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HPLC Troubleshooting and Maintenance class room course
1 Day Course | HPLC Level 2

A logical approach to HPLC Troubleshooting and Maintenance is fully explored within this one-day course. Commonly encountered problems and best practice are reviewed for all major system components including mobile phase, pump, injection system, column and detector. Examination of separation chemistry, interrogation of poor peak shape and retention problems are a fundamentally important part of the course, leading to increased awareness of troubleshooting principles.

  • We limit numbers to 20 per course so that each delegate gets the opportunity to ask questions and fully participate in tutorial exercises
  • When delivered on-site we can design the course material to suit your specific training needs
  • Customisable written assessments are available if required

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download course pdfThis one-day course introduces the fundamentally important concepts associated with troubleshooting HPLC analysis including hardware basics, separation issues, peaks shape and baseline issues and more.

All components of the HPLC system and associated problems are systematically studied. The use of chromatograms as diagnostic tools is explored in depth, including baselines, peak shape and retention time inconsistencies, spurious peaks, etc.

Suitable as a refresher for the more experienced analytical chemist or as an invaluable introduction to the technique for those with limited experience, this course provides an invaluable insight into HPLC principles and practice.

Who is this course for

This course is designed for those relatively new to HPLC, and ideally will follow-on from attendance on the fundamental HPLC training course.

Previous knowledge

No previous knowledge of chromatography is necessary, but basic experience of chemistry is beneficial.

What you will learn

  • How the chromatographic process takes place
  • The architecture of a modern HPLC system
  • Working principle and problems associated with each module of the HPLC system
  • How to use chromatograms to troubleshoot chromatographic problems
  • Best practices for column use and care
  • Best practice in system maintenance and performance evaluation

Approaches to Logical Troubleshooting

  • System overview
  • Component perspective
  • Symptomatic perspective
  • System maintenance records
  • Symptoms/Causes/Diagnosis and Solutions

Symptomatic Perspective: Baselines

  • Baseline spikes
  • Noisy baselines
  • Cycling baselines
  • Rising/falling baselines

Component Perspective

  • What to look for/what can go wrong with…
  • Autosamplers
  • Detectors: UV/RI/Fluorescence
  • Solvent delivery systems & mobile phase

Symptomatic Perspective: Peaks

  • No peaks
  • Fronting/tailing peaks
  • Split peaks/shoulders
  • Broad peaks
  • Ghost peaks
  • Retention stability
  • Loss of sensitivity
  • Correct integration methods


  • Installation and conditioning
  • Column chemistry
  • Efficiency loss
  • pH operating range/bleed
  • Proper column management
  • Loss of sensitivity


  • Maintenance schedules
  • Correct maintenance procedures for all system components
  • Column maintenance
  • Performance evaluation

Training Calendar

Click on a title below to download a detailed course description or click a date and book your course.

Can't find a suitable training course? Call 01357 522 961 or email us.

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Manual Injection Complete and Partial Loop Filling

Various options exist for the injection of a given volume of sample solution into the flow of mobile phase – most methods use a loop injector and a pre-determined filling regime. Traditional loop injectors can be filled in one of the two ways outlined below:

Complete Loop Filling - As the loop contains mobile phase, the sample solution introduced will necessarily mix and become diluted with the resident mobile phase whilst it is in the process of displacing it. Therefore, in order for the loop to be filled with homogeneous sample it should be overfilled between 2 and 5 times with the sample solution thereby eliminating any possible dilution effects. A 20 μl injection loop should be filled with between 40 and 100 μl of sample for example.

Partial Loop Filling - The partial loop filling technique is typically utilised when it is important to preserve valuable or limited sample, or if the correct sample loop is not available for the injection volume required. To obtain the best possible precision the volume of sample solution injected should be no greater than 50% of the injector loop capacity. A 100 μl loop should not be used for injections of over 50 μl for example.

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