5-Hydroxymethyl Furfural (HMF) Evaluation in Molasses Produced through Different Traditional Methods Using HPLC

Molasses is a widely consumed traditional food product obtained from various fruits and valued for its nutritional and sensory properties. Thermal processing during molasses production promotes Maillard reactions, which contribute to desirable color and flavor development but may also result in the formation of undesirable compounds such as 5-hydroxymethylfurfural (HMF). Elevated HMF levels are associated with high processing temperatures and prolonged storage periods and are considered indicators of food quality and safety. In this study, a validated high-performance liquid chromatography–photodiode array detection (HPLC–PDA) method was employed to determine HMF levels in grape, mulberry, and carob molasses produced by traditional methods. The method demonstrated excellent linearity (R² = 0.9993) within the calibration range of 6.25–50 µg/mL. The limits of detection (LOD) and quantification (LOQ) were determined as 5.93 µg/mL and 17.97 µg/mL, respectively. HMF was quantified at 26.24 µg/mL in carob molasses, while grape and mulberry molasses contained HMF levels below the LOQ. In addition, total phenolic content (TPC) and total flavonoid content (TFC) were evaluated. Carob molasses exhibited the highest TPC (748.59 mg/L) and TFC (270.64 mg/L), followed by mulberry and grape molasses. Statistical analysis revealed significant differences among molasses types (p<0.05). The findings indicate that traditional production practices may affect HMF formation. Regular monitoring of HMF and optimization of production conditions are important to ensure product quality and consumer safety.