Somsiri UdompaisarnDumrongkiet ArthanJamorn SomanaMahidol University2018-12-212019-03-142018-12-212019-03-142017-04-19Journal of Agricultural and Food Chemistry. Vol.65, No.15 (2017), 3223-322915205118002185612-s2.0-85017654501https://repository.li.mahidol.ac.th/handle/20.500.14594/41571© 2017 American Chemical Society. An enzymatic method for specific determination of stevioside content was established. Recombinant β-glucosidase BT-3567 (rBT-3567) from Bacteroides thetaiotaomicron HB-13 exhibited selective hydrolysis of stevioside at β-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 °C. Stevioside determination using the enzymatic assay strongly correlated with results obtained from HPLC quantitation (r2= 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.Mahidol UniversityAgricultural and Biological SciencesChemistryArticleSCOPUS10.1021/acs.jafc.6b05793