TITLE:
Determination of Rebaudioside A and Stevioside in Leaves of S. rebaudiana Bertoni Grown in México by a Validated HPLC Method
AUTHORS:
Irma Aranda-González, Yolanda Moguel-Ordoñez, David Betancur-Ancona
KEYWORDS:
Stevia rebaudiana, Rebaudioside A, Stevioside, HPLC, Validation
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.6 No.11,
October
21,
2015
ABSTRACT: Stevia rebaudiana is a plant with high
sweetening capacity due to its content of glycosides, mainly stevioside and
rebaudioside A. Several techniques have been used to determine the concentrations
of glycosides in Stevia, although an
HPLC method is recommended by the FAO/WHO-JECFA. Varieties of Stevia have been recently grown in
Mexico, with no previous report of glycosides by a validated method. The aim of
this study was to validate an isocratic HPLC method for content determination
of main glycosides in the leaves of Stevia cultivated in Mexico. HPLC method was performed using a C18 column (250 mm ×
4.6 mm, 5 μm) and UV detector set at 210 nm. The mobile phase consisted of
32:68 (v/v) mixture of acetonitrile and sodium-phosphate buffer (10 mmol/L, pH
2.6), set to a flow rate of 1.0 mL/min. Rebaudioside A and stevioside were
determined in two Stevia varieties:
Morita II and Criolla, and also validation parameters were calculated. Rebaudioside
A content (g/100g) in Morita II was 15.15 ± 0.02 while stevioside was 3.97 ± 0.003;
in the case of Criolla they were 4.03 ± 0.01 and 8.80 ± 0.14, respectively (p