Diferencijacija različitih stilova vina Malvazija istarska na osnovi aromatskog profila

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1 IZVORNI ZNANSTVENI RAD Diferencijacija različitih stilova vina Malvazija istarska na osnovi aromatskog profila Igor LUKIĆ 1, Sanja RADEKA 1, Andreja VANZO 2, Klemen LISJAK 2, Dejan BAVČAR 2 1 Institut za poljoprivredu i turizam, Karla Huguesa 8, Poreč, Hrvatska, ( 2 Kmetijski inštitut Slovenije, Hacquetova ulica 17, 1000 Ljubljana, Slovenija Sažetak U svrhu diferencijacije različitih stilova vina od sorte Malvazija istarska na osnovi aromatskog profila, prikupljen je 41 uzorak i podvrgn-ut GC/MS analizi. Dobiveni podaci obrađeni su kemometrijski. Svježa vina Malvazije sadržavala su više koncentracije većine monoterpena i β-damascenona, kao i estera i kiselina. Desertna vina bila su obilježena povišenim udjelima monoterpenola i etil-acetata. Vina iz barrique bačava, te ona proizvedena tradicionalnom tehnologijom, sadržavala su više koncentracije norizoprenoida, estera tercijarne arome, hlapljivih fenola, laktona i furana. Tradicionalna vina Malvazije odlikovala su se povišenim koncentracijama metanola i viših alkohola. Ključne riječi: Malvazija istarska, stil vina, aromatski profil Differentiation of different Malvazija istarska wine styles based on wine aromatic profiles Abstract With the purpose of differentiation of Malvazija istarska wine styles based on wine aromatic profiles, 41 samples were collected and subjected to GC/MS analysis. Obtained data were analysed using chemometrics. Fresh Malvazija wines contained higher concentrations of monoterpenes, β-damascenone, esters and acids. Dessert wines were marked by high proportions of monoterpenols and ethyl acetate. Wines from barrique barrels, and those produced by traditional technology, contained higher concentrations of norisoprenoids, tertiary aroma esters, volatile phenols, lactones, and furans. Traditional Malvazija wines were characterized by elevated concentrations of methanol and higher alcohols. Key words: Malvazija istarska, wine style, aromatic profile Proceedings. 50 th Croatian and 10 th International Symposium on Agriculture. Opatija. Croatia ( ) Section 8. Viticulture and Enology 515

2 Igor LUKIĆ, Sanja RADEKA, Andreja VANZO, Klemen LISJAK, Dejan BAVČAR Uvod Malvazija Istarska (Vitis vinifera L.) domaća je i vodeća sorta bijelog grožđa u hrvatskoj Istri, druga po zastupljenosti u Republici Hrvatskoj, dok je u slovenskom dijelu Istre također od izuzetne važnosti. U posljednje vrijeme dodatno je iskorištena i djelomično valorizirana njezina prilagođenost i mogućnost prerade širokim rasponom različitih tehnoloških rješenja uz dobivanje visokokvalitetnih vina, pa se tako na tržištu mogu pronaći brojni stilovi. Međutim, raznolikost stilova vina Malvazije koji još nisu jasno i precizno definirani i klasificirani na neki način zbunjuje većinu potrošača, te otežava prepoznavanje stila koji najbolje odgovara njihovim specifičnim željama. Isto tako, nedefinirani i nediferencirani stilovi, kao i nepoznavanje njihovih ključnih karakteristika, onemogućuju potpuno iskorištenje njihovih specifičnih potencijala i ciljano upravljanje proizvodnjom u svrhu postizanja bolje kvalitete. Iz tog razloga, potrebna je dodatna, temeljita stilska karakterizacija i diferencijacija na znanstvenim osnovama. Iako postoje objavljeni podaci o karakteristikama vina Malvazije (Bavčar i sur., 2011; Lukić i sur., 2008; Radeka i sur., 2008), oni su nedovoljni i nepotpuni da posluže kao temelj za ponovnu procjenu i definiciju. Kako su hlapljivi spojevi arome jedan od stupova senzorne kvalitete i prepoznatljivosti stila vina, u ovom je radu izučavan aromatski profil četiri prethodno identificirana glavna stila vina Malvazije istarske, i to u svrhu njihove diferencijacije. U studiju su uključena: (i) svježa vina, (ii) desertna vina, (iii) vina dozrela u barrique bačvama, te (iv) vina proizvedena tradicionalnom tehnologijom uz produljenu maceraciju i dozrijevanje. Materijali i metode Uzorci vina Uzorci vina Malvazije istarske uključeni u ovo istraživanje sakupljeni su od proizvođača iz hrvatske i slovenske Istre. Odabran je ukupno 41 uzorak: 23 reprezentativna uzorka vina tzv. svježe Malvazije iz berbe 2013., 3 uzorka desertnog vina Malvazije, dobivenog posebnim postupcima prosušivanja grožđa (berbe i 2011.) od kojih su neki dozrijevali u drvenim bačvama, 11 uzoraka vina Malvazije dozrelog u barrique bačvama od hrastovog drveta (berbe ), te 4 uzorka vina tzv. tradicionalne Malvazije proizvedene uz produljenu fermentativnu i post-fermentativnu maceraciju masulja i dozrijevanje u bačvama od hrastovog drveta (berbe 2009., i 2013.). Analiza hlapljivih spojeva arome Za izdvajanje minornih hlapljivih spojeva iz uzoraka vina uporabljen je SPME uređaj za mikroekstrakciju na čvrstoj fazi proizvođača Supelco (SAD) s vlaknom presvučenim DVB/Carboxen/PDMS (50/30 µm) fazom (Supelco, SAD). Njihova identifikacija i kvantifikacija provedena je uporabom vezanog sustava plinski kromatograf Varian 3900 / spektrometar masa Varian Saturn 2100T (Varian, SAD), opremljenog kolonom Rtx-WAX (duljina 60 m, 0,25 mm ID, 0,25 µm d.f.; Restek, SAD). Etil-acetat, metanol i viši alkoholi analizirani su nakon izravnog ubrizgavanja destilata uzoraka vina u plinski kromatograf Varian 3350 (Varian, SAD), opremljen s plameno-ionizacijskim detektorom i RTX-Wax kolonom. Statistička obrada podataka Dobiveni podaci kromatografske analize obrađeni su kemometrijski, analizom glavnih sastavnica (eng. Principal Component Analysis; PCA) i linearnom diskriminantnom analizom (eng. Linear Discriminant Analysis; LDA) th Croatian and 10 th International Symposium on Agriculture

3 Diferencijacija različitih stilova vina Malvazija istarska na osnovi aromatskog profila Rezultati i rasprava U uzorcima različitih stilova vina Malvazije istarske izučavanih u ovom radu identificirano je i kvantificirano ukupno 50 hlapljivih spojeva arome. Primjena analize glavnih sastavnica (PCA) na skup sastavljen od 41 uzorka i 13 spojeva primarne arome nije u potpunosti razdvojila uzorke Malvazije prema stilu vina, ali su uočene jasne tendencije (Slika 1). Slika 1. Razdvajanje uzoraka vina Malvazije istarske prema stilu na osnovi profila hlapljivih spojeva primarne arome, monoterpena i C 13 -norizoprenoida, u smjerovima prve dvije glavne sastavnice PC1 i PC2 (a) hlapljivi spojevi primarne arome b) uzorci vina Uzorci svježih vina bili su okarakterizirani višim koncentracijama glavnih monoterpenola, kao i najvažnijeg norizoprenoida β-damascenona, te prilično razdvojeni od Malvazija proizvedenih dozrijevanjem u drvenim bačvama (barrique i tradicionalna tehnologija) uzduž PC1. Dozrele Malvazije grupirane su ka pozitivnim vrijednostima PC1 zahvaljujući višim udjelima α-terpineola i ostalih norizoprenoida, što je u skladu s podatkom da α-terpineol nastaje oksidacijom drugih monoterpena tijekom dozrijevanja (Ferreira i sur., 1997), te da postoji snažna korelacija između starosti vina i sadržaja vitispirana, aktinidola i 1,1,6-trimetil-1,2-dihidronaftalena (TDN) (Loscos i sur., 2010). Uzorci desertnih vina razdvojeni su od ostalih stilova na osnovi viših udjela monoterpena (koncentriranih tijekom prosušivanja) uzduž smjera PC2, ali i razasuti uzduž PC1 ovisno o starosti vina. U slučaju hlapljivih spojeva sekundarne arome (Slika 2), uzorci svježih Malvazija bili su povezani s višim koncentracijama većine fermentacijskih etilnih i acetatnih estera. Poznato je da navedeni spojevi, posebno esteri, hidroliziraju tijekom dozrijevanja, te su stoga niže koncentracije u uzorcima ostala tri stila vina očekivane (Câmara i sur., 2006; Garde Cerdán i sur., 2002). Uzorci vina Malvazije tradicionalne tehnologije, koja uključuje i produljenu maceraciju, grupirani su vrijednostima PC1 i PC2 koje odgovaraju višim koncentracijama metanola i viših alkohola. Section 8. Viticulture and Enology 517

4 Igor LUKIĆ, Sanja RADEKA, Andreja VANZO, Klemen LISJAK, Dejan BAVČAR Slika 2. Razdvajanje uzoraka vina Malvazije istarske prema stilu na osnovi profila hlapljivih spojeva sekundarne arome, hlapljivih kiselina, viših alkohola i hlapljivih estera, u smjeru prve dvije glavne sastavnice PC1 i PC2 (a) hlapljivi spojevi sekundarne arome b) uzorci vina Povišeni metanol očekivana je posljedica maceracije, dok je porast viših alkohola moguće rezultat dulje maceracije uz doticaj s kisikom i više temperature fermentacije (Vidrih i Hribar, 1999). Viša koncentracija etil-acetata u dozrelim vinima rezultat je djelovanja bakterija octene kiseline tijekom prosušivanja (desertna vina) i dozrijevanja u aerobnim uvjetima (Câmara i sur., 2006). Spomenuta vina grupirana su ka pozitivnim PC1 vrijednostima i na osnovi povišenih udjela etil-2- i etil-3-metil-butanoata, te etil-laktata i dietil-sukcinata, estera koji nastaju malolaktičnom fermentacijom i esterifikacijom tijekom dozrijevanja (Câmara i sur., 2006; Garde Cerdán i sur., 2002). Analizom glavnih sastavnica (PCA) obrađen je i skup od 41 uzorka i 12 hlapljivih spojeva tercijarne arome (Slika 3). Jedini izučavani spoj karakterističan za svježa vina bio je 4-vinil-guaiakol, koji nastaje uglavnom u standardnoj alkoholnoj i eventualno malolaktičnoj fermentaciji (Garde Cerdán i sur., 2002; Suárez i sur., 2007). Slika 3. Razdvajanje uzoraka vina Malvazije istarske prema stilu na osnovi profila hlapljivih spojeva tercijarne arome, hlapljivih fenola, laktona i furana, u smjeru prve dvije glavne sastavnice PC1 i PC2 (a) hlapljivi spojevi tercijarne arome b) uzorci vina th Croatian and 10 th International Symposium on Agriculture

5 Diferencijacija različitih stilova vina Malvazija istarska na osnovi aromatskog profila S druge strane, povišeni 4-etil-fenol i 4-etil-guaiakol u dozrelim vinima uglavnom su posljedica djelovanja sojeva Brettanomyces i Dekkera tijekom dozrijevanja (Perestrelo i sur., 2011), gdje je redukcija vinil- u etilfenole katalizirana aerobnim uvjetima i višom temperaturom (Garde Cerdán i sur., 2002). Iako su pojedini γ-laktoni identificirani u svježim vinima, njihove koncentracije uglavnom rastu zahvaljujući dozrijevanju u bačvama zbog ekstrakcije iz drveta. Jedini izvor tzv. hrastovih laktona u vinu je drvo hrasta (Perestrelo i sur., 2011), te je logično da su navedeni spojevi karakteristični za vina dozrela u drvu. Slično vrijedi i za furane koji također u većim količinama nastaju tijekom dozrijevanja (Perestrelo i sur., 2011). U svrhu pronalaženja potencijalnih biljega različitih stilova vina Malvazije istarske, ukupni skup podataka obrađen je linearnom diskriminantnom analizom (LDA). Sukladno Wilksovom lambda kriteriju, model je odabrao 12 hlapljivih spojeva uz uspješno diferenciranje uzoraka prema stilu. Ukupno je postignuta točna klasifikacija od 97,56 %: uzorci svježe, desertne i tradicionalne Malvazije diferencirani su uz 100 %-tnu točnost, dok je postotak za barrique stil bio 90,9 %. Odabrani spojevi bili su, redom: dietil-sukcinat, 4-etil-fenol, citronelol, furfural, cis-hrastov lakton, γ-undekalakton, aktinidol eter (izomer I), trans-hrastov lakton, etilizobutanoat, etil-3-metil-butanoat, β-ionon i γ-dekalakton. Potrebno je napomenuti da su u ovom slučaju u model uključeni oni spojevi koji najbolje međusobno diferenciraju sva četiri stila. Kao posljedica toga dobiveni su rezultati koji se mogu učiniti nesvrsishodnim, te zbog toga nisu prikazani grafički (npr. diskriminantna analiza u model nije uključila spojeve sekundarne arome koji bi izvrsno izdvojili svježa vina, ali su relativno neučinkoviti za međusobnu diferencijaciju ostala tri stila). Zaključci U ovom istraživanju pokazano je da se četiri glavna stila Malvazije istarske u svojoj osnovi razlikuju prema aromatskom profilu, što se u konačnici zasigurno snažno odražava na razlike u njihovim senzornim svojstvima. Svježa vina Malvazije istarske uglavnom su karakterizirana tipičnim aromatskim profilom bijelih vina uz dominaciju hlapljivih estera i kiselina, kojih sadržavaju osjetno više nego vina ostalih izučavanih stilova. U odnosu na Malvazije dozrele u barrique bačvama i tradicionalnom tehnologijom, sadrže i više koncentracije većine glavnih monoterpena i β-damascenona. Desertna vina obilježena su povišenim udjelima glavnih monoterpenola i etil-acetata kao posljedice prosušivanja grožđa, ali u pojedinim slučajevima, ovisno o starosti, i tercijarnim aromama. Vina iz barrique bačava te ona tradicionalna, u većini slučajeva nisu jasno međusobno razlučiva jer sadrže povišene koncentracije određenih norizoprenoida, etil-acetata, etil estera kiselina kratkog razgranatog lanca, estera diprotonskih i hidroksi-kiselina, hlapljivih fenola, laktona i furana, čime su pak vrlo dobro diferencirana od svježih vina. Tradicionalna vina Malvazije istarske odlikovala su se povišenim koncentracijama metanola i viših alkohola. Iako vrlo dobra, nepotpuna diferencijacija različitih stilova vina Malvazije istarske u pojedinim slučajevima uglavnom je posljedica određene heterogenosti proizašle iz različitih kombinacija ključnih faktora utjecaja (berba i starost, tehnološke varijacije, i sl.), te djelomičnog preklapanja pojedinih karakteristika. Dobiveni rezultati predstavljaju dobru osnovu za razumijevanje specifičnih senzornih svojstava četiri glavna stila vina Malvazije istarske, bolje upravljanje njihovom kvalitetom, te daljnju diferencijaciju prema potrebama i zahtjevima potrošača. Istraživanja u ovom radu dio su projekta Putevima Malvazije istarske (Malvasia TourIstra) u sklopu Operativnog programa prekogranične suradnje IPA Slovenija - Hrvatska , financiranog od strane Europskog fonda za regionalni razvoj. Autori zahvaljuju proizvođačima vina Malvazije istarske koji su donirali uzorke. Literatura Bavčar, D., Baša Česnik, H., Čuš, F., & Košmerl, T. (2011). The influence of skin contact during alcoholic fermentation on the aroma composition of Ribolla Gialla and Malvasia Istriana Vitis vinifera (L.) grape wines. International Journal of Food Science & Technology, 46, Câmara, J.S., Alves, M.A., & Marques, J.C. (2006). Changes in volatile composition of Madeira wines during their oxidative ageing. Analytica Chimica Acta, 563(1 2), Ferreira, V., Escudero, A., Fernández, P., & Cacho, J.F. (1997). Changes in the profile of volatile compounds in wines stored under oxygen and their relationship with the browning process. Zeitschrift für Lebensmittel Untersuchung und Forschung A, 205, Section 8. Viticulture and Enology 519

6 Igor LUKIĆ, Sanja RADEKA, Andreja VANZO, Klemen LISJAK, Dejan BAVČAR Garde Cerdán, T., Rodríguez Mozaz, S., & Ancín Azpilicueta, C. (2002). Volatile composition of aged wine in used barrels of French oak and of American oak. Food Research International, 35, Loscos, N., Hernández-Orte, P., Cacho, J., & Ferreira, V. (2010). Evolution of the aroma composition of wines supplemented with grape flavor precursors from different varietals during accelerated wine ageing. Food Chemistry, 120, Lukić, I., Plavša, T., Sladonja, B., Radeka, S., & Peršurić, Đ. (2008). Aroma compounds as markers of wine quality in the case of Malvazija istarska young wine. Journal of Food Quality, 31, Perestrelo, R., Barros, A.S., Câmara, J.S., & Rocha, S.M. (2011). In-Depth Search Focused on Furans, Lactones, Volatile Phenols, and Acetals As Potential Age Markers of Madeira Wines by Comprehensive Two-Dimensional Gas Chromatography with Time-of-Flight Mass Spectrometry Combined with Solid Phase Microextraction. Journal of Agricultural and Food Chemistry, 59, Radeka, S., Herjavec, S., Peršurić, Đ., Lukić, I., & Sladonja, B. (2008). Effect of Different Maceration Treatments on Free and Bound Varietal Aroma Compounds in Wine of Vitis vinifera L. cv. Malvazija istarska bijela. Food Technology and Biotechnology, 46, Suárez, R., Suárez-Lepe, J.A., Morata, A., & Calderón, F. (2007). The production of ethylphenols in wine by yeasts of the genera Brettanomyces and Dekkera: A review. Food Chemistry, 102, Vidrih, R., & Hribar, J. (1999). Synthesis of higher alcohols during cider processing. Food Chemistry, 67(3), sa2015_p th Croatian and 10 th International Symposium on Agriculture

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