Validation of UHPLC-ESI-MS/MS Method for Determining Steviol Glycoside and Its Derivatives in Foods and Beverages

The aim of this study was to validate a method for determining nine types of steviol glycoside and its derivatives in food and beverage products, using ultrahigh-performance liquid chromatography tandem mass spectrometry with electrospray ionization (UHPLC ESI MS/MS). The performance characteristics of the analysis method were determined along with their suitability for the intended use. Coefficient of determination (R²) calibration curves from 0.2 to 1.0 mg L^-1 were in the ranges of 0.9911-0.9990, 0.9939-1.0000 and 0.9973-0.9999 for a beverage, yogurt and snack, respectively. Intra-day precisions in terms of percent relative standard deviation (% RSD) of concentration, at 0.2, 0.5 and 1.0 mg L^-1, for the beverage, yogurt and snack were lower than 15% (1.1-9.3%). At all concentrations, percentage recoveries were in the accepted range of 70-120%. For the matrix effect study, matrix-matched calibration was used for all compounds, obtaining a linear concentration range from 0.2 mg L^-1 to 1.0 mg L^-1. Almost all matrix-matched results presented as percentage recoveries were within the accepted range of 80-120%. The limit of detection (LOD) for steviol glycosides ranged from 0.003 to 0.078 μg g^-1, while the limit of quantitation (LOQ) ranged from 0.011 to 0.261 μg g^-1. These results indicate that the modified test method can be applied to determine the presence of steviol glycoside and its derivatives in a wide range of sample matrices.