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9. Quantitative Determination of
Steviosides by High Performance Thin Layer Chromatographyϯ
Meruga Ramalýngeswara Rao, Tummala Ramakrýshna, Sukala Kannababu And
Gottumukkala V. Subbaraju*
Laila Research Centre, Unit I, Phase
III, Jawahar Autonagar, Vijayawada –520 007, India
E-mail:
subbarajugottumukkala@hotmail.com
Abstract:
A simple, precise and rapid high
performance thin layer chromatographic (HPTLC) method has been
developed for the quantitative determination of four steviosides,
namely rebaudioside A, rebaudioside C, stevioside and dulcoside A in
Stevia rebaudiana leaves and extracts. The separation was
carried out on a pre-coated with silica gel 60F–254 on aluminium
plates as the stationary phase, eluted with ethyl acetate-ethanol-water
(13:3:2) as mobile phase. A good linear relationship between peak
areas and concentrations was obtained over the range of 0.5 – 3.0 µg /
spot for the four steviosides. The method was validated for precision
and recovery. The spike recoveries were within 98.05 – 100.54%. The %
RSD values of the precision were in the range of 0.76 – 2.42%.
Keywords: HPTLC,
rebaudioside A, rebaudioside C, stevioside, dulcoside A, Stevia
rebaudiana.
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