Germany); 96-well plates (Corning and Costar, Cambridge, MA, USA); DMEM/F-12 cell culture media with glutamine and RPMI (Gibco, London, UK); fetal bovine serum 10% (Gibco, Auckland, New Zealand); penicillin-streptomycin, trypsin, Versen solution, saline solution, dimethyl sulfoxide (DMSO) (PanEco, Moscow, Russia); amphotericin B (Synthesis AKOMP, Moscow, Russia); 3-(4,5-dimethylthiazol-2-yl)-2,5-tetrazolium bromide (MTT) (Diam, Moscow, Russia); a Uniplan AIFR-01 plate photometer (Ryazan, Russia).
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2

Analytical Workflow for Targeted Micropollutants

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Trimethoprim (TRM), metoprolol (MET), benzotriazole (BTA), carbamazepine (CBZ), caffeine (CAF), propranolol (PRO), sulfamethoxazole (SMZ), furosemide (FRS), mecoprop (MCPP), and diclofenac (DFC) were purchased from Sigma-Aldrich (U.S), and irbesartan (IBT) from TOKYO CHEMICAL INDUSTRY (Japan). Internal standards CAF-D9, SMX-D4, FRS-D5, MCPP-D6, DFC-D4 were purchased from LGC (Germany) and PRO-D7 from MERCK (the Netherlands). Details of these chemicals are presented in Table S1. Hydrogen chloride (HCl) was purchased from VWR chemicals (U.S). Methanol was bought from ACTU-ALL CHEMICALS (the Netherlands). Acetonitrile, ultrawater, and formic acid used in liquid chromatography-mass spectrometry (LC-MS) was purchased from ACTU-ALL CHEMICALS (The Netherlands).
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