{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T06:38:19Z","timestamp":1792737499296,"version":"4.5.0"},"reference-count":42,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T00:00:00Z","timestamp":1792627200000},"content-version":"vor","delay-in-days":21,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Separation Science Plus"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Citrus flavonoids are well known for their health\u2010promoting effects and contribution to daily nutritional dietary supplements. Hesperetin (HST), a citrus flavonoid, has gained special attention due to its multiple pharmacological actions, including anticancer, antioxidant, and anti\u2010inflammatory effects. However, identification of degradation products (DPs) is a key concern for preclinical development of HST. To achieve this, stability studies are essential to establish appropriate storage conditions and optimize the formulations accordingly. Despite this, substantial chemical stability data for HST applying RP\u2010HPLC, LC\u2010ESI\u2010MS\/MS, and NMR, along with in vitro cytotoxicity, have not yet been investigated. To address these gaps, the impact of different stress conditions, namely hydrolytic (basic, acidic, and neutral), oxidative, thermal, and light exposure, was tested on HST. HST and its DPs were separated using a developed RP\u2010HPLC\u2010based stability\u2010indicating method. The developed method revealed that HST was highly stable under all stress conditions except basic conditions, where three DPs were formed. Out of the three DPs, two major DPs were isolated and enriched using mass\u2010triggered LC and subsequently characterized thoroughly by LC\u2010ESI\u2010MS\/MS, and 1D\/2D NMR spectroscopy. The DPs were identified as DP1 [(E)\u20103\u2010(3\u2010hydroxy\u20104\u2010methoxyphenyl) acrylic acid] and DP2 [(E)\u20105\u2010(3\u2010hydroxy\u20104\u2010methoxystyryl) benzene\u20101,3\u2010diol], which are novel and have not been reported in the literature to date. The isolated DPs were evaluated for cytotoxicity using an invitro MTT assay on Dalton's lymphoma (DL) cancer cell line and peripheral blood mononuclear cells, along with in silico absorption, distribution, metabolism, excretion, and toxicity profiling. Both DPs induce minimal cytotoxicity in healthy cells, with cell death remaining below 10% throughout the treatment period. Additionally, Greenness assessment using the Analytical Greenness and Analytical Green Star Area tools indicated an acceptable environmental impact. The developed method effectively assessed HST stability under stressed conditions, supporting its use in routine quality control and formulation development. The study also offers valuable insights for the preclinical development of HST as a novel anticancer agent.<\/jats:p>","DOI":"10.1002\/sscp.70310","type":"journal-article","created":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T05:55:42Z","timestamp":1792734942000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Impact of Stress Conditions on Hesperetin Stability: Isolation and Structural Characterization of Major Degradation Products Using Dual\u2010Mode Prep\u2010LC\u2010PDA\/MS, LC\u2010MS\/MS, 1D\/2D NMR, In Vitro Cytotoxicity and Greenness Assessment"],"prefix":"10.1002","volume":"9","author":[{"given":"Prajkta","family":"Bondarkar","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Analysis National Institute of Pharmaceutical Education and Research Guwahati (NIPER\u2010G)  Kamrup Assam India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pintu","family":"Das","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry National Institute of Pharmaceutical Education and Research Guwahati (NIPER\u2010G)  Kamrup Assam India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-0500-6053","authenticated-orcid":false,"given":"Pratik Vilas","family":"Gore","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Analysis National Institute of Pharmaceutical Education and Research Guwahati (NIPER\u2010G)  Kamrup Assam India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-3750-2352","authenticated-orcid":false,"given":"Dnyaneshwar","family":"Maykar","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Analysis National Institute of Pharmaceutical Education and Research Guwahati (NIPER\u2010G)  Kamrup Assam India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7336-0408","authenticated-orcid":false,"given":"Sandeep","family":"Jat","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Analysis National Institute of Pharmaceutical Education and Research Guwahati (NIPER\u2010G)  Kamrup Assam India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-8539-8692","authenticated-orcid":false,"given":"Saddam","family":"Hussain","sequence":"additional","affiliation":[{"name":"Department of Zoology Cell and Biochemical Technology Laboratory Cotton University  Guwahati Assam India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8176-5694","authenticated-orcid":false,"given":"Akalesh Kumar","family":"Verma","sequence":"additional","affiliation":[{"name":"Department of Zoology Cell and Biochemical Technology Laboratory Cotton University  Guwahati Assam India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Veena","family":"K. 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