Reversed-Phase or HILIC? Optimizing Retention of Polar Analytes

Polar analytes vary in complexity and chemical properties, ranging from nucleotides to small polar acids, like citric acid. It can be difficult to know where to begin when developing methods. Many options exist to retain these compounds. The differences between the two major techniques, reversed-phase liquid chromatography (RPLC) and hydrophilic interaction liquid chromatography (HILIC), are considerable. Each has its own benefits and drawbacks. Additionally, within each separation technique, specialized stationary phases can provide unique selectivity or retentivity benefits, specifically for polar retention.

This webinar will highlight the two most common techniques employed to retain polar analytes. A discussion of reversed-phase polar retention columns will demonstrate how low ligand density phases and mixed mode columns can be used successfully followed by a look at HILIC and the effective stationary phases for successful separations. In addition, information about how MaxPeak Premier High-Performance Surfaces (HPS) technology can be used to improve polar analyte analysis will be provided. By understanding the different techniques and considering the different stationary phases and technologies available, method developers can find more suitable analysis conditions for their tricky polar analytes.

Presenter: Kenneth Berthelette (Senior Scientist, Waters Corporation Chemistry and Consumables Group)

Ken earned his Bachelor's of Science degree in Professional Chemistry from Bridgewater State University, Bridgewater Massachusetts, in 2010. He has been with Waters Corporation since March 2012 and has worked on several product launches including CORTECS columns, UPC2 Torus and Trefoil columns, and the Atlantis Premier BEH Z-HILIC column. He changed job functions in 2021 and now supports Chemistry Aftermarket initiatives. His focus on applications and customer education has led to several seminars and webinars on topics including method development, method modernization and transfer, HILIC separation techniques, and LC system troubleshooting.