UPLC-MS Analysis of Mono-, Di- and Oligosaccharides Using ACQUITY UPLC BEH Amide Columns

Importance of the topic

Rapid and accurate analysis of mono-, di- and oligosaccharides is vital in food science, pharmaceuticals and biotechnology for quality control, process monitoring and metabolic studies.

Study objectives and overview

This application brief evaluates a UPLC-MS method using an ACQUITY UPLC BEH Amide column for simultaneous separation and detection of nine saccharides ranging from monosaccharides to maltoheptaose. The aim is to achieve high resolution and sensitivity within a short analysis time.

Applied methodology and instrumentation

A 2.1×50 mm, 1.7 µm BEH Amide column was operated at 35 °C with a 5-minute gradient from 80 % to 50 % acetonitrile (Mobile Phase A) and acetone/water (Mobile Phase B), both containing 0.1 % NH4OH. The flow rate was 0.17 mL/min and injection volume 0.7 µL. Electrospray ionization in selective ion recording (SIR) mode targeted m/z 179.2 for hexoses, 341.3 for disaccharides and 503.4–1151.8 for higher oligosaccharides.

Instrumentation used

• Waters ACQUITY UPLC system with ACQUITY TQD mass spectrometer
• ACQUITY UPLC BEH Amide column (2.1×50 mm, 1.7 µm)

Main results and discussion

The method achieved baseline separation of all nine saccharides within 5 minutes, with symmetric peak shapes and retention time RSDs below 1 %. SIR detection provided selective sensitivity, enabling clear distinction between isobaric compounds and quantitation at sub-microgram levels. The short cycle time combined with simple sample preparation supports high sample throughput.

Benefits and practical applications

• High-speed separation reduces analysis time and solvent consumption
• Robust ESI-MS detection ensures specificity for complex mixtures
• Minimal sample preparation simplifies workflow in QA/QC and research labs

Future trends and potential applications

Advancements may include extending this approach to larger and branched oligosaccharides, coupling with high-resolution mass spectrometry for structural elucidation, and integrating automated sample handling for glycomics studies and industrial monitoring of carbohydrate-based processes.

Conclusion

The combination of ACQUITY UPLC BEH Amide chromatography and TQD mass spectrometry offers a fast, sensitive and reproducible platform for comprehensive saccharide profiling, demonstrating strong potential for routine applications in multiple fields.

References

No external references were provided in the source document.