{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,15]],"date-time":"2025-12-15T08:29:04Z","timestamp":1765787344637,"version":"3.48.0"},"reference-count":41,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T00:00:00Z","timestamp":1764633600000},"content-version":"vor","delay-in-days":1,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Chirality"],"published-print":{"date-parts":[[2025,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Odevixibat, a selective and reversible inhibitor of the ileal bile acid transporter (IBAT), received approval from the US Food and Drug Administration (FDA) in 2021 as the first therapeutic option for pruritus associated with progressive familial intrahepatic cholestasis (PFIC). In this study, a comprehensive chiral high\u2010performance liquid chromatography (HPLC) method was developed for the enantiomeric and diastereomeric separation of Odevixibat and its stereoisomeric impurities (RS, RR, SS, and SR) using immobilized polysaccharide\u2010based chiral stationary phases. The various chromatographic modes\u2014including normal\u2010phase (NP), reversed\u2010phase (RP), and polar organic (PO)\u2014were systematically investigated. Optimal resolution was achieved using a CHIRALPAK IA column (4.6\u2009\u00d7\u2009250\u2009mm, 3\u2009\u03bcm) with a mobile phase of\n                    <jats:italic>n<\/jats:italic>\n                    \u2010hexane\/isopropanol\/methanol\/trifluoroacetic acid (50:35:15:0.1, v\/v\/v\/v) at 40\u00b0C and a flow rate of 1.0\u2009mL\/min. The method was validated according to ICH Q2(R1) guidelines. Additionally, the CHIRALPAK IM in NP mode effectively resolved closely eluting isomeric impurities, enabling preparative isolation and characterization. The optimized RP\u2010mode method was compatible with LC\u2013MS detection, facilitating trace\u2010level impurity profiling on CHIRALPAK ID\u20103 and structural characterization of stereoisomers and process\u2010related impurities. Thermodynamic analysis revealed entropy\u2010driven enantioseparations, with the NP\u2010HPLC method on CHIRALPAK IA showing more favorable interactions (\u0394G\u00b0\u2009=\u20090.06) than the RP\u2010HPLC system (\u0394G\u00b0\u2009=\u20091.34). The method demonstrated excellent linearity (\n                    <jats:italic>r<\/jats:italic>\n                    <jats:sup>2<\/jats:sup>\n                    \u2009&gt;\u20090.999) over the concentration range of 0.028\u20130.211\u2009\u03bcg\/mL, with recoveries between 87.1%\u2013101.2% and precision within 0.8% RSD. High sensitivity was achieved, with LODs of 0.009\u20130.025\u2009\u03bcg\/mL and LOQs of 0.028\u20130.075\u2009\u03bcg\/mL. The method proved robust under deliberate variations in column temperature (\u00b11\u00b0C), mobile phase composition (\u00b12%), and flow rate (1.0\u2009\u00b1\u20090.1\u2009mL\u2009min\n                    <jats:sup>\u22121<\/jats:sup>\n                    ), showing no significant change in enantioresolution. These complementary approaches are suitable for routine quality control (CHIRALPAK IA), purification (CHIRALPAK IM) process monitoring and LC\u2013MS impurity profiling (CHIRALPAK ID\u20103), and regulatory submissions, supporting current good manufacturing practices (cGMP) in the development of Odevixibat.\n                  <\/jats:p>","DOI":"10.1002\/chir.70065","type":"journal-article","created":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T09:55:35Z","timestamp":1764669335000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Enantiomeric Resolution of Odevixibat via Immobilized Polysaccharide Columns: Impact of NP, RP, and PO Modes on LC\u2013MS\u2010Compatible Method Design"],"prefix":"10.1002","volume":"37","author":[{"given":"S.\u00a0R.\u00a0Krishna\u00a0Murthy","family":"Kasa","sequence":"first","affiliation":[{"name":"Department of Chemistry, GITAM School of Science GITAM (Deemed to be) University  Hyderabad India"},{"name":"Daicel Chiral Technologies (India) Pvt Ltd., IKP Knowledge Park  Hyderabad India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7772-3173","authenticated-orcid":false,"given":"Muvvala","family":"Venkatanarayana","sequence":"additional","affiliation":[{"name":"Department of Chemistry, GITAM School of Science GITAM (Deemed to be) University  Hyderabad India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lakshmi\u00a0Narayana","family":"Chennuru","sequence":"additional","affiliation":[{"name":"Daicel Chiral Technologies (India) Pvt Ltd., IKP Knowledge Park  Hyderabad India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2399-770X","authenticated-orcid":false,"given":"M.\u00a0V.\u00a0N.\u00a0Kumar","family":"Talluri","sequence":"additional","affiliation":[{"name":"Daicel Chiral Technologies (India) Pvt Ltd., IKP Knowledge Park  Hyderabad India"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,12,2]]},"reference":[{"volume-title":"Odevixibat Prescribing Information","year":"2021","author":"Albireo Pharma Inc","key":"e_1_2_7_2_1"},{"key":"e_1_2_7_3_1","first-page":"390","article-title":"Odevixibat in PFIC: A Phase 3 Trial","volume":"7","author":"Thompson R. 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Y.","year":"2000","journal-title":"Chromatographia"},{"key":"e_1_2_7_12_1","first-page":"152","article-title":"Chirality and Environmental Pollution","volume":"84","author":"Ali I.","year":"2003","journal-title":"Current Science"},{"key":"e_1_2_7_13_1","first-page":"2863","article-title":"Chromatographic and Electrophoretic Enantioseparation of Chiral Quinolones: Physicochemical and Pharmaceutical Properties","volume":"40","author":"Zhang Y.","year":"2017","journal-title":"Journal of Separation Science"},{"key":"e_1_2_7_14_1","first-page":"789","article-title":"Comparative Study of the Chiral Resolution of Econazole, Miconazole, and Sulconazole Using Amylose\u2010Based Chiral Stationary Phases Under Normal\u2010Phase Conditions","volume":"54","author":"Ali I.","year":"2001","journal-title":"Chromatographia"},{"issue":"3","key":"e_1_2_7_15_1","article-title":"Method Development and Validation of Odevixibat by HPLC and Characterization of Its Novel Stress Degradation Products by HPLC\u2013MS\/MS","volume":"27","author":"Sudheer C.","year":"2024","journal-title":"African Journal of Biomedical Research"},{"key":"e_1_2_7_16_1","article-title":"Comparison of Chiral LC Modes Using Protein\u2010 and Polysaccharide\u2010Based CSPs","volume":"1624","author":"Dob\u00f3 M.","year":"2020","journal-title":"Journal of Chromatography B"},{"key":"e_1_2_7_17_1","first-page":"1123","article-title":"Enantioselective RP\u2010UHPLC\u2013MS\/MS Using Immobilized CSPs","volume":"44","author":"Karongo T.","year":"2021","journal-title":"Journal of Separation Science"},{"key":"e_1_2_7_18_1","article-title":"Recent Advances in CSPs for Chiral HPLC","volume":"143","author":"Zhang T.","year":"2021","journal-title":"TrAC Trends in Analytical Chemistry"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/9781118592014"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1521-3773(19980504)37:8<1020::AID-ANIE1020>3.0.CO;2-5"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9673(00)00951-1"},{"key":"e_1_2_7_22_1","first-page":"3","article-title":"Influence of Mobile Phase Composition on Enantioselectivity of Polysaccharide\u2010Based CSPs","volume":"875","author":"Lindner W.","year":"2000","journal-title":"Journal of Chromatography A"},{"key":"e_1_2_7_23_1","first-page":"10","article-title":"Optimization of Preparative Chromatographic Processes: A Practical Guide","volume":"1505","author":"Pellegatti M.","year":"2017","journal-title":"Journal of Chromatography A"},{"key":"e_1_2_7_24_1","first-page":"896","article-title":"Chiral Separations: The Business of Making Mirrors","volume":"1216","author":"Welch C. 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K.","year":"2022","journal-title":"Journal of Pharmaceutical and Biomedical Analysis"},{"key":"e_1_2_7_29_1","first-page":"1340","article-title":"Recent Trends in the Application of Reversed\u2010Phase Chromatography for Enantiomeric Separations","volume":"42","author":"Rossi M.","year":"2021","journal-title":"Electrophoresis"},{"key":"e_1_2_7_30_1","article-title":"Enhanced Enantiomeric Resolution and MS Compatibility Using Reversed\u2010Phase Chiral LC With Polysaccharide\u2010Based CSPs","volume":"36","author":"Zhang T.","year":"2022","journal-title":"Rapid Communications in Mass Spectrometry"},{"key":"e_1_2_7_31_1","article-title":"Enantioselective Determination of \u03b2\u2010Blockers in Biological Fluids by Reversed\u2010Phase Chiral LC\u2013MS\/MS","volume":"1646","author":"Zhou H.","year":"2021","journal-title":"Journal of Chromatography A"},{"key":"e_1_2_7_32_1","article-title":"Polysaccharide\u2010Based Chiral Stationary Phases: Recent Advances in Enantioselective Liquid Chromatography","volume":"1627","author":"Franco P.","year":"2020","journal-title":"Journal of Chromatography A"},{"key":"e_1_2_7_33_1","first-page":"108","article-title":"Advances in Immobilized Polysaccharide CSPs for LC and SFC","volume":"42","author":"Yamamoto C.","year":"2019","journal-title":"Journal of Separation Science"},{"key":"e_1_2_7_34_1","article-title":"Application of Reversed\u2010Phase Chiral LC\u2013MS for the Identification of Chiral Metabolites in Drug Metabolism Studies","volume":"180","author":"Wang J.","year":"2020","journal-title":"Journal of Pharmaceutical and Biomedical Analysis"},{"key":"e_1_2_7_35_1","first-page":"33","article-title":"Identification of Stereoisomers of Pharmaceutical Compounds Using Reversed\u2010Phase Chiral LC\u2013MS","volume":"1566","author":"West C.","year":"2018","journal-title":"Journal of Chromatography A"},{"volume-title":"International Council for Harmonisation (ICH). 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