Dimitrios Tsimogiannis

Characterization of Flavonoid Subgroups and Hydroxy Substitution by HPLC-MS/MS

HPLC-DAD coupled with mass spectrometry in the positive ionization mode was applied to study the fragmentation of twelve selected flavonoids. Compounds belonging to all the major subgroups found in common plants, i.e. flavonols, flavones, dihydroflavonols, flavanones and flavanols were studied. Compound standards were injected into the spectrometer and produced characteristic mass spectra. The fragmentation of each compound was studied and it was shown that the dehydration and carbon monoxide losses from the [M+H]+ ion by the members of each subgroup produced specific fragments, thus allowing the characterization of the flavonoid subgroups. Moreover, fragments resulting from fission of the C-rings are specific of each subgroup and revealed the substitution pattern of A- and B-rings. In order to verify the identifying efficiency of the positive ionization mode through these characteristic fragmentations, the unknown flavonoids of an Origanum vulgare diethyl ether extract were separated with the HPLC system and the major peaks were successfully identified with the mass spectrometer.

Anthocyanin copigmentation and color of wine: The effect of naturally obtained hydroxycinnamic acids as cofactors.

Copigmentation of anthocyanins accounts for over 30% of fresh red wine color, while during storage, the color of polymeric pigments formed between anthocyanins and proanthocyanidins predominates. Rosmarinic acid and natural extracts rich in hydroxycinnamic acids, obtained from aromatic plants (Origanum vulgare and Satureja thymbra), were examined as cofactors to fresh Merlot wine and the effect on anthocyanin copigmentation and wine color was studied during storage for 6months. An increase of the copigmented anthocyanins that enhanced color intensity by 15-50% was observed, confirming the ability of complex hydroxycinnamates to form copigments. The samples with added cofactors retained higher percentages of copigmented anthocyanins and higher color intensity, compared to the control wine, up to 3 months. However, the change in the equilibrium between monomeric and copigmented anthocyanins that was induced by added cofactors, did not affect the rate of polymerization reactions during storage.

Evolution of phenolic compounds and metal content of wine during alcoholic fermentation and storage.

Changes in the principal phenolic compounds and metal content during the vinification process and storage under modified atmosphere (50% N2, 50% CO2) of Merlot and Syrah wines, from grapes cultivated in Greece, have been investigated. Comparing the variation of metals at maceration process, with the variation of monomeric anthocyanins and flavonols, an inverse relationship was noticed, that can be attributed to complexing reactions of polyphenols with particular trace elements. Cu decreased rapidly, whereas a similar behavior that could be expected for Fe and Mn was not confirmed. Differences in the profile of anthocyanins and flavonols in the fresh Merlot and Syrah wines are reported. During 1 year of storage monomeric anthocyanins declined almost tenfold, probably due to polymerization reactions and copigmentation. Also, a decrease in flavonol glycosides and increase in the respective aglycones was observed, attributed to enzymatic hydrolysis. The concentration of total phenols and all metals remained practically constant.

Characteristic phenolic composition of the Greek variety Mavrokountoura grape and wine

Mavrokountoura, a clone of the Mandilaria grape cultivar, was studied for the first time. The morphological characteristics and composition of berries were determined, as well as total phenols and tannins in skins, flesh, and seeds. The skin anthocyanins were identified using HPLC-MS/MS and quantified using HPLC-DAD. Two main anthocyanins were identified as malvidin-3-O-glucoside and malvidin-3-O-(6″-p-coumaroyl)-glucoside. One minor anthocyanin was identified as malvidin-3-O-(6″-acetyl)-glucoside. Glycosides of delphinidin, petunidin, and peonidin were detected in traces. Young Mavrokountoura wine was also studied. Extraction of total phenols from the grape to the young wine amounted to 32%, composing mainly non-tannic phenols. The individual anthocyanin components exhibited different extractability values from the skin to wine, ranging between 10–94%.