An efficient HPLC workflow for hydrophilic and lipophilic drug combinations on non-conventional polar stationary phase using bio-solvent

An efficient HPLC workflow for hydrophilic and lipophilic drug combinations on non-conventional polar stationary phase using bio-solvent | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article An efficient HPLC workflow for hydrophilic and lipophilic drug combinations on non-conventional polar stationary phase using bio-solvent Mrunalini Kulkarni, Mukesh Parab This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8871395/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Purpose This study presents a novel testing application for co-elution chromatography of biguanide with two different drug classes, sulfonylurea and sodium-glucose co-transporter 2(SGLT2) inhibitors in tablet dosage form. The focus was to quantitate all three drugs in single run on newer chemistry column, additive free mobile phase, using greener solvent and covering wide concentration range. Method This chromatographic method was established on non-conventional C18- embedded polar shielding(EPS) column, an high-purity silica-based reversed-phase column and a simple mobile phase. This method is environmentally responsible, developed with excellent performance for most metrics of greenness indicated by AGREE score 0.73, BAGI score 80.0, AGREE-Prep score 0.73. MoGAPI score 80 and RAPI score 70.0. It showed sustainability and alignment with most of the principles of green analytical chemistry (GAC). A standardized testing protocol was set for estimating three prominent antidiabetic drugs and to overcome difficulty in estimating low-dose Glimepiride and inconsistent regulatory enforcement. Results The method operated at a column flow rate of 0.8 mL min − 1 with a UV detector at 225 nm. The validated method exhibited excellent linearity within 50–150% of the target concentration with correlation coefficients (r) > 0.99 for all analytes. Accuracy studies showed overall mean recoveries of 99.5% for MTH, 100.1% for GPD and 100.2% for CGF. Conclusion The method is precise, accurate, and environmentally responsible, offering a sustainable analytical approach for simultaneous estimation of three drugs in fixed-dose formulations. It exemplifies the sustainable solvent selection and resource-efficient analytical practices integrated to deliver an accessible greener method. HPLC GAC Method development Method validation Metformin HCl Glimepiride and Canagliflozin Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8871395","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":623722606,"identity":"6b25c9c7-98b7-42d6-b200-d58091dc0e7a","order_by":0,"name":"Mrunalini Kulkarni","email":"","orcid":"","institution":"Maharshi Dayanand College of Arts, Science and Commerce","correspondingAuthor":false,"prefix":"","firstName":"Mrunalini","middleName":"","lastName":"Kulkarni","suffix":""},{"id":623722607,"identity":"6361e30a-3b53-44d0-91f7-2caf49dafac2","order_by":1,"name":"Mukesh Parab","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABBklEQVRIiWNgGAWjYJACZiA2YGDgYTzA2MAgBxI58IBILQwgLcZgLQmkaElsAAnh02LOfvbh58Kcw8b8/GcPHPi543D6/LDDD4G22MnpNmDXYtmTbiw9c9thM8kZeQkHe88czt14O80AqCXZ2OwAdi0GB9IYpHm3HbYxuMFjcIC3DahldgJIy4HEbbi0nH/G/Bukxf78GYODf9sOpxvOTv+AX8uNNDaQLWYGDDkGh4G2JMhL5+C3xXLGMzbrmdvSjSVuALXItqUbbpDOKTiQYIDbL+b8acy3C7dZG/b3nzF8+LbNWl5+dvrmDx8q7ORweh9LgGAXx6NFvgG36lEwCkbBKBiZAAA1NGf5DGwJpwAAAABJRU5ErkJggg==","orcid":"","institution":"Maharshi Dayanand College of Arts, Science and Commerce","correspondingAuthor":true,"prefix":"","firstName":"Mukesh","middleName":"","lastName":"Parab","suffix":""}],"badges":[],"createdAt":"2026-02-13 11:38:46","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8871395/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8871395/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":107481796,"identity":"5d4ccbbc-ec84-409c-b495-b726aec9e475","added_by":"auto","created_at":"2026-04-22 02:20:03","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":578107,"visible":true,"origin":"","legend":"","description":"","filename":"AnefficientHPLCworkflowforhydrophiliclipophilicdrugcombinationsonpolarstationaryphase.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8871395/v1_covered_959f7c4b-57e3-4245-8ae5-e86f6ab9b624.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"An efficient HPLC workflow for hydrophilic and lipophilic drug combinations on non-conventional polar stationary phase using bio-solvent","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"HPLC, GAC, Method development, Method validation, Metformin HCl, Glimepiride and Canagliflozin","lastPublishedDoi":"10.21203/rs.3.rs-8871395/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8871395/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003ePurpose\u003c/h2\u003e \u003cp\u003eThis study presents a novel testing application for co-elution chromatography of biguanide with two different drug classes, sulfonylurea and sodium-glucose co-transporter 2(SGLT2) inhibitors in tablet dosage form. The focus was to quantitate all three drugs in single run on newer chemistry column, additive free mobile phase, using greener solvent and covering wide concentration range.\u003c/p\u003e\u003ch2\u003eMethod\u003c/h2\u003e \u003cp\u003eThis chromatographic method was established on non-conventional C18- embedded polar shielding(EPS) column, an high-purity silica-based reversed-phase column and a simple mobile phase. This method is environmentally responsible, developed with excellent performance for most metrics of greenness indicated by AGREE score 0.73, BAGI score 80.0, AGREE-Prep score 0.73. MoGAPI score 80 and RAPI score 70.0. It showed sustainability and alignment with most of the principles of green analytical chemistry (GAC). A standardized testing protocol was set for estimating three prominent antidiabetic drugs and to overcome difficulty in estimating low-dose Glimepiride and inconsistent regulatory enforcement.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eThe method operated at a column flow rate of 0.8 mL min\u0026thinsp;\u0026minus;\u0026thinsp;1 with a UV detector at 225 nm. The validated method exhibited excellent linearity within 50\u0026ndash;150% of the target concentration with correlation coefficients (r)\u0026thinsp;\u0026gt;\u0026thinsp;0.99 for all analytes. Accuracy studies showed overall mean recoveries of 99.5% for MTH, 100.1% for GPD and 100.2% for CGF.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eThe method is precise, accurate, and environmentally responsible, offering a sustainable analytical approach for simultaneous estimation of three drugs in fixed-dose formulations. It exemplifies the sustainable solvent selection and resource-efficient analytical practices integrated to deliver an accessible greener method.\u003c/p\u003e","manuscriptTitle":"An efficient HPLC workflow for hydrophilic and lipophilic drug combinations on non-conventional polar stationary phase using bio-solvent","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-16 22:41:43","doi":"10.21203/rs.3.rs-8871395/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"fefdb980-aaf0-4768-a149-8aee333f944d","owner":[],"postedDate":"April 16th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-05-15T21:38:13+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-16 22:41:43","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8871395","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8871395","identity":"rs-8871395","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}