The effects of Saccharomyces cerevisiae strains carrying alcoholic fermentation on the fermentative and varietal aroma profiles of young and aged Tempranillo wines

Food Chemis y: X 9 (2021) 100116 A ailable online 9 Feb ua y 2021 2590-1575/© 2021 The Au ho (s). Published by Else ie L d. This is an open access a icle unde he CC BY license (h p://c ea i ecommons.o g/licenses/by/4.0/). The e ec s o Saccha omyces ce e isiae s ains ca ying alcoholic e men a ion on he e men a i e and a ie al a oma p o iles o young and aged Temp anillo wines Ma ie Dena a , Dolo es P´ e ez b , c , d , Jos´ e Ma ía He as b , Ampa o Que ol c , Vicen e Fe ei a a , * a Labo a o y o A oma Analysis and Enology (LAAE), Depa men o Analy ical Chemis y, Uni e sidad de Za agoza, Ins i u o Ag oalimen a io de A ag´ on (IA2) (UNIZAR-CITA), c/Ped o Ce buna 12, 50009 Za agoza, Spain b Lallemand Bio S.L., 08028 Ba celona, Spain c Ins i u e o Ag ochemis y and Food Technology (IATA-CSIC), 46980 Pa e na, Valencia, Spain d Es aci´ on Expe imen al Ag opecua ia Mendoza (EEA), Ins i u o Nacional de Tecnología Ag opecua ia (INTA), 5507 Luj´ an de Cuyo, Mendoza, A gen ina ARTICLE INFO Keywo ds: Fe men a ion Longe i y F ui y es e s Vola ile phenols No isop enoids Vanillin S ecke aldehydes E hyl leuca e ABSTRACT Ten di e en Saccha omyces ce e isiae s ains e men ed semi-syn he ic mus s con aining a Polyphenolic and A oma P ecu so F ac ion (PAF) ex ac ed om Temp anillo g apes. A oma compounds we e s udied by Gas Ch oma og aphy (GC), GC-Ol ac ome y and GC-Mass Spec ome y (MS), du ing e men a ion by apping ola ilized a oma, immedia ely a e e men a ion and a e accele a ed aging. Vola iles los by e apo a ion du ing e men a ion a e mos ly e men a i e compounds and no g ape- ela ed odo an s. Isobu anal and some es e s a e mos ly los du ing e men a ion. In many cases he impac o yeas s ain is e iden only a e aging. S ains could be classi ied in o 3 majo clus e s wi h ma ked di e ences in e men a i e and a ie al p o iles. Linalool and ge aniol we e ound o ha e e men a i e o igin. S. ce e isiae yeas s ains can e ec i ely modula e a ie al a oma, likely h ough speci ic enzyma ic ac i i ies ac ing on g ape phenolic acids and no isop enoid a oma p ecu so s and may be speci ically used o mi iga e some aging- ela ed o odou s, such as massoia lac one, guaiacol o TDN. 1. In oduc ion Wine a oma is i s mos ou s anding senso y p ope y and is essen ial o i s quali y and di e en ia ion (Cha e s & Pe ig ew, 2007). Al hough he numbe o ola ile molecules which can be a pa o he ola ile ac ion o wines is e y la ge, exceeding mos likely se e al housands, i has been sugges ed ha 70 di e en odou chemicals a e hose playing majo oles on he a oma ic p ope ies o wines (De-la- Fuen e-Blanco, S´ aenz-Na ajas, & Fe ei a, 2020). Quan i a i ely, he mos abundan wine odo an s a e alcoholic e men a ion (AF) by-p oduc s, pa icula ly highe alcohols, e hyl es e s and ace a es, some ca bonyls and acids. By con as , many o he ele- an odo an s de i e om g ape speci ic p ecu so s and can be p esen a e y limi ed concen a ions, wi hin he ng/L ange in he case o poly unc ional me cap ans, ew hund eds o ng/L in he case o β-ion- one, a ound he ew μ g/L in he case o β-damascenone o below 0.2 mg/ L in he cases o e penols, ola ile phenols and anillin de i a i es (Fe ei a & Lopez, 2019; Ruiz e al., 2019). The e a e also ele an di e ences be ween compounds ega ding he ime a which hey a e o med o libe a ed. Mos e men a ion by- p oduc s de i e om yeas amino acid and a y acid me abolisms (Swiege s, Ba owsky, Henschke, & P e o ius, 2005), so ha hey a e signi ican ly o med om he ea ly s ages o AF and a e known o be pa ly los by e apo a ion (G´ omez-Plaza, Ma ínez-Cu illas, & Laencina, 1993; Gue ini e al., 2016). Some o he compounds, such as he mon- o e penoids ge aniol and linalool, a e di ec ly libe a ed om g ape-speci ic glycosyla ed p ecu so s du ing e men a ion by he ac ion o yeas β-glucosidases (Guna a, Bi eu , B illoue , Bayono e, & Co - donnie , 1988). Poly unc ional me cap ans a e also byp oduc s o AF and may be also pa ially los by e apo a ion du ing e men a ion. On he con a y, he o ma ion and accumula ion o some o he ola iles equi es aging ime. In ac , a quan i a i ely ele an g oup o a oma compounds and a oma p ecu so s a e subjec ed o se e al slow chemical eac ions such as acid hyd olysis, es e i ica ion o in a- * Co esponding au ho . E-mail add ess: [email p o ec ed] (V. Fe ei a). Con en s lis s a ailable a ScienceDi ec Food Chemis y: X jou nal homepage: www.sciencedi ec .com/jou nal/ ood-chemis y-x h ps://doi.o g/10.1016/j. ochx.2021.100116 Recei ed 20 Ma ch 2020; Recei ed in e ised o m 24 Decembe 2020; Accep ed 19 Janua y 2021 Food Chemis y: X 9 (2021) 100116 2 molecula ea angemen s ha will g ea ly a ec wine a oma p o ile. This is he case o mos no isop enoids and anillin de i a i es, such as β-damascenone and TDN o anillin and ace o anillone, whose le els inc ease du ing aging as he consequence o di e en ans o ma ions o g ape ca o enoid me aboli es (Win e hal e & G¨ ok, 2013) o o g ape glycosyla ed p ecu so s (Fe ei a & Lopez, 2019). No ably, i has been ecen ly obse ed ha he yeas gene a plays a majo ole in he mod- ula ion o TDN concen a ion in wine a e some ime o aging (Oli ei a & Fe ei a, 2019). A hi d g oup o odo an s su e ing changes du ing aging a e labile molecules, such as linalool o ge aniol, which deg ade du ing aging o o m α - e pineol, ne ol o 1,8-cineole (Wa e house, Sacks, & Je e y, 2016). O he g oups o odo an s deeply a ec ed by aging a e ui y es e s and ace a es. The ace a es o highe alcohols a e quickly hyd olysed and hei le els soon ade away. On he con a y, he e hyl es e s o b anched acids - isobu y ic, 2-me hylbu y ic and iso- ale ic acids - and o o he mino acids, slowly and s eadily inc ease by es e i ica ion o hei p ecu so acids wi h e hanol (Díaz-Ma o o, Schneide , & Baumes, 2005). The exis ence o all hese p ocesses makes ha aging ime should be conside ed as an impo an ac o o assess he ole o yeas s on wine a oma modula ion. Saccha omyces ce e isiae is he mic o-o ganism mos widely s udied ega ding i s senso y impac on wine (Temp` e e e al., 2018). Howe e , mos p e ious s udies ha e deal wi h he sho - e m impac o his yeas on bo h e men a i e and a ie al a oma p o iles (Game o, He n´ andez- O e, Que ol, & Fe ei a, 2011; Gammacu a, Ma chand, Moine, & de Re el, 2017; Molina e al., 2009), neglec ing aging e ec s and also possible di e ences in he a oma p o iles los by e apo a ion, which could be pa icula ly ele an in lab-scale e men a ions. Bo h aspec s will be specially add essed in he p esen wo k whose objec i e is o e alua e he impac o S. ce e isiae yeas s ains on he e men a i e and a ie al a oma o Temp anillo wine du ing and a e e men a ion, and a e a pe iod o accele a ed aging. 2. Ma e ial and me hods 2.1. Reagen s and s anda ds Dichlo ome hane and me hanol (≥99%) Dis o-Pes icide esidue g ade we e supplied by Me ck (Da ms ad , Ge many). ACS quali y ab- solu e e hanol was pu chased om Pan eac (Ba celona, Spain). 2- Bu anol (≥99%), 4-me hyl-2-pen anol (99%), 4-hyd oxy-4-me hyl-2- pen anone (99%), e hyl hep anoa e (99%) and hep anoic acid (99%) we e used as in e nal s anda ds o majo compounds analysis. 2-Oc a- nol (99.5%), 3-oc anone (99%) and 3,4-dime hylphenol (99%) we e used as in e nal s anda ds o mino compounds analysis. They we e pu chased om Me ck as well as he chemical s anda ds used in his s udy (>98%), excep o TDN which was syn hesized by Synchem UG &Co (Felsbe g, Ge many) wi h a pu i y o 80%. LiCh olu EN esin ca idges we e ob ained om Me ck (Da ms ad , Ge many). Sep Pak- C18 esins, p epacked in 10 g ca idges we e om Wa e s (I eland). 2.2. Semi-syn he ic mus p epa a ion 2.2.1. P ecu so s and phenols ex ac ion om Temp anillo g apes Phenolic and A oma p ecu so F ac ion (PAF) was ex ac ed om Temp anillo g apes mis elle p epa ed a he ICVV o Log o˜ no (Spain) ollowing he p ocedu e de eloped by Aleg e e al. (2020). 2.2.2. Syn he ic g ape mus Syn he ic mus was p epa ed as desc ibed in He n´ andez-O e, Bely, Cacho, and Fe ei a (2006). Ni ogen con en was adjus ed by mixing 220 mg/L o (NH 4 ) 2 HPO 4 , and a mix u e o amino acids esembling he a e age p o ile o Temp anillo g ape a ie y (He n´ andez-O e, Cacho, & Fe ei a, 2002). Syn he ic glu a hionyla ed and cys einila ed (Cys) p ecu so s o 3-me cap ohexanol (MH) and 4-me cap open an-2-one (MMP) we e added om a solu ion in MilliQ wa e , (0.1 mg/L Cys- MH, 0.05 mg/L Cys-MMP, 1 mg/L Glu-MH, 0.05 mg/L Glu-MMP). A e pH adjus men o 3.5 wi h NaOH, syn he ic mus was s e ilized by il a ion (0.45 μ m) inside a e ical lamina low chambe . PAF was dealcoholized, esuspended in s e ile dis illed wa e and added o syn- he ic mus a 10% ( / ). 2.3. Wine elabo a ion 2.3.1. Yeas s s ains, p e-cul u e condi ions and yeas g ow h moni o ing Ten S. ce e isiae s ains (Lallemand Bio SL, Mad id, Spain) we e used: Lal in ICV D254™ (D254), Lal in Clos™ (CLOS), U a e m HPS™ (HPS), Eno e m BDX™ (BDX), Lal in Rhˆ one2056® (RHONE), Lal in ICV D80™ (D80), Lal in 71B™ (71B), Lal in Pe sy™ (PERSY), Lal in ICV OKAY™ (OKAY), IONYS w ™ (IONYS). Ac i e d y yeas s we e ehyd a ed in dis illed wa e a 37 ◦C o 20 min unde agi a ion, 100 μ L o hese cul u es we e pla ed on Glucose Pep one Yeas (GPY) and incuba ed a 25 ◦C o 24 h. P e-cul u es we e p epa ed by inocula ing se e al col- onies in o 5 mL o GPY b o h a 25 ◦C o 24 h. A e quick cen i uga- ion, GPY b o h was disca ded and colonies we e esuspended in dis illed wa e . Li ing cells we e quan i ied by low cy ome y and each e men e o syn he ic mus was inocula ed a 10 6 li ing cells/mL. Cell iabili y and i ali y we e moni o ed by low cy ome y (Tilloy, O iz- Julien, & Dequin, 2014). 2.3.2. Expe imen al design All he e men a ions we e ealized in iplica es. Fe men a ion wi h CLOS and IONYS we e epea ed in mus wi hou PAF addi ion, whose olume was eplaced by s e ile dis illed wa e . Un e men ed con ol o syn he ic mus wi h and wi hou PAF was also included in duplica es. Expe imen al p ocedu e is summa ized in Fig. S1. 2.3.3. Fe men a ion sys em Fe men a ions we e ca ied ou in 100 mL glass e men e s con- aining 50 mL o syn he ic mus so ha headspace ep esen ed 50% o he o al olume. Fe men e s we e igh ly closed wi h a pe o a ed sili- cone co k in which an ai lock (Mic omal a S.L., Mad id, Spain) was inse ed. Needles and 5 mL sy inges we e inse ed in o he silicon co k o allow sampling wi hou opening he e men e . 2.3.4. Fe men a ion moni o ing Fe men a ions we e ca ied ou a 25 ◦C, unde agi a ion a 150 pm using a magne ic s i e . CO 2 elease was moni o ed by weighing. The main chemical e men a i e pa ame e s we e analysed a he beginning and a he end o e men a ion, including suga s, acids and alcohols by UHPLC as desc ibed in Su e al. (2019). 2.3.5. End o e men a ion and accele a ed aging A he end o e men a ion, samples we e cen i uged and condi- ioned o accele a ed anoxic aging in o a ee-O 2 chambe Jacomex (Dagneux, F ance). Samples we e placed in o 18 mL glass ubes capped wi h non-me allic sc ew caps and bagged in high densi y plas ic bags con aining oxygen sca enge s Anae oGenTM (The mo Scien i ic, USA) and incuba ed a 50 ◦C o 5 weeks. This me hodology was de eloped by Vela, He n´ andez-O e, F anco-Luesma, & Fe ei a, (2017) and was success ully applied o he aging o e men ed media (Oli ei a & Fe - ei a, 2019). I was obse ed ha 5 weeks o anoxic aging a 50 ◦C we e oughly equi alen o one yea o aging a oom empe a u e. 2.4. Analysis o he a oma e apo a ed du ing e men a ion A p e-pu i ied s anda d SPE ca idge illed wi h 160 mg LiCh olu - EN esin was lodged in o he ai lock o he e men e s o ap ola iles emi ed du ing e men a ion. When his inished, he ca idge was emo ed, d ied and elu ed wi h 1.6 mL o dichlo ome hane con aining 5% ( / ) o me hanol. Fo GC-Ol ac ome y (GC-O), a single ex ac was p epa ed by M. Dena e al. Food Chemis y: X 9 (2021) 100116 3 mixing 100-uL o each one o he o y ex ac s ob ained om he 40 di e en e men a ions. The ex ac was ca e ully concen a ed by e apo a ion o he sol en unde ni ogen low un il a 0.2 mL inal olume and injec ed in o he GC-O sys em, as desc ibed in San-Juan, Pe ’ka, Cacho, Fe ei a, & Escude o, (2010). Ol ac ome ic sco es we e ob ained by combining in ensi y and equency as desc ibed by D a - nieks, (1985). The odo an s we e iden i ied by compa ison o hei de- sc ip o s, ch oma og aphic e en ion index in DB-WAX and DB-5 columns and mass spec a wi h hose o pu e e e ence compounds. Fo quan i ica ion o he odo an s p esen in he ex ac s, hese we e spiked wi h he in e nal s anda ds, concen a ed by e apo a ion unde ni ogen up o 0.2 mL and analyzed. Majo e men a i e compounds we e di ec ly quan i ied by GC-FID analysis unde he condi ions desc ibed in O ega, L´ opez, Cacho, & Fe ei a, (2001). Mino com- pounds we e quan i ied by GC–MS analysis o he ex ac s in a Shimadzu QP2010 (Quio o, Japan) equipped wi h a DB-WAXe GC-column om Agilen (San a Cla a CA, USA), 30 m ×0.25 nm, 0.5 μ m o ilm hickness, p eceded by a 2 m ×0.25 mm uncoa ed p e-column. Ca ie gas was Helium a 1.26 mL/min. Injec ion olume was 1 μ L in spli mode ( a io 1/30). Injec o empe a u e was 250 ◦C. Ch oma og aphic o en em- pe a u e was ini ially a 30 ◦C o 1 min, hen aised a 1 ◦C/min o 35 ◦C, held o 1 min, hen a 1 ◦C/min o 40 ◦C, a 15 ◦C/min o 55 ◦C, held o 5 min, a 15 ◦C/min o 72 ◦C, held o 5 min, a 15 ◦C/min o 150 ◦C, held o 15 min. The Ion sou ce was kep a 220 ◦C and he in e ace a 230 ◦C. The mass analyse was se in single ion moni o ing mode. The comple e lis compounds quan i ied in bo h p ocedu es, including he m/z a ios selec ed o MS quan i a ion is a ailable in Table S1. 2.5. Analysis o majo and mino ola ile compounds Majo me aboli es o alcoholic e men a ion (highe alcohols and hei ace a es, ola ile a y acids and hei e hyl es e s, b anched a y acids and hei e hyl es e s, ace oin, diace yl, and ace aldehyde), usually p esen in wines a le els abo e 0.2 mg/L, we e analysed by he GC-FID analysis o a dichlo ome hane mic oex ac as desc ibed by O ega, L´ opez, Cacho, & Fe ei a, (2001). Mino a oma compounds p esen in wine a le els 0.1–200 μ g/L (b anched e hyl es e s, e penes, no isop enoids, anillin de i a i es, ola ile phenols) we e analysed by GC–MS analysis o a SPE ex ac ollowing he p o ocol desc ibed by L´ opez, Azna , Cacho, & Fe ei a, (2002). The comple e lis o quan i ied is a ailable in Table S1. 2.6. S a is ical analysis S a is ical analysis and g aphics we e ealized using R so wa e (RS udio Team, 2020). Signi icance was de e mined by analysis o a iance (ANOVA) using ano a unc ion om ca package ( .3.0.2) and Tuckey’s hones ly signi icance di e ence es was pe o med on ANOVA esul s, using HSD. es unc ion om ag icolae package ( .1.3.1). P in- cipal Componen Analysis (PCA) we e pe o med and plo ed using ac oex a package ( .1.0.5). 3. Resul s 3.1. Odo an s los du ing e men a ion In o de o assess he ype and amoun s o odo an s los du ing e men a ion, a small ap was ins alled in he e men e s. A GC-O sc eening p ocedu e was ca ied ou on an ex ac ob ained by mixing small aliquo s o all he ex ac s ob ained in he expe imen . Resul s a e summa ized in Table 1. O e all, wen y- wo odo an s we e de ec ed wi h GC-O sco es abo e 20%. Ele en ou o he wen y- wo odo an s we e iden i ied a maxima le el o con idence. In i e o he cases, some o he iden i y c i e ia could no be comple ely ul illed because o di e en easons, such as excessi ely low le els o ge a good MS spec um (Z and E-2-nonenals), co-elu ion in he non-pola column (c esol and guaiacol) o disc epancy in he odou (e hyl oc ano e). The iden i ica ion o he six less in ense odo an s was based only on he coincidence o he odou and RI o he odo an in he pola column wi h hose o he s anda d, and should be conside ed en a i e. The mos in ense odo an s we e mainly by-p oduc s o alcoholic e men a ion: isoamyl alcohol and 2-phenyle hanol, 3-me hylbu anal, and a nume ous g oup o es e s (isop opyl and isoamyl ace a es, and he e hyl es e s o isobu y ic, bu y ic, hexanoic, oc anoic and decanoic acids). Apa om hese, c esol and Z and E-2-nonenals we e also be- ween he wel e mos in ense. The o igin o c esol is no clea , bu i could be a b eakdown p oduc o he polyphenols p esen in e men a- ion media; while nonenals a e known de i a i es o he au o-oxida ion o g ape a y acids. Resul s, he e o e, con i m ha he mos ele an odo an s pu ged ou du ing e men a ion a e by-p oduc s de i ed om yeas me abolism o g ape a y acid au o-oxida ion and no a ie al a oma compounds eleased om speci ic a oma p ecu so s in g ape, such as e penols, no isop enoids o poly unc ional me cap ans. 3.2. Quan i a i e assessmen o he ola iles los du ing e men a ion The amoun s o eigh een odo an s apped in he ca idges ins alled in he PAF-con aining e men e s a e summa ized in Table 2. In gene al, le els we e low and in some cases we e a ec ed by high imp ecision. The o al mass o ola iles apped in he ca idges anged om a ound 1 mg (D80) o a ound 2 mg (71B). The majo ola ile was iso- bu y aldehyde, which accoun s o mo e han 50% o he o al ola iles apped. Fou o he odo an s, isop opyl ace a e, isoamyl ace a e, iso- bu anol, and isoamyl alcohol can be also ound a le els abo e 100 μ g. The le els o some ola iles eleased we e signi ican ly ela ed o he Table 1 Iden i ica ion o odo an s pu ged ou du ing e men a ions and apped in LiCh olu -EN ca idges placed be o e he Mulle al es o he e men e s. The GC-O expe imen was ca ied ou on an ex ac made by mixing he elua es o he di e en aps. Re en ion indexes (RI) in DB-WAX and DB-5 columns, iden i i- ca ions, ol ac ome ic sco es (MF%) and odo desc ip o s a e p esen ed in he able. Compounds a e ma ked by le e s acco ding o he eliabili y o hei iden i ica ion (see legend). RI (DB- WAX) RI (DB- 5) Compound* MF (%) Odo desc ip ion 1220 <900 isoamyl alcohol A 87 cheese, ancid 940 <900 3-me hylbu anal A 78 cheese, ancid 935 <900 isop opyl ace a e A 62 ui y, s awbe y 975 <900 e hyl isobu y a e A 62 ui y, s awbe y 2097 c esol B 55 phenolic, lea he 1929 1115 2-phenyle hanol A 53 lo al, ose 1039 <900 e hyl bu y a e A 46 ui y, s awbe y 1241 988 e hyl hexanoa e A 45 ui y 1513 1133 Z-2-nonenal B 38 ancid, cucumbe 1128 <900 isoamyl ace a e A 37 banana 1441 1193 e hyl oc anoa e B 35 mush oom, plas ic, humidi y 1544 1153 E-2-nonenal B 29 a , ancid 1621 1391 e hyl decanoa e A 29 soap 1696 <900 2 and 3-me hylbu y ic acid A 26 swea , ancid 1842 guaiacol B 26 smoky, bu n 966 <900 e hyl p opanoa e A 24 ui y 1425 1-nonen-3-one C 24 mush oom, unde g ow h 1468 decanal C 24 g ass, lo al 1953 Z-whisky lac one C 24 spicy 2169 2-phenoxye hanol C 24 ancid, ca on 2245 4- inylguaiacol C 24 spicy 2214 so olon C 22 spicy * A: iden i ica ion conclusi e. Expe imen al RIs in wo columns, odou and MS co esponded o he one ob ained wi h he pu e chemical s anda d; B: iden i y highly likely, one o he p e ious c i e ia ( wo RIs, odou , MS) ailed; C: en a i e iden i ica ion based on RI on a single column, odou and p e ious li e a u e. M. Dena e al. Food Chemis y: X 9 (2021) 100116 4 yeas s ain, al hough in mos cases di e ences we e no e y high. Majo di e ences co espond o he s ain IONYS, whose ca idges con ained highes le els o ace a es and o some e hyl es e s, ollowed by he s ain 71B, wi h maxima le els o aldehydes. As expec ed, ola iles pu ged du ing e men a ion a e mos ly non- pola a oma compounds such as es e s, ace a es o ca bonyls. Only wo acids and wo alcohols we e ound amongs he quan i iable ola ile compounds. In hese cases, he amoun s e apo a ed co esponded o e y small ac ions o he ola iles p oduced. In he pa icula case o isoamyl alcohol, he 664 μ g ound in he ca idge o OKAY s ain co espond o 13 mg/L in he 50 mL o liquid. Conside ing ha he ecen ly e men ed wine con ained a ound 190 mg/L o his compound (Table 2, Supplemen a y ma e ial), i can be es ima ed ha less han 5% o he o al amoun o isoamyl alcohol p oduced was e apo a ed. On he opposi e side, isobu y aldehyde was ound in he ca idge om he s ain 71B a 1.29 mg, which amoun s o 26 mg/L in he 50-mL olume, while epo ed le els o his compound in wine a e well below 0.1 mg/L (Culle ´ e, Cacho, & Fe ei a, 2007). This sugges s ha isobu y aldehyde is a majo e men a ion ola ile which is nea ly comple ely (>99%) los by e apo a ion. Simila conside a ions applied o isop opyl and isoamyl ace a es and e hyl p opanoa e indica e ha mo e han 70% o hese a omas a e los by e apo a ion. Le els pu ged ou o highe es e s, such as e hyl bu y a e, isobu y a e, hexanoa e, oc anoa e and decanoa e, and o highe aldehydes, such as 2 and 3-me hylbu anal, we e mo e modes . Ye , hey ep esen signi ican ac ions o he o al o med. In he cases o e hyl hexanoa e and decanoa e, o ins ance, he ac ions los a e 68 and 35% o he o al amoun s o med, espec i ely. In any case, esul s e eal ha he CO 2 eleased du ing e men a ion ca ied ou la ge amoun s o isobu y aldehyde, isop opyl ace a e, e hyl p opanoa e and isoamyl ace a e, which in ac a e mos ly los in his pe iod; and also signi ican amoun s in absolu e bu no in ela i e e ms, o isobu anol and isoamyl alcohol. Some o he es e s, such as e hyl bu y a e, hexanoa e, oc anoa e and decanoa e we e p oduced a much smalle le els bu ye , we e signi ican ly los by e apo a ion. 3.3. Majo e men a i e a oma compounds Twen y-six majo e men a ion ola iles we e de ec ed a concen- a ions supe io o de ec ion limi s in young wines (Table S2). In his case, i een ou o he wen y-six quan i ied ola iles we e signi ican ly ela ed o he yeas s ain. The mos di e en p o iles o ola iles we e ob ained in wines e men ed wi h IONYS, which p oduced maxima le els o mos e hyl es e s and ace a es, and also o iso ale ic acid, ace oin and γ-bu y olac one, and minima le els o ace ic and decanoic acids. O he s ains showing speci ic p o iles o ola iles we e 71B, BDX, D80, and PERSY. 71B p oduced maxima le els o me hionol, bu anol, Table 2 Amoun s (in μ g) o ola ile compounds pu ged ou du ing e men a ion and apped in LiCh olu -EN ca idges placed be o e he Mulle al es o he e men e s con aining 50 mL o syn he ic mus . The i s column gi es he signi icance o he ac o yeas on he le els o ola iles ound in he e men a ions ealized wi h PAF addi ion (p alues in bold a e in e io o 0.05). p alue MUST CLOS IONYS 71B BDX D254 D80 HPS OKAY PERSY RHONE Ace a es p opyl ace a e 6.24E- 10 0.10 ± 0.02 0.6 ±0.2 4.1 ±0.3 0.9 ±0.2 0.49 ± 0.06 0.6 ±0.2 0.5 ±0.2 0.9 ± 0.2 2.1 ±0.4 3.7 ±0.8 0.9 ±0.2 isop opyl ace a e 5.80E- 05 1 ±7 135 ±11 320 ±43 158 ±34 135 ±33 130 ±17 129 ±54 162 ± 43 199 ±18 250 ±33 171 ±23 isobu yl ace a e 4.47E- 02 n.d. 0.7 ±0.1 6 ±2 2.0 ±0.7 1.8 ±0.5 1.2 ±0.4 1 ±1 2.2 ± 0.6 1.4 ±0.4 3.1 ±0.1 1.1 ±0.5 isoamyl ace a e 4.70E- 04 15 ±4 6 ±3 104 ±18 23 ±17 – 17 ±11 8 ±15 18.2 ± 8.1 29 ±5 – 14 ±31 Acids 3-me hylbu y ic acid 1.14E- 01 n.d. 3 ±1 4 ±3 4 ±2 2.4 ±0.2 4 ±1 1.8 ±0.4 5.1 ± 2.0 2.7 ±3.0 3 ±2 4 ±1 2-me hylbu y ic acid 7.92E- 02 0.1 ±0.1 0.4 ±0.2 0.8 ±0.7 0.9 ±0.6 0.59 ± 0.09 0.9 ±0.4 0.41 ± 0.09 1.1 ± 0.5 1.4 ±0.8 0.6 ±0.6 1. 0 ±0.3 Alcohols isobu anol 4.69E- 02 74 ±20 130 ±24 129 ±8 128 ±4 – 176 ±30 142 ± 112 184 ± 31 127 ±54 – 120 ±16 isoamyl alcohol 1.57E- 01 387 ±13 420 ± 107 488 ±58 484 ± 226 – 497 ± 163 325 ± 318 507 ± 125 664 ± 116 – 364 ± 110 Ca bonyls ace aldehyde 3.07E- 03 5 ±2 3 ±2 3.6 ±0.6 14 ±10 – 3.3 ±0.8 4 ±1 4 ±1 4 ±2 – 5 ±2 isobu y aldehyde 6.39E- 02 n.d. 904 ± 482 684 ± 124 1291 ± 528 957 ± 464 548 ± 268 392 ± 210 692 ± 367 493 ± 267 644 ± 243 534 ± 113 2-me hylbu anal 5.85E- 02 n.d. 1.4 ±0.2 2.2 ±0.7 3 ±1 1.1 ±0.5 1 ±1 0.7 ±0.1 1.2 ± 0.8 2.4 ±0.8 1.7 ±0.1 0.9 ±0.9 3-me hylbu anal 2.05E- 01 n.d. 3.5 ±0.9 7 ±1 12 ±4 4.2 ±2.3 3 ±4 3. 0 ± 0.3 4 ±3 10 ±4 6.5 ±0.3 4 ±4 Es e s e hyl p opanoa e 5.28E- 04 0.1 ±0.2 6.5 ±0.6 38 ±18 5 ±1 4.9 ±0.3 6 ±7 5 ±2 7 ±2 7.5 ±0.5 10 ±3 6 ±2 e hyl bu y a e 2.95E- 02 0.2 ±0.1 0.10 ± 0.03 0.86 ± 0.05 0.19 ± 0.01 – 0.23 ± 0.04 0.1 ±0.1 0.3 ± 0.1 0.20 ± 0.03 – 0.2 ±0.1 e hyl isobu y a e 3.07E- 03 n.d. 0.3 ±0.2 0.30 ± 0.05 0.32 ± 0.01 0.34 ± 0.15 0.5 ±0.2 0.21 ± 0.09 0.5 ± 0.1 0.32 ± 0.06 0.38 ± 0.02 0.4 ±0.1 e hyl hexanoa e 3.09E- 01 2.7 ±0.8 4 ±2 7.0 ±0.6 6 ±5 – 5.9 ±2.3 3 ±1 4 ±2 6.7 ±0.9 – 3 ±5 e hyl oc anoa e 4.89E- 01 10 ±5 7 ±4 11 ±2 11 ±8 – 11 ±4 5 ±1 7 ±3 14 ±4 – 6 ±9 e hyl decanoa e 1.23E- 04 0.32 ± 0.05 2.2 ±0.7 8 ±2 4 ±2 – 4 ±1 2.9 ±0.9 3.6 ± 0.9 6.3 ±0.1 – 5 ±1 n.d. indica es ha he compound was no de ec ed o below de ec ion limi s. – indica es ha da a is no a ailable. M. Dena e al. Food Chemis y: X 9 (2021) 100116 5 hexanol and minima le els o mos e hyl es e s. BDX p oduced maxima le els o isobu anol and isoamyl alcohol. D80 p oduced maxima le els o ace ic, isobu y ic and decanoic acids. Finally, PERSY p oduced maxima le els o e hyl oc anoa e, oc anoic acids, e hyl lac a e and minima le els o isoamyl alcohol and isobu anol. In gene al, di e ences in oduced by he s ains we e o li le o mode a e magni ude. Le els o isoamyl alcohol anged om 160 o 310 mg/L, a ac o 2; hose o isobu anol om 20 o 60 mg/L, a ac o 3. These di e ences a e, howe e , la ge enough o ha e senso y signi i- cance (De-la-Fuen e-Blanco, S´ aenz-Na ajas, & Fe ei a, 2017). Di e - ences in he le els o ace ic acid we e much highe and amoun ed o a ac o 20, om jus 30 mg/L (IONYS) o 600 mg/L (RHONE). Howe e , lea ing aside IONYS, di e ences become mo e modes , anging om 340 o 600 mg/L, less han a ac o 2. Simila anges o a ia ion we e obse ed o hexanoic, oc anoic, decanoic and iso ale ic acids, while le els o isobu y ic acid anged a ac o close o 4. Le els o es e s we e, in gene al, e y low as a possible consequence o hei s ong e apo a- ion, and in some cases, hey we e no e en de ec ed. Lea ing aside IONYS, hei anges o a ia ion we e no la ge. 3.4. T ace a oma compounds. Va ie al o e men a i e o igin? The comple e da a se s wi h he concen a ions o up o hi y- ou ace a oma componen s in ecen ly e men ed and in aged wines, in he di e en con ols in oduced in he s udy, and he esul s o he di e en ANOVA s udies ca ied ou on he da a a e compiled in he Supplemen a y ma e ial (Tables S2–S6). Rega ding he a ie al o e men a i e o igin o he a oma com- pounds, he s udy o he con ols including o no he PAF ma e ial ex ac ed om he g apes and hose o he s including o no e men a ion (all compiled in Table S3), e eals ha some a oma compounds canno be unequi ocally classi ied in o e men a i e o a ie al. Ra he , he e a e se e al in e media e ca ego ies, as he answe o he ollowing h ee simple ques ions asked o each a oma compound, e eals; 1. Is he a oma compound p esen in e men ed con ols no con aining g ape PAF? 2. Is i p esen in un e men ed con ols con aining g ape PAF? 3. Is i a signi ican ly la ge amoun s in e men ed samples con aining g ape PAF han in he co esponding con ols no con aining g ape PAF? A posi i e answe o he i s ques ion implies a unequi ocal e men a i e o igin; he compound can be o med by yeas s om he basic lis o nu ien s supplied. On he con a y, a nega i e answe in- dica es ha he o ma ion o he compound equi es he p esence o g ape componen s. The answe s o he wo ollowing ques ions will indica e whe he he compound is p esen in he g ape PAF as speci ic p ecu so (posi i e answe o 2) and whe he yeas is equi ed o i s o ma ion (posi i e answe o 3). A ending o he answe s, i e di e en o igin- ela ed ca ego ies eme ged, as is schema ized in Fig. 1. 1) Pu e e men a i e compounds (answe s YNN) a e hose which we e p esen in e men ed samples no con aining PAF and whose le els we e no in luenced by he p esence o PAF. Compounds in his ca ego y we e isobu yl ace a e, e hyl iso ale a e, e hyl 2-me hylbu- y a e and γ-decalac one. 2) PAF-modula ed e men a i e compounds (answe s YNY) a e hose a oma compounds o med by yeas , bu whose le els a e signi i- can ly in luenced by he p esence o PAF. Compounds in his ca e- go y we e β-phenyle hyl ace a e, e hyl iso ale a e, e hyl leuca e, γ-oc alac one, β-ci onellol, ge aniol, ne ol. 3) Fe men a i e and a ie al a oma compounds (answe s YYY and YYN), a e hose a oma compounds which can be o med by yeas om basic nu ien s and which can be also ound in un e men ed con ols con aining PAF. Linalool and linalool oxide belong o his ca ego y. 4) Yeas -induced a ie al a oma compounds (answe s NNY), which a e hose a oma compounds ound exclusi ely in e men ed PAF. E hyl dihyd ocinnama e and β-ionone belong o his ca ego y. 5) Va ie al a oma compounds (answe s NYY), which a e hose com- pounds ound exclusi ely in samples con aining g ape PAF. Mos compounds belong o his ca ego y (massoia lac one, β-dam- ascenone, TDN, i ispi ane, Riesling ace al, anillin, ace o anillone, sy ingaldehyde, sy ingol, guaiacol, 4-e hyguaiacol, 4-e hylphenol, 4- inylguaiacol, 4- inylphenol, eugenol, me hoxyeugenol, ans-iso- eugenol, p-p opylguaiacol). I should be no ed ha , di ec ly o indi ec ly, le els o all compounds we e in luenced by he exis ence o e men a ion, which sugges s, as i will be u he seen in he nex sec ions, ha yeas is going o play a ele an ole on nea ly he comple e wine a oma p o ile. 3.5. Yeas s ain and aging: global o e iew Bo h ac o s, yeas s ain and aging ha e a s ong in luence on he ace a oma composi ion o wines, al hough aging is he dominan ac- o . Such dominance is mos e iden in he PCA plane gi en in Fig. 2, which shows he p ojec ion o he 60 samples (10 yeas s ×2 imes o analysis ×3 eplica es) in he wo i s dimensions. The i s componen (55.1% o he o iginal a iance) sepa a es samples a ending o age, wi h young samples on he le , and aged samples on he igh . The a iable loadings (no shown) e eals ha mos compounds, including no isop enoids, es e s, ola ile phenols, anillin de i a i es, inc ease du ing aging and ha only e penes (excep o linalool oxide), massoia lac one and phenyle hyl ace a e dec ease du ing aging. The plo e eals wo o he impo an cha ac e is ics. Fi s , ha samples e men ed wi h IONYS a e e y well sepa a ed om he o he s be o e and a e aging. Young samples because o i s highes le els in β-phenyle hyl ace a e, linalool and ge aniol, and aged samples because o i s highes le els o isobu yl ace a e, γ-oc a and decalac one. The second ema kable cha ac e is ic, is ha lea ing aside IONYS, he yeas s ain is an ac i e g ouping ac o in ace a oma compounds only a e aging. 3.6. E ec s o yeas s ain on wine a oma In o de o analyse he in luence o yeas aking in o conside a ion he dominance o he aging ime, wo di e en hea maps we e gene a ed wi h a oma compounds signi ican ly a ec ed by yeas s. The i s is gi en in Fig. 3A and includes da a om majo and ace a oma compounds Fig. 1. The i e o igin- ela ed ca ego ies in which a oma compounds o he expe imen should be classi ied. M. Dena e al. Food Chemis y: X 9 (2021) 100116 6 measu ed in young wines. The second can be seen in Fig. 3B and includes only ace a oma compounds in aged wines (Fig. 3B). The esul s o he hie a chical clus e ings a e also displayed on he le pa o each plo . Fig. 3A con i ms he singula i y o IONYS in young wines and he appa en ly low di e si y exis en be ween he o he s ains. A e aging, howe e , a much clea s uc u e eme ges as can be seen in Fig. 3B, whe e yeas s ains can be classi ied in o h ee di e en clus e s. Clus e 1 is in eg a ed by a qui e homogeneous g oup o med by D254, HPS, RHONE and CLOS and wi h mo e dissimila i y, by D80. The second clus e is o med by PERSY, OKAY, 71B and wi h mo e dissimila i y, BDX. And inally, IONYS is he single componen o he mos di e en clus e 3. I can also be seen ha yeas s in clus e 1 p oduced highe le els o pu e e men a i e compounds such as isobu yl ace a e, e hyl isobu y a e, 2-me hylbu y a e and iso ale a e and eleased highe le els o no isop enoids (TDN, i ispi ane and Riesling ace al). Clus e 2 eleased in gene al smalle le els o ola iles, excep o some ola ile phenols, such as anillin and guaiacol. 3.7. Modula ion o a ie al a oma The e ec o yeas s ain on hose genuine a ie al a oma compounds na u ally p esen in un e men ed con ols con aining g ape PAF can be assessed wi h he help o he plo s gi en in Fig. 4. The plo s compa e le els o a oma compounds ound in e men ed aged samples wi h hose ob ained in he un e men ed aged con ols. These ep esen a ions acili a e he iden i ica ion o he gene al ole o e men a ion on he a e o hese a ie al a oma compounds and also o he speci ic ole played by he s ain o yeas . Compounds can be classi ied in o h ee di e en ca ego ies depending on he e ec o e men a ion. 1s ca ego y.- Posi i e e ec o e men a ion. This ca ego y includes i e a oma compounds whose le els in e men ed samples we e much abo e hose ound in he un e men ed con ols, sugges ing ha some o he speci ic p ecu so s o hese a oma compounds could be o med by he ac ion o yeas s. Compounds in his ca ego y a e ace o anillone, β-ionone ( his one is no shown in Fig. 4, since i was only ound in young samples), e hyl dihyd ocinnama e, p-p opylguaiacol, 4-e hylphenol, ans-isoeugenol ( hese ou a e no ep esen ed in Fig. 4 since hey we e no de ec ed in he un e men ed con ol) and eugenol. Only in he case o eugenol he e ec o yeas was signi ican . Le els o ace o a- nillone in aged e men ed samples a e 10 imes highe han hose ound in he un e men ed con ol, howe e , no di e ence was obse ed be- ween he yeas s. 2nd ca ego y.- No e ec o e men a ion. Compounds in his ca ego y ha e in common ha , in a e age, le els o e men ed samples a e no dissimila o hose o he un e men ed con ols. The e a e howe e s ong di e ences a ending o he di e en ial e ec in oduced by yeas . In he cases o Riesling ace al in Fig. 4A, and o me hoxyeugenol and β-damascenone in Fig. 4B, he e is no e ec o yeas , which sugges s, Fig. 2. PCA ca ied ou on concen a ions o ace a oma compounds in he PAF-con aining samples e men ed by 10 S. ce e isiae s ains, analysed a e e men a ion and a e accele a ed aging. M. Dena e al. Food Chemis y: X 9 (2021) 100116 7 ha a leas o Temp anillo, he le els o hese a ie al a oma com- pounds canno be modula ed by yeas . The case o β-damascenone, which is an a oma enhance (Pineau, Ba be, Van Leeuwen, & Dubou - dieu, 2007) and modules he ipeness-cha ac e o ui y pe cep ion (San-Juan, Fe ei a, Cacho, & Escude o, 2011), dese es special men ion. Le els o his odo an in ecen ly e men ed samples we e abo e hose measu ed in he co esponding un e men ed con ols (Supplemen a y ma e ial S2 and S5) and we e signi ican ly in luenced by he s ain o yeas . Samples e men ed wi h IONYS showed le els up o 4 imes highe han hose ound in hose made wi h OKAY. This sugges s ha yeas s canno change he long- e m le el o his a oma compound bu can accele a e i s o ma ion. The second subca ego y includes anillin (4C), me hoxyeugenol (4B), and 4- inylphenol (4D), o which he s ain o yeas exe s a mode a e and signi ican in luence, so ha di e ences o a ound a 50% be ween he minimum and he maximum a e obse ed. 4-Vinylguaiacol can be also classi ied wi hin his subca ego y bu wi h wo s ains, IONYS and OKAY, showing a clea ou lie o e p oduc i e cha ac e . 3 d ca ego y.- Nega i e e ec o e men a ion. This ca ego y in- cludes a oma compounds whose le els in e men ed samples we e below hose ound in he un e men ed con ols. The e a e s ong di e ences be ween compounds a ending o he e ec played by he s ain o yeas . Massoia lac one (Fig. 4B) is a pa icula case whose le els d op o nea ly ze o in all e men ed samples wi h no di e ence be ween s ains. Mas- soia lac one is an impo an ma ke o o e - ipeness and con ibu o o p une a oma, whose le els a e known o dec ease du ing e men a ion (Pons, Allamy, La igne, Dubou dieu, & Da ie , 2017). Ou esul s e eal ha such dec ease in ensi ies du ing aging, which sugges s ha e men a ion educes also he p ecu so s. I would be o in e es o see whe he such educ ion is equally e ec i e in g apes con aining highe le els o p ecu so s o his molecule. Compounds in he ca ego y o which he s ain o yeas in oduced signi ican di e ences we e, om less o mo e in ense he e ec o s ain: sy ingol, i ispi ane, sy ingal- dehyde, guaiacol and TDN. I is appa en ha he speci ic p ecu so s o hese compounds a e me abolized di e en ly by he di e en s ains. This can ha e s ong echnological ele ance since guaiacol and TDN can ake pa in ele an odou aul s, and sugges s ha selec ed s ains a e a po en ially e ec i e emedial ool. Guaiacol is an a oma compound con ibu ing o he cha ac e is ic oas y-woody no es o Temp anillo, bu i can be a se ious o -odou de eloped wi h ime in wines made wi h g apes exposed o smoke (Ris ic, Van De Huls , Capone, & Wilkinson, 2017). Resul s in Fig. 4 e eal ha yeas s wi hin he clus e 1 seem o me abolize he p ecu so a highe le els. In he case o RHONE, le els o guaiacol we e educed by almos a ac o 3 compa ing wi h he un e - men ed con ol. Powe ul educ ions linked o speci ic yeas s ains can be also obse ed o TDN, known esponsible o ke osene no es de el- oped in aged Riesling wines. Wi h a 2 μ g/L de ec ion h eshold (Sacks e al., 2012), his compound will su ely also con ibu e o unpleasan no es in aged ed wines. The igu e also e eals ha yeas s in clus e 2, no ably 71B can TDN educe le els by a ac o 3 wi h espec o he con ol o D80. A simila yeas -induced and i ispi ane-independen dec ease o TDN le els has been ecen ly obse ed o non-Saccha o- myces yeas (Oli ei a & Fe ei a, 2019) bu o he bes o ou knowledge, i has no been obse ed o Saccha omyces s ains. This abili y can ha e a no able senso y impo ance, since le els o TDN a e expec ed o in- c ease due o clima e change (Win e hal e & G¨ ok, 2013). Fig. 3. Hea maps ob ained om no malized da a o he main ola iles signi ican ly modula ed by yeas s in young (A) and aged (A) wines. Compounds a e de ailed in he sub igu e C. M. Dena e al. Food Chemis y: X 9 (2021) 100116 8 Many a ie al a oma compounds in Fig. 4 de i e om e ulic acid and i s glycosides ( anillin, ace o anillone, isoeugenol, eugenol and 4- inylguaiacol). The highe le els o hese a oma molecules measu ed in aged e men ed samples sugges ha yeas ans o ms e ulic acid glycosides in o he co esponding a oma glycosides and ha hose ans o ma ions a e s ain speci ic. Aged wines made wi h BDX ha e maxima con en s o anillin and ace o anillone, hose made wi h IONYS ha e maxima con en s o eugenol while hose made wi h OKAY ha e minima le els o anillin and maxima le els o 4- inylguaiacol. Some o hose speci ici ies seem o be sha ed by s ains in he same clus e . Yeas s in clus e 2 show highe le els o anillin, ace o anillone and 4- inyl- guaiacol han hose in clus e 1. 4-Vinylguaiacol and 4- inylphenol dese e a speci ic commen , since hei le els a e ex emely dependen o he yeas s ain in unaged samples, wi h ac o s a ound 10 be ween he minimum and maximum concen a ions. Because o hei eac i i y, di e ences be ween maxima and minima sh ink o ac o s 3 (case o 4- inylguaiacol) and 1.7 (case o 4- inylphenol). In bo h cases, maxima le els we e obse ed o IONYS, and minima le els o BDX and HPS. 3.8. Modula ion o o he ele an a oma molecules Linalool and ge aniol a e he wo mos impo an e penols o wine. In he p esen case, hese wo molecules we e ha dly de ec ed in he un e men ed con ols, sugges ing ha he g ape ma e ial did no ha e much p ecu so s. The wo compounds we e ound in he con ols no con aining g ape ex ac , so ha yeas s we e able o o m weak, o mode a e in he case o IONYS, amoun s o hese molecules. Fe men ed unaged samples con ained bo h molecules a he expec ed concen a ion anges o Temp anillo (<6 μ g/L) excep samples e men ed by IONYS, whose le els we e abo e 20 μ g/L, which sugges s ha his qui e unique yeas s ain will p oduce young wines wi h ma kedly di e en cha ac- e s. Du ing aging, le els o hese compounds and he o he e penes dec eased (<1 μ g/L), while hose o linalool oxide, i s oxida ion p oduc , inc eased. E hyl leuca e o e hyl 2-hyd oxy-4-me hyl ale a e, is a ema kable a oma compound iden i ied in aged wines (Campo, Cacho, & Fe ei a, 2006) and sugges ed o be key in he speci ic blackbe y a oma o Bo deaux ed wines (Falcao, Ly a, Da ie , & Ba be, 2012). Resul s om his pape ha e shown ha maxima le els a e ound in aged PAF- con aining e men ed samples and ha he yeas s ain exe s a signi i- can in luence, wi h le els ound in IONYS, BDX, D80, PERSY and RHONE wice hose ound in 71B o OKAY (Table S5). E hyl es e s o b anched acids a e he mos impo an ui y es e s in aged ed wines, con ibu ing conce edly o ui y a oma (De-la-Fuen e- Blanco, S´ aenz-Na ajas, Valen in, & Fe ei a, 2020) and a e slowly o med by es e i ica ion o he co esponding acids syn hesized du ing e men a ion h ough Eh lich pa hway (Swiege s, Ba owsky, Henschke, & P e o ius, 2005). The in luence o yeas becomes mos ob ious a e aging. Minima le els a e ound in 71B, PERSY (clus e 2), and maxima in D80 and D254 (clus e 1), wi h di e ences as la ge as ac o s be ween 4 and 6.5. Isoamyl and phenyle hyl ace a es a e ele an in he a oma o young wines, since hei le els quickly ade by hyd olysis o he es e s. The in luence o yeas s in hei le els is o e whelming. In he case o β-phenyle hyl ace a e, le els in young wines (Table 5, Supplemen a y ma e ial) ange om 0.1 mg/L (clus e 1) o 1.5 mg/L in clus e 3, wi h le els in samples om clus e 2 be ween 0,18 and 0.29 mg/L. Simila esul s we e al eady obse ed o he isoamyl ace a e and isop opyl ace a es e apo a ed du ing e men a ion (Table 2). 4. Conclusions Vola iles los by e apo a ion du ing e men a ion a e mos ly e men a i e compounds and no g ape- ela ed a oma compounds. Quan i a i ely, apou s a e majo ly composed o 2-me hylp opanal, isop opyl ace a e, isoamyl ace a e, e hyl p opanoa e, isobu anol and isoamyl alcohol. While he ac ion o alcohols los is e y low, ha o he aldehydes and es e s can be well abo e 90% o he o al ola ile Fig. 4. Concen a ions o a ie al a oma compounds in PAF-con aining aged e men ed samples acco ding o hei le els in he un e men ed con ol. M. Dena e al. Food Chemis y: X 9 (2021) 100116 9 p oduced. The s ong impac exe ed by he s ain o yeas on wine a oma composi ion becomes in many cases only e iden a e aging, since le els o e hyl es e s o b anched acids, o mos g ape- ela ed a oma compounds and o many mino yeas s-de i ed a oma compounds mos ly inc ease du ing aging. The 10 s ains can be classi ied in o h ee clus e s showing ma ked di e ences in e men a i e and a ie al a oma p o iles. The bounda ies be ween e men a i e and a ie al a oma com- pounds a e in many cases blu ed. Fi s , he s udy has con i med a e men a i e o igin o linalool and ge aniol, ound a high le els in samples e men ed by one o he s ains. Second, he p esence o poly- phenolic and a oma ic ac ions om g ape exe s a s ong in luence on yeas me abolism and, hi d, he s ains o yeas no only hyd olyze glycosidic p ecu so s, bu me abolize qui e di e en ly he p ecu so s o ele an a oma compounds, such as phenolic acids and no isop enoids. These cha ac e is ics ha e in e es ing p ac ical consequences on he po en ial o yeas s o con ol he wine a oma p o ile and, mos ema kably, some wine aging a ibu es. Resul s ha e shown ha he a es o accumula ion o β-damascenone a e s ain- ela ed, and ha some s ains may be speci ically used o mi iga e ele an aging- ela ed o -odou s, such as hose ela ed o guaiacol, massoia lac one o TDN. CRediT au ho ship con ibu ion s a emen Ma ie Dena : In es iga ion, Fo mal analysis, Da a cu a ion, W i ing - o iginal d a , Visualiza ion. Dolo es P´ e ez: In es iga ion, W i ing - e iew & edi ing. Jos´ e Ma ía He as: Concep ualiza ion, Resou ces, W i ing - e iew & edi ing. Ampa o Que ol: Supe ision, Me hodology, W i ing - e iew & edi ing. Vicen e Fe ei a: Supe ision, Valida ion, Me hodology, W i ing - e iew & edi ing, Visualiza ion. Decla a ion o Compe ing In e es The au ho s decla e ha hey ha e no known compe ing inancial in e es s o pe sonal ela ionships ha could ha e appea ed o in luence he wo k epo ed in his pape . Acknowledgemen s The p ojec leading o his applica ion has ecei ed unding om he Eu opean Union’s Ho izon 2020 esea ch and inno a ion p og amme unde he Ma ie Skłodowska-Cu ie g an ag eemen No 764364. Ana Escude o is acknowledged o he supply o Temp anillo g apes and Sa a Fe e o-Del-Teso o he p epa a ion o Temp anillo mis ela. The au- ho s decla e ha no con lic o in e es exis s. Appendix A. Supplemen a y da a Supplemen a y da a o his a icle can be ound online a h ps://doi. o g/10.1016/j. ochx.2021.100116. Re e ences Aleg e, Y., A ias-P´ e ez, I., He n´ andez-O e, P., & Fe ei a, V. (2020). De elopmen o a new s a egy o s udying he a oma po en ial o winemaking g apes h ough he accele a ed hyd olysis o phenolic and a oma ic ac ions (PAFs). Food Resea ch In e na ional, 127, 108728. h ps://doi.o g/10.1016/j. ood es.2019.108728. Campo, E., Cacho, J., & Fe ei a, V. (2006). Mul idimensional ch oma og aphic app oach applied o he iden i ica ion o no el a oma compounds in wine. 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