Analysis of Predominant Steviosides in Stevia rebaudiana Bertoni by Liquid Chromatography/Electrospray Ionization-Mass Spectrometry

Stevia reabudiana is gaining popularity especially in making diabetic food supplements
in view of its non-nutritive glycosides. Steviosides profile in ex vitro and in vitro leaves
and callus cultures of S. rebaudiana were investigated by HPLC analysis and further
confirmed by MS using mass spectral fragmentation data obtained from LC-MS-ESI
studies. The leaf samples (4 weeks old) and callus cultures (2 weeks old) were used for
extraction and extracts were used for HPLC analysis. Predominantly, eight sweet diterpene
glycosides were detected. The highest stevioside content of 6.48% Steviolbioside and
0.099% Rebaudioside-A on a dry weight basis were found in ex vitro and in vitro leaves,
respectively. In callus and suspension cultures, the biosynthesis of steviosides was
observed except for steviolbioside. The LC-MS-ESI based detection of different steviosides
in Stevia cultures could be used for comprehensive qualitative evaluation of the samples.