Honey Analysis Made Easy

Importance of the Topic

Honey analysis is critical for ensuring food safety, protecting public health and preserving bee populations. Contaminants such as antibiotic residues, pesticide traces and plant‐derived toxins can pose significant risks to consumers and pollinators. Comprehensive screening for carbohydrates, vitamins and potential adulterants (e.g., corn syrup) also underpins quality control and authenticity verification.

Objectives and Study Overview

This compilation of application notes presents six targeted workflows for honey analysis:

• Quantification of sulfonamide antibiotics and trimethoprim by QuEChERS extraction and UHPLC‐MS/MS.
• Detection of neonicotinoid and phenylpyrazole pesticide residues using QuEChERS and UHPLC‐MS/MS.
• Assessment of Tripterygium wilfordii toxins (wilfordine, wilforine) by liquid–liquid extraction and LC‐MS/MS.
• Optimization of carbohydrate profiling via HPLC method scouting coupled to single‐quadrupole MS.
• Ultra‐high‐sensitivity analysis of water‐soluble vitamins using UHPLC with a high‐sensitivity PDA cell.
• Quantitative FTIR‐ATR screening of fructose, glucose and sucrose for honey authenticity.

Methodology and Instrumentation

Key methodologies and instruments include:

• Sample preparation: QuEChERS and dispersive SPE for antibiotics and pesticides; liquid–liquid extraction for nectar toxins; simple dilution for vitamin and FTIR analyses.
• Chromatographic systems: Shimadzu Nexera UHPLC (LCMS‐8050 and LCMS‐8060) for MS/MS workflows; Nexera SR with SPD‐M30A PDA for vitamins; HPLC method scouting system with single‐quadrupole MS for carbohydrate profiling; IRTracer‐100 FTIR with ATR accessory for sugar quantitation.
• Detection modes: Multiple Reaction Monitoring (MRM) in positive/negative electrospray ionization; full‐scan MS for scouting; photodiode array detection with extended optical path length; FTIR‐ATR spectra processed by PLS chemometrics.

Main Results and Discussion

Antibiotics:

• 15 sulfonamides and trimethoprim showed linear calibration (r² ≥ 0.990) over 0.5–500 pg on‐column.
• LOQs of 0.5 pg on‐column, recoveries 54–91% (two compounds < 20%).

Pesticides:

• LOQs down to 1 ng/kg for fipronil, 5–20 ng/kg for neonicotinoids.
• Honey samples contained low ppb levels, well below regulatory limits; method stable over 150 injections.

Nectar toxins:

• Wilfordine and wilforine LODs of 1.3 and 4.3 ng/L, LOQs of 2.7 and 9.0 ng/L.
• Recoveries 99–116%, r² > 0.999.

Carbohydrates (HPLC‐MS scouting):

• Automated evaluation of 6 columns and 16 mobile phases pinpointed an optimized gradient for 17 sugars.
• Seamless software control reduced method development time.

Vitamins:

• Using Nexera SR high‐sensitivity PDA, six water‐soluble vitamins were separated in < 3 minutes with enhanced S/N (7× improvement).

FTIR sugar assay:

• PLS calibration (r² ≥ 0.9986) for fructose, glucose, sucrose in 34 standards.
• Commercial honeys labeled “100% pure” showed elevated glucose/fructose ratios, indicating possible corn syrup adulteration.

Contributions and Practical Applications

These validated workflows enable laboratories to:

• Rapidly screen honey for antibiotics, pesticides and toxic plant metabolites at ultra‐trace levels.
• Optimize LC methods efficiently for complex carbohydrate mixtures.
• Perform high‐throughput quantitation of vitamins and sugars to assess nutritional quality and detect adulteration.
• Maintain robust, reproducible performance across large sample batches.

Future Trends and Potential Applications

Emerging directions include:

• Integration of high‐resolution MS and ion mobility for deeper compositional profiling.
• Miniaturized sample prep (µQuEChERS) and on‐site FTIR/near‐infrared handheld devices for rapid field screening.
• Machine‐learning algorithms for spectral deconvolution and authenticity prediction.
• Extension to pollen, propolis and other bee‐derived matrices for comprehensive environmental monitoring.

Conclusion

Combining advanced extraction techniques, automated chromatographic scouting, high‐sensitivity detection and chemometric analysis provides a powerful analytical platform for safeguarding honey quality and authenticity. These methods deliver the speed, sensitivity and reproducibility needed by research, QA/QC and regulatory laboratories.

References

• Clifford R. I. et al. Quantification of Natural Sugars in Baby Food Products by MID FTIR Spectroscopy. AN FTIR-1401, 2014.
• Jagdish T., Irudayaraj J. Quantification of Saccharides in Multiple Floral Honeys using FTIR-ATR. J Agric Food Chem, 2004;52(11):3237–3243.
• Tucker M. et al. FTIR Quantitative Analysis of Sugars and Lignin in Pretreated Softwood Residues. Appl Biochem Biotech, 2001;91–93:51–61.
• Cadet F., Offmann B. Direct Spectroscopic Sucrose Determination of Raw Sugar Cane Juices. J Agric Food Chem, 1997;45:166–171.
• Kramer R. Chemometric Techniques for Quantitative Analysis. Marcel Dekker, 1998.