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Optimization of SPE-HPLC-FLD methodologies for the extraction and detection of steroid hormones in plasma
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Orientador(es)
Resumo(s)
The quantification of steroid hormones, such as es-tradiol and ethinylestradiol, in plasma requires sensi-tive and reliable analytical methodologies1. This study presents the optimization of a bioanalytical method based on Solid-Phase Extraction (SPE) coupled with High-Performance Liquid Chromatography with Flu-orescence Detection (HPLC-FLD). For sample ex-traction, Strata-X (polymeric reversed-phase) cartridg-es were used, where critical SPE parameters, including cartridge conditioning, matrix washing, and analyte elution, were evaluated. Chromatographic separation was achieved on a C18 stationary phase using a mobile phase of acetonitrile and 0.1% formic acid in ultrapure water. Both isocratic and gradient elution programs were tested to maximize resolution and peak shape. The method was validated for linearity, recovery, and precision (intra- and inter-day), meeting all established criteria for bioanalytical applications. This combined optimization resulted in improved selectivity and sen-sitivity for the determination of estradiol and ethinyl-estradiol in plasma (Figure 1), demonstrating a robust bioanalytical tool for monitoring these compounds in complex biological matrices.
Descrição
NORTE-01-0145-FEDER-000039
Palavras-chave
High-performance liquid chromatography Steroid hormones Solid-phase extraction
Contexto Educativo
Citação
Bracchi, I., Paíga, P., Delerue-Matos, C., Pestana, D., Negrão, R., Keating, E., & Fernandes, V. (2026). Optimization of SPE-HPLC-FLD methodologies for the extraction and detection of steroid hormones in plasma. Book of Abstracts of the 8th Meeting on Medicinal Biotechnology, 29. https://edicoes.ipp.pt/index.php/books/catalog/book/251
Editora
Politema