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Development and Validation of a Reversed-Phase High-Performance Liquid Chromatography Method for Routine Identification and Purity Assessment of High-Purity Steviol Glycoside Sweeteners

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journal contribution
posted on 12.02.2014, 00:00 authored by Tsion Bililign, Jeffrey C. Moore, Shane Tan, Allan T. Leeks
The widespread application of stevia-based sweeteners in food products has resulted in the need for reliable analytical methods for measuring the purity and identity of high-purity steviol glycoside ingredients. The objective of this research was to develop and validate a new reversed-phase separation method capable of separating and quantifying nine steviol glycosides present in typical high-purity stevia extract ingredients. Results of the study established the linearity of the method at a correlation factor of 1.000 for the two major components and other minor components of this food ingredient. Method accuracy values were in the range of 99.1–100.9%. The percent relative standard deviation for six independent assay determinations was 1.0%. The method was determined to be robust for minor changes in column temperature, initial acetonitrile content, flow rate, and wavelength. The validated high-performance liquid chromatography method was found to be suitable to be included by USP as a Food Chemicals Codex compendial standard for steviol glycosides.