Food Analysis

Profiling galactosyl oligosaccharides using HPAE-PAD

This application note demonstrates the Thermo Scientific Dual Eluent Generation Cartridge (EGC) capability and performance for profiling galactosyl oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD).

Find out more about the HPAE-PAD analysis of  galactosyl-oligosaccharide-containing samples >>

ExampleCoverGoal
To demonstrate Dual Eluent Generation Cartridge (EGC) capability and
performance for profiling galactosyl oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD).

Introduction
Thermo Scientific™ Dionex™ “Just Add Water” electrolytic eluent generation technology is an essential part of Reagent-Free™ Ion Chromatography (RFIC™) systems. This technology ensures excellent reproducibility and accuracy in eluent preparation. Thermo Fisher Scientific offers a new operating mode for RFIC systems called Dual EGC mode. This option can replace the manual preparation of the sodium hydroxide/sodium acetate (NaOH/NaOAc) eluent gradients required for analyzing complex carbohydrates.

This study assesses the performance of the Dual EGC mode for separating complex carbohydrates using
the Thermo Scientific™ Dionex™ CarboPac™ PA200 1 mm column.

Experimental
Full details of the experimental conditions can be found by downloading the application note.

Results and Discussion
Gradient methods using an isocratic pump
Uncharged oligosaccharides are typically separated on a Dionex CarboPac PA200 3 mm column set using a sodium acetate gradient in an eluent containing either 100 mM or 150 mM sodium hydroxide. In Dual EGC mode, eluent generators methanesulfonic acid (MSA) and potassium hydroxide (KOH) cartridges, are installed in series to generate KOH/KMSA eluents required for analyzing complex carbohydrates by HPAE-PAD. This extension of RFIC technology now enables analysts to run gradient methods using an isocratic pump. The user now has the flexibility to modify or change the eluent concentrations anytime through the Chromeleon CDS software.

Conclusion
Our study demonstrated excellent performance of Dual EG cartridge technology when applied to the HPAE-PAD analysis of galactosyloligosaccharide-containing samples using a Dionex CarboPac PA200 1 mm column. The method achieved great separation using an eluent gradient generated using Dual EGC mode on a Thermo Scientific Dionex ICS-6000 system.

Dual EGC mode enables the analyst to run gradient methods using an isocratic pump, offering improved reproducibility, eliminating manual preparation of eluents, and minimizing pump maintenance because the pump only needs to pump water, thus maximizing instrument uptime. Dual EGC mode shows excellent performance for HPAE-PAD of complex carbohydrates.

Find out more about the HPAE-PAD analysis of  galactosyl-oligosaccharide-containing samples >>

 

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