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Identification and Quantification of Major Steviol Glycosides in Stevia rebaudiana Purified Extracts by 1H NMR Spectroscopy

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    Identification and Quantification of Major Steviol Glycosides in Stevia rebaudiana Purified Extracts by 1H NMR Spectroscopy
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    † Institute of Pharmacy/Pharmacognosy, Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, Innrain 52c, 6020 Innsbruck, Austria
    ‡ Prodalysa Ltda, Lote 13, Parque Industrial Gulmue, Camino Internacional, Concón, Chile
    *Tel: +43-512-507-5300. Fax: +43-512-507-2939. E-mail: [email protected]
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    Journal of Agricultural and Food Chemistry

    Cite this: J. Agric. Food Chem. 2011, 59, 9, 4378–4384
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    https://doi.org/10.1021/jf104922q
    Published March 21, 2011
    Copyright © 2011 American Chemical Society
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    Abstract

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    The use of 1H NMR spectroscopy for the characterization of Stevia rebaudiana extracts is presented. The developed method allows qualitative and quantitative determination of the major steviol glycosides in purified extracts and fractions obtained from various stages of the purification process. Moreover, it proved to be a powerful tool to differentiate between glycosides which are naturally occurring in the stevia plant and artifacts formed in the course of the manufacturing process. Identification of steviol glycosides was achieved by the use of 2D NMR techniques, whereas quantification is based on qHNMR using anthracene as internal standard. The solvent mixture pyridine-d5−DMSO-d6 (6:1) enabled satisfactory separation of the signals to be integrated. Validation of the method was performed in terms of specificity, precision, accuracy, linearity, robustness, and stability. Quantitative results were compared to those obtained with the JECFA HPLC−UV method and were found to be in reasonable agreement. NMR analysis does not rely on the use of reference compounds and enables significantly faster analysis compared to HPLC−UV. Thus, NMR represents a feasible alternative to HPLC-based methods for the quality control of Stevia rebaudiana extracts.

    Copyright © 2011 American Chemical Society

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    Selected 1D and 2D NMR data; HPLC−MS and HPLC−UV chromatograms; analysis of the minor steviol glycosides by HPLC−UV. This material is available free of charge via the Internet at http://pubs.acs.org.

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    Journal of Agricultural and Food Chemistry

    Cite this: J. Agric. Food Chem. 2011, 59, 9, 4378–4384
    Click to copy citationCitation copied!
    https://doi.org/10.1021/jf104922q
    Published March 21, 2011
    Copyright © 2011 American Chemical Society
    Request reuse permissions

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