Course Description

This course is an introduction to the principles of HPLC analytical technique. This course is delivered to master's students at University College London School of Pharmacy. The course explains the basic principles of HPLC separation; it demonstrates how to use the instrument and software, provide guidance for method development in reversed-phase HPLC and explains the various types of chromatography based on the retention mechanism. It is intended to beginners but practised HPLC users may find a benefit to go through the course as well to deepen their understanding of how the separation works.

Director, Honorary Professor at UCL School of Pharmacy

Klara Valko

Leaving GSK after 22 years of service, she is now founder director of Bio-Mimetic Chromatography Ltd providing data analysis, biomimetic HPLC method development and measurements, in vivo drug distribution modelling and various courses for pharmaceutical companies. She is an Honorary Professor to the UCL School of Pharmacy since 2004; leading the Physchem/ADME module for the Drug Discovery MSc course. She has developed biomimetic HPLC methods for high throughput measurements of serum albumin, glycoprotein and phospholipid binding. She invented the Chromatographic Hydrophobicity Index (CHI) for the fast determination of a compound’s lipophilicity using generic gradient reversed phase chromatography. She has built mathematical statistical models based on measured properties to predict human in vivo drug distribution, such as, volume of distribution, DRUGeff max and tissue/plasma partition. She has contributed to the Drug Efficiency Index (DEI) concept and dose estimation based on DEI. She has published over 100 papers that have achieved over 3000 citations. She has written a book on “Physicochemical and Biomimetic Properties in Drug Discovery; Chromatographic Techniques for lead optimization” published by Wiley in 2014. Currently, she is working on the second edition of the “Handbook of Analytical Separations” Volume 1 “Separation Methods in Drug Synthesis and Purification” published by Elsevier.  

Course curriculum

  • 1

    HPLC fundamentals

    • Introduction to the course

      FREE PREVIEW

    • HPLC course handout (download, print and make notes during the lecture videos)

    • Basic principles of HPLC, its importance in the pharmaceutical industry

    • The chromatographic separation process, why peaks are broadening, which parameters influence efficiency

    • Test your understanding (basic principles of the chromatographic separation)

    • Test your understanding (efficiency of the chromatographic system the causes of peak broadening)

    • The various types of chromatography based on the retention mechanism: Adsorption (normal phase) chromatography

    • Test your understanding (adsorption/normal phase chromatography)

    • Principles of reversed phase chromatography. Isocratic and gradient elution. How do the mobile phase pH and organic solvent concentration influence the separation?

    • Test your understanding (reversed phase chromatography)

    • Practical HPLC, start up Agilent 1100 and Chemstation software. Create method and sequence files.

    • Test your understanding (setting up an HPLC Analysis)

    • Principles of reversed phase HPLC method development DryLab demonstration of the effects of conditions on the separation.

    • Method development strategies

    • Test your understanding (method development principles in reversed phase chromatography)

    • Method validation and description of an HPLC protocol. Example for changing the selectivity of the chromatographic system.

    • Method development and trouble shooting

    • Test your understanding (method development & trouble shooting)

    • Principles of ion-exchange, size-exclusion, chiral chromatography and bio-chromatography

    • Test your understanding (method validation and types of chromatography)

    • Discussions

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