PROFILING OF SACCHARIDES IN HONEY BY HILIC-MS

Significance of the Topic

Honey authenticity is a critical quality issue and saccharide profiling supports detection of adulteration and geographic origin claims through detailed carbohydrate composition analysis.

Objectives and Study Overview

This study aimed to develop and optimize a hydrophilic interaction liquid chromatography mass spectrometry (HILIC-MS) method for profiling and quantifying mono, di and trisaccharides in honey samples, enabling sensitive detection and differentiation from adulterants such as syrups.

Methodology and Instrumentation

Sample preparation involved dissolving honey and syrup samples in acetonitrile-water (1:1 v/v), filtration and dilution with internal standards of isotopically labeled fructose, glucose and sucrose. Chromatographic separation was performed on an XBridge BEH Amide XP column under a gradient of acetonitrile-water-methanol with diethylamine and guanidine hydrochloride additives. Detection used selected ion recording of chloride adducts under optimized cone voltage.

Instrumentation Used

• ACQUITY Arc chromatography system with XBridge BEH Amide XP column, 3.0 × 10 mm, 2.5 µm
• ACQUITY QDa single quadrupole mass detector in negative ESI mode at 600°C
• Mobile phases combining acetonitrile, water, methanol with 0.05% diethylamine and 500 ppb guanidine hydrochloride

Results and Discussion

Fifteen saccharide peaks including two monosaccharides, six disaccharides and seven trisaccharides were resolved with excellent separation efficiency and reproducibility. Retention times were matched to 25 screened standards for tentative identification, while additional unknown peaks indicated minor oligosaccharides previously unreported in honey. Profiles of honey differed significantly from syrup samples, demonstrating clear markers of authenticity. Quantitative calibration curves for fructose, glucose and sucrose showed linear response with high correlation coefficients.

Benefits and Practical Applications

• Enables robust authentication of honey by profiling minor saccharides and detecting adulteration
• Supports differentiation of botanical and geographical origin through unique carbohydrate fingerprints
• Provides quantitative assessment of major sugars aligned with Codex Alimentarius quality guidelines

Future Trends and Potential Applications

Future developments may include integration of high resolution MS for structural confirmation, expansion of oligosaccharide libraries for comprehensive profiling and implementation of automated workflows for high throughput quality control in the honey industry.

Conclusion

The optimized HILIC-MS approach using a BEH Amide XP column and QDa detector provides a powerful tool for detailed saccharide profiling in honey, enabling sensitive detection of adulteration and reliable assessment of origin.

References

1. Doner LW. The sugars of honey - a review. J Sci Food Agric 28:443-456, 1977
2. Codex Alimentarius Commission. Revised Codex standard for honey; Codex STAN 12-1981, Rev. 1 (1987), Rev. 2 (2001)
3. Da Costa Leite JM et al. Determination of oligosaccharides in Brazilian honeys of different botanical origin. Food Chemistry 70:93-98, 2000
4. Benvenuti M et al. Profiling mono and disaccharides in milk and infant formula using the ACQUITY Arc system and QDa detector. Waters Application Note 720005767en, 2016