Automatic Cleaning in Acoustic Ejection Mass Spectrometry: Enhancing the System Robustness for Large-Scale High-Throughput Analysis of Complex Samples

<!DOCTYPE html> <html lang="en" class="pb-page" data-request-id="fa143141-cb4d-4f92-9d36-0b95ba8bed7a" ><head data-pb-dropzone="head"><meta name="pbContext" content=";requestedJournal:journal:slast;website:website:slast-site;ctype:string:Journal Content;journal:journal:slast;pageGroup:string:Article View;wgroup:string:Migrated Websites;page:string:Show Full Text;article:article:pii\:S2472630324001092;subPage:string:Full Text;product:product:elsevier\:product\:ha;csubtype:string:Ahead of Print"/>  <meta name="robots" content="noarchive,nocache" /> <meta name="viewport" content="width=device-width,initial-scale=1,maximum-scale=1, user-scalable=1"/> <meta property="og:title" content="Automatic Cleaning in Acoustic Ejection Mass Spectrometry: Enhancing the System Robustness for Large-Scale High-Throughput Analysis of Complex Samples" /> <meta property="og:type" content="Article" /> <meta property="og:url" content="https://slas-technology.org/article/S2472-6303(24)00109-2/abstract" /> <meta property="og:site_name" content="SLAS Technology" /> <meta property="og:image" content="https://slas-technology.org/cms/asset/ae378187-95d9-4f5b-8b92-0ef413f026b4/gr2.jpg" /> <meta property="og:description" content="The rapid evolution of high-throughput mass spectrometry (HT-MS) technologies has positioned MS as a pivotal analytical tool across diverse disciplines. Its significance is particularly pronounced in high-throughput drug discovery and development, where MS plays a critical role throughout various phases. Acoustic ejection mass spectrometry (AEMS) is a recent addition to the HT-MS landscape, showcasing a balanced performance high analytical throughput and high data quality. Particularly, AEMS's in-line dilution feature allows the direct analysis of large-scale, complex reaction solutions without the need for sample cleanup, making it a popular choice for large-scale high-throughput screenings." /> <meta name="twitter:title" content="Automatic Cleaning in Acoustic Ejection Mass Spectrometry: Enhancing the System Robustness for Large-Scale High-Throughput Analysis of Complex Samples" /> <meta name="twitter:image" content="https://slas-technology.org/cms/asset/ae378187-95d9-4f5b-8b92-0ef413f026b4/gr2.jpg" /> <meta name="twitter:description" content="The rapid evolution of high-throughput mass spectrometry (HT-MS) technologies has positioned MS as a pivotal analytical tool across diverse disciplines. Its significance is particularly pronounced in high-throughput drug discovery and development, where MS plays a critical role throughout various phases. Acoustic ejection mass spectrometry (AEMS) is a recent addition to the HT-MS landscape, showcasing a balanced performance high analytical throughput and high data quality. Particularly, AEMS's in-line dilution feature allows the direct analysis of large-scale, complex reaction solutions without the need for sample cleanup, making it a popular choice for large-scale high-throughput screenings." /> <meta name="twitter:card" content="summary_large_image"> <link rel="canonical" href="https://slas-technology.org/article/S2472-6303(24)00109-2/fulltext"> <title>Automatic Cleaning in Acoustic Ejection Mass Spectrometry: Enhancing the System Robustness for Large-Scale High-Throughput Analysis of Complex Samples - SLAS Technology</title><meta name="citation_journal_title" content="SLAS Technology"/> <meta name="citation_journal_abbrev" content="SLAS Technology"/> <meta name="citation_publisher" content="Elsevier"/> <meta name="citation_title" content="Automatic Cleaning in Acoustic Ejection Mass Spectrometry: Enhancing the System Robustness for Large-Scale High-Throughput Analysis of Complex Samples"/> <meta name="citation_online_date" content="2024/11/22"/> <meta name="citation_volume" content="0"/> <meta name="citation_issue" content="0"/> <meta name="citation_pmid" content="39581265"/> <meta name="citation_abstract_html_url" content="http://slas-technology.org/article/S2472630324001092/abstract"/> <meta name="citation_fulltext_html_url" content="http://slas-technology.org/article/S2472630324001092/fulltext"/> <meta name="citation_pdf_url" content="http://slas-technology.org/article/S2472630324001092/pdf"/> <meta name="citation_issn" content="2472-6303"/> <meta name="citation_issn" content="2472-6311"/> <meta name="citation_language" content="English"/> <meta name="citation_doi" content="10.1016/j.slast.2024.100227"/> <meta name="citation_pmid" content="39581265"/> <meta name="citation_author" content="Heguang Ji"></meta><meta name="citation_author_institution" content="SCIEX, Singapore, 739256, Singapore"></meta><meta name="citation_author" content="Xuejiao Yin"></meta><meta name="citation_author_institution" content="SCIEX, Singapore, 739256, Singapore"></meta><meta name="citation_author" content="Wan Ee Ang"></meta><meta name="citation_author_institution" content="SCIEX, Singapore, 739256, Singapore"></meta><meta name="citation_author" content="Abdullah Bin Rawshan"></meta><meta name="citation_author_institution" content="SCIEX, Singapore, 739256, Singapore"></meta><meta name="citation_author" content="Susan Gay"></meta><meta name="citation_author_institution" content="SCIEX, Singapore, 739256, Singapore"></meta><meta name="citation_author" content="Jing Ma"></meta><meta name="citation_author_institution" content="SCIEX, Singapore, 739256, Singapore"></meta><meta name="citation_author" content="Chiu Cheong Aw"></meta><meta name="citation_author_institution" content="SCIEX, Singapore, 739256, Singapore"></meta><meta name="citation_author" content="Chang Liu"></meta><meta name="citation_author_email" content="Chang.Liu@sciex.com"></meta><meta name="citation_author_institution" content="SCIEX, Concord, Ontario L4K 4V8, Canada"></meta><meta name="citation_keywords" content="high-throughput analysis; mass spectrometry; system robustness; open-port interface; acoustic ejection mass spectrometry; high-throughput screening"></meta><meta name="citation_reference" content="citation_title=Mass spectrometry innovations in drug discovery and development; citation_author=D.I. 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Wagner; citation_publication_date=2023; citation_publisher=SLAS, San Diego, CA, San Diego, CA; "></meta><meta name="citation_reference" content="citation_title=Densities and Viscosities of Methanol-Water Mixtures; citation_author=S. Mikhail; citation_author=W. Kimel; citation_journal_title=Journal of Chemical and Engineering Data; citation_volume=6; citation_number=4; citation_firstpage=533; citation_lastpage=537; citation_publication_date=1961; "></meta><meta name="citation_abstract" content="<h2>Abstract</h2><p>The rapid evolution of high-throughput mass spectrometry (HT-MS) technologies has positioned MS as a pivotal analytical tool across diverse disciplines. Its significance is particularly pronounced in high-throughput drug discovery and development, where MS plays a critical role throughout various phases. Acoustic ejection mass spectrometry (AEMS) is a recent addition to the HT-MS landscape, showcasing a balanced performance high analytical throughput and high data quality. Particularly, AEMS's in-line dilution feature allows the direct analysis of large-scale, complex reaction solutions without the need for sample cleanup, making it a popular choice for large-scale high-throughput screenings. However, the substantial volume of complex matrix introduces concerns about system robustness, specifically regarding the potential clogging of the sample transfer line. This study addresses this challenge by introducing an integrated automatic washing feature to the AEMS system. This enhancement significantly improves system robustness without imposing any additional demands on assay execution time. 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You will then receive an email that contains a secure link for resetting your password</p><div tabindex="-1" class="message error"></div><form action="/action/requestResetPassword" method="post" class="request-reset-password-form"><input type="hidden" name="platform"/><input type="hidden" name="requestResetPassword" value="true"/><input type="hidden" name="codeSite" value="slast-site"/><div class="input-group"><div class="label"><label for="reset-password-email">Email*</label></div><input id="reset-password-email" type="text" name="email" value="" size="15" class="email"/></div><div class="submit"><input type="submit" name="submit" value="Submit" disabled="disabled" class="button primary submit"/></div></form></div><div class="success-template hidden"><p>If the address matches a valid account an email will be sent to __email__ with instructions for resetting your password</p></div></div><div class="request-reset-cancel"><a href="#" aria-label="Close reset password popup" title="Close reset password popup" class="cancel">Cancel</a></div></div></div> <div class="ux3-ads"> <div class="pb-ad">    <div id="gpt-Variables" data-adfile="JBS"> <input type="hidden" id="gptSite" value="slast"> <input type="hidden" id="gptLoggedIn" value="no"> <input type="hidden" id="gptAccessType" value="ae:ANON_GUEST|ae:ANON_GUEST"> </div> <div id="gpt-Interscroller" class="gpt-InScrollBox" style="display:none;"> <div class="gpt-InScrollLabel">ADVERTISEMENT</div> <div class="gpt-InScrollContainer"> <div class="gpt-InScrollWrapper"> <div class="gpt-InScrollInner"> <div class="gpt-adinterscroller" data-adsize="interscroller"></div> </div> </div> </div> <div class="gpt-InScrollLabel">SCROLL TO CONTINUE WITH CONTENT</div> </div> <div id="gpt-DebugDiv" class="gpt-DebugDiv" style="display:none;"> <div style="width: 100%; display: table;"> <div style="display: table-row"> <div style="width: 200px; display: table-cell;"> <button class="gpt-DebugBtn" onclick="gpt_ShowConsole()">Open GPT Console</button><br /> <button id="gpt_GrapeShotDebugBtn" class="gpt-DebugBtn" onclick="gpt_GrapeShotDebug();">Open Oracle Keywords</button><br /> <br /> <button class="gpt-DebugBtn" onclick="gpt_LoadDebugValues()">Refresh Values</button> </div> <div style="display: table-cell;"> <table class="gpt-DebugTable"> <thead> <tr> <th>Property</th> <th>Value</th> </tr> </thead> <tbody> <tr><td class="gpt-DebugTitle">Status</td><td><div class="gptStatus_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Version</td><td><div class="gptVersion_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Ad File</td><td><div class="gptAdFile_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Disable Ads Flag</td><td><div class="gptDisableAds_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Environment</td><td><div class="gptTestENV_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Moat Init</td><td><div class="gptMoatInit_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Moat Ready</td><td><div class="gptMoatReady_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Contextual Ready</td><td><div class="gptContextualReady_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Contextual URL</td><td><div class="gptContextualURL_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Contextual Initial Segments</td><td><div class="gptContextualInit_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Contextual Used Segments</td><td><div class="gptContextualResult_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">AdUnit</td><td><div class="gptSite_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">SubAdUnit</td><td><div class="gptPage_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Custom Targeting</td><td><div class="gptCustomTargeting_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Ad Events</td><td><div class="gptEvent_Debug"></div></td></tr> <tr><td class="gpt-DebugTitle">Invalid Ad Sizes</td><td><div class="gptInvalidAdSize_Debug"></div></td></tr> </tbody> </table> </div> </div> </div> </div> <style type="text/css"> .gpt-DebugBtn { display: inline-block; text-align: center; text-decoration: none; margin: 2px 0; border: solid 1px #122b40; border-radius: 0.4em; padding: 0.5em 1em; color: #ffffff; background-color: #204d74; font-weight: normal; font-size: 12px; } .gpt-DebugDiv { margin: 5px; } .gpt-DebugTitle { font-weight: bold; } .gpt-DebugTable { border: 1px solid #dddddd; border-collapse: collapse; margin: 5px; font-size: 0.9em; font-family: sans-serif; min-width: 400px; margin-bottom: 25px; } .gpt-DebugTable td:first-child { padding: 5px; } .gpt-DebugTable thead tr th { background-color: #d9edf7; color: #ffffff; text-align: left; padding: 5px; border-bottom: 2px solid #204d74; } .gpt-DebugTable th, .gpt-DebugTable td { padding: 5px; } .gpt-DebugTable tbody tr { border-bottom: 1px solid #dddddd; } .gpt-DebugTable tbody tr:nth-of-type(even) { background-color: #f3f3f3; } .gpt-DebugTable tbody tr:last-of-type { border-bottom: 2px solid #204d74; } .gpt-InScrollLabel { background-color: rgb(0, 0, 0); color: rgb(255, 255, 255); position: relative; max-width: 100vw; left: 50%; right: 50%; margin-left: -50vw; margin-right: -50vw; padding: 0; text-align: center; width: 100vw !important; font-size: 10px; } .gpt-InScrollBox { height: 100vh !important; width: 100vw; cursor: pointer; max-width: 100vw; position: relative; left: 50%; right: 50%; margin-left: -50vw; margin-right: -50vw; margin-bottom: 50px; padding-top: 5px; padding-bottom: 5px; z-index: 10001; background-color: white; } .gpt-InScrollBox .gpt-InScrollContainer { height: 95vh !important; position: relative; } .gpt-InScrollBox .gpt-InScrollWrapper { position: absolute !important; top: 0 !important; left: 0; width: 100%; height: 95vh !important; border: 0 !important; margin: 0 !important; padding: 0 !important; clip: rect(0, auto, auto, 0) !important; -webkit-clip-path: polygon(0px 0px, 100% 0px, 100% 100%, 0px 100%) !important; clip-path: polygon(0px 0px, 100% 0px, 100% 100%, 0px 100%) !important; } .gpt-InScrollBox .gpt-InScrollInner { position: fixed !important; top: 0 !important; left: 0 !important; bottom: 0; margin: auto; width: 100vw; height: 95vh; -webkit-transform: translateZ(0) !important; display: -webkit-box; display: -ms-flexbox; display: flex; -webkit-box-orient: vertical; -webkit-box-direction: normal; -ms-flex-direction: column; flex-direction: column; -webkit-box-align: center; -ms-flex-align: center; align-items: center; -webkit-box-pack: center; -ms-flex-pack: center; justify-content: center; text-align: center; } </style> <script class="optanon-category-4" type="text/plain"> //-------------------------------------------------------------------------- var gpt_Version = '04072023'; var gpt_AdFile = $('#gpt-Variables').data('adfile'); var gpt_MaxMOATInitAttempts = 11; var gpt_MaxMOATReadyAttempts = 11; var gpt_MaxContextualReadyAttempts = 21; var gpt_MOATInitAttempts = 0; var gpt_MOATReadyAttempts = 0; var gpt_ContextualReadyAttempts = 0; var gpt_IsProd = true; var gpt_StartTime; var gs_channels = 'DEFAULT'; var gpt_ContextualReady = false; var gpt_ContextualURL = 'Not enabled outside of Production'; var gpt_DisableAdsFlagFound = false; var gptEvent_Debug = ''; var gptCustomTargeting_Debug = ''; var gptSite_Debug = ""; var gptPage_Debug = ""; var gptContextualEnabled = false; var gptContextualResult_Debug = ''; var gptContextualInit_Debug = ''; var gptContextualURL_Debug = ''; var gptContextualReady_Debug = ''; var gptMoatReady_Debug = ''; var gptMoatInit_Debug = ''; var gptTestENV_Debug = ''; var gptDisableAds_Debug = ''; var gptStatus_Debug = 'GPT Still Loading - Refresh Values'; var gptInvalidAdSize_Debug = ''; //-------------------------------------------------------------------------- $(document).ready(function () { //Check to see if we are on a development env gpt_CheckEnvironment(); if (!gpt_DisableAdsFlagFound) { gpt_StartTime = new Date(); if (gpt_AdFile !== "PU") { //Once the dom has loaded, begin the process to init the ads. gpt_InitScripts(); gpt_CheckForMOATInit(); } } }); </script> <script type="text/javascript"> function gpt_CheckEnvironment() { if ($('#gptDisableAds').length > 0) { gptDisableAds_Debug = 'Flag Detected - Ads Disabled'; gpt_DisableAdsFlagFound = true; } else { gptDisableAds_Debug = 'Ads Enabled'; gpt_DisableAdsFlagFound = false; } switch (gpt_AdFile) { case 'PU': gptContextualEnabled = true; if ($("#Environment").val() !== "Prod") { gpt_IsProd = false; gptTestENV_Debug = 'Test Environment Detected'; } else { gptTestENV_Debug = 'Production Environment Detected'; } break; case 'JBS': case 'LANCET': gptContextualEnabled = true; if ($(".gpt-ad").length > 0) { //Other ads exist, allow the Prestitial $(document.body).append("<div class='gpt-ad' data-adsize='prestitial'></div>"); } if (window.location.hostname.toLowerCase().indexOf('.literatumonline.') >= 0) { gpt_IsProd = false; gptTestENV_Debug = 'Test Environment Detected'; } else { gptTestENV_Debug = 'Production Environment Detected'; } break; default: //Cell gptContextualEnabled = false; if (window.location.hostname.toLowerCase().indexOf('.literatumonline.') >= 0) { gpt_IsProd = false; gptTestENV_Debug = 'Test Environment Detected'; } else { gptTestENV_Debug = 'Production Environment Detected'; } break; } } //-------------------------------------------------------------------------- function gpt_GetNonCMEArticle() { var url; $('.articleCitation').each(function (i, obj) { if ($(obj).find('.toc__item__detials').length > 0) { var itemDetails = $(obj).first().find('.toc__item__detials'); var cmeFound = 0; if (cmeFound == 0) { cmeFound = $(itemDetails).find("img[title~='CME']").length; } if (cmeFound == 0) { cmeFound = $(itemDetails).find("img[title~='(CME)']").length; } if (cmeFound == 0) { cmeFound = $(itemDetails).find("img[title~='Education']").length; } if (cmeFound == 0) { cmeFound = $(itemDetails).find("img[title~='education']").length; } if (cmeFound == 0) { var itemLink = $(itemDetails).first().find('.toc__item__title'); var linkCount = $(itemLink).first().find("a:contains('CME')").length; if (linkCount == 0) { url = $(itemLink).first().find('a').attr('href'); return false; //Found a URL, break the loop } } } }); return url; } //-------------------------------------------------------------------------- function gpt_GetElapsedTime() { var endTime = new Date(); var timeDiff = endTime - gpt_StartTime; //in ms // strip the ms timeDiff /= 1000; // get seconds return seconds = Math.round(timeDiff) + " seconds"; } //-------------------------------------------------------------------------- function gpt_CheckSSC() { //Check for ScreenShot Client var gpt_SSC = gpt_GetQuerystring('gpt_ssc', ''); if (gpt_SSC == '1') { //it's the ScreenShotClient, disable MOAT IVT Detection $('#gpt-Variables').append('<input type="hidden" id="gptm_data">'); $('#gptm_data').val(0); gpt_Target("m_data"); } } //-------------------------------------------------------------------------- function gpt_LocalSS(adTypeId) { window.scrollTo(0, 0); gpt_Sleep(1000); gpt_ScrollToAd(adTypeId); gpt_PreTearsheet(adTypeId); } //-------------------------------------------------------------------------- function gpt_InitScripts() { if (gpt_AdFile == "PU") { if (gpt_IsProd) { gpt_ContextualURL = gpt_CheckForNULL($('#canonicalurl').val()); if (gpt_ContextualURL === null) { gpt_ContextualURL = window.location.href.split('?')[0].split('#')[0]; } else { gpt_ContextualURL = window.location.origin + gpt_ContextualURL; } var useSSL = 'https:' == document.location.protocol; var gs = document.createElement('script'); gs.async = true; gs.type = 'text/javascript'; gs.src = (useSSL ? 'https:' : 'http:') + '//elsevier.gscontxt.net/multizone/channels.cgi?url=' + encodeURIComponent(gpt_ContextualURL); gs.onload = function () { gpt_ContextualReady = true; }; document.head.appendChild(gs); } } else { //JBS-Cell-Lancet var useSSL = 'https:' == document.location.protocol; var gads = document.createElement('script'); gads.async = true; gads.type = 'text/javascript'; gads.src = (useSSL ? 'https:' : 'http:') + '//securepubads.g.doubleclick.net/tag/js/gpt.js'; document.head.appendChild(gads); if (gpt_IsProd && gptContextualEnabled) { gpt_ContextualURL = window.location.href.split('?')[0].split('#')[0]; var gs = document.createElement('script'); gs.async = true; gs.type = 'text/javascript'; gs.src = (useSSL ? 'https:' : 'http:') + '//elsevier.gscontxt.net/multizone/channels.cgi?url=' + encodeURIComponent(gpt_ContextualURL); gs.onload = function () { gpt_ContextualReady = true; }; document.head.appendChild(gs); } var elsevierMoatHeader = document.createElement('link'); elsevierMoatHeader.rel = 'preload'; elsevierMoatHeader.as = 'script'; elsevierMoatHeader.src = (useSSL ? 'https:' : 'http:') + '//z.moatads.com/elsevierheader150204004183/moatheader.js'; document.head.appendChild(elsevierMoatHeader); var elsevierMoatAds = document.createElement('link'); elsevierMoatAds.rel = 'preconnect'; elsevierMoatAds.src = (useSSL ? 'https:' : 'http:') + '//mb.moatads.com'; document.head.appendChild(elsevierMoatAds); var moatHeader = document.createElement('script'); moatHeader.async = true; gads.type = 'text/javascript'; moatHeader.src = (useSSL ? 'https:' : 'http:') + '//z.moatads.com/elsevierheader150204004183/moatheader.js'; document.head.appendChild(moatHeader); } } //-------------------------------------------------------------------------- async function gpt_CheckForMOATInit() { //Check every 1/10 of a second until MOAT is Loaded or 1 second timeout if (gpt_MOATInitAttempts < gpt_MaxMOATInitAttempts) { gpt_MOATInitAttempts += 1; if (typeof window.moatPrebidApi === "object") { //MOAT LOADED gptMoatInit_Debug = '0.0' + gpt_MOATInitAttempts + ' seconds'; gpt_CheckForMOATReady(); } else { setTimeout(() => { gpt_CheckForMOATInit(); }, 100); } } else { //TIMEOUT: MOAT Not LOADED gptMoatInit_Debug = 'Timeout Expired'; var googletag = googletag || {}; googletag.cmd = googletag.cmd || []; gpt_CheckForContextualReady(); } } //-------------------------------------------------------------------------- function gpt_CheckForMOATReady() { //Check every 1/10 of a second until MOAT is Ready or 1 second timeout if (gpt_MOATReadyAttempts < gpt_MaxMOATReadyAttempts) { gpt_MOATReadyAttempts += 1; if (window.moatPrebidApi.safetyDataAvailable()) { //MOAT Ready gptMoatReady_Debug = '0.0' + gpt_MOATReadyAttempts + " seconds"; gpt_CheckForContextualReady(); } else { setTimeout(() => { gpt_CheckForMOATReady(); }, 100); } } else { //TIMEOUT: MOAT Not Ready gptMoatReady_Debug = 'Timeout Expired'; gpt_CheckForContextualReady(); } } //-------------------------------------------------------------------------- function gpt_CheckForContextualReady() { //Check every 1/10 of a second until Contextual is Ready or 2 second timeout if (gpt_IsProd && gptContextualEnabled) { //Enabled if (gpt_ContextualReadyAttempts < gpt_MaxContextualReadyAttempts) { gpt_ContextualReadyAttempts += 1; if (gpt_ContextualReady) { //Contextual Ready gptContextualURL_Debug = gpt_ContextualURL; gptContextualReady_Debug = '0.0' + gpt_ContextualReadyAttempts + " seconds"; gptContextualInit_Debug = gs_channels; gpt_InitAds(); } else { setTimeout(() => { gpt_CheckForContextualReady(); }, 100); } } else { //LOG TIMEOUT error //REMOVE FOR NOW (SHOULD LOG TO A DIFF LOCATION) //TIMEOUT: Contextual Not Ready $('#gpt_GrapeShotDebugBtn').remove(); gptContextualURL_Debug = gpt_ContextualURL; gptContextualReady_Debug = 'Timeout Expired'; gptContextualInit_Debug = ""; gpt_InitAds(); } } else { //Not Enabled $('#gpt_GrapeShotDebugBtn').remove(); switch (gpt_AdFile) { case 'PU': case 'LANCET': case 'JBS': gptContextualURL_Debug = 'Not enabled outside of Production'; gptContextualReady_Debug = 'Not enabled outside of Production'; gptContextualInit_Debug = 'Not enabled outside of Production'; break; case "CELLJOURNAL": case "CELLBUCKET": gptContextualReady_Debug = 'Not enabled on Cell'; gptContextualInit_Debug = 'Not enabled on Cell'; gptContextualURL_Debug = 'Not enabled on Cell'; break; } gpt_InitAds(); } } //-------------------------------------------------------------------------- function GPT_GetPageByURL() { var host = window.location.host.toLocaleLowerCase() var localpath = window.location.pathname.toLocaleLowerCase(); ///Check for Cell.com if (host.indexOf('cell.com') >= 0) { return null; } ///Check for Cell.com (marlin-stag) if (host.indexOf('www-cell-com') >= 0) { return null; } ///Check for Lancet if (host.indexOf('thelancet') >= 0) { return null; } if (localpath.indexOf('/article/') >= 0) { return 'article.fulltext'; } if (localpath.indexOf('/action/dosearch') >= 0) { return 'search.searchResults'; } if (localpath.indexOf('/content/') >= 0) { return 'periodical.editorial'; } return 'periodical.home'; } //-------------------------------------------------------------------------- function gpt_CheckForNULL(x, valueifnull) { if (typeof valueifnull == "undefined") { valueifnull = null; } if (typeof x == 'undefined') { return valueifnull; } if (x == null) { return valueifnull; } if (x == "null") { return valueifnull; } if ($.trim(x) == '') { return valueifnull; } return x; } //-------------------------------------------------------------------------- function gpt_GetAdditionalTargeting() { //adds page level targeting of custom variables //the id (minus gpt) and the value will be used as a key-value pair //eg: <input type="hidden" id="gptissuepii" value="x" class="gpt-Target"> //eg: target issuepii=x $(".gpt-Target").each(function (index) { var gptTargetId = $(this).attr('id'); gptTargetId = gptTargetId.replace('gpt', '').toLowerCase(); if (gptTargetId !== '') { gpt_Target(gptTargetId); } }) } //-------------------------------------------------------------------------- function gpt_GetQuerystring(key, default_) { //function to read querystring variables if (default_ === null) default_ = ''; key = key.replace(/[\[]/, "\\\[").replace(/[\]]/, "\\\]"); var regex = new RegExp("[\\?&]" + key + "=([^&#]*)"); var qs = regex.exec(window.location.href); if (qs === null) return default_; else return qs[1]; } //-------------------------------------------------------------------------- function gpt_GetDFPTestId() { //Get any request for a test id from querystring and set custom targeting parameter var gptDFPTestId = gpt_GetQuerystring('dfptestid', ''); if (gptDFPTestId !== '') { $('#gpt-Variables').append('<input type="hidden" id="gptDFPTestId">'); $('#gptDFPTestId').val(gptDFPTestId); gpt_Target("DFPTestId"); } } //-------------------------------------------------------------------------- function gpt_Target(targetName) { //Set page level target if value is found if ($('#gpt' + targetName).length > 0) { var targetValue = $('#gpt' + targetName).val(); if (targetValue !== '') { targetValue = targetValue.replace(/[^-a-z0-9]/ig, ''); //Only Allow a-z, 0-9 googletag.pubads().setTargeting(targetName.toLowerCase(), targetValue.toLowerCase()); gptCustomTargeting_Debug = gptCustomTargeting_Debug + targetName.toLowerCase() + ' = ' + targetValue.toLowerCase() + '<br>'; } } } //-------------------------------------------------------------------------- function gpt_TargetAccessType() { var currentAccessType = $('#gptAccessType').val(); if (typeof currentAccessType == "undefined") { currentAccessType = ""; } if (currentAccessType.indexOf('ANON_IP') >= 0) { googletag.pubads().setTargeting("loggedin", "yes"); } else { gpt_Target("LoggedIn"); } } //-------------------------------------------------------------------------- async function gpt_PreTearsheet(adTypeId) { if (typeof adTypeId === "undefined") { adTypeId = 0; } //Remove Elsevier Cookie Policy Notice if ($("#onetrust-consent-sdk").length > 0) { $("#onetrust-consent-sdk").remove(); } //Mobile Nav $('#skip').remove(); $('#skipNavigationLink').remove(); //JBS Modals if ($('.leadinModal').length > 0) { $('.leadinModal').remove(); } if ($('.leadinModal').length > 0) { $('.leadinModal').remove(); } if ($('.usabilla__overlay').length > 0) { $('.usabilla__overlay').remove(); } //Pendo if ($('._pendo-step-container').length > 0) { $('._pendo-step-container').remove(); } if ($('._pendo-guide-tt_').length > 0) { $('._pendo-guide-tt_').remove(); } if ($('._pendo-image').length > 0) { $('._pendo-image').remove(); } if ($('._pendo-badge-image').length > 0) { $('._pendo-badge-image').remove(); } switch (gpt_AdFile) { case "CELLJOURNAL": case "CELLBUCKET": //Mobile Bug Fix on Cell if (parseInt(adTypeId) == 5) { $('.widget-horizontalAd').css('position', 'static'); $('.body-horizontalAd').css('display', 'block'); } break; case "JBS": case "LANCET": objLeaderboard = ".container-leaderboard"; offset = 53; switch (parseInt(adTypeId)) { case 3: //MPU case 4: //SkyScraper //BUGFIX: Show Sidebar ad for JBS //skyscraper & mpu if ($('.widget-verticalAd').length > 0) { $('.widget-verticalAd').css('display', 'block'); } $('#backgroundLayer').css('display', 'none'); //Hide any Prestitial break; case 6: //Prestitial $('#backgroundLayer').css('display', 'block'); break; default: $('#backgroundLayer').css('display', 'none'); //Hide any Prestitial break; } break; } } //-------------------------------------------------------------------------- function gpt_Sleep(ms) { return new Promise(resolve => setTimeout(resolve, ms)); } //-------------------------------------------------------------------------- async function gpt_ScrollToAd(adTypeId) { if (typeof adTypeId === "undefined") { adTypeId = 0; } var adId = ""; var offset = 0; var objLeaderboard = ""; var gpt_ScrollToAd = true; switch (gpt_AdFile) { case "PU": //PracticeUpdate var hideOtherAds = true; $('#modalPrestitial').modal('hide'); $('.jumbotron-welcome').hide(); objLeaderboard = ".container-leaderboard"; offset = 53; switch (parseInt(adTypeId)) { case 2: //LeaderBoard, no Scroll Needed gpt_ScrollToAd = false; adId = "adSlot1"; break; case 3: //MPU adId = "adSlot2"; break; case 4: //SkyScraper adId = "adSlot3"; $('.most-read').hide(); break; case 5: //Mobile, no Scroll Needed gpt_ScrollToAd = false; adId = "adSlot1"; break; case 6: //Interstitial gpt_ScrollToAd = false; adId = "adSlot15"; await gpt_Sleep(1000); $(objLeaderboard).hide(); $('#modalPrestitial').unbind(); $('#modalPrestitial').modal(); break; case 7: //Native Ad break; case 10: //Master Companion hideOtherAds = false; gpt_ScrollToAd = false; break; default: //Unknown Ad Type gpt_ScrollToAd = false; adId = "adSlot1"; break; } if (hideOtherAds) { //Hide all other ads $('.ad-container').each(function (i, obj) { if (this.id !== adId) { var tempParent = this.id; tempParent = tempParent.replace("adSlot", "adParent"); $("#" + tempParent).addClass('hide'); } }); } break; case "JBS": objLeaderboard = ".js__adToHide"; offset = 80; switch (parseInt(adTypeId)) { case 2: //Leaderboard gpt_ScrollToAd = false; $(".widget-verticalAd").hide(); break; case 5: //Mobile //Scroll to Top gpt_ScrollToAd = false; break; case 3: //MPU case 4: //SkyScraper $(objLeaderboard).hide(); adId = $('.gpt-ad-defined[data-adsize="sidebar"').attr('id'); break; case 10: //MasterCompanion gpt_ScrollToAd = false; break; case 13: //Phone - Portrait case 14: //Phone - Landscape $(objLeaderboard).hide(); offset = 60; adId = 'gpt-Interscroller'; gpt_ScrollToAd = true; break; default: //Unknown Ad Type gpt_ScrollToAd = false; break; } break; case "CELLJOURNAL": case "CELLBUCKET": objLeaderboard = ".js__adToHide"; offset = 260; switch (parseInt(adTypeId)) { case 2: //Leaderboard gpt_ScrollToAd = false; $('.body-verticalAd').hide(); break; case 5: //Mobile //Scroll to Top gpt_ScrollToAd = false; break; case 3: //MPU case 4: //SkyScraper $(objLeaderboard).hide(); adId = $('.gpt-ad-defined[data-adsize="boombox"').attr('id'); break; case 10: //MasterCompanion gpt_ScrollToAd = false; break; default: //Unknown Ad Type gpt_ScrollToAd = false; break; } //Hide the second ad if found var tCount = 0; $(".widget.literatumAd.none.GPT-ad-placeholder.widget-horizontalAd.widget-compact-all").each(function (index) { tCount += 1; if (tCount !== 1) { $(this).hide(); } }); break; case "LANCET": objLeaderboard = ".js__adToHide"; offset = 80; switch (parseInt(adTypeId)) { case 2: //Leaderboard gpt_ScrollToAd = false; $('.body-verticalAd').hide(); break; case 5: //Mobile //Scroll to Top gpt_ScrollToAd = false; break; case 3: //MPU case 4: //SkyScraper $(objLeaderboard).hide(); adId = $('.gpt-ad-defined[data-adsize="skyscraper"').attr('id'); break; case 10: //MasterCompanion gpt_ScrollToAd = false; break; default: //Unknown Ad Type gpt_ScrollToAd = false; break; } break; } //Scroll to Top var gpt_container = $("html,body"); gpt_container.animate({ scrollTop: 0, scrollLeft: 0 }); if (gpt_ScrollToAd) { //If Adexists, scroll to it var gpt_scrollTo = $('#' + adId); if (gpt_scrollTo.length > 0) { //Scroll to the Ad var gpt_topPosition = gpt_scrollTo.offset().top - gpt_container.offset().top + gpt_container.scrollTop(); if (gpt_topPosition > 0) { gpt_container.animate({ scrollTop: gpt_topPosition, scrollLeft: 0 }); if (gpt_topPosition - offset > 0 && offset > 0) { gpt_container.animate({ scrollTop: gpt_topPosition - offset, scrollLeft: 0 }); } } } } } //-------------------------------------------------------------------------- async function gpt_CheckForCookiePolicy() { var returnVal = false; if ($('#onetrust-accept-btn-handler').length > 0) { $('#onetrust-accept-btn-handler').click(); returnVal = true; } return returnVal; } //-------------------------------------------------------------------------- function gpt_InitAds() { var gpt_ResponsiveHomepage = false; switch (gpt_AdFile) { case "JBS": case "CELLJOURNAL": case "CELLBUCKET": //Responsive Check if ($('#hdrHomeBrand').length > 0) { gpt_ResponsiveHomepage = true; } break; } //InterScroller Check if (gpt_AdFile === "JBS") { var gpt_InterScrollerEnabled = false; var gpt_InterScrollerInsertObj; if ($("#articleHeader").length > 0) { //It's an article Page if ($('.article__sections > section').length > 0) { gpt_InterScrollerEnabled = true; gpt_InterScrollerInsertObj = $(".article__sections > section"); } } else { if ($('.toc__section').length > 0) { //It's a TOC Page gpt_InterScrollerEnabled = true; gpt_InterScrollerInsertObj = $(".toc__section").first(); } } //Add the Interscroller if (gpt_InterScrollerEnabled) { gpt_InterScrollerInsertObj.first().append($("#gpt-Interscroller").clone()); $("#gpt-Interscroller").remove(); $(".gpt-adinterscroller").addClass("gpt-ad"); } } googletag.cmd.push(function () { var gptBoomBoxSizeMapping; var gptFluidMapping; var gptHomePageSidebarMapping; var gptCellHomePageSidebarMapping; var gptInterscrollerMapping; var gptLeaderboardSizeMapping; var gptMobileSizeMapping; var gptPrestitialMapping; var gptRectangleShortSizeMapping; var gptRectangleSizeMapping; var gptSideBarSizeMapping; var gptTallSizeMapping; var gptWideSizeMapping; var gptWideRectangleSizeMapping; //*********************************************** //Start Define Size mapping based on client Viewport //*********************************************** if (gpt_AdFile === 'PU') { gptTallSizeMapping = googletag.sizeMapping(). addSize([992, 0], [[300, 600], [160, 600], [120, 600]]). addSize([0, 0], []). build(); gptRectangleSizeMapping = googletag.sizeMapping(). addSize([992, 0], [[300, 250]]). addSize([0, 0], []). build(); gptRectangleShortSizeMapping = googletag.sizeMapping(). addSize([992, 0], [[300, 250], [320, 50], [300, 50]]). addSize([0, 0], [[300, 250], [320, 50], [300, 50]]). build(); gptWideSizeMapping = googletag.sizeMapping(). addSize([992, 0], [[728, 90]]). addSize([760, 0], [[728, 90], [320, 50], [300, 50]]). addSize([0, 0], [[320, 50], [300, 50]]). build(); gptWideRectangleSizeMapping = googletag.sizeMapping(). addSize([992, 0], []). addSize([760, 0], [[320, 50], [300, 50], [300, 250]]). addSize([0, 0], [[320, 50], [300, 50], [300, 250]]). build(); gptMobileSizeMapping = googletag.sizeMapping(). addSize([0, 0], [[320, 50], [300, 50]]). build(); gptFluidMapping = googletag.sizeMapping(). addSize([0, 0], ['fluid']). build(); gptPrestitialMapping = googletag.sizeMapping(). addSize([760, 0], [[1, 1]]). build(); } else { //JBS-CELL-LANCET gptLeaderboardSizeMapping = googletag.sizeMapping(). addSize([992, 0], [[728, 90]]). // Desktop addSize([768, 0], [[728, 90]]). // Tablet Landscape addSize([0, 0], [[320, 50], [300, 50]]). // Phone & Tablet Portrait build(); gptHomePageSidebarMapping = googletag.sizeMapping(). addSize([2090, 795], [[300, 600], [300, 250], [160, 600], [120, 600]]). addSize([1910, 795], [[160, 600], [120, 600]]). addSize([0, 0], []). build(); gptCellHomePageSidebarMapping = googletag.sizeMapping(). addSize([2090, 795], [[300, 600], [300, 250], [336, 280], [160, 600], [120, 600]]). addSize([1910, 795], [[160, 600], [120, 600]]). addSize([0, 0], []). build(); gptSideBarSizeMapping = googletag.sizeMapping(). addSize([1140, 795], [[300, 600], [160, 600], [120, 600], [300, 250]]). // SkyScraper - Desktop (Require min 630 vertical) addSize([1140, 445], [[300, 250]]). // BoomBox Fall Back - Desktop (Require min 280 vertical) addSize([0, 0], []). // Phone build(); gptBoomBoxSizeMapping = googletag.sizeMapping(). addSize([768, 0], [[300, 250], [336, 280]]). // Desktop & Tablet Landscape addSize([0, 0], []). // Phone build(); gptFluidMapping = googletag.sizeMapping(). addSize([0, 0], ['fluid']). build(); gptPrestitialMapping = googletag.sizeMapping(). addSize([600, 700], [[1, 1]]). // Desktop & Tablet Landscape addSize([0, 0], []). // Phone build(); gptInterscrollerMapping = googletag.sizeMapping(). addSize([1140, 445], []). addSize([1024, 768], [[1024, 768]]). addSize([768, 1024], [[768, 1024]]). addSize([480, 320], [[480, 320]]). addSize([320, 480], [[320, 480]]). build(); } //*********************************************** //End Define Size mapping based on client Viewport //*********************************************** //*********************************************** //Start detection for ad slots on page //*********************************************** var gptMasterPage = gpt_CheckForNULL($('#gptPage').val()); var gptMasterSite; if (gpt_AdFile === 'PU') { gptMasterSite = $('#gptSite').val(); } else { //JBS-CELL-LANCET gptMasterSite = "dev"; if (gpt_IsProd) { gptMasterSite = gpt_CheckForNULL($('#gptSite').val()); //If there is no master site, check for a defaultsite if (gptMasterSite == null) { gptMasterSite = gpt_CheckForNULL($('#gptDefaultSite').val()); } } else { //Dev ENV //If the page has no gptSite Value, assume no value which will disable ads if (gpt_CheckForNULL($('#gptSite').val()) === null) { gptMasterSite = null; } } //If there is no master Page, check for a Page using the URL if (gptMasterPage === null) { gptMasterPage = GPT_GetPageByURL(); } } $(".gpt-ad").each(function (index) { var gptPage = ''; var adSizeMapping = ''; var gptPOS; var gptTC; var gptNative; var adId; var gptSite; var tempSlot = null; if (gpt_IsProd) { //Check if the ad slot has it's own site defined if (typeof $(this).data('site') === 'undefined') { gptSite = gptMasterSite; } else { gptSite = gpt_CheckForNULL($(this).data('site')); } } else { gptSite = gptMasterSite; } if (gpt_AdFile === 'PU') { adId = $(this).attr('id'); } else { //JBS-CELL-LANCET adId = 'gpt-ad-' + index + 1; } if (gpt_CheckForNULL(gptSite) !== null) { gptSite_Debug = gptSite; gptPOS = parseInt($(this).data('pos')) || -1; if (gptPOS === -1) { gptPOS = index + 1; } //Check if the ad slot has it's own page defined gptPage = $(this).data('page'); if (gpt_CheckForNULL(gptPage) == null) { //Use the Master Page gptPage = gptMasterPage; } //If the page is not null, preceed it with a slash if (gptPage !== null) { gptPage = '/' + gptPage; } else { gptPage = ''; } gptPage_Debug = gptPage; gptTC = $(this).data('tc'); if (typeof gptTC === "undefined") { gptTC = ''; } gptNative = $(this).data('native'); if (typeof gptNative === "undefined") { gptNative = ''; } //Determine sizemapping based on data-adsize if (gpt_AdFile === 'PU') { switch ($(this).data('adsize')) { case 'tall': adSizeMapping = gptTallSizeMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[120, 600], [160, 600], [300, 600]], adId); break; case 'rectangle': adSizeMapping = gptRectangleSizeMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[300, 250]], adId); break; case 'rectangleshort': adSizeMapping = gptRectangleShortSizeMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[300, 250], [320, 50], [300, 50]], adId); break; case 'wide': adSizeMapping = gptWideSizeMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[728, 90], [320, 50], [300, 50]], adId); break; case 'widerectangle': adSizeMapping = gptWideRectangleSizeMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[320, 50], [300, 50], [300, 250]], adId); break; case 'mobile': adSizeMapping = gptMobileSizeMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[320, 50], [300, 50]], adId); break; case 'fluid': adSizeMapping = gptFluidMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, 'fluid', adId); break; case 'prestitial': adSizeMapping = gptPrestitialMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[1, 1]], adId); break; } } else { //JBS-CELL-LANCET switch ($(this).data('adsize')) { case 'cell-leaderboard': case 'leaderboard': adSizeMapping = gptLeaderboardSizeMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[728, 90], [320, 50], [300, 50]], adId); break; case 'bottom': if (gpt_ResponsiveHomepage) { adSizeMapping = gptHomePageSidebarMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[120, 600], [160, 600], [300, 600], [300, 250]], adId); } else { adSizeMapping = gptBoomBoxSizeMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[300, 250]], adId); } break; case 'boombox': if (gpt_AdFile === "CELLJOURNAL" || gpt_AdFile == "CELLBUCKET") { if (gpt_ResponsiveHomepage) { adSizeMapping = gptCellHomePageSidebarMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[120, 600], [160, 600], [300, 600], [300, 250], [336, 280]], adId); } else { adSizeMapping = gptBoomBoxSizeMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[300, 250], [336, 280]], adId); } } else { adSizeMapping = gptBoomBoxSizeMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[300, 250]], adId); } break; case 'interscroller': adSizeMapping = gptInterscrollerMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[320, 480], [480, 320], [768, 1024], [1024, 768]], adId); break; case 'sidebar': case 'skyscraper': adSizeMapping = gptSideBarSizeMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, [[120, 600], [160, 600], [300, 600], [300, 250]], adId); break; case 'native': case 'fluid': adSizeMapping = gptFluidMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, 'fluid', adId); break; case 'prestitial': adSizeMapping = gptPrestitialMapping; tempSlot = googletag.defineSlot('/6053/els.' + gptSite + gptPage, 'fluid', adId); break; } } } //Ignore any invalid adsizes that could be on the page if (adSizeMapping !== '') { $(this).addClass('gpt-ad-defined'); if (gpt_AdFile !== 'PU') { $(this).attr('id', adId); } tempSlot.defineSizeMapping(adSizeMapping).addService(googletag.pubads()).setTargeting("pos", gptPOS); if (gptTC !== '') { tempSlot.setTargeting("tc", gptTC); } if (gptNative !== '') { tempSlot.setTargeting("native", gptNative); } gptEvent_Debug = gptEvent_Debug + $(this).data('adsize') + ' Requested<br>'; } else { gptInvalidAdSize_Debug = gptInvalidAdSize_Debug + "Invalid Size Found, Id: " + $(this).prop('id') + ', adSize: ' + $(this).data('adsize') + '<br>'; $(this).remove(); } }); if (gpt_CheckForNULL(gptInvalidAdSize_Debug) === null) { gptInvalidAdSize_Debug = "None Detected"; } //*********************************************** //End detection for ad slots on page //*********************************************** //*********************************************** //Start page level targeting //*********************************************** if (gpt_AdFile !== 'PU') { //Logged in or IP Access gpt_TargetAccessType(); //Check for Screenshot Client gpt_CheckSSC(); } //Check for DFP Test gpt_GetDFPTestId(); //Check for Additional Targeting gpt_GetAdditionalTargeting(); if (gpt_IsProd && gptContextualEnabled) { var gpt_ContextualRetry = false; //Check for Contextual Targeting if (gs_channels !== "DEFAULT") { var gpt_ContextualCategories = Array.from(gs_channels); for (var i = 0; i < gpt_ContextualCategories.length; i++) { if (gpt_ContextualCategories[i] == 'gx_retry') { gpt_ContextualRetry = true; } if (!gpt_ContextualCategories[i].startsWith("custom_")) { //Remove any tag that is not custom gpt_ContextualCategories.splice(i, 1); i--; } else { //Strip the 'custom_' from the tag gpt_ContextualCategories[i] = gpt_ContextualCategories[i].replace("custom_", "") } } if (gpt_ContextualCategories.length > 0) { googletag.pubads().setTargeting("gs_cat", gpt_ContextualCategories); } else { gpt_ContextualCategories = 'none'; } if (gpt_ContextualRetry) { gpt_ContextualCategories = "*Indexing*"; } gptContextualResult_Debug = gpt_ContextualCategories; } else { gptContextualResult_Debug = 'No segments found'; } } else { //Not Prod gptContextualResult_Debug = 'Not enabled'; var gpt_DevSite = gpt_CheckForNULL($('#gptSite').val(), "dev"); $('#gpt-Variables').append('<input type="hidden" id="gptEnvironment" value="' + gpt_DevSite + '">'); gpt_Target("Environment"); } //*********************************************** //End page level targeting //*********************************************** //Enable google dfp services if (parseInt($('#gptLazyLoading').val()) === 1) { googletag.pubads().enableLazyLoad({ fetchMarginPercent: 200, renderMarginPercent: 0, mobileScaling: 2.0 }); } else { googletag.pubads().enableSingleRequest(); } //Add Listener for RenderEnded googletag.pubads().addEventListener('slotRenderEnded', function (event) { if (!event.isEmpty) { if (typeof LocalAdLoaded === "function") { LocalAdLoaded(event); } if (gpt_ResponsiveHomepage) { if (typeof positionAd === "function") { positionAd(); } } if (event.size[0] == "480" || event.size[1] == "480") { $('.gpt-InScrollBox').css('display', 'block'); gptEvent_Debug = gptEvent_Debug + 'Interscroller ' + event.size + ' rendered: ' + gpt_GetElapsedTime() + '<br>'; } else { gptEvent_Debug = gptEvent_Debug + event.size + ' rendered: ' + gpt_GetElapsedTime() + '<br>'; } } }); if (gpt_AdFile !== 'PU') { googletag.pubads().collapseEmptyDivs(); } googletag.enableServices(); //Populate Defined Ad Slots $(".gpt-ad-defined").each(function (index) { var adId = $(this).attr('id'); googletag.cmd.push(function () { googletag.display(adId); }); }); gptStatus_Debug = 'GPT Complete'; }); } //-------------------------------------------------------------------------- function gpt_DisplayDebug() { //Clone the template and append to Body $(document.body).append($('#gpt-DebugDiv').clone()); $('#gpt-DebugDiv').remove(); //Populate the Values gpt_LoadDebugValues(); //Display and scroll to debug section $('.gpt-DebugDiv').css('display', 'block'); window.scrollTo(0, document.body.scrollHeight); } //-------------------------------------------------------------------------- function gpt_LoadDebugValues() { $('.gpt-DebugBtn').blur(); $('.gptVersion_Debug').text(gpt_Version); $('.gptAdFile_Debug').html(gpt_AdFile); $('.gptEvent_Debug').html(gptEvent_Debug); $('.gptCustomTargeting_Debug').html(gptCustomTargeting_Debug); $('.gptSite_Debug').text(gptSite_Debug); $('.gptContextualResult_Debug').text(gptContextualResult_Debug); $('.gptContextualInit_Debug').text(gptContextualInit_Debug); $('.gptContextualURL_Debug').text(gptContextualURL_Debug); $('.gptContextualReady_Debug').text(gptContextualReady_Debug); $('.gptMoatReady_Debug').text(gptMoatReady_Debug); $('.gptMoatInit_Debug').text(gptMoatInit_Debug); $('.gptTestENV_Debug').text(gptTestENV_Debug); $('.gptDisableAds_Debug').text(gptDisableAds_Debug); $('.gptStatus_Debug').text(gptStatus_Debug); $('.gptPage_Debug').text(gptPage_Debug); $('.gptInvalidAdSize_Debug').html(gptInvalidAdSize_Debug); } //-------------------------------------------------------------------------- function gpt_GrapeShotDebug() { $('.gpt-DebugBtn').blur(); window.open("http://admin-elsevier.grapeshot.co.uk/custom/bookmarklet.cgi?blocking=1&vn=2&url=" + encodeURIComponent(document.location), "GRAPESHOT_POPUP", "width=650,height=600,scrollbars=yes,screenX=40,screenY=50,menubar=no,toolbar=no,resizable=yes", "Custom").focus(); } //-------------------------------------------------------------------------- function gpt_ShowConsole() { $('.gpt-DebugBtn').blur(); googletag.openConsole(); } //-------------------------------------------------------------------------- </script> </div> </div> <div data-widget-id="53023812-5eb3-476f-8922-6f6f95be0a5c" data-widget-name="ux3-layout-widget"><header class="header base fixed"> <div class="widget literatumAd none adplaceholder adplaceholder--header js__adToHide widget-horizontalAd" id="ad532c9c-4e0f-4b0d-8b27-9bd0ddbdb190"> <div class="wrapped " > <div class="widget-body body body-horizontalAd "><div class="horizontalAdContainer"> <div class="horizontalAdLabelContainer"> <div class="horizontalAdLabel"> <img src="/templates/jsp/_style2/_marlin/images/bg_adHoriz.gif" alt="Advertisement"/> </div> </div> <div class="horizontalAd"> <div class="pb-ad"> <div class="gpt-ad" data-adsize="leaderboard"></div> </div> </div> </div></div> </div> </div> <div data-widget-def="ux3-layout-widget" data-widget-id="3bd430f7-bd17-4546-89a2-5b0ca3b18cdf" class="header__wrapper clearfix" data-db-parent-of="quickSearch"> <div 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url("/templates/jsp/css/jquery-ui-1.10.2/base/images/ui-icons_454545_256x240.png"); margin-right: 5px; margin-top: -4px; } .csas-institution-banner .banner-close { float: right; } </style> <script> function resetBanner(doi) { $.get('/action/getAccount?doi=' + doi, function(data) { if (data) { $(".csas-institution-banner p .accountName").html(data); $(".csasInstitutionBanner").fadeIn(); $(".csas-institution-banner").fadeIn(); } }); } var flag = 0; $(document).ready(function() { if(!$(".csas-institution-banner").is(":visible")){ $(".csasInstitutionBanner").hide(); } $(".banner-close").click(function() { $.get("/action/dismissBanner", function(data) { $(".csasInstitutionBanner").fadeOut(function () { if(typeof UX !== 'undefined') { UX.pageBody.init(); } }); }); }); $(".articleContent .tabs li[role='tab'].showFullText").click(function() { flag = 1; }); $(document).on("reloadTabsContent", function() { var isSelected = $(".articleContent .tabs 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vocab="http://schema.org/" typeof="ScholarlyArticle" data-pii="S2472-6303(24)00109-2" lang="en" dir="ltr"><div data-core-wrapper="header"><header data-extent="frontmatter"><div class="core-container"><div data-article-access="full" data-article-access-type="open" class="meta-panel"><span class="meta-panel__left"><span class="meta-panel__type"><span>Short Communication</span></span></span><span class="meta-panel__middle"><span class="meta-panel__volumeIssue"><a href="/inpress"><span>Articles in Press</span></a></span><span class="meta-panel__number"><span>100227</span></span><span class="meta-panel__onlineDate">November 22, 2024</span></span><span class="meta-panel__right"><span class="meta-panel__access meta-panel__access--open"><span>Open access</span></span></span></div><h1 property="name">Automatic Cleaning in Acoustic Ejection Mass Spectrometry: Enhancing the System Robustness for Large-Scale High-Throughput Analysis of Complex Samples</h1><div class="contributors"><span class="authors"><span role="list"><span property="author" typeof="Person" role="listitem"><a href="#au0001"><span property="givenName">Heguang</span> <span property="familyName">Ji</span></a><sup class="xref"><a href="#aff0001" property="affiliation" typeof="Organization" role="doc-noteref">†</a>,<a href="#fn1" role="doc-noteref">§</a></sup></span> ∙ <span property="author" typeof="Person" role="listitem"><a href="#au0002"><span property="givenName">Xuejiao</span> <span property="familyName">Yin</span></a><sup class="xref"><a href="#aff0001" property="affiliation" typeof="Organization" role="doc-noteref">†</a>,<a href="#fn1" role="doc-noteref">§</a></sup></span> ∙ <span property="author" typeof="Person" role="listitem"><a href="#au0003"><span property="givenName">Wan Ee</span> <span property="familyName">Ang</span></a><sup class="xref"><a href="#aff0001" property="affiliation" typeof="Organization" role="doc-noteref">†</a></sup></span><span data-displayed-on="all"> ∙ … </span><span data-hidden-on="all"> ∙ <span property="author" typeof="Person" role="listitem"><a href="#au0004"><span property="givenName">Abdullah Bin</span> <span property="familyName">Rawshan</span></a><sup class="xref"><a href="#aff0001" property="affiliation" typeof="Organization" role="doc-noteref">†</a></sup></span></span><span data-hidden-on="all"> ∙ <span property="author" typeof="Person" role="listitem"><a href="#au0005"><span property="givenName">Susan</span> <span property="familyName">Gay</span></a><sup class="xref"><a href="#aff0001" property="affiliation" typeof="Organization" role="doc-noteref">†</a></sup></span></span> ∙ <span property="author" typeof="Person" role="listitem"><a href="#au0006"><span property="givenName">Jing</span> <span property="familyName">Ma</span></a><sup class="xref"><a href="#aff0001" property="affiliation" typeof="Organization" role="doc-noteref">†</a></sup></span> ∙ <span property="author" typeof="Person" role="listitem"><a href="#au0007"><span property="givenName">Chiu Cheong</span> <span property="familyName">Aw</span></a><sup class="xref"><a href="#aff0001" property="affiliation" typeof="Organization" role="doc-noteref">†</a></sup></span> ∙ <span property="author" typeof="Person" role="listitem" class="corresponding-author"><a href="#au0008"><span property="givenName">Chang</span> <span property="familyName">Liu</span></a><sup class="xref"><a href="#aff0002" property="affiliation" typeof="Organization" role="doc-noteref">‡</a></sup> <a id="ead0001" href="mailto:Chang.Liu@sciex.com" property="email" title="Send email to Chang Liu">Chang.Liu@sciex.com</a></span><span data-displayed-on="all"> … <span><span data-action="reveal">Show more</span></span> </span><span data-displayed-on="none" data-on-display="all" aria-hidden="true"> <span data-action="hide">Show less</span></span></span></span></div><div class="core-self-citation"><div id="core-affiliations-notes"><div class="affiliations"><div property="affiliation" typeof="Organization"><span class="label">†</span><span id="aff0001" property="name">SCIEX, Singapore, 739256, Singapore</span></div><div property="affiliation" typeof="Organization"><span class="label">‡</span><span id="aff0002" property="name">SCIEX, Concord, Ontario L4K 4V8, Canada</span></div></div><div class="core-authors-notes"><div role="doc-footnote" data-has="label"><div class="label">§</div><div id="fn1"><div id="notep0001" role="paragraph">These authors contributed equally to this work.</div></div></div></div></div><div id="core-content-info"><div class="history"><div>Publication History:</div><div><span>Received September 12, 2024</span>; <span>Revised November 12, 2024</span>; <span>Accepted November 22, 2024</span>; <span>Published online November 22, 2024</span></div></div><div class="core-links"><span class="doi">DOI: <a href="https://doi.org/10.1016/j.slast.2024.100227" property="sameAs" target="_blank">10.1016/j.slast.2024.100227</a></span><span class="science-direct">Also available on <a href="https://www.sciencedirect.com/science/article/pii/S2472630324001092" target="_blank">ScienceDirect</a></span></div><div role="paragraph">Copyright: © 2024 The Authors. 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aria-hidden="true" class="icon-arrow_l"></i><span>Previous article</span></a><a href="/article/S2472-6303(24)00107-9/fulltext" role="menuitem" aria-label="Next article" aria-disabled="false" class="content-navigation__next"><span>Next article</span><i aria-hidden="true" class="icon-arrow_r"></i></a></div></div></div></div></li></ul></div><div><div class="core-content-inner"><div class="core-linked-content"></div><div id="abstracts" data-extent="frontmatter"><div class="core-container"><section id="author-abstract" property="abstract" typeof="Text" role="doc-abstract"><h2 property="name">Abstract</h2><div id="spara008" role="paragraph">The rapid evolution of high-throughput mass spectrometry (HT-MS) technologies has positioned MS as a pivotal analytical tool across diverse disciplines. Its significance is particularly pronounced in high-throughput drug discovery and development, where MS plays a critical role throughout various phases. Acoustic ejection mass spectrometry (AEMS) is a recent addition to the HT-MS landscape, showcasing a balanced performance high analytical throughput and high data quality. Particularly, AEMS's in-line dilution feature allows the direct analysis of large-scale, complex reaction solutions without the need for sample cleanup, making it a popular choice for large-scale high-throughput screenings. However, the substantial volume of complex matrix introduces concerns about system robustness, specifically regarding the potential clogging of the sample transfer line. This study addresses this challenge by introducing an integrated automatic washing feature to the AEMS system. This enhancement significantly improves system robustness without imposing any additional demands on assay execution time. Demonstrating an extended electrode lifetime, the cleaning approach proves effective in maintaining system performance over prolonged periods, showcasing its potential for continuous large-sample-scale high-throughput analysis applications.</div></section></div></div><div class="core-linked-content"></div><section id="keywords" property="keywords"><h2>Keywords</h2><ol><li><a href="/action/doSearch?AllField="high-throughput+analysis"&journalCode=slast" alt="Search on this keyword">high-throughput analysis</a></li><li><a href="/action/doSearch?AllField="mass+spectrometry"&journalCode=slast" alt="Search on this keyword">mass spectrometry</a></li><li><a href="/action/doSearch?AllField="system+robustness"&journalCode=slast" alt="Search on this keyword">system robustness</a></li><li><a href="/action/doSearch?AllField="open-port+interface"&journalCode=slast" alt="Search on this keyword">open-port interface</a></li><li><a href="/action/doSearch?AllField="acoustic+ejection+mass+spectrometry"&journalCode=slast" alt="Search on this keyword">acoustic ejection mass spectrometry</a></li><li><a href="/action/doSearch?AllField="high-throughput+screening"&journalCode=slast" alt="Search on this keyword">high-throughput screening</a></li></ol></section><div class="core-linked-content"></div><div class="core-linked-content"></div><div class="core-linked-content"></div><section id="bodymatter" data-extent="bodymatter" property="articleBody" typeof="Text"><div class="core-container"><section id="sec-1"><h2>1 Introduction</h2><div id="para0002" role="paragraph">Mass spectrometry (MS) has become a cornerstone in drug discovery due to its high specificity, broad dynamic range, and high sensitivity, making it a pivotal analytical platform for delivering high-fidelity, label-free analysis results [<a id="crf0012" href="#bib0001" role="doc-biblioref" data-xml-rid="bib0001">1</a>]. Liquid chromatography (LC) is the predominant method to deliver liquid samples to MS through electrospray ionization (ESI). Despite its success, the analytical throughput is constrained to around 30 seconds per sample [<a id="crf0013" href="#bib0002" role="doc-biblioref" data-xml-rid="bib0002">2</a>], limiting the application of LC-MS in high-throughput settings. This restriction is particularly evident in workflows requiring the analysis of a large sample set, such as high-throughput pharmacology screening [<a id="crf0014" href="#bib0003" role="doc-biblioref" data-xml-rid="bib0003">3</a>,<a id="crf0015" href="#bib0004" role="doc-biblioref" data-xml-rid="bib0004">4</a>], reaction monitoring for high-throughput experimentation [<a id="crf0016" href="#bib0005" role="doc-biblioref" data-xml-rid="bib0005">5</a>,<a id="crf0017" href="#bib0006" role="doc-biblioref" data-xml-rid="bib0006">6</a>], and quality control of compound libraries [<a id="crf0018" href="#bib0007" role="doc-biblioref" data-xml-rid="bib0007">7</a>].</div><div id="para0003" role="paragraph">In a quest to enhance MS analysis throughput, various LC-free non-ESI approaches have been explored, including matrix-assisted laser desorption ionization (MALDI) [<a id="crf0019" href="#bib0008" role="doc-biblioref" data-xml-rid="bib0008 bib0009 bib0010 bib0011 bib0012 bib0013">8–13</a>], laser diode thermal desorption (LDTD) [<a id="crf0021" href="#bib0014" role="doc-biblioref" data-xml-rid="bib0014">14</a>], direct analysis in real time (DART) [<a id="crf0022" href="#bib0015" role="doc-biblioref" data-xml-rid="bib0015">15</a>], desorption electrospray ionization (DESI) [<a id="crf0023" href="#bib0016" role="doc-biblioref" data-xml-rid="bib0016">16</a>,<a id="crf0024" href="#bib0017" role="doc-biblioref" data-xml-rid="bib0017">17</a>], matrix-assisted laser desorption electrospray ionization (MALDESI) [<a id="crf0025" href="#bib0018" role="doc-biblioref" data-xml-rid="bib0018">18</a>,<a id="crf0026" href="#bib0019" role="doc-biblioref" data-xml-rid="bib0019">19</a>], and acoustic mist ionization (AMI) [<a id="crf0027" href="#bib0020" role="doc-biblioref" data-xml-rid="bib0020 bib0021 bib0022">20–22</a>]. Despite their rapid analysis speeds, many of these approaches necessitate sample treatment, introducing additional cost and automation complexity. Moreover, the integration of sampling and ionization as a single step may compromise ionization performance, leading to challenges like matrix effects and limited coverage of the chemical space, restricting the applicability of these technologies in complex sample matrix analysis.</div><div id="para0004" role="paragraph">To tackle these challenges, the acoustic ejection mass spectrometry (AEMS) technology has emerged as a solution for high-throughput analysis [<a id="crf0029" href="#bib0023" role="doc-biblioref" data-xml-rid="bib0023">23</a>,<a id="crf0030" href="#bib0024" role="doc-biblioref" data-xml-rid="bib0024">24</a>]. Utilizing the open-port interface (OPI), AEMS segregates the high-throughput and high-precision sampling facilitated by acoustic droplet ejection (ADE) from the classic electrospray ionization (ESI) process. This innovative approach enables the retention of advantages from both high-throughput MS and LC-MS simultaneously. Since its introduction, it has been utilized in a wide range of high-throughput analysis workflows including drug discovery and development [<a id="crf0031" href="#bib0025" role="doc-biblioref" data-xml-rid="bib0025 bib0026 bib0027 bib0028">25–28</a>], synthetic biology [<a id="crf0033" href="#bib0029" role="doc-biblioref" data-xml-rid="bib0029">29</a>,<a id="crf0034" href="#bib0030" role="doc-biblioref" data-xml-rid="bib0030">30</a>], and the poultry industry [<a id="crf0035" href="#bib0031" role="doc-biblioref" data-xml-rid="bib0031">31</a>]. One of the unique features of the AEMS technology is the in-line matrix dilution. In this process, the nanoliter-sized sample droplet is captured within the carrier solvent being delivered at a flow rate of 300-600 µL/min, the sample undergoes substantial dilution before ionization. This mitigates ionization suppression [<a id="crf0036" href="#bib0032" role="doc-biblioref" data-xml-rid="bib0032">32</a>], enabling the direct analysis of complex matrices without the need for extensive sample cleanup, simplifying sample preparation, and facilitating the analysis of unstable analytes from high-salt-concentration buffers [<a id="crf0037" href="#bib0033" role="doc-biblioref" data-xml-rid="bib0033">33</a>].</div><div id="para0005" role="paragraph">However, the large number of analyses of the complex matrix raises the concerns about system robustness, particularly the risk of OPI transfer line clogging, which potentially disrupts the high-throughput analysis workflow in an unmonitored automation-controlled environment [<a id="crf0038" href="#bib0034" role="doc-biblioref" data-xml-rid="bib0034">34</a>]. In this study, we introduce an integrated automatic cleaning setup within the AEMS system. The washing process is automatically triggered during the plate exchange process without extending the assay execution time. Demonstrating an extended electrode lifetime of at least three times for representative pharmacology screening assays, this novel cleaning approach underscores its efficacy in maintaining system performance over prolonged periods.</div></section><section id="sec-2"><h2>2 Materials and methods</h2><section id="sec-2-1"><h3>2.1 Reagents and materials</h3><div id="para0006" role="paragraph">Dextromethorphan was acquired from Cerilliant (Round Rock, TX), and the triazine standard mixture was obtained from SCIEX (Framingham, MA). HPLC-grade methanol and acetonitrile were sourced from Caledon Laboratory Chemicals (Georgetown, ON, Canada). Acetic acid and ammonium formate were procured from Honeywell Fluka (Phoenix, AZ). Tris (hydroxymethyl) aminomethane, magnesium chloride, potassium chloride, citric acid, bovine serum albumin (BSA), ammonium acetate and Tween20 were obtained from Merck (Darmstadt, Germany). 1,4-Dithiothreitol (DTT) was acquired from Roche (Basel, Switzerland). Deionized water (18 MΩ) was generated in-house using a Millipore (Billerica, MA) Integral 10 water purification system. Echo MS qualified 384-well plates were supplied by Beckman Coulter Life Sciences (San Jose, CA).</div></section><section id="sec-2-2"><h3>2.2 AEMS system</h3><div id="para0007" role="paragraph">The working principles of the AEMS system have been comprehensively described in previous studies [<a id="crf0039" href="#bib0023" role="doc-biblioref" data-xml-rid="bib0023 bib0024">23–24</a>]. This technology employs an OPI to connect the high-speed acoustic droplet ejection with ESI-MS. The sample droplets acoustically ejected from 384- and 1536-well plates are precisely captured and diluted within a continuous carrier solvent flow within the OPI, subsequently delivered to ESI for direct analysis. The noncontact sampling facilitated by ADE and the continuous self-washing feature of OPI effectively eliminate cross-contaminations between samples. The in-line dilution of sample droplets within the carrier solvent in the OPI significantly reduces ionization suppression, enabling the direct analysis of complex samples from incubation buffers without the need for sample cleanup.</div><div id="para0008" role="paragraph">The prototype AEMS systems used in this study consisted of a SCIEX ZenoTOF 7600 or Triple Quad 6500+ mass spectrometer equipped with a standard OptiFlow ESI source and an Echo MS module (SCIEX, Concord, ON, Canada) housing the modified solvent delivering system. Detailed information on system modifications is discussed in the subsequent section. The whole system was operated using a prototype version of SCIEX OS software. Sample plates underwent centrifugation at 1530g for 2 minutes before loading onto the system to remove bubbles from the solution and keep the constant meniscus shape for each sample. The ion spray voltage was set at 5 kV, and the source temperature was maintained at 300°C. GS1 and GS2 pressures were configured at 90 and 70 psi, respectively, while the curtain gas was regulated at 25 psi. Subsequent to the acquisition of MS data from all acoustic ejections, data analysis was executed using the same prototype version of SCIEX OS.</div></section></section><section id="sec-3"><h2>3 Results and discussion</h2><section id="sec-3-1"><h3>3.1 OPI washing requirements</h3><div id="para0009" role="paragraph">Due to its distinctive advantages of balanced high analytical throughput and high data quality, AEMS technology has found extensive use across various workflows, particularly in scenarios involving the analysis of a large number of samples. Large-scale sample analyses, often involving tens to hundreds of microplates per day, are typically conducted in a fully automated environment without human intervention. Ensuring data quality and sample integrity in such high-throughput settings requires a robust system to facilitate uninterrupted screening. In the AEMS system, nanoliter-sized samples are acoustically ejected from microplates, captured in the continuous carrier flow within the OPI. The in-line dilution of sample matrices within the OPI, up to several thousand folds before the ESI process, enables direct analysis of the original reaction incubation buffer. This unique feature significantly simplifies sample preparation and method development, facilitating the analysis of unstable analytes present in complex buffers, such as those retaining protein activity in covalent binders screening workflows [<a id="crf0041" href="#bib0033" role="doc-biblioref" data-xml-rid="bib0033">33</a>]. Moreover, it enables the investigation of reaction kinetics in real-time (<i>in situ</i> kinetics study) during the ongoing reaction [<a id="crf0042" href="#bib0024" role="doc-biblioref" data-xml-rid="bib0024">24</a>].</div><div id="para0010" role="paragraph">In the AEMS system, organic carrier solvents such as methanol and acetonitrile are commonly employed due to their low viscosity, allowing high flow rates and analytical throughput [<a id="crf0043" href="#bib0032" role="doc-biblioref" data-xml-rid="bib0032">32</a>]. However, the low solubility of salts and proteins in these solvents poses a risk of matrix precipitation within the OPI after prolonged accumulation of large-scale sample ejections, despite the ejection volume for each sample being in the nanoliter range. This raises the potential for partial clogging of the transfer tube linking the OPI to the ion source, potentially compromising data quality. The number of samples analyzed before the transfer-line starts showing signs of clogging depends on factors such as sample composition, carrier solvent type and flow rate, and ejection volume for each sample. Studies indicate stable signal performance through 400,000 ejections in a typical high-throughput screening assay with methanol as the carrier solvent [<a id="crf0044" href="#bib0035" role="doc-biblioref" data-xml-rid="bib0035">35</a>]. Assays using acetonitrile, which has a lower salt solubility than methanol, may exhibit the Z’ starting to fall below 0.5 for poor assay robustness after approximately 46,000 ejections [<a id="crf0045" href="#bib0025" role="doc-biblioref" data-xml-rid="bib0025">25</a>].</div><div id="para0011" role="paragraph">As the precipitates are typically aqueous-soluble, using a solution containing water has been reported to redissolve salts and improve electrode lifetime [<a id="crf0046" href="#bib0034" role="doc-biblioref" data-xml-rid="bib0034">34</a>]. The team from Boehringer Ingelheim demonstrated the execution of a 1.5-million-sample high-throughput screening assay with a single electrode by incorporating periodic washing steps [<a id="crf0047" href="#bib0025" role="doc-biblioref" data-xml-rid="bib0025">25</a>]. However, these reported OPI washing processes either require manual disassembly of the electrode and flushing the tubing with a separate pump [<a id="crf0048" href="#bib0025" role="doc-biblioref" data-xml-rid="bib0025">25</a>] or involve bypassing the solvent delivery system in the integrated AEMS system, relying on heavily customized automation software to control the additional solvent pumps and the ADE module of the system [<a id="crf0049" href="#bib0025" role="doc-biblioref" data-xml-rid="bib0025">25</a>]. In this study, we introduce a fully integrated AEMS system with automatic scheduled OPI cleaning to enhance system robustness.</div><div id="para0012" role="paragraph">Several requirements guide the development of this automatic OPI cleaning feature. The washing process should ideally not add any additional assay execution time and be seamlessly integrated during the plate exchange process. In the AEMS system, this plate exchange takes approximately 3 minutes, including plate extension to the unloading position, plate swapping by the automation arm, retraction of the new plate into the system together with deionization, plate classification, and the pre-screening plate survey. The integrated control software receives a signal indicating that plate analysis has finished, triggering the OPI washing process immediately and ensuring completion before the screening of the next plate. In this study, the washing cycle is controlled to finish within this 3-minute plate exchange duration, with flexibility for adjustment based on specific sample buffers or automation-controlled plate swapping speeds. Another requirement is the effectiveness of the OPI cleaning, which typically necessitates the use of a different solvent composition (containing more aqueous component) from the carrier solvent, requiring a separate set of solvent delivering pump systems. Additionally, considering the higher viscosity of water, precautions are taken to avoid potential OPI overflow that might contaminate the coupling fluid of the acoustic ejection module. The system design addressing these requirements will be detailed in the following section.</div></section><section id="sec-3-2"><h3>3.2 System modification and control synchronization</h3><div id="para0013" role="paragraph">In the standard AEMS system, the carrier solvent is drawn from the solvent bottle to the degasser using a gear pump to eliminate potential dissolved gases. The degasser outlet is directly connected to a pump equipped with a flowmeter for precise and accurate solvent delivery. The pump outlet is linked to a diverter valve, controlling the flow either to the OPI for AEMS operations or to a waste container during pump purging. In this study, a second, independently controlled pump system was introduced to deliver the washing solvent. This secondary system includes an additional solvent bottle, degasser, and pump. Calibration of flow rates for water, methanol, and acetonitrile combinations is equipped to ensure precise control over a broad range of washing solvents. Two 6-port rotary valves are employed to achieve different flow conditions (<a id="crf0050" href="#fig0001">Figure 1</a>A). <a id="crf0051" href="#fig0001">Figure 1</a>B (1B/2A) illustrates the assay screening mode, where the carrier solvent is pumped directly to the OPI. <a id="crf0052" href="#fig0001">Figure 1</a>C (1A/2A) represents the standard purging mode, where the carrier solvent is pumped directly into the waste bottle. These modes perform similar functions as the conventional setup. Configuration 1B/2B (<a id="crf0053" href="#fig0001">Figure 1</a>D) allows the washing solvent to flow to the OPI device, while configuration 1A/2B (<a id="crf0054" href="#fig0001">Figure 1</a>E) can be activated to purge the solvent pump and move the remaining liquid within the OPI.</div><div class="figure-wrap"><figure id="fig0001" class="graphic"><a class="icon-full-screen" href="/cms/10.1016/j.slast.2024.100227/asset/7d9f3129-cdca-4c96-a782-b1f9c9e39863/main.assets/gr1_lrg.jpg" target="_blank" title="View full size image in a new tab"><img alt="Figure 1" src="/cms/10.1016/j.slast.2024.100227/asset/f2680091-56a4-4d23-9fff-fe10dac1e9c9/main.assets/gr1.jpg" height="861" width="791" data-viewer-src="/cms/10.1016/j.slast.2024.100227/asset/7d9f3129-cdca-4c96-a782-b1f9c9e39863/main.assets/gr1_lrg.jpg" loading="lazy" /></a><figcaption class="figure__caption__body"><div id="fig0001-title" class="figure__title"><span class="label figure__label">Figure 1</span> <span class="figure__title__text">Fluidic delivery module of the system used in this study. A. The connections between each component. B-E, fluidic flowing pathways with different value positions.</span></div></figcaption></figure></div><div id="para0014" role="paragraph"><a id="crf0055" href="#fig0002">Figure 2</a> illustrates the flowrate profile from the two pumps in a complete washing cycle lasting approximately 3 minutes, aligning with the plate exchange time. Consequently, the OPI washing process seamlessly integrates between plate screenings without extending assay execution time. This process is initiated by the system control software once data acquisition concludes. During the OPI washing process, the washing solvent is aspirated to the ESI ion source using the same Venturi pull as during the plate screening. Due to the higher viscosity of the washing solvent containing water, OPI overflow may occur if the same flow rate is used for the washing solvent as the carrier solvent, leading to contamination of the coupling water in the acoustic ejection system. To address this challenge, specific considerations have been incorporated into the method design.</div><div class="figure-wrap"><figure id="fig0002" class="graphic"><a class="icon-full-screen" href="/cms/10.1016/j.slast.2024.100227/asset/03977ed2-72c9-4e70-b696-e6e23e4e8b15/main.assets/gr2_lrg.jpg" target="_blank" title="View full size image in a new tab"><img alt="Figure 2" src="/cms/10.1016/j.slast.2024.100227/asset/ae378187-95d9-4f5b-8b92-0ef413f026b4/main.assets/gr2.jpg" height="461" width="508" data-viewer-src="/cms/10.1016/j.slast.2024.100227/asset/03977ed2-72c9-4e70-b696-e6e23e4e8b15/main.assets/gr2_lrg.jpg" loading="lazy" /></a><figcaption class="figure__caption__body"><div id="fig0002-title" class="figure__title"><span class="label figure__label">Figure 2</span> <span class="figure__title__text">Flow profiles of an OPI cleaning cycle.</span></div></figcaption></figure></div><div id="para0015" role="paragraph">Initially, a lower flow rate for the washing solvent (phase 2) than the carrier solvent is employed. The chosen washing solvent flow rate is determined based on the viscosity relationship between the carrier flow and the washing solvent. For example, if the operational carrier methanol (0.55 cp) flow rate is approximately 450 µL/min, the washing solvent (50:50 methanol-water) with approximately three times the viscosity (1.54 cp) could be adjusted to a flow rate of about 150 µL/min without causing OPI overflow. Additionally, a liquid removal step is applied before each solvent exchange (phase 1 and 3) to prevent mixing between carrier and washing solvents, as the combination of organic solvent and water may lead to a nonlinear increase in viscosity higher than water [<a id="crf0056" href="#bib0032" role="doc-biblioref" data-xml-rid="bib0032">32</a>,<a id="crf0057" href="#bib0036" role="doc-biblioref" data-xml-rid="bib0036">36</a>]. Furthermore, a recovery duration of lower flow rate of the carrier solvent is applied before returning the flow rate to the normal operational range. Five different carrier solvents and washing solvents were tested as outlined in <a id="crf0058" href="#tbl0001">Table 1</a>, with 20 washing cycles tested for each condition. No OPI overflow was observed during these tests. To ensure no contamination of the coupling water, a sensor detecting overflow dripping was activated to halt the liquid pump upon occurrence, and a rotary tray was positioned above the acoustic transducer assembly during the washing process to collect any overflow (<a id="crf0059" href="#fig0003">Figure 3</a>).</div><div class="figure-wrap"><figure id="fig0003" class="graphic"><a class="icon-full-screen" href="/cms/10.1016/j.slast.2024.100227/asset/53e5e5ab-2882-4740-97e2-df60bff547d2/main.assets/gr3_lrg.jpg" target="_blank" title="View full size image in a new tab"><img alt="Figure 3" src="/cms/10.1016/j.slast.2024.100227/asset/8fea03d2-d6de-443a-9197-24e48d52373d/main.assets/gr3.jpg" height="277" width="678" data-viewer-src="/cms/10.1016/j.slast.2024.100227/asset/53e5e5ab-2882-4740-97e2-df60bff547d2/main.assets/gr3_lrg.jpg" loading="lazy" /></a><figcaption class="figure__caption__body"><div id="fig0003-title" class="figure__title"><span class="label figure__label">Figure 3</span> <span class="figure__title__text">(A) The drip sensor surrounding the OPI in the prototype AEMS system to detect the OPI overflow. (B)The rotary tray placed in between of OPI and the acoustic ejection module during the OPI washing process.</span></div></figcaption></figure></div><div class="figure-wrap"><figure id="tbl0001" class="table" data-figureviewer-ignore="true"><div class="table-wrap"><table data-figureviewer-ignore="true"><thead><tr class="rowsep" data-xml-align="left" data-xml-valign="top"><th id="en0001">Solvent Combination</th><th id="en0002">Carrier Solvent Type</th><th id="en0003">Carrier Solvent Flowrate (µL/min)</th><th id="en0004">Wash Solvent Type</th><th id="en0005">Wash Solvent<br />Flowrate (µL/min)</th></tr></thead><tbody><tr><td id="en0006">A</td><td id="en0007">100% MeOH</td><td id="en0008">450</td><td id="en0009">Water/MeOH 80:20</td><td id="en0010">150</td></tr><tr><td id="en0011">B</td><td id="en0012">ACN/MeOH 50:50 + 2mM NH<sub>4</sub>COOH</td><td id="en0013">610</td><td id="en0014">Water/MeOH 70:30 + 0.1% acetic acid</td><td id="en0015">150</td></tr><tr><td id="en0016">C</td><td id="en0017">100% ACN</td><td id="en0018">500</td><td id="en0019">100% Water</td><td id="en0020">150</td></tr><tr><td id="en0021">D</td><td id="en0022">100% MeOH</td><td id="en0023">390</td><td id="en0024">100% Water</td><td id="en0025">150</td></tr><tr><td id="en0026">E</td><td id="en0027">100% MeOH</td><td id="en0028">450</td><td id="en0029">Water/MeOH 50:50</td><td id="en0030">150</td></tr></tbody></table></div><figcaption><span class="heading">Table 1</span><div id="spara006" role="paragraph">Carrier solvents and washing solvents tested for evaluating the risks of OPI overflow.</div><div class="core-table-tools"><ul><li><a class="core-table-show-action" href="/action/showFullTableHTML?isHtml=true&tableId=tbl0001&pii=S2472-6303%2824%2900109-2" target="_blank">Open table in a new tab</a></li></ul></div></figcaption></figure></div></section><section id="sec-3-3"><h3>3.3 Performance characterizations</h3><div id="para0016" role="paragraph">The OPI washing feature was initially characterized by assessing the AEMS performance immediately after the washing process. <a id="crf0060" href="#tbl0002">Table 2</a> provides a summary of the analytical performance for a complete 384-plate run with a mixture of standards (triazine mixture) before and after washing. Demonstrated reproducibility was observed in peak area, signal duration, signal-to-noise ratio, and mass accuracy. There was no residual washing solvent that could adversely affect the system's performance. The comparison of full scan spectra of the carrier solvent before and after washing (<a id="crf0061" href="#fig0004">Figure 4</a>) confirmed consistent ion patterns, validating the complete removal of the washing solvent.</div><div class="figure-wrap"><figure id="fig0004" class="graphic"><a class="icon-full-screen" href="/cms/10.1016/j.slast.2024.100227/asset/67866806-d937-4691-9471-40f4b5c895c4/main.assets/gr4_lrg.jpg" target="_blank" title="View full size image in a new tab"><img alt="Figure 4" src="/cms/10.1016/j.slast.2024.100227/asset/2afb5799-6263-413d-8a1e-fea3678289d0/main.assets/gr4.jpg" height="211" width="678" data-viewer-src="/cms/10.1016/j.slast.2024.100227/asset/67866806-d937-4691-9471-40f4b5c895c4/main.assets/gr4_lrg.jpg" loading="lazy" /></a><figcaption class="figure__caption__body"><div id="fig0004-title" class="figure__title"><span class="label figure__label">Figure 4</span> <span class="figure__title__text">Comparison of background ions before and after the OPI washing</span></div></figcaption></figure></div><div class="figure-wrap"><figure id="tbl0002" class="table" data-figureviewer-ignore="true"><div class="table-wrap"><table data-figureviewer-ignore="true"><thead><tr data-xml-align="left" data-xml-valign="top"><th id="en0031" colspan="6"><span>Before OPI cleaning</span></th><th id="en0032" colspan="6"><span>After OPI cleaning</span></th></tr><tr class="rowsep" data-xml-align="left" data-xml-valign="top"><th id="en0033" colspan="2"><span>Parameters</span></th><th id="en0034">Prometryn</th><th id="en0035">Ametryn</th><th id="en0036">Atrazine</th><th id="en0037">Prometon</th><th id="en0038" colspan="2"><span>Parameters</span></th><th id="en0039">Prometryn</th><th id="en0040">Ametryn</th><th id="en0041">Atrazine</th><th id="en0042">Prometon</th></tr></thead><tbody><tr><td id="en0043" rowspan="3">Peak area</td><td id="en0044">Average</td><td id="en0045">1.66e4</td><td id="en0046">9.38e3</td><td id="en0047">6.12e3</td><td id="en0048">2.03e4</td><td id="en0049" rowspan="3">Peak area</td><td id="en0050">Average</td><td id="en0051">1.78e4</td><td id="en0052">1.00e4</td><td id="en0053">6.70e3</td><td id="en0054">2.18e4</td></tr><tr><td id="en0056">Std Dev</td><td id="en0057">915.94</td><td id="en0058">581.17</td><td id="en0059">414.41</td><td id="en0060">1080.69</td><td id="en0062">Std dev</td><td id="en0063">1191.03</td><td id="en0064">683.56</td><td id="en0065">475.40</td><td id="en0066">1451.32</td></tr><tr><td id="en0068">CV (%)</td><td id="en0069">5.52</td><td id="en0070">6.20</td><td id="en0071">6.77</td><td id="en0072">5.33</td><td id="en0074">CV (%)</td><td id="en0075">6.70</td><td id="en0076">6.81</td><td id="en0077">7.09</td><td id="en0078">6.65</td></tr><tr><td id="en0079" colspan="2">Peak width (sec)</td><td id="en0080">0.37</td><td id="en0081">0.37</td><td id="en0082">0.37</td><td id="en0083">0.37</td><td id="en0084" colspan="2">Peak width (sec)</td><td id="en0085">0.37</td><td id="en0086">0.37</td><td id="en0087">0.37</td><td id="en0088">0.37</td></tr><tr><td id="en0089" colspan="2">Signal/noise</td><td id="en0090">19.34</td><td id="en0091">19.48</td><td id="en0092">18.763</td><td id="en0093">15.26</td><td id="en0094" colspan="2">Signal/noise</td><td id="en0095">19.51</td><td id="en0096">19.00</td><td id="en0097">18.74</td><td id="en0098">16.03</td></tr><tr><td id="en0099" colspan="2">Mass accuracy (ppm)</td><td id="en0100">1.43</td><td id="en0101">1.46</td><td id="en0102">1.03</td><td id="en0103">1.62</td><td id="en0104" colspan="2">Mass accuracy (ppm)</td><td id="en0105">0.99</td><td id="en0106">1.11</td><td id="en0107">0.72</td><td id="en0108">0.93</td></tr></tbody></table></div><figcaption><span class="heading">Table 2</span><div id="spara007" role="paragraph">Performance comparison before and after the OPI washing.</div><div class="core-table-tools"><ul><li><a class="core-table-show-action" href="/action/showFullTableHTML?isHtml=true&tableId=tbl0002&pii=S2472-6303%2824%2900109-2" target="_blank">Open table in a new tab</a></li></ul></div></figcaption></figure></div><div id="para0017" role="paragraph">The enhancement of AEMS system robustness in prolonging electrode lifetime is exemplified through a representative high-throughput biochemical screening involving salts, proteins, reducing agents, and detergents (buffer details listed in <a id="crf0062" href="#fig0005">Figure 5</a> caption). The carrier solvent, a mixture of methanol and acetonitrile containing salts and acids representing situations of low salt solubility. The sample's ejection volume was 10 nL, as reported in a prior HTS study [<a id="crf0063" href="#bib0025" role="doc-biblioref" data-xml-rid="bib0025">25</a>]. Without OPI washing, analytical performance deteriorated after approximately 100,000 ejections, with signals merging and becoming irreproducible at plate 248. Subsequent plates showed no distinguishable signals due to partial OPI transfer line blockage caused by salt precipitation (<a id="crf0064" href="#fig0005">Figure 5</a>A). In contrast, with OPI cleaning between screening plates, analytical performance remained consistent even after 330,000 ejections (860 plates), significantly improving system uptime without extending assay execution durations (<a id="crf0065" href="#fig0005">Figure 5</a>B). This approach also led to a notable reduction in the frequency of electrode replacements, enhancing the efficiency of uninterrupted screening.</div><div class="figure-wrap"><figure id="fig0005" class="graphic"><a class="icon-full-screen" href="/cms/10.1016/j.slast.2024.100227/asset/1d7172dc-fc3f-40a4-8ea8-b06af852191b/main.assets/gr5_lrg.jpg" target="_blank" title="View full size image in a new tab"><img alt="Figure 5" src="/cms/10.1016/j.slast.2024.100227/asset/93ee58cc-9aba-4b76-9fc2-7c67da7c774a/main.assets/gr5.jpg" height="746" width="791" data-viewer-src="/cms/10.1016/j.slast.2024.100227/asset/1d7172dc-fc3f-40a4-8ea8-b06af852191b/main.assets/gr5_lrg.jpg" loading="lazy" /></a><figcaption class="figure__caption__body"><div id="fig0005-title" class="figure__title"><span class="label figure__label">Figure 5</span> <span class="figure__title__text">The comparison of AEMS system robustness. A. Chronogram of the plate 1, plate 248 and plate 249. There was no OPI cleaning between plates. B. Chronogram of the plate 1, plate 500 and plate 860. There was OPI cleaning between plates.</span></div></figcaption></figure></div></section></section><section id="sec-4"><h2>4 Conclusions</h2><div id="para0018" role="paragraph">This study introduces an integrated automatic cleaning setup in the AEMS system, operating during the plate exchange without extending assay execution time. Demonstrating an extended electrode lifetime with pharmacology high-throughput screening assay conditions, this novel cleaning approach ensures sustained system performance over prolonged periods, mitigating risks associated with unmonitored, automation-controlled workflows. The demonstrated feature highlights the efficacy of the cleaning system in enhancing AEMS reliability, thereby facilitating its broader application in diverse high-throughput analysis workflows.</div></section><section id="sec-5"><h2>CRediT authorship contribution statement</h2><div id="para0018a" role="paragraph"><b>Heguang Ji:</b> Writing – review & editing, Data curation. <b>Xuejiao Yin:</b> Writing – review & editing, Data curation. <b>Wan Ee Ang:</b> Writing – review & editing, Data curation. <b>Abdullah Bin Rawshan:</b> Writing – review & editing, Data curation. <b>Susan Gay:</b> Writing – review & editing, Data curation. <b>Jing Ma:</b> Writing – review & editing, Data curation. <b>Chiu Cheong Aw:</b> Writing – review & editing, Data curation. <b>Chang Liu:</b> Writing – review & editing, Writing – original draft, Investigation, Conceptualization.</div></section><section id="conflict" class="core-conflict"><h2>Declaration of competing interest</h2><div id="para0019" role="paragraph">The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:</div><div id="para0020" role="paragraph">Heguang Ji, Xuejiao, Yin, Wan Ee Ang, Abdullah Bin Rawshan, Susan Gay, Jing Ma, Chiu Cheong Aw, Chang Liu reports a relationship with SCIEX that includes: employment. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.</div></section><section id="acknowledgments" role="doc-acknowledgments"><h2>Acknowledgements</h2><div id="para0021" role="paragraph">The authors would like to acknowledge Bogdan Georgescu, David Cox, and Lyle Burton from SCIEX for their assistance for the data processing software used in this study. We also thank Wilson Shou from Bristol-Myers Squibb, Tim T. Häbe from Boehringer Ingelheim, and Thomas R. 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property="author" typeof="Person"><div class="heading"><span property="givenName">Heguang</span> <span property="familyName">Ji</span></div><div class="content"><div class="core-author-notes"><div class="label">Footnotes</div><div role="doc-footnote" data-has="label"><div class="label">§</div><div id="fn1"><div id="notep0001" role="paragraph">These authors contributed equally to this work.</div></div></div></div><div class="core-author-affiliations"><div class="label">Affiliations</div><div property="affiliation" typeof="Organization"><span property="name">SCIEX, Singapore, 739256, Singapore</span></div></div><div class="core-author-link"><a href="/authored-by/Ji/Heguang" target="_blank">Search for articles by this author</a></div></div></div><div id="au0002" property="author" typeof="Person"><div class="heading"><span property="givenName">Xuejiao</span> <span property="familyName">Yin</span></div><div class="content"><div class="core-author-notes"><div class="label">Footnotes</div><div role="doc-footnote" data-has="label"><div class="label">§</div><div id="fn1"><div id="notep0001" role="paragraph">These authors contributed equally to this work.</div></div></div></div><div class="core-author-affiliations"><div class="label">Affiliations</div><div property="affiliation" typeof="Organization"><span property="name">SCIEX, Singapore, 739256, Singapore</span></div></div><div class="core-author-link"><a href="/authored-by/Yin/Xuejiao" target="_blank">Search for articles by this author</a></div></div></div><div id="au0003" property="author" typeof="Person"><div class="heading"><span property="givenName">Wan Ee</span> <span property="familyName">Ang</span></div><div class="content"><div class="core-author-affiliations"><div class="label">Affiliations</div><div property="affiliation" typeof="Organization"><span property="name">SCIEX, Singapore, 739256, Singapore</span></div></div><div class="core-author-link"><a href="/authored-by/Ang/Wan+Ee" 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Automatic Cleaning in Acoustic Ejection Mass Spectrometry: Enhancing the System Robustness for Large-Scale High-Throughput Analysis of Complex Samples - SLAS Technology