{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:34:47Z","timestamp":1789691687279,"version":"4.0.1"},"reference-count":23,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2022,12,22]],"date-time":"2022-12-22T00:00:00Z","timestamp":1671667200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Deputyship for Research and Innovation \u201cMinistry of Education\u201d in Saudi Arabia","award":["IFKSURG-754"],"award-info":[{"award-number":["IFKSURG-754"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>The combination regimen targeting BRAF and MEK inhibition, for instance, encorafenib (Braftovi\u2122, ENF) plus binimetinib (Mektovi\u00ae, BNB), are now recommended as first-line treatment in patients with unresectable or metastatic melanoma with a BRAF V600-activating mutation. Patients treated with combination therapy of ENF and BNB demonstrated a delay in resistance development, increases in antitumor activity, and attenuation of toxicities compared with the activity of either agent alone. However, the pharmacokinetic profile of the FDA-approved ENF and BNB is still unclear. In this study, a rapid and sensitive LC-MS\/MS bioanalytical method for simultaneous quantification of ENF and BNB in rat plasma was developed and validated. Chromatography was performed on an Agilent Eclipse plus C18 column (50 mm \u00d7 2.1 mm, 1.8 \u00b5m), with an isocratic mobile phase composed of 0.1% formic acid in water\/acetonitrile (67:33, v\/v, pH 3.2) at a flow rate of 0.35 mL\/min. A positive multiple reaction monitoring (MRM) mode was chosen for detection and the process of analysis was run for 2 min. Plasma samples were pre-treated using protein precipitation with acetonitrile containing spebrutinib as the internal standard (IS). Method validation was assessed as per the FDA guidelines for the determination of ENF and BNB over concentration ranges of 0.5\u20133000 ng\/mL (r2 \u2265 0.997) for each drug (plasma). The lower limits of detection (LLOD) for both drugs were 0.2 ng\/mL. The mean relative standard deviation (RSD) of the results for accuracy and precision was \u2264 7.52%, and the overall recoveries of ENF and BNB from rat plasma were in the range of 92.88\u2013102.28%. The newly developed approach is the first LC\u2013MS\/MS bioanalytical method that can perform simultaneous quantification of ENF and BNB in rat plasma and its application to a pharmacokinetic study. The mean result for Cmax for BNB and ENF was found to be 3.43 \u00b1 0.46 and 16.42 \u00b1 1.47 \u00b5g\/mL achieved at 1.0 h for both drugs, respectively. The AUC0-\u221e for BNB and ENF was found to be 18.16 \u00b1 1.31 and 36.52 \u00b1 3.92 \u00b5g\/mL.h, respectively. On the other hand, the elimination half-life (t1\/2kel) parameters for BNB and ENF in the rat plasma were found to be 3.39 \u00b1 0.43 h and 2.48 \u00b1 0.24 h, and these results are consistent with previously reported values.<\/jats:p>","DOI":"10.3390\/molecules28010079","type":"journal-article","created":{"date-parts":[[2022,12,23]],"date-time":"2022-12-23T03:26:25Z","timestamp":1671765985000},"page":"79","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["A Rapid and Sensitive Liquid Chromatography-Tandem Mass Spectrometry Bioanalytical Method for the Quantification of Encorafenib and Binimetinib as a First-Line Treatment for Advanced (Unresectable or Metastatic) Melanoma\u2014Application to a Pharmacokinetic Study"],"prefix":"10.3390","volume":"28","author":[{"given":"Mohamed M.","family":"Hefnawy","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia"},{"name":"Department of Analytical Chemistry, Faculty of Pharmacy, Mansoura University, Mansoura 35516, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0483-8113","authenticated-orcid":false,"given":"Mohammed M.","family":"Alanazi","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abdullah M.","family":"Al-Hossaini","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1197-8672","authenticated-orcid":false,"given":"Abdulaziz I.","family":"Alnasser","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7197-1515","authenticated-orcid":false,"given":"Adel S.","family":"El-Azab","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3302-4036","authenticated-orcid":false,"given":"Yousef A. Bin","family":"Jardan","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1147-4960","authenticated-orcid":false,"given":"Mohamed W.","family":"Attwa","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manal A.","family":"El-Gendy","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"7","DOI":"10.3322\/caac.21708","article-title":"Cancer Statistics, 2022","volume":"72","author":"Siegel","year":"2022","journal-title":"CA Cancer J. 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