Amide-Bonded BEH HILIC Columns for High Resolution, HPLC-Compatible Separations of N-Glycans

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Matthew A. Lauber, Stephan M. Koza, Jonathan E. Turner, Pamela C. Iraneta, and Kenneth J Fountain
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Application Notes
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Application Notes
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GST (Glycan Separation Technology)

In the following work, we demonstrate that BEH Glycan Columns packed with 1.7, 2.5, and 3.5 μm particle sizes afford scalability between glycan separations performed under UPLC and HPLC-compatible conditions. With method transfer accounting for differences in particle diameter (dp), we additionally show that noteworthy resolution can be achieved with columns packed with larger particles at HPLC-compatible pressures, albeit with an increase in analysis time compared to UPLC-based separations using BEH Glycan 1.7 μm columns. Lastly, this study demonstrates that highly similar glycan profiles and experimental glucose unit (GU) values are obtained, independent of the particle size or LC system (ACQUITY UPLC or Alliance HPLC) employed for a separation. The availability of these various BEH Glycan Columns should be of interest to laboratories looking for flexibility during method development or a facile means of transferring glycan separation methods between UPLC and HPLC instrumentation.

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