Development of HPLC-PDA method for steviol glycosides analysis in food matrices

A high-performance liquid chromatography (HPLC) method was developed and validated for the analysis of 13 steviol glycosides (SGs) in food matrices. HPLC conditions with a 40-min run and a C18 column (250 mm × 4.6 mm, 5 µm) successfully separated 13 SGs. The regression coefficients ranged from 0.9995 to 0.9999. The limit of detection and limit of quantitation were 0.09-0.64 and 0.27-1.93 mg/kg, respectively. Rebaudioside A, stevioside, rebaudioside F, and rebaudioside C were selected for method validation, and the food matrices were categorized as beverages, fermented milk, and snacks. The intra-and interday precisions of the selected food matrices were 1.93-3.91 and 2.13-5.92%, respectively. The matrix effects were 5.43-6.88% (beverages), 13.29-15.57% (fermented milk), and 9.63-14.32% (snacks). Interlaboratory validation was conducted to confirm the reproducibility of the method. The developed HPLC method was successfully applied to quantify SGs in 130 commercially available processed foods in Korea.

The online version contains supplementary material available at 10.1007/s10068-026-02242-5.