{"id":4979,"date":"2026-09-18T16:25:16","date_gmt":"2026-09-18T14:25:16","guid":{"rendered":"https:\/\/novogin.cc\/?p=4979"},"modified":"2026-09-25T14:26:28","modified_gmt":"2026-09-25T12:26:28","slug":"hplc-en-lc-ms","status":"publish","type":"post","link":"https:\/\/novogin.com\/en\/hplc-en-lc-ms\/","title":{"rendered":"HPLC and LC-MS: what does each method measure?"},"content":{"rendered":"<p class=\"wp-block-paragraph\">Certificates of analysis often mention HPLC and LC-MS. These techniques provide different types of information about the same sample. This guide mainly compares HPLC with UV detection and LC-MS. LC-MS also uses liquid chromatography: the distinction partly lies in the detector and the question the laboratory is investigating.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"hplc-lc-ms-vergeleken\" style=\"margin-top:48px;margin-bottom:16px;font-size:clamp(18.959px, 1.185rem + ((1vw - 3.2px) * 0.863), 30px);line-height:1.25\">HPLC met UV-detectie en LC-MS vergeleken<\/h2>\n\n\n\n<figure class=\"wp-block-table\"><table><thead><tr><th>Vraag<\/th><th>HPLC met UV-de\u00adtec\u00adtie<\/th><th>LC-MS<\/th><\/tr><\/thead><tbody><tr><td>Wat wordt ge\u00adme\u00adten?<\/td><td>UV-res\u00adpons van ge\u00adschei\u00adden com\u00adpo\u00adnen\u00adten tegen de tijd.<\/td><td>Massa-la\u00adding\u00adver\u00adhou\u00addin\u00adgen van ionen, ge\u00adkop\u00adpeld aan de chro\u00adma\u00adto\u00adgra\u00adfi\u00adsche schei\u00adding.<\/td><\/tr><tr><td>Wat helpt dit be\u00adoor\u00adde\u00adlen?<\/td><td>Het piek\u00adpa\u00adtroon en, bin\u00adnen de ge\u00adbruik\u00adte me\u00adtho\u00adde, re\u00adla\u00adtie\u00adve piek\u00adop\u00adper\u00advlak\u00adken.<\/td><td>Of ge\u00adme\u00adten ionen pas\u00adsen bij de ver\u00adwach\u00adte massa en la\u00addings\u00adtoe\u00adstan\u00adden.<\/td><\/tr><tr><td>Wat volgt er niet van\u00adzelf uit?<\/td><td>E\u00e9n piek of een hoog per\u00adcen\u00adta\u00adge be\u00adwijst op zich\u00adzelf geen iden\u00adti\u00adteit of hoe\u00adveel\u00adheid in mil\u00adli\u00adgram.<\/td><td>Een pas\u00adsen\u00adde massa be\u00adwijst op zich\u00adzelf niet de vol\u00adle\u00addi\u00adge ami\u00adno\u00adzuur\u00advolg\u00ador\u00adde.<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n<p class=\"wp-block-paragraph\">Een kwantitatieve uitkomst vraagt bij beide technieken een geschikte meetopzet. Lees het gerapporteerde resultaat, de eenheid en de kalibratie; leid de hoeveelheid niet af uit alleen een grafiek of methodenaam.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"hplc\" style=\"margin-top:48px;margin-bottom:16px;font-size:clamp(18.959px, 1.185rem + ((1vw - 3.2px) * 0.863), 30px);line-height:1.25\">What does HPLC do?<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">HPLC stands for High Performance Liquid Chromatography. A liquid carries the sample through a column. Compounds travel through it differently and can therefore be separated. A detector measures what leaves the column; the chromatogram plots the measured signal against time.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><a href=\"https:\/\/www.waters.com\/nextgen\/us\/en\/education\/primers\/beginner-s-guide-to-liquid-chromatography\/how-does-high-performance-liquid-chromatography-work.html\" target=\"_blank\" rel=\"noopener\">Source: Waters \u2014 how HPLC and detectors work<\/a><\/p>\n\n\n\n<p class=\"wp-block-paragraph\">With UV detection, the signal depends on the wavelength selected. Also consider how well the peaks are separated. Compounds leaving the column at almost the same time can produce overlapping peaks. A single prominent peak is therefore not, by itself, a complete identification of a peptide.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Lees verder over <a href=\"\/hoe-leest-u-een-chromatogram\/\">hoe u een chromatogram beoordeelt<\/a>, met aandacht voor assen, pieken en de grenzen van de interpretatie.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"lc-ms\" style=\"margin-top:48px;margin-bottom:16px;font-size:clamp(18.959px, 1.185rem + ((1vw - 3.2px) * 0.863), 30px);line-height:1.25\">What does LC-MS add?<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">LC-MS couples the separation to mass spectrometry. The mass spectrometer measures the mass-to-charge ratio of ions, written as m\/z. A peptide can carry multiple charges, so different m\/z signals may belong to the same peptide. The laboratory takes those charges into account when comparing the results with the expected molecular mass.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><a href=\"https:\/\/www.waters.com\/nextgen\/lt\/en\/library\/application-notes\/2024\/mass-directed-isolation-of-synthetic-peptides-using-the-waters-sq-detector-2.html\" target=\"_blank\" rel=\"noopener\">Source: Waters \u2014 peptide analysis with UV and mass detection<\/a><\/p>\n\n\n\n<p class=\"wp-block-paragraph\">A matching mass supports identification, but it is not the same as establishing the complete amino acid sequence. Fragmentation analysis, such as MS\/MS, can provide additional structural information. Ask which identity checks were actually performed; the label LC-MS alone does not give the full answer.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><a href=\"https:\/\/www.waters.com\/nextgen\/ca\/en\/applications\/biopharma-and-pharma\/protein-therapies\/peptide-mapping.html\" target=\"_blank\" rel=\"noopener\">Source: Waters \u2014 peptide mapping and structural information<\/a><\/p>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"zuiverheid-inhoud\" style=\"margin-top:48px;margin-bottom:16px;font-size:clamp(18.959px, 1.185rem + ((1vw - 3.2px) * 0.863), 30px);line-height:1.25\">What does this tell you about purity and quantity?<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">An HPLC purity percentage may be based on the area of the assigned peptide peak relative to the peaks included in the calculation. Check which calculation and detection method were used. That percentage does not automatically describe the entire contents of the vial: water and salts, for example, are not accounted for in the same way in a UV peptide analysis.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">A report showing 99% chromatographic purity therefore does not, by itself, prove that a vial contains the number of milligrams stated on its label. Purity and measured quantity are separate measurements.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><a href=\"https:\/\/www.sigmaaldrich.com\/DE\/en\/technical-documents\/technical-article\/protein-biology\/protein-quantitation\/explanation-of-peptide\" target=\"_blank\" rel=\"noopener\">Source: Sigma-Aldrich \u2014 peptide purity and peptide content<\/a><\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Both HPLC and LC-MS can form part of a quantitative assay. This requires an appropriate measurement approach, such as calibration with standards containing known amounts. Look in the report for the quantitative result, its unit and the basis of the calculation. A graph or method name does not replace those details.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><a href=\"https:\/\/openlab.help.agilent.com\/2.8\/en\/mergedProjects\/Glossary\/Glossary.htm\" target=\"_blank\" rel=\"noopener\">Source: Agilent \u2014 quantitation and calibration<\/a><\/p>\n\n\n\n<p class=\"wp-block-paragraph\">In <a href=\"\/identiteit-zuiverheid-gemeten-inhoud\/\">identiteit, zuiverheid en gemeten inhoud<\/a> leest u waarom deze resultaten afzonderlijke vragen beantwoorden.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"rapport-controleren\" style=\"margin-top:48px;margin-bottom:16px;font-size:clamp(18.959px, 1.185rem + ((1vw - 3.2px) * 0.863), 30px);line-height:1.25\">Five checks for your report<\/h2>\n\n\n\n<ol class=\"wp-block-list\"><li>Check that the product, variant and batch number match your documentation.<\/li><li>Check which technique and detector were used; do not assume additional tests were performed.<\/li><li>Keep reported identity, purity and quantity separate.<\/li><li>Check the units, test date and any explanation of discrepancies.<\/li><li>Ask the laboratory or supplier for clarification if a result is unclear.<\/li><\/ol>\n\n\n\n<p class=\"wp-block-paragraph\">This guide helps you read a report; it is not an assessment of a specific Novogin batch. Do not infer a sterility result or suitability for human use from an HPLC or LC-MS label. Draw conclusions only about properties that were actually tested and reported.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"verder-lezen\" style=\"margin-top:48px;margin-bottom:16px;font-size:clamp(18.959px, 1.185rem + ((1vw - 3.2px) * 0.863), 30px);line-height:1.25\">Further reading and reports<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\"><a href=\"\/en\/hoe-leest-u-een-coa\/\">Read also: how do you read a COA?<\/a><\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Controleer ook <a href=\"\/analyserapport-actuele-batch-controleren\/\">of het analyserapport bij de actuele batch hoort<\/a>.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><a href=\"\/en\/coas\/\">Browse available reports in the batch library<\/a><\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Would you like to compare product specifications alongside this explanation? <a href=\"\/en\/product-category\/research-peptides\/\">Explore research peptides for professional research<\/a>.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">This knowledge base explains analytical documentation for laboratory research. It does not provide advice on dosing, administration or medical use.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"bronnen\" style=\"margin-top:48px;margin-bottom:16px;font-size:clamp(18.959px, 1.185rem + ((1vw - 3.2px) * 0.863), 30px);line-height:1.25\">Bronnen<\/h2>\n\n\n\n<ul class=\"wp-block-list\"><li><a href=\"https:\/\/www.waters.com\/nextgen\/us\/en\/education\/primers\/beginner-s-guide-to-liquid-chromatography\/how-does-high-performance-liquid-chromatography-work.html\" target=\"_blank\" rel=\"noopener\">Waters \u2014 How Does High Performance Liquid Chromatography Work?<\/a>: scheiding, detectoren en chromatogrammen.<\/li><li><a href=\"https:\/\/www.waters.com\/nextgen\/lt\/en\/library\/application-notes\/2024\/mass-directed-isolation-of-synthetic-peptides-using-the-waters-sq-detector-2.html\" target=\"_blank\" rel=\"noopener\">Waters \u2014 Mass-Directed Isolation of Synthetic Peptides Using the Waters SQ Detector 2 (2024)<\/a>: massa-ladingverhoudingen en meerdere ladingstoestanden.<\/li><li><a href=\"https:\/\/www.waters.com\/nextgen\/ca\/en\/applications\/biopharma-and-pharma\/protein-therapies\/peptide-mapping.html\" target=\"_blank\" rel=\"noopener\">Waters \u2014 Peptide Mapping and Analysis<\/a>: structuurinformatie in een peptide-mappingworkflow; dit is niet automatisch dezelfde test als ieder LC-MS-rapport.<\/li><li><a href=\"https:\/\/www.sigmaaldrich.com\/DE\/en\/technical-documents\/technical-article\/protein-biology\/protein-quantitation\/explanation-of-peptide\" target=\"_blank\" rel=\"noopener\">Sigma-Aldrich \u2014 Peptide Sample Amount Determination<\/a>: peptidezuiverheid en peptidegehalte.<\/li><li><a href=\"https:\/\/openlab.help.agilent.com\/2.8\/en\/mergedProjects\/Glossary\/Glossary.htm\" target=\"_blank\" rel=\"noopener\">Agilent OpenLab CDS 2.8 \u2014 Glossary<\/a>: raadpleeg <strong>Calibration<\/strong> en <strong>Calibration standard<\/strong> voor de relatie tussen instrumentrespons en een bekende hoeveelheid.<\/li><\/ul>","protected":false},"excerpt":{"rendered":"<p>What do HPLC and LC-MS tell you about a peptide? Understand separation, mass detection, purity and measured quantity.<\/p>","protected":false},"author":2,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[45],"tags":[],"class_list":["post-4979","post","type-post","status-publish","format-standard","hentry","category-kennisbank"],"blocksy_meta":{"styles_descriptor":{"styles":{"desktop":"","tablet":"","mobile":""},"google_fonts":[],"version":7}},"_links":{"self":[{"href":"https:\/\/novogin.com\/en\/wp-json\/wp\/v2\/posts\/4979","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/novogin.com\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/novogin.com\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/novogin.com\/en\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/novogin.com\/en\/wp-json\/wp\/v2\/comments?post=4979"}],"version-history":[{"count":5,"href":"https:\/\/novogin.com\/en\/wp-json\/wp\/v2\/posts\/4979\/revisions"}],"predecessor-version":[{"id":5606,"href":"https:\/\/novogin.com\/en\/wp-json\/wp\/v2\/posts\/4979\/revisions\/5606"}],"wp:attachment":[{"href":"https:\/\/novogin.com\/en\/wp-json\/wp\/v2\/media?parent=4979"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/novogin.com\/en\/wp-json\/wp\/v2\/categories?post=4979"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/novogin.com\/en\/wp-json\/wp\/v2\/tags?post=4979"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}