Simultaneous Analysis of Monosaccharides and Oligosaccharides in Beer Using ELSD-LT III

Significance of the Topic

Beer flavor is shaped by the profile of mono- and oligosaccharides produced during starch saccharification. Precise quantification of these non-UV absorbing saccharides is crucial for brewer’s quality control, product development, and flavor consistency. Evaporative light scattering detection (ELSD) coupled with hydrophilic interaction liquid chromatography (HILIC) offers a versatile solution for simultaneous analysis of a broad saccharide range.

Objectives and Study Overview

• Develop a gradient HILIC method for simultaneous quantification of key beer saccharides.
• Assess separation of fructose, sucrose, malto-oligosaccharides (DP1–DP10) and iso-maltooligosaccharides (DP2–DP5).
• Demonstrate dynamic range extension to handle wide concentration variations without manual sensitivity adjustment.
• Apply the method to diverse commercial beer styles for compositional profiling.

Methodology and Instrumentation

The study used gradient elution on a HILIC column, transitioning from high acetonitrile to increased aqueous content to resolve saccharides by polarity and size. ELSD-LT III detection enabled universal response for non-chromophoric analytes and supported gradient operation.

Used Instrumentation

• Shimadzu i-Series LC-2050C HPLC system
• Shodex HILICpak VG-50 4E column (250 mm×4.6 mm, 5 µm)
• ELSD-LT III evaporative light scattering detector
• Mobile phases: Water (A) and Acetonitrile (B), flow rate 1.0 mL/min
• Column temperature 45 °C; ELSD drift tube 40 °C; N₂ nebulizer gas at 350 kPa; 4-second filter; wide gain setting

Results and Discussion

• Complete separation of ten malto-oligosaccharides and five iso-maltooligosaccharides within 50 minutes.
• Calibration curves plotted on logarithmic axes showed excellent linearity (R² >0.997 for all analytes).
• Repeatability: <0.2% RSD for retention times and <4% RSD for peak areas across five injections.
• Dynamic range extension function delivered reliable quantification across analyte concentration differences without manual sensitivity changes.
• Analysis of ten commercial beers revealed characteristic saccharide fingerprints: darker and craft beers exhibited elevated oligosaccharide levels; sugar-free beers showed trace amounts; alcohol-free beers contained higher monosaccharides.
• Spike-and-recovery in a lager matrix returned 90–110% recoveries, confirming accuracy in complex samples.

Practical Benefits and Applications

• Comprehensive saccharide profiling enhances flavor optimization and quality assurance in brewing.
• Gradient ELSD method streamlines analysis by eliminating multiple isocratic runs.
• Dynamic range extension simplifies routine quantification across broad concentration ranges.
• Suitable for R&D, QA/QC, and industrial monitoring of carbohydrate content.

Future Trends and Opportunities

• Coupling HILIC-ELSD with mass spectrometry for structural characterization of novel saccharide isomers.
• Automation and high-throughput workflows for large-scale screening in brewing innovation.
• Extension to other food and beverage matrices requiring non-chromophoric compound detection.
• Development of portable HILIC-ELSD systems for on-site process monitoring.

Conclusion

The gradient HILIC-ELSD method on the i-Series LC-2050C with ELSD-LT III achieves robust, precise, and accurate simultaneous quantification of beer mono- and oligosaccharides. Its excellent separation performance, dynamic range management, and applicability across diverse beer styles make it a valuable tool for both research and routine quality control in the brewing industry.

References

• Shimadzu Application News: Simultaneous Analysis of Monosaccharides and Oligosaccharides in Beer Using ELSD-LT III, First Edition: March 2024.