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. 2025 May 7;9(1):66.
doi: 10.1038/s41538-025-00431-w.

Rapid and high-throughput analysis of 5-hydroxymethylfurfural in honey by MALDI-MS with in-situ tetra (4-aminophenyl)-porphyrin derivatization

Zhi Sun #  1 , Fangfang Wang #  1 , Peipei Zhou  1 , Di Chen  2   3 , Lihua Zuo  4
Affiliations

Rapid and high-throughput analysis of 5-hydroxymethylfurfural in honey by MALDI-MS with in-situ tetra (4-aminophenyl)-porphyrin derivatization

Zhi Sun et al. NPJ Sci Food. .

Abstract

5-Hydroxymethylfurfural (HMF), a product of the Maillard reaction and caramelization in honey, serves as a crucial marker of food quality and thermal processing, underscoring the importance of regulatory surveillance for consumer safety. This study introduces a method for rapid, high throughput and eco-friendly detection of HMF, using derivatization in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 5,10,15,20-Tetrakis-(4-aminophenyl)-porphyrin was employed for in-situ derivatization of aldehydes after extraction by ethyl acetate, enhancing detection sensitivity by shifting low-molecular-weight aldehydes to higher regions. The calibration curves for HMF determination exhibited outstanding linearity. The detection limit reached 0.347 mg/kg with good accuracy and precision. Variations in the content of HMF in honey during heating were also assessed, revealing the increased formation of HMF under high temperature and prolonged heating. The method's proficiency was evidenced through the detection of various aldehydes. This method represents an advance in rapid analysis for food safety, aligning with green chemistry principles.

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Conflict of interest statement

Competing interests: The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Schematic overview of the in-situ derivatization procedure using TAPP for the detection of aldehydes in MALDI-MS analysis.
Fig. 2
Fig. 2. Optimization of TAPP-based derivatization conditions performed by mixing derivatization solutions before spotting.
The effect of A different derivatization temperatures, B different derivatization times (0: only mixing directly). Optimization of TAPP-based in-situ derivatization conditions. The effect of C the concentration of TAPP and D the concentration of FA on the signal intensity.
Fig. 3
Fig. 3. Feasibility study of TAPP as a derivatization reagent.
MALDI-MS spectra of A the HMF standard solution (400.0 μg/mL), B the HMF-free solution following in-situ derivatization with TAPP, and C the HMF standard solution (400.0 μg/mL) following in-situ derivatization with TAPP. CHCA was utilized as the MALDI matrix.
Fig. 4
Fig. 4
Intra-day and inter-day precisions and accuracy of HMF detection in honey samples determined by the proposed TAPP-assisted MALDI MS method for samples spiked with low, medium, and high concentrations of HMF.
Fig. 5
Fig. 5
The changes of content of HMF in honeys over time and temperature.
Fig. 6
Fig. 6. The potential applicability of the TAPP-based in-situ derivatization strategy.
MALDI-MS spectra of A Perilla aldehyde standard solution (200.0 μg/mL), B Citral standard solution (200.0 μg/mL), C Citronellal standard solution (200.0 μg/mL), and D p-Coumaraldehyde standard solution (200.0 μg/mL), all following in-situ derivatization with TAPP. CHCA was utilized as the MALDI matrix.
Fig. 7
Fig. 7. The evaluation of method greenness.
A The Green Analytical Procedure Index (GAPI), B The Analytical GREEnness (AGREE), C Analytical Greenness Metric for Sample Preparation (AGREEprep), D Blue Applicability Grade Index (BAGI), and E Sample Preparation Metric of Sustainability (SPMS).
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