Thermal extraction and characterization of pectin from semi-solid by-products of the olive oil industry

Resea ch A icle Recei ed: 2 Janua y 2025 Re ised: 28 Ma ch 2025 Published online in Wiley Online Lib a y: (wileyonlinelib a y.com) DOI 10.1002/js a.14316 The mal ex ac ion and cha ac e iza ion o pec in om semi-solid by-p oduc s o he oli e oil indus y A ica Fe nandez-P io ,a,b * Luna Ba e a-Chamo o,a,b El i a Ma quez-Pa adas,a,b Ca ol López-de-Dicas illo,c Ma ia C. Millan-Lina es,dAl a o Villanue a-Lazodand Se gio Mon se a -de la Paza,b Abs ac BACKGROUND: The alo iza ion o ag o-indus ial by-p oduc s is c ucial o p omo ing sus ainabili y and ci cula economy. Oli e mill semi-solid by-p oduc s (OMSbP), also known as alpe ujo, con ain aluable bioac i e compounds, including pec in, which can be ex ac ed and used in ood applica ions. Howe e , he s uc u al p ope ies an an ioxidan po en ial o hese pec- in equi e u he cha ac e iza ion o assess hei po en ial as unc ional ing edien s. RESULTS: In he p esen s udy, pec in was ex ac ed om OMSbP using a ci ic acid-based he mal ex ac ion p ocess. The yield a ied depending on he oli e a ie y and ipeness deg ee, wi h highe ex ac ion e ficiency in mo e ma u e samples (48.2 g kg −1 ). S uc u al analysis by a enua ed o al eflec ance/Fou ie ans o m-in a ed spec oscopy confi med a high homogalac u onan con en (∼500 g kg −1 ) and a deg ee o me hyl es e ifica ion o app oxima ely 60%. The monosaccha ide p ofile indica ed en ichmen in a abinose, hamnose, galac ose and mannose. An ioxidan ac i i y, assessed h ough DPPH (i.e. 2,2-diphenyl-1-pic ylhyd azyl) adical sca enging, showed a s ong co ela ion wi h mannose con en ( =−0.9967). Addi- ionally, pec in-based films demons a ed he mal s abili y compa able o comme cial pec in used in ood packaging. CONCLUSION: This s udy highligh s he po en ial o OMSbP-de i ed pec in as a bio unc ional ing edien wi h an ioxidan ac i - i y and he mal s abili y. The ex ac ion me hod ensu es a sus ainable app oach o u ilizing oli e indus y by-p oduc s. Fu u e esea ch should explo e i s bioa ailabili y and applica ion in edible films and coa ings. © 2025 The Au ho (s). Jou nal o he Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y. Keywo ds: oli e oil semi-solid by-p oduc s; Olea eu opaea; s uc u al cha ac e iza ion; an ioxidan ac i i y; ood packaging INTRODUCTION The inc easing en i onmen al challenges humani y aces oday a e d i ing he de elopmen o mo e sus ainable p oduc ion and consump ion sys ems. Biomass esou ces, as a esul o hei abundance, ep esen a aluable oppo uni y o he c ea ion o high- alue p oduc s. 1 In he oli e oil sec o , he wo-phase ex ac- ion sys em gene a es a semi-solid by-p oduc known as oli e mill semi-solid by-p oduc (OMSbP) o alpe ujo. 2-4 This by-p oduc , p ima ily composed o oli e peel, pulp, seeds and pi agmen s, con ains aluable bioac i e compounds such as polyphenols and ca bohyd a es, including cellulose, hemicellulose and pec- in. 5 Since he 1990s, OMSbP has played a ole in educing he en i onmen al impac o oli e oil ex ac ion by minimizing liquid was e accumula ion. Howe e , i s high mois u e con en p esen s challenges o e ficien managemen . Recen indus ial he mal p e- ea men s ha e enhanced he ex ac ion o bioac i e *Co espondence o: A Fe nandez-P io , Depa men o Medical Biochemis y, Molecula Biology, and Immunology, School o Medicine, Uni e si y o Se ille, A . Sanchez Pizjuan s/n, 41009 Se ille, Spain, E-mail: m e [email p o ec ed] (Fe nandez-P io ) aDepa men o Medical Biochemis y, Molecula Biology and Immunology, School o Medicine, Uni e si y o Se ille, Se ille, Spain bIns i u o de Biomedicina de Se illa, IBiS/Hospi al Uni e si a io Vi gen del Rocio/CSIC/Uni e sidad de Se illa, Se ille, Spain cPackaging G oup, Ins i u o de Ag oquimica y Tecnologia de los Alimen os, Spanish Na ional Resea ch Council (IATA-CSIC), Pa e na, Spain dDepa men o Food & Heal h, Ins i u o de la G asa, Spanish Na ional Resea ch Council (IG-CSIC), Se ille, Spain © 2025 The Au ho (s). Jou nal o he Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y. This is an open access a icle unde he e ms o he C ea i e Commons A ibu ion-NonComme cial-NoDe i s License, which pe mi s use and dis ibu ion in any medium, p o ided he o iginal wo k is p ope ly ci ed, he use is non-comme cial and no modifica ions o adap a ions a e made. 1 compounds, including hyd oxy y osol and a ious ca bohyd a e ac ions. This p og ess ma ks a significan s ep owa d es ablish- ing an OMSbP bio efine y, whe e op imizing ca bohyd a e ex ac- ion and u iliza ion emains a key objec i e. Pec in, a complex he e opolysaccha ide, is composed mainly o homogalac u onan (HG) domains and hamnogalac u onan I (RG-I) egions wi h a galac u onic acid (GalA) backbone. The GalA uni s may be subs i u ed wi h me hyl g oups, as some o he C-6 ca boxyl g oups a e me hyl-es e ified. This es e ifica ion influ- ences he s uc u al cha ac e is ics and physicochemical p ope - ies o pec in, he eby de e mining i s unc ionali y in ood and o he indus ial applica ions. Found in he p ima y cell wall and middle lamella o plan issues, pec in con ibu es signifi- can ly o plan s uc u e and in eg i y. 6 OMSbP-de i ed pec in has exhibi ed no able biological ac i i ies, including an ioxidan , an ip oli e a i e and an i-inflamma o y e ec s, highligh ing i s po en ial as a unc ional ing edien . 7-12 The inc easing demand o na u al ood addi i es is d i ing in e es in sus ainable sou ces o pec in. 13 The p esen s udy in es iga es he ex ac ion and cha ac e iza ion o pec in om OMSbP using a ci ic acid-based he mal me hod, analyzing i s s uc u al p ope ies, an ioxidan capaci y and po en ial applica- ions in ood echnology. MATERIALS AND METHODS Raw ma e ial OMSbP (i.e. alpe ujo), a semi-solid by-p oduc composed o oli e peel, pulp, seeds, and pi agmen s, was ob ained om Almaza a de la Subbe ica S.C.A. (Co doba, Spain). OMSbP samples we e collec ed om wo di e en oli e a ie ies: A bequina and Hoji- blanca du ing he ea ly ha es season o 2022–2023 (designa ed as aeOMSbP and heOMSbP, espec i ely). Addi ionally, samples we e aken om a mix u e o hese a ie ies a wo di e en ip- ening s ages du ing he same ha es season: an ea ly-s age sam- ple (meOMSbP) and a ully ma u e sample (mmOMSbP). Samples we e collec ed di ec ly a he end o he ho izon al cen i uge in he wo-phase ex ac ion sys em and s o ed a −20 °C un il u - he p ocessing. P epa a ion o cell wall ma e ial and pec in ex ac ion OMSbP samples we e p ocessed ollowing he Rena d me hod wi h modifica ions o ob ain ex ac ed pec in. 14 B iefly, 1 L o 800 g kg −1 e hanol was added o 1 kg o esh OMSbP, and he mix u e was incuba ed o e nigh unde con inuous shaking a oom empe a u e. A e cen i uga ion a 15000 ×g o 20 min (RC5C; So all, Mechelen, Belgium), he supe na an was dis- ca ded, and he emaining alcohol-insoluble esidue (AIR) was d ied a 40 °C o 24 h. Pec in ex ac ion was pe o med by sus- pending 30 g o AIR in 300 mL o 30 g kg −1 ci ic acid solu ion, adjus ing he pH o 3.0, and s i ing a 95 °C o 95 min. 15 The mix- u e was hen cen i uged a 3700 ×ga 4 °C o 15 min, and he supe na an was ea ed wi h e hanol (2:1 / ) and s o ed a 4 °C o 16 h o p ecipi a e pec in. A e an addi ional cen i uga ion s ep a 3700 ×g o 30 min a 4 °C, he p ecipi a e was washed h ee imes wi h anhyd ous e hanol and eeze-d ied. Chemical cha ac e iza ion o ex ac ed pec in Pec in yield was calcula ed as he a io o he d y ex ac ed pec in o he o iginal AIR weigh , exp essed as g kg −1 d y OMSbP. The neu al suga con en (NSC) was de e mined using he An h one me hod, as desc ibed by Dishe. 16 B iefly, 100 μLo sample ( iplica e) was mixed wi h 200 μL o 0.2% (w/ ) an h one in concen a ed sul u ic acid, hea ed a 100 °C o 5 min, and mea- su ed a 630 nm in a mic opla e eade (iMa k; Bio-Rad, He cules, CA, USA). Glucose was used as he s anda d (0.02–0.2 mg mL −1 ). U onic acid con en was quan ified using he m-hyd oxydiphenyl me hod, as desc ibed by Blumenk an z and Asboe-Hansen. 17 Samples (200 μL, iplica e) was mixed wi h 1.2 mL o 0.0125 Msodium e abo a e in sul u ic acid, hea ed a 100 °C o 5 min, and eac ed wi h 20 μL o 0.15% m- hyd oxybiphenyl in 0.5% NaOH. Abso bance was measu ed a 520 nm using a galac u onic acid s anda d. Glycosidic composi- ion was analyzed by gas ch oma og aphy (GC, HP6890 Plus; Hew- le Packa d, Palo Al o, CA, USA) a e he de i a iza ion in o aldi ol ace a es, wi h ini ial ifluo oace ic acid hyd olysis using inosi ol as he in e nal s anda d. 18 Quan ifica ion was pe o med using s anda d solu ions o L- hamnose (Rha), L-a abinose (A a), D- galac ose (Gal), D-glucose (Glc), D-mannose (Man), D-Fucose (Fuc) and D-xylose (Xyl). The ch oma og aphic condi ions we e desc ibed by Lama-Muñoz e al. 19 The o al phenolic con en (TPC) was de e mined using he Folin–Ciocal eu spec opho o- me ic me hod wi h sligh modifica ions. 20 B iefly, 20 μL o each sample was mixed wi h 100 μL o Folin–Ciocal eu eagen and 80 μL o a 0.70 Msodium ca bona e solu ion in wa e . The mix u e was s i ed and incuba ed o 10 min o allow colou de elop- men . Abso bance was measu ed a 655 nm. A calib a ion cu e was es ablished using gallic acid as he s anda d. The p o ein con- en o he samples was de e mined using a ni ogen elemen a y analyze (LECO, S Joseph, MI, USA), based on in e nal combus ion a 1000 °C ollowed by he mal conduc i i y de ec ion. The p o- ein con en (g kg −1 o d y ex ac ) was calcula ed using he con- e sion ac o 6.25, in acco dance wi h he ISO me hod. 21 DPPH adical sca enging capaci y Lyophilized pec in samples we e dissol ed in wa e a 10 mg mL −1 and fil e ed h ough a 0.45-μm nylon fil e . The an i- oxidan ac i i y was assessed using he 2,2-diphenyl- 1-pic ylhyd azyl (DPPH) adical sca enging me hod, 22 whe e 5μL o each ex ac was added o 195 μL o 3.8 mg pe 50 mL DPPH solu ion in me hanol, incuba ed o 30 min a oom empe - a u e, and measu ed a 490 nm. The esul s we e exp essed as EC 50 alues (mg mL −1 ), ep esen ing he concen a ion equi ed o educe adical ac i i y by 50%. Deg ee o me hyl es e ifica ion by ATR/FT-IR and 1 H NMR spec oscopy ATR/FT-IR spec oscopy was conduc ed o de e mine he deg ee o me hyl es e ifica ion o he ex ac ed pec in. Th ee eplica es o each sample we e analyzed, and peak in ensi y was used o measu emen . This me hod p o ides a eliable quali a i e and semi-quan i a i e assessmen o pec in es e ifica ion, as epo ed by Muñoz-Almag o e al. 23 The samples we e g ound in an aga e mo a o p ope homogeniza ion be o e analysis. Spec a we e eco ded using a FT/IR-4700 spec opho ome e (Jasco, Tokyo, Japan) ac oss he ange 400–4000 cm −1 . Deg ee o me hyl es e - ifica ion (DM) was calcula ed using he a io o he abso p ion bands a 1730 and 1620 cm −1 :DM=A 1730 /(A 1730 +A 1620 ). 1D 1 H NMR spec oscopy o meOMSbP and mmOMSbP sam- ples was also ca ied ou o de e mine hei s uc u al cha ac e is- ics. Each pec in samples was dissol ed in D 2 O, and he 1 H NMR spec a we e eco ded a 25 °C using an A ance NEO 500 MHz spec ome e (B uke , Bille ica, MA, USA) equipped wi h a Onebay www.soci.o g A Fe nandez-P io e al. wileyonlinelib a y.com/js a © 2025 The Au ho (s). Jou nal o he Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y. J Sci Food Ag ic 2025 2 10970010, 0, Downloaded om h ps://scijou nals.onlinelib a y.wiley.com/doi/10.1002/js a.14316 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [17/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License console ea u ing h ee adio equency channels and a BBFO 5 mm dual esonance Z-g adien p obe. Su ace mo phology analysis by scanning elec on mic oscopy (SEM) Lyophilized pec in samples we e analyzed by SEM o examine su - ace mo phology. The samples we e spu e -coa ed wi h a hin laye o gold–palladium unde acuum using an EM ACE600 ins umen (Leica Mic osys ems, We zla , Ge many) o imp o e conduc i i y. Imaging was pe o med wi h a Zeiss C ossbeam 550 scanning elec on mic oscope (Zeiss, Mad id, Spain) a an accele a ing ol age o 2.00 kV and di e en magnifica ions. 24 The mog a ime ic analysis o pec in films Pec in films om meOMSbP and mmOMSbP samples we e ob ained ia he cas ing echnique, whe e 20 g kg −1 pec in dis- pe sions in dis illed wa e we e pou ed in o Pe i dishes and d ied a 37 °C o e nigh . The esul ing films ( hickness: 45–55 μm) we e analyzed o he mal s abili y using he mog a ime ic analysis (TGA) Q-500 ins umen (TA Ins umen s, New Cas le, DE, USA). App oxima ely 5–8 mg o each sample was placed in alumina capsules and subjec ed o a empe a u e ange o 30–800 °Ca a hea ing a e o 10 °C min −1 unde ni ogen a mosphe e (50 mL min −1 ). The maximum deg ada ion empe a u es (T d ) was de e mined. S a is ical analysis Pea son's co ela ion coe ficien s we e calcula ed o den i y sig- nifican ela ionships be ween pec in yield, chemical compo- nen s, molecula ea u es and an ioxidan ac i i y. Co ela ion hea maps we e gene a ed using he ggplo 2 package (Wickham H, ggplo 2: Elegan G aphics o Da a Analysis. Sp inge -Ve -lag, New Yo k (2016)). ( 3.4.2). h ps://link.sp inge . com/book/10.1007/978-3-319-24277-4. Linea eg ession models we e used o e alua e he p edic i e powe o specific a iables o an ioxidan ac i i y, wi h he coe ficien o de e mina ion (R 2 ) used o assess model fi . All alues a e exp essed as he mean ±SD. The numbe o eplica es used o each measu emen is explici ly s a ed o ensu e anspa ency and ep oducibili y. S a is- ical analyses we e pe o med using P ism, e sion 8.01 (G aphPad So wa e Inc., San Diego, CA, USA). Significance be ween sample g oups was de e mined using a pai ed - es o equal a iances. P<0.05 was conside ed s a is ically significan . RESULTS AND DISCUSSION Chemical cha ac e iza ion o ex ac ed pec in om OMSbP Ex ac ed pec in om OMSbP using ci ic acid a 95 °C om di e - en oli e a ie ies (i.e., aeOMSbP e sus heOMSbP) and ipeness s ages (i.e., meOMSbP e sus mmOMSbP) we e ully cha ac e ized o i s chemical composi ion, as shown in Table 1. F eezing he Table 1. Yield, chemical composi ion and molecula ea u es o OMSbP pec in om wo di e en oli e a ie ies (aeOMSbP and heOMSbP) and wo di e en ipeness s ages (meOMSbP and mmOMSbP) Ex ac ed pec in om oli e mill semi-solid by-p oduc (OMSbP) aeOMSbP heOMSbP P- alue meOMSbP mmOMSbP P- alue Yield (g kg −1 ) a,b 24.80 ±1.90 34.90 ±2.90 0.007 40.00 ±3.10 48.20 ±3.90 0.046 P o eins b 27.40 ±1.90 33.10 ±0.40 0.007 18.50 ±1.50 12.20 ±0.10 0.002 Phenols b 7.80 ±0.10 6.40 ±0.60 0.016 07.20 ±0.20 5.30 ±0.10 <0.001 To al ca bohyd a es b 715.40 ±9.70 570.10 ±7.10 <0.001 570.70 ±20.60 504.90 ±0.90 0.005 Monosaccha ide composi ion c Galac u onic acid (GalA) 62.48 ±0.17 52.20 ±0.07 <0.001 51.63 ±0.29 50.03 ±0.16 0.001 Rhamnose (Rha) 1.59 ±0.10 1.71 ±0.04 0.126 1.95 ±0.24 0.69 ±0.03 0.001 Fucose (Fuc) 0.34 ±0.02 0.65 ±0.07 0.002 0.58 ±0.06 0.54 ±0.18 0.734 A abinose (A a) 7.55 ±0.48 9.05 ±1.41 0.156 10.30 ±0.28 13.58 ±3.04 0.136 Xylose (Xyl) 4.70 ±0.37 4.37 ±1.03 0.629 4.75 ±0.83 3.68 ±0.58 0.141 Mannose (Man) 2.81 ±0.14 3.89 ±0.34 0.007 4.24 ±0.24 4.79 ±1.49 0.562 Galac ose (Gal) 10.71 ±0.96 9.81 ±1.62 0.454 12.97 ±2.10 13.04 ±1.25 0.963 Glucose (Glc) 9.82 ±0.67 10.62 ±1.67 0.484 13.58 ±1.07 13.65 ±1.20 0.943 Mola a io d Homogalac u onan (HG) 60.89 ±0.10 50.49 ±0.04 <0.001 49.68 ±0.24 49.34 ±0.03 0.072 Rhamnogalac u onan I (RGI) 21.45 ±1.39 22.27 ±3.15 0.701 27.18 ±2.84 27.99 ±4.31 0.799 R1, linea i y 1.80 ±0.11 1.47 ±0.24 0.096 1.18 ±0.12 1.11 ±0.11 0.498 R2, RG con ibu ion 0.03 ±0.00 0.03 ±0.00 >0.999 0.04 ±0.00 0.01 ±0.00 <0.001 R3, leng h o side chains 11.48 ±0.17 11.01 ±1.57 0.633 11.93 ±0.40 38.59 ±5.97 0.002 DM (%) e 58.67 ±2.08 54.67 ±1.53 0.055 61.33 ±1.53 59.00 ±0.33 0.061 Da a a e exp essed as he mean ±SD (n=3). a Based on AIR. b Exp essed as g kg −1 o d y ex ac . c Rela i e %. d Mola a ios: exhibi ing he p ima y s uc u al p ope ies o pec in molecules; HG =GalA –Rha, homogalac u onan; RGI =2(Rha) + A a + Gal, R1 =GalA/(Fuc + Rha + Glu + A a + Gal+Xyl), he linea i y o pec in; R2 =Rha/GalA, he con ibu ion o RG o pec in popula ion; R3 =(Gal+A a)/ Rha, he leng h o side chains a ached o RGI. e Deg ee o me hyl es e ifica ion (DM) de e mined by ATR/FT- IR. Cha ac e iza ion o pec in om by-p oduc s o he oli e oil indus y www.soci.o g J Sci Food Ag ic 2025 © 2025 The Au ho (s). Jou nal o he Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y. wileyonlinelib a y.com/js a 3 10970010, 0, Downloaded om h ps://scijou nals.onlinelib a y.wiley.com/doi/10.1002/js a.14316 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [17/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License aw ma e ial be o e ci ic acid ex ac ion was c ucial o p e en pec inoly ic enzyme ac i i y. The pec in ex ac ion yield anged om 24.8 g kg −1 (aeOMSbP) o 48.2 g kg −1 (mmOMSbP). A signi - ican ly highe yield was ob ained om mmOMSbP (48.2 g kg −1 ) compa ed o meOMSbP (40.0 g kg −1 ), ep esen ing a 20% dec ease in ex ac ion e ficiency a an ea lie ipeness s age. How- e e , hese yield we e lowe han hose epo ed o pec in om o ange peel (app oxima ely 200 g kg −1 ), which is he main ag oindus ial was e used o he comme cial p oduc ion o pec in, 25 pomeg ana e peel (110 g kg −1 ), 26 young apples (80 g kg −1 ) 27 and suga bee (160 g kg −1 ). 28 The p o ein con en o he ex ac ed pec in was wi hin he ange epo ed o com- me cial apple pec in (app oxima ely 30 g kg −1 ) 29 and highe han ha ound in co ee pulp (app oxima ely 10 g kg −1 ). 30 Addi ion- ally, he phenolic con en o OMSbP pec in was highe han ha o ci us pec in (1.5–1.8 g kg −1 ), bu he simila o alues epo ed o apple pec in (up o 6 g kg −1 ). 31 To assess he ca bohyd a e composi ion and deg ee o b anch- ing, he monosaccha ide con en was de e mined using gas ch o- ma og aphy. Significan di e ences we e obse ed be ween di e en ipeness s ages, wi h ea ly-ha es ed eOMSbP samples showing a highe con en o bo h u onic acids and neu al suga s. This esul ed in a p e e en ial loss o o al ca bohyd a es om he polyme ic cell wall in la e-ha es ed samples, pa icula ly Rha and Xyl, which con ibu e o a abinoxylan s uc u es in he cell wall. As shown in Table 1, OMSbP samples con ained se en p ima y monosaccha ides: Rha, A a, Gal, Glu, Xyl, Fuc and Man, in a ying p opo ions. GalA was he dominan monosaccha ide, wi h le els anging om 500.3 g kg −1 (mmOMSbP) o 624.8 g kg −1 (aeOMSbP), confi ming i s high HG con en . The GalA a io exceeded 50% in all analyzed pec in samples, which is highe han he 35% epo ed by p e ious s udies. 32 Rha, Gal and A a oge he accoun ed o o e hal o he o al neu- al suga s, sugges ing a high con ibu ion o RG-I o he pec in s uc u e. The HG con en (49.34–60.43%) was g ea e han ha o RG-I (21.45–27.99%), indica ing ha he linea backbone s uc- u e p edomina ed o e side chains. Pec in con aining mo e han 74% GalA has been epo ed o influence gu mic obio a compo- si ion and i s mic obial me aboli es, 33 pa icula ly h ough he ac i i y o RG-I, which plays a ole in modula ing immune esponse. 34 In he p esen s udy, he R1 alues o OMSbP pec in samples (1.11–1.80) we e ound lowe han hose epo ed o g ape ui peel pec in (2.52) 35 and pis achio g een hull pec in (1.97), 36 indica ing ha OMSbP pec in has a highe deg ee o b anching. Howe e , he R1 alue o aeOMSbP pec in (1.80) was highe han o swee lemon peel (1.53), 37 highligh ing a ia ions in s uc u al complexi y among di e en sou ces. The R2 and R3 alues u he suppo ed ha he majo con ibu ions o pec in s uc u e we e om RG-I domains, wi h mmOMSbP exhibi ing he highes deg ee o b aching (38.59) compa ed o o he sam- ples (app oxima ely 11). To be e unde s and he ela ionship be ween he s uc u al componen s and pec in p ope ies, Pea son co ela ion coe fi- cien s we e calcula ed (Fig. 1). S ong co ela ions (>0.95) we e obse ed be ween mul iple a iables, including yield and o al ca bohyd a es (−0.96), yield and a abinose (0.97), yield and man- nose (0.99), yield and R1 (−0.97), and yield and DPPH (−0.98). Addi ionally, significan co ela ions we e ound be ween p o ein con en and galac ose (−0.96), phenols and xylose (0.95), o al ca bohyd a es and galac u onic acid (0.98), o al ca bohyd a e and mannose (−0.98), o al ca bohyd a e and glucose (0.96), o al ca bohyd a e and DPPH (0.99), hamnose and R2 (0.99), hamnose and R3 (−0.96), xylose and R2 (0.95), mannose and R1 (−0.97), glu- cose and hamnogalac u onan I (0.99), glucose and R1 (−0.96), homogalac u onan and DPPH (0.96), and R1 and DPPH (0.97). The s uc u al complexi y and unc ionali y o pec in a e closely ela ed. 38 Recen s udies ha e modeled he co ela ion be ween pec in s uc u e and i s echno- unc ional p ope ies. Xie e al. 39 es ablished ela ionships be ween pec in cha ac e is ics and unc ional a ibu es using Pea son co ela ion coe ficien s, highligh ing p ope ies such as heology, chewiness, adhesion, eeze– haw s abili y and ha dness. These esul s ein o ced he no ion ha pec in con o ma ion, monosaccha ide composi ion, and molecula weigh a e he p ima y ac o s influencing i s phys- icochemical beha io . Fu he s udies ha e demons a ed he impac o pec in s uc u e and i s DM on gela iniza ion p ope - ies. 40 A s udy by Luo e al. 41 alida ed a mul iple eg ession model demons a ing how a ia ions in GalA con en , molecula weigh , DM and amida ion deg ee influence gela iniza ion cha - ac e is ics such as peak and alley iscosi y, as well as se back and b eakdown alues. These findings unde sco e he signifi- cance o s uc u al pa ame e s in de e mining pec in unc ional- i y, u he suppo ing he complex in e play be ween chemical composi ion and echno- unc ional a ibu es. An ioxidan ac i i y and co ela ion wi h s uc u al componen s Figu e 2A p esen s he an ioxidan ac i i y o he ex ac ed pec in, measu ed h ough DPPH adical sca enging. The EC 50 alues indi- ca e ha meOMSbP exhibi ed a 6% highe ee adical sca enging abili y compa ed o mmOMSbP. This inc ease is di ec ly p opo - ional o he 7% highe phenolic con en ound in meOMSbP, con- fi ming ha he an ioxidan ac i i y o hese ex ac s is p ima ily a ibu ed o hei phenolic compounds a he han he pec in s uc u e i sel . Phenols eadily bind o suga s, ans e ing hei an ioxidan p ope ies o hese molecules. 7 The EC 50 alues ob ained o he ex ac ed pec in de i ed om oli e pomace by-p oduc s we e lowe han hose epo ed o Spi ulina sp., a mic oalga known o i s no able an ioxidan capaci y. The mos e ec i e concen a ion was 2.5 mg mL −1 , and his ac i i y has been mainly a ibu ed o he p esence o bioac i e compounds such as phenolic acids, phycocyanins and ocophe ols. 42 Suga s can also enhance he bioa ailabili y o phenols, c ea ing a syne gis ic unc ional ela ionship be ween he wo compound g oups. 43 This in e ac ion is o high nu i ional in e es because i makes his ex ac ed pec in om OMSbP mo e aluable o he o mula ion o heal h-o ien ed ood p oduc s compa ed o com- me cial pec in ha lack phenolic associa ions. To u he in es iga e whe he specific cha ac e is ics o he OMSbP-de i ed pec in, such as yield, chemical composi ion and molecula s uc u e, influence i s an ioxidan capaci y, Pea son co ela ion coe ficien s we e calcula ed. Figu e 2B demons a es ha DPPH exhibi ed s ong co ela ions (>±0.5) wi h mul iple a iables, bo h posi i e and nega i e. The s onges co ela ion was ound be ween DPPH and mannose con en ( =−0.9967). To assess he significance o his ela ionship, a simple linea eg ession model was applied, yielding he equa ion y=2.3805–0.1991x, whe e ‘y’ ep esen s an ioxidan ac i i y and ‘x’ ep esen s mannose concen a ion. The model confi med ha mannose had a significan impac (P<0.05) on DPPH, explaining 99% o he a iance in he da a (R 2 =0.9901). This sugges ha inc easing mannose con en in ex ac ed pec in could enhance hei an ioxidan p ope ies. P e ious s udies ha e shown ha he an ioxidan ac i i y o oligogalac u onides is closely ela ed www.soci.o g A Fe nandez-P io e al. wileyonlinelib a y.com/js a © 2025 The Au ho (s). Jou nal o he Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y. J Sci Food Ag ic 2025 4 10970010, 0, Downloaded om h ps://scijou nals.onlinelib a y.wiley.com/doi/10.1002/js a.14316 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [17/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License o hei s uc u al a ibu es, including monosaccha ide composi- ions and molecula weigh . 44-47 The an ioxidan mechanisms o pec in a e in ica e, and u he esea ch is needed o explo e he ela ionship be ween polysaccha ide s uc u e and an ioxi- dan unc ion in mo e dep h. 48 Deg ee o me hyl es e ifica ion o ex ac ed pec in om OMSbP The ATR/FT-IR spec a we e analyzed o de e mine whe he he lyophilized OMSbP pec in ex ac ed om di e en oli e a ie ies and ipeness s ages a ec ed unc ional g oups and bonding con- figu a ions (Fig. 3). O e all, sligh a ia ions we e obse ed in he spec al p ofiles o ex ac ed pec in om he A bequina a ie y (aeOMSbP) (Fig. 3A, g een line) and he Hojiblanca a ie y (heOMSbP) (Fig. 3A, o ange line). Addi ionally, di e ences we e no ed be ween ea ly-season (meOMSbP) (Fig. 3B, pink line) and la e- season (mmOMSbP) (Fig. 3B, blue line) pec in samples. The b oad abso p ion band a 3400 cm −1 , co esponding o O H s e ching ib a ion o hyd oxyl g oups, appea ed sha pe in mmOMSbP compa ed o meOMSbP, likely because o a lowe phenolic con- en (Table 1). 49 No significan di e ences we e de ec ed be ween he wo oli e a ie ies, aligning wi h p ecious findings, and po en- ially a ibu ed o he high dissocia ion cons an o he ci ic acid used du ing ex ac ion. 50 The abso p ion band a 2925 cm −1 a ibu ed o C H(—CH, —CH 2 ,—CH 3 ), emained consis en ac oss all pec in samples, sugges ing ha ipeness did no influence hese unc ional g oups. Howe e , aeOMSbP exhibi ed a highe in ensi y in his egion compa ed o heOMSbP. No ably, ex ac ed pec in om la e-season (mmOMSbP) and Hojiblanca oli es (heOMSbP) dis- played a dec ease in he band nea 1730 cm −1 , co esponding o C O s e ching o he me hyles e ified ca boxyl g oups (COO-R), whe eas he e was an inc ease a 1620 cm −1 , associa ed wi h ca boxyla e ion (COO − ) s e ching, indica ing sligh a ia- ions in he deg ee o DM, as eflec ed in Table 1. The abso p ion bands a 1146 cm −1 and 1405 cm −1 , associa ed wi h C—OH s e ching in side g oups and C—O—C glycosidic bond ib a ion, espec i ely, emained unchanged ac oss all samples, indica ing consis ency in glycosidic linkages. 51,52 Simila ly, cha ac e is ic bands a 1250 cm −1 ,(—CH 3 CO s e ching), 957 cm −1 (C O bend- ing), 912 cm −1 ( ocking mode o CH 3 ) and 820 cm −1 (—CCH and —COH bending a he C-6 posi ion) we e main ained, ega dless o ipeness s age o oli e a ie y, ein o cing he s uc u al s abil- i y o ex ac ed pec in. 1 H NMR analysis The 1 H NMR spec a (Fig. 4) o he ex ac ed pec in om meOMSbP and mmOMSbP we e acqui ed a 500 MHz o u he Figu e 1. Pea son co ela ion coe ficien hea map showing ela ionships among s uc u al, con o ma ional cha ac e is ics and an ioxidan capaci y o pec in. The colo g adien ep esen s he Pea son co ela ion coe ficien , wi h alues displayed wi hin each squa e. Cha ac e iza ion o pec in om by-p oduc s o he oli e oil indus y www.soci.o g J Sci Food Ag ic 2025 © 2025 The Au ho (s). Jou nal o he Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y. wileyonlinelib a y.com/js a 5 10970010, 0, Downloaded om h ps://scijou nals.onlinelib a y.wiley.com/doi/10.1002/js a.14316 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [17/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License co obo a e and cla i y hei s uc u es ea u es. Spec a we e eco ded a 60 °C using samples dissol ed in D 2 O, aiming o educe sample iscosi y. Measu ing a his empe a u e also had he added benefi o shi ing he esidual wa e signal o app ox- ima ely 4.4 ppm, a oiding o e lap wi h ele an pec in signals. The mos ele an signals appea in he 3.0–5.5 ppm egion, co - esponding o he skele al and anome ic p o ons o he monosaccha ides. Pec in samples wi h a high deg ee o me hyla- ion, such as hose analyzed, ypically display anome ic p o on peaks a ound 5 ppm. In pa icula , anome ic H-1 and H-5 (COO − ) p o ons a e obse ed a ⊐5.3 ppm and ⊐4.8 ppm, espec- i ely, cha ac e is ics o GalA esidues linked ia ⊍-1,4 glycosidic bonds. Addi ionally, a peak a 4.6 ppm co esponds o H-4 p o- ons, whe eas signals a app oxima ely 4.2 and 4.1 ppm a e Figu e 2. (A) DPPH adical sca enging ac i i y (EC 50 ) o pec in samples. Values (mg mL −1 ) a e exp essed as he mean ±SD (n=3). S a is ical signi - icance be ween a ie ies and ipeness s ages is indica ed by P- alue. (B) Co ela ion analysis be ween chemical composi ion and an ioxidan capaci y (EC 50 ,mgmL −1 ). The ba leng h ep esen s he Pea son co ela ion coe ficien , wi h alues displayed wi h he ba s. www.soci.o g A Fe nandez-P io e al. wileyonlinelib a y.com/js a © 2025 The Au ho (s). Jou nal o he Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y. J Sci Food Ag ic 2025 6 10970010, 0, Downloaded om h ps://scijou nals.onlinelib a y.wiley.com/doi/10.1002/js a.14316 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [17/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License Figu e 3. Fou ie ans o m-in a ed spec oscopy (ATR/FT-IR) spec a o lyophilized OMSbP pec in ex ac ed om (A) di e en oli e a ie ies and (B) di e en ipeness s ages. A bequina a ie y (aeOMSbP, g een line), hojiblanca a ie y (heOMSbP, o ange line), ea ly-s age ipeness (meOMSbP, pink line) and ma u e-s age ipeness (mmOMSbP, blue line). Figu e 4. 1D 1 HNMR spec a o ex ac ed pec in om meOMSbP and mmOMSbP eco ded a 500 MHz in D₂Oa 60°C. Cha ac e iza ion o pec in om by-p oduc s o he oli e oil indus y www.soci.o g J Sci Food Ag ic 2025 © 2025 The Au ho (s). Jou nal o he Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y. wileyonlinelib a y.com/js a 7 10970010, 0, Downloaded om h ps://scijou nals.onlinelib a y.wiley.com/doi/10.1002/js a.14316 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [17/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License a ibu ed o H-3 and H-2 p o ons, espec i ely, in ag eemen wi h a p e ious s udy. 53 The egion be ween 3.0 o 4 ppm shows o e - lapping signals assigned o galac ose, a abinose and hamnose ing p o ons, sugges ing he p esence o a abinogalac an s uc- u es o pec ic side chains. Mo eo e , signals a ound 1.0–1.5 ppm a e likely associa ed wi h he me hyl g oups o hamnose, indica- i e o b anched egions wi hin he pec in s uc u e. No ably, a se ies o double o double s o double s (ddd) signals obse ed be ween ⊐2.7 and ⊐2.9 ppm may be ela ed o he binding o pec in o he ci ic acid used du ing ex ac ion. These signals a e a ibu ed o me hylene p o ons o ci ic acid, as p e iously sug- ges ed by Ke mani e al. 54 This spec al p ofile confi ms he com- plex composi ion o he ex ac ed pec in, which comp ise a GalA backbone wi h po en ial subs i u ions by neu al suga . Su ace mo phology analysis by SEM The su ace mo phology o lyophilized OMSbP pec in ex ac ed om di e en oli e a ie ies (Fig. 5A–D) and a di e en ipeness s ages (Fig. 5E–H) was analyzed using SEM (Fig. 5). The SEM images e ealed s uc u al di e ences be ween pec in samples. In he eOMSbP samples, deg ada ion in o smalle agmen s was obse ed a e he ha es season, leading o a educ ion in he pa icle size unde iden ical imaging condi ions (Mag =50×and AV =2.0 kV/Mag =250×and AV =2.0 kV). Ex ac ed pec in om aeOMSbP exhibi ed a smoo h su ace, likely because o i s highe HG con en and ewe s uc u al dis up ions. By con as , mOMSbP pec in displayed a dis inc s uc u e wi h isible long c acks on he su ace. This he e ogeneous a chi ec u e, cha ac e - ized by ough, compac and flaky s uc u es, may be a ibu ed o a lowe con en o acidic suga side chains, which influences he o ganiza ion and mechanical p ope ies o he ex ac ed pec in. 23 The mal s abili y o OMSbP pec in-based films TGA and i s de i a i e (DTGA) cu es (Fig. 6A,B) p o ide c i ical insigh s in o he he mal s abili y and decomposi ion beha io o pec in-based films de i ed om OMSbP, specifically om di - e en ipeness s ages. Unde s anding hese he mal ansi ions is essen ial no only o e alua ing p ocessabili y unde hea , bu also o p edic ing hei beha io in end-use applica ions such as ood packaging, edible films o con olled- elease sys ems. As shown in Fig. 6A, bo h pec in samples exhibi ed a simila he - mal deg ada ion p ofile, wi h mino di e ences mo e e iden in he DTGA cu es (Fig. 6B). The deg ada ion ollowed a ypical h ee-s age pa e n, cha ac e is ic o polysaccha ide-based ma e ials. 55,56 The ini ial mass loss, obse ed a ound 93 °C, co - esponded o he e apo a ion o ee and bound mois u e, eflec ing he hyg oscopic na u e o pec in and he p esence o hyd oxyl g oups capable o o ming hyd ogen bonds wi h wa e . 35 This s ep is pa icula ly impo an in edible films because mois u e e en ion can a ec mechanical in eg i y and shel -li e pe o mance. The second deg ada ion s age, occu ing a app oxima ely 233 °C, was associa ed wi h he depolyme iza- ion and b eakdown o he HG backbone, which is mainly com- posed o GalA uni s. Addi ional decomposi ion p ocessed we e ela ed o he clea age o glycosidic linkages and he deca box- yla ion o GalA. The mal esis ance in his phase may be influ- enced by he deg ee o es e ifica ion, molecula weigh and p esence o impu i ies o esidual phenolic compounds co- ex ac ed om he oli e by-p oduc ma ix. 57,58 A hi d majo decomposi ion peak, obse ed a ound 443 °C, likely co e- sponds o he b eakdown o mo e he mally s able ac ions and he oxida i e deg ada ion o ca bonaceous esidues. The p esence o a mino shoulde in he DTGA cu es sugges s pa - ial ca boniza ion and esidual cha o ma ion, po en ially as a esul o lignocellulosic agmen s o phenolic esidues com- monly e ained in ex ac ed pec in om ag o-indus ial esi- dues. 58 These findings indica e ha OMSbP pec in exhibi he mal s abili y compa able o ha o comme cial pec in, sup- po ing i s po en ial use in he mally p ocessed applica ions such as low- empe a u e d ying, ex usion o lamina ion. 59 CONCLUSIONS The p esen s udy highligh s he po en ial o ex ac ed pec in om OMSbP as a unc ional and sus ainable ing edien o ood applica ions. Pec in ob ained om ea ly-s age ipeness OMSbP samples (eOMSbP) exhibi ed a highe GalA con en and deg ee o me hyla ion, along wi h dis inc monosaccha ide composi ion di e ences. Despi e chemical a ia ions among ex ac ed pec in om di e en oli e a ie ies and ipeness s ages, hey displayed significan an ioxidan p ope ies, likely a ibu ed o hei pheno- lic con en and a s ong co ela ion wi h mannose concen a ion. The he mal s abili y o OMSbP pec in-based films was compa a- ble o ha o comme cial pec in, mee ing key heological Figu e 5. Su ace mo phology o lyophilized OMSbP pec in ex ac ed om di e en oli e a ie ies and di e en ipeness s ages. (A, B) A bequina a ie y (aeOMSbP), (C, D) Hojiblanca a ie y (heOMSbP), (E, F) ea ly-s age ipeness (meOMSbP) and (G, H) ma u e-s age ipeness (mmOMSbP), analyzed by SEM a wo magnifica ions (Mag.). Uppe SEM images we e aken a Mag =50x and AV =2.0 kV (scale ba =100 μm), while lowe SEM images we e aken a Mag =250x and AV =2.0 kV (scale ba =30 μm). www.soci.o g A Fe nandez-P io e al. wileyonlinelib a y.com/js a © 2025 The Au ho (s). Jou nal o he Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y. J Sci Food Ag ic 2025 8 10970010, 0, Downloaded om h ps://scijou nals.onlinelib a y.wiley.com/doi/10.1002/js a.14316 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [17/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License p ope ies equi ed in he ood indus y. The applica ion o he - mal ea men , simila o hose being inc easingly adop ed by oli e oil pomace p ocesso s, combined wi h ci ic acid ex ac ion, enables he p oduc ion o bioac i e pec in wi h film- o ming and hea - esis an p ope ies. These cha ac e is ics suppo hei po en ial use in ood p ese a ion and o mula ion. The findings o he p esen s udy confi m ha he mal p e- ea men s a e an e ec i e s a egy o enhancing he ex ac ion o bioac i e com- pounds om oli e pomace, con ibu ing o he ull alo iza ion o his by-p oduc . The pec in ob ained demons a ed s uc u al in eg i y, unc ional s abili y and an ioxidan ac i i y, ein o cing i s sui abili y o inco po a ion in o ood p oduc s a he same ime as p omo ing sus ainabili y in he oli e oil indus y. Fu u e esea ch should ocus on op imizing ex ac ion condi ions o u - he enhance pec in yield and bioac i i y. Addi ionally, in es iga - ing he bioa ailabili y and heal h e ec s o OMSbP-de i ed pec in in unc ional ood applica ions could expand i s po en ial in nu a- ceu icals and clean-label o mula ions. AUTHOR CONTRIBUTIONS SM-dlP was esponsible o s udy concep ualiza ion. AF-P, LB-C, CL-dD and AV-L we e esponsible o he me hodology. AF-P, EM-P and SM-dlP we e esponsible o o mal analysis. AF-P, CMM-L and SM-dlP we e esponsible o in es iga ions. AF-P and SM-dlP we e esponsible o esou ces. AF-P and SM-dlP we e esponsible o w i ing he o iginal d a . AF-P and SM-dlP we e esponsible o e iewing and edi ing. AF-P and SM-dlP we e esponsible o supe ision. SM-dlP was esponsible o unding acquisi ion. All au ho s ha e ead and app o ed he final e sion o he manusc ip submi ed o publica ion. FUNDING This s udy was suppo ed by he TED2021-130521A-I00 esea ch p ojec (Minis y o Science and Inno a ion, Go e nmen o Spain) in o he Reco e y, T ans o ma ion, and Resilience Plan unding by Nex Gene a ionEU. ACKNOWLEDGEMENTS A ica Fe nandez-P io has he benefi o a Juan de la Cie a pos - doc o al ellowship FJC2021-047485-I om he Spanish Minis y o Science and Inno a ion. Luna Ba e a-Chamo o has he ben- efi o a PIF doc o al ellowship suppo ed by he VII P og am o Inne Ini ia i e o Resea ch and T ans e o Uni e si y o Se ille (VII-PPIT-US). El i a Ma quez-Pa adas has he benefi o a FPU doc o al ellowship (FPU22/01097) om he Spanish Minis y o Science and Inno a ion. C. López de Dicas illo acknowledges he awa d o he Spanish go e nmen MCIU/AEI o he IATA-CSIC as Cen e o Excellence Acc edi a ion Se e o Ochoa (CEX2021- 001189- S/MCIU/AEI/10.13039/501100011033). CONFLICTS OF INTEREST The au ho s decla e ha hey ha e no conflic s o in e es . DATA AVAILABILITY Da a a e a ailable om he co esponding au ho upon eason- able eques . REFERENCES 1 Reichembach LH, Gue e o P, de Oli ei a Pe kowicz CL and de la Caba K, Valo iza ion o pec ins om co ee was es o he de elop- men o pec in-chi osan films. Ca bohyd Polym 334:122057 (2024). h ps://doi.o g/10.1016/j.ca bpol.2024.122057. 2 Lama-Muñoz A, Rubio-Senen F, Be múdez-O ia A, Fe nández- Bolaños J, P io ÁF and Rod íguez-Gu ié ez G, The use o indus ial he mal echniques o imp o e he bioac i e compounds ex ac ion and he oli e oil solid was e u iliza ion. Inno Food Sci Eme g Technol 55:11–17 (2019). h ps://doi.o g/10.1016/j.i se .2019.05.009. 3 Sánchez-Sánchez C, González-González A, Cuad os-Salcedo F and Cuad os-Blázquez F, Two-phase oli e mill was e: a ci cula economy solu ion o an imminen p oblem in sou he n Eu ope. J Clean P od 274:274 (2020). h ps://doi.o g/10.1016/j.jclep o.2020.122789. 4 Fe nández-P io MÁ, Cha fiA, Be múdez-O ia A, Rod íguez-Juan E, Fe nández-Bolaños J and Rod íguez-Gu ié ez G, Deep eu ec ic sol- en s imp o e he bio efine y o alpe ujo by ex ac ion o bioac i e molecules in combina ion wi h indus ial he mal ea men s. Food Biop od P ocess 121:131–142 (2020). h ps://doi.o g/10.1016/j. bp. 2020.02.001. Figu e 6. (A) The mog a ime ic analysis (TGA) cu es and (B) hei co esponding de i a i e TGA (DTGA) o OMSbP pec in-based films ex ac ed om ea ly-s age ipeness (meOMSbP, pink line) and ma u e-s age ipeness (mmOMSbP, blue line). Cha ac e iza ion o pec in om by-p oduc s o he oli e oil indus y www.soci.o g J Sci Food Ag ic 2025 © 2025 The Au ho (s). Jou nal o he Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y. wileyonlinelib a y.com/js a 9 10970010, 0, Downloaded om h ps://scijou nals.onlinelib a y.wiley.com/doi/10.1002/js a.14316 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [17/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License