Full optimization and validation of an hplc method for the quantitative analysis of total sugars in a soft drink

Abstract

Five HPLC methods were employed for the quantitative analysis of three natural sugars namely fructose, glucose and sucrose in soft drinks. HPLC-refractive index detector (RID)-AMINO proved to be the most suitable HPLC method to carry out the latter task. For the optimum separation and response of the natural sugars the best conditions employed were column oven temperature 30 oC, flow rate 0.1 mL/min, mobile phase ratio acetonitrile:water 75:25 and they were determined by studying all possible interactions among these three parameters. Full validation of HPLC-RID-AMINO was performed in terms of system suitability test, precision check, accuracy check and robustness.