Abstract:

Based on hydrophilic interaction liquid chromatography (HILIC), a method for the separation and
determination of stevioside, rebaudiodside C and rebaudiodside A in stevia with strong anion-exchange column was
established. According to the structural features and physical properties of the adsorbent and adsorbate, the separation
mechanism of polar components in stevia was elucidated preliminarily. We speculated that the possible mechanism might
be hydrogen bond and dipolar interaction. Compared with the national standard, the number of chromatographic peaks
increased and the resolution of major substances revealed an obvious improvement. As a result, good linear correlations
for stevioside, rebaudiodside C and rebaudiodside A were achieved in the concentration ranges of 0.056–5.600,
0.032–3.200 mg/mL and 0.204–24.000 mg/mL, and the linear correlation coefficients (R2) were 0.9991, 0.9989 and 0.9992,
respectively. Recovery rates of three components at low, middle and high concentration levels were 95.8%–101.1%, with
relative standard deviations less than 1.00% (n = 6). The method is convenient, sensitive, accurate and reproducible. It can
provide a reliable way of determining polar components in stevia.

Key words: hydrophilic interaction liquid chromatography, strong base anion-exchange column, stevia, polar component