Title: Steviol Quantification at the picomol level by HPLC
Authors: Minne, Veerle
Compernolle, Frans
Toppet, Suzanne
Geuns, Jan # ×
Issue Date: Jan-2004
Publisher: American Chemical Society, Books and Journals Division
Series Title: Journal of Agricultural and Food Chemistry vol:52 issue:9 pages:2445-2449
Abstract: A simple and highly sensitive reversed-phase high-performance liquid chromatographic method (RP-HPLC) has been developed for the determination of steviol (SV) using dihydroisosteviol (DHISV) as an internal standard (IS). SV and DHISV were derivatized by reaction of the acids with 4-(bromomethyl)-7-methoxycoumarin in an aprotic solvent (DMF or acetone). The resulting ester derivatives were separated on an ODS column (250 x 4.6 mm i.d., 5 mum particle size) using fluorescence detection with excitation at 321 nm and emission at 391 nm. The mobile phase consisted of acetonitrile/water (80:20 v/v) with a flow rate of 1 mL min(-1). A linear relationship was observed for concentrations between 0.5 and 50 mug/mL of SV, and the detection limit was 100 pg. For application of this method to samples of beer fortified with stevioside, a simple procedure for extraction of the beer with diethyl ether and derivatization in DMF was applied. Whereas beer samples spiked with SV gave a linear response over the range 0.1-15 mug/mL beer, no SV could be detected in beer samples enriched in stevioside that had been stored for over 3 years. The application of the method to plant samples involved preparation of an acid fraction containing the SV analyte, derivatization, and sample cleanup using small silica columns and thin-layer chromatography. A sensitive determination of 594 ng of steviol present in 100 mg of dry plant material was performed with high precision and accuracy.
ISSN: 0021-8561
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Molecular Design and Synthesis
Molecular Physiology of Plants and Micro-organisms Section - miscellaneous
× corresponding author
# (joint) last author

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