Development and Validation of an Enzymatic Method To Determine Stevioside Content from Stevia rebaudiana

作者：Udompaisarn Somsiri; Arthan Dumrongkiet; Somana Jamorn^*

来源：Journal of Agricultural and Food Chemistry, 2017, 65(15): 3223-3229.

DOI：10.1021/acs.jafc.6b05793

• stevioside determination
• enzymatic method
• beta-glucosidase
• Stevia rebaudiana
• method validation
• stevioside

An enzymatic method for specific determination of stevioside content was established. Recombinant beta-glucosidase BT_3567 (rBT_3567) from Bacteroides thetaiotaomicron HB-13 exhibited selective hydrolysis of stevioside at beta-1,2-glycosidic bond to yield rubusoside and glucose. Coupling of this enzyme with glucose oxidase and peroxidase allowed for quantitation of stevioside content in Stevia samples by using a colorimetric-based approach. The series of reactions for stevioside determination can be completed within 1 h at 37 degrees C. Stevioside determination using the enzymatic assay strongly correlated with results obtained from HPLC quantitation (r(2) = 0.9629, n = 16). The percentages of coefficient variation (CV) of within day (n = 12) and between days (n = 12) assays were lower than 5%, and accuracy ranges were 95-105%. This analysis demonstrates that the enzymatic method developed in this study is specific, easy to perform, accurate, and yields reproducible results.

• 出版日期2017-4-19