Tangential-flow membrane clarification of Malvar (Vitis vinifera L.) wine: incidence on chemical composition and sensorial expression

Tangen ial- low memb ane cla i ica ion o Mal a (Vi is ini e a L.) wine: incidence on chemical composi ion and senso ial exp ession Ma in P odano 1*, Ma ga i a Azna 2, Juan M. Cabellos3, Visi ación Vacas3, F ancisco López4, Ma ía T. He nández5and Ma ía I. Es ella5 1Ins i u o de In es igación en Ciencias de la Alimen ación (CIAL) (CEI, CSIC-UAM), Calle Nicolás Cab e a, 9, Ciudad Uni e si a ia de Can oblanco, 28049 Mad id, Spain 2Depa amen o de Química Analí ica, Uni e sidad de Za agoza, Campus Rio Eb o, Ma ía de Luna 3, 50018 Za agoza, Spain 3Ins i u o Mad ileño de In es igación y Desa ollo Ru al, Ag a io y Alimen a io (IMIDRA), C a. A-2, km 38.2, inca “El Encín”, 28800 Alcalá de Hena es (Mad id), Spain 4Depa amen d’Enginye ia Química, Facul a d’Enologia, Uni e si a Ro i a i Vi gili, A . Paisos Ca alans, 26, 43007 Ta agona, Spain 5Ins i u o de Ciencias y Tecnología de Alimen os y Nu ición (ICTAN, CSIC), Calle Juan de la Cie a, 3, 28006 Mad id, Spain *Co esponding au ho ([email p o ec ed]) Mal a whi e wine (Vi is ini e a L.) was cold se led (CSW) and cla i ied by angen ial- low memb ane il a ion (TFMF). A 500 kDa molecula mass cu -o memb ane was used. Fil a ion lux o 49-48 L/hm2was achie ed a ansmemb ane p essu e o 0.7 ba . The ea men p oduced a comple ely cla i ied wine wi h u bidi y o 0.11 NTU, bu also a 10.3 % loss o p o eins, which could be ela ed o he dec ease o some la ou compounds. The CSW and he memb ane il e ed wines (MFW) we e assessed by means o hei a oma and phenolic composi ion, as well as hei senso y p ope ies. The esul s showed ha he gene al physicochemical pa ame e s and mos o he analysed phenolic compounds we e no o sligh ly (up o 7.6 %) a ec ed by he TFMF p ocess. Ne e heless, he ea men p oduced an impo an loss o some key a oma compounds: up o 43 % o a y acid and alcohol es e s and up o 26 % o highe alcohols. Mos a ec ed we e a oma species wi h highe molecula masses and lowe pola i ies. Senso y analysis con i med he global dec ease in wine a oma. TFMF ea men p oduced also an inc ease o 52 % o he wine benzaldehyde con en . Mal a wine, cla i ica ion, angen ial- low il a ion, ola ile composi ion, phenol composi ion, ch oma og aphy BSA: bo ine se um albumin, CSW: cold se led wine, ΔNTU: ne u bididy a e hea es , HPLC: high pe o mance liquid ch oma og aphy, log P: oc anol-wa e pa i ion coe icien , Lp: memb ane hyd aulic pe meabili y, MFW: memb ane il e ed wine, NTU: nephelome ic u bidi y uni s, OAV: odou ac i i y alue, Q : eed low, OT: odou h eshold, PS: polysul one, PTM: ansmemb ane p essu e, PVDF: poly inylidene luo ide, TFMF: angen ial low memb ane il a ion, TPI: o al phenol index ABSTRACT KEYWORDS ABBREVIATIONS Recei ed: 29 Ap il 2010 y Accep ed: 13 No embe 2019 y Published: 21 No embe 2019 DOI:10.20870/oeno-one.2019.53.4.2480 VINE AND WINE OPEN ACCESS JOURNAL 725 OENO One 2019, 4, 725-739 © 2019 In e na ional Vi icul u e and Enology Socie y - IVES INTRODUCTION Cla i ica ion is an essen ial uni ope a ion o winemaking. I imp o es no only appea ance o wines, bu also is e y impo an o hei physicochemical and mic obial s abili y. This s a emen is especially heigh ened o whi e wines, as hey ha e o be clea , wi h u bidi y lowe han 1 NTU (Ribé eau-Gayon e al., 2006a). Pos - e men a ion cla i ica ion is usually ca ied ou a a ious s ages, which can include na u al o o ced se ling, inning and/o il a ion. As na u al se ling can ex end oo long in ime, o ced se ling (cold o cen i ugal) is p e e ed o indus ial p oduc ion. Al e na i ely, ining wi h ben oni es, p o eins (egg whi es, gela ine, caseina es, e c.), silica gel o annins con ibu es also o speed up he se ling ea men (Ribé eau-Gayon e al., 2006b; Li a e al., 2015). In mos o he cases, cla i ica ion ends up wi h a s age o il a ion. The classic ea men includes a combined il a ion wi h cellulose-based il e pla es o dia omaceous ea hs and ine polishing and mic obial s abiliza ion o he wine wi h 0.45 o 1.2 µm po e sizes plea ed memb ane il e s (Hidalgo Togo es, 2002). In all cases, hese il e aids a e use ul un il hey clog, which means ha hei use gene a es was es and subsequen ly, en i onmen al p oblems. An al e na i e o hese ea men s wi h a g owing accep ance in he winemaking indus y is he angen ial- low (TFMF) o c oss- low memb ane il a ion. In his case, he gene a ed u bulen wine eed low d ags insoluble pa icles om he memb ane su ace and e u ns hem back o he eed essel, a oiding hei deposi ion on he memb ane su ace and/o po es. In he p ac ice, some ine pa icles and/o colloidal ma e ial build up a ine gel laye on he memb ane su ace (concen a ion pola iza ion) (Bake , 2004) ha p oduces a subsequen dec ease o il a ion lux and memb ane ouling (El Rayess e al., 2011). Ne e heless, memb anes employed in angen ial- low il a ion a e use ul du ing long pe iods (mon hs and yea s) hanks o he eco e y o he memb ane pe meabili y a e chemical cleaning (P odano e al., 2013). Nowadays, memb ane manu ac u e s o e a wide ange o memb ane ma e ials (o ganic polyme and mine al sin e ed laye s), designs (spi al wound, hollow ib e o capilla y, mul ichannel, e c.) and po e sizes, which allow many op ions o choice o any conc e e applica ion. Howe e , his di e si y also p oduces some con usion o he use s because o he speci ici y o each applica ion and he di icul ies o p edic hei e ec . In mos cases, o cla i ica ion o whi e wines, p oduce s o e hyd ophilic memb ane ma e ials (cellulose es e s, poly inylidene luo ide (PVDF), polysul one (PS), polye he sul one, e c.), compac memb ane designs (hollow ib e o spi al wound) and memb anes wi h po e sizes be ween 0.2 and 5 µm (El Rayess e al., 2011). Fo whi e wine cla i ica ion, some ul a il a ion memb anes a e also a ailable. In gene al, memb anes wi h smalle po e sizes (0.1 o 0.22 µm) gi e be e cla i ica ion e ec s ( u bidi ies < 0.5 NTU) and mo e s able il a ion luxes, as he sizes o hei po es a e conside ably smalle han he sizes o he mos abundan pa icles in wines and g ape-de i ed p oduc s (Se ano e al., 1992; Ribé eau-Gayon e al., 2006a; P odano e al., 2013). The p oblem o such ine cla i ica ion is ha an impo an pa o he colloidal ma e can be emo ed also (Ulb ich e al., 2009; El Rayess e al., 2011; Obe hols e e al., 2013), leading o dec ease o wine la ou in ensi y. I is no ewo hy o men ion ha mos bibliog aphic s udies dealing wi h CFMF conside only senso y o gene al physicochemical e alua ion o he cla i ica ion (Feuilla e al., 1987; Flo es e al., 1991; Se ano e al., 1992; Bu on e al., 2014) o p oblems ela ed o memb ane s abili y: ouling, oulan s o memb ane ma e ials (Ve nhe e al., 1999; Ribé eau-Gayon e al., 2006a). S udies ela ed o he e ec o TFMF on wine la ou componen concen a ions and he in ensi y o hei pe cep ion a e sca ce and in he cases whe e hey we e a ailable, da a abou he TFMF p ocess we e no a ailable (Se ano e al. 1984, Flo es e al., 1991; Se ano and Pae zold, 1994). Tha is why he main aim o his s udy was o e alua e he junc ion o p ocess pa ame e s, a oma and phenolic composi ion and senso y e alua ion o a young whi e wine be o e and a e ine cla i ica ion wi h one o he memb anes wi h igh es po es a ailable on he ma ke , he 500 kDa molecula mass cu -o Romicon memb ane. Ano he objec i e o his s udy was he chemical cha ac e iza ion o a wine, p oceeding om he li le known c . Mal a (Vi is ini e a L.) g apes (VIVC 7254). This is a whi e a ie y, which p oduces ui y wines (San os e al., 2004). I s cul i a ion is admi ed only in o he denomina ions o o igin ‘Vinos de Mad id’ and ‘La Mancha’. Due o he sca ce in o ma ion Ma in P odano e al. © 2019 In e na ional Vi icul u e and Enology Socie y - IVES OENO One 2019, 4, 725-739 726 abou his a ie y in he li e a u e, i s winemaking quali ies and isual simila i ies o Ai én (VIVC 157), he mos wildly g own a ie y o he same egion, i was almos unknown be o e he las wo decades (Hue a e al., 1998; San os e al., 2004). Now, his a ie y is gene ically cha ac e ised and ecognized as au och hone (Ibáñez e al., 2003; Cabello e al., 2012). In his sense, he p esen s udy is he i s mos consis en e o o chemical cha ac e isa ion o an au hen ic Mal a wine. MATERIALS AND METHODS 1. G apes An amoun o 1424 kg o echnologically ipe g apes (22.3 % o o al soluble subs ances) om Vi is ini e a L., c . Mal a was ha es ed a he expe imen al ineya d o he Ins i u o Mad ileño de In es igación y Desa ollo Ru al, Ag a io y Alimen a io (IMIDRA), a m ‘El Encín’, Alcalá de Hena es (Mad id), Spain. G apes we e handpicked and placed in igid s ackable plas ic boxes in amoun s o 18 o 20 kg/box o p e en g ape smashing. A e ha es ing, hey we e s o ed o e nigh in a cooling chambe a 5 ºC o main aining hei quali y un il c ushing. 2. Chemicals and eagen s SO2and analy ical g ade N2 (99.9999 %) we e pu chased om Ca bu os me álicos (Co nellà de Llob ega , Spain), a a ic acid wi h 99.5 % pu i y was om TIDSA (Fuen idueña de Tajo, Spain) and NaOH and NaClO3we e om Scha lab (Ba celona, Spain). Demine alized wa e wi h elec ical conduc i i y o 7 o 8 µS/cm was ob ained by a e e se osmosis uni om Tesacua S.L. (Mad id, Spain). MilliQ-g ade wa e was p oduced in a MilliQ® In eg al 3 pu i ica ion sys em (Me ck Millipo e, Bille ica, USA). Fe michamp ade ma c Saccha omyces ce e isae yeas s we e acqui ed om Oenob ands (Mon pellie , F ance). 3. Winemaking The 1424 kg o g apes we e c ushed and des emmed in a model NDC18 c ushe (Della To ola, T e iso, I aly). The ob ained g ape mash was squeezed in a pilo pneuma ic p ess (Eno Mundi, Za agoza, Spain) o 3 h and SO2was added o i o ob ain 80 mg/L o o al SO2. An amoun o 250 kg o g ape pomace and 1050 L o g ape mus we e ob ained. The g ape mus (wi h a densi y (ρ20ºC) o 1.095 g/mL, o al acidi y o 4.21 g o a a ic acid/L and pH o 3.84) was u he e ige a ed o 4 ºC and kep a his empe a u e o o e nigh o se ling cou se solids. Abou 750 L o he clea pa o he mus we e ans e ed o a 1000 L s ainless s eel essel and 1.5 g/L o a a ic acid we e added o hem o inc ease acidi y. Nex , 150 g o Saccha omyces ce e isae yeas s (Fe michamp om Gis - b ocades, F ance) we e ac i a ed in 1 L o dilu ed wi h wa e mus and inocula ed in he mus o ini ia e e men a ion. Fe men a ion was ca ied ou a 16-18 ºC un il he o al educing suga con en dec eased o 1.6 g/L (i.e. 29 days). Then he clea pa o he wine was ans e ed ( i s acking) o a a iable olume ank wi h loa ing lid and was le o se le o ano he 26 days. Subsequen ly, he clea pa o he wine was ans e ed o ano he essel (second acking) and was le o se le a 4 ºC. Two addi ional ackings we e done a he 65- h and he 80- h days o winemaking and a e ha , he cla i ied wine was s o ed a 12 o 16 ºC o 5 mon hs. 4. Tangen ial- low memb ane il a ion Wine cla i ica ion was ca ied ou by a low p essu e labo a o y TFMF uni , p o ided wi h a 100 L eed ank, a iable low pe is al ic pump (Mas e lex I/P, model 7591-07, Cole-Pa me Ins umen Co., Ve non Hills, Illinois, USA), inle p essu e gauge (0 o 5 ba ), 1” ex e nal diame e single ca idge o hollow ib e ul a il a ion memb anes (RomiconTM, Koch Memb ane Sys ems, Wilming on, MA, USA), ou le p essu e gauge (0 o 5 ba ), needle p essu e con ol al e and a 100 L ecipien essel. Memb anes we e om hyd ophilic polysul one, 500 kDa nominal molecula mass cu o , 0.09 m2o o al il a ion su ace, 1.09 mm inne ib e diame e and 0.45 m o ib e leng h. Comple ely new memb anes we e used in his ea men . Be o e s a ing il a ion, he memb ane ca idge was condi ioned by consecu i e washings wi h demine alised wa e a 20, 50 and 20 ºC. An amoun o 100 L o he cold se led Mal a wine (CSW) was submi ed o ine cla i ica ion by he al eady desc ibed TFMF uni . Fil a ion was ca ied ou by he ba ch concen a ion mode, ecycling he e en a e in he eed ank and collec ing he il a e in o sepa a e essel. The eed low (Q ) was se a 660 L/h (linea eloci y © 2019 In e na ional Vi icul u e and Enology Socie y - IVESOENO One 2019, 4, 725-739 727 o 2.9 m/s), ansmemb ane p essu e (PTM) o 0.7 ba and empe a u e o 14 o 20 ºC (no ex e nal e ige a ion). Fil a ion was s opped when 55 L o cla i ied wine we e p ocessed, ha co esponds o 17 h o il a ion. 5. Wine sampling The cla i ied (memb ane il e ed) wine (MFW) and 10 L o he un il e ed CSW we e bo led wo days a e il a ion and s o ed a he IMIDRA’s wine cella a 11-15 ºC un il hei ins umen al and senso ial analyses. All analyses we e comple ed in he nex h ee mon hs. 6. Memb ane chemical cleaning A e il a ion, memb anes we e washed subsequen ly wi h wa e a 20 ºC o 30 min, and a 50 ºC o 30 min. Regene a ion was ca ied ou wi h NaOH solu ion (pH 11) a 50 ºC o 1 h, NaOH solu ion (pH 11) + 0.1 % sodium hypochlo i e (NaClO) a 50 ºC o 30 min. NaOH was emo ed buy displacemen wi h wa e a 20 ºC un il pH became neu al. 7. Memb ane hyd aulic pe meabili y de e mina ion Memb ane hyd aulic pe meabili y (Lp) was de e mined be o e (L0p) and du ing il a ion (LFp), as well as a e chemical cleaning (LCp) by plo ing he co esponding wa e (wine) lux alues, measu ed a Q o 660 L/h and 20 ºC, e sus he applied TMP, acco ding o Cassano e al. (2011). 8. Physicochemical analysis Hyd ogen ion ac i i y (pH) was measu ed di ec ly by a FE20/EL20 pH-me e (Me le Toledo, Spain). To al soluble subs ances (TSS) we e de e mined by di ec measu emen o wine samples by hand-held e ac ome e A ago (model HSR500, Japan) in he in e al o 0 o 32 % (°B ix). Tu bidi y (haze) was de e mined by a model D-112 u bidime e in he in e al o 0 o 800 NTU (Dinko Ins umen s, Ba celona, Spain). Ti a able acidi y was de e mined acco ding o he me hod o he In e na ional O ganisa ion o Vine and Wine (OIV) (OIV, 2015). To al SO2was de e mined iodome ically as a sum o he ee and combined SO2, acco ding o OIV (OIV, 2009a). To al educing suga s we e de e mined acco ding o he me hod o OIV (OIVb). Alcohol concen a ion was measu ed by ae ome y, acco ding o he me hod o OIV (OIV, 2009c). To al phenρol index (TPI) o he cla i ied wine was de e mined di ec ly by measu emen o he abso p ion o dilu ed (1/100) wi h wa e samples a 280 nm. TPI o non- cla i ied wine samples we e measu ed a e il a ion o he dilu ed (1/100) samples h ough a PVDF memb ane il e wi h a 0.45 µm po e size. Oc anol-wa e pa i ion coe icien (log P) was calcula ed by he so wa e MOE (Chemical Compu ing G oup ULC). To al p o ein con en was assessed by B ad o d’s me hod using Coomassie b illian blue eagen as desc ibed in Salaza e al. (2007) and was exp essed as mg/L o bo ine se um albumin (BSA) (Bio-Rad Labo a o ies Inc, Richmond, CA, USA). P o ein s abili y was ca ied ou by hea es o bo h MFW and CSW wines, acco ding o Li a e al., (2015). In b ie , 10 mL o wine samples we e il e ed h ough a egene a ed cellulose memb ane il e wi h a po e size o 0.45 μm (Wha man, England) and hei u bidi ies we e measu ed by u bidime y. Samples we e hen hea ed a 80 °C o 2 h in a wa e ba h. A e incuba ion, hey we e cooled unde ap wa e , placed in ice and le in e ige a o o o e nigh a 4 °C. Finally, samples we e homogenized by gen le shaking and hei u bidi y was measu ed again. Wines we e conside ed o be p o ein uns able when he di e ence be ween haze be o e and a e hea ing (ne haze) was g ea e han 2 NTU. A oma compounds we e analysed by Pu ge&T ap-GC-MS ollowing he me hodology de eloped by Azna and A oyo (2007). The ap (Voca bTM 3000) con ained di e en adso ben s. Fi s ly, he samples we e dilu ed wi h milliQ- g ade wa e 1/20 ( / ) o a oid sa u a ion o he ap by e hanol and possible p oblems due o oaming. An aliquo o 5 mL was pu ged wi h helium a low a e o 40 mL/min du ing 20 min and deso bed a 250 ºC du ing 10 min. Twen y one ola ile compounds we e analysed: 10 es e s, 7 alcohols, 2 acids, 1 aldehyde and 1 lac one. Iden i ica ion o he compounds was pe o med by compa ison o hei LRI (linea e en ion index) alue and mass spec um wi h hose o pu e e e ence compounds. Quan i ica ion was ca ied ou by ex e nal calib a ion cu es, o each o hese compounds. Odou h eshold (OT) alues o he iden i ied a oma compounds we e aken om Gu h, 1997, López e al., 2002; Azna e al., 2003; Cha ez e al., 2007 and Escude o e al., 2007. Odou ac i i y alues (OAV) we e quan i ied, di iding Ma in P odano e al. © 2019 In e na ional Vi icul u e and Enology Socie y - IVES OENO One 2019, 4, 725-739 728 he concen a ion o each a oma compound by i s OT alue. Indi idual phenolic compounds we e de e mined by HPLC, acco ding o Fe nández de Simón e al., 2003. Losses o each measu ed pa ame e be o e and a e TFMF we e calcula ed acco ding o he nex equa ion: Loss (%) = 100 (CCSW-CMCW)/CCSW, whe e CCSW is he concen a ion o he measu ed pa ame e in he cold se led wine be o e TFMF and CMCW is he concen a ion o he same pa ame e in he il a e o he memb ane cla i ied wine (MCW). Resul s we e epo ed as mean alue ± s anda d de ia ion (SD) o iplica e analysis o each sample. 9. Desc ip i e senso y analysis Desc ip i e senso y analysis was ca ied ou by a panel wi h en ained judges om IMIDRA. This panel has been o med since 2003 by wen y i e as e s a e o y aining sessions (se en mon hs). Selec ion and o ma ion o as e s was ca ied ou acco ding o he in e na ional s anda d ISO 8586:2012. Se en een la ou desc ip o s we e gene a ed using 38 la ou compounds as desc ibed by A oyo e al. (2009) and Balboa-Lagune o e al. (2011). All o hem we e p esen ed o he panellis s a h eshold concen a ion, as well as wi h inc easing concen a ions in a con ol syn he ic wine. Then, as e s we e ained wi h he same compounds added o eal wines om Ai en g apes a di e en concen a ions. Today, he IMIDRA’s senso y panel is o med by nine pe manen and i e a iable as e s. Tas e sessions a e ca ied ou usually once o wice pe week. Senso y e alua ions we e ealized unde ISO s anda ds ela ed o me hodology (ISO 3972:2011), senso y analysis ocabula y (ISO 5492:2008) and as ing oom (ISO 8589:2007). Amoun s o 20 o 30 mL o wine we e se ed in coded s anda d wine as ing glasses a empe a u e o a ound 18 ºC. Judges exp essed hei e alua ion o each ol a i e ( ui y, banana, alcoholic, o - la ou , oxidised a oma, mic obial a oma and o e all a oma in ensi y and quali y) and as e (alcoholic cha ac e , acidi y, ui iness, ege al/g ass, bi e , body, sal y and global as e quali y) desc ip o wi hin a scale om 1 (low in ensi y) o 10 (high in ensi y). Analysis was ca ied ou a duplica e. 10. S a is ical analysis All analy ical da a we e subjec ed o analysis o a iance ANOVA (SPSS, Chicago, Illinois, USA), using he Duncan’s es o he es ima ion o signi ican di e ences be ween samples wi h p obabili y o 99.5 %. RESULTS AND DISCUSSION 1. Winemaking In gene al, he main physicochemical pa ame e s o he s udied Mal a wine accomplish he es ablished o wines om denomina ion o o igin ‘Vinos de Mad id’ speci ic cha ac e is ics (Hue a e al., 1998; Gil e al., 2006; Co de o- Bueso e al., 2013; Co de o-Bueso e al., 2016): d y wine wi h mode a e acidi y, ela i ely low pH and phenolic con en and mode a e o high ui y la ou (Table 1). Howe e , wha de e mines he singula i y o his g ape a ie y om echnological poin o iew was i s high suscep ibili y o oxida ion. This phenomenon can be pe cei ed easily, as i has a di ec e ec on wine colou and a oma: colou changes om yellow/g eenish o g een/g ey and he cha ac e is ic ui y odou dec eases, whe eas some unpleasan oxida i e la ou s a ise. Once s a ed oxida ion, i is di icul o s op i . Tha is why much ca e should be aken o a oid i . Gene ally, se ling and cla i ica ion should be ca ied ou in he sho es possible ime, a as possible as low empe a u es, a oiding manipula ions causing ae a ion and concen a- ions o ee SO2lowe han 30 mg/L. 2. Tangen ial- low memb ane cla i ica ion 2.1. E ec on he main p ocess pa ame e s The kine ics o he il a ion lux du ing he memb ane cla i ica ion p ocess is shown in Figu e 1. Fil a ion s a ed a lux o 52 L/hm2and inished in 17 h a 48 L/hm2. This is a ela i ely low il a ion a e, which can be a ibu ed mos ly o he low ope a ing PTM applied in his ea men . Li e a u e da a (Se ano e al., 1992) show luxes o 57 o 170 L/hm2(a PTM o 0.7 o 1.3 ba ) du ing cla i ica ion o whi e wines and e e o conside ably highe ini ial lux declines o newly used memb anes wi h losses o mo e © 2019 In e na ional Vi icul u e and Enology Socie y - IVESOENO One 2019, 4, 725-739 729 han 30 L/hm2in 1 h (El Rayess e al., 2011). The obse ed e y small ini ial lux decline o he il e ed he e wine can be a ibu ed mos ly o he low load o suspended solids, esul ing a e he exhaus i e cold se ling ca ied ou be o e ine il a ion ( he cold se led wine u bidi y was 24 NTU, Table 1). Figu e 2 shows ha he hyd aulic pe meabili y o he s udied memb ane dec eased down o 139 L/hm2ba du ing he il a ion p ocess, wha means mo e han 75 % loss o pe meabili y. A e chemical cleaning, hyd aulic pe meabili y g ew up o 516 L/hm2ba , wha co esponds o a conside able loss o 20 % o he ini ial wa e lux. The chemical egene a ion p ocedu e was epea ed again, bu i did no imp o e he hyd aulic pe meabili y o he memb ane, which means ha his loss was i e e sible. 3. Quali y o cla i ica ion Quali y o cla i ica ion should be unde s ood as he ex en o emo al o he solid phase om a suspension and is ela ed di ec ly o he u bidi y o he il a e. Good cla i ica ion o whi e g ape mus s and wines is conside ed when hei u bidi y’s a e lowe han 1 NTU (Ribé eau- Gayon e al., 2006a). In his sense, he used he e 500 kDa polysul one memb ane p oduced an excellen cla i ica ion o he ea ed wine, gi ing a u bidi y o 0.11 NTU (Table 1). Ne e heless, quali y c i e ia should conside also a maximum e en ion o he senso y p ope ies and he genuine chemical composi ion o he cla i ied wine. In his con ex and wi h e e ence o he basic physicochemical pa ame e s o he wine, he 500 kDa memb ane did no p oduce any signi ican e ec on mos o he measu ed pa ame e s (Table 1). Only mino diminishing (up o 5.5 %) o he o al wine acidi y and o al phenol index (TPI) we e ound, due mos p obably o he loss o some ola ile acids, such as ace ic and hexanoic acids (Table 2) and some hyd oxycinnamic acids ( ans-Fe a ic and Ca eic), (Table 3), possibly associa ed o he 10.3% loss o he wine p o ein (colloidal) ac ion. The ea men p oduced also a signi ican d op o he hea induced u bidi y (ΔNTU) ( om 2.2 o 0.88 NTU), which is indica i e o some haze s abiliza ion e ec (ΔNTU<2). Ne e heless, his inding has no big ele ance o he wine p o ein s abili y, as he un il e ed wine (CSW) had a ΔNTU alue e y close o he es ablished limi (2.2). Indeed, he accumula ed expe ience wi h Ma in P odano e al. © 2019 In e na ional Vi icul u e and Enology Socie y - IVES OENO One 2019, 4, 725-739 730 FIGURE 1. Pe mea e lux kine ics o Mal a wine du ing cla i ica ion by he 500 kDa hollow ib e memb ane (Q 660 L/h, PTM 0.7 ba , empe a u e 14 °C). TABLE 1. Main physicochemical pa ame e s o he s udied cold se led (CSW) and memb ane il e ed (MFW) wines. Compound CSW MFW pH 3.34 ± 0.02a 3.31 ± 0.02a To al acidi y (g/L) 5.6 ± 0.2a5.2 ± 0.1b Reducing suga s (g/L) 1.6 ± 0.4a1.6 ± 0.2a TPI 5.74 ± 0.01a5.47 ± 0.02b E hanol (%, / ) 12.84 ± 0.03a12.81 ± 0.04a Glyce ol (g/L) 7.85 ± 0.02a7.86 ± 0.04a F ee SO2 (mg/L) 45 ± 3a42 ± 2a To al p o eins (mg/L o BSA) 9.56 ± 0.15a 8.58 ± 0.12b Tu bidi y (NTU) 24.0 ± 0.3a 0.11 ± 0.07b Ne u bididy a e hea es (!NTU) 2.20 ± 0.08a 0.88 ± 0.02b bDi e en le e s wi hin he same ow indica e signi ican di e ences (p ≤ 0.05). Mal a winemaking a he IMIDRA’s wine cella shows ha hese wines ha e high p o ein haze s abili y and usually do no need ben oni e cla i ica ion (no published da a). Mos ele an was he e ec o his ea men on he senso y p ope ies o he s udied wines. Figu e 3 shows he incidence o he TFMF p ocess on he main ol a i e pa ame e s and Figu e 4, on he main as e pa ame e s. Visual e alua ion o he un il e ed Mal a wine showed ha i was eiled o clea wi h mode a ely in ense yellow/g eenish colou (da a no shown). The ol ac i e phase e ealed a high ui y a oma wi h a clea p edominance o banana, and a li le bi lowe in ensi ies o pineapple, apples and pea s. Alcohol odou also was mode a ely app eciable. Nega i e odou cha ac e is ics, such as oxida ion, chemical and mic obial o - la ou s we e ound a e y low le els. The as e phase showed a p edominan ui y la ou wi h clea exp ession o banana a oma on he backg ound o pineapple, apple and pea nuances. Alcohol as e was mode a e and acidi y, balanced. P obably, he mos impo an pa icula i y o his wine was he low in ensi y sal y as e, which appea ed a a la e s age o he as ing, oge he wi h low in ensi y bi e ness. These ine sal y and bi e as es a e e y speci ic o his a ie y and can be used, oge he wi h he high banana a oma, o i s di e en ia ion. Fou o he en as e s de ec ed also he p esence o ace ic acid a oma in he wine. On he o he hand, he TFMF p ocess p oduced a no iceable dec ease o he global a oma quali y and in ensi y (11.4 o 14.5 %, espec i ely), which was exp essed mainly wi h loss o ui iness (15.3 %) and especially, wi h he main ui y componen s: banana (13.2 %), apple and pineapple a oma (da a no shown in his igu e). Howe e , he p ocess p oduced also some posi i e e ec s, educing a small ex en s some o he nega i e ol a i e pa ame e s, mainly hose ela ed o oxida ion (Figu e 3), ege able/ he baceous (Figu e 4) and ace ic acid odou s (Table 2), bu hey we e a e y low de ec ion le els. Wi h espec o he es o he s udied as e pa ame e s, he il a ion p ocess dec eased he global quali y and ui iness o he wine, bu his e ec was no so clea ly de ined by judges, as hose co esponding o he a oma. In gene al, he 500 kDa polysul one memb ane p oduced an © 2019 In e na ional Vi icul u e and Enology Socie y - IVESOENO One 2019, 4, 725-739 731 FIGURE 2. Hyd aulic pe meabili y o he 500 kDa hollow ib e memb ane be o e (L0P) and du ing il a ion (LFP) and a e chemical cleaning (LCP) (Q 660 L/h, empe a u e 20 ºC). FIGURE 3. Main ol a i e pa ame e s o he s udied cold se led (CSW) and memb ane il e ed (MFW) wines. 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Main as e pa ame e s o he s udied cold se led (CSW) and memb ane il e ed (MFW) wines. 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A oma composi ion o Mal a wine be o e (cold se led wine, CSW) and a e memb ane il a ion (memb ane cla i ied wine, MCW), assessed by CG-MS, a oma compound loss (ACloss) a e il a ion, odou h eshold (OT), odou ac i i y alues o CSW and MCW (OAVCSW and OAVMCW), OAV loss (OAVloss) a e il a ion, oc anol-wa e pa i ion coe icien o he s udied molecules (logP (o/w)) and hei molecula masses (MM). aThe same supe sc ip in he same ow means ha he e is no signi ican di e ence be ween alues (p ≤ 0.05). app eciable educ ion o he ui y la ou o he s udied Mal a wine. 4. Incidence on a oma composi ion A oma compounds ha e key con ibu ion o he senso y p ope ies o wines. Twen y one ola ile compounds we e iden i ied and quan i ied in bo h, he cold se led (CSW) and memb ane cla i ied (MCW) Mal a wines by GC-MS analysis (Table 2). Among hem, mos di e se and impo an o he a oma exp ession o he s udied wines was he g oup o he a y acid and alcohol es e s wi h a o al amoun o 25.3 mg/L. The calcula ed odou ac i i y alues (OAV) showed ha Isoamyl ace a e and E hyl oc anoa e had he main impo ance o he global a oma, con ibu ing wi h swee , ui y, banana and pea a omas. These esul s ag ee wi h he a oma ic no es ound du ing he senso y analysis. The E hyl hexanoa e, Hexyl ace a e and E hyl bu y a e had an impo an addi ional con ibu ion o he main a oma bouque wi h esh apple, pineapple, s awbe y and he baceous conno a ions. The e hylphenyl ace a e had a ela i ely low concen a ion, bu enough o gi e some lo al no es o he whole a oma. An impo an numbe o highe alcohols we e also de e mined. They we e ound a highe amoun s (250 mg/L), bu had lowe con ibu ion o he global wine a oma because o hei lowe OAVs. Among hem, only he Isoamyl alcohol and 2-Phenyle hanol had ol a i e signi icance, gi ing usel and lo al conno a ions, espec i ely. O he quan i a i ely impo an componen s, bu wi h lowe odou signi icance we e he Hexanoic and Ace ic acids. The i s one is known o impa unwan ed a y a oma o wines. I s occu ence gene ally is ela ed o he pa ial hyd olysis o E hyl hexanoa e and a highe alues is indica i e o he ini ia ion o p ocesses o oxida ion. The low alues de e mined he e o Hexanoic acid alida e he good s a e o he wine and mean ha i has no con ibu ion o he wine a oma. Ace ic acid was majo ola ile compound wi h an amoun o 443 mg/L. E en i p esence is usually associa ed wi h ace ic acid bac e ia ac i i y, amoun s lowe han 500 mg/L has s ill pleasan odou con ibu ion o he wine a oma. Expe judges e use wines wi h con en s o ace ic acid highe han 600 mg/L (Ede , 2006). Wi h espec o he memb ane cla i ied wine (MCW), i can be seen ha he TFMF p oduced an impo an loss o mos o he de e mined a oma compounds. Mos a ec ed was he g oup o a y acid and alcohol es e s, which ha e he highes con ibu ion o he global wine a oma. Among hem E hyl oc anoa e, E hylphenyl ace a e, Hexyl ace a e, E hyl decanoa e, E hyl hexanoa e, Isoamyl ace a e, Die hyl succina e and E hyl iso ale a e dec eased wi hin he in e al o 43 o 16 %. The a oma ic alcohol 2-Phenyle hanol and he e pene Linalool should be included also in o his g oup because o hei conside able loss o 32 and 18%, espec i ely and because o hei posi i e aid o wine a oma. The loss o hese compounds can be di ec ly ela ed o he dec ease o he pe cep ion o ui iness and a he banana, apple, pea and pineapple conno a ion o he ea ed wine and con i med quan i a i ely he esul s al eady ob ained om he senso y analysis. Con a y o his gene al e ec , he loss o 13 and 19 %, espec i ely, o ace ic and hexanoic acids could be conside ed i no posi i e, a leas , no nega i e, as bo h compounds con ibu e o he wine unwan ed odou s. Ne e heless, i should be men ioned also, ha a conside able dec ease o his acids could be impo an o he global a oma equilib ium, because hey ac also, as a coun e pa in he hyd olysis o he co esponding o hem es e s (Be and, 1981; Edwa ds e al.. 1990). Howe e , i can also be seen ha he il a ion p ocess did no a ec mos o he highe alipha ic alcohols, inc easing hus hei con ibu ion wi h soap, a y, ancid and usel o e he weakened ui y a omas o he memb ane cla i ied wine. I is in e es ing o no e ha hese compounds ha e he smalles molecula masses (up o 102 Da, Table 2), which explain qui e easonably he ob ained esul s. 5. Incidence on phenol composi ion Phenolic compounds ha e impo an con ibu ion o he senso y (acid and bi e as e), mou h eel (as ingency) and unc ional ( edox) p ope ies o wines. Fou een phenolic compounds we e iden i ied and quan i ied in he s udied he e Mal a wines by e e sed phase HPLC analysis (Table 3). As i can be seen om Table 3, he o al amoun o indi idual phenolic compounds in he s udied Mal a wine was ela i ely low: 46.5 mg/L (20.7 mg/L (Table 3) + 25.8 mg/L o 2-Phenyl e hanol and E hylphenyl ace a e, Table 2). This is mos p obably due o he p ope a ie y and/o o he winemaking p ocess ca ied ou in his ial, in which only ee unning g ape juice was used o e men a ion. Mos di e s was he © 2019 In e na ional Vi icul u e and Enology Socie y - IVESOENO One 2019, 4, 725-739 733