title: Whole blood microsampling for the quantitation of estetrol without derivatization by liquid chromatography-tandem mass spectrometry.
abstract: Quantitative bioanalysis and especially pharmacokinetic studies are challenging since only low volumes of biological material are available and low concentrations (ng/ml) are often expected. In this context, volumetric absorptive microsampling (VAMS) devices were developed to accurately collect 10 or 20μl of whole blood from tested subjects. In this study, we present the development and validation of ultra-high performance liquid chromatography coupled to tandem mass spectrometry method after VAMS sampling for the quantitation of estetrol (E4), a potentially new medicine for hormone replacement, contraception and osteoporosis therapies. Interestingly, a very simple sample preparation procedure was developed without any derivatization step. Even if lack of sensitivity is a common consideration when using negative ionization mode, we demonstrated in this work that an excellent sensitivity could be reached by carefully optimizing the nature and concentration of the mobile phase additive. After the optimization of every experimental parameter, the stability, selectivity, trueness, precision and accuracy of the final method were successfully demonstrated. In addition, the excellent performances of the method were confirmed by two independent proof-of-concept pharmacokinetic studies of E4 after VAMS collection in a murine model.
authors.affiliation: Laboratory for the Analysis of Medicines, Center for Interdisciplinary Research on Medicines (CIRM), University of Liège, Quartier hopital, Avenue Hippocrate 15, 4000 Liege, Belgium.
authors.forename: Gwenaël
authors.initials: G
authors.lastname: Nys
authors.affiliation: Laboratory of Tumor and Development Biology, GIGA-Cancer, University of Liège, Quartier hopital, CHU-B23, Avenue Hippocrate 15, 4000 Liege, Belgium.
authors.forename: Anne
authors.initials: A
authors.lastname: Gallez
authors.affiliation: Laboratory for the Analysis of Medicines, Center for Interdisciplinary Research on Medicines (CIRM), University of Liège, Quartier hopital, Avenue Hippocrate 15, 4000 Liege, Belgium.
authors.forename: Miranda G M
authors.initials: MGM
authors.lastname: Kok
authors.affiliation: Laboratory for the Analysis of Medicines, Center for Interdisciplinary Research on Medicines (CIRM), University of Liège, Quartier hopital, Avenue Hippocrate 15, 4000 Liege, Belgium.
authors.forename: Gaël
authors.initials: G
authors.lastname: Cobraiville
authors.affiliation: Laboratory for the Analysis of Medicines, Center for Interdisciplinary Research on Medicines (CIRM), University of Liège, Quartier hopital, Avenue Hippocrate 15, 4000 Liege, Belgium.
authors.forename: Anne-Catherine
authors.initials: AC
authors.lastname: Servais
authors.affiliation: Laboratory of Pharmaceutical Technology and Biopharmacy, CIRM, University of Liège, Quartier hopital, Avenue Hippocrate 15, 4000 Liege, Belgium.
authors.forename: Géraldine
authors.initials: G
authors.lastname: Piel
authors.affiliation: Laboratory of Tumor and Development Biology, GIGA-Cancer, University of Liège, Quartier hopital, CHU-B23, Avenue Hippocrate 15, 4000 Liege, Belgium.
authors.forename: Christel
authors.initials: C
authors.lastname: Pequeux
authors.affiliation: Laboratory for the Analysis of Medicines, Center for Interdisciplinary Research on Medicines (CIRM), University of Liège, Quartier hopital, Avenue Hippocrate 15, 4000 Liege, Belgium. Electronic address: marianne.fillet@ulg.ac.be.
authors.forename: Marianne
authors.initials: M
authors.lastname: Fillet
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D059788:Dried Blood Spot Testing; D004953:Estetrol; D006801:Humans; D051379:Mice; D015203:Reproducibility of Results; D013048:Specimen Handling; D053719:Tandem Mass Spectrometry
chemical_list: D004953:Estetrol
keywords: Ammonium fluoride; Estetrol; Pharmacokinetic study; UHPLC–MS/MS; Volumetric absorptive microsampling