Abstract
In this work, high performance liquid chromatography (HPLC) and fluorescence spectrometry methods for determination of natural coumarins in Tokaj wine were developed and compared. Molecularly imprinted solid phase extraction procedure was applied for sample preparation. The separation of esculin, coumarin, herniarin, 4-methylumbelliferone, scoparone, scopoletin was performed on core-shell C18 type of stationary phase (100 × 4.6 mm, 5 µm) with a gradient elution of mobile phase containing 1% aqueous solution of acetic acid and methanol, UV-VIS (280 nm for coumarin) and fluorescence detection (Ex 320 nm, Em 450 nm for other coumarins). The HPLC method was validated in term of linearity, limit of detection, limit of quantification, precision and accuracy. Fluorescence detection offers high sensitivity with limit of detection in the ng mL-1 range. Scopoletine and 4-methylumbelliferone were identified and quantified in tested wines. Emission spectral data, synchronous fluorescence spectra of coumarins from fluorescence spectrometry and total concentrations of coumarins quantified by the HPLC method were used in the partial least squares regression. The linear regression between the concentrations predicted by the partial least squares model versus true values obtained by HPLC method confirmed good agreement between the two methods.