{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T21:10:28Z","timestamp":1785186628961,"version":"3.55.0"},"reference-count":39,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2025,2,8]],"date-time":"2025-02-08T00:00:00Z","timestamp":1738972800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004586","name":"FAPERJ","doi-asserted-by":"publisher","award":["210.634\/2024"],"award-info":[{"award-number":["210.634\/2024"]}],"id":[{"id":"10.13039\/501100004586","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004586","name":"FAPERJ","doi-asserted-by":"publisher","award":["E-26\/200.512\/2023"],"award-info":[{"award-number":["E-26\/200.512\/2023"]}],"id":[{"id":"10.13039\/501100004586","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004586","name":"FAPERJ","doi-asserted-by":"publisher","award":["E-26\/211.315\/2021"],"award-info":[{"award-number":["E-26\/211.315\/2021"]}],"id":[{"id":"10.13039\/501100004586","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"CNPq","doi-asserted-by":"publisher","award":["310782\/2022-8"],"award-info":[{"award-number":["310782\/2022-8"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>The Cannabis market is experiencing steady global growth. Cannabis herbal extracts (CHE) are interesting and sought-after products for many clinical conditions. The medical potential of these formulations is mainly attributed to neutral cannabinoids, such as cannabidiol (CBD), tetrahydrocannabinol (THC), and cannabinol (CBN), and their non-standardized content poses a significant fragility in these pharmaceutical inputs. High-resolution mass spectrometry portrays a powerful alternative to their accurate monitoring; however, further analytical steps need to be critically optimized to keep up with instrumental performance. In this study, Full Factorial and Box\u2013Behnken designs were employed to achieve a multivariate optimization of CBD, THC, and CBN extraction from human and veterinary commercial CHE using a minimum methanol\/hexane 9:1 volume and short operational times. A predictive model was also constructed using the Response Surface Methodology and its accuracy was validated. Agitation and sonication times were identified as the most significant operational extraction parameters, followed by the extraction mixture volume. The final setup of the optimized method represented a significantly faster and cheaper protocol than those in the literature. The selected neutral cannabinoids quantification was conducted using ultra high-performance liquid chromatography coupled to high-resolution tandem mass spectrometry (UHPLC-HRMS\/MS) with a precision of &lt;15%, accuracy of 69\u201398%, sensitivity of 23\u201339 ng kg\u22121, and linearity regarding pharmaceutical requirements. State-of-the-art levels of analytical sensitivity and specificity were achieved in the target quantification due to high-resolution mass spectrometry. The developed method demonstrated reliability and feasibility for routine analytical applications. As a proof-of-concept, it enabled the efficient processing of 16 samples of commercial CHE within a three-hour timeframe, showcasing its practicality and reproducibility, and highlighting its potential for broader adoption in similar scenarios for both human and veterinary medicines.<\/jats:p>","DOI":"10.3390\/biom15020246","type":"journal-article","created":{"date-parts":[[2025,2,10]],"date-time":"2025-02-10T05:53:22Z","timestamp":1739166802000},"page":"246","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Optimized Extraction Method for Neutral Cannabinoids Quantification Using UHPLC-HRMS\/MS"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1418-0693","authenticated-orcid":false,"given":"Jo\u00e3o Victor","family":"Meirelles","sequence":"first","affiliation":[{"name":"Departamento de Qu\u00edmica, Pontifi\u0301cia Universidade Cato\u0301lica do Rio de Janeiro, Rio de Janeiro 22451-045, RJ, Brazil"},{"name":"Laborat\u00f3rio Brasileiro de Controle de Dopagem (LBCD), Instituto de Qu\u00edmica, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-909, RJ, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"D\u00e9bora Cristina Diniz","family":"Estevam","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio Brasileiro de Controle de Dopagem (LBCD), Instituto de Qu\u00edmica, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-909, RJ, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8564-0952","authenticated-orcid":false,"given":"Vanessa Farelo","family":"dos Santos","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio Brasileiro de Controle de Dopagem (LBCD), Instituto de Qu\u00edmica, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-909, RJ, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8597-416X","authenticated-orcid":false,"given":"Henrique Marcelo Gualberto","family":"Pereira","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio Brasileiro de Controle de Dopagem (LBCD), Instituto de Qu\u00edmica, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-909, RJ, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7619-5620","authenticated-orcid":false,"given":"Tatiana D.","family":"Saint Pierre","sequence":"additional","affiliation":[{"name":"Departamento de Qu\u00edmica, Pontifi\u0301cia Universidade Cato\u0301lica do Rio de Janeiro, Rio de Janeiro 22451-045, RJ, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1365-7602","authenticated-orcid":false,"given":"Valdir F.","family":"Veiga-Junior","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering, Military Institute of Engineering, Rio de Janeiro 22290-270, RJ, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3760-8231","authenticated-orcid":false,"given":"Monica Costa","family":"Padilha","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio Brasileiro de Controle de Dopagem (LBCD), Instituto de Qu\u00edmica, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-909, RJ, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,8]]},"reference":[{"key":"ref_1","first-page":"17","article-title":"Cannabis in Veterinary Medicine: A Critical Review","volume":"61","author":"Hazzah","year":"2020","journal-title":"AHVMA J."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"253","DOI":"10.3390\/metabo14050253","article-title":"Evaluating the Metabolomic Profile and Anti-Pathogenic Properties of Cannabis Species","volume":"15","author":"Monyela","year":"2024","journal-title":"Metabolites"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jep.2006.02.001","article-title":"Cannabinoids in Medicine: A Review of Their Therapeutic Potential","volume":"105","year":"2006","journal-title":"J. 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