Determination of sugars in honey using HILIC separation and RI detection

Importance of the Topic

Honey quality and authenticity are critical for consumer safety and market regulation. Accurate quantification of its major sugars enables differentiation between natural honey and adulterated or substitute products.

Objectives and Overview of the Study

This study aimed to develop a robust hydrophilic interaction chromatography method with refractive index detection for simultaneous determination of fructose, glucose, sucrose and maltose in various honey and honey substitute samples using a dedicated AZURA Sugar Analytical System.

Methodology and Instrumentation

Samples and calibration standards were prepared by dilution in water and acetonitrile. Separation was performed on a NH2 HILIC column maintained at 35 C using an isocratic eluent of 80% acetonitrile at a flow rate of 1.2 mL/min. A 5 μL injection volume was analysed with a refractive index detector at 20 Hz data rate. Calibration covered concentrations from 0.39 to 12.5 mg/mL with linearity coefficients above 0.999.

Main Results and Discussion

The method achieved baseline separation of all four sugars within 12 minutes, with high retention time repeatability. Natural bee and blossom honey contained around 37 to 39 g of fructose and 33 to 35 g of glucose per 100 g, with negligible sucrose and low maltose. Agave nectar was rich in fructose (54 g per 100 g) with no maltose. A commercial honey substitute showed very low fructose (1.24 g per 100 g) but high maltose (35.15 g per 100 g), indicating non-natural origin. Total sugar content of true honey samples exceeded 74%, while substitute profiles varied significantly.

Benefits and Practical Applications

• Rapid and reliable quantification of key sugars in honey products
• Effective quality control and authenticity testing in the food industry
• Support for compliance with international honey labeling standards

Future Trends and Opportunities

Increasing flow rates to shorten run times while monitoring column lifetime Integration with mass spectrometric detection for greater sensitivity Automation and miniaturization of sample preparation Expansion of the method to diverse food matrices for comprehensive sugar profiling

Conclusion

The developed HILIC-RI method using the AZURA system provides efficient separation and quantification of major sugars in honey and substitutes. Characteristic sugar patterns enable clear distinction between natural and adulterated products, supporting industry-wide quality assurance.

References

1 AID Zucker Sirupe Honig Zuckeraustauschstoffe und Süßstoffe Nr 1157
2 Honigverordnung vom 16. Januar 2004 BGBI I S 92
3 Codex Alimentarius Commission GB18796-2005 2005