Critical Evaluation of HPLC Methods

There are numerous factors that affect the quality of your HPLC separation; including mobile phase composition, temperature, ionic strength, pH, column dimensions, and stationary characteristics. However, what changes should we expect when these parameters are altered? Will retention improve if another solvent is used and would we expect selectivity to change? Will analytes elute in the same order if the mobile phase pH, ionic strength, additives or temperature are changed? In this Essential Guide we will evaluate the critical aspects of the HPLC process giving you a useful practical guide for optimizing HPLC variables which will result in improved and more robust chromatography.

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• Understand and evaluate each part of your HPLC method
• Understand how each of the factors that affect your HPLC separation can be used to alter analyte retention and selectivity
• Learn why and when you should use in-line filters or guard columns
• Understand how impurities, free silanol groups, or end-capping on the silica support material can impact on your HPLC separation – both positively and negatively
• Understand the common HPLC detectors, their advantages and disadvantages, and their application areas

CHROMacademy Content

• Reversed Phase (Partition) Chromatography
• Column Chemistry with Webcast!

Related Articles from LCGC Magazine

• How Reversed Phase LC Works
• A Three-Pronged Approach for Rapid HPLC Method Development
• Material Identification With Charged Aerosol Detection
• Elevated Temperatures in Liquid Chromatography, Part I: Benefits and Practical Considerations

CHROMacademy Webcasts / Essential Guides

• Developing Better Methods for Reversed Phase HPLC (Part 2)
• Mobile Phase Optimization Strategies for Reversed Phase HPLC
• Translating and Transferring HPLC Methods
• Understanding HPLC Column Characterization