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Describes systematic troubleshooting workflow, emphasizing gathering facts, checking simpler causes first, comparing expected vs. observed performance, documenting actions in instrument logs, and observing safe laboratory and electrical safety practices.",{"@graph":14,"@context":71},[15,34,54],{"@type":16,"itemListElement":17},"BreadcrumbList",[18,23,27,31],{"item":19,"name":20,"@type":21,"position":22},"https://docshare.wps.com","Home","ListItem",1,{"item":24,"name":25,"@type":21,"position":26},"https://docshare.wps.com/document/","Document",2,{"item":28,"name":29,"@type":21,"position":30},"https://docshare.wps.com/document/technology/","Technology",3,{"item":32,"name":10,"@type":21,"position":33},"https://docshare.wps.com/document/chromatography-successful-hplc-operation-a-troubleshooting-guide/672891/",4,{"url":32,"name":10,"@type":35,"image":36,"author":41,"headline":10,"publisher":44,"fileFormat":47,"inLanguage":8,"description":12,"dateModified":48,"datePublished":48,"encodingFormat":47,"isAccessibleForFree":49,"interactionStatistic":50},"DigitalDocument",{"url":37,"@type":38,"width":39,"height":40},"https://docshare.wps.com/thumbnails/chromatography-successful-hplc-operation-a-troubleshooting-guide/672891.png","ImageObject",300,407,{"name":42,"@type":43},"RuangKosong","Person",{"url":19,"name":45,"@type":46},"DocShare","Organization","application/pdf","2026-11-02",true,{"@type":51,"interactionType":52,"userInteractionCount":22},"InteractionCounter",{"@type":53},"ViewAction",{"@type":55,"mainEntity":56},"FAQPage",[57,63,67],{"name":58,"@type":59,"acceptedAnswer":60},"What is the recommended troubleshooting strategy for HPLC issues?","Question",{"text":61,"@type":62},"Use a five-step process: identify the problem, understand the possible cause(s), isolate the exact cause, rectify if able, then return to routine use or refer to maintenance management.","Answer",{"name":64,"@type":59,"acceptedAnswer":65},"How should troubleshooting be carried out to find the exact cause efficiently?",{"text":66,"@type":62},"Follow a systematic sequence: gather facts rather than theories, check simpler items first, compare obtained vs. expected performance, list possible causes, and work through them step-by-step, checking outcomes after each change.",{"name":68,"@type":59,"acceptedAnswer":69},"What safety practices must be observed before troubleshooting HPLC instruments or columns?",{"text":70,"@type":62},"Follow safe laboratory practices, know solvent chemical/physical properties and have MSDS available, and power down/unplug electrically powered instruments before removing covers. Wear appropriate eye protection when troubleshooting the detector with its cover removed.","https://schema.org",{"og:url":32,"og:type":73,"og:title":10,"og:site_name":45,"og:description":12},"article",{"robots":75,"canonical":32},"index,follow",{"doc_id":77,"site_id":7},672891,1793612261,{"code":4,"msg":80,"data":81},"success",[82,86,90,94,99,103,108,113,118,121,125],{"id":22,"doc_module":4,"doc_module_name":25,"category_name":83,"show_sort_weight":84,"slug":85},"Story & Novel",90,"story-novel",{"id":26,"doc_module":4,"doc_module_name":25,"category_name":87,"show_sort_weight":88,"slug":89},"Literature",80,"literature",{"id":33,"doc_module":4,"doc_module_name":25,"category_name":91,"show_sort_weight":92,"slug":93},"Exam",70,"exam",{"id":95,"doc_module":4,"doc_module_name":25,"category_name":96,"show_sort_weight":97,"slug":98},5,"Comic",60,"comic",{"id":100,"doc_module":4,"doc_module_name":25,"category_name":29,"show_sort_weight":101,"slug":102},6,50,"technology",{"id":104,"doc_module":4,"doc_module_name":25,"category_name":105,"show_sort_weight":106,"slug":107},7,"Healthcare",40,"healthcare",{"id":109,"doc_module":4,"doc_module_name":25,"category_name":110,"show_sort_weight":111,"slug":112},8,"Research & Report",30,"research-report",{"id":114,"doc_module":4,"doc_module_name":25,"category_name":115,"show_sort_weight":116,"slug":117},9,"Religion & Spirituality",20,"religion-spirituality",{"id":116,"doc_module":4,"doc_module_name":25,"category_name":119,"show_sort_weight":116,"slug":120},"World Cup","world-cup",{"id":122,"doc_module":4,"doc_module_name":25,"category_name":123,"show_sort_weight":122,"slug":124},10,"Lifestyle","lifestyle",{"id":126,"doc_module":4,"doc_module_name":25,"category_name":127,"show_sort_weight":95,"slug":128},19,"General","general",{"code":4,"msg":80,"data":130},{"doc_id":77,"user_id":131,"nickname":42,"user_avatar":132,"doc_module":4,"category_id":100,"category_name":29,"doc_title":10,"doc_description":12,"doc_content":133,"file_id":134,"file_url":135,"file_type":136,"file_size":137,"view_count":22,"is_deleted":4,"is_public":22,"is_downloadable":22,"audit_status":22,"page_count":138,"language":139,"language_code":8,"site_id":7,"html_lang":8,"table_of_contents":140,"faqs":141,"seo_title":142,"seo_description":12,"update_tm":78,"read_time":143},962090883219,"https://ap-avatar.wpscdn.com/davatar_276721f389ce27ea32af1340a28f341c","c h r o m a t o g r a p h y  \nSuccessful HPLC Operation  \nA Troubleshooting  \nGuide  \nPart of Thermo Fisher Scientific  \n2  \nTable of Contents  \n1. Introduction 4  \n 1.1 Purpose 4  \n 1.2 Content 4  \n2. Troubleshooting Strategy and Processes 5  \n 2.1 Strategy 5  \n 2.2 Troubleshooting Process 5  \n3. Troubleshooting Isolation and Corrective Action 6  \n3.1 Visual Inspection 6  \n3.2 Pressure 6  \n 3.2.1 High Pressure 7  \n 3.2.2 Low Pressure 8  \n 3.2.3 Fluctuating Pressure 9  \n3.3 Baseline Irregularities 10  \n 3.3.1 Non-Cyclic Noise – Fluid Path Problems 10  \n 3.3.2 Non-Cyclic Noise – Detector Electronics Problems 12  \n 3.3.3 Cyclic Noise – Detector Related Problems and Others 13  \n3.4 Changes In Chromatography 14  \n 3.4.1 Retention Time Changes from Injection to Injection 14  \n 3.4.2 Continually Increasing or Decreasing Retention Times 16  \n 3.4.3 Increasing or Decreasing to a New Constant Retention Time 17  \n 3.4.4 Abnormal Peak Shape 17  \n3.5 Qualitative Results 26  \n3.6 Quantitative Results 28  \n4. Good Laboratory Practice for HPLC 30  \n4.1 Preparation of Solvents 30  \n 4.1.1 Quality 30  \n 4.1.2 Buffers 30  \n 4.1.3 Filtration 30  \n 4.1.4 Degassing 31  \n4.2 Solvent Use 31  \n 4.2.1 Instrument 31  \n 4.2.2 Mobile Phase Properties 32  \n4.3 Changing Solvents 32  \n 4.3.1 Buffered Phase to Wash or Storage Phase 32  \n 4.3.2 Normal to Reversed Phase and Vice Versa 32  \n 4.3.3 General 33  \n4.4 System Plumbing and Fittings 33  \n 4.4.1 Cutting Tubing 34  \n 4.4.2 Fittings 35  \n 4.4.3 SLIPFREE™ Fittings 36  \n 4.4.4 Blocked Tubing 36  \n 4.4.5 Old and Leaking Fittings 37  \n4.5 Column Selection 37  \n4.6. Column Protection 38  \n 4.6.1 Guard Columns and Cartridges 38  \n 4.6.2 Pre-Column Filters 39  \n4.7 Column Operating Temperature 39  \n4.8 Sample Preparation 40  \n5. Reference Data 41  \n5.1 – 5.1.5 Chromatographic Performance Tests 41  \n 5.1.6 Band Spreading or Band Broadening 44  \n5.2 System Calculations 45  \n 5.2.1 Column Backpressure and Typical Operating Flow Rates 45  \n 5.2.2 Scaling Up or Scaling Down 46  \n5.3 Solvent Properties 47  \n5.4 Buffer Properties 48  \n5.5 Chromophore Detection Wavelengths 50  \n5.6 Column Cleaning and Regeneration 51  \n 5.6.1 Normal Phase Media 51  \n 5.6.2 Reversed Phase Media 51  \n 5.6.3 Anion Exchange Media 51  \n 5.6.4 Cation Exchange Media 51  \n 5.6.5 Protein Size Exclusion Media 51  \n 5.6.7 Porous Graphitic Carbon 52  \n 5.6.8 Polymeric Media with Metallic Counter Ions 53  \n5.7 Chemical Compatibility of Polymeric Tubing With Solvents 54  \nAppendices  \nAppendix A: Column Selection Guide 56 Appendix B: Solvent Miscibility Chart 59 Appendix C: Example Routine Maintenance Log 60 Appendix D: Example Problem Log 61 Appendix E: Example Column History Log 62 Appendix F: Example Instrument Use Log 63  \nReferences 68  \n3  \n4  \n1. Introduction  \n1.1 Purpose  \n1.2 Content  \nThis guide is intended as an aid in troubleshooting problems that occur in the day to day running of your HPLC system. It is also intended as a guide to best practice in systems that are free from problems.  \nIts primary aims are to provide a logical process when troubleshooting, to maximize system operation time and to promote good chromatographic practices.  \nWherever possible, diagrams or schematics are used for clarity. This enables the guide tobe used quickly and efficiently by operators with varying experience.“Quick Tips” at the end of each section are included for rapid troubleshooting and less experienced HPLC users.  \nEach troubleshooting chapter contains information on the most likely cause(s) of the observed problem and ways in which they can be rectified. Each table has the most probable cause of the observed fault as it’s first entry.  \nThe scope of this guide has been extended where necessary to include LC/MS column and instrument troubleshooting. The focus of the bulk of the causes of problems and their corrective action is the UV and other flow-through detectors.  \n2. Troubleshooting Strategy and Processes  \n2.1 Strategy  \nAny troubleshooting str","cbCaitflBhNqI7Yj","https://ap.wps.com/l/cbCaitflBhNqI7Yj","pdf",1268948,64,"English","# 1. Introduction\n## 1.1 Purpose\n## 1.2 Content\n# 2. Troubleshooting Strategy and Processes\n## 2.1 Strategy\n## 2.2 Troubleshooting Process\n# 3. Troubleshooting Isolation and Corrective Action\n## 3.1 Visual Inspection\n## 3.2 Pressure\n## 3.2.1 High Pressure\n## 3.2.2 Low Pressure\n## 3.2.3 Fluctuating Pressure\n## 3.3 Baseline Irregularities\n## 3.4 Changes In Chromatography\n## 3.5 Qualitative Results\n## 3.6 Quantitative Results\n# 4. Good Laboratory Practice for HPLC\n## 4.1 Preparation of Solvents\n## 4.2 Solvent Use\n## 4.3 Changing Solvents\n## 4.4 System Plumbing and Fittings\n## 4.5 Column Selection\n## 4.6 Column Protection\n## 4.7 Column Operating Temperature\n## 4.8 Sample Preparation\n# 5. Reference Data\n## 5.1 – 5.1.5 Chromatographic Performance Tests\n## 5.1.6 Band Spreading or Band Broadening\n## 5.2 System Calculations\n## 5.3 Solvent Properties\n## 5.4 Buffer Properties\n## 5.5 Chromophore Detection Wavelengths\n## 5.6 Column Cleaning and Regeneration\n## 5.7 Chemical Compatibility of Polymeric Tubing With Solvents","[{\"question\":\"What is the recommended troubleshooting strategy for HPLC issues?\",\"answer\":\"Use a five-step process: identify the problem, understand the possible cause(s), isolate the exact cause, rectify if able, then return to routine use or refer to maintenance management.\"},{\"question\":\"How should troubleshooting be carried out to find the exact cause efficiently?\",\"answer\":\"Follow a systematic sequence: gather facts rather than theories, check simpler items first, compare obtained vs. expected performance, list possible causes, and work through them step-by-step, checking outcomes after each change.\"},{\"question\":\"What safety practices must be observed before troubleshooting HPLC instruments or columns?\",\"answer\":\"Follow safe laboratory practices, know solvent chemical/physical properties and have MSDS available, and power down/unplug electrically powered instruments before removing covers. Wear appropriate eye protection when troubleshooting the detector with its cover removed.\"}]","Chromatography - Successful HPLC Operation - A Troubleshooting Guide | PDF",161]