The wine sector is an important part of the international agrifood industry, as shown by the OIV report on 2022 world wine production, estimated between 257.5 and 262.3 millions of hL. In parallel with the winemaking process a considerable amount of by-products are produced, with a potential negative impact on the environment. In this regard, the OIV in the 2020-2024 strategic plan encourages the circular economy through the reuse of waste and the management of by-products, defining the guidelines of the green economy. Grape skins and seeds are among the most abundant and interesting wine making by-products due to their content in phenolic compounds (e.g., flavonoids), with potential health-promoting effects. Among them, the antioxidant activity (AOA) is one of most investigated properties since oxidative stress is a key factor in the induction of chronic diseases and the relative progression. There are many assays developed for the in vitro evaluation of AOA; in order to obtain a complete evaluation, it is necessary to apply in parallel different methods. In this study, AOA of grape by-products was measured spectrophotometrically, using 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH) and ferric reducing antioxidant power (FRAP) assays. Several studies have shown that oxidative stress and type 2 diabetes (T2D) are closely correlated. In modern society, T2D is one of the main public health problems and for its management, various approaches are used, such as: the suppression of glucose absorption by inhibiting the enzymes that hydrolyze carbohydrates (α-amylase, α-glucosidase), or inhibition of dipeptidyl peptidase-IV (DPP-IV) activity. Grapes and derivatives have shown promising results in modulating post-prandial glycaemia; however, few data have been produced to investigate their impact on α-amylase and dipeptidyl peptidase-IV (DPP-IV). After a preliminary characterization of the AOA, the most promising matrices were selected. The DPP-IV inhibitory activity test was applied to these matrices, performed using the DPP-IV inhibitor screening kit (MAK203): 50 μL of enzyme solution and 25 μL of sample solution were premixed and incubated at 37°C for 10 min; then 25 μL of DPP-IV substrate was added. Fluorescence (λex=360 nm, λem=460 nm) was measured once every min up to 60 min. The efficacy of the test inhibitors was compared to the DPP4 inhibitor (sitagliptin) provided as a control, and to remove background interference, the buffer was added instead of enzyme solution. Each sample was tested in triplicate and relative DPP-IV inhibition (as percentage) was calculated. Statistical analyzes were performed using the IBM SPSS Statistics, Version 27.0. The results showed that winemaking by-products contain antioxidant phenolic compounds, supporting their use as a source of healthy compounds. Furthermore, the preliminary results obtained by DPP-IV inhibitory activity assay show that the selected samples were active in modulating dipeptidyl peptidase-IV, with a potential positive effect on glycaemia. On this basis, grape pomace could be used as an ingredient in the formulation of healthy products, implementing, in parallel, the circular economy.
Il settore vitivinicolo occupa un ruolo importante nell'industria agroalimentare internazionale: nel 2022, la produzione mondiale di vino è stata stimata tra 257,5 e 262,3 milioni di hL. Parallelamente al processo di vinificazione si producono notevoli quantità di sottoprodotti di lavorazione, i quali hanno un importante impatto negativo sull'ambiente. A questo proposito, l'OIV nel piano strategico 2020-2024 incentiva l'economia circolare attraverso il riutilizzo dei rifiuti e la gestione dei sottoprodotti enologici, definendo le linee guida della green economy. Le bucce e i semi dell'uva sono tra i sottoprodotti enologici più abbondanti e interessanti grazie al loro contenuto di composti fenolici (es. flavonoidi), con potenziali effetti benefici per la salute. Tra queste, l'attività antiossidante (AOA) è una delle proprietà più studiate poiché lo stress ossidativo è un fattore chiave nell'induzione di malattie croniche e nella loro relativa progressione. Esistono molti saggi in vitro sviluppati per la valutazione dell'AOA; tuttavia, al fine di ottenere una valutazione completa è necessario applicare in parallelo differenti metodiche. In questo studio, l'AOA dei sottoprodotti dell'uva è stata misurata spettrofotometricamente, utilizzando i saggi del radicale libero 1,1-difenil-2-picryl-idrazil (DPPH) e del potere antiossidante di riduzione del ferro (FRAP). Diversi studi hanno dimostrato che lo stress ossidativo e il diabete di tipo 2 (T2D) sono strettamente correlati. Nella società moderna, il T2D è uno dei principali problemi di salute pubblica e per la sua gestione vengono utilizzati vari approcci, come: la soppressione dell'assorbimento del glucosio mediante l'inibizione degli enzimi che idrolizzano i carboidrati (α-amilasi, α- glucosidasi) o l'inibizione dell'attività della dipeptidil peptidasi-IV (DPP-IV). Uve e derivati hanno mostrato risultati promettenti nella modulazione della glicemia postprandiale; tuttavia, sono stati prodotti pochi dati per studiare il loro impatto sull'α-amilasi e sulla dipeptidil peptidasi-IV (DPP-IV). Dopo una caratterizzazione preliminare dell'AOA, sono state selezionate le matrici più promettenti. A tali matrici è stato applicato il test di attività inibitoria della DPP-IV, eseguito utilizzando il DPP-IV inhibitor screening kit (MAK203): 50 μL di soluzione enzimatica e 25 μL di soluzione campione sono stati premiscelati e incubati a 37°C per 10 min; quindi, sono stati aggiunti 25 μL di substrato DPP-IV. La fluorescenza (λex=360 nm, λem=460 nm) è stata misurata una volta ogni min fino a 60 min. L'efficacia degli inibitori del test è stata confrontata con l'inibitore DPP4 (sitagliptin) fornito come controllo e, per rimuovere l'interferenza di fondo, è stata aggiunta la soluzione tampone invece della soluzione enzimatica. Ciascun campione è stato saggiato in triplicato ed è stata calcolata l'inibizione relativa (in percentuale). Le analisi statistiche sono state eseguite utilizzando IBM SPSS Statistics, versione 27.0. I risultati hanno mostrato che i sottoprodotti della vinificazione contengono composti fenolici antiossidanti, il che consente di ipotizzare un loro utilizzo come fonte di composti salutistici. Inoltre, i risultati preliminari ottenuti dal saggio di attività inibitoria della DPP-IV hanno mostrato che i campioni selezionati erano attivi nella modulazione della dipeptidil peptidasi-IV, con un potenziale effetto positivo sulla glicemia. Su questa base, le vinacce potrebbero essere utilizzate come ingredienti nella formulazione di prodotti salutistici, promuovendo, parallelamente, l'economia circolare.
In vitro methods to evaluate the potential health-promoting effects of winemaking by-products / C. Bani, P.A. Restani, F. Mercogliano, C.M. DI LORENZO - In: Vitiviniculture and Information Technologies[s.l] : OIV, 2023 Jun. - ISBN 978-2-85038-083-9. - pp. 825-825 (( Intervento presentato al 44. convegno World congress of vine and wine tenutosi a Cádiz /Jerez nel 2023.
In vitro methods to evaluate the potential health-promoting effects of winemaking by-products
C. BaniPrimo
;P.A. Restani;F. Mercogliano;C.M. DI LORENZOUltimo
2023
Abstract
The wine sector is an important part of the international agrifood industry, as shown by the OIV report on 2022 world wine production, estimated between 257.5 and 262.3 millions of hL. In parallel with the winemaking process a considerable amount of by-products are produced, with a potential negative impact on the environment. In this regard, the OIV in the 2020-2024 strategic plan encourages the circular economy through the reuse of waste and the management of by-products, defining the guidelines of the green economy. Grape skins and seeds are among the most abundant and interesting wine making by-products due to their content in phenolic compounds (e.g., flavonoids), with potential health-promoting effects. Among them, the antioxidant activity (AOA) is one of most investigated properties since oxidative stress is a key factor in the induction of chronic diseases and the relative progression. There are many assays developed for the in vitro evaluation of AOA; in order to obtain a complete evaluation, it is necessary to apply in parallel different methods. In this study, AOA of grape by-products was measured spectrophotometrically, using 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH) and ferric reducing antioxidant power (FRAP) assays. Several studies have shown that oxidative stress and type 2 diabetes (T2D) are closely correlated. In modern society, T2D is one of the main public health problems and for its management, various approaches are used, such as: the suppression of glucose absorption by inhibiting the enzymes that hydrolyze carbohydrates (α-amylase, α-glucosidase), or inhibition of dipeptidyl peptidase-IV (DPP-IV) activity. Grapes and derivatives have shown promising results in modulating post-prandial glycaemia; however, few data have been produced to investigate their impact on α-amylase and dipeptidyl peptidase-IV (DPP-IV). After a preliminary characterization of the AOA, the most promising matrices were selected. The DPP-IV inhibitory activity test was applied to these matrices, performed using the DPP-IV inhibitor screening kit (MAK203): 50 μL of enzyme solution and 25 μL of sample solution were premixed and incubated at 37°C for 10 min; then 25 μL of DPP-IV substrate was added. Fluorescence (λex=360 nm, λem=460 nm) was measured once every min up to 60 min. The efficacy of the test inhibitors was compared to the DPP4 inhibitor (sitagliptin) provided as a control, and to remove background interference, the buffer was added instead of enzyme solution. Each sample was tested in triplicate and relative DPP-IV inhibition (as percentage) was calculated. Statistical analyzes were performed using the IBM SPSS Statistics, Version 27.0. The results showed that winemaking by-products contain antioxidant phenolic compounds, supporting their use as a source of healthy compounds. Furthermore, the preliminary results obtained by DPP-IV inhibitory activity assay show that the selected samples were active in modulating dipeptidyl peptidase-IV, with a potential positive effect on glycaemia. On this basis, grape pomace could be used as an ingredient in the formulation of healthy products, implementing, in parallel, the circular economy.File | Dimensione | Formato | |
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