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The CHROMacademy Essential Guide Webcast:
Effective HPLC Troubleshooting

Thursday 19th April 2018,
8:00am PDT/ 11:00am EDT/ 4:00pm BST/ 5:00pm CEST


Sponsored by


Troubleshooting is an essential skill for all HPLC users.  By looking at the top 5 problems submitted to the CHROMacademy troubleshooter we will provide you with the fundamentals of troubleshooting which can be applied to any problem you encounter.  We will discuss troubleshooting problems with your separation and relate these to instrument, sample preparation and column issues enabling you to troubleshoot problems more effectively and avoid issues in the future.

Presented by

Dwight Stoll (Associate Professor of Chemistry, Gustavus Adolphus College).
Tony Taylor (Technical Director, Crawford Scientific).

Topics Covered

Essential HPLC troubleshooting strategies

  • Discuss common sources of the top 5 HPLC problems
    • Peak tailing
    • Peak splitting
    • Positive baseline drift
    • Peak fronting
    • Variable peak height/area
  • Outline strategies to confirm and isolate the problem
  • Explain how to solve the problem
  • Provide knowledge on how to avoid the problem in the future

Presenter Information »


Effective HPLC Troubleshooting.

This month’s webcast provided you with troubleshooting strategies which can be applied to any HPLC problem by looking at the five most common issues which are submitted to our interactive troubleshooter.  The CHROMacademy material provided in this month’s tutorial will give you further insight into how to effectively troubleshoot some of the most common problems encountered in HPLC.

Our Quick guides, CHROMacademy course content, and archived webcasts and tutorials will allow you to become a troubleshooting expert.

Watch the webcast

Quick Guides

HPLC Troubleshooting Guide – Peak Tailing

Tailing peaks create issues with resolution, quantitation (integration), and reproducibility.  Peak shape is often the controlling factor when optimizing complex separations, especially when components are present in very different concentrations.  Therefore, this Quick Guide will examine the top ways to mitigate peak tailing effects.

HPLC Troubleshooting Guide – Peak Tailing »


HPLC Troubleshooting Guide to Peak Splitting Problems

A step-by-step guide to using the CHROMacademy HPLC troubleshooter to solve the problem of peak splitting.

HPLC Troubleshooting Guide – Peak Splitting »


HPLC Troubleshooting Guide to Changes in Selectivity, Retention, and Resolution

Find out how our HPLC troubleshooter can be used to quickly solve the complex problem of changes in selectivity, retention, and resolution in a chromatogram.

HPLC Troubleshooting Guide to Changes in Selectivity, Retention, and Resolution »


Retention Time Variability in HPLC

This article discusses the common causes of retention time drift and how it can b avoided.

Retention Time Variability in HPLC »


Optimize and Troubleshoot Your HPLC System: Degassers, Pumps, and Autosamplers

Learn how HPLC instrument components function in order to correctly maintain and troubleshoot the system for maximum up-time and optimum chromatography.  This webcast will focus on the degasser, pumps, and autosampler. Poorly degassed solvents will cause noisy baselines and columns may be damaged; pumps which do not deliver the correct flow rate can result in drifting retention times; while an autosampler which does not deliver the correct sample volume will impact peak shape and quantitation. 

Optimize and Troubleshoot Your HPLC System: Degassers, Pumps, and Autosamplers »


UHPLC Troubleshooting – How U is Your UHPLC?

Since its inception UHPLC has gained popularity for enabling fast and efficient chromatography which allows the demands of modern day analyses to be met; namely increased sensitivity and higher throughput. However, moving from conventional HPLC systems to UHPLC is not without its problems and the requirement to adjust to a new way of working. Fundamental practices such as careful sample and solvent preparation and the use of appropriate fittings and tubing to reduce extra column volume and maximize peak efficiency will be discussed. We will also consider important principles such as pressure/selectivity effects, the practical implication of frictional heating, and instrument parameters such as system solvent wash out volume (as opposed to gradient dwell volume). Finally, initial conditions for method development in UHPLC and some simple, easy to use equations which can be utilized to facilitate method transfer from HPLC to UHPLC will be provided.

UHPLC Troubleshooting – How U is Your UHPLC? »

HPLC Troubleshooting Masterclass

Using real world chromatograms submitted by CHROMacademy members from many different application areas, this webcast will illustrate how to recognize problems and explain further tests and investigations necessary to properly qualify the issue.  We will offer advice on how the problem may be resolved to help you gain valuable insight into recognizing, characterizing and fixing problems in HPLC to improve your data quality and reduce instrument downtime.

HPLC Troubleshooting Masterclass »


Troubleshooting Your HPLC Chromatogram: Selectivity, Resolution, and Baseline Issues

This session examines the common causes of changes in selectivity, loss of chromatographic resolution and baseline issues – such as splitting, fronting, tailing and shouldering. Strategies for problem identification and calculation of critical chromatography performance indicators will also be discussed.

Selectivity, Resolution, and Baseline Issues »


HPLC Troubleshooting: Autosampler, Column, and Detector Issues

Practical troubleshooting and maintenance information associated with HPLC autosamplers, columns and detectors.  This session examines different autosampler designs, injection valve operations and common problems associated with modern HPLC autosamplers.  Sample solvent and injection volume effects on chromatography will also be discussed in detail.  We will consider column hardware design, materials of construction and associated issues – including minimizing dead volume and avoiding/troubleshooting column blockage issues, especially with the popular sub 2 µm column packing materials.  We will also investigate the importance of controlling the temperature of the column during analysis and the potential chromatographic effects of poor temperature control.  We will conclude by reviewing common HPLC detector hardware problems and the chromatographic issues associated with detector hardware and acquisition settings.   We will build a checklist of suggested maintenance operations as well as outlining common diagnostic tests.

HPLC Troubleshooting: Autosampler, Column, and Detector Issues »


HPLC Troubleshooting: Eluents and Solvent Delivery Systems

Practical troubleshooting and maintenance information around HPLC eluents and solvent delivery systems (pumps).  This session considers important practical concepts such as good practice for eluent preparation, filtration and degassing as well as common related issues such as retention time drift and selectivity changes.  We also explain the working principles of HPLC solvent delivery systems and highlight the components which are most susceptible to problems.  The webcast concludes by considering various troubleshooting tests and strategies as well as common maintenance operations for solvent delivery systems. A must see for anyone who uses HPLC equipment and who wants to improve their troubleshooting and instrument maintenance skills.

HPLC Troubleshooting: Eluents and Solvent Delivery Systems »

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Dwight Stoll
Associate Professor of Chemistry
Gustavus Adolphus College

Dwight Stoll received Bachelor of Science degrees in plant biology and biochemistry from Minnesota State University.  Following some time in industry working in separation science Dwight gained his PhD from the University of Minnesota working under the guidance of Prof. Peter Carr studying fast, comprehensive two-dimensional liquid chromatography. 

Dwight has been a faculty member at Gustavus Adolphus College since 2008, where he mainly teaches quantitative and instrumental analysis courses, in addition to directing a vibrant research program focusing on the development of rapid multidimensional liquid chromatography for both targeted and untargeted analysis in complex matrices, optimization of isocratic and gradient elution HPLC, and the characterization and role of selectivity in reversed-phase HPLC.

Dwight is the author or co-author of 35 peer-reviewed publications and two book chapters in the area of separation science. In 2009 Dwight was the winner of the John B. Phillips Award for contributions to multidimensional gas chromatography, and in 2011 he was the recipient of LCGC’s Emerging Leader in Chromatography Award. Most recently he was recently elected to the editorial advisory boards of LCGC Magazine and the separation science journal Chromatographia.

Tony Taylor
Technical Director, Crawford Scientific

Tony comes from a pharmaceutical background and has many years’ research and development experience in small molecule analysis and bio-analysis using LC, GC and hyphenated MS techniques.

He is actively involved in method development within the analytical services laboratory at Hall Analytical – which supplies contract research in Chromatography and Mass Spectrometry. He continues to research in novel separation technologies, chromatographic method development and structural characterisation, especially in the areas of Extractable and Leachable analysis and Bio-Chromatography.

Tony is the Technical Director of CHROMacademy and has spent the past 17 years as a trainer and developing on-line education materials in analytical chemistry techniques.