Beaming steviol glycoside analysis into the next dimension

- First ultra-high performance liquid chromatography application separating nine steviol glycosides.
- Nine UHPLC columns tested.
- Three columns yield resolution of 2 or greater for stevioside and rebaudioside A.
- Sufficient retention of rebaudioside D on two columns.
- Total run time 16 min.

Nine state-of-the-art reversed phase (RP) columns for ultra-high performance liquid chromatography were tested for the separation of steviol glycosides. The main criteria were resolution of the critical peak pair rebaudioside A and stevioside and the retention time of rebaudioside D. Three columns yielded a resolution of 2 or more of the critical peak pair and two of them showed sufficient retention of rebaudioside D, namely 1.62 and 1.84 min corresponding to retention factors of 0.98 and 1.24. The separation of nine steviol glycosides was possible in 11 min with UV and MS compatible, buffer-free eluents at moderate temperature. The presented method is proposed to be adopted as a new official method.