HPLC Method Development

1 Day Course

HPLC method development training course for the experienced chromatographer. 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. Chromatographic modelling software-based exercises are included to give further understanding.

We recommend that course numbers are limited to 20 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.

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

Objectives

Establishing method objectives
Literature searching: What is known? What needs to be known?

Sample Preparation

Sample clean up
Analyte extraction
SPE explained
Mobile phase selection
Optimising for sample type/application

System choices

How to choose the appropriate injector/detector
Typical operating conditions
Developing and optimising injection conditions
Mobile phase flow & band broadening
(Van Deemter)
Modes of chromatography

Choosing a Column and Mobile Phase

Choosing the correct phase
Computer-based tools for column choice
Effects of column geometry
Review of modern stationary phases
Isocratic vs. Gradient operation
Theory and development of eluent gradients

Optimisation Strategies

Capacity factor, Efficiency, Resolution, Selectivity
Resolution equation
Step-by-step guide for logical method development
Example method developments

Quantitation & System Characterisation

Single and multi-level calibration
Internal standards
System suitability testing
Introduction to validation