Analysis of Saccharides Using Integrated HPLC and Size Exclusion-Ligand Exchange (Ca Type) Column

Importance of the Topic

Monitoring saccharide levels in food and beverages is critical for quality control, nutritional labeling, and managing health risks such as obesity and dental caries. Reliable, high-throughput methods for separating and quantifying both monosaccharides and sugar alcohols enhance analytical capabilities in food science and industrial settings.

Objectives and Study Overview

This study demonstrates the use of an integrated high performance liquid chromatography system coupled with a calcium-type size exclusion-ligand exchange column to achieve selective separation and quantification of sucrose, glucose, fructose, and sorbitol. The method aims to simplify mobile phase preparation by using water only while maintaining high selectivity and reproducibility.

Methodology and Instrumentation

• Column setup: Shim-pack SCR-101C analytical column (300 mm×7.9 mm I.D., 10 µm) with Shim-pack SCR-101C guard column (50 mm×4 mm I.D., 10 µm)
• Stationary phase: Calcium counterion for enhanced retention via metal-ligand exchange combined with size exclusion effects
• Mobile phase: Water at a flow rate of 1.0 mL/min
• Column temperature: Maintained at 80 °C to prevent anomer peak splitting
• Injection volume: 10 µL
• Detection: Differential refractive index detector (RID-20A) with positive polarity, cell temperature at 40 °C, response time 1.5 s

Key Results and Discussion

• Separation performance: Baseline resolution of sucrose, glucose, fructose, and sorbitol achieved within a 15-minute run
• Calibration linearity: Correlation coefficients of r2 ≥ 0.9999 for all four saccharides over 20 to 2000 mg/L
• Repeatability: Retention time relative standard deviation below 0.2% and peak area RSD below 1.2% for six consecutive injections at 50 mg/L
• Real sample analysis: Soft drink and energy drink samples diluted and filtered prior to analysis; quantified saccharide concentrations ranged from 1.8×104 to 9.2×104 mg/L
• Spike and recovery: Fortified samples at 500 mg/L yielded recovery rates between 86% and 99%

Advantages and Practical Applications of the Method

• Water-only mobile phase eliminates buffer preparation and reduces waste
• Integrated HPLC platform maintains compact footprint while supporting refractive index detection
• High selectivity for both monosaccharides and sugar alcohols supports food and beverage quality control
• Robust performance with excellent linearity, sensitivity, and reproducibility

Future Trends and Applications

Emerging trends include coupling water-based ligand exchange columns with mass spectrometry for enhanced compound identification, miniaturization for high-throughput screening, and expansion to complex matrices such as dairy and pharmaceutical formulations. Advances in green chromatography will further drive adoption of water-only mobile phases in routine analysis.

Conclusion

The integrated HPLC method using a Ca-type size exclusion-ligand exchange column provides a streamlined, reliable approach for simultaneous analysis of sucrose, glucose, fructose, and sorbitol. The water-only mobile phase, combined with refractive index detection, offers a practical solution for rapid and reproducible saccharide quantification in real samples.

References

1. World Health Organization. Guideline Sugars intake for adults and children. 2015.