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A salting\u2010out assisted liquid\u2013liquid extraction optimized by a design of experiments approach was adopted for sample preparations. Isopropanol and magnesium sulfate were the extraction solvent and salting\u2010out agent, respectively. Both drugs and internal standard (terbinafine) were separated using a LiChroCART<jats:sup>\u00ae<\/jats:sup> Purospher Star column (C<jats:sub>18<\/jats:sub>, 55\u00a0\u00d7\u00a04\u00a0mm; 3\u00a0\u03bcm) isocratically pumped with mobile phase [1% triethylamine in water (pH 2.5) and acetonitrile (53:47, v\/v)] at 1\u00a0mL\/min. Stiripentol and terbinafine were detected by fluorescence at 254\/372\u00a0nm and perampanel at 275\/430\u00a0nm. Good linearity was demonstrated for perampanel at 1\u2013500\u00a0ng\/mL range in brain, 2\u20132000\u00a0ng\/mL in liver and 1\u20132000\u00a0ng\/mL in plasma and kidney (<jats:italic>r<\/jats:italic><jats:sup>2<\/jats:sup>\u00a0\u2265\u00a00.9922), and for stiripentol between 10 and 2000\u00a0ng\/mL in brain and 10 and 20\u00a0000\u00a0ng\/mL in the remaining matrices (<jats:italic>r<\/jats:italic><jats:sup>2<\/jats:sup>\u00a0\u2265\u00a00.9917). Precision (CV \u2264\u00a015%) and accuracy (bias \u00b115%) were also verified, with obtained recovery values consistent with those predicted by the experimental design. This method was applied in preliminary pharmacokinetic studies to quantify perampanel or stiripentol after oral administration to mice, showing to be a promising bioanalytical tool to support future nonclinical in vivo pharmacokinetic studies.<\/jats:p>","DOI":"10.1002\/jssc.202000656","type":"journal-article","created":{"date-parts":[[2020,9,30]],"date-time":"2020-09-30T15:37:52Z","timestamp":1601480272000},"page":"4289-4304","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Salting\u2010out assisted liquid\u2013liquid extraction method optimized by design of experiments for the simultaneous high\u2010performance liquid chromatography analysis of perampanel and stiripentol in mouse matrices"],"prefix":"10.1002","volume":"43","author":[{"given":"Sara","family":"Meirinho","sequence":"first","affiliation":[{"name":"CICS\u2010UBI \u2013 Health Sciences Research Center University of Beira Interior  Covilh\u00e3 Portugal"}]},{"given":"Gon\u00e7alo","family":"Campos","sequence":"additional","affiliation":[{"name":"CICS\u2010UBI \u2013 Health Sciences Research Center University of Beira Interior  Covilh\u00e3 Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7010-0619","authenticated-orcid":false,"given":"M\u00e1rcio","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"CICS\u2010UBI \u2013 Health Sciences Research Center University of Beira Interior  Covilh\u00e3 Portugal"},{"name":"UDI\u2010IPG \u2013 Research Unit for Inland Development Polytechnic Institute of Guarda  Guarda Portugal"}]},{"given":"Ana","family":"Fortuna","sequence":"additional","affiliation":[{"name":"Laboratory of Pharmacology Faculty of Pharmacy University of Coimbra  Coimbra Portugal"},{"name":"CIBIT\/ICNAS \u2013 Coimbra Institute for Biomedical Imaging and Translational Research University of Coimbra  Coimbra Portugal"}]},{"given":"Am\u00edlcar","family":"Falc\u00e3o","sequence":"additional","affiliation":[{"name":"Laboratory of Pharmacology Faculty of Pharmacy University of Coimbra  Coimbra Portugal"},{"name":"CIBIT\/ICNAS \u2013 Coimbra Institute for Biomedical Imaging and Translational Research University of Coimbra  Coimbra Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4213-0714","authenticated-orcid":false,"given":"Gilberto","family":"Alves","sequence":"additional","affiliation":[{"name":"CICS\u2010UBI \u2013 Health Sciences Research Center University of Beira Interior  Covilh\u00e3 Portugal"}]}],"member":"311","published-online":{"date-parts":[[2020,10,20]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1080\/14737175.2019.1593142"},{"key":"e_1_2_8_3_1","unstructured":"European Medicines Agency Stiripentol (Diacomit). 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