A High Performance Thin Layer Chromatography Method by Design of Experiment approach for quantification of Ascorbic acid in Extracts of Nycthanthus Arbortristis (Parijat) Leaves

A High Performance Thin Layer Chromatography Method by Design of Experiment approach for quantification of Ascorbic acid in Extracts of Nycthanthus Arbortristis (Parijat) Leaves | 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 A High Performance Thin Layer Chromatography Method by Design of Experiment approach for quantification of Ascorbic acid in Extracts of Nycthanthus Arbortristis (Parijat) Leaves Pooja Deshpande, S. J. Daharwal, Nagendra Singh Chauhan, Umakant Sahu, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9539312/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 The present study was executed for method development, validation and to quantify Ascorbic acid in Parijat leaves (In- house formulation) and compared with marketed herbal formulations of parijat leaves by high-performance thin-layer chromatography (HPTLC). Isolation of bioactive compounds was carried out using the mobile phase, Methanol: Chloroform : Ammonia (5: 4: 1) and detected at wavelength 273 nm. This method was validated in terms of Accuracy, Precision, Linearity, Limit of detection (LOD) and Limit of quantification (LOQ). The calibration range was found to be 0.5– 4.5 ng/spot for the active compound. Ascorbic acid was separated with an Rf value 0.55. The method was validated for linearity (0.5–4.5 ng/spot), accuracy (97–99%), and RSD of precision (0.13–1.20%). The LOD and LOQ were found to be 0.0363 ng/spot and 0.1099 ng/spot respectively. The quantification of ascorbic acid in various formulations was in the range of (357.1 µg to 283.8 µg). The design of experiment software was applied by taking three independent variables such as the volume of solvent (methanol), solvent system, and chamber saturation time were investigated for analytical quality by design. This developed HPTLC method is simple, precise, robust, specific, rapid, and cost effective. It could be used for quality control study and quantification of ascorbic acid in the plant and different herbal formulations containing the plant species. Method development Quantification Validation Herbal formulations 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-9539312","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":634393320,"identity":"ec545d25-7492-4268-aeeb-74555669c0b7","order_by":0,"name":"Pooja Deshpande","email":"","orcid":"","institution":"Mody University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Pooja","middleName":"","lastName":"Deshpande","suffix":""},{"id":634393323,"identity":"dde342f2-74ff-4e6c-bcff-9948ca3379e0","order_by":1,"name":"S. J. Daharwal","email":"","orcid":"","institution":"University Institute of Pharmacy, Pandit Ravishankar Shukla University","correspondingAuthor":false,"prefix":"","firstName":"S.","middleName":"J.","lastName":"Daharwal","suffix":""},{"id":634393327,"identity":"c6758c75-e080-4aea-9b69-55957514845b","order_by":2,"name":"Nagendra Singh Chauhan","email":"","orcid":"","institution":"drug testing laboratory Avam Anusandhan Kendra (Ayush)","correspondingAuthor":false,"prefix":"","firstName":"Nagendra","middleName":"Singh","lastName":"Chauhan","suffix":""},{"id":634393330,"identity":"d5402997-6fdf-4c38-8494-8afc2721218e","order_by":3,"name":"Umakant Sahu","email":"","orcid":"","institution":"University Institute of Pharmacy, Pandit Ravishankar Shukla University","correspondingAuthor":false,"prefix":"","firstName":"Umakant","middleName":"","lastName":"Sahu","suffix":""},{"id":634393335,"identity":"0d92094d-2e9d-4848-9a62-fc558c6ba282","order_by":4,"name":"Khushbu Singh","email":"","orcid":"","institution":"Mahakal Institute of Pharmaceutical sciences, Sam Higginbottom University of agriculture, Technology and Sciences","correspondingAuthor":false,"prefix":"","firstName":"Khushbu","middleName":"","lastName":"Singh","suffix":""},{"id":634393340,"identity":"6c097a05-e5d1-44e4-aeaf-4faeaef8168a","order_by":5,"name":"Deenanath Jhade","email":"","orcid":"","institution":"Mody University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Deenanath","middleName":"","lastName":"Jhade","suffix":""},{"id":634393344,"identity":"ee683f04-4202-42a2-a95a-e879df1c87c5","order_by":6,"name":"Shradha Devi Dwivedi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABDElEQVRIiWNgGAWjYDACCSDmbWBgbGAGMngMbORAggcekKAlzRisJYEoLSAOD8PhRDADnxb52b0PP7zdYSe74TiP4YM3BYfT54cdfgi0xU5OtwG7FoM7x40l555JNt5wmMfYcI5Beu7G22kGQC3JxmYHcGiRSGOQ5m1jTtx2mC1NmsfAOnfj7ASQlgOJ23BokZ+Rxvybt60epCX9N48Bc7rh7PQPeLUw3EhjA9pyGKiF+Rgzj4Fzgrx0Dn5bDIBaLOe2HTfef5j5sOQcgzTDDdI5BQcSDHD7BeSwG2/bqmVn9h9s/PDmj428/Oz0zR8+VNjJ4dKCxV6wSgNilYPtbSBF9SgYBaNgFIwEAAAYMWPyLV2WmwAAAABJRU5ErkJggg==","orcid":"","institution":"University Institute of Pharmacy, Pandit Ravishankar Shukla University","correspondingAuthor":true,"prefix":"","firstName":"Shradha","middleName":"Devi","lastName":"Dwivedi","suffix":""}],"badges":[],"createdAt":"2026-04-27 09:23:40","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9539312/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9539312/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":108804943,"identity":"8c506c59-58cf-42bf-a7e5-0a2b81d9be43","added_by":"auto","created_at":"2026-05-08 15:24:19","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":610008,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9539312/v1_covered_98664537-fbba-4c9b-95c8-c1a576d81d4a.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"A High Performance Thin Layer Chromatography Method by Design of Experiment approach for quantification of Ascorbic acid in Extracts of Nycthanthus Arbortristis (Parijat) Leaves","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":"Method development, Quantification, Validation, Herbal formulations","lastPublishedDoi":"10.21203/rs.3.rs-9539312/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9539312/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe present study was executed for method development, validation and to quantify Ascorbic acid in Parijat leaves (In- house formulation) and compared with marketed herbal formulations of parijat leaves by high-performance thin-layer chromatography (HPTLC). Isolation of bioactive compounds was carried out using the mobile phase, Methanol: Chloroform : Ammonia (5: 4: 1) and detected at wavelength 273 nm. This method was validated in terms of Accuracy, Precision, Linearity, Limit of detection (LOD) and Limit of quantification (LOQ). The calibration range was found to be 0.5\u0026ndash; 4.5 ng/spot for the active compound. Ascorbic acid was separated with an Rf value 0.55. The method was validated for linearity (0.5\u0026ndash;4.5 ng/spot), accuracy (97\u0026ndash;99%), and RSD of precision (0.13\u0026ndash;1.20%). The LOD and LOQ were found to be 0.0363 ng/spot and 0.1099 ng/spot respectively. The quantification of ascorbic acid in various formulations was in the range of (357.1 \u0026micro;g to 283.8 \u0026micro;g). The design of experiment software was applied by taking three independent variables such as the volume of solvent (methanol), solvent system, and chamber saturation time were investigated for analytical quality by design. This developed HPTLC method is simple, precise, robust, specific, rapid, and cost effective. It could be used for quality control study and quantification of ascorbic acid in the plant and different herbal formulations containing the plant species.\u003c/p\u003e","manuscriptTitle":"A High Performance Thin Layer Chromatography Method by Design of Experiment approach for quantification of Ascorbic acid in Extracts of Nycthanthus Arbortristis (Parijat) Leaves","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-05-06 11:50:51","doi":"10.21203/rs.3.rs-9539312/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":"86cbbeec-5a69-445c-9b3e-2cf5ce8b39dd","owner":[],"postedDate":"May 6th, 2026","published":true,"recentEditorialEvents":[{"type":"editorInvitedReview","content":"","date":"2026-05-11T10:16:07+00:00","index":8,"fulltext":""},{"type":"reviewerAgreed","content":"262896068377183190657375404431455640635","date":"2026-05-04T21:24:44+00:00","index":7,"fulltext":""},{"type":"reviewerAgreed","content":"323784041191150221033492187132304030929","date":"2026-04-30T12:22:28+00:00","index":6,"fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-05-06T11:50:51+00:00","versionOfRecord":[],"versionCreatedAt":"2026-05-06 11:50:51","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9539312","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9539312","identity":"rs-9539312","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}