An analytical method was established for the detection of stevia components in natural sweetner preparation by thin layer chromatography (TLC) on silica gel. Stevia components were developed with chloroform-methanol-water (30:23:4) and detected by spraying the chromatograms with anisaldehyde-sulfuric acid. Five stevia components yielded distinct green spots. The detection limits for these stevia compounds by this method were 0.1μg.High performance liquid chromatography (HPLC) was applied for the quantitative determination of stevioside and rebaudioside A among stevia components. One gram of natural sweetner preparation was dissolved in 20ml of 50% methanol solution and cleaned up by column chromatography on silica gel. Stevia components were eluted from the silica gel column with chloroform-methanol-water (30:40:15). Stevioside and rebaudioside A in the eluate were analyzed quantitatively by HPLC. The analytical conditions for HPLC were as follows: column, Unisil Q-NH2; mobile phase, acetonitrile-water (83:17); detector, 210nm. The limits of detection by HPLC were 0.1μg of stevioside and 0.2μg of rebaudioside A.Fourteen commercial products were analyzed. Stevioside, rebaudioside A, B and C and steviolbioside were detected, and quantitative measurements indicated the presence of 0.47 to 37.9% stevioside and 0.26 to 11.9% rebaudioside A.
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HIROKADO et al. (1980) studied this question.
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