Article

Separation of Steviol Glycosides by Hydrophilic Liquid Interaction Chromatography

Institut Prof. Dr. Kurz GmbH, Eupener Str. 161, 50933 Cologne, Germany
Food Analytical Methods (Impact Factor: 1.96). 04/2012; 5(2):266-271. DOI: 10.1007/s12161-011-9229-x

ABSTRACT

Stevia rebaudiana Bertoni contains several steviol glycosides with sweet flavour. They all are sweeter than sucrose (up to factor 450). The
various steviol glycosides are difficult to separate by reversed-phase chromatography. In this paper, five different hydrophilic
liquid interaction chromatography columns are characterized using isocratic elution (5–20% water in acetonitrile with buffer
or formic acid). Separation of the steviol glycosides is possible with all but one of the tested columns, but the robustness
of the separation against changes of buffer concentration and percentage of water differ. Aqueous percentage and ion strength
of the eluent are the main factors to be optimized in method development.

Keywords
Stevia rebaudiana
–Hydrophilic liquid interaction chromatography–Steviol glycosides–HPLC–Mass spectrometry

    • "Steviol glycosides were quantified by HPLC as published elsewhere (Morlock et al., 2014; Zimmermann et al., 2011). Briefly, steviol glycosides were analyzed by HPLC-UV using a Hilic column (Nucleodur Hilic, 125 mm  2.1 mm, 3 mm particle size; Macherey-Nagel, Düren, Germany) with a gradient elution and acetonitrile and water as eluents. "
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    ABSTRACT: The aim of our study was to determine the antioxidant activities, cytotoxicity and proliferative properties in Stevia rebaudiana leaves and stems. Leaves extracts exhibited a higher antioxidant activity than stems extract, through oxygen radical absorbance capacity (ORAC) and cellular antioxidant activity (CAA) assays. Stevioside and rebaudioside A, the main sweetening metabolites in stevia leaves, exhibited a low ORAC value in comparison with plant extracts, while did not elicit any CAA. Stevia rebaudiana did not exhibit toxicity against HepG2 (hepatocellular carcinoma) human cells. No proliferative nor catalase modulations were observed in cells treated with such extracts. Our findings support the promising role of stevia that, apart from its sweetness, can act as a source of antioxidants, even at the intracellular level. This activity makes S. rebaudiana crude extract an interesting resource of natural sweetness with antioxidant properties which may find numerous applications in foods and nutritional supplements industries.
    No preview · Article · May 2015 · International Journal of Food Sciences and Nutrition
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    • "[33] [34] The growing interest in the use of steviol glycosides has motivated research aimed to characterize and separate these sweetening agents. [35] [36] [37] [38] For separation, high-performance liquid chromatography (HPLC) using amino, [39] [40] [41] C18 [42] [43] and HILIC columns [44] [45] is considered the method of choice. Detection has been performed by optical spectroscopy [46] and MS, [47] [48] [49] [50] including direct approaches using desorption/ionization ambient MS techniques. "
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    ABSTRACT: Traveling wave ion mobility mass spectrometry (TWIM-MS) is shown to be able to separate and characterize several isomeric forms of diterpene glycosides stevioside (Stv) and rebaudioside A (RebA) that are cationized by Na(+) and K(+) at different sites. Determination and characterization of these coexisting isomeric species, herein termed catiomers, arising from cationization at different and highly competitive coordinating sites, is particularly challenging for glycosides. To achieve this goal, the advantage of using CO2 as a more massive and polarizable drift gas, over N2 , was demonstrated. Post-TWIM-MS/MS experiments were used to confirm the separation. Optimization of the possible geometries and cross-sectional calculations for mobility peak assignments were also performed. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.
    Full-text · Article · Feb 2015 · Journal of Mass Spectrometry
    • "Steviol glycosides were quantified by HPLC as published elsewhere (Morlock et al., 2014; Zimmermann et al., 2011). Briefly, steviol glycosides were analyzed by HPLC-UV using a Hilic column (Nucleodur Hilic, 125 mm  2.1 mm, 3 mm particle size; Macherey-Nagel, Düren, Germany) with a gradient elution and acetonitrile and water as eluents. "

    No preview · Chapter · Jan 2015
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