Organizing co mmittee

DEFINITION OF STEVIOSIDE IN Stevia rebaudiana SAMPLES 
BY HIGH PERFORMANCE CHROMATOGRAPHY 
 
D.K. Mutalova, R.A. Botirov, G.E. Saidova, A.Z. Sadikov, Sh.Sh. Sagdullayev 
 
S.Yu. Yunusov Institute of the Chemistry of Plant Substances Academy of sciences of the 
Republic of Uzbekistan st. Mirzo-Ulugbek, 77, 100170 Tashkent,
e-mail: [e-mail] 
Diabetes mellitus is the most acute medical and social problem in almost all 
countries of the world. The search for evidence-based approaches to the prevention of 
diabetes continues. 
Stevia rebaudiana
leaves, leaf extracts are used both for the 
prevention of diabetes mellitus and in the complex therapy of the disease. The main 
group of biologically active compounds in stevia raw materials are diterpene glycosides. 
Sweet diterpene glycosides were found in all aerial parts of Stevia: stevioside, 
rebaudiosides A, B, C, D and E, which have the ability to enhance insulin secretion, 
enhance the effect of insulin on cell membranes, stabilize glucagon secretion and blood 
sugar levels.
The object of the study is the aerial part of 
Stevia rebaudiana
growing in Uzbekistan. 
Several samples of plant materials were studied by numerical indicators. 
Name of indicator 
Characteristic 
Appearance 
The finished raw material consists of yellow-green, 
green and dark green parts of the plant 
Moisture content,% 
6,11 
5,98 
6,25 
6,15 
5,95 
Total ash,% 
7,0 
6,5 
7,3 
6,8 
6,8 
Organic impurities, % 
1,88 
0,98 
1,5 
0,85 
1,2 
Mineral impurities,% 
0,76 
0.55 
0,67 
0,70 
0,70 
And also developed a method for identifying the complex of glycosides in stevia 
leaves by spectrophotometry and high-performance liquid chromatography. 
The developed method for the quantitative determination of stevioside using high 
performance liquid chromatography (HPLC) was validated. This technique is simple 
and well reproduced. The developed method was validated according to the following 
indicators: specificity, linearity, accuracy, limit of detection and limit of quantitation. 
The analysis was carried out on an Agilent 1200 3D LC System chromatograph with 
a diode array detector in isocratic mode, using a LUNA NH2 250×4.6 mm 5 µm 
column, at a wavelength of 210 nm. As a comparison, standard samples of Stevia and 
Rebudioside A were used.
Fig. 1. Chromatogram of 
Stevia rebaudiana
raw material extract 
The metrological characteristics of the method are calculated. The developed 
technique is successfully used to study the stage-by-stage control of stevioside 
production. 

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Poster presentation 
94