Zwitterionic hydrophilic interaction chromatography coupled with post-column derivatization for the analysis of glutathione in wine samples |
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Authors: | Constantinos K. Zacharis Paraskevas D. Tzanavaras Theano D. Karakosta Demetrius G. Themelis |
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Affiliation: | 1. Laboratory of Analytical Chemistry, Department of Chemistry, Aristotle University of Thessaloniki, GR-54124 Thessaloniki, Greece;2. Research Laboratory for the Physical and Chemical Testing of Foods, Department of Food Technology, School of Food Technology and Nutrition, Alexander Technological Educational Institute (ATEI) of Thessaloniki, 57400 Thessaloniki, Greece |
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Abstract: | In this study the development, validation and application of a new chromatographic method for the determination of glutathione (GSH) in wine samples is presented. The separation of the GSH was carried out using a sulfobetaine-based hydrophilic interaction chromatography (HILIC) analytical column whereas its detection was carried out spectrofluorimetrically (λext/λem = 340/455 nm) after post-column derivatization with o-phthalaldehyde. GSH was separated efficiently from matrix endogenous compounds of wines by using a mobile phase of 15 mmol L−1 CH3COONH4 (pH = 2.5)/CH3CN, 35/65% (v/v). The parameters of the post-column reaction (pH, amount concentration of the reagent and buffer solution, flow rate, length of the reaction coil) were investigated. The linear determination range for GSH was 0.25–5.0 μmol L−1 and the LOD was 19 nmol L−1. No matrix effect was observed, while the accuracy was evaluated with recovery experiments and was ranged between 89% and 108%. |
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Keywords: | Glutathione Post-column derivatization Hydrophilic interaction chromatography Wines o-Phthalaldehyde |
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