One of the most widely used method for quantification of complex carbohydrates is high-performance anion-exchange chromatography coupled with pulsed amperometric detection (HPAE-PAD). HPAE-PAD allows complex carbohydrate analysis by direct detection, i.e. no sample derivatization is required. A new operating mode for HPAE-PAD systems, called Dual Eluent Generation Cartridge (Dual EGC) mode, is available to support the analysis of complex carbohydrates. This mode of operation replaces the manual preparation of the sodium hydroxide/sodium acetate (NaOH/NaOAc) eluent gradients required for analyzing complex carbohydrates by HPAE-PAD. This technology ensures excellent reproducibility and accuracy in eluent preparation. In Dual EGC mode, eluent generators in series produce methanesulfonic acid (MSA) and potassium hydroxide (KOH) to generate KOH/KMSA eluents used for complex carbohydrate separation. Our study demonstrated good performance of this technology when applied to the analysis of complex carbohydrates including inulin, galacto-oligosaccharides (GOS), bovine fetuin glycans, and N-linked oligosaccharides from human serum IgG and human α-1-acid glycoprotein using a Dionex™ CarboPac™ PA200 1 mm column. Dual EGC enables the users to run the gradient methods using an isocratic pump, offers improved reproducibility, eliminates manual preparation of eluents, and minimizes pump maintenance because the pump only needs to pump water, thus maximizing instrument uptime. Eluent generation of MSA and KOH eliminates the potential risk of system contamination due to the usage of lower quality sodium acetate.

HPAE-PAD analysis of complex carbohydrates using dual eluent generation cartridge

256th National Meeting (2018)

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