Research Article. Rubén del Barrio-Galán, * Alejandro Cáceres-Mella, Marcela Medel-Marabolí and Álvaro Peña-Neira. Abstract

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Reserh Artile Reeived: 11 July 214 Revised: 16 Septemer 214 Aepted rtile pulished: 25 Septemer 214 Pulished online in Wiley Online Lirry: 27 Otoer 214 (wileyonlinelirry.om) DOI 12/jsf.6932 Effet of seleted Shromyes erevisie yest strins nd different ging tehniques on the polyshride nd polyphenoli omposition nd sensoril hrteristis of Cernet Suvignon red wines Ruén del Brrio-Glán, Alejndro Cáeres-Mell, Mrel Medel-Mrolí nd Álvro Peñ-Neir Astrt BACKGROUND: The ojetive of this work ws to study the effet of two Shromyes erevisie yest strins with different pilities of polyshride liertion during loholi fermenttion in ddition to susequent ging on lees with or without ok wood hips s well s ging with ommeril intive dry yest on the physil, hemil nd sensoril hrteristis of Cernet Suvignon red wines. RESULTS: The HPS (high levels of polyshrides) yest strin relesed higher mounts of polyshrides (429 g L 1 )thn EC1118 (39 g L 1 ) during loholi fermenttion, ut the onentrtion equlized during the ging period (424 nd 417 g L 1 respetively). All ging tehniques inresed the polyshride onentrtion, ut the inrese ws dependent on the tehnique pplied. A higher liertion of polyshrides redued the onentrtion of most of the phenoli fmilies nlyzed. Moreover, no ler effet of the different ging tehniques used in this study on olor stiliztion ws found. The HPS wines were etter vlued thn the EC1118 wines y the pnel of tsters fter loholi fermenttion. CONCLUSION: In generl, the HPS wines showed etter physiohemil nd sensoril hrteristis thn the EC1118 wines. Aording to the results otined during the ging period, ll ging tehniques ontriuted to improve wine qulity, ut it ws diffiult to estlish the tehnique tht llowed the est wine to e otined, euse it depended on the ging tehnique used nd the period of ging. 214 Soiety of Chemil Industry Keywords: polyshrides; phenoli ompounds; red wines;shromyes erevisie yest strins; sensory qulity 2132 INTRODUCTION Reently, severl sientifi studies hve foused on testing the importnt role tht yest ell wll polyshrides from Shromyes erevisie ply in the tehnologil nd sensoril hrteristis of red wines. Certin studies ssert tht these ompounds n redue wine stringeny nd itterness, 1 4 improve olor stility 5 7 nd enhne mouth-feel nd persistene. 1,2,4,8,9 Aording to these studies, the most importnt polyshrides of enologil interest re mnnoproteins, whih re lierted into wines during loholi fermenttion 1 nd therefter during yest utolysis. 11,12 For this reson, severl vinifition yest strins of S. erevisie hve een ommerilized y mnufturers in the lst few yers with the ojetive of relesing greter mounts of mnnoproteins into wines during loholi fermenttion. Alterntively, for severl yers, winemkers hve used ging on lees in white wines fter loholi fermenttion to ontinue the liertion of polyshrides y utolyti proesses in order to improve wine omplexity. However, this tehnique ws most reently used in red wines. The relese of polyshrides during the ging of wines on lees is too slow owing to the required temperture nd ph onditions, whih re not the most suitle for this proess. 13 Therefore vriety of ommeril produts rih in yest ell wll polyshrides from S. erevisie hve reently een developed nd supplied y mnufturers to provide the sme enefits s the ging of wines on lees ut in shorter period of time. 1,8 Thus wineries n put the finl wines on the mrket in shorter period of time ut with similr qulity to the wines ged on lees. In terms of struture nd hemil omposition, these yest produts re very heterogeneous, whih is minly due to Correspondene to: Ruén del Brrio-Glán; Deprtment of Agro-Industry nd Enology, Fulty of Agronomil Sienes, University of Chile, Post Offie Box 14, Sntigo, Chile. E-mil: rdelrriogln@gmil.om Deprtment of Agro-Industry nd Enology, Fulty of Agronomil Sienes, University of Chile, Post Offie Box 14, Sntigo, Chile J Si Food Agri 215; 95: 2132 2144 www.soi.org 214 Soiety of Chemil Industry

Effet of yest strins nd ging methods on red wine qulity www.soi.org the different intivtion (therml or enzymti), extrtion nd purifition proesses used in their prodution. Moreover, they re lssified into four lrge groups: intive yests, utolyzed yests, yest wlls nd yest extrts. 14 It is lso well known tht ging wine in ok rrels imprts numerous enefits ut entils ost tht, in some ses nd given the tul glol wine mrket, nnot urrently e mintined. Therefore the ddition of ok hips is used to redue prodution osts nd to provide greter flexiility nd mngeility. Additionlly, these hips improve the sensory hrteristis of wines, suh s inresing their sweetness, struture nd mouth-feel, 15,16 whih is minly due to the ontriution of ok wood polyshrides nd hydrolyzed tnnins. In ddition, this tehnique n e omined with ging on lees, euse the polyshrides lierted from yest lees n pir with ok wood tnnins, reduing their stringeny nd itterness. 5 Beuse very few experimentl nd sientifi trils hve een onduted with these tehniques, they ll rise mny douts nd questions for winemkers nd tehniins out oth their use nd their finl effet on wines. In ddition, no sientifi studies hve een performed tht simultneously ompre the pility of different S. erevisie yest strins to relese lrge mounts of polyshrides into wines s well s the susequent pplition of the ging tehniques mentioned ove. Furthermore, the effets of these tehniques hve not een ssessed in ny sientifi or experimentl study on Chilen wines. However, it is importnt to note tht most studies onduted with these tehniques used shorter periods of ging. 1,8,17 Therefore it is neessry to pply longer ging periods to evlute their effet on wines. Thus the min ojetive of this work ws to study the effet of two S. erevisie yest strins with different polyshride liertion pilities during loholi fermenttion s well s susequent ging on lees nd other lterntive prties on the physil, hemil nd sensoril hrteristis of Cernet Suvignon red wines. Aording to the lst report issued y SAG, 18 the totl volume of wine produed in 213 ws 12.8 million hl, 34.6% of whih ws Cernet Suvignon. Thus it is importnt to pply these tehniques to the most importnt vriety ultivted for wine prodution in Chile. EXPERIMENTAL Winemking proess nd tretments The study ws rried out using Cernet Suvignon red grpes supplied y the Cliterr winery (Errzuriz group) loted in Colhgu vlley, Chile (34.63 S, 71.37 W). The grpes seleted for this study ( 65 kg) were hrvested ording to their tehnologil mturity, sed on optiml sugr ontent ( 24.8 Brix) nd totl idity ( 5.5 g L 1 H 2 SO 4 ), nd then trnsported to the winery. One there, the trditionl winemking proess for red wines ws followed. Briefly, fter de-stemming, rushing nd susequent sulfite ddition, the must ws trnsferred into two different 25 L stinless steel tnks for loholi fermenttion. One of these tnks ws inoulted with ommeril S. erevisie ynus yest strin (3 g hl 1 Llvin EC1118), whih is the onventionl yest used y the Cliterr winery. The other tnk ws inoulted with nother ommeril S. erevisie yest strin (3 g hl 1 Uvferm HPS), whih produes high levels of polyshrides during loholi fermenttion (2 3% higher thn other yests), ording to the mnufturer s speifitions. Both yest strins were supplied y Lllemnd-South Ameri (Sntigo, Chile). The wine fermenttion proess stritly followed the mnufturing tehniques of red wines mde from Cernet Suvignon in the Cliterr winery. Aloholi fermenttion ws rried out t ontrolled temperture (21 25 C); one fermenttion ws omplete, the wines were kept in the tnks for 5 dys to llow the sedimenttion of gross lees, then they were rked off gin. After this proess, 6 L of eh type of fermented wine ws trnsported to the pilot plnt of the Deprtment of Agro-Industry nd Enology, Fulty of Agronomil Sienes, University of Chile, Sntigo, Chile One there, the wines were kept in the tnks until mlolti fermenttion, whih hd egun spontneously in the winery, ws omplete. After tht, the sulfur dioxide level ws orreted to 35 μll 1 free SO 2, nd oth types of wine were distriuted into 25 L food-grde plsti tnks. The wine ws kept in these tnks for 5 dys to promote the sedimenttion of fine lees, whih were olleted for use in the tretments rried out with lees, s desried y Del Brrio-Glán et l. 1 The different tretments were rried out in triplite nd lsted 4 months. The tretments were: ontrol wines (the wines otined fter loholi nd mlolti fermenttion without ny tretment) (C); wines ged on lees (olleted fine lees, 3 ml L 1 ) (L); wines ged on lees (olleted fine lees, 3 ml L 1 )ndfrenhok(querus petre) wood hips (3 g L 1 medium-tosted Noile Sweet L oenologie du ois; Lffort, Frne) (L + CH); nd wines with ommeril intive dry yest (CIDY) dded (3 g hl 1 Opti-LEES). The wood hips used hd length etween 7 nd 2 mm nd produed sweetness senstion ording to the supplier, nd the CIDY ws rih in low-moleulr-weight polyshrides, minly mnnoproteins, ording to the supplier (Lllemnd-South Ameri). All wines were resuspended nd homogenized with the lees, hips nd CIDY through two tonnges per week for the first 2 months. Therefter, only one tonnge ws performed per week for the lst 2 months to prevent wine oxidtion nd miroiologil ltertion. Chemil regents Methyl ellulose (15 P visosity t 2 g L 1 ), etldehyde, glli id, prototehui id, ffei id, syringi id, p-oumri id, feruli id, ellgi id, ftri id, tyrosol, tryptophol, queretin, myrietin, stilin, (+)-tehin, ( )-epitehin, mlvidin-3-gluoside, dextrn nd petin stndrds were purhsed from Sigm-Aldrih Chemil Co. (St Louis, MO, USA). Polyethylene memrnes (.45 nd.22 μm pore size) were quired from EMD Millipore (Billeri, MA, USA). Sodium sulfte (nhydrous), potssium metisulfite, vnillin (99%), ethyl ette, diethyl ether, sodium hydroxide, eti id, formi id, sulfuri id, ethnol, hydrohlori id nd high-performne liquid hromtogrphy (HPLC)-grde etonitrile, methnol nd mmonium formte were purhsed from Merk (Drmstdt, Germny). All regents were of nlytil grde or higher. Enologil prmeter nlyses Totl (TA) nd voltile (VA) idity, ph (S22 SevenCompt ph/ion, Mettler Toledo, Sntigo, Chile), SO 2 F (sulfur dioxide free) nd SO 2 T (sulfur dioxide totl) nd loholi degree (A ) were evluted following the OIV offiil nlytil methods. 19 Spetrophotometri nlyses Totl polyphenols were determined y UV sorne t 28nm nd expressed s mg L 1 glli id. 2 Totl nthoynins were mesured t 52 nm nd expressed s mg L 1 mlvidin-3-gluoside. 21 Totl tnnins, expressed s g L 1 J Si Food Agri 215; 95: 2132 2144 214 Soiety of Chemil Industry wileyonlinelirry.om/jsf 2133

www.soi.org R del Brrio-Glán et l. 2134 (+)-tehin, were evluted fter their preipittion with methyl ellulose nd mesured t 28 nm. 22 Color intensity ws evluted using the method desried y Glories, 23 nd CIEL prmeters (with illuminnt D65 nd 1 oserver onditions) were determined ording to Pérez-Mgriño nd González-Snjosé. 24 The perentges of opigmented nd polymeri olor were evluted using the method desried y Hermosín-Gutiérrez. 25 All mesurements were performed on UV/VIS 17 Phrmspe spetrophotometer (Shimdzu, Kyoto, Jpn). HPLC nlyses Anthoynin ompounds were nlyzed y tking 2 ml of wine nd filtering it through.22 μm pore size memrne, then 15 μl liquots of the smple were sujeted to reverse phse hromtogrphi seprtion t 2 C using Chromolith C18 olumn Merk (Drmstdt, Germny). The photodiode rry detetor (DAD) ws set to monitor from 21 to 6 nm. The two moile phses used were (A) wter/formi id (9:1 v/v) nd (B) etonitrile. A grdient ws pplied t flow rte of 1.1 ml min 1 from to 22 min nd 1.5 ml min 1 from 22 to 35 min s follows: 96 85% A from to 22 min, 85 15% A from 12 to 22 min nd 85 7% A from 22 to 35 min. Quntifition ws performed y pek re mesurement t 52 nm. Anthoynins were quntified nd expressed s mg L 1 mlvidin-3-gluoside. Clirtion urves t 52 nm were otined y injeting different volumes of stndrd solutions under the sme onditions used for the smples. 26 Low-moleulr-weight phenoli ompounds (non-flvonoids nd flvonoids) were nlyzed using the method desried y Peñ-Neir et l. 27 A 5 ml liquot of red wine ws extrted with diethyl ether (3 2 ml) nd ethyl ette (3 2 ml) to onentrte the phenoli ompounds. The orgni frtions were omined, dehydrted with 2.5 g of nhydrous sodium sulfte nd evported to dryness under vuum t 3 C. The solid residue ws dissolved in 2 ml of methnol/wter (1:1 v/v) nd filtered through.22 μm pore size memrne. Aliquots (25 μl) of the finl solution were sujeted to reverse phse hromtogrphi seprtion. Compounds were nlyzed with n HPLC 11 Series system (Agilent Tehnologies, Snt Clr, CA, USA) onsisting of G1315B DAD, G1311A quternry pump, G1379A degsser nd G1329A utosmpler. A reverse phse Nov-Pk C18 olumn (4 μm, 3.9 mm i.d. 3 mm; Wters (Milford, MA, USA)) ws used for the HPLC-DAD nlysis of individul phenoli ompounds t 2 C. Eh mjor pek in the hromtogrms of extrts ws hrterized y oth retention time nd sorption spetrum (from 21 to 36 nm). The quisition time ws 1 s. Clirtion urves t 28 nm were produed y injeting stndrd solutions efore extrtion under the sme onditions s the smples nlyzed over the rnge of onentrtions oserved (r 2.93). Queretin glyosides, dimeri proynidins, glltes nd stilene gluoside, for whih no stndrds were ville, were quntified using stndrd urves for queretin, (+)-tehin, glli id nd trns-resvertrol respetively. All qulittive nd quntittive nlyses of phenoli omposition (inluding their extrtion step) were performed in triplite. Polyshrides were nlyzed y high-performne size exlusion hromtogrphy with refrtive index detetion (HPSEC-RID) in order to determine their moleulr distriution nd onentrtion. HPSEC-RID ws performed using n Agilent 126 Infinity Series liquid hromtogrph (Agilent Tehnologies, Snt Clr, CA, USA) equipped with G1362A refrtive index detetor, G1311B quternry pump, G1316A olumn oven with two Shodex olumns, OHpk SB-83 HQ nd SB-84 HQ, onneted in series (3 mm 8 mm i.d.; Show Denko, Tokyo, Jpn), nd G1329A utosmpler. Polyshride frtions were quntified using lirtion urves produed with dextrns nd petins (Leuonosto mesenteroides). 26 Sensory evlution A desriptive nlysis of the wines ws performed y trined sensory pnel of 12 people who were ll workers nd students t the Deprtment of Agro-Industry nd Enology. The wines were evluted in individul temperture-ontrolled tsting ooths, nd wter nd unslted rkers were provided for plte lensing. Using ompletely rndomized order, liquots (2 ml) of wine were served t 18 19 C in drk wine-tsting glsses (RCristl, Mendoz, Argentin) leled with three-digit ode. Drk wine-tsting glsses were used to prevent the interferene of visul senstions nd to fous the ttention of pnelists on gusttive senstions. Between eh smple, pnelists hewed on rker nd then rinsed their mouths with wter. An unstrutured liner 15 m sle (where = sene of senstion nd 15 = extremely high senstion ) ws used to evlute eight sensory ttriutes (idity, sweetness, lohol, itterness, red fruits, stringeny, persistene, nd mouth-feel). All nlyses were rried out fter mlolti fermenttion (AMLF) nd fter 2 nd 4 months of tretment (2MT nd 4MT respetively). Sttistil nlysis Anlysis of vrine (ANOVA) nd Tukey s honestly signifint differene (HSD) test were employed for men seprtion, using signifine level of 95% (P <.5). To otin summrized nd synthesized informtion from the lrge set of vriles nd to etter understnd the effet of the different tretments pplied, multivrite sttistil tehnique, ftor nlysis, ws utilized. All sttistil nlyses were onduted using Sttgrphis Centurion Version 15.2 (SttPoint Tehnologies, In., Wrrenton, VA, USA) nd Exel 27 Version 12. (Mirosoft Corp., Redmond, WA, USA). RESULTS AND DISCUSSION Effet on enologil prmeters The enologil prmeters were nlyzed to study the effet of the different ging tehniques ssyed on these prmeters (Tle 1). No sttistilly signifint differenes were deteted etween the wines fermented with the EC1118 nd HPS yests. Additionlly, no sttistilly signifint differenes were found etween the different ging tretments pplied. These results greed with similr pulished studies tht used these tehniques with other grpe vrieties. 1,8,28 Effet on polyshride ontent Four frtions of polyshrides were identified, quntified nd lssified ording to their verge moleulr weights: frtion I, >2 kd; frtion II, 2 3 kd; frtion III, 6 8 kd; frtion IV, 1 kd. As seen in Fig. 1, frtions IV nd III showed the highest polyshride onentrtion in the EC1118 nd HPS wines respetively. The wines fermented with the HPS yest strin presented, fter the MLF period, higher onentrtion of low-moleulr-weight polyshrides, whih orresponded with frtions III nd IV, nd higher totl polyshride onentrtion thn the wines fermented with EC1118. The polyshride onentrtion inresed during the ging period in the treted wileyonlinelirry.om/jsf 214 Soiety of Chemil Industry J Si Food Agri 215; 95: 2132 2144

Effet of yest strins nd ging methods on red wine qulity www.soi.org Tle 1. Enologil prmeters (TA, g L 1 H 2 SO 4 ;VA,gL 1 eti id) in Cernet Suvignon red wines AMLF 2MT 4MT EC1118 HPS EC1118 HPS EC1118 HPS Prmeter C C C L L + CH CIDY C L L + CH CIDY C L L + CH CIDY C L L + CH CIDY 13.5 ± 13.6 ± 14.5 ± 13.6 ± 13.3 ± 13.4 ± 14.4 ± 13.4 ± 13.6 ± 13.6 ± 13.6 ± 13.6 ± 13.3 ± 13.3 ± 13.3 ± 13.3 ± 13.7 ±.2 A ns 13.3 ± 3.62 ± 3.7 ±.75 ± 11.6 ± 1.84 31. ± 2.33 3.66 ± 3.5 ±.5.75 ± 15.3 ± 1.78 33.9 ± 1.59 3.7 ± 3.1 ±.75 ± 14.2 ± 1.61 35.8 ± 1.48 3.66 ± 3.18 ±.6.7 ±.5 12.7 ± 1.38 31. ± 2.69 3.62 ± 3.9 ±.8 ± 14. ± 2.65 32.1 ± 2.72 3.67 ± 3.14 ±.5.8 ±.5 1. ± 1.76 29.7 ± 2.13 3.64 ± 3.1 ±.75 ± 14.3 ± 1.63 33.4 ± 1.45 3.65 ± 3.1 ±.75 ± 11. ± 1.34 34.2 ± 2.79 3.69 ± 3. ±.6.4 ± 24. ± 2.4 71. ± 1.73 3.7 ± 3.5 ±.6.39 ± 25.3 ± 1.52 7.7 ± 2.19 3.71 ± 3.5 ±.5.37 ± 24.3 ± 1.58 72.8 ± 1.8 3.7 ± 3.8 ±.5.39 ± 22.3 ± 2.8 71.3 ± 3.51 3.65 ±.1 3.1 ±.4 ± 24. ± 2. 71.7 ± 2.74 3.66 ± 3.15 ±.4 ± 3.65 ± 3.12 ±.39 ± 3.65 ± 3.1 ±.4 ± 3.66 ±.5 3.25 ±.35 ± ph ns 3.66 ±.6 TA ns 3.3 ±.7 VA ns.37 ± 24. ± 1. 24.3 ± 1.53 SO 2 F ns.. 23. ± 1.73 71.7 ± 1.53 73.3 ± 1.93 67.7 ± 4.87 4.4 ± 5 SO 2 T ns 5.5 ±.7 Vlues re expressed s men ± stndrd devition (n = 3). AMLF, fter mlolti fermenttion; 2MT, 2 months of tretment; 4MT, 4 months of tretment. No sttistilly signifint differenes. s well s the ontrol wines, minly fter the 2MT period. The oserved inrese in the polyshride onentrtion of the ontrol wines hs een explined y other uthors, 1 who stted tht these ompounds remin in the wine in olloidl stte linked to other ompounds or tht they re the result of utolysis of the remining ded yest present in the wine. After the 4MT ging period, this onentrtion ws mintined or deresed, ut the hnge ws dependent on the ging tretment pplied. However, fter the entire ging period, oth types of fermented wine (EC1118 nd HPS) showed similr onentrtion of totl polyshrides. This suggested tht the HPS yest llowed fster liertion of yest polyshrides, minly mnnoproteins, during loholi fermenttion thn the EC1118 yest, ut fter the 2MT ging period, nd minly fter 4MT, oth types of wine were similr. Moreover, ll ging tehniques used (L, L + CH nd CIDY) generlly inresed, in oth EC1118 nd HPS wines, the totl polyshride onentrtion with respet to the ontrol wines, ut these results depended on the different moleulr weight frtions nlyzed nd the ging tehnique used. These differenes were more evident in the EC1118 wines thn in the HPS wines. The results otined gree with other studies performed with similr ging tehniques. 1,3,8,17,29 31 Plomero et l. 29 studied the ging on lees of different strins of yest over 142 dys, ut in model medium, nd oserved progressive inrese in the onentrtion of polyshrides. Gudlupe nd o-workers 3,3,31 evluted the use of mnnoprotein-overproduing yest strin nd the ddition of ommeril mnnoproteins to red wines during or fter loholi fermenttion. They noted n inrese in or mintenne of the neutrl (mnnoproteins) nd totl polyshrides during rrel nd ottle ging, lthough these effets were dependent on the dose nd produt used. Del Brrio-Glán et l. 1,8 evluted the ging of wines on lees, hips nd severl ommeril yest derivtive produts rih in polyshrides (mnnoproteins) nd showed tht ll tehniques studied llowed the relese of higher onentrtions of totl nd neutrl polyshrides to the wine, lthough the type nd ontent of these ompounds depended on the tehnique used nd the yest derivtive dded, sine eh produt hd different purity nd omposition. Similr results were otined y other uthors 17 using n over-lees ging tehnology pplied to red wines. Effet on phenoli ompounds nlyzed y spetrophotometri methods Tle 2 shows the totl polyphenols (TP), totl tnnins (TT) nd totl nthoynins (TACY) nlyzed in the red wines using different spetrophotometri methods. Severl sttistilly signifint differenes were identified. For exmple, fter the MLF period, the wines fermented with HPS showed sttistilly signifintly lower ontent of TP, TT nd TACY thn the wines fermented with EC1118. As expeted, the ontents of the different phenoli ompounds nlyzed deresed s the ging period progressed nd different results were found s funtion of the tretment used nd the ging period nlyzed. In ft, the HPS wines generlly presented lower TP ontent thn the EC1118 wines over ll ging periods. In ddition, ll wines treted with the different ging tehniques (L, L + CH nd CIDY) presented lower TP ontent thn the ontrol wines for oth HPS nd EC1118 wines, with the wines treted with L presenting the lowest vlues. However, for TT, no sttistilly signifint differenes were found etween the HPS nd EC1118 wines. The treted wines presented lower TT ontent thn the ontrol wines only fter the 2MT ging period. For TACY, the HPS wines showed lower ontent thn the EC1118 wines fter the J Si Food Agri 215; 95: 2132 2144 214 Soiety of Chemil Industry wileyonlinelirry.om/jsf 2135

www.soi.org R del Brrio-Glán et l. 2136 mg L 1 mg L 1 mg L 1 mg L 1 5 d d d d 45 4 35 3 25 2 15 1 mg L 1 5 6 5 4 3 2 1 12 1 8 6 4 2 18 16 14 12 1 8 6 4 2 25 2 15 1 5 A C B D E AMLF e de d d d 2MT d d d d 4MT EC C EC L EC L+CH EC DL HPS C HPS L HPS L+CH HPS DL e de dd d d e e e Figure 1. Conentrtions of (A) totl polyshrides nd (B E) polyshride frtions I IV respetively in wines. AMLF, fter mlolti fermenttion; 2MT, 2 months of tretment; 4MT, 4 months of tretment. For eh period (AMLF, 2MT or 4MT), different letters indite sttistilly signifint differenes (P <.5) etween vlues. 2MT ging period, nd this ontent ws lower in ll treted wines ompred with the ontrol. However, different results were found fter 4MT; while the EC1118 treted wines presented lower ontent thn the ontrols, the HPS treted wines exhiited opposite results, with higher TACY ontent eing oserved. These results gree with those otined y other uthors who used tehniques similr to those mentioned ove over 2 months. 1,8 The lower ontents of the different phenoli ompounds oserved for the HPS nd EC1118 wines nd the differene etween the different treted wines with respet to the ontrols ould e explined y higher liertion of polyshrides y the HPS yest ompred with the EC1118 yest fter loholi fermenttion. The greter mount of these polyshrides lierted y the different ging tehniques, ompred with the ontrol wines, ould intert with the different phenoli ompounds, reduing their onentrtion in wines. These results gree with those otined y other uthors, whih reveled the pility of the mnnoprotein-overproduing yest strin, yest lees nd severl ommeril produts rih in mnnoproteins to retin or dsor different wine phenoli ompounds. 3,8,3 33 The smller differenes found etween the two types of fermented wine fter the 4MT ging period for ertin ompounds were most likely due to the ontent of polyshrides in the EC1118 wines, whih ontinued to inrese, rehing similr vlues to the HPS wines. However, these results depended on the tretment used nd the ompound nlyzed. Effet on phenoli ompounds nlyzed y HPLC methods The monomeri nthoynins identified nd quntified in this study were grouped into the gluosylted nthoynins (delphinidin-3-gluoside, ynidin-3-gluoside, petunidin-3- gluoside, peonidin-3-gluoside nd mlvidin-3-gluoside), the etylted nthoynins (delphinidin-3-(6-etyl)-gluoside, ynidin-3-(6-etyl)-gluoside, petunidin-3-(6-etyl)-gluoside, peonidin-3-(6-etyl)-gluoside nd mlvidin-3-(6-etyl)-gluoside) nd the oumroylted nthoynins (delphinidin-3-(6-poumroyl)-gluoside, ynidin-3-(6-p-oumroyl)-gluoside, petunidin-3-(6-p-oumroyl)-gluoside, peonidin-3-(6-p-oumroyl)- gluoside nd mlvidin-3-(6-p-oumroyl)-gluoside). As seen in Tle 2, the group of gluosylted nthoynins presented the highest onentrtion. The TACY ontent, nlyzed y spetrophotometri methods, nd the ontent of monomeri nthoynins deresed during the ging period, most likely owing to the intertion of these ompounds with other phenoli ompounds, suh s tnnins or other nthoynins, y polymeriztion nd opigmenttion retions s explined y severl uthors, 34 37 whih n form more omplex moleulr strutures. The wines fermented with HPS yest ontined lower monomeri nthoynin onentrtions thn those fermented with EC1118 yest. The most signifint differenes were found in mlvidin-3-gluoside, mlvidin-3-(6-etyl)-gluoside nd mlvidin-3-(6-p-oumroyl)-gluoside (dt not shown), whih were the ompounds with higher onentrtions in oth types of wine. As disussed ove, the polyshrides (minly mnnoproteins) lierted from the yest ell wlls n intert or dsor these phenoli ompounds. This ould led to the formtion of more omplex strutures tht n remin stle in wine s olloids or preipitte owing to their higher moleulr weight. These results oppose those otined y some uthors, 3,31 euse no sttistilly signifint differenes in the monomeri nthoynin ontent were found etween the wileyonlinelirry.om/jsf 214 Soiety of Chemil Industry J Si Food Agri 215; 95: 2132 2144

Effet of yest strins nd ging methods on red wine qulity www.soi.org Tle 2. Color prmeters (CI, olor intensity; L, C, H, nd, CIEL prmeters; CC, opigmentted olor; PC, polymeri olor), totl polyphenols (TP, mg L 1 glli id), totl tnnins (TT, g L 1 tehin), totlnthoynins(tacy,mgl 1 mlvidin-3-gluoside) nd monomeri nthoynins (mg L 1 mlvidin-3-gluoside) in Cernet Suvignon red wines AMLF 2MT 4MT EC1118 HPS EC1118 HPS EC1118 HPS Prmeter C C C L L + CH CIDY C L L + CH CIDY C L L + CH CIDY C L L + CH CIDY CI 11.4 9.13 14.1f 13. d 13.1d 13.5e 12.8 12.1 12.2 11.8 16.3 g 16.ef 15.9e 16.2 fg 13.d 11.4 11.8 12.3 L 49.8 56.9 44.5 45.8 45.5 44.7 46.7 48.3d 48.1d 49.2e 37.8 38.2 37.7 38. 46.3 5.5e 49.2d 47.9 C 52. 46.1 57.1 55.4 55.5 56.4 54.4e 53.1 53.1 52.9d 54.f 53.1e 52. 54.5 g 52.1 d 49.4 5.5 52.3d H 6.3 8.6 9. 8.5 9.4 9.4 11.8e 11.4d 12.2f 11.3d 16.1 d 15.6 14.4 16.2d 16. d 15. 14.7 14.7 51.7 45.6 56.4e 54.8 54.8 55.7d 53.3 52.1 51.9 51.9 51.9f 51.2e 5.3 d 52.3 g 5 47.7 48.9 5.6d 5.7 6.9 8.9 8.2 9. 9.2 11.1e 1.5d 11.2e 1.4d 15. 14.3 12.9 15.3 14.4 12.8 12.8 13.3 %CC 33.2 3.2 32.4 3.9 3.2 34.4d 25.6 25.6 25.6 25.6 23.9 23.4 23.1 24.1d 28.4 g 28.1f 26.7e 28.5 g %PC 23.7 25.6 26.2 26.5 28.1 26.7 37.9d 37.4d 37.6d 36.4 5.7e 52.4f 56.2 g 5.8e 43.2 46.6d 44.1 45.4 TP 1612 1416 1588 g 1552e 1559f 1592 g 1418 147 1429d 1397 1519f 1465d 1478e 1524f 141 1344 1385 1382 TT 1.4 1.21 1.14.88.973.959 1.13.95 1.2.924 1.2.772 1.2 1. 1.6.91 1.1.813 TACY 647 567 653 h 615f 67e 627 g 515d 489 476 54 283 27 253 276 349d 385f 372e 37e Monomeri nthoynins Gluosylted ACY 531 418 44e 471f 385d 39d 359 336 317 363 14de 16 59 97 123 d 162ef 149e 179f Aetylted ACY 156 136 151f 131e 115d 11d 92. 83. 86.9 58.3 49.5f 4.8de 3.8 27.3 36.3 d 24.7 37.6de 42.7e Coumroylted ACY 39.2 34.6 34.2d 31.4 39.2e 3.8 3 23.7 21.2 27.2 28.7d 22.5 13.2 14.4 21.7 1.5 11.8 12.4 TMACY 727 589 623e 634e 539d 531d 481 443 425 451 218ef 169 13 139 181 d 197de 198de 234 g J Si Food Agri 215; 95: 2132 2144 214 Soiety of Chemil Industry wileyonlinelirry.om/jsf For eh period (AMLF, 2MT or 4MT), different letters indite sttistilly signifint differenes (P <.5) etween vlues within row. AMLF, fter mlolti fermenttion; 2MT, 2 months of tretment; 4MT, 4 months of tretment; TMACY, totl monomeri nthoynins. 2137

www.soi.org R del Brrio-Glán et l. 2138 wines fermented with the mnnoprotein-overproduing yest strin nd those fermented with the use of ommeril mnnoproteins during loholi fermenttion. In generl, during the ging period, ll HPS wines showed lower monomeri nthoynin onentrtions thn the EC1118 wines. As ourred in the AMLF period, the most signifint differenes were found in mlvidin-3-gluoside, mlvidin-3-(6-etyl)-gluoside nd mlvidin-3-(6-p-oumroyl)-gluoside (dt not shown). Furthermore, oth the HPS nd EC1118 treted wines presented lower onentrtions of these ompounds fter the 2MT period, with some exeptions, euse this effet ws different depending on the tretment nd the ompound nlyzed. Thus ll treted wines presented lower onentrtions of etylted nthoynins thn the ontrol wines, with the wines treted with CIDY showing the lowest ontent. Similr results were otined for the glyosylted nthoynins, ut the HPS wines treted with L showed higher ontent thn the ontrol nd the other treted wines, nd the EC1118 wines treted with CIDY showed similr ontent to the ontrol wines. Finlly, ll treted wines presented lower oumroylted nthoynin ontents thn the ontrol wines in this period, with the exeption of the EC1118 wines treted with L + CH, whih presented higher ontent. After the 4MT ging period, the results otined for the totl monomeri nthoynin (TMACY) ontent, nd prtiulrly for the glyosylted nthoynins, orrelted well with those otined for TACY y spetrophotometri methods. Moreover, ll EC1118 treted wines showed lower TMACY ontents thn the ontrol wines, while the opposite ws otined for ll HPS treted wines, with exeption of the wines treted with L, whih were lower in TMACY. However, oth EC1118 nd HPS treted wines showed lower ontents of oumroylted nthoynins thn the ontrol wines. As with TACY, these results gree with those otined y other uthors. 1,8 Tle 3 shows the low-moleulr-weight phenoli ompounds identified nd quntified in the wines. These ompounds were lso grouped into phenoli fmilies to more lerly understnd oth the results nd the sttistilly signifint differenes otined. Tyrosol, glli id nd (+)-tehin were the ompounds present in the highest onentrtions. After the MLF period, the wines fermented with HPS yest hd lower ontents of ll phenoli fmilies thn the EC1118 wines, with the exeption of hydroxyinnmi ids (HCA), hydroxyinnmi id trtri esters (HCATE) nd flvonol glyosides. As explined previously, the lower onentrtion of these ompounds in the HPS wines ould e due to higher intertion nd/or dsorption etween the mnnoproteins nd/or the polyshrides relesed in higher onentrtion thn with the EC1118 yest during loholi fermenttion. Severl sttistilly signifint differenes were lso found during the ging period, ut these differenes depended on the tretment pplied, the ging period studied nd the type of phenoli ompounds nlyzed. For hydroxyenzoi ids (HBA), the most signifint differenes were found t the end of the ging period, where ll HPS wines hd lower HBA ontents thn EC1118 wines. In oth ses the treted wines hd lower HBA ontents thn the ontrol wines, with the exeption of the HPS wines treted with L + CH tht were higher in HBA, most likely owing to the liertion of glli nd ellgi ids from the ok wood hips, s explined y the uthors of omprle previous study. 38 Similr results were found for HCATE, minly in the EC1118 wines. These results were lso shown y severl uthors who used ging on lees, hips nd ommeril mnnoproteins in red wines. 1,8,3,39,4 For HCA, the most importnt differenes etween the HPS nd EC1118 wines were found fter the 2MT ging period, with the onentrtion eing higher in the ltter. The onentrtion of HCA inresed during the ging period of the wines, whih ws explined y severl uthors to e result of the enzymti hydrolysis of their esterified forms or the hydrolysis of oumroylted nthoynins. 41 43 Severl uthors lso noted tht some HCA suh s ffei, p-oumri nd feruli ids n inrese in wines ged in ok wood y hydroloholysis; 38,44 however, under the onditions used in the present study, this effet not ws oserved. Finlly, the onentrtions of HCA nd HCATE re generlly well orrelted during the ging period. A similr trend in the onentrtion of stilenes nd phenoli lohols ws found during the ging period, whih inresed t the end of this period. The HPS wines hd lower ontent of oth types of these phenoli ompounds thn the EC1118 wines, ut ertin differenes were found etween the ging tretments. After the 2MT period, ll treted wines hd lower stilene ontent thn the ontrol wines, with the wines treted with L + CH, CIDY nd L showing lower vlues in the order speified. Similr results were found fter the 4MT ging period, ut the order of the lower vlues ws L + CH, L nd CIDY. However, for the phenoli lohols, the EC1118 treted wines hd higher onentrtion thn the ontrol wines, nd no signifint differenes etween the EC1118 nd HPS treted wines were found fter the 2MT ging period. After the 4MT period, ll treted wines hd lower ontent thn the ontrol wines for oth HPS nd EC1118 wines, ut it ws dependent on the tretment used. This lower phenoli lohol ontent grees with tht otined in previous studies rried out with the sme or similr ging tehniques. 39,4 Moreover, similr trend ws oserved in the totl flvnol monomers ((+)-tehin nd ( )-epitehin) nd the totl proynidins (B3, B4 nd B2); s seen in Tle 3, the onentrtion deresed during the ging period. After the 2MT period, the onentrtion of totl flvonol monomers ws signifintly lower in the HPS wines thn in the EC1118 wines, ut smller differene etween the treted nd ontrol wines ws found. Only the EC1118 wines treted with CIDY hd signifintly lower flvnol onentrtion thn the ontrol wines. After the 4MT period, fewer signifint differenes were found etween the wines developed with oth yest strins, nd only the HPS ontrol wines hd lower onentrtion thn the EC1118 ontrols. In ontrst, the HPS wines treted with CIDY presented higher onentrtion thn the sme wines fermented with EC1118 yest. However, the EC1118 nd HPS wines treted with L + CH nd CIDY showed lower ontents of totl proynidins thn the ontrol wines fter the 2MT period. Severl uthors hve shown tht the intertion etween ommeril mnnoproteins from S. erevisie, mnnoprotein-overproduing yest strin (during or fter loholi fermenttion) nd yest lees with proynidins resulted in signifintly lower ontent in wines treted with these produts. 3,31,39,4 No signifint differenes fter the 4MT ging period were found etween the HPS nd EC1118 wines, nd only proynidin B4 ws deteted. As expeted, the flvonol glyosides nd the glyones showed similr trend to HCA nd HCATE during the ging period. The onentrtion of flvonol glyosides deresed nd tht of glyones inresed, minly owing to the hydrolysis of glyosides, whih llowed the liertion of glyones into wines. After the 2MT period, ll HPS wines hd lower onentrtion of flvonol glyosides thn the EC1118 wines, with the exeption of the wines treted with L. The EC1118 wines treted with L hd higher glyone ontents thn the respetive HPS wines. The pplied tretments seemed to ffet the EC1118 wines more thn the HPS wines for these phenoli ompounds. Thus ll treted wines presented lower ontents of oth glyoside nd glyone forms, with wileyonlinelirry.om/jsf 214 Soiety of Chemil Industry J Si Food Agri 215; 95: 2132 2144

Effet of yest strins nd ging methods on red wine qulity www.soi.org Tle 3. Low-moleulr-weight phenoli ompounds quntified (mg L 1 ) in Cernet Suvignon red wines AMLF 2MT 4MT EC1118 HPS EC1118 HPS EC1118 HPS Compound C C C L L + CH CIDY C L L + CH CIDY C L L + CH CIDY C L L + CH CIDY Non-flvonoid phenoli ompounds Glli id 16.3 12.1 18.4 17.7 19.8 18.3 15.6 13.7 19. 13.7 23.7f 17.5de 18.1e 16.d 12.3 9.6 11.6 1.5 Prototehui id.81.747 1.31.926.781.983 2.24 2.11 1.66 2.21 2.25 1.62 1.65 1.78 2.11 1.84 1.65 1.92 Methyl gllte.718.559.819.797.751.819.89.89.679.755.772.653.93.832.825.867.891.84 Vnilli id.855.84.835.763.87.823.789.789.781 1.3 1.7.923 1.18.883.962 1.1 1.74.975 Syringi id 1.7 1.1 1.74 1.54 1.75 1.76 1.71 1.71 2.18 2.3 1.74 1.37 1.46 1.24 1.3 1.17 1.35 1.27 Ethyl gllte 3.38 2.46 2.12 1.65.97 2.3 2.4 2.46 1.35 1.52 2.38 2.36 2.19 2.1 1.71 1.46 1.63 1.73 Ellgi id 5.1 4.49 5.11 3.95 4.68 5.1 4.3 5.67 6.13 4.47 2.3 2.47 3.1 1.63 2.69 3.5 4.39 3.27 Totl HBA 28.2 22.4 3.4 27.3 29.5 29.8 27.5 28.2 31.8 25.7 34.f 26.9de 28.5e 24.5 d 21.9 19. 22.6 2.4 t-cffei id 2.84 2.99 1.98 2.27 2.26 2.13 3.71 3.71 3.5 2.87 4.58 4.44 4.42 3.77 4.21 4.64 4.35 4.7 t-p-coumri id 2.3 2.29 2.61 2.49 2.33 2.78 4.3 4.32 3.8 3.57 5.26 5.16 4.98 4.99 5.29 4.73 5.12 5.77 t-feruli id.743.72 ND ND ND ND ND ND ND ND 1.45.736.71.88 ND ND ND ND Totl HCA 5.88 5.99 4.59 4.76 4.59 4.9 8.1 8.84d 6.85 6.44 9.48 9.61 9.4 8.77 9.5 9.36 9.47 1.5 t-cftri id 1.84 1.92 1.87 1.45 1.51 1.54 2.1 2.1 1.76 2.22 2.2d 1.78 1.52 1.6 1.57 1.37 1.39 1.56 -Coutri id.625.722 1.4 1.7.96 1.6.935 1.7.722.75.824.748.82.643.749.72.744.753 t-coutri id.784.687.7.678.451.621.781.862.514.535 1.17 1.14.913.864.91.775.825.851 t-fertri id.26.276.426.288.289.294.576.68.727.551 ND ND ND ND ND ND ND ND.231.214 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND t-p-coumri id hexose ester Totl HCATE 3.73 3.82 4.39 3.48 3.21 3.51 4.4 4.75 3.72 4.6 4.1d 3.67 3.24 3.11 3.23 2.86 2.96 3.17 t-resvertrol.434.392.77d.653 d.569.543.513.569.283.311.253.292.284.22.235.256.237.212-3-gluoside -Resvertrol.377.329.369.221 45.223.363.34.292.31.461.331.255.273.363.39.292.261-3-gluoside t-resvertrol ND ND.656d.593.443.466.61 d.447.317.368.67d.52.387.438.354.38.293.323 -Resvertrol 42 48.37.268 59 88.25.227 5.215.81 d.646.66.667.91d.55.512.63 Totl stilenes.953.869 2.4d 1.74 1.32 1.42 1.74 1.58 1.4 1.2 2.12e 1.79d 1.53d 1.6 1.72 d 1.31 1.26 1.42 Tyrosol 25.7 2. 28.6 37.1 35.3 33.1 2.3 2.2 17.2 19.3 4.7e 33.9 d 31.3d 34.3d 31.6d 26.2 27.7 24.1 Tryptophol 2.97 2.28 3.74 3.32 3.2 4.15 2.97 2.6 2.54 3.18.66.512.686.313.783.765.924.748 28.7 22.3 32.3 4.4 38.3 37.2 23.2 22.8 19.7 22.5 41.4d 34.4 32. 34.6 32.4 26.9 28.6 24.8 Totl phenoli lohols Flvonoid phenoli ompounds Ctehin 17.1 1.6 1.6de 11.5e 1.2de 9.93d 7.21 7.85 5.39 6.1 6.15e 5.16 d 3.87 4.3 4.82 5.64de 4.77 5.71de Epitehin 5.81 4.37 9.82d 7.38 8.41 d 5.85 4.84 6.11 5.7 6. 4.65 3.42 2.47 2.53 2.74 3.51 2.51 2.97 22.9 15. 2.4d 18.9 d 18.7 d 15.8 12.1 14. 1.5 12. 1.8e 8.58 d 6.34 6.83 7.56 9.15 d 7.28 8.68d Totl flvnol monomers Proynidin B3 1.17 1.2.657d.392 ND. 54.273.369 d 63.293d ND ND ND ND ND ND ND ND Proynidin B4.99.98.555d.262 44.362 ND ND ND ND 1.6.924.987.894 1.7 1.38 1.22 1.1 Proynidin B2 3.97 3.86 3.44d 3.39d 3.6d 3.25 d 2.25 2.6 1.36 1.38 ND ND ND ND ND ND ND ND J Si Food Agri 215; 95: 2132 2144 214 Soiety of Chemil Industry wileyonlinelirry.om/jsf 2139

www.soi.org R del Brrio-Glán et l. 214 Tle 3. Continued AMLF 2MT 4MT EC1118 HPS EC1118 HPS EC1118 HPS Compound C C C L L + CH CIDY C L L + CH CIDY C L L + CH CIDY C L L + CH CIDY Totl proynidins 6.13 5.78 4.65e 4.4de 3.74 d 3.76 d 2.52 2.96 1.53 1.68 1.6.92.99.89 1.7 1.38 1.22 1.1 Queretin 5.51 4.91 5.85de 5.26 d 4.44 5.12 6.3e 5.46de 4.35 4.82 7.55d 6.89 d 5.29 5.85 6.44 6.56 5.85 7.6 d Myrietin 4.83 4.88 5.28d 4.3 3.74 4.4 5.87d 5.61 d 4.1 4.71 6.59 5.81 4.72 5.67 6.7 6.12 5.61 6.67 Astilin ND ND ND ND ND ND ND ND ND ND 2.13 2.1 2.5 1.92 1.64 2.24 1.92 2.38 Totl flvonol 1.3 9.79 11.1 9.56 8.18 9.51 11.9 11.1 8.36 9.52 16.3 14.8 12.1 13.4 14.1 14.9 13.4 16.1 glyones Flvonol glyosides 14.2 14. 16.9 13.7 11.4 13.2 12.1 1. 11.6 11.5 13.2 12. 1.5 1.6 11. 12.1 11.9 13.2 For eh period (AMLF, 2MT or 4MT), different letters indite sttistilly signifint differenes (P <.5) etween vlues within row. ND, not deteted. AMLF, fter mlolti fermenttion; 2MT, 2 months of tretment; 4MT, 4 months of tretment; HBA, hydroxyenzoi ids; HCA, hydroxyinnmi ids; HCATE, hydroxyinnmi id trtri esters. the wines treted with L + CH showing the lowest ontent. Fewer differenes were found in HPS wines, nd only those treted with L + CH nd CIDY showed lower ontents of glyones thn the ontrol wines nd those treted with L. No sttistilly signifint differenes were found in the flvonol glyosides, nd these results gree with those otined y other uthors. 39 Fewer differenes etween the EC1118 nd HPS wines were found fter the 4MT ging period. Moreover, the HPS wines treted with CIDY showed higher onentrtion of glyosides nd glyones thn the respetive EC1118 treted wines. On the other hnd, different effet ws lso found etween the ging tretments fter this period. In generl, ll EC1118 treted wines hd lower onentrtions of oth types of flvonols thn the ontrol wines, with the exeption of the wines treted with L, whih mintined similr ontents. Some uthors noted tht these ompound n t s opigments euse of their higher ility to intert with nthoynins, 45 47 whih ould explin the lower flvonol ontent in these tretments. This effet n lso e explined y the intertion of flvonols with the polyshrides lierted during these ging tretments. However, in HPS wines, only those treted with CIDY presented higher ontent thn the ontrols, while the rest of the treted wines mintined onentrtion similr to the ontrols. These results greed with those otined y some uthors. 39,4 Effet on olor of wines Tle 2 shows severl olor prmeters evluted to oserve possile differenes in the olor evolution of the wines fermented with the two yest strins nd the susequent evolution during ging on lees nd the other elerted ging tehniques used in this study. The wines fermented with EC1118 yest presented higher olor intensity (CI) thn the wines fermented with HPS yest fter the MLF period nd the ging period. The HPS wines showed higher vlues of the L (lightness), H (hue) nd (hromti oordinte of lue/yellow olors) CIEL prmeters thn the EC1118 wines fter the MLF period. These results were mintined during the ging period, with some exeptions, euse fter the 4MT ging period, some EC1118 treted wines showed H nd vlues similr to or higher thn those of the HPS wines. As explined in the literture, 48 the ging of wines entils n inrese in the L, H nd prmeters, nd there is n inverse reltionship etween these prmeters nd CI. Therefore the results otined for these prmeters, s shown in Tle 2, re well orrelted fter the MLF period nd during the ging period, with the exeption of the H nd prmeters fter 4MT. This trend ws mintined for oth EC1118 nd HPS treted wines, euse, in generl, ll treted wines presented lower CI nd higher L thn the ontrol wines, with the exeption of the EC1118 treted wines, whih hd similr L to the ontrol wines. However, ll treted wines generlly presented lower vlues of H nd thn the ontrol wines. These results indite tht the higher liertion of polyshrides y the HPS yest nd the different ging tretments did not produe olor stiliztion, ut olor loss my hve ourred owing to polymeriztion of the polyshrides with the phenoli ompounds responsile for the olor of wines. The trends of the perentges of opigmented olor (%CC) nd polymeri olor (%PC) re typilly opposite during the ging proess, euse opigmenttion retions our more in young wines while polymeri retions usully our in ging wines. These vlues re well orrelted euse, while %CC deresed during ging, %PC inresed. Furthermore, the EC1118 wines showed higher %CC vlues thn the HPS wines fter the MLF period, nd opposite results were found for the %PC vlues. These wileyonlinelirry.om/jsf 214 Soiety of Chemil Industry J Si Food Agri 215; 95: 2132 2144

Effet of yest strins nd ging methods on red wine qulity www.soi.org results were mintined fter the 2MT ging period ut were opposite fter 4MT. This trend n e explined euse, s noted y some uthors, 49 the retions tht form opigmented omplexes our very esily nd re reversile over time, ut the omplexes lso disssoite esily nd re unstle. Fewer signifint differenes in %CC etween tretments were found during the ging period, nd only the EC1118 wines treted with CIDY showed higher vlues thn the ontrol nd the remining treted wines. In ontrst, ll treted wines presented higher vlues of %PC thn the ontrol wines fter the 4MT ging period, with the exeption of the EC1118 wines treted with CIDY, whih mintined similr ontent to the ontrol wines. Higher %PC ws oserved in the EC1118 wines thn in the HPS wines fter the ging period, whih ould explined y higher ontriution of these treted wines to olor stiliztion. More polymeriztion retions ould our etween the nthoynins nd other phenoli ompounds, whih ould ontriute to olor stiliztion. In ddition, the %PC vlues re well orrelted with TACY, TMACY nd CI. However, the higher vlues of %PC oserved in oth EC1118 nd HPS treted wines (with respet to the ontrols) were not well orrelted with the CI vlues, euse the expeted hypothesis ws tht higher %PC vlues ontriuted to the inrese in CI vlues, whih would explin the improvement in olor stiliztion. Severl uthors suggest tht polyshrides nd mnnoproteins n intert with nthoynins nd tnnins, thus preventing their preipittion nd improving their olor stility. 5 8,5 Menwhile, other uthors 4,51 indite tht polyshrides n t s olor stilizers or enhne the preipittion of phenoli polymers to use loss of olor, depending on their moleulr weight, dsorent hrter, hrge nd struture. However, other reent studies did not find n improvement in wine olor intensity nd olor stility using mnnoproteins. 1,3,31 Multivrite nlysis Ftor nlysis ws pplied on polyshride, polyphenoli nd olor vlues to study the ssoition of different vriles. To minimize the numer of vriles nd etter understnd the results otined, only the grouped vlues were used for the low-moleulr-weight phenoli ompounds nd monomeri nthoynins. The vrimx rottion riterion ws pplied nd only ftors with eigenvlues greter thn unity were seleted. All dt were stndrdized efore the ftor nlysis. The ftor nlysis seleted five ftors with n eigenvlue greter thn 1, whih explined 91.2% of the totl vrine. Tle 4 shows the ftor lodings for eh vrile of the seleted ftors. The vriles with higher loding vlues ontriute most signifintly to the explntory mening of the ftors. As seen in Tle 4, most vriles used for this multivrite nlysis were ssoited with ftor 1, whih explined 51.8% of the totl vrine. Figure 2 shows the distriution of the different wines studied in the plne defined y ftors 1 nd 2, whih explined 68.6% of the totl vrine. These ftors llowed the seprtion of oth types of fermented wine (EC1118 nd HPS) during the ging period s well s the seprtion of the wines treted with the different ging tehniques ssyed. This effet ws most evident t the end of the ging period (4MT). Sensory nlysis The sensory nlysis ws foused on the gusttive ttriutes (Fig. 3) desried ove. Vrious sttistilly signifint differenes were found in the AMLF, 2MT nd 4MT periods, depending on the Tle 4. Ftor lodings fter vrimx rottion of treted wines Item Ftor 1 Ftor 2 Ftor 3 Ftor 4 Ftor 5 Eigenvlue 15. 4.86 2.96 2.4 1.21 Cumultive vrine 51.8 68.6 78.8 87.1 91.2 Totl polyphenols.568.784 Totl tnnins.969 Totl nthoynins.969 L.339.919 C.757.565 H.963.857.446.931 % opigmented olor.258.252.862.296 % polymeri olor.959 Color intensity.296.928 Polyshride.755.418 frtion I Polyshride.87.27 frtion II Polyshride.754.46.324 frtion III Polyshride.721.284.572 frtion IV Totl polyshrides.36.848 Totl monomeri.986 nthoynins Gluosylted.969 nthoynins Aetylted.968 nthoynins Coumrylted.815.29 nthoynins Totl hydroxyenzoi.47.557.483 ids Totl.95 hydroxyinnmi ids Totl.767 hydroxyinnmi id trtri esters Totl monomers.955 Totl proynidins.946 Flvonol glyosides.515.655 Totl flvonol glyones.722.599 Totl stilenes.475.719 Totl lohols.767.435 Lodings lower thn n solute vlue of ±.25 re not shown. Bold numers indite the higher loding vlues tht ontriute to eh ftor. yest strin used, the ging tretment used nd the period nlyzed. Moreover, the wines fermented with the HPS yest strin showed lower vlues of idity, itterness nd stringeny nd higher vlues of red fruits nd persistene thn the wines fermented with the EC1118 yest. Conversely, no signifint differenes were found etween the EC1118 nd HPS wines during the entire ging period, ut some differenes were found etween the ging tretments nd the ontrol. Thus the tster pnel indited tht, in generl, ertin treted wines presented higher vlues of red fruits, persistene nd mouth-feel thn the ontrol wines J Si Food Agri 215; 95: 2132 2144 214 Soiety of Chemil Industry wileyonlinelirry.om/jsf 2141

www.soi.org R del Brrio-Glán et l. 2 1,5 1 A ACIDITY 12 1 MOUTH-FEEL 8 SWEETNESS Ftor 2,5 4MT -1,5-1 -,5,5 1 1,5 2 2,5 -,5 2MT -1 PERSISTENCE 6 4 2 ALCOHOL -1,5 2142-2 Ftor 1 EC C EC L EC L+CH EC CIDY HPS C HPS L HPS L+CH HPS CIDY Figure 2. Distriution of different wines defined y ftor 1 nd ftor 2. 2MT, 2 months of tretment; 4MT, 4 months of tretment. fter the 2MT ging period, ut sttistilly signifint differenes were only found in few ses. For exmple, only the HPS wines treted with L + CH presented higher vlues of red fruits thn the ontrol wines. In ddition, the EC1118 nd HPS wines treted with L shower higher persistene nd lower stringeny thn the ontrol wines. Furthermore, oth EC1118 nd HPS wines treted with L + CH nd CIDY showed higher vlues of mouth-feel thn the ontrol wines. A similr trend ws oserved fter the 4MT ging period, where ll treted wines showed lower vlues of itterness nd stringeny nd higher persistene nd mouth-feel thn the ontrol wines, ut signifint differenes were not found in ll ses. Additionlly, the EC1118 nd HPS wines treted with L showed signifintly lower vlues of itterness thn the ontrol wines, nd the EC111 wines treted with L nd CIDY s well s the HPS wines treted with L presented lower vlues of stringeny. The EC1118 wines treted with L + CH nd CIDY nd the HPS wines treted with L presented higher vlues of persistene thn the ontrol wines. Finlly, only the EC1118 wines treted with L + CH showed higher vlues of mouth-feel thn the ontrol wines. Del Brrio-Glán et l. 1 lso studied the effet of ging on lees, hips nd ommeril yest derivtives rih in mnnoproteins on severl gusttive ttriutes of red wines; in generl, they disovered lower vlues of stringeny nd green tnnins nd higher vlues of grssy, lne nd overll puntution thn in the ontrol wines. In nother study rried out y Del Brrio-Glán et l. 8 regrding the effet of six different ommeril yest derivtives rih in mnnproteins nd with different purifition degrees of red wines, they noted tht ll treted wines presented lower vlues of green tnnins thn the ontrol wines. Aording to their explntion, these types of tnnins produe negtive senstions, inluding intense stringent nd idi senstions with strong green or hereous notes. Other uthors note tht the intertions etween these produts nd the tnnins n inrese roundness nd softness on the plte. 2,4,5 Therefore the use of these ging tehniques n improve the gusttive senstions of red wines, likely owing to the inresed liertion of polyshrides nd/or mnnoproteins into the wines. CONCLUSIONS The use of HPS yest llowed quiker relese of higher mounts of polyshrides (minly mnnoproteins) with low moleulr weights during loholi fermenttion thn the EC1118 yest. B ASTRINGENCY PERSISTENCE C MOUTH-FEEL ASTRINGENCY PERSISTENCE MOUTH-FEEL ASTRINGENCY RED FRUITS ACIDIT Y 12 1 8 6 4 2 RED FRUITS ACIDITY 12 1 8 6 4 2 BITTERNESS SWEETNESS BITTERNESS SWEETNESS BITTERNESS ALCOHOL ALCOHOL RED FRUITS EC C EC L EC L+CH EC CIDY HPS C HPS L HPS L+CH HPS CIDY Figure 3. Sensory nlysis grphis of different wines fter (A) mlolti fermenttion, (B) 2 months of tretment nd (C) 4 months of tretment. Sttistilly signifint differenes (P < ). This onentrtion generlly equlized during the ging period, inditing tht the EC1118 yest relesed these polyshrides more slowly. The different ssyed ging tehniques inresed the polyshride onentrtion ut depended on the tehnique pplied. However, these ging tehniques seemed to hve more of n effet on the EC1118 wines. wileyonlinelirry.om/jsf 214 Soiety of Chemil Industry J Si Food Agri 215; 95: 2132 2144