Development and validation of an RP-HPLC method for the simultaneous determination of artificial sweeteners, preservatives and caffeine in non-alcoholic beverages

抄録

Abstract
Consumption of a wide variety of beverages has increased considerably in modern diets. These products frequently contain additives such as artificial sweeteners, preservatives, and caffeine to enhance flavor, stability, and functional properties. However, due to the potential adverse health effects associated with excessive intake of these compounds, the development of reliable analytical methods for their determination is of considerable importance. Therefore, the aim of this study was to develop and validate a rapid reversed-phase high performance liquid chromatography (RP-HPLC) method coupled with ultraviolet diode-array detection (UV-DAD) for the simultaneous determination of acesulfame K, sodium saccharin, caffeine, aspartame, sodium benzoate, and potassium sorbate in non-alcoholic beverages. Optimal chromatographic separation was achieved on a Poroshell 120 EC-C18 column using isocratic elution with a mobile phase consisting of methanol and phosphoric acid solution (pH = 3.8) in a volume ratio of 20:80, at a flow rate of 1.0 mL/min and a constant column temperature of 25 °C. Detection was performed at 195, 230, 260, and 275 nm. Under the optimized chromatographic conditions, the run time was less than 4.5 minutes. The validated method demonstrated excellent linearity ( R ² > 0.999), satisfactory intra-day and inter-day precision (RSD = 0.02 – 2.94 %), and good accuracy, with recoveries ranging from 91.11 to 108.99 %. The developed method was successfully applied for the simultaneous determination of the target analytes in commercially available non-alcoholic beverages. Compared with previously reported methods, the proposed method provides shorter analysis time and simple sample preparation, making it suitable for routine quality control analysis.