Spectrophotometric determination of the concentrations of pyranine dye components in a ternary mixture for dye batch certification

Spectrophotometric determination of the concentrations of pyranine dye components in a ternary mixture for dye batch certification | 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 Spectrophotometric determination of the concentrations of pyranine dye components in a ternary mixture for dye batch certification Sorina Gabriela Șerban, Maria Laura Strugariu, Adela Diana Berdie, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8873866/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 A strategy is proposed to determine the concentrations of components of pyranine, a cosmetic dye, to batch certification by the U.S. Food and Drug Administration. The main component of pyranine is mainly accompanied by two impurities during its synthesis. Harmful impurities were present at significantly lower concentrations than the main component. Despite the high degree of spectral overlap between the two impurities, specific spectral characteristics were exploited to overcome this challenge and determine their concentrations. The proposed method takes advantage of two key spectral peculiarities of the pyranine system: (1) the main component exhibits significant absorption at 404 nm where the impurities have negligible absorbance, and (2) despite being minor components, show intensely strong UV-vis spectra at concentrations comparable to those obtained in pyranine synthesis, facilitating their detection even at low concentrations. These characteristics enabled the development of a two-stage spectrophotometric approach. First, the concentration of the main dye was determined independently of the presence of the other two dyes in the ternary mixture using a direct absorbance method. Subsequently, the concentrations of the two impurities were determined by bivariate spectral calibration at two selected wavelengths. The results obtained by the proposed method were comparable to those obtained using other methods, such as HPLC and TLC. The proposed method is simple and accurate and can be used in quality control for the certification of dye batches. However, it critically depends on the unique spectral characteristics of the pyranine system, in particular the negligible absorption of the impurities at 404 nm and their relatively high absorption intensity comparable to main dye. UV-Viz spectra Pyranine pyrene dyes direct absorbance method bivariate calibration method 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-8873866","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":592241855,"identity":"9a20d58f-cb2f-45de-82e8-74659ae1f260","order_by":0,"name":"Sorina Gabriela Șerban","email":"","orcid":"","institution":"Politehnica University Timisoara","correspondingAuthor":false,"prefix":"","firstName":"Sorina","middleName":"Gabriela","lastName":"Șerban","suffix":""},{"id":592241856,"identity":"12ab8bba-0656-4e38-8d50-000d0f90b69d","order_by":1,"name":"Maria Laura Strugariu","email":"","orcid":"","institution":"Politehnica University Timisoara","correspondingAuthor":false,"prefix":"","firstName":"Maria","middleName":"Laura","lastName":"Strugariu","suffix":""},{"id":592241857,"identity":"6ec42f18-b5bb-4090-b2e0-1a08d671a390","order_by":2,"name":"Adela Diana Berdie","email":"","orcid":"","institution":"Politehnica University Timisoara","correspondingAuthor":false,"prefix":"","firstName":"Adela","middleName":"Diana","lastName":"Berdie","suffix":""},{"id":592241861,"identity":"e2bd23b6-23b3-4b2e-a959-52b1bf1c81f8","order_by":3,"name":"Simion Jitian","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5klEQVRIiWNgGAWjYDACZgaGAwwMbDwgBsMHIGZjJ1YLD5DBOAOkhZlY23hA2nmghuAF8u3ciQc+7uCTsWdnfvbY5tc2eT5mBsYPH3NwazE4zLvh4MwzIIexmRvn9t02bGNmYJacuQ2PFmbeDYd528B+MZPO7bnNCNTCxsyLR4t8M1wL+zdpy57b9gS1MByGa+Exk2b4cTuRoBaIX0BaDvOUSfY23E5uY2ZsxusX+f6zmz98bDtmz95/fJvEjz+3bee3Nx/88BGfwyDgGIRibAOTDQTVA0ENlP5DjOJRMApGwSgYaQAAAM5Gwhu+WioAAAAASUVORK5CYII=","orcid":"","institution":"Politehnica University Timisoara","correspondingAuthor":true,"prefix":"","firstName":"Simion","middleName":"","lastName":"Jitian","suffix":""}],"badges":[],"createdAt":"2026-02-13 16:23:10","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8873866/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8873866/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":102963315,"identity":"0562ff21-ff62-4e9c-a447-1a8528d871b9","added_by":"auto","created_at":"2026-02-19 04:15:24","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":923587,"visible":true,"origin":"","legend":"","description":"","filename":"Spectrophotometricdeterminationoftheconcentrationsofpyraninedyecomponentsinaternarymixturefordyebatchcertification.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8873866/v1_covered_8ac7e797-9133-4368-98db-1e82eb0eab90.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Spectrophotometric determination of the concentrations of pyranine dye components in a ternary mixture for dye batch certification","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"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":"UV-Viz spectra, Pyranine, pyrene dyes, direct absorbance method, bivariate calibration method","lastPublishedDoi":"10.21203/rs.3.rs-8873866/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8873866/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eA strategy is proposed to determine the concentrations of components of pyranine, a cosmetic dye, to batch certification by the U.S. Food and Drug Administration. The main component of pyranine is mainly accompanied by two impurities during its synthesis. Harmful impurities were present at significantly lower concentrations than the main component. Despite the high degree of spectral overlap between the two impurities, specific spectral characteristics were exploited to overcome this challenge and determine their concentrations. The proposed method takes advantage of two key spectral peculiarities of the pyranine system: (1) the main component exhibits significant absorption at 404 nm where the impurities have negligible absorbance, and (2) despite being minor components, show intensely strong UV-vis spectra at concentrations comparable to those obtained in pyranine synthesis, facilitating their detection even at low concentrations. These characteristics enabled the development of a two-stage spectrophotometric approach. 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