HPLC Method Development Incorporating Quality by Design

1 Day Course

Chromacademy eLearning

HPLC method development training course incorporating a Quality by Design (QBD) approach. The course provides a step-by-step approach to all the crucial aspects of method development, including information gathering, mode of chromatography, development of mobile phase systems, column choice, principles of ionisation/suppression, optimisation of important chromatographic parameters (R, α, k’ and N), isocratic and gradient operation and detector choice and optimisation. Each aspect is discussed in detail with relevant examples used to demonstrate theoretical principles.


  • We recommend that course numbers are limited to 10 so that each delegate gets the opportunity to ask questions and fully participate in tutorial exercises 
  • As the training is delivered on-site, we can design the course material to suit your specific needs
  • Customisable written assessments are available if required


Course Overview | Download course details »

This one day course provides a step-by step approach to method development for HPLC using Quality by Design (QBD) principles. 

All sections are accompanied by participative tutorial exercises using interactive chromatgraphic modelling software simulations for increased understanding of critical concepts.

Who is this course for

This course is designed for the more experienced chromatographer with a good working knowledge of HPLC separations. Attendance of the Fundamental HPLC and HPLC Troubleshooting and Maintenance training courses is not compulsory but is advantageous.

Previous knowledge

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.

What you will learn

  • Setting method development objectives
  • Sample preparation
  • System choices
  • Choosing a column and mobile phase
  • Optimisation strategies
  • Quantitation and system characterisation

Course Outline

Introduction to QBD approach to method development

Combined resolution equation (k', N and alpha) with reference to practical method development process

Critical method parameters

  • Selectivity
  • Method robustness

Troubleshooting common problems relating to the sample

 Mobile phase considerations

  • Ionisation, pH and pKa
  • Buffers - role, types and molarity
  • Flowrates
  • Organic modifier 

Choosing a Column

  • Physical dimensions (length, ID, pore size)
  • Particle size, particle type, UHPLC
  • Temperature
  • Endcapping
  • Bonded silica phases - pH and mobile phase restrictions
  • Reversed phase ligand chemistry
  • Column classification tests (Tanaka, PQRI, HSM etc.)


Specificity, peak purity and detection

Gradient and isocratic elution

Prelimanary retention time scouting

Troubleshooting retention time issues

Second stage optimisation - optimising selectivity choices

Linearity, range and sensitivity

Methods of quantitative calibration