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HPLC Method Development

1 Day

For the experienced chromatographer, this course provides a comprehensive, process-driven approach to the development of an optimal and robust HPLC method.

The course includes a detailed exploration of each critical parameter within a typical method, then builds this into step-by-step process for method development.  We apply relevant theory to underpin a practical approach which maps out the optimal route to a commercial quality method. Importantly we look at building robustness into each stage of development using a Quality by Design approach, highlighting many common problems and how we develop these out.

Video Overview


Video Walkthrough


●      Setting method development objectives

●      Sample preparation

●      System choices

●      Choosing a column and mobile phase

●      Optimising strategies

●      Quantitation and system characterisation


  • Design specification
  • What is QBD?
  • Building in robustness

Influencing Resolution

  • Retention factor
  • Efficiency
  • Selectivity

Sample Preparation

  • Influencing sample solubility
  • Choosing a diluent

Mobile Phases

  • Sample polarity
  • Organic modifiers
  • pH control
  • Flowrates


  • Physical dimensions
  • Particle type, sizes and UHPLC
  • Temperature
  • Chemically modified silica
  • Reversed phase ligands
  • Hydrophobic subtraction model classification

Gradient Elution

  • Isocratic or gradient
  • Optimising k’
  • Effect of each gradient parameter

Building a Practical Process

  • Sample preparation
  • Retention
  • Screening for alpha
  • Chromatographic optimisation
  • Linearity, range and sensitivity
  • Pre-validation

This course is designed for the more experienced chromatographer with a good working knowledge of HPLC separations. Completion of Understanding HPLC and HPLC Troubleshooting training courses would be advantageous.

Delegates should have a good knowledge of chromatography and experience as HPLC users. Some experience in method development is recommended. A good grounding in chemistry is also beneficial.

Understanding HPLC

HPLC Troubleshooting

Analytical Method Validation

Forced Degradation

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