Some of the Learning Outcomes

This two-day course covers HPLC theory, troubleshooting and method development. Content can be tailored to the client’s requirements and a hands-on, practical element may be included if desired.

Topics include:

HPLC Theory

Important chemical concepts

  • Bond polarity
  • Hydrogen bonding
  • Acid-base theory


  • Definitions of basic terms
  • Plate number, separation factor, retention factor and resolution
  • The Van Deemter Equation

Mobile and stationary phases

  • Retention mechanisms
  • Normal phase, HILIC, aqueous normal phase, reversed phase, chiral separations, ion
  • chromatography and size exclusion chromatography
  • Effect of particle size on efficiency/UHPLC
  • Core-shell packings
  • Reversed-phase retention mechanisms
  • Strategies for the retention of ionized compounds


  • Isocratic, low-pressure and high-pressure mixing
  • Gradient delay volume

Sample introduction

  • Types of autosampler
  • Sample filtration

Column oven and column temperature effects


  • Detector properties
  • UV/visible (including diode array), fluorescence, refractive index, conductivity, electrochemical, light scattering, charged aerosol and mass spectrometric detectors


  • System suitability as a troubleshooting tool
  • History of method and previous problems
  • Connecting tubing
  • Pump priming and leak testing
  • Causes of leaks
  • Diagnosing pump, autosampler, column and detector faults
  • Column care

Method development

  • Analyte properties (acid/base properties, solubility and detectability)
  • Matrix properties
  • Sample matrix effects
  • Achieving adequate specificity
  • Injection solvent effects
  • Method performance requirements
  • Method performance and drug substance/drug product specifications
  • Sources of analytical variability
  • Typical specification limits
  • Sampling error
  • Sample preparation
  • Analyte extraction
  • Sample stability
  • Developing the separation
  • Regular separations (neutral and ionisable organic molecules)
  • Special cases
  • Choice of initial conditions
  • Stationary phase chemistries and retention mechanisms
  • Column temperature effects

Benefits of In-house Trainings

Get to know the Trainer

Mark Powell
Mark PowellDirector
Mark Powell Scientific Limited

Mark Powell is a Fellow of the Royal Society of Chemistry (RSC) with over twenty-five years’ experience as an analytical chemist. He is an expert in sampling and sample preparation, chromatography (liquid and gas), spectroscopy (atomic and molecular) and pharmaceutical dissolution testing.  Mark was Honorary Treasurer of the RSC’s Analytical Division and led a working group on continuing professional development until July 2016.

After completing a PhD in analytical chemistry, he worked at a senior level in a number of industrial laboratories. Mark joined Liverpool John Moores University’s School of Pharmacy and Chemistry in 1997 as a Senior Lecturer, teaching chromatography, mass spectrometry and sample preparation.  He was Programme Leader for the University’s MSc in Instrumental Chemical Analysis and was also active in research and consultancy.

In 2003, he joined the newly-formed Quay Pharmaceuticals, a contract research and manufacturing organisation specialising in early-stage drug development, where he set up the analytical facility and managed the analytical development programme. In 2010 Mark was appointed Quay’s Scientific Manager, becoming involved more generally with drug development programmes and establishing collaborations with a number of UK universities and instrument manufacturers. Mark left Quay Pharmaceuticals in September 2013 to set up Mark Powell Scientific and has enjoyed working with companies of all sizes around the world on a variety of training, consultancy and expert witness assignments.

If you wish to know more about his experience, do not hesitate to contact us.

Feel free to suggest the topics that you would like to learn about at our HPLC Training Course.

Request an Offer

Request detailed Agenda - HPLC Theory In-house Training

Who should attend?

This course is intended for analytical scientists working in pharmaceutical QC and development roles:

  • Analytical Scientist
  • QC Chemist/Senior QC Chemist
  • Development Chemist/Senior Development Chemist
  • Product Development Scientist
  • QC Manager
  • Analytical Development Manager

Our Pharmaceutical events were attended by these companies