{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:08:27Z","timestamp":1760144907283,"version":"build-2065373602"},"reference-count":34,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2024,5,28]],"date-time":"2024-05-28T00:00:00Z","timestamp":1716854400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Mebendazole (MBZ) is a benzimidazole carbamate anthelmintic used worldwide for the treatment and prevention of parasitic disorders in animals and humans. A large number of in vivo and in vitro studies have demonstrated that MBZ also has anticancer activity in multiple types of cancers. After oral administration, the phenylketone moiety of MBZ is rapidly reduced to the hydroxyl group to form the chiral hydroxy metabolite (MBZ-OH). To the best of our knowledge, there is no information in the literature on the stereochemical course of transformation and the anthelmintic and antitumor activity of individual enantiomers of MBZ-OH. In the present study, we describe in detail the direct HPLC resolution of MBZ-OH on a 100 mm \u00d7 4.6 mm Chirapak IG-3 column packed with 3 \u03bcm silica particles containing amylose (3-chloro-5-methylphenylcarbamate) as a selector. At 25 \u00b0C and using pure methanol as the mobile phase, the enantioseparation and resolution factors were 2.38 and 6.13, respectively. These conditions were scaled up at a semi-preparative scale using a 250 mm \u00d7 10 mm Chiralpak IG column to isolate multi-milligram amounts of both enantiomeric forms of the chiral metabolite. The chiroptical properties of the collected enantiomers were determined and, through a theoretical study, were related to the more stable conformations of MBZ-OH. The first and second eluted enantiomers were dextrorotatory and levorotatory, respectively, in dimethylformamide solution. Finally, by recording the retention factors of the enantiomers as the water content in the water\u2013acetonitrile mobile phases was progressively varied, U-shaped retention maps were generated, indicating a dual and competitive hydrophilic interaction liquid chromatography and reversed-phase liquid chromatography retention mechanism on the Chirapak IG-3 chiral stationary phase.<\/jats:p>","DOI":"10.3390\/ph17060696","type":"journal-article","created":{"date-parts":[[2024,5,28]],"date-time":"2024-05-28T05:19:49Z","timestamp":1716873589000},"page":"696","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Chiral Hydroxy Metabolite of Mebendazole: Analytical and Semi-Preparative High-Performance Liquid Chromatography Resolution and Chiroptical Properties"],"prefix":"10.3390","volume":"17","author":[{"given":"Paolo","family":"Guglielmi","sequence":"first","affiliation":[{"name":"Department of Chemistry and Technology of Drugs, Sapienza University of Rome, 00185 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gaia","family":"Pulitelli","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Technology of Drugs, Sapienza University of Rome, 00185 Rome, Italy"},{"name":"National Centre for the Control and Evaluation of Medicines, Chemical Medicines Unit, Istituto Superiore di Sanit\u00e0, Viale Regina Elena 299, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesca","family":"Arrighi","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Technology of Drugs, Sapienza University of Rome, 00185 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniela","family":"Secci","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Technology of Drugs, Sapienza University of Rome, 00185 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4030-8205","authenticated-orcid":false,"given":"Marco","family":"Pierini","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Technology of Drugs, Sapienza University of Rome, 00185 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6346-1953","authenticated-orcid":false,"given":"Roberto","family":"Cirilli","sequence":"additional","affiliation":[{"name":"National Centre for the Control and Evaluation of Medicines, Chemical Medicines Unit, Istituto Superiore di Sanit\u00e0, Viale Regina Elena 299, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,28]]},"reference":[{"key":"ref_1","first-page":"769","article-title":"Anthelmintics A comparative review of their clinical pharmacology","volume":"53","author":"Guyatt","year":"1997","journal-title":"Drugs"},{"key":"ref_2","first-page":"2963","article-title":"Mebendazole elicits a potent antitumor effect on human cancer cell lines both in vitro and in vivo","volume":"8","author":"Mukhopadhyay","year":"2002","journal-title":"Clin. 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