A Novel HPLC Method for Rapid Determination of HMF in Dulce de Leche: A Potential Tool for Quality Control

A validated isocratic method utilizing reverse-phase high-performance liquid chromatography (RP-HPLC) was developed for the quantification of hydroxymethylfurfural (HMF) in five distinct commercially available Dulce de Leche samples that are sold in Turkish markets. The X-Terra RP-18 column, which has dimensions of 150 × 4.60 mm i.d. and a particle size of 5 μm, was selected for the stationary phase. The separation process was conducted isocratically by utilizing a methanol-water mixture (10:90, v/v) containing 1% TCA as the mobile phase. The separation was carried out at a temperature of 25 oC and a flow rate of 0.5 mL/min. The detection was performed at a wavelength of 284 nm. The methodology exhibited favorable selectivity and linearity, as evidenced by a correlation coefficient (r) exceeding 0.999. The obtained values for the limit of detection (LOD) and limit of quantification (LOQ) were 0.0019 μg/mL and 0.0059 μg/mL, respectively. Additionally, the recovery rate was determined to be 95.45%. Furthermore, it has been reported that the levels of HMF in commercially available Dulce de Leche originating from Turkey vary between 50.55 and 90.18 mg/kg for the total HMF content.