Publication: Development and Validation of an Enzymatic Method to Determine Stevioside Content from Stevia rebaudiana
Issued Date
2017-04-19
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ISSN
15205118
00218561
00218561
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2-s2.0-85017654501
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of Agricultural and Food Chemistry. Vol.65, No.15 (2017), 3223-3229
Suggested Citation
Somsiri Udompaisarn, Dumrongkiet Arthan, Jamorn Somana Development and Validation of an Enzymatic Method to Determine Stevioside Content from Stevia rebaudiana. Journal of Agricultural and Food Chemistry. Vol.65, No.15 (2017), 3223-3229. doi:10.1021/acs.jafc.6b05793 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/41571
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Title
Development and Validation of an Enzymatic Method to Determine Stevioside Content from Stevia rebaudiana
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Abstract
© 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.