HPLC
IndustriesFood & Agriculture
ManufacturerShimadzu
Importance of the Topic
Monosaccharides and disaccharides are ubiquitous carbohydrates that significantly influence the taste, nutritional value, and shelf stability of beverages. Rapid and reliable quantification of these sugars is critical for product development, quality assurance, regulatory compliance, and consumer safety. Traditional methods often suffer from extended run times and limited compatibility with gradient elution, prompting the need for faster, more versatile analytical approaches.
Objectives and Study Overview
This study aimed to develop and validate a high‐speed method combining hydrophilic interaction liquid chromatography (HILIC) with evaporative light scattering detection (ELSD) for simultaneous separation and quantification of five common saccharides—fructose, glucose, sucrose, maltose, and lactose—in soft drinks. The method was applied to standard mixtures and real beverage samples to assess performance metrics such as resolution, linearity, repeatability, and throughput.
Methodology and Instrumentation
Separation was achieved on a ULTRON AF-HILIC-CD column (100 mm × 3 mm, 2 µm) using a gradient of 10 mmol/L ammonium acetate (A) and acetonitrile (B), with B held at 85 % for 0–0.5 min, reduced to 82 % over 0.5–3 min, then returned to 85 % at 5.01 min. The flow rate was 0.8 mL/min at 45 °C. ELSD-LT III detection employed the Wide function for automatic sensitivity optimization, a 40 °C drift tube, 350 kPa nitrogen nebulizer gas, and a 4 s filter. Samples were filtered (0.2 µm) and diluted 20-fold with water/acetonitrile (50:50).
Main Results and Discussion
The five saccharides were baseline separated within five minutes with retention time RSDs below 0.05 % and peak area RSDs under 3.6 %. Calibration curves plotted on double-logarithmic scales exhibited excellent linearity (r2 > 0.999). Analysis of two commercial soft drinks yielded precise concentrations (e.g., fructose 2,920 mg/L and 3,448 mg/L) with reproducibility generally below 3 % RSD for major analytes. The Wide function obviated manual sensitivity adjustments across varying sample concentrations.
Benefits and Practical Applications
- High sample throughput: complete saccharide profile in under five minutes.
- Gradient compatibility: HILIC gradient elution enables resolution of analytes with diverse polarities.
- Robust detection: ELSD with automatic sensitivity tuning simplifies method transfer and routine use.
- Quality control: suitable for beverage QC laboratories requiring rapid sugar profiling.
Future Trends and Opportunities
Further development may include extension to oligosaccharides and sugar alcohols, coupling with mass spectrometry for structural elucidation, and miniaturized flow systems for reduced solvent consumption. Integration with automated sample processing and data analytics could enhance throughput in industrial and research settings.
Conclusion
The HILIC-ELSD method described provides a fast, reproducible, and gradient‐compatible approach for simultaneous determination of common mono‐ and disaccharides in soft drinks. Its high resolution, linearity, and minimal sample preparation make it a valuable tool for routine beverage analysis.
References
- K. Matsuoka and N. Iwata. High Speed Analysis of Monosaccharides and Disaccharides in Soft Drinks by ELSD-LT III. Shimadzu Application Note L572, First Edition, September 2020.
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