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Evaluating the Adsorbed Water Layer on Polar Stationary Phases for Hydrophilic Interaction Chromatography (HILIC)

School of Pharmacy and Health Sciences, Fairleigh Dickinson University, Florham Park, NJ 07932, USA
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Separations 2019, 6(2), 19; https://doi.org/10.3390/separations6020019
Received: 26 January 2019 / Revised: 25 February 2019 / Accepted: 8 March 2019 / Published: 1 April 2019
The water-rich liquid layer immobilized on the surface of the polar stationary phases is critical to the retention of polar compounds in hydrophilic interaction chromatography (HILIC). Although the presence of the adsorbed water layer has been investigated and confirmed by multiple techniques, there is a lack of quantitative measures that can be easily determined and linked to chromatographic parameters. This study proposes a simple measure termed volume ratio (the ratio of the adsorbed water layer volume and the mobile phase volume) that can be easily determined using toluene elution volume. The volume ratio values measured using the proposed method indicate that the volume of the adsorbed water layer varies in a wide range in the stationary phases commonly used in HILIC separation. It was observed that the volume ratio increases with the acetonitrile content and ammonium acetate concentration in the mobile phase. In addition, increasing the column temperature had the effect of reducing the volume ratio and diminishing the adsorbed water layer. View Full-Text
Keywords: volume ratio; adsorbed water layer; stationary phase; salt concentration; column temperature volume ratio; adsorbed water layer; stationary phase; salt concentration; column temperature
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Guo, Y.; Bhalodia, N.; Fattal, B.; Serris, I. Evaluating the Adsorbed Water Layer on Polar Stationary Phases for Hydrophilic Interaction Chromatography (HILIC). Separations 2019, 6, 19.

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