Chromatogram of monosaccharides and sugar alcohols - Effect of temperature

Importance of the Topic

Accurate separation and quantitative analysis of mono- and oligosaccharides is essential in quality control and research, given their roles in nutrition, pharmaceuticals and biotechnology. Analytical techniques that combine size exclusion and sodium-based ligand exchange offer improved resolution and retention, facilitating precise profiling of complex carbohydrate mixtures.

Objectives and Study Overview

This application note evaluates the performance of the Shim-pack SUR-Na column operated in size exclusion mode and sodium-based ligand exchange mode. The study assesses retention behavior and separation efficiency for a series of monosaccharides, disaccharides, trisaccharides and sugar alcohols across different column temperatures.

Methodology and Instrumentation

Water was used as the mobile phase at a constant flow rate of 0.6 mL/min, while column temperature was varied at 80 °C, 70 °C, 60 °C and 50 °C. Five mixtures (A–E) containing distinct sugar sets were injected at 20 μL. A refractive index detector recorded elution profiles, enabling comparison of retention times and peak resolution under each condition.

Instrumentation Used

• Liquid chromatograph: Nexera™ lite
• Analytical column: Shim-pack SUR-Na (250 mm × 7.8 mm I.D., 8 µm)
• Guard column: Shim-pack SUR-Na (G) (50 mm × 7.8 mm I.D., 8 µm)
• Mobile phase: Deionized water
• Flow rate: 0.6 mL/min
• Column temperature: 50–80 °C (10 °C increments)
• Injection volume: 20 µL
• Detector: Refractive index detector (RID)

Main Results and Discussion

Temperature strongly influenced chromatographic performance. At 80 °C, retention times decreased overall but critical pairs sometimes showed reduced resolution. Lower temperatures (50 °C) extended analysis time yet improved peak separation, particularly for isomeric monosaccharides and sugar alcohols. Size exclusion mode efficiently separated higher-molecular-weight oligosaccharides, while sodium-based ligand exchange enhanced selectivity among closely related monosaccharide isomers.

Benefits and Practical Applications

• Temperature flexibility allows optimization between analysis speed and resolution.
• Size exclusion mode provides rapid profiling of oligosaccharide fractions.
• Ligand exchange mode offers superior discrimination of structural isomers.
• Use of pure water as mobile phase simplifies operation and reduces solvent costs.

Future Trends and Opportunities

Growing demand for high-throughput carbohydrate analysis will drive innovations in column chemistries and detection methods. Coupling with mass spectrometry could enable structural elucidation, while miniaturized LC systems may facilitate real-time monitoring in bioprocessing and pharmaceutical production.

Conclusion

The Shim-pack SUR-Na column on a Nexera lite system delivers robust dual-mode separation of carbohydrates. Adjustable temperature control and complementary separation mechanisms support reliable, efficient analysis of diverse sugar mixtures in research and quality control settings.