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Following the definition of the analytical target profile (ATP), encompassing the critical analytical attributes (CAA), a two-level risk assessment strategy (Ishikawa diagram\u2014failure mode and effect analysis (FMEA)) was employed to identify the critical method parameters (CMPs) with an extensive impact on method performance. The behavior of the CMPs (flow rate and mobile phase composition) was further characterized by experimental design, resorting to a face-centered central composite design (FcCCD). Statistical modeling, response surface analysis, and Monte Carlo simulations led to the definition of the Method Operable Design Region (MODR), associated with a negligible risk of failing the predefined CAA specifications. The optimal method was validated according to international regulatory recommendations. Apart from guaranteeing linearity, accuracy, precision, specificity, robustness, and stability, these conditions were found to be suitable for analysis using a different HPLC column and equipment. In a nutshell, the development and optimization strategies, under the comprehensive framework of AQbD, provided an effective, simple, rapid, reliable, and flexible method for routine analysis of the compounds in research or industrial environments.<\/jats:p>","DOI":"10.3390\/chemosensors9070172","type":"journal-article","created":{"date-parts":[[2021,7,6]],"date-time":"2021-07-06T23:53:51Z","timestamp":1625615631000},"page":"172","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Expediting Disulfiram Assays through a Systematic Analytical Quality by Design Approach"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3212-7324","authenticated-orcid":false,"given":"Jo\u00e3o","family":"Basso","sequence":"first","affiliation":[{"name":"Faculty of Pharmacy, University of Coimbra, 3000-548 Coimbra, Portugal"},{"name":"Coimbra Chemistry Centre, Department of Chemistry, University of Coimbra, 3004-504 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3230-8045","authenticated-orcid":false,"given":"Maria Lu\u00edsa","family":"Ramos","sequence":"additional","affiliation":[{"name":"Coimbra Chemistry Centre, Department of Chemistry, University of Coimbra, 3004-504 Coimbra, Portugal"}]},{"given":"Alberto","family":"Pais","sequence":"additional","affiliation":[{"name":"Coimbra Chemistry Centre, Department of Chemistry, University of Coimbra, 3004-504 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3636-5805","authenticated-orcid":false,"given":"Rui","family":"Vitorino","sequence":"additional","affiliation":[{"name":"iBiMED\u2014Department of Medical Sciences, University of Aveiro, 3810-193 Aveiro, Portugal"},{"name":"UnIC\u2014Cardiovascular Research & Development Centre, Department of Surgery and Physiology, Faculty of Medicine, University of Porto, 4200-319 Porto, Portugal"},{"name":"QOPNA & LAQV-REQUIMTE, Chemistry Department, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"given":"Ana","family":"Fortuna","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy, University of Coimbra, 3000-548 Coimbra, Portugal"},{"name":"Coimbra Institute for Biomedical Imaging and Translational Research, University of Coimbra, 3000-548 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3424-548X","authenticated-orcid":false,"given":"Carla","family":"Vitorino","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy, University of Coimbra, 3000-548 Coimbra, Portugal"},{"name":"Coimbra Chemistry Centre, Department of Chemistry, University of Coimbra, 3004-504 Coimbra, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,7,6]]},"reference":[{"key":"ref_1","unstructured":"Brunton, L.L., Hilal-Dandan, R., and Knollmann, B.C. 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