A Validated HPLC-RID Method for Quantification and Optimization of Total Sugars: Fructose, Glucose, Sucrose, and Lactose in Eggless Mayonnaise

Mayonnaise is an oil-in-water emulsion containing 70–80% finely dispersed droplets of oil in a continuous phase of water. Since mayonnaise has a sour and acidic taste, its sugar profile is barely noticed and thus often disregarded. However, today, there are various variants of mayonnaise available on the market; hence, it is crucial to understand their mono- and disaccharide profile, in order to determine the precise total sugar composition. The traditional methods of sugar analysis available, such as titration, can only quantify sucrose and are unable to differentiate between mono- and disaccharides. The aim of this study was to develop and validate a method for the quantification of total sugars, including fructose, glucose, sucrose, and lactose, in eggless mayonnaise, using a high performance liquid chromatography refractive index detector (HPLC-RID). Sugars were separated on an amino column with an oven temperature of 35 °C, using an isocratic solvent system consisting of a 75:25 v/v mixture of acetonitrile and HPLC water, at a 0.9 mL/min flow rate with RID. Method validation was performed for the linearity, specificity, precision, accuracy, LOD, LOQ, and robustness. A linearity for total sugars, with a regression coefficient of 0.9998, was obtained within the range of 0.05024 to 10.048 mg/mL. The relative standard deviation was less than 2.0% for the intra-day and inter-day precision. The accuracy was found to be 96.78–108.88% using a three-level recovery method. The LOD and LOQ were also found to be suitable. The samples used in this study contained 0.24–10.32% total sugars. The sucrose value obtained matched the label claim of the products and no significant differences were observed between results in a paired sample t-test. This showed the applicability of the proposed method for analyzing the sugar profile in a finished product. Routine analysis of total sugars in eggless mayonnaise and similar finished products can thus be performed using this technique, which was found to be simple, rapid, and reproducible.