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At regulated levels, these compounds perform vital functions in the body. However, elevated concentrations are related to deregulation of several processes, such as abnormal cell proliferation, which can lead to the development of cancer (breast and ovarian) and other diseases (endometriosis). In this work, an analytical methodology is proposed for the determination of 17\u03b2\u2010estradiol (E1), estrone (E2), and 17\u03b1\u2010ethinylestradiol (EE2) in human urine. Hollow fiber\u2010microporous membrane liquid\u2013liquid extraction (HF\u2010MMLLE) is used with natural deep eutectic solvents (NADESs). This technique was associated with a 96\u2010well plate system followed by high\u2010performance liquid chromatography coupled with a diode array detector (HPLC\u2010DAD). The optimized conditions consisted of using thymol and camphor (1:1 v\/v) as extraction solvent; an extraction time of 100\u00a0min; acetonitrile as desorption solvent; a desorption time of 30\u00a0min; and a sample pH adjusted to 12. Limits of quantification (LOQs) of 25\u00a0ng\u00a0mL\n                    <jats:sup>\u22121<\/jats:sup>\n                    for E2 and 50\u00a0ng\u00a0mL\n                    <jats:sup>\u22121<\/jats:sup>\n                    for E1 and EE2, and limits of detection (LOD) of 7.5\u00a0ng\u00a0mL\n                    <jats:sup>\u22121<\/jats:sup>\n                    for E2 and 15\u00a0ng\u00a0mL\n                    <jats:sup>\u22121<\/jats:sup>\n                    for E1 and EE2 were obtained. Intraday and interday precisions were lower than 8.7% and 21.6%, respectively. Relative recoveries were examined in two samples and ranged from 68.2% to 131.8%. The method was applied in samples from healthy volunteers, and concentrations from 75.6 to 182.7\u00a0ng\u00a0mL\n                    <jats:sup>\u22121<\/jats:sup>\n                    for E2 were found. Finally, the method was evaluated by two sustainable metrics to examine the green aspects of the experimental approach.\n                  <\/jats:p>","DOI":"10.1002\/jssc.70229","type":"journal-article","created":{"date-parts":[[2025,7,18]],"date-time":"2025-07-18T06:56:13Z","timestamp":1752821773000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["A Membrane\u2010Based Strategy for the High\u2010Throughput Determination of Steroidal Hormones in Human Urine Using Natural Deep Eutectic Solvents Combined With Liquid Chromatography Coupled With Diode Array Detector"],"prefix":"10.1002","volume":"48","author":[{"given":"Lucas","family":"Mor\u00e9s","sequence":"first","affiliation":[{"name":"Departamento de Farmacoci\u00eancias Universidade Federal de Ci\u00eancias da Sa\u00fade de Porto Alegre  Porto Alegre Rio Grande do Sul Brazil"},{"name":"Programa de P\u00f3s\u2010Gradua\u00e7\u00e3o em Bioci\u00eancias Universidade Federal de Ci\u00eancias da Sa\u00fade de Porto Alegre  Porto Alegre Rio Grande do Sul Brazil"},{"name":"Departamento de Qu\u00edmica Universidade Federal de Santa Catarina  Florian\u00f3polis Santa Catarina Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Camila","family":"Will","sequence":"additional","affiliation":[{"name":"Departamento de Qu\u00edmica Universidade Federal de Santa Catarina  Florian\u00f3polis Santa Catarina Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eduardo","family":"Carasek","sequence":"additional","affiliation":[{"name":"Departamento de Qu\u00edmica Universidade Federal de Santa Catarina  Florian\u00f3polis Santa Catarina Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Josias","family":"Merib","sequence":"additional","affiliation":[{"name":"Departamento de Farmacoci\u00eancias Universidade Federal de Ci\u00eancias da Sa\u00fade de Porto Alegre  Porto Alegre Rio Grande do Sul 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