Simultaneous Green HPLC Determination of Clotrimazole, Miconazole, Metronidazole, and Lidocaine in Pharmaceutical Dosage Forms

Simultaneous Green HPLC Determination of Clotrimazole, Miconazole, Metronidazole, and Lidocaine in Pharmaceutical Dosage Forms | 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 Simultaneous Green HPLC Determination of Clotrimazole, Miconazole, Metronidazole, and Lidocaine in Pharmaceutical Dosage Forms Andrii Zhuravel, Marjan Piponski, Volodymyr Kopilevych, Mamdouh R. Rezk, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9087758/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 15 You are reading this latest preprint version Abstract The analysis of multicomponent pharmaceutical formulations containing drugs with significant diverse physicochemical properties, like 6 orders of magnitude of pKa values, which permanently represents a significant challenge in modern pharmaceutical analysis. The simultaneous determination of multiple active agents offers substantial advantages by reducing analysis time and resource consumption. Furthermore, the increased analytical demand during health emergencies highlights the necessity for validated, rapid, and reliable methods capable of supporting both therapeutic assessment and regulatory decision-making. In the present study, a reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and optimized for the simultaneous determination of four analytes: clotrimazole, miconazole, metronidazole, and lidocaine. Various approaches were investigated for mobile phase composition and column selection to achieve optimal separation and accurate quantitative determination. Chromatographic separation was examined using several columns, including Waters Spherisorb CN (250 × 4.6 mm, 5 µm), LiChrospher 100 CN (125 × 4 mm, 5 µm), LiChrospher 100 CN (250 × 4 mm, 5 µm), Zorbax SB CN (150 × 4.6 mm, 3.5 µm and 250 × 4.6 mm, 5 µm), Zorbax Rx C8 (250 × 4.6 mm, 5 µm), and Zorbax SB Phenyl (150 × 4.6 mm, 3.5 µm). Isocratic elution was employed using mixtures of acetonitrile, methanol, and phosphoric acid or trifluoroacetic acid in different proportions at flow rates of 1.0 and 1.2 mL min⁻¹. UV-detection was carried out at 215 nm. The developed method demonstrated excellent system suitability and complied with ICH Q2 (R2) validation requirements, showing satisfactory linearity (R² > 0.999), accuracy (recoveries within 98–102%), and precision (RSD < 2%). In addition, the sustainability of the proposed method was comprehensively assessed using AGREE, MoGAPI, and BAGI tools to evaluate its greenness and applicability. Overall, the method provides a robust, efficient, and sustainable analytical platform for quality control laboratories, enabling reliable routine analysis of multicomponent pharmaceutical products and therapeutic surveillance. clotrimazole miconazole metronidazole lidocaine HPLC Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 16 Apr, 2026 Reviews received at journal 13 Apr, 2026 Reviews received at journal 12 Apr, 2026 Reviews received at journal 04 Apr, 2026 Reviewers agreed at journal 04 Apr, 2026 Reviews received at journal 03 Apr, 2026 Reviewers agreed at journal 02 Apr, 2026 Reviewers agreed at journal 02 Apr, 2026 Reviewers agreed at journal 02 Apr, 2026 Reviewers agreed at journal 02 Apr, 2026 Reviewers invited by journal 02 Apr, 2026 Editor assigned by journal 02 Apr, 2026 Editor invited by journal 02 Apr, 2026 Submission checks completed at journal 01 Apr, 2026 First submitted to journal 01 Apr, 2026 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. 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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-9087758","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":617689558,"identity":"a9cee1cb-b2ae-4b11-be4b-b5b69c83d54d","order_by":0,"name":"Andrii Zhuravel","email":"","orcid":"","institution":"National University of Life and Environmental Sciences of Ukraine","correspondingAuthor":false,"prefix":"","firstName":"Andrii","middleName":"","lastName":"Zhuravel","suffix":""},{"id":617689559,"identity":"9ad9fb3e-ed1b-4ddc-a298-97ed9b8773ca","order_by":1,"name":"Marjan Piponski","email":"","orcid":"","institution":"Galafarm","correspondingAuthor":false,"prefix":"","firstName":"Marjan","middleName":"","lastName":"Piponski","suffix":""},{"id":617689560,"identity":"8477c870-76cf-4813-914b-cab6a2551b89","order_by":2,"name":"Volodymyr Kopilevych","email":"","orcid":"","institution":"National University of Life and Environmental Sciences of Ukraine","correspondingAuthor":false,"prefix":"","firstName":"Volodymyr","middleName":"","lastName":"Kopilevych","suffix":""},{"id":617689561,"identity":"f44c965a-0a4b-4593-acf2-85be3e1c99df","order_by":3,"name":"Mamdouh R. 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The simultaneous determination of multiple active agents offers substantial advantages by reducing analysis time and resource consumption. Furthermore, the increased analytical demand during health emergencies highlights the necessity for validated, rapid, and reliable methods capable of supporting both therapeutic assessment and regulatory decision-making. In the present study, a reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and optimized for the simultaneous determination of four analytes: clotrimazole, miconazole, metronidazole, and lidocaine. Various approaches were investigated for mobile phase composition and column selection to achieve optimal separation and accurate quantitative determination. Chromatographic separation was examined using several columns, including Waters Spherisorb CN (250 × 4.6 mm, 5 µm), LiChrospher 100 CN (125 × 4 mm, 5 µm), LiChrospher 100 CN (250 × 4 mm, 5 µm), Zorbax SB CN (150 × 4.6 mm, 3.5 µm and 250 × 4.6 mm, 5 µm), Zorbax Rx C8 (250 × 4.6 mm, 5 µm), and Zorbax SB Phenyl (150 × 4.6 mm, 3.5 µm). Isocratic elution was employed using mixtures of acetonitrile, methanol, and phosphoric acid or trifluoroacetic acid in different proportions at flow rates of 1.0 and 1.2 mL min⁻¹. UV-detection was carried out at 215 nm. The developed method demonstrated excellent system suitability and complied with ICH Q2 (R2) validation requirements, showing satisfactory linearity (R² \u003e 0.999), accuracy (recoveries within 98–102%), and precision (RSD \u003c 2%). In addition, the sustainability of the proposed method was comprehensively assessed using AGREE, MoGAPI, and BAGI tools to evaluate its greenness and applicability. Overall, the method provides a robust, efficient, and sustainable analytical platform for quality control laboratories, enabling reliable routine analysis of multicomponent pharmaceutical products and therapeutic surveillance.","manuscriptTitle":"Simultaneous Green HPLC Determination of Clotrimazole, Miconazole, Metronidazole, and Lidocaine in Pharmaceutical Dosage Forms","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-08 16:29:29","doi":"10.21203/rs.3.rs-9087758/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-04-16T12:33:57+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-13T16:10:30+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-12T09:48:45+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-05T03:32:28+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"224847671532548803990353396305344671328","date":"2026-04-04T12:04:37+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-03T04:41:20+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"340205554884941618085852131271011613391","date":"2026-04-03T03:27:47+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"189614411765845427347425100468051652280","date":"2026-04-02T16:25:30+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"34058793116506956881160114796641036739","date":"2026-04-02T14:34:25+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"50217590133173841252769923817956252061","date":"2026-04-02T13:36:30+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-04-02T11:31:34+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-04-02T11:25:50+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2026-04-02T10:06:19+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-04-01T16:57:12+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Chemistry","date":"2026-04-01T16:32:24+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-chemistry","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ccjo","sideBox":"Learn more about [BMC Chemistry](https://bmcchem.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/ccjo/default.aspx","title":"BMC Chemistry","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"3dde2a7d-42aa-425c-aac2-08a8d0716ae9","owner":[],"postedDate":"April 8th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-05-08T11:23:09+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-08 16:29:29","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9087758","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9087758","identity":"rs-9087758","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}