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Understanding the HILIC Separation Method in LC


Hydrophilic interaction liquid chromatography, or HILIC, is a powerful mode of liquid chromatography (LC)that uses polar stationary phases and high-organic initial mobile phase conditions for the separation of polar analytes. Join us as we explore HILIC separations.

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In this series of Restek Tips, we are going to discuss the method development for HILIC-based LC methods. The first thing we’ll look at is the separation mechanism of HILIC. This is a separation mode using polar stationary phases. Because the stationary phase is polar, HILIC is well suited for the analysis of polar molecules that have limited or no retention on typical reversed phases. The mobile phases are similar to those used in reversed-phase LC. However, in HILIC separation, the initial condition is under highly organic mobile phase to generate a water-rich layer on the surface of the polar stationary phase.

Because polar analytes are soluble in the water layer, they can interact with the polar stationary phase and be retained in multiple ways including hydrogen bonding, dipole-dipole interaction, and ion exchange. With differential distribution between stationary phases, water layer, and mobile phases, the mixture of polar compounds can be selectively eluted from the HILIC column. And as the mobile phase has high content of organic solvent, the detection sensitivity is significantly increased when a mass spectrometer is used as the detector. So, if you think HILIC might be something you want to try, in the next few videos, we will show you the tips that can greatly help you develop your own HILIC methods.

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