Edoardo Longo

Isotopic Exchange HPLC-HRMS/MS Applied to Cyclic Proanthocyanidins in Wine and Cranberries

AbstractCyclic B-type proanthocyanidins in red wines and grapes have been discovered recently. However, proanthocyanidins of a different chemical structure (non-cyclic A-type proanthocyanidins) already known to be present in cranberries and wine possess an identical theoretical mass. As a matter of fact, the retention times and the MS/MS fragmentations found for the proposed novel cyclic B-type tetrameric proanthocyanidin in red wine and the known tetrameric proanthocyanidin in a cranberry extract are herein shown to be identical. Thus, hydrogen/deuterium (H/D) exchange was applied to HPLC-HRMS/MS to confirm the actual chemical structure of the new oligomeric proanthocyanidins. The comparison of the results in water and deuterium oxide and between wine and cranberry extract indicates that the cyclic B-type tetrameric proanthocyanidin is the actual constituent of the recently proposed novel tetrameric species ([C60H49O24]+, m/z 1153.2608). Surprisingly, the same compound was also identified as the main tetrameric proanthocyanidin in cranberries. Finally, a totally new cyclic B-type hexameric proanthocyanidin ([C90H73O36]+, m/z 1729.3876) belonging to this novel class was identified for the first time in red wine. Graphical Abstractᅟ

Monitoring of Glucose in Beer Brewing by a Carbon Nanotubes Based Nylon Nanofibrous Biosensor

This work presents the design, preparation, and characterization of a novel glucose electrochemical biosensor based on the immobilization of glucose oxidase (GOX) into a nylon nanofibrous membrane (NFM) prepared by electrospinning and functionalized with multiwalled carbon nanotubes (CNT). A disc of such GOX/CNT/NFM membrane (40 źm in thickness) was used for coating the surface of a glassy carbon electrode. The resulting biosensor was characterized by cyclic voltammetry and chronoamperometry, with ferrocene methanol as mediator. The binding of GOX around the CNT/NFM greatly enhances the electron transfer, which results in a biosensor with a current five times higher than without CNT. The potential usefulness of the proposed biosensor was demonstrated with the analysis of glucose in commercial beverages and along the monitoring of the brewing process for making beer, from the mashing to the fermentation steps.

Selective binding of potassium and calcium ions to novel cyclic proanthocyanidins in wine by high-performance liquid chromatography/high-resolution mass spectrometry

RATIONALE Cyclic B-type (referred also as crown) proanthocyanidins were recently identified in wines. An HPLC-HRMS/MS method was applied to study the binding of cyclic and non-cyclic PAC to potassium and calcium ions, which affect the chemico-physical stability of wines. Different binding affinities suggest that cyclic and non-cyclic analogues, despite the equal number of monomer units, influence the colloidal stability of wine and are related to the grape variety or winemaking conditions. METHODS Nineteen red and white wines were analysed by HPLC high resolution tandem mass spectrometry with positive electrospray ionization to investigate the distribution of novel cyclic proanthocyanidins and their calcium, potassium and sodium adducts. Principal Component Analysis was used to study the distribution of the wines and the relationships among proanthocyanidins with and without cation complexes. RESULTS A dependence on specific isomers (and conformations) was found for the non-cyclic procyanidin (PC) trimer whereas the cyclic tetrameric PAC were shown to bind better to potassium than their non-cyclic analogues. The binding to these metals appeared to be influenced not only be the number of monomer units but also by the conformation assumed by the molecules. Statistical analysis evidenced that the adducts distribution in different wines is less influenced by the grape variety used for winemaking than their associated [M+H]+ species studied earlier. CONCLUSIONS Wines from 19 grape varieties were investigated in order to identify potassium and calcium complexes of non- and cyclic B-type proanthocyanidins that were recently discovered. The results showed a dependence of the distribution of metal complexes according to the cyclic or non cyclic geometry of proanthocyanidins. The multivariate analysis of the mass spectrometric results showed a relationship with the grape variety, however not so straightforward as evidenced for the non-complexed species.

High resolution mass approach to characterize refrigerated black truffles stored under different storage atmospheres

Freshly harvested Tuber melanosporum samples were packed and stored at 4°C under reduced atmospheric pressure or modified atmosphere for four weeks. Multivariate analysis was employed to correlate the antioxidant power of the ethanolic extracts of the samples with the chemical composition determined by high resolution mass spectrometry. High performance liquid chromatography coupled with a coularray detector was applied to select the chemical species associated with the antioxidant power. Four classes of chemical compounds were investigated in more detail by a targeted approach: derivatives of glutathione, adenine (such as S-adenosyl-homocysteine), oxidized linoleic acid and ergosterol. Adducts containing glutathione and adenine with oxidized linoleic acid were observed in TM for the first time and can be considered markers of freshness of the product. S-adenosyl-homocysteine, the acetyl-carnitine adduct with cysteinyl-glycine and several oxidized linoleic acid derivatives were among the markers of degradation.

Effect of light irradiation on the antioxidant stability of oleuropein

The stability of oleuropein, a natural antioxidant from Olea europaea, has been often studied in connection with thermal or enzymatic treatments, but very little is known about the effects of UV light. This work aimed at studying the UV-C effects on oleuropein standard solutions once dissolved in ethanol or water. During irradiation, aliquots were taken and analyzed by a flow injection system equipped with a multi-channel coulometric detector and a high-resolution mass spectrometer. The effects of irradiation were also studied by UV spectroscopy. The results show that oleuropein is relatively stable in water or ethanol, but that under UV-C light undergoes a series of fast decomposition reactions leading to hydroxytyrosol and elenolic acid. Overall, this study provides evidences that the degradation of oleuropein by UV-C light follows a mechanism dependent on the solvent used. Moreover, the solvent affects the resulting redox properties of the solution.

Disambiguation of Isomeric Procyanidins with Cyclic B-Type and Non-cyclic A-Type Structures from Wine and Peanut Skin with HPLC-HDX-HRMS/MS

AbstractHydrogen/deuterium exchange coupled with high-resolution mass spectrometry was successfully applied for the identification of A-type tetrameric, pentameric, and hexameric procyanidins in peanut skin. This extended a previous study on isomeric cyclic B-type unconventional tetramer, pentamer, and hexamer procyanidins found in wine and cranberries. Not only had the method successfully identified the procyanidins with a single A-linkage (e.g., tetrameric m/z 1153.2608) by means of distinguishing them from their isomeric cyclic B-type analogues, but this method also worked for procyanidins with two or more A-linkages (such as the tetrameric m/z 1151.2452). As a further consequence, B-type cyclic pentamers and hexamers in wine have been elucidated with hydrogen/deuterium exchange (HDX) for the first time. Graphical Abstractᅟ

Relative abundances of novel cyclic prodelphinidins in wine depending on the grape variety

The identification of cyclic B-type procyanidins in grape and wine was recently disclosed. Some of these were also found in berries of totally different vegetal species (eg, Vaccinium sp.). However, presence of a wider class of these cyclic condensed tannin compounds with variably substituted monomers has never been addressed so far. Here, an extended list of oligomeric cyclic proanthocyanidins (PAC) bearing variable substitution patterns on the main flavan-3-ol unit has been searched in wines. Nearly 7600 theoretical structures were calculated and searched in red and white wine samples made from different grape varieties. Moreover, a hydrogen/deuterium exchange approach (already applied to a cyclic tetrameric procyanidin) coupled to high-resolution mass spectrometry was applied to confirm their cyclic B-type structure rather than a non cyclic A-type structure, otherwise isomeric and undistinguishable by LC-MS alone. The main group of novel cyclic PAC observed is shown to contain (epi)gallocatechin beside (epi)catechin as the constituent monomers.