Determination of Rebaudioside A and Stevioside in Leaves of S.rebaudiana Bertoni Grown in Mexico by a Validated HPLC Method  

作　　者：Irma Aranda-Gonzalez Yolanda Moguel-Ordonez David Betancur-Ancona 

机构地区：[1]Facultad de Medicina,Universidad Autonoma de Yucatan,Merida,Mexico [2]Instituto Nacional de Investigaciones Forestales,Agricolas y Pecuarias(INIFAP),Campo Experimental Mococha,Carretera Merida-Motul,Mococha,Mexico [3]Facultad de Ingenieria Quimica,Universidad Autonoma de Yucatan,Merida,Mexico

出　　处：《American Journal of Analytical Chemistry》2015年第11期878-885,共8页美国分析化学（英文）

基　　金：supported by Programa de Mejoramiento al Profesorado-PROMEP-SEP;Fondos Fiscales-INIFAP

摘　　要：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 < 0.001). The recoveries of fortified samples were 100% ± 10% and precision RSD was ≤6.27%. The criteria of validation showed accuracy, linearity (≥0.99), and precision;therefore, the determination of glycosides was performed with reliability.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 < 0.001). The recoveries of fortified samples were 100% ± 10% and precision RSD was ≤6.27%. The criteria of validation showed accuracy, linearity (≥0.99), and precision;therefore, the determination of glycosides was performed with reliability.

关 键 词：Stevia rebaudiana Rebaudioside A STEVIOSIDE HPLC VALIDATION 

分 类 号：R73[医药卫生—肿瘤]