Advanced liquid chromatography/mass spectrometry profiling of anthocyanins in relation to set of red wine varieties certified in Czech Republic.

Hyphenation of micro-liquid and ultra-performance liquid chromatography with a hybrid (QqTOF) tandem mass spectrometry was studied from the viewpoint of their applicability for monitoring of anthocyanin dyes in various cultivars and different vintages of red wine. After appropriate optimization both techniques proved to be suitable for this task. UHPLC system provided lower LOD and LOQ values as well as higher productivity and precision of retention parameters and peak areas with respect to micro-LC method. On the other hand, micro-LC method offers significant savings of mobile phase, less contamination of ion source of mass spectrometer and makes the nebulization process in electrospray much easier. In combination with principal component analysis (PCA), the method allows to elucidate long-term relations in a complete set of certified red wine cultivars grown in South Moravia (certified in Czech Republic until year 2005). Among the studied varieties Rubinet exhibit a specific anthocyanin profile. Especially interesting is a high content of 3-coumaroylglucoside-5-glucosides of malvidin and peonidin. Those dyes can be used as markers of artificial color enhancement. Principal component analysis applied to data processed with regard to enzymatic activity markedly facilitate classification among varieties. This approach allows resolution of teinturiers, variety Blaufränkish and partial classification of other varieties.

Analysis of anthocyanin pigments in Lonicera (Caerulea) extracts using chromatographic fractionation followed by microcolumn liquid chromatography-mass spectrometry.

Anthocyanins from the fruit Lonicera caerulea L. var. kamtschatica (blueberry honeysuckle, Caprifoliaceae) were studied via (semi)preparative chromatographic fractionation followed by MS and µLC/MS analysis. The extraction procedure was optimized with respect to analytical purposes as well as its potential use for the preparation of nutraceuticals. The highest yield of anthocyanins was obtained using acidified methanol as the extraction medium. A comparable total anthocyanin content was obtained using a mixture of methanol and acetone. However, when Lonicera anthocyanins were in contact with acetone, a condensation reaction occurred to a large extent and related 5-methylpyranoanthocyanins were found. The effect of other extraction media, including ethanol as a "green" solvent, is also discussed. The potential of two fractionation procedures for extract purification differing in their chromatographic selectivity and scale was studied (i.e. using a Sephadex LH-20 gel column and a reversed phase). Fractions obtained by both procedures were used for a detailed analysis. MS and µLC/MS(2) methods were used for monitoring anthocyanin and 5-methylpyranoderivatives content as well as identifying less common and more complex dyes (dimer of cyanidin-3-hexoside, cyanidin-ethyl-catechin-hexosides, etc.). These more complex dyes are most likely formed during fruit treatment.