Determination of 5-hydroxymethylfurfural (5-HMF) in molasses sample by GC-MS: application of different derivatization strategies

Food processing contaminants, particularly compounds such as 5-hydroxymethylfurfural (5-HMF) formed during thermal treatment, pose a significant concern for food safety. Since 5-HMF has been reported to exhibit genotoxic, mutagenic, and potentially carcinogenic effects, its accurate and reliable determination in food and biological samples is of critical importance. In this study, the analysis of 5-HMF was performed using Gas Chromatography-Mass Spectrometry (GC-MS), and different derivatization strategies were evaluated. The primary objective was to overcome the limitations of conventional analytical techniques and to develop a more sensitive and selective approach. Among the derivatization reagents tested, MSTFA resulted in weak signals and was therefore unsuitable for quantitative analysis. In contrast, PFPA reagent provided short retention time, stable ion production, and strong signal intensity, making it the most successful option for method development. A calibration curve was established using PFPA-derivatized 5-HMF standards within the range of 6,25–125 ppm, yielding a high linear correlation (R² = 0,9975). In a real food matrix, grape molasses, the 5-HMF level was determined to be 7028,7 ppm. This concentration was considerably higher than those reported in the literature. The discrepancy is likely attributable to the high sensitivity of the PFPA derivatization method as well as the production and storage conditions of the sample.