Dynamics of biologically active phenolic compounds of Cabernet Sauvignon grape variety during phenophases of maturation, primary processing, vinification and influence on wine antioxidant capacity

Tokom izrade ove doktorske disertacije cilj je bio pratiti dinamiku sadržaja pojedinih fenolnih jedinjenja koja su prisutna u grožđu sorte Cabernet Sauvignon tokom različitih faza sazrevanja, primarne prerade, raznih vidova vinifikacije kao i ispitati antiokisdativni kapacitet novodobijenih vina. Pored vina, značajan deo ove disertacija zauzima istraživanje vezano za kominu koja zaostaje kao nuz proizvod prilikom proizvodnje vina. Ogledi mikrovinifikacije uz primenu različitih kombinacija kvasaca i enzimskih preparata, postavljeni su sa ciljem praćenja dinamike ekstrakcije fenolnih jedinjenja iz čvrstih delova grožđa u vino (3, 5, 7, 14 i 21 dana, respektivno) kao i količine zaostalih fenolnih jedinjenja u komini nakon maceracije. Analizom pripremljenih uzoraka vina identifikovana su sledeća jedinjenja: galna kiselina, protokatehuinska kiselina, p-hidroksibenzoeva kiselina, siringinska, vanilinska, kafeinska, elaginska, p-kumarinska kiselina, katehin, epikatehin i kvercetin. Za svako pojedinačno jedinjenje ispitana je dinamika ekstrakcije, za svaku navedenu varijantu ogleda. Rezultati govore da je p-hidroksibenzoeva kiselina dostigla najraniji maksimum ekstrakcije i to već 3. dan za varijantu ogleda BDX, dok je uz dodati enzimski preparat EXV maksimum dostignut 5. dan maceracije. Među ostalim jedinjenjima najkasniji maksimum i to 21. dan su dostigli katehin, epikatehin, kvercetin, galna i elaginska kiselina za različite kombinacije primene kvasaca i enzimskih preparata. Od svih jedinjenja nađenih u vinu, katehin je imao najviši maksimum ekstrakcije u količini od 36,3108 mg/l u varijanti ogleda BDX. U ovom radu detaljno je analizirana dinamika pojedinih jedinjenja zaostalih u komini, a to su rutin, trans-resveratrol, katehin, epikatehin, vanilin i p-kumarinska kiselina. Katehin i epikatehin su bila najdominantnija jedinjenja nađena u komini grožđa i to onoj koja je macerirala kraći vremenski period pa je zaključeno da je vreme maceracije značajno uticalo na sadržaj pojedinih jedinjenja u komini. Zaključeno je da ova jedinjenja podležu reakcijama desorpcije sve do 14. dana maceracije, nakon čega dolazi do uspostavljanja ravnoteže ili blage desorpcije ili adsorpcije na ćelije kvasca ili komine. Pored navedenih fenolnih jedinjenja, ostala jedinjenja identifikovana u komini su bila siringinska, galna, kafeinska, ferulinska, protokatehuinska, p-hidroksibenzoeva, elaginska, vanilinska kiselina kao i kvercetin, naringenin i kemferol. Izmerena je i antiradikalska i antioksidativna aktivnost uzoraka vina za sve varijante ogleda, gde je nađeno da vreme maceracije značajno utiče na anti-DPPH radikalsku aktivnost vina, ali i to da zasejavanje kljuka različitim kvascima značajno menja antioksidativnost merenu FRAP metodom, u odnosu na antioksidativnost vina spontane fermentacije. Dodatak pojedinih enzimskih preparata tokom maceracije je značajno uticao na antioksidativnu aktivnost vina merenu TEAC testom i FRAP metodom. Pored ogleda dinamike, utvrđen je uticaj rastućeg pritiska ceđenja prevrele komine na povećanje količine pojedinačnih fenolnih jedinjenja u vinu kao i njihovog sadržaja koji je zaostao u komini i komini ceđenoj primenom različitih pritisaka, primenom različitih tehnika vinifikacije kao i tretmana stabilizacije vina. Na osnovu praćenja dinamike fermentacije uz uključivanje i drugih parametara u tehnološki proces proizvodnje, dobijeni su optimalni uslovi proizvodnje vina sa što većim sadržajem biološki aktivnih komponenata značajnih za zdravlje ljudi.content of certain phenolic compounds present in Cabernet Sauvignon grapes during different stages of maturation, primary processing, various types of vinification and to examine the antioxidant capacity of newly produced wines. In addition to wine, a significant part of this dissertation is occupied by research related to the pomace which is present a by-product of wine production. Experiments of microvinification with the use of different combinations of yeasts and enzyme preparations were set to monitor the dynamics of extraction of phenolic compounds from pomace (solid parts of grapes) into wine (3, 5, 7, 14 and 21 days, respectively) as well as the amount of phenolic compounds remaining in fermented pomace after maceration. Through an analysis of prepared wine samples, catechin, epicatechin, quercetin, gallic acid, protocatechuic acid, p-hydroxybenzoic acid, syringic acid, vanillic acid, caffeic acid, ellagic acid and p-coumaric acid were identified. Dynamics extraction was examined for each identified phenolic compound for each mentioned sort of experiment. The results showed that p-hydroxybenzoic acid reached the earliest maximum of extraction on the 3rd day already for the experiment with BDX, while with the added enzyme preparation EXV the maximum was reached on the 5th day of maceration. Among other compounds, catechin, epicatechin, quercetin, gallic and ellagic acid reached the latest maximum on the 21st day for various experiments with different yeast strains and enzyme preparations. Of all the compounds found in wine, catechin had the highest extraction maximum of 36.3108 mg/l during fermentation with BDX yeast. The dynamics of certain compounds such as rutin, trans-resveratrol, catechin, epicatechin, vanillin and p-coumaric acid remained in the pomace were analyzed too. Catechin and epicatechin were the most dominant compounds found in grape pomace, the one that macerated for a shorter period of time, therefore it was concluded that the maceration time significantly affected the content of individual compounds in the pomace. It was concluded that these compounds are subject to desorption reactions until the 14th day of maceration, after which equilibrium is established or mild desorption or adsorption on pomace or yeast cells. In addition to these phenolic compounds, other compounds identified in the pomace were syringic, gallic, caffeic, ferulic, protocatechuic, p-hydroxybenzoic, ellagic, vanillic acid as well as quercetin, naringenin and kaempferol. The antiradical and antioxidant activity of wine samples was measured for all samples, and it was found that the maceration time significantly affects the anti-DPPH radical activity of wine, but also that inoculation with different yeasts significantly changes antioxidant capacity measured by FRAP method, compared to antioxidant capacity of wine obtained by spontaneous fermentation. The addition of enzyme preparations during maceration significantly affected the antioxidant activity of wine measured by TEAC test and FRAP method. Apart from dynamics experiment, the influence that increasing pressing pressure of fermented pomace has on the increase in the amount of individual phenolic compounds as well as on their content remaining in the pomace and pomace drained after applying different pressures, applying various vinification techniques as well as wine stabilization treatments was determined. By monitoring the dynamics of fermentation with the inclusion of other parameters in the technological process of production, optimal conditions for wine production were obtained with the highest possible content of biologically active components important for human health