Optimization and validation of HPLC–DAD method for simultaneous analysis of sweeteners, preservatives, and caffeine in sugar-free beverages

Abstract In this study, the optimization and performance evaluation of a high-performance liquid chromatography with diode array detection (HPLC–DAD) method for simultaneous determination four sweeteners (acesulfame-potassium, saccharin, aspartame, rebaudioside A), two preservatives (sodium benzoate potassium sorbate), caffeine in sugar-free drinks are presented. The separation was carried out using gradient elution acetonitrile phosphate buffer (12.5 mM, pH = 3.3) on reversed-phase column. Under optimized conditions, allowed specific selective all target analytes less than 9 min. developed HPLC–DAD validated demonstrated excellent linearity (all analytical curves showed R 2 ≥ 0.9995), satisfactory accuracy (recovery values ranging between 94.1 99.2% real samples), repeatability (intra- inter-day relative standard deviations were ≤ 2.49%). Overall, 69 products available Hungarian market successfully tested applied method.