Employment of different processes for the production of strawberry vinegars: Effects on antioxidant activity, total phenols and monomeric anthocyanins

Depósi o de in es igación de la Uni e sidad de Se illa h ps://idus.us.es/ “This is an Accep ed Manusc ip o an a icle published by Else ie in LWT- FOOD SCIENCE AND TECHNOLOGY on 2013, a ailable a : h ps://doi.o g/10.1016/j.lw .2012.04.021.” Employmen o di e en p ocesses o he p oduc ion o s awbe y inega s: 1 E ec s on an ioxidan ac i i y, o al phenols and monome ic an hocyanins 2 C. Ubeda1, R. Callejón1, C. Hidalgo2, M.J. To ija2, A.M. T oncoso1, M.L. Mo ales1* 3 1Á ea de Nu ición y B oma ología. Facul ad de Fa macia. Uni e sidad de Se illa. 4 C/ P. Ga cía González nº2, E- 41012. Se illa. Spain. 5 2Depa amen o de Bioquímica y Bio ecnología. Facul ad de Enología. Uni e si a 6 Ro i a i Vi gili. C/ Ma cel·lí Domingo s/n. E- 43007. Ta agona. Spain. 7 *Co esponding au ho : [email p o ec ed]; Tel.: 34-954-556760; Fax.: 34-954-233765 8 Abs ac 9 The use o s awbe y su pluses o he p oduc ion o added alue p oduc s seems o be 10 a good solu ion choice o a oid he was e o his ui . We p oduced s awbe y inega s 11 h ough double e men a ion (alcoholic and ace ous) om h ee di e en ha es s o 12 F aga ia x ananassa a . Cama osa. The objec i e was o s udy he e olu ion o 13 an ioxidan ac i i y, o al phenols and monome ic an hocyanins du ing he inega 14 p oduc ion p ocess. These pa ame e s inc eased when sulphu dioxide and pec oly ic 15 enzymes we e added o subs a es. Inocula ion wi h he Saccha omyces ce e isiae s ain 16 RP1 p oduced wines wi h hal he an hocyanins wi h espec o he spon aneous 17 e men a ions. The use o wood ba els, pa icula ly che y wood ba els, had a posi i e 18 e ec on all he pa ame e s de e mined. All measu ed pa ame e s dec eased du ing he 19 double e men a ion p ocess. In gene al, he ace i ica ion s age led o a high loss o 20 an ioxidan compounds. Mo eo e , he p oduc ion o hese inega s a a semi-pilo scale 21 yielded inal commodi ies wi h he bes alues o an ioxidan ac i i y, o al phenols and 22 monome ic an hocyanins compa ing wi h he inega s ob ained in 2008 and 2009 23 ha es . 24 Keywo ds: an ioxidan ac i i y; monome ic an hocyanins; s awbe y; inega ; wine. 25 2 1. In oduc ion 26 S awbe ies a e a widely esea ched ui o hei nu i ional and heal h bene i s as 27 well as hei o ganolep ic p ope ies. This ui is ich in i amins, mine als, ib e and 28 phy ochemicals. In addi ion, s awbe ies con ain po en ially bioac i e compounds and 29 a e a g ea sou ce o phenolic compounds such as la onoids and phenolic acids (Aaby, 30 K., Sk ede, G., & W ols ad, R. E, 2005; Mää ä-Riihinen, K. R., Kamal-Eldin, A., & 31 Tö önen, A. R, 2004; See am, N. P., Lee, R., Scheulle , H. S., & Hebe , D, 2006). All 32 o hese phenolic compounds ha e been shown o p e en oxida i e p ocesses, 33 pa icula ly hose caused by eac i e oxygen species (ROS) (Aaby, K., Ekebe g, D., & 34 Sk ede, G, 2007; Ce ezo, A. B., Cue as, E., Win e hal e , P., Ga cia-Pa illa, M. C., & 35 T oncoso, A. M, 2010a). These compounds make s awbe ies a highly an ioxidan ui 36 (Aaby e al., 2005; Wol e, K. L., Kang, X., He, X., Dong, M., Zhang, Q., & Liu, R. H, 37 2008) wi h po en ial heal h bene i s. Among he nume ous heal hy p ope ies desc ibed 38 in he li e a u e a e an i-p oli e a i e e ec s on cance cells (Meye s, K. J., Wa kins, C. 39 B., P i s, M. P., & Liu, R. H, 2003; Olsson, M. E., Ande sson, C. S., O edsson, S., 40 Be glund, R. H., & Gus a sson, K, 2006) and he an ioxidan and an i-in lamma o y 41 e ec s ha ha e been shown o educe ca dio ascula disease isk ac o s in se e al 42 p ospec i e coho s udies (Hannum, 2004). 43 Acco ding o he la es da a om he FAO (FAOS a , FAO 2011), Spain is he second- 44 la ges s awbe y p oduce in he wo ld; a la ge po ion o his p oduc ion is ha es ed 45 in Huel a (Andalucía). E e y yea , pa o he c op is disca ded o a ious easons, 46 including size o de o ma ions o he be ies, o o e p oduc ion which leads o 47 su pluses. Because inega is gene ally an inexpensi e p oduc , i s p oduc ion equi es 48 low-cos aw ma e ials, such as sub-s anda d ui and seasonal ag icul u al su pluses 49 (Solie i & Giudici, 2009). In addi ion, he e is a g owing demand o ui inega s, 50 3 which a e sold as a heal h ood (Shau-mei & Chang, 2009). The use o s awbe ies o 51 second quali y, which a e s ill sui able o human consump ion, o p oduc ion heal hy 52 inega s wi h special o ganolep ic nuances may be a good me hod o educe losses due 53 o disca ding he ui . 54 Fo his pu pose, we ha e p oduced s awbe y inega s using second-quali y 55 s awbe ies employing wo-s age e men a ion and assessed di e en condi ions and 56 ea men s. The aim o his wo k was o e alua e he changes in he an ioxidan ac i i y 57 (AA), o al phenols index (TPI) and o al monome ic an hocyanins (TA) du ing he 58 p oduc ion p ocess o s awbe y inega . In addi ion, an adequa e ex ac ion me hod o 59 pe o m hese de e mina ions was designed. 60 2. Ma e ials and me hods 61 2.1. Chemicals 62 The eagen s ace one, me hanol, Folin-Ciocal eu eagen , e hanol, di-po assium 63 hyd ogen phospha e (anhyd ous), sodium di-hyd ogen phospha e 1-hyd a e, po assium 64 chlo ide, sodium ace a e and sodium ca bona e (anhyd ous) we e pu chased om Me ck 65 (Da ms ad , Ge many). Fluo escein sodium and gallic acid we e supplied om Fluka 66 (Mad id, Spain). 6-hyd oxy-2,5,7,8- e ame hylch oman-2-ca boxylic acid (T olox), 67 2,2’-azobis (2-me hylp opionamidine) dihyd ochlo ide (AAPH) and 2,2’-diphenyl-1- 68 pic ylhyd azyl (DPPH) we e pu chased om Sigma-Ald ich (S einheim, Ge many). 69 2.2. Samples 70 Fo he op imisa ion o he ex ac ion p ocess, we used s awbe ies (F aga ia ananassa 71 a . cama osa) acqui ed a he ma ke . The ui was c ushed in ou labo a o y, 72 dis ibu ed in o ambe glass lasks and ozen a -20º C. 73 Fo he p oduc ion o he inega s, we employed h ee di e en ba ches o s awbe ies 74 (F aga ia ananassa a . cama osa) om he Huel a a ea (Spain), co esponding o 75 4 h ee ha es s: 2008, 2009 and 2010. The p oduc ion p ocesses we e pe o med in he 76 labo a o ies o he Dep o Biochemis y and Bio echnology, Facul y o Oenology, Uni 77 Ro i a i Vi gili (Ta agona). In 2008 and 2009, he subs a e employed we e pu ees 78 p epa ed in he labo a o y using a bea e . In 2010, we used a comme cial pu ee p o ided 79 by he Hudisa Company (Huel a). Sulphu dioxide (60 mg/L), suc ose and wo ypes o 80 pec oly ic enzymes (Depec il ex a-ga de FCE® and Depec il cla i ica ion® om Ma in 81 Viala e Oenologie, Epe nay, F ance), bo h a a concen a ion o 15 mg/L, we e added 82 o he pu ee. A e his poin , he p ocedu es we e sligh ly di e en in each ha es . 83 2008 ha es 84 One po ion o he s awbe y pu ee was p essed o s udy he e ec o wo ypes o 85 s a ing subs a es (semi-solid and liquid) (Table 1). Six glass con aine s we e illed 86 wi h 6 L o ui subs a e ( ou pu ees and wo liquids). Hal o he con aine s o each 87 ype o subs a e we e inocula ed wi h he yeas Saccha omyces ce e isiae QA23 a a 88 concen a ion o 2x106 cells/mL, and spon aneous alcoholic e men a ion was allowed 89 o occu in he o he hal . All wines we e spon aneously ace i ied keeping i in he same 90 con aine s. Two inal ea men s we e es ed in inega s: pas eu iza ion o 91 cen i uga ion. The a e age ace ic deg ees in he 2008 s awbe y inega s we e 4.8. 92 2009 ha es 93 Fo he inega p oduc ion in 2009, eigh glass essels we e illed wi h 6 L o 94 s awbe y pu ee each. Hal o hese essels we e inocula ed wi h he yeas s ain 95 Saccha omyces ce e isiae RP1, isola ed du ing he 2008 spon aneous alcoholic 96 e men a ion, and spon aneous alcoholic e men a ion was allowed o occu in he o he 97 hal . All o he wines ob ained om he inocula ed alcoholic e men a ion we e mixed 98 and dispensed in h ee di e en ypes o con aine s: a glass essel and oak o che y 99 wood ba els. Samples we e hen inocula ed wi h a s ain o ace ic acid bac e ia isola ed 100 5 om he 2008 ace i ica ion. Wines om he spon aneous alcoholic e men a ion we e 101 p ocessed in he same way and le o ace i y spon aneously. The inega s ob ained 102 we e pas eu ised. Inocula ed inega s om he 2009 ha es eached an ace ic deg ee o 103 5.5 (glass con aine ), 6.6 (oak ba el) and 6.3 (che y ba el). 104 A po ion o he pu ee om he 2009 s awbe ies was concen a ed by hea ing in a 105 wa e ba h a 80ºC du ing 10 hou s, o es ano he me hod o inc easing he suga 106 con en ; he esul ing p oduc was a cooked mus (Table 1). The suc ose inal 107 concen a ion was 140 g/L. One li e o his subs a e was e men ed by a spon aneous 108 p ocess and one li e was inocula ed wi h he RP1 s ain o yeas . The inocula ed wines 109 (IWs) we e ace i ied wi h he same ace ic acid bac e ia isola ed in 2008, and he 110 spon aneous wines (SWs) we e le o ace i y spon aneously. 111 2010 ha es 112 In his ha es , he pec oly ic enzymes added we e Rohapec ® (12 mg/hL) and he pH 113 was adjus ed o 3.5 wi h 2 g/L CaCO3. In his case, 45 L o pu ee we e e men ed in a 114 s ainless s eel con aine on a semi-pilo scale, a e inocula ion wi h S. ce e isiae RP1. 115 The ace ous e men a ion was pe o med in a che y wood ba el. The inega had an 116 ace ic deg ee o 6.3. 117 All inega s om 2009 and 2010 ha es we e pas eu ized as inal ea men . 118 Fo y-one samples, aken h oughou hese p oduc ion p ocesses, we e analysed. The 119 codes and cha ac e is ics o he samples a e shown in Table 1. In addi ion, i e 120 comme cial inega s we e also analysed o ca y ou compa a i e s udies: Ace o 121 Balsamico, ed wine and whi e wine inega s, apple inega and she y inega . 122 2.3. Sample-ex ac ion p ocedu e 123 The consis ency o he samples (pu ees, wines and inega s) made i necessa y o 124 es ablish an ex ac ion sys em p io o analysis. The me hod employed was based on he 125 6 ex ac ion p ocedu es designed and op imised p e iously by Ubeda, Hidalgo, To ija, 126 Mas, T oncoso & Mo ales (2011a). Twen y g ams o sample we e mixed in a beake 127 wi h 40 ml o ex ac o 10 min while shaking a 800 pm. The sample was hen 128 subjec ed o ul asonica ion ollowed by a cen i uga ion a 4000 pm o 15 min. The 129 supe na an was eco e ed, and he pelle was e-ex ac ed wi h 40 ml o sol en 130 ollowing he same p ocedu e. Bo h ex ac s we e subsequen ly mixed, and he o ganic 131 sol en was emo ed unde acuum. Finally, he ex ac was il e ed, and MilliQ wa e 132 was added o a inal olume o 15 ml. E e y ex ac ion was pe o med in duplica e. We 133 es ed di e en condi ion o ge he maximum alues o AA, TPI and TA as well as 134 economy o sol en used and ime. Thus, he pa ame e s s udied o selec he bes 135 ex ac ion condi ions we e: ype o sol en (ace one, me hanol o e hanol), pe cen age 136 o sol en (80% o 100%) and ul asonic ex ac ion ime (15, 25, 35 o 50 min). 137 2.4. Assays and Me hods 138 2.4.1. ORAC-FL assay 139 The Oxygen Radical Abso bance Capaci y assay (ORAC-FL) was pe o med in a Black 140 96-well mic opla e, ollowing he p ocedu e desc ibed in Ubeda e al. (2011a). This 141 assay was conduc ed in a Mul i-de ec ion pla e eade (Syne gy HT, Ve mon , USA) 142 loca ed a he Cen e o Resea ch, Technology and Inno a ion a he Uni e si y o 143 Se ille (CITIUS). All eac ion assays we e pe o med in iplica e. Resul s we e 144 exp essed as µmol T olox equi alen s (TE)/kg o sample. 145 2.4.2. DPPH adical sca enging assay 146 To de e mine he adical sca enging capaci y, he DPPH assay desc ibed by B and- 147 Williams, Cu elie , & Be se (1995) was used. Fo his es , we used an UV/Vis 148 spec opho ome e U-2800 Digilab coupled o a Pel ie hemos a ic sys em (Hi achi, 149 7 Tokyo, Japan). Resul s we e exp essed as µmol T olox equi alen s (TE)/kg o sample. 150 The assays we e pe o med in iplica e. 151 2.4.3. To al Phenols Index 152 This pa ame e was de e mined in iplica e, using he Folin-Ciocal eu me hod 153 ollowing he p ocedu e desc ibed in Wa e house (2001). Resul s we e exp essed as mg 154 gallic acid/L. 155 2.4.4. To al monome ic an hocyanins 156 The de e mina ion o o al monome ic an hocyanin con en (TA) was measu ed 157 ollowing he pH-di e en ial me hod desc ibed in Gius i & W ols ad (2001). TA was 158 exp essed as pela gonidin-3-glucoside (Plg-3-glu), which is he majo an hocyanin in 159 s awbe y ui wi h a λ is-max a 510 nm (Swain, 1965). Two bu e s we e p epa ed: 160 po assium chlo ide bu e pH=1 (0.025 M), and sodium ace a e bu e pH=4.5 (0.4 M). 161 We measu ed he abso bance a 510 and 700 nm agains a cu e e illed wi h dis illed 162 wa e as a blank. 163 We hen calcula ed he abso bance o he dilu ed sample (A) as ollows: 164 A= (A 510 -A 700) pH 1.0 - (A 510 -A 700) pH 4.5 165 The monome ic an hocyanin concen a ion in he o iginal sample was calcula ed using 166 he ollowing o mula: 167 TA[Plg-3-glu (mg/L)] = (A × MW × DF × 1000) / (ε×1) 168 Whe e 169 A = Sample abso bance 170 MW= Molecula weigh o Plg-3-glu (487.5) 171 DF= Dilu ion ac o 172 ε= Abso p ion coe icien o Plg-3-glu (17330) 173 8 The esul s we e exp essed as mg Plg-3-glu/kg o sample. 174 2.5. S a is ical analysis 175 All s a is ical analysis was pe o med using he S a is ica e sion 7.0 so wa e package 176 (S a so , Tulsa, USA). 177 3. Resul s and discussion 178 3.1. Selec ion o he bes ex ac ion condi ions 179 Se e al ac o s, such as sol en composi ion, ime o ex ac ion, empe a u e, pH, solid- 180 o-liquid a io and pa icle size, may signi ican ly in luence solid-liquid ex ac ions 181 (Azizah, A. H., Ruslawa i, N. M. N., & Tee, T. S, 1999; Pinelo, M., Del Fabb o, P., 182 Manzocco, L., Nunez, M J., & Nicoli, M. C, 2005). In ou case, he pa ame e s ha 183 we e e alua ed o de e mine he bes ex ac ion condi ions we e he ype o sol en , he 184 sol en -wa e a io and ul asonica ion ime. The c i e ia used o selec he ex ac ion 185 pa ame e s we e he maximum alues o an ioxidan ac i i y, o al phenols, 186 an hocyanins and ime and sol en sa ings. 187 The ype o sol en is one o he mos in luen ial a iables in he ex ac ion p ocess. We 188 es ed ace one, e hanol and me hanol. The ex ac ion wi h me hanol ga e he wo s 189 esul s in all he assays. As shown in Figu e 1, ace one yielded he highes alues o 190 DPPH (8327 µmol T olox equi alen s (TE)/kg) and TPI (2090 gallic ac. mg/kg), wi h 191 signi ican di e ences in his las pa ame e . Howe e , we ob ained he bes esul s o 192 he ORAC assay (24329 µmol TE/kg) and o he TA de e mina ion (26.78 mg Plg-3- 193 glu/kg) using e hanol, bu no signi ican di e ences we e ound be ween hese alues 194 and hose wi h ace one (26.30 mg Plg-3-glu/kg). Hen íquez, C., Ca asco-Pozo, C., 195 Gomez, M., B unse , O., & Speisky, H, (2008) epo ed ha he an ioxidan ac i i y o 196 s awbe y ex ac s ob ained wi h ace one/wa e was highe han ha wi h e hanol/wa e 197 and aqueous ex ac s. Taking in o accoun his and o he s udies (Ga cia-Vigue a, C., 198 15 Bosso, A., & Guai a, M. (2008). S udy o some ac o s in ol ed in e hanal p oduc ion 349 du ing alcoholic e men a ion. Eu opean Food Resea ch and Technology, 227, 350 911−917. 351 B and-Williams, W., Cu elie , M. E., & Be se , C. (1995). Use o a ee adical me hod 352 o e alua e an ioxidan ac i i y. LWT-Food Science and Technology, 28, 25-30. 353 Cano-López, M., Pa do-Minguez, F., López-Roca, J. M., & Gómez-Plaza, E. (2006). 354 E ec o mic ooxygena ion on an hocyanin and de i ed pigmen con en and ch oma ic 355 cha ac e is ics o ed wines. Ame ican Jou nal o Enology and Vi icul u e, 57, 325–331. 356 Ce ezo, A. B., Tes aye, W., To ija, M. J., Ma eo, E., Ga cia-Pa illa, M. C., & 357 T oncoso, A. M. (2008). The phenolic composi ion o ed wine inega p oduced in 358 ba els made om di e en woods. Food Chemis y, 109, 606-615. 359 Ce ezo, A. B., Cue as, E., Win e hal e , P., Ga cia-Pa illa, M. C., & T oncoso, A. M. 360 (2010a). Isola ion, iden i ica ion, and an ioxidan ac i i y o an hocyanin compounds in 361 Cama osa s awbe y. Food Chemis y, 123, 574-582. 362 Ce ezo, A. B., Cue as, E., Win e hal e , M., Ga cia-Pa illa, M. C., & T oncoso, A. M. 363 (2010b). An hocyanin composi ion in Cabe ne Sau ignon ed wine inega ob ained by 364 subme ged ace i ica ion. Food Resea ch In e na ional, 43, 1577-1584. 365 Del eil, D., Feuilla , M., Guilloux-Bena ie , M., Sapis, J. C. (2000). Los inos blancos 366 secos. In: C. Flanzy, (ed), Enología: Fundamen os cien í icos y ecnológicos (pp 443- 367 461). Mad id: Vicen e ediciones/Ediciones Mundi-P ensa Publishe s. 368 FAO 2011. URL h p:// aos a . ao.o g/si e/339/de aul .aspx 369 Ga cia-Vigue a, C., Za illa, P., & Tomás-Ba be án, F. A. (1998). The use o ace one as 370 an ex ac ion sol en o an hocyanins om s awbe y ui . Phy ochemical Analysis, 9, 371 274-277. 372 16 Gius i M. M., & W ols ad, R. E. (2001). Cha ac e iza ion and measu emen o 373 an hocyanins by UV–Visible spec oscopy. In R. E. W ols ad, T. E. Ac ee, E. A. 374 Häkkinen, S. H., Heinonen, I. M., Kä enlampi, S. O., Mykkänen, H. M., Ruuskanen, J., 375 & Tö önen, A. R. (1999). Sc eening o selec ed la onoids and phenolic acids in 19 376 be ies. Food Resea ch In e na ional, 32, 345–353. 377 Hannum, S. M. (2004). Po en ial impac o s awbe ies on human heal h: a e iew 378 o he science. C i ical Re iews in Food Science and Nu i ion, 44, 1-17. 379 Hen íquez, C., Ca asco-Pozo, C., Gomez, M., B unse , O., & Speisky, H. (2008). 380 Slow and as - eac ing an ioxidan s om be ies: hei e alua ion h ough he 381 FRAP (Fe ic Reducing An ioxidan Powe ) assay. Ac a Ho icul u ae, 777, 531- 382 536. 383 Klopo ek, Y., O o, K., & Boehm, V. (2005). P ocessing s awbe ies o di e en 384 p oduc s al e s con en s o i amin C, o al phenolics, o al an hocyanins, and 385 an ioxidan capaci y. Jou nal o Ag icul u al and Food Chemis y, 53, 5640-5646. 386 Lee, J., & W ols ad, R. E. (2004). Ex ac ion o an hocyanins and polyphenolics om 387 bluebe y-p ocessing was e. Jou nal o Food Science, 69, 564-573. 388 Mää ä-Riihinen, K. R., Kamal-Eldin, A., & Tö önen, A. R. (2004). Iden i ica ion and 389 quan i ica ion o phenolic compounds in be ies o F aga ia and Rubus species ( amily 390 Rosaceae). Jou nal o Ag icul u al and Food Chemis y, 52, 6178-6187. 391 Meye , A. S. (2002). Enhanced ex ac ion o an ioxidan phenols om wine and 392 juice p ess esidues ia enzyma ic polysaccha ide hyd olysis. F ui P ocessing, 12, 393 29-33. 394 Meye s, K. J., Wa kins, C. B., P i s, M. P., & Liu, R. H. (2003). An ioxidan and 395 an ip oli e a i e ac i i ies o s awbe ies. Jou nal o Ag icul u al and Food 396 Chemis y, 51, 6887-6892. 397 17 Mo a a, A., Gomez-Co do es, M. C., Colomo, B., & Sua ez, J. A. (2005). Cell wall 398 an hocyanin adso p ion by di e en Saccha omyces s ains du ing he e men a ion o 399 Vi is ini e a L. c G aciano g apes. Eu opean Food Resea ch and 400 Technology, 220, 341-346. 401 Olsson, M. E., Ande sson, C. S., O edsson, S., Be glund, R. H., & Gus a sson, K. 402 (2006). An ioxidan le els and inhibi ion o cance cell p oli e a ion in i o by 403 ex ac s om o ganically and con en ionally cul i a ed s awbe ies. Jou nal o 404 Ag icul u al and Food Chemis y, 54, 1248-1255. 405 Pinelo, M., Del Fabb o, P., Manzocco, L., Nunez, M J., & Nicoli, M. C. (2005). 406 Op imiza ion o con inuous phenol ex ac ion om Vi is ini e a byp oduc s. Food 407 Chemis y, 92, 109-117. 408 See am, N. P., Lee, R., Scheulle , H. S., & Hebe , D. (2006). Iden i ica ion o phenolic 409 compounds in s awbe ies by liquid ch oma og aphy elec osp ay ioniza ion mass 410 spec oscopy. Food Chemis y, 97, 1-11. 411 Shau-mei, A., & Chang R. C. (2009). Taiwan ui inega . In L. Solie i, & P. Giudici 412 (Eds.), Vinega s o he Wo ld (pp. 223-242). Milan: Spinge -Ve lag I alia. 413 Solie i, L., & Giudici, P. (2009). Vinega s o he wo ld. In L. Solie i, & P. Giudici 414 (Eds.), Vinega s o he Wo ld (pp. 1-16). Milan: Spinge -Ve lag I alia. 415 Swain, T. (1965) Analy ical me hods o la onoids. In T. W. Goodwin (Ed.), 416 Chemis y and Biochemis y o Plan Pigmen s (pp. 543-544). London: Academic P ess. 417 Ubeda, C., Hidalgo, C., To ija, M. J., Mas, A., T oncoso, A. M., Mo ales, M. L. 418 (2011a). E alua ion o an ioxidan ac i i y and o al phenols index in pe simmon 419 inega s p oduced by di e en p ocesses. LWT-Food Science and Technology, 44, 420 1591-1596. 421 18 Ubeda, C., Callejón, R. M., Hidalgo, C., To ija, M. J., Mas, A., T oncoso, A. M., 422 Mo ales, M. L. (2011b). De e mina ion o majo ola ile compounds du ing he 423 p oduc ion o ui inega s by s a ic headspace gas ch oma og aphy–mass spec ome y 424 me hod. Food Resea ch In e na ional 44, 259–268. 425 Ve beys , L., Oey, I., Van de Plancken, I., Hend ickx, M., & Van Loey, A. (2010). 426 Kine ic s udy on he he mal and p essu e deg ada ion o an hocyanins in s awbe ies. 427 Food Chemis y, 123, 269-274. 428 Wol e, K. L., Kang, X., He, X., Dong, M., Zhang, Q., & Liu, R. H. (2008). Cellula 429 an ioxidan ac i i y o common ui s. Jou nal o Ag icul u al and Food Chemis y, 56, 430 8418-8426. 431 Xu, Q., Tao, W., & Ao, Z. (2007). An ioxidan ac i i y o inega melanoidins. Food 432 Chemis y, 102, 841-849. 433 434 435 436 437 438 439 440 441 442 443 444 445 446 19 447 448 Figu e cap ions 449 Figu e 1. ORAC, DPPH (le axis) and TPI ( igh axis) alues o he di e en 450 ex ac ion sol en s es ed in s awbe ies acqui ed a he ma ke . The ba s in he same 451 assay wi h di e en le e s show signi ican di e ences (p<0.05) (ORAC assay: a, b, c; 452 IPT: A, B, C; DPPH es : α, β, γ). 453 Figu e 2. E ec o sol en pe cen ages. a) ORAC and DPPH alues. b) TPI and 454 TA alues o s awbe ies acqui ed a he ma ke . The ba s in he same assay wi h 455 di e en le e s show signi ican di e ences (p<0.05) (ORAC and TPI assays: a, b; 456 DPPH and TA es s: A, B). 457 Figu e 3. E ec o di e en ul asonica ion imes a) ORAC and DPPH 458 alues. b) TPI and TA alues o s awbe ies acqui ed a he ma ke . The 459 ma ke s in he same assay wi h di e en le e s show signi ican di e ences (p<0.05) 460 (ORAC and TPI assays: a, b, c; DPPH and TA es s: A, B, C). 461 Figu e 4. Compa ison o ORAC, DPPH (le axis) and TPI ( igh axis) alues o 462 s awbe y inega s wi h comme cial a ie ies. Sample codes: F9MCV (mean alue o 463 all inega s om cooked mus ), F9V (mean alue o all inega s om 2009 ha es ) 464 and F8V (mean alue o all inega s om 2008 ha es ). 465 466 467 468 469 470 471 20 Figu e 1. 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486 Figu e 2. 487 488 489 Figu e 3. 490 491 492 493 494 495 496 497 a b a α ß α C B A 0 5000 10000 15000 20000 25000 30000 Ace one Me hanol E hanol µmol TE/kg R 0 500 1000 1500 2000 2500 mg gallic acid/kg b a B A 0 5000 10000 15000 20000 25000 80% 100% µmol TE/Kg 0 1000 2000 3000 4000 5000 6000 7000 8000 9000 µmol TE/Kg a b A B 0 500 1000 1500 2000 2500 80% 100% mg gallic acid/Kg 0 5 10 15 20 25 30 mg Plg-3-glu/Kg a) b) a) b) b b a a C B A A 22000 23000 24000 25000 26000 27000 28000 29000 15 25 35 50 µmol TE/Kg 5000 5500 6000 6500 7000 7500 8000 8500 9000 9500 µmol TE/Kg a b c d A B A A 2000 2100 2200 2300 2400 2500 2600 2700 15 25 35 50 mg gallic acid/Kg 40 45 50 55 60 65 70 mg Plg-3-glu/Kg a) b) 21 Figu e 4. 498 499 500 501 502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 521 522 523 0 5000 10000 15000 20000 25000 30000 35000 40000 45000 Balsamic F9MCV F10VI F9V F8V Apple She y wine Red wine Whi e wine µmol TE/kg 0 500 1000 1500 2000 2500 3000 mg gallic acid/kg 32 Table 1. Samples desc ip ion. Ha es T ea men Pu ee Sample T ea men Sample subs a e Alcoholic e men a ion ( ime) Wine Sample Ace i ica ion ( ime) T ea men o Recipien Vinega sample 2008 C ushed F8P1 SO2 Pec oly ic enzymes Suc ose (50 g/L) F8P2 Inocula ed (4 days) F8WI1- F8WI4 Spon aneous (2 mon hs) Cen i uga ion F8VIC1-F8SVIC2 Pas eu iza ion F8SVIP1-F8SVIP2 Spon aneous (5 days) F8WE1- F8WE4 Cen i uga ion F8SVEC1-F8SVEC2 Pas eu iza ion F8SVEP1-F8SVEP2 - F8P2 P essing F8L Inocula ed (4 days) F8LWI - - - Spon aneous (5 days) F8LWE 2009 C ushed F9P1 SO2 Pec oly ic enzymes Suc ose (75 g/L) F9P2 Inocula ed (5 days) F9WI1- F9WI4 Inocula ed (2 mon hs) glass essel F9SVIG oak ba el F9SVIO che y ba el F9SVIX Spon aneous (8 days) F9WE1- F9WE4 Spon aneous (2 mon hs) glass essel - oak ba el - che y ba el - Hea ing Concen a ed F9MC Inocula ed (7 days) F9MCWI1-F9MCWI2 Inocula ed (5 mon hs) glass essel F9MCVI1-F9MCVI2 Spon aneous (7 days) F9MCWE1-F9MCWE2 Spon aneous (2.5 mon hs) glass essel F9MCVE1-F9MCVE2 2010 C ushed F10P1 SO2 Pec oly ic enzymes Suc ose (65 g/L) CaCO3 F10P2 Inocula ed (4 days) F10WI Inocula ed (1.5 mon hs) che y ba el F10VI 33 Table 2. Changes in 2008 samples on ORAC, DPPH, TPI and TA du ing s awbe y inega p oduc ion (a e age±s anda d de ia ion). 1 Samples ORAC (μmol TE/kg) DPPH (μmol TE/kg) TPI (mg gallic acid /kg) TA (mg plg-3-glu/kg) Subs a es F8P1 21792 ± 221 8327 ± 99 2090 ± 10 26.3 ± 0.8 F8P2 26714 ± 910a 10116 ± 88a 2298 ± 0a 69 ± 0a F8L 20642 ± 111b 5907 ± 516b 1615 ± 33b 43 ± 0b Wines F8LWE 12757 ± 267b,c 2837 ± 59b,c 868 ± 29b,c 12.2 ± 0.2b F8LWI 13497 ± 227b,c 2898 ± 129b,c 858 ± 13b,c 17.9 ± 0.2b,d F8SWE1 25314 ± 650 8200 ± 58b 1907 ± 26 13.1 ± 0.7b F8SWE2 24696 ± 70 7879 ± 70b 1773 ± 32 12.9 ± 0.6b F8SWE3 25458 ± 403 7689 ± 82b 1757 ± 45 12.4 ± 0.7b F8SWI1 27987 ± 1227b 7241 ± 35b,d 1670 ± 9b,d 16 ± 0b,d F8SWI2 25451 ± 429b 8004 ± 35b,d 1584 ± 19b,d 18.0 ± 0.3b,d F8SWI3 23745 ± 15b 6515 ± 67b,d 1548 ± 6b,d 17.3 ± 0.6d Vinega s F8SVE1C 9202 ± 390b 3256 ± 205b 769 ± 13b 0.4 ± 0.0b F8SVE1P 9849 ± 413b 3368 ± 352b 774 ± 23b 0.5 ± 0.1b F8SVE2C 9215 ± 338b 3210 ± 129b 781 ± 0b 1.1 ± 0.2b F8SVE2P 10869 ± 190b 3252 ± 234b 683 ± 10b 0.6 ± 0.0b F8SVI1C 10139 ± 341b,e 3227 ± 117b 751 ± 16b 1.3 ± 0.0b F8SVI1P 11611 ± 89b,e 3388 ± 64b 744 ± 6b 0.9 ± 0.1b F8SVI2C 11054 ± 40b,e 3260 ± 246b 694 ± 16b 0.8 ± 0.1b F8SVI2P 11082 ± 86b,e 3380 ± 76b 712 ± 9b 1 ± 0b 34 Sample codes a e loca ed in Table 1. a Signi ican di e ences (p<0.05) wi h espec o he ini ial ui pu ee (ANOVA). b Signi ican di e ences (p<0.05) wi h espec o he sample om which was p oduced (ANOVA). c Signi ican di e ences (p<0.05) wi h espec o semisolid wines ob ained wi h simila alcoholic p ocess (spon aneous o inocula ed) (ANOVA). d Signi ican di e ences (p<0.05) wi h espec o spon aneous p ocess (ANOVA). e Signi ican di e ences (p<0.05) wi h espec o he inega s ob ained om spon aneous wines (ANOVA). 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17