analysis of charged N-glycansAn earlier blog post on this topic, HILIC HPLC Stationary Phase Speeds Separation of Glycans (1), (link to blog post) featured two application notes plus a scientific poster using one of our latest mixed mode HILIC HPLC glycan columns (Thermo Scientific GlycanPac AXH-1 column) which provides the simultaneous separation of glycans by charge, size, and polarity. Soon after the release of the post, I received an email that there was another application note featuring the very same column that I had missed! Hence, here I am pleased to feature the third application using this HILIC HPLC glycan column.

Application 20754, Separation of 2AB Labeled N-Glycans from Bovine Fetuin on a Novel Mixed-Mode Stationary Phase, (downloadable PDF), describes hydrophilic interaction liquid chromatography (HILIC) and weak anion exchange mixed-mode chromatography method for the analysis of fluorescently labeled N-glycans released from proteins (bovine fetuin).

The advantage of using this particular HILIC HPLC glycan column (Thermo Scientific GlycanPac AXH-1 column) is that it provides the accurate quantification of glycans based on charge, which cannot be achieved with any other HPLC or UHPLC columns on the market! The column has been designed and tested for use with LC-fluorescence detection and LC-MS applications using volatile aqueous buffers (e.g., ammonium acetate or ammonium formate) and acetonitrile.

Other key features of this particular column include the used of high-purity spherical silica as substrate, availablitiy in 1.9 μm and 3.0 μm sizes for UHPLC and standard HPLC applications. For more details on the technical specifications of the column, do check out GlycanPac AXH-1 Column Datasheet (downloadable PDF).

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The method was developed on our rapid-separation HPLC system (Thermo Scientific Dionex UltiMate 3000 RSLC Analytical LC system) equipped with one of our fluorescence detectors (Thermo Scientific Dionex FLD-3400RS Fluorescence Detector). The analysis time was shortened from 40 min to 20 min.

This method was developed by scientists in our Sunnyvale Applications Lab in California.

Do check out our growing library of chromatography biopharma applications on this blog.

If you have any questions on this column or type of analysis, do enter your questions in the Comments box below; our experts look forward to hearing from you.