{"id":22,"date":"2026-03-11T13:02:19","date_gmt":"2026-03-11T13:02:19","guid":{"rendered":"https:\/\/site.uvm.edu\/alca\/hplc-dad-fld\/"},"modified":"2026-05-19T15:01:17","modified_gmt":"2026-05-19T15:01:17","slug":"hplc-dad-fld","status":"publish","type":"page","link":"https:\/\/site.uvm.edu\/alca\/instrumentation\/hplc-dad-fld\/","title":{"rendered":"High Performance Liquid Chromatography (HPLC)"},"content":{"rendered":"\n<p class=\"wp-block-paragraph\">The Agilent 1260 Infinity HPLC System provides robust and flexible separation of compounds in solution using a range of detection methods. The system supports multiple detectors, enabling both qualitative and quantitative analysis across diverse sample types.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Available detectors include&nbsp;<strong>refractive index (RID), diode array detection (DAD), and fluorescence detection (FLD)<\/strong>, allowing analysis of compounds with varying optical and chemical properties.<\/p>\n\n\n\n<div class=\"wp-block-columns is-layout-flex wp-container-core-columns-is-layout-adc57f8e wp-block-columns-is-layout-flex\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<div class=\"wp-block-group has-global-padding is-layout-constrained wp-block-group-is-layout-constrained\">\n<h2 class=\"wp-block-heading has-primary-uvmgreen-color has-text-color has-link-color wp-elements-305ccc4f07ef498590e0a42b8cab6a0c\">Detection Methods<\/h2>\n<\/div>\n\n\n\n<div class=\"wp-block-group has-global-padding is-layout-constrained wp-block-group-is-layout-constrained\">\n<div class=\"wp-block-group has-global-padding is-layout-constrained wp-block-group-is-layout-constrained\">\n<h3 class=\"wp-block-heading\">Refractive Index Detector (RID)<\/h3>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Universal detector for compounds lacking UV absorbance<\/li>\n\n\n\n<li>Ideal for:\n<ul class=\"wp-block-list\">\n<li>Sugars<\/li>\n\n\n\n<li>Polymers (gel permeation chromatography (GPC) and size exclusion chromatography (SEC))<\/li>\n\n\n\n<li>Alcohols<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n\n\n\n<h3 class=\"wp-block-heading\">Diode Array Detector (DAD)<\/h3>\n\n\n\n<ul class=\"wp-block-list\">\n<li>UV-Vis<\/li>\n\n\n\n<li>Simultaneously collects data at multiple wavelengths<\/li>\n\n\n\n<li>Provides spectral information<\/li>\n\n\n\n<li>Ideal for:\n<ul class=\"wp-block-list\">\n<li>Compound identification <\/li>\n\n\n\n<li>Quantification <\/li>\n\n\n\n<li>Purity assessment <\/li>\n<\/ul>\n<\/li>\n<\/ul>\n\n\n\n<h3 class=\"wp-block-heading\">Fluorescence Detector (FLD)<\/h3>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Highly sensitive for fluorescent compounds <\/li>\n\n\n\n<li>Lower detection limits than UV<\/li>\n\n\n\n<li>Ideal for:\n<ul class=\"wp-block-list\">\n<li>Biomolecules <\/li>\n\n\n\n<li>Trace-level detection<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<div class=\"wp-block-group has-global-padding is-layout-constrained wp-block-group-is-layout-constrained\">\n<h2 class=\"wp-block-heading has-text-align-center has-primary-uvmgreen-color has-text-color has-link-color wp-elements-73d1b9638430ed8ebc40fea22e4cedde\">Fraction Collection Capability<\/h2>\n\n\n\n<h4 class=\"wp-block-heading\"><strong>Agilent 1260 Infinity II Analytical-Scale Fraction Collector<\/strong><\/h4>\n\n\n\n<p class=\"wp-block-paragraph\">The system is equipped with an analytical-scale fraction collector, enabling&nbsp;<strong>isolation and collection of separated compounds following chromatographic separation<\/strong>.<\/p>\n\n\n\n<h3 class=\"wp-block-heading\">What This Enables<\/h3>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Isolation of purified compounds<\/strong>&nbsp;from complex mixtures<\/li>\n\n\n\n<li><strong>Collection of individual peaks<\/strong>&nbsp;based on retention time or detector signal<\/li>\n\n\n\n<li><strong>Preparation of samples<\/strong>&nbsp;for:\n<ul class=\"wp-block-list\">\n<li>LC-MS (QTOF)<\/li>\n\n\n\n<li>NMR analysis<\/li>\n\n\n\n<li>Further chemical characterization<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n\n\n\n<h3 class=\"wp-block-heading\">Compatible Column Dimensions<\/h3>\n\n\n\n<p class=\"wp-block-paragraph\">The system is compatible with a range of analytical preparative HPLC columns, including:<\/p>\n\n\n\n<h4 class=\"wp-block-heading\">Analytical Columns<\/h4>\n\n\n\n<p class=\"wp-block-paragraph\">Typical dimensions:<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>2.1 mm ID<\/strong><\/li>\n\n\n\n<li><strong>3.0 mm ID<\/strong><\/li>\n\n\n\n<li><strong>4.6 mm ID<\/strong><\/li>\n<\/ul>\n<\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-group has-global-padding is-layout-constrained wp-block-group-is-layout-constrained\">\n<h2 class=\"wp-block-heading has-text-align-center has-primary-uvmgreen-color has-text-color has-link-color wp-elements-bb852d8247b1f0954506804a62f69aa5\">Sample Requirements <\/h2>\n\n\n\n<h4 class=\"wp-block-heading\">Solvent &amp; Buffer Compatibility<\/h4>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Samples should be <strong>fully dissolved in LC-compatible solvents<\/strong>\n<ul class=\"wp-block-list\">\n<li>Aqueous\/organic mixtures (e.g. water, methanol, acetonitrile)<\/li>\n<\/ul>\n<\/li>\n\n\n\n<li>Preferred LC-MS  or HPLC grade solvents<\/li>\n<\/ul>\n\n\n\n<h4 class=\"wp-block-heading\">Sample Cleanliness<\/h4>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Filter samples using <strong>0.2 \u00b5m or 0.45 \u00b5m filters<\/strong> or alternatively centrifuge to remover particles <\/li>\n\n\n\n<li>Particles can \n<ul class=\"wp-block-list\">\n<li>Clog the LC system<\/li>\n\n\n\n<li>Damage the column<\/li>\n\n\n\n<li>Affect reproducibility<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n\n\n\n<h4 class=\"wp-block-heading\">Concentration Guidelines<\/h4>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Typical working range:\n<ul class=\"wp-block-list\">\n<li><strong>\u00b5M to mM<\/strong> for most analyses<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<\/div>\n\n\n\n<h2 class=\"wp-block-heading has-text-align-center has-primary-uvmgreen-color has-text-color has-link-color wp-elements-4c85efac5d18e729f8830f671bdb74b4\">Rates<\/h2>\n\n\n\n<figure class=\"wp-block-table is-style-stripes is-style-stripes--1\"><table class=\"has-fixed-layout\"><thead><tr><th class=\"has-text-align-center\" data-align=\"center\">Service (per hour)<\/th><th class=\"has-text-align-center\" data-align=\"center\">UVM Users<\/th><th class=\"has-text-align-center\" data-align=\"center\">External Users<\/th><\/tr><\/thead><tbody><tr><td class=\"has-text-align-center\" data-align=\"center\">User Instrument Time<\/td><td class=\"has-text-align-center\" data-align=\"center\">$19.94<\/td><td class=\"has-text-align-center\" data-align=\"center\">$29.91<\/td><\/tr><tr><td class=\"has-text-align-center\" data-align=\"center\">Staff Assisted Use<\/td><td class=\"has-text-align-center\" data-align=\"center\">$91.87<\/td><td class=\"has-text-align-center\" data-align=\"center\">$137.80<\/td><\/tr><tr><td class=\"has-text-align-center\" data-align=\"center\">Training Session<\/td><td class=\"has-text-align-center\" data-align=\"center\">$91.87<\/td><td class=\"has-text-align-center\" data-align=\"center\">$137.81<\/td><\/tr><\/tbody><tfoot><tr><td class=\"has-text-align-center\" data-align=\"center\">Prices as of July 1, 2025<\/td><td class=\"has-text-align-center\" data-align=\"center\"><\/td><td class=\"has-text-align-center\" data-align=\"center\"><\/td><\/tr><\/tfoot><\/table><\/figure>\n\n\n\n<div class=\"wp-block-columns is-layout-flex wp-container-core-columns-is-layout-adc57f8e wp-block-columns-is-layout-flex\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<h3 class=\"wp-block-heading has-text-align-center has-primary-uvmgreen-color has-text-color has-link-color wp-elements-fbc6f45e5064b4a755f9026bba77c833\">Training<\/h3>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Interested in running the HPLC independently?<\/strong><br>Schedule training to learn method development, detector selection, and data analysis.<\/p>\n<\/div>\n\n\n\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<h3 class=\"wp-block-heading has-text-align-center has-primary-uvmgreen-color has-text-color has-link-color wp-elements-81cf02e4f1bc8ebb8729caa940f0a538\">Analysis<\/h3>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Need help with separations or quantitation?<\/strong><br>Submit samples for full-service HPLC analysis, including method development and data interpretation.<\/p>\n<\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-buttons is-layout-flex wp-block-buttons-is-layout-flex\">\n<div class=\"wp-block-button has-custom-width wp-block-button__width-50\"><a class=\"wp-block-button__link has-white-color has-primary-uvmgreen-background-color has-text-color has-background has-link-color wp-element-button\" href=\"https:\/\/site.uvm.edu\/alca\/contact\/\">Schedule Training<\/a><\/div>\n\n\n\n<div class=\"wp-block-button has-custom-width wp-block-button__width-50\"><a class=\"wp-block-button__link has-primary-uvmgreen-background-color has-background wp-element-button\" href=\"https:\/\/uvm.ilab.agilent.com\/service_center\/show_external\/6442?name=laboratory-for-chemical-analysis\">Request Analysis on iLabs<\/a><\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>The Agilent 1260 Infinity HPLC System provides robust and flexible separation of compounds in solution using a range of detection methods. The system supports multiple detectors, enabling both qualitative and quantitative analysis across diverse sample types. Available detectors include&nbsp;refractive index (RID), diode array detection (DAD), and fluorescence detection (FLD), allowing analysis of compounds with varying [&hellip;]<\/p>\n","protected":false},"author":10857,"featured_media":179,"parent":20,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-22","page","type-page","status-publish","has-post-thumbnail","hentry"],"_links":{"self":[{"href":"https:\/\/site.uvm.edu\/alca\/wp-json\/wp\/v2\/pages\/22","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/site.uvm.edu\/alca\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/site.uvm.edu\/alca\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/site.uvm.edu\/alca\/wp-json\/wp\/v2\/users\/10857"}],"replies":[{"embeddable":true,"href":"https:\/\/site.uvm.edu\/alca\/wp-json\/wp\/v2\/comments?post=22"}],"version-history":[{"count":11,"href":"https:\/\/site.uvm.edu\/alca\/wp-json\/wp\/v2\/pages\/22\/revisions"}],"predecessor-version":[{"id":459,"href":"https:\/\/site.uvm.edu\/alca\/wp-json\/wp\/v2\/pages\/22\/revisions\/459"}],"up":[{"embeddable":true,"href":"https:\/\/site.uvm.edu\/alca\/wp-json\/wp\/v2\/pages\/20"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/site.uvm.edu\/alca\/wp-json\/wp\/v2\/media\/179"}],"wp:attachment":[{"href":"https:\/\/site.uvm.edu\/alca\/wp-json\/wp\/v2\/media?parent=22"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}