Influence of Irrigation Modalities (Irrigation Management and Dryland), Fruit Ripening, and Cultivation Modality (Organic and Conventional) on Quality and Chemosensory Profile of Hojiblanca and Picual Extra Virgin Olive Oils

RESEARCH ARTICLE www.ejls .com Influence o I iga ion Modali ies (I iga ion Managemen and D yland), F ui Ripening, and Cul i a ion Modali y (O ganic and Con en ional) on Quali y and Chemosenso y P ofile o Hojiblanca and Picual Ex a Vi gin Oli e Oils Rocio Ríos-Reina, F ancisco Camacho, M. Lou des Mo ales, B ígida Jiménez-He e a, and Raquel M. Callejón* A s udy wi h con olled field and au hen ic samples o oli es, ob ained in simila condi ions o soil, clima e, egion, ha es , and wi h he same cul i a ion echniques and conside ing simul aneously diffe en ag onomic ac o s (oli e a ie y, ui ipening deg ee, i iga ion, and o ganic o con en ional p oduc ion sys em) is pe o med o e alua e hei influence on quali y and added alue o ex a i gin oli e oil (EVOO). Ag onomical and physicochemical pa ame e s, polyphenols, ocophe ols, and a y acid composi ion and ola ile and senso y p ofiles a e de e mined in Hojiblanca and Picual VOOs ob ained om diffe en ui ipening deg ees and diffe en cul i a ion modali ies (con en ional wi h and wi hou i iga ion, and o ganic wi h i iga ion). Among ola ile compounds, 1-hyd oxy-2-p opanone, (E)-linalool oxide, and 2-ace yl u an a e desc ibed o he fi s ime in EVOO. The a iable ha mos influences he chemosenso y composi ion o EVOOs is he a ie y, ollowed by he s age o ipeness, and, wi hin each a ie y, he cul i a ion modali y. O ganic i iga ion diffe om con en ional modali ies, showing significan diffe ences in acidi y, s abili y, ocophe ol and polyphenol con en s, a y acid composi ion, and senso y a ibu es. P ac ical Applica ions: Resul s a e o g ea impo ance, due o hei applicabili y o he EVOO sec o , allowing one o know he quali a i e, chemical and o ganolep ic diffe ences be ween o ganic and con en ional EVOO, and ac o s ha imp o e he quali y and pe o mance o EVOO. 1. In oduc ion Vi gin oli e oil (VOO) p oduc ion is mainly concen a ed in Medi e anean coun ies, Spain being he leade in bo h oli e D . R. Ríos-Reina, F. Camacho, P o . M. L. Mo ales, D . R. M. Callejón Á ea de Nu ición y B oma ología, Dp o. de Nu ición y B oma ología, Toxicología y Medicina Legal Facul ad de Fa macia, Uni e sidad de Se illa C/P. Ga cía González n°2, Se illa E-41012, Spain E-mail: [email p o ec ed] D . B. Jiménez-He e a Depa men o Technology, Pos ha es , and Food Indus ies, IFAPA “Cab a” Cen e Ins i u e o Resea ch and T aining in Ag icul u e and Fishe ies C a. Cab a-Doña Mencía, KM. 2.5, Cab a, Có doba 14940, Spain DOI: 10.1002/ejl .202000375 g o e su ace and oli e oil p oduc ion.[1] VOO, and especially ex a i gin oli e oil (EVOO), is a p oduc ha is inc easingly ap- p ecia ed by consume s o i s o ganolep ic, nu i ional cha ac e is ics, and high quali y and benefi s o human heal h.[2,3] The Eu opean Union, he Codex Alimen- a ius and he In e na ional Oli e Council ha e diffe en egula ions o classi y oli e oil acco ding o hei quali y.[4] As is well known, lipid composi ion is one o he main con ibu o s o nu i ional and unc- ional quali y, due o he high p opo ion o monounsa u a ed a y acids (MUFA) o oli e oil. Polyphenols also ha e an impo - an ole in quali y since hey a e espon- sible o o ganolep ic cha ac e is ics, s abil- i y, and heal h benefi s. In ac , oli e oil is he only ood p oduc ha has an au ho- ized heal h claim on polyphenols:[5] “oli e oil polyphenols con ibu e o he p o ec ion o blood lipids om oxida i e s ess.” Mo e- o e , oli e oil no only has a heal h claim on oli e oil polyphenols, i also has ano he on unsa u a ed a y acids. O he essen ial compounds in oli e oil quali y a e ola ile compounds since hey con ibu e g ea ly o he o e all a oma and fla o .[6–8] Quan i a i e and quali a i e chemical composi ion o VOO is affec ed by many ac o s such as ag onomical and echnical ac- o s, ha ing a g ea impac on i s quali y. They mainly include he gene ic ac o such as oli e a ie y; he en i onmen al ac- o s, such as pedoclima ic ac o ela ed o he geog aphical a ea, ma u a ion deg ee o he ui , i iga ion, c op season o he ag o- nomical p ac ices used (e.g., con en ional o o ganic p ac ices); and echnological ac o s, such as he condi ions o oil ex ac ion p ocess.[9–13] Al hough p e ious s udies ha e shown ha he oli e a ie y and ui ipening s age ha e a g ea e effec on nu i ional and physical-chemical pa ame e s han he cul i a ion modali y,[14] i - iga ion seems o influence he composi ion and o ganolep ic cha ac e is ics o oli e oil [15] al hough i s le el on VOO quali y is no ye clea and is s ill con o e sial.[16–20] Eu . J. Lipid Sci. Technol. 2021,123, 2000375 © 2021 Wiley-VCH GmbH 2000375 (1 o 11) www.ad ancedsciencenews.com www.ejls .com Ano he pa icula ly impo an ac o o conside is he ype o a ming, due o he inc ease in demand o o ganic p oduc s by consume s and he s ic egula ion o he applica ion o ce ain chemical p oduc s o c ops, such as insec icides, ungicides, o he bicides.[21] Howe e , despi e he inc ease in he su ace a ea in o ganic ag icul u e oli e g o es in Spain, and he po- en ial adul e a ions and auds in his kind o p oduc , he e is s ill a lo o igno ance abou how he ype o c op, o ganic o con en ional, affec s chemical composi ion since o da e, ew compa a i e s udies ha e been pe o med be ween o ganic and con en ional oli e oils.[10,14,22,23–25] Indeed, in some cases, such s udies ha e ob ained con adic o y and inconclusi e esul s due o he expe imen al design no conside ing ac o s such as a i- e y, yea o ha es , place o p oduc ion, same soil ype, ipeness o he ui a he ime o ha es , he same cul i a ion ech- niques, e c. In o he wo ds, hese s udies we e no pe o med using con olled and compa able samples ob ained unde he same condi ions in which he only a iable was he c op ype. The e o e, a s udy wi h con olled field samples, ob ained un- de he same condi ions, conside ing simul aneously diffe en ag onomic ac o s (oli e a ie y, ui ipening deg ee, i iga ion, and o ganic o con en ional) would ep esen an ad ance in he knowledge on EVOO, p e en ing aud and p o iding in o ma- ion o he indus y, p oduce s, and consume s, on he quali y and added alue o hese p oduc s. In his con ex , he aim o his wo k is o s udy simul ane- ously he influence o i iga ion and p oduc ion sys em (o ganic and con en ional) on he quali y o EVOOs (including o al phe- nols, ocophe ols, a y acid composi ion, and ola ile and senso y p ofile) om Hojiblanca and Picual a ie ies and diffe en ui ipening deg ee, ob ained unde simila condi ions. 2. Expe imen al Sec ion 2.1. Chemicals The analy ical s anda ds o a y acids, ocophe ols, ola ile, and phenolic compounds we e o high pu i y (>95%) and ob ained om comme cial sou ces (Me ck, Da ms ad , Ge many; Sigma- Ald ich, Mad id, Spain; Fluka, Mad id, Spain). Wa e was ob- ained om a Milli-Q pu ifica ion sys em (Millipo e, USA). A se ies o C10 o C40 s aigh -chain n-alkanes (50 mg L−1in n- hexane) pu chased om Fluka was used o calcula e linea e en- ion index (LRI). All he sol en s used we e o analy ical o HPLC g ade (Sigma-Ald ich). 2.2. Oli e F ui Samples Oli e ui s we e collec ed om oli e ees o wo o he a ie ies mos cul i a ed in Andalusia, he Picual and Hojiblanca a ie ies, cul i a ed in h ee modali ies: con en ional wi h i iga ion a m- ing (CI), o ganic wi h i iga ion a ming (OI) and con en ional wi hou i iga ion o d yland a ming (CD); and we e ha es ed a ou diffe en s ages o ipening. The oli e ees cul i a ed by all h ee modali ies we e loca ed in wo o cha ds, 17 km apa om one o he , a simila al i udes (app oxima ely 300 m abo e sea le el) and wi h simila wea he condi ions. Hence, bo h a - eas ha e a con inen al Medi e anean clima e wi h d y summe s and mild win e . Rain all occu s om au umn o sp ing, wi h a mean annual ain all o 400 mm. The a e age annual empe a- u e is 17 °C, eaching -1.3 °C in win e and 43 °C in summe . Soils ha e low dep hs, being mainly o e limes one and siliceous s ones. They ha e a loamy ex u e and an alkaline pH (in he 8–8.5 ange). Thus, he oli e ees cul i a ed wi h con en ional modal- i y (d yland and i iga ion a ming) we e loca ed a he Ag icul- u al Resea ch T aining Cen e in “Cab a” in he p o ince o Co - doba, Spain, while oli e ees cul i a ed wi h o ganic modali y (i iga ion a ming) we e si ua ed in “Baena” in he p o ince o Co doba, Spain. The i iga ion modali ies we e pe o med using a localized i iga ion sys em wi h ou d ip nozzles wi h an appli- ca ion o 40 L pe day o each ee, deli e ing a o al o 1500 m3 pe hec a e. Ten oli e ees we e andomly selec ed (always among he mos loaded o gua an ee sampling) o he Picual a ie y and con- en ional i iga ion modali y, 10 ees o he Picual a ie y and con en ional d yland modali y, and 10 ees o he Picual a ie y and o ganic i iga ion modali y. The same selec ion was ca ied ou o oli e ees o he Hojiblanca a ie y (10 con en ional wi h d yland, 10 con en ional wi h i iga ion, and 10 o ganic wi h i - iga ion). Oli e sampling was done by andomly hand-picking only heal hy ui s (wi hou any ype o in ec ion o physical damage), e e y 2 weeks, beginning in Oc obe 2016 and ending in Decem- be 2016, and comp ising diffe en ha es pe iods. I con o ms a o al o 4 ipening s ages: 22 om Oc obe (phase I), 3 om No embe (phase II), 19 om No embe (phase III) and 2 om Decembe (phase IV). The ipening index (RI) o each ha es was de e mined ac- co ding o he me hodology p oposed by Uceda and F ias.[26] The ui RI alues o each modali y a e shown in Table S1 (Suppo - ing In o ma ion). 2.3. Ag onomical Indices 2.3.1. Weigh o 100 F ui s The ui weigh was de e mined as he weigh o 100 d upes an- domly picked om aliquo s o samples ha had been p e iously homogenized. 2.3.2. Resis ance o De achmen (RD) This pa ame e was e alua ed using a dynamome e (Co ex, Swi ze land) measu ing he o ce (in g ams) necessa y o b eak he ui peduncle and de ach i om he b anch. 2.3.3. Oil Con en (CAH and CAS) and Humidi y Con en The sample humidi y con en was de e mined acco ding o he UNE S anda d Spanish me hod.[27] Oil con en was de e mined by he Soxhle me hod using hexane.[28] The oil con en has been exp essed in wo ways: he fi s one e e ed o he we weigh o he pas e (CAH) and he o he e e ed o he weigh o he d y pas e (CAS). Eu . J. Lipid Sci. Technol. 2021,123, 2000375 © 2021 Wiley-VCH GmbH 2000375 (2 o 11) www.ad ancedsciencenews.com www.ejls .com 2.4. Oil Samples Th ee ep esen a i e oli e samples, each weighing a minimum o 1 kg, we e p ocessed and he co esponding EVOOs we e ob- ained using an Abenco milling sys em (Abengoa S. A., Se illa, Spain). The oli e ui s we e milled using a s ainless-s eel ham- me mill equipped wi h a 5 mm sie e ha was ope a ed a 3000 pm. The esul ing oli e pas e was immedia ely kneaded in a mixe a 50 pm o 30 °C, wi h ho wa e added a 20 min. Cen i uga ion o he kneaded oli e pas e was pe o med in a bas- ke cen i uge a 3500 pm o 1 min. A e cen i uga ion, he oil was decan ed and s o ed in ambe glass bo les a 4 °C in da kness and wi hou headspace un il analysis. 2.5. S abili y The oxida i e s abili y was de e mined acco ding o he Ranci- ma me hod [29] using a Rancima appa a us (Me ohm, He isau, Swi ze land). 2.6. Analy ical Indices F ee acidi y, pe oxide alue, and UV spec opho ome ic indices (K232, K270) we e e alua ed acco ding o he official me hods desc ibed in Regula ion EEC 2568/91 and subsequen amend- men s o he Commission o he Eu opean Union.[4] All pa am- e e s we e de e mined in iplica e o each sample. 2.7. Fa y Acid Analysis The me hodology used was a modifica ion o he p o isions o Commission Regula ion (EEC) 2568/91.[4] The supe na an so- lu ion con aining he a y acid me hyl es e s was injec ed in o an Agilen 6890 se ies gas ch oma og aph equipped wi h an au- oma ic sample (Agilen 7683), spli -spli less injec o , flame ion- iza ion de ec o (FID) and HP Chems a ion (Agilen Technology) p og am. The a y acids we e quan ified in pe cen ages. Thei iden ifica ions we e made by compa ing e en ion imes wi h a s anda d solu ion o he me hyl es e s o pu e a y acids. 2.8. Polyphenols and Tocophe ols Analysis The con en o ocophe ols was analyzed acco ding o he IUPAC 2432 me hod[30] by high-pe o mance liquid ch oma og aphy (HPLC) using a Pe kin-Elme liquid ch oma og aph equipped wi h an isoc a ic LC200 pump and an LC295 UV–Vis de ec o . The esul s we e exp essed as mg o ocophe ol pe kg o oil.[31] The de e mina ion o o al polyphenols was pe o med ol- lowing he me hod adop ed by he In e na ional Oleic Coun- cil o he chemical analysis o he biophenols o oli e oil. This echnique is based on he ex ac ion o he mino pola compo- nen s o a phenolic na u e di ec ly om oli e oil by means o a me hanolic solu ion and i s subsequen quan ifica ion by HPLC wi h a UV de ec o a 280 nm, using sy ingic acid as he in e - nal s anda d. The o al con en o phenolic componen s (na u al o oxidized de i a i es o oleu opein and ligus oside, in lignans, in fla onoids and in phenolic acids) is exp essed in mg kg-1 o y osol. 2.9. Senso y Analysis The o ganolep ic assessmen s o his s udy we e ca ied ou by he as ing panel o he Regula o y Council o he P o ec ed Designa ion o O igin “P iego de Có doba,” in acco dance wi h he IOC s anda ds. The as ing panel was o med by a g oup o as e s, be ween 8 and 12, p e iously selec ed and ained acco ding o he echniques p e-es ablished by he IOC/ .20/ Doc n° 15 / Re .2 Sep embe 2007, and acco ding o he Eu o- pean Regula ion CE 640/2008.[32] The senso y p ofile o each EVOO sample was exp essed as he median alue o each desc ip o . The as e s e alua ed di ec ly o e onasally a oma ic ol ac o y sensa ions, such as oli e ui y, g een lea / esh-cu g ass, apple, almond, a ichoke, and o he posi i e a ibu es, such as gus a o y sensa ions (bi e ness and swee ness) and ac- ile/kinaes he ic sensa ions (pungency). The as e s had o a e he in ensi y o he diffe en desc ip o s on a con inuous scale. 2.10. Vola ile Analysis The de e mina ion o ola ile compounds was ca ied ou by gas ch oma og aphy coupled o mass spec ome y (GC-MS), ollow- ing and adap ing he me hod p oposed by Callejon e al.[33] 2go oli e oil was placed in headspace ials o 20 mL capaci y om Ge - s el (Müllheim and de Ruh , Ge many), and a e inse ing he wis e in a glass inse , i was igh ly closed and hea ed in a he - mos a ic ba h a 60 °C o 1 h. GC-MS analysis was pe o med by a 6890 Agilen GC sys em coupled wi h a quad upole mass spec- ome e Agilen 5975ine and equipped wi h a Ge s el The mo Deso p ion Sys em (TDS2) and a c yo- ocusing CIS-4PTV injec- o (Ge s el). The quad upole, sou ce, and ans e line empe a- u es we e main ained a 150 °C, 230 °C, and 280 °C, espec i ely. The elec on ioniza ion mass spec a in he ull-scan mode we e eco ded a 70 eV wi h he elec on ene gy in he ange o 29 o 300 m/z. The iden ifica ion o ola ile compounds was based on mass spec um ma ching o he NIST 98 lib a y and confi med by he a ailable e en ion index o s anda ds (RIs). When s anda ds we e no a ailable, he compounds we e iden ified by compu e ma ching o he e e ence mass spec a om he NIST lib a y and by he compa ison o hei RIs wi h he RIs ob ained wi h s anda ds ha ha e been epo ed in he li e a u e. The emain- ing compounds we e en a i ely iden ified by compu e ma ch- ing o he e e ence mass spec a om he NIST lib a y and/o h ough compa isons o hei RIs wi h hose o online da abases (Fla o ne ; Phe obase) and he li e a u e. RIs we e calcula ed by using he e en ion imes o n-alkanes ob ained unde iden ical analy ical condi ions, acco ding o he li e a u e.[34] The samples we e analyzed by iplica e. 2.11. S a is ics All s a is ical analyses and da a ea men s we e ca ied ou by using he PLS_Toolbox 7.9.5 wo king unde MATLAB en i on- men . The significance o he diffe ences be ween he means a he 5% le el was de e mined by pe o ming an analysis o a iance (ANOVA) o a ac o and he Tukey “PosHoc” es . Eu . J. Lipid Sci. Technol. 2021,123, 2000375 © 2021 Wiley-VCH GmbH 2000375 (3 o 11) www.ad ancedsciencenews.com www.ejls .com Figu e 1. Spide cha s ob ained by he senso y analysis o he a) oli e oils, and b) sco es and c) loadings plo s ob ained by a PCA ca ied ou wi h he a eas o he ola ile compounds iden ified in all he samples acco ding o he a ie y (Picual-P o Hojiblanca-H), cul i a ion modali y (con en ional wi h i iga ion a ming (CI), o ganic wi h i iga ion a ming (OI), and con en ional wi hou i iga ion o d yland a ming (CD)) and ipening s age (I, II, III, IV). P incipal componen analysis (PCA) was applied o he diffe en pa ame e s s udied as well as o he global pa ame e s o analy- ses, in o de o s udy he influence on he oli e oil acco ding o he diffe en cul i a ion modali ies (d yland, con en ional i iga- ion, and o ganic i iga ion), he diffe en momen s o ui ha - es ing and he diffe en a ie ies. Au oscaling and g oup-scaling we e applied o he indi idual and he global da a ma ices, e- spec i ely. 3. Resul s and Discussion 3.1. Chemosenso y E alua ion The senso y analysis and he e o e, he ola ile p ofile o oli e oils has an impo an ole o he quali y and p ope ies o he comme cialized p oduc . Fo his eason, a chemosenso y e al- ua ion was pe o med in de ail. Figu e 1 shows, on he le side, he spide cha s ob ained by he senso y analysis o he oli e oils acco ding o he a ie y, cul i a ion modali y, and ipening s age (Figu e 1a), and on he igh side, he sco es and loadings plo ob ained by a PCA ca ied ou wi h he a eas o he ola ile com- pounds iden ified in he samples (Figu e 1b,c). Conce ning he senso y p ofiles o he oli e oil, i can be seen how hey a y as ipening p og ess (Figu e 1a), acco ding o ha epo ed by se e al au ho s.[10,35] Thus, o bo h a ie ies and modali ies, he swee a ibu e showed an inc easing end h ough ipening, acco ding o he li e a u e[36,37] and eaching, in gene al, he maximum alues in s age III. Bi e and ui y a - ibu es also eached maximum alues in s age III o Picual oils, acco ding o Jiménez e al.[37] As o he au ho s epo ed,[36] he g ass, apple, and almond a ibu e ended o dec ease wi h ipen- ing in bo h a ie ies, eaching he lowes sco es in he las ipen- ing s ages. The emaining a ibu es did no show a clea end wi h ipening, a ying acco ding o he a ie y o he cul i a ion modali y. Rega ding he a ie y, he e we e also some diffe ences in hei senso y p ofile. Thus, in gene al and conside ing mean alues, he Hojiblanca a ie y was mo e pungen , g assie , and swee e han he Picual, which eached g ea e alues o he bi e , ap- ple, almond, and lea a ibu es. The senso y diffe ences be ween hese a ie ies we e also obse ed by o he au ho s. [10,38] The ui y desc ip o showed simila alues in bo h a ie ies. Acco d- ing o Dabbou e al.,[38] he ui y a ibu e is a eflec ion o he aw ma e ial om which he oil is ob ained and i is undamen al in he VOO a oma, since he a oma o fla o o VOO is p oduced by he balance be ween g een and ui y no es. Wi h espec o he cul i a ion modali y, he e we e ewe diffe ences in gene al. Hence, he d yland modali y (CD) p e- sen ed lowe mean sco es o mos o he senso y a ibu es, s anding ou o a highe swee ness, in acco dance wi h o he au ho s.[38] In compa ison, he con en ional oils ob ained unde i iga ion (CI), eached highe alues in bo h a ie ies o he Eu . J. Lipid Sci. Technol. 2021,123, 2000375 © 2021 Wiley-VCH GmbH 2000375 (4 o 11) www.ad ancedsciencenews.com www.ejls .com ui y, pungen , bi e , g ass, and lea a ibu es, especially in s age III (Figu e 1a). These esul s a e also in acco dance wi h Rosa i e al.,[39] Jiménez e al.,[10] and Dabbou e al.[38] who obse ed an inc ease in hese a ibu es when inc easing he i iga ion le el. The o ganic oils (OI) showed, in gene al, highe alues o bi e ness and ui y a ibu es, mainly in he second ipening s age. This is due o he highe p esence o phenols and ola ile compounds syn hesized h oughou he lipoxygenase (LOX) pa hway, which a e esponsible o bi e and oli e ui y senso y a ibu es, espec i ely.[10,40] The highe p esence o hese a ibu es a o s he quali y o hese oils in compa ison o he oils ob ained by he o he modali ies.[41] The diffe ences be ween a ie ies and modali ies can be clea ly obse ed by looking a he ipening s age I. Hence, PCD oil is highligh ed by i s swee no es, PCI by i s pungen , he b, ui y, and apple no es, and POI oil by i s bi e ness (Figu e 1a). This was expec ed, since, as men ioned abo e, oils om o ganic a m- ing eached highe amoun s o o al phenols (Table S2, Suppo - ing In o ma ion), which has shown o be ela ed o he bi e a ibu e.[42] On he o he hand, Hojiblanca in his fi s s age is highligh ed by highe lea no es in HCI, mo e he b no es in HOI and swee and apple no es in HCD oils. Mo eo e , s age III was he one ha showed ewe diffe ences among he diffe en modali ies. In gene al, he be e senso y sco es we e ob ained o he ipening s ages III and IV excep o he o ganic oils whose be - e senso y p ofiles we e ob ained in s ages II and III. This di - e ence is in acco dance wi h he diffe ences discussed abo e e- ga ding he ipening o he o ganic ui s, which ma u ed ea lie han ui s o he o he modali ies. Table S3 in Suppo ing In o ma ion showed he mean o o- al peak a eas ob ained o he ola ile compounds de ec ed, hei s anda d de ia ion and he s a is ical esul s ob ained by ca ying ou an ANOVA ega ding he a ie y, ipening s age, and cul i a- ion modali y. In ou s udy, a o al o 85 ola ile compounds we e de e - mined o bo h a ie ies, all cul i a ion modali ies and ipen- ing s ages, 3 o which a e desc ibed and iden ified o he fi s ime in EVOOs (1-hyd oxy-2-p opanone, (E)-linalool oxide, and 2-ace yl u an). Wi hin he 85 ola ile compounds de e mined, 15 compounds we e alcohols, 14 ke ones, 10 u ans, 8 aldehydes, 8 e penes, 8 hyd oca bons, 7 es e s, 7 acids, 3 lac ones, and 5 compounds classified as “o he .” In gene al e ms, ega ding he alues o o al peak a eas ob- ained o he o al ola ile compounds in bo h a ie ies (Table S3, Suppo ing In o ma ion), Hojiblanca seemed o ha e a signifi- can ly highe alue o ola ile compounds han Picual samples, clea ly obse able in he alcohols, aldehydes, and es e s ami- lies. Howe e , Hojiblanca samples showed lowe alues o acids, u ans, and hyd oca bons han Picual. Rega ding he modali- ies, EVOOs om con en ional cul i a ion (i iga ion-CI- and d yland-CD) had highe o al a eas o he o al ola ile composi- ion han he o ganic EVOOs, excep in s age I o bo h a ie ies, whe e he opposi e was obse ed. Mo eo e , in con en ional EVOOS o bo h a ie ies, he o al a eas o mos o he chemi- cal amilies ollowed he same end o inc easing wi h ipening, showing hei maximum in Picual IV and Hojiblanca III. These esul s ma ched he esul s ob ained by Jimenez e al.[10] How- e e , o ganic samples showed diffe ences be ween Picual and Hojiblanca a ie ies, since he o al a ea o ola ile compounds in Picual significan ly inc eased wi h ipening while i dec eased in Hojiblanca. Rega ding he i iga ion effec , al hough bo h con en ional modali ies showed an inc easing end wi h ipening, in gen- e al, i was obse ed ha hose EVOOs om bo h a ie ies un- de i iga ion cul i a ion (PCI and HCI) eached highe alues o o al ola ile compounds han hose unde d yland cul i a ion (PCD and HCD) (Table S3, Suppo ing In o ma ion). Acco ding o he li e a u e, he e is no clea end o effec wi h ega d o he change in o al ola ile compounds as a unc ion o he i iga ion egime.[38] As is well known, he majo ola ile compounds e- po ed in i gin oli e oils a e he C6 and C5 compounds de i ed om linoleic and linolenic acids h ough he lipoxygenase (LOX) pa hway, which akes place du ing he c ushing o he oli e ui and oli e pas e malaxa ion.[6,43] These compounds, e e ed o as LOX compounds, a e mainly aldehydes, alcohols, es e s, hyd o- ca bons, and ke ones, which a e esponsible o he cha ac e is- ic and posi i e g een senso y no es. This g oup o compounds, con a y o ha obse ed by Jiménez e al.[10] inc eased h ough he ui ipening p ocess in con en ional EVOOs (d yland and i iga ion), eaching he highes alues in s ages III and IV in hose om Hojiblanca and Picual a ie ies, espec i ely (Figu e S1, Suppo ing In o ma ion). Howe e , o ganic EVOOs did no p esen a clea end h ough ui ipening. Nin ali e al.[44] com- pa ed con en ional and o ganic oli e oils ac oss a 3 yea s udy and a nonconsis en end was obse ed because he ola ile compo- si ion depends on a a ie y o ac o s, including cul i a , clima e, soil ype, nu ien and wa e a ailabili y, du a ion, and condi ions o s o age and he diffe ences ela ed o he cul i a ion me hods may be difficul o be obse ed.[45] Mo eo e , ega ding he i i- ga ion effec on LOX compounds, no significan diffe ences we e obse ed be ween he i iga ed and d yland samples. As shown in Table S3 (Suppo ing In o ma ion), mos o he ola ile compounds showed significan diffe ences be ween o al a eas. Hence, a PCA was de eloped in o de o be e s udy he diffe ences be ween a ie ies, cul i a ion modali ies, and ipen- ing s ages conside ing he ola ile p ofile. The sco es and load- ings plo s ob ained by he second and hi d PCs (explaining 24.82% o a iance) we e epo ed in Figu e 1b,c (on he igh side o Figu e 1). Hence, i could be clea ly obse ed ha he ola ile composi ion diffe en ia ed he samples mainly h ough hei a ie y (explained by PC2). This esul was expec ed since a ious au ho s ha e epo ed he gene ic effec ela ed o cul i- a as one o he mos impo an aspec s o he ola ile composi- ion o oli e oils.[46] Mo eo e , diffe ences h ough he ipeness s age (explained by PC3) could be obse ed. Thus, he fi s ipen- ing s ages I and II we e placed on he uppe side o he sco es plo (posi i e alues o PC3) and he la es s ages on he lowe side (nega i e alues o PC3). In addi ion, in bo h a ie ies, bu mainly in Picual, i could be obse ed ha in he fi s s ages he e we e no many diffe ences ega ding he cul i a ion modali ies, and he samples appea ed mo e g ouped. Howe e , as ui ipen- ing inc eased, a g ea e diffe ence in he ola ile composi ion was gene a ed, especially in he case o o ganic a ming wi h i iga- ion (OI). Wi hin he amily g oups wi h he highes numbe o com- pounds, he alcohol amily was he majo i y g oup ollowed by ke ones and aldehydes. Alcohols ha e been desc ibed as he class Eu . J. Lipid Sci. Technol. 2021,123, 2000375 © 2021 Wiley-VCH GmbH 2000375 (5 o 11) www.ad ancedsciencenews.com www.ejls .com Figu e 2. Sco es and loadings plo s o he PCA model ob ained wi h he o al o pa ame e s (physicochemical (PC), ag onomical (AGR), a y acids (FA), polyphenols and ocophe ols (P&T), ola ile compounds (VC) and senso y sco es (SENS) and samples acco ding o he a ie y (Picual (P) o Hojiblanca (H)), cul i a ion modali y (con en ional wi h i iga ion a ming (CI), o ganic wi h i iga ion a ming (OI) and con en ional wi hou i iga ion o d yland a ming (CD)) and ipening s age (I, II, III, IV)) analyzed. wi h he la ges a ie y o ola ile compounds in oli e oils in qual- i a i e e ms.[38] In ou s udy, mos o hem p o ed o be mo e ela ed o he Hojiblanca a ie y (Figu e 2c), being 1-pen en-3- ol, 1-pen anol, 1-hexanol, (E)-3-hexen-1-ol, (Z)-3-hexen-1-ol, and (E)-2-hexen-1-ol mo e ela ed o he fi s ui ipening s ages and e hanol, me hanol, (E)-2-pen enol, (Z)-2-pen en-1-ol, (E)-2- decenol, and 1-undecanol o he las ma u i y s ages. Rega ding he cul i a ion modali ies, he significan diffe - ences in alcohols we e obse ed in s ages III and IV o Hoji- blanca oils. Thus, in Hojiblanca a ie y, in s age III he o ganic samples p esen ed significan lowe a eas o alcohols han he con en ional ones while in s age IV alcohols highligh ed in he o ganic oils, acco ding o Jiménez e al.[10] Compa ing he con- en ional modali ies o he Hojiblanca a ie y, a significan effec o he i iga ion egime was obse ed in s age III, eaching highe alues o alcohols in he ain ed samples. This esul was in con- co dance wi h Sanchez-Rod íguez e al.[47] who ound an inc ease in alcohol concen a ion in all s essed oli e oils due o a defici i iga ion ea men . In Picual, significan diffe ences in he al- cohol o al a eas among he h ee modali ies we e no obse ed a any ipening s age. Wi hin he ke ones, he second g oup wi h he highes num- be o compounds, significan diffe ences among he cul i a ion modali ies we e no obse ed a any s age. 2-Pen anone, which has been ecognized as one o he main compounds esponsi- ble o ui y no es,[47] showed diffe en alues be ween bo h a- ie ies, mainly in he fi s s ages, eaching he Hojiblanca a ie y highe peak a eas. Hence, acco ding o ha , i could be ela ed o he ema ked ui y no es o Hojiblanca oils in he fi s s ages (Figu e 1a). Aldehydes seemed o be mo e p esen in he las s ages o bo h a ie ies (Figu e 2b,c), being consis en wi h he li e a u e.[10] Thus, he u anic aldehydes such as 3- u aldehyde, u u al, and me hyl u u al we e mo e ela ed o Picual oils o he las s ages, and hexanal, (Z)-3-hexenal, (E)-2-hexenal, benzaldehyde, (E)-2- decenal o Hojiblanca oils o he las s ages. O he au ho s ha e also demons a ed ha (Z)-2-hexenal, conno ed by a ui y no e, and (Z)-3-hexenal, wi h a g een odo con ibu e o he disc imi- na ion a he ipening s age.[48] Mo eo e ,acco ding oAnge osa e al.,[49] hexanal, (Z)-3-hexenal, and (E)-2-hexenal ha e p o ed o be one o he mos impo an con ibu o s o high quali y o EVOOs. Howe e , i should be also highligh ed ha al hough hex- anal has been ela ed o g een apple and g ass senso y desc ip- o s, allowing he diffe en ia ion o EVOOs om he es o he ca ego ies, his compound has also been ela ed o a mus iness- humidi y, us y, winey- inega y, o ancid de ec , depending on i s concen a ion.[50] Mo eo e , (E)-2-hexenal, ela ed o g een no es,[38] can be highligh ed o i s high a eas in bo h a ie ies in he las ipening s ages, in acco dance wi h o he au ho s.[12] In addi ion, ou esul s we e also consis en wi h he li e a u e, showing ha , especially in he Picual a ie y, i iga ed samples ex- hibi ed highe a eas o (E)-2-hexenal and hexanal han he ain ed ones.[12,38] Finally, he emaining chemical classes, wi h lowe numbe o compounds han he o he g oups, could be conside ed as mino compounds, as o he au ho s ha e indica ed.[38] Wi hin hem, es- e s we e mo e p esen a he las s ages o he Hojiblanca a i- e y such as 1-me hoxyhexane, hexyl ace a e, me hyl dihyd ojas- mona e, 1-me hoxy-(Z)-3-hexene and (Z)-3-hexenyl ace a e. This las compound, pa icula ly co ela ed o senso y a ibu es like g een lea es, could be highligh ed due o i ha ing p o ed o be ele an in he diffe en ia ion be ween non-de ec i e (EVOO) and de ec i e (non-EVOO) oli e oil samples, being p esen in mo e concen a ion in EVOO samples.[50] The eason o i s in e es elies on i s odo h eshold. Thus, (Z)-3-hexenyl ace a e has a much lowe odo h eshold concen a ion (be ween 1 and 2 𝜇g Eu . J. Lipid Sci. Technol. 2021,123, 2000375 © 2021 Wiley-VCH GmbH 2000375 (6 o 11) www.ad ancedsciencenews.com www.ejls .com L-1) han i s co esponding alcohol, (Z)-3-hexen-1-ol (50–70 𝜇g L-1),[51] which means ha in low concen a ions i could p o ide impo an g een no es o he oli e oil, use ul o he senso y di - e en ia ion o EVOOs. In he Picual a ie y, some u ans and acids (2-me hyl u ane, 2-ace yl u an, 5-hyd oxyme hyl u u al, and bu y ic acid, pen- anoic acid and 2-e hylhexanoic acid) highligh ed in he fi s s ages, while o he s such as ace ic acid, p opanoic acid, and hexanoic acid, which a e ela ed o senso y de ec s,[52] we e mo e p esen in he las s ages. Ano he ola ile compound ha has been ela ed o senso y de ec s is e hyl ace a e (winey, inega y).[50,52] Al hough his compound has p o ed o be ele- an in he diffe en ia ion o EVOO and non-EVOOs, due o i showing mo e concen a ion in LOO samples h ough a GC-IMS analysis,[50] in he p esen s udy i showed low a eas in gene al e ms o all he samples, so i migh no influence he senso y p ofile. In summa y, we can say ha he a ie y had a majo influ- ence on he ola ile and senso y p ofiles in compa ison wi h he cul i a ion modali ies, he diffe ences o a ie ies being mainly ela ed o a ew C5 and C6 alcohols and aldehydes, and ui y, bi e , and pungen senso y a ibu es as has occu ed in o he s udies.[38] 3.2. O e all E alua ion o Quali y The ag onomical and physicochemical pa ame e s and he con- en o polyphenols, ocophe ols, and a y acids a e summa ized in Tables S1, S2, and S4 (Suppo ing In o ma ion). Acco ding o hese esul s, he fi s s ep was o check ha all he samples p e- sen ed ce ain alues below he limi s se led o EVOO samples by he In e na ional Oli e Council s anda ds (IOC, 2013).[4] Thus, as all hese pa ame e s ha e al eady been s udied by o he au ho s in hese wo a ie ies, as well as in he diffe en cul- i a ion modali ies and diffe en ipening s ages, we conside ed s udying hem simul aneously in o de o make an o e all e al- ua ion o how hese pa ame e s influence he diffe en a ie ies, cul i a ion modali ies and ipening s ages and he ela ionships be ween hem. Mo eo e , i should be emembe ed ha his was he fi s ime ha he influence o i iga ion and p oduc ion sys- em (o ganic and con en ional) on he quali y o EVOOs om he Hojiblanca and Picual a ie ies and diffe en ui ipening deg ees, ob ained unde simila condi ions, was also s udied si- mul aneously. Fo his pu pose, a PCA model was applied o he o al numbe o conside ed pa ame e s and samples analyzed in his s udy. The co esponding sco es and loadings plo a e shown in Figu e 2. The sco es plo shows, as expec ed, ha he a ie y was he mos influen ial a iable wi h espec o he i iga ion o cul i- a ion modali y, being he fi s p incipal componen (PC1 wi h a 28.20% o o al a iance), esponsible o his diffe en ia ion. Thus, samples we e g ouped acco ding o he a ie y, Picual sam- ples being placed on he posi i e side and Hojiblanca samples on he nega i e side o PC1. I could also be obse ed ha he second p incipal componen , PC2, which explained he 20.88% o he o al a iance, diffe en- ia ed he samples acco ding o he ipening s age, being he la - es s ages on he posi i e PC2 side, and he ea lies ones on he nega i e side, o all he modali ies and bo h a ie ies. Mo eo e , a sligh diffe ence could also be app ecia ed be ween cul i a ion modali ies bu wi hin each a ie y. Conce ning he loadings, a ie al diffe ences we e mainly e- la ed o he highe p esence o bi e and lea senso y a ibu es, bu also o a highe p esence o oleic, palmi ic, and palmi oleic a y acids, highe alues o acidi y, s abili y, and pe oxide index in Picual oils oge he wi h he highe alues o CAH and CAS o Picual ui s, whe eas Hojiblanca oli e oil showed a highe p es- ence o apple, pungen and swee no es, a highe con en in o- cophe ols, linoleic, linolenic, and o he a y acids, and a highe esis ance and humidi y alues in i s ui s. This diffe en ia ion also occu ed when he diffe en g oups o pa ame e s we e s udied sepa a ely, being consis en wi h he esul s ob ained by o he au ho s ha app ecia ed a highe diffe - ence be ween a ie ies han be ween o he condi ions.[10,14,36–38] Hence, when he o al polyphenol and ocophe ol con en was s udied (Table S2, Suppo ing In o ma ion), i did no dis inguish EVOOs in acco dance wi h he cul i a ion modali ies, bu showed he main diffe ences be ween a ie ies (i.e., Hojiblanca showed mo e con en o o al ocophe ols and Picual mo e concen a ion o o al polyphenols), in ag eemen wi h Jiménez e al.[25] Some hing simila occu ed when he o al amoun o a y acids ob ained was s udied by sepa a ely (Table S4, Suppo ing In o ma ion). In gene al, bo h a ie ies clea ly showed he p e- dominance o monounsa u a ed a y acids (be ween 55.0% and 83.0% o he o al he eo ), oleic acid being he mos abundan a y acid in all he samples (Table S4, Suppo ing In o ma- ion). Howe e , he a ie y seems o be he mos impo an and de e mining ac o in he diffe ences in a y acids. Thus, he o al con en o a y acids was in gene al highe in he Hojiblanca samples, excep o palmi ic and palmi oleic acids whose con en s we e highe in he Picual a ie y (Table S4, Suppo ing In o ma ion), as was obse ed in o he s udies.[25,36] By he con a y, he e was a diffe ence in he oleic acid con en : he Picual a ie y had, in gene al, 5% mo e oleic acid han he Hojiblanca a ie y, excep in o ganic cul i a ion (POI) o which bo h a ie ies showed simila con en wi h sligh ly lowe alues han he o he wo modali ies.[36] Mo eo e , he palmi ic/linoleic a io, ha is ela ed o he oil balance, and oleic/linoleic a io, ela ed o s abili y,[53] we e g ea e o he Picual a ie y han o Hojiblanca in all modali ies and ipening s ages (Table S4, Suppo ing In o ma ion), in ag eemen Jiménez e al.[25,36] The e o e, i can be in e p e ed ha he Picual a ie y gene a es mo e balanced and s able oils han he Hojiblanca a ie y. How- e e , he linoleic/linolenic ela ion, ha is ela ed o he senso y a ibu es o oli e oils[54] (since hey gene a e he mos impo an ola ile compounds wi h “as ingen and g een” no es h ough he lipoxygenase pa hway), was g ea e in he Hojiblanca a ie y, in acco dance wi h o he au ho s.[35] A e s udying all he samples oge he , each a ie y was s ud- ied sepa a ely by he de elopmen o wo news PCAs (Figu e 3) in o de o see he diffe ences mo e clea ly be ween modali ies o ipening. The sco e plo in his case shows he o ganic a ming (OI) clea ly diffe en ia ed om he o he wo con en ional modal- i ies (CD and CI) by means o PC2 in bo h Picual and Hojiblanca PCA models. By s udying he loadings o bo h PCAs, he o ganic samples (placed on he nega i e o posi i e side o PC2 in he Picual and Eu . J. Lipid Sci. Technol. 2021,123, 2000375 © 2021 Wiley-VCH GmbH 2000375 (7 o 11) www.ad ancedsciencenews.com www.ejls .com Figu e 3. Sco es and loadings plo s o he PCA analysis de eloped wi h he o al o pa ame e s analyzed o he o al se o samples o he wo a ie ies a) “Picual”(P)andb)“Hojiblanca” (H), cul i a ed in h ee modali ies: con en ional wi h i iga ion a ming (CI), o ganic wi h i iga ion a ming (OI) and con en ional wi hou i iga ion o d yland a ming (CD); and ha es ed a 4 diffe en s ages o ma u i y (I, II, III, IV). Hojiblanca PCA, espec i ely) we e cha ac e ized in gene al by a highe weigh o 100 ui s, acidi y, CAS, K270, and K232 alues and ipening index; highe ui y, almond and bi e no es; and a highe concen a ion o s ea ic, palmi oleic and linoleic acids han he con en ional modali ies (d yland and i iga ion). Acco ding o hese esul s, a highe ela ion could be obse ed be ween he ag onomical and physicochemical pa ame e s abo e men ioned wi h he cul i a ion modali y han wi h he a ie y (Figu e 2). The g ea e weigh o he ui s wi h i iga ion compa ed o he d yland modali y migh be due o he addi ional wa e supply ha hey had,[21] usually highe in he o ganic modali y. In consequence, he o ganic cul i a ion showed sig- nifican ly lowe alues o shedding esis ance o i s ui s han he o he wo con en ional modali ies (Table S1, Suppo ing In o ma ion). The highe alues o ee acidi y in o ganic oli e oils we e also obse ed by o he au ho s,[22] and i was ela ed o a ce ain in es a ion o ungal diseases in he oli es esul ing om he absence o pes icides in his ag icul u al modali y. In con as , he pe oxide index showed in gene al highe alues in he con en ional modali ies, eaching highe alues o d yland a ming (CD) han i iga ion (CI), while he o ganic modali y showed he lowes alues (Table S1, Suppo ing In o ma ion), as occu ed in o he wo ks.[55] Wi h ega d o s abili y, he cul i a- ion modali y, and he e o e i iga ion, had an opposi e influence on bo h a ie ies (Table S1, Suppo ing In o ma ion). Thus, POI oils showed he sho es s abili y imes o all s ages, while HOI oils had he g ea es s abili y agains oxida ion in mos ipening s ages (Table S1, Suppo ing In o ma ion), in acco dance wi h he esul s epo ed by Jiménez e al.[14] Mo eo e , o ganic modali y also showed he highes con en o o al polyphenols in bo h a ie ies (mainly in POI and s age II) as was obse ed by o he au ho s,[2,10,23,39] ollowed by he con en ional i iga ion,[21,56,57] and eaching he lowes alues in he d yland modali y (Table S2, Suppo ing In o ma ion). In ac , i has been shown ha he ac i i y o he enzyme L-phenyllanine ammonia lyase (PAL), esponsible o he biosyn hesis o Eu . J. Lipid Sci. Technol. 2021,123, 2000375 © 2021 Wiley-VCH GmbH 2000375 (8 o 11) www.ad ancedsciencenews.com www.ejls .com phenolic compounds, is inc eased wi h high le el o i iga ion.[58–60] Howe e , o he au ho s ha e epo ed di - e en o e en opposi e esul s, obse ing no effec , a iable effec s o e yea s, o e en a dependency o he effec on he g owing a ea.[61] Hence, his b ings o ligh ha i iga ion does no always p oduce he same effec in he phenolic con en o oli e oils. Some hing simila occu ed wi h he ocophe ol con- en , no ollowing a clea end wi h ui ipening, o cul i a ion modali ies, as o he au ho s epo ed.[40] Rega ding a y acids, he con en ional modali ies we e again mo e simila , al hough d yland condi ions showed highe alues han i iga ion, and bo h diffe ing om o ganic oils. The la e showed lowe amoun s o oleic acid (as men ioned abo e) and highe con en s o linoleic acid (being s a is ically significan in he Hojiblanca a ie y), palmi ic and palmi oleic acids (mainly in he Picual a ie y) and s ea ic acid han he o he modali ies (CI and CD). These diffe ences we e in acco dance wi h he esul s o Jimenez e al.,[36] bu hey con as ed wi h he esul s ob ained by o he au ho s ha did no obse e diffe ences in a y acid composi ion,[62] o e en ha a y acid composi ion in he oils om he i iga ion ea men s a ied wi h he seasons.[63] Mo e- o e , among he h ee modali ies s udied, he highes alues o linoleic/linolenic a io (Table S4, Suppo ing In o ma ion) we e eached in hose wi h ex a wa e supply, and among hem, being s a is ically significan in he Picual o ganic cul i a ion (POI), and hese esul s we e in acco dance wi h hose ob ained by se e al au ho s.[23,36,63] Finally, in bo h global PCA model (Figu e 2) and he indi idual PCA model o each a iable (Figu e 3), he ipening effec was obse ed. Thus, he ea lies ipening s ages showed highe ela- ion wi h g ass, almond, bi e , and pungen senso y a ibu es, o al polyphenols, s abili y, and oleic acid; while he la es ipen- ing s ages we e mo e ela ed o swee , ui y, and apple senso y a - ibu es, ocophe ols, ipening index, weigh o 100 ui s, CAH, CAS, and humidi y, as well as acidi y and linoleic and linolenic acids. The e o e, hese la es s ages showed lowe alues o shed- ding esis ance, pe oxide index, and s abili y. The CAS, CAH, and humidi y pa ame e s could be ele an due o i ha ing been demons a ed ha humidi y influences he con en o phenolic compounds. Thus, o he wo ks showed ha when humidi y inc eased significan ly, phenolic compounds dec eased.[40] Rega ding he s abili y o oli e oil, ou esul s showed ha he oils ob ained om oli es a mo e ad anced s ages o ipening showed lowe s abili y (Table S1, Suppo ing In o - ma ion). This end was also obse ed in o he s udies and could be explained by a educ ion in wa e con en wi h ipening.[14,40,64] The esis ance o oxida i e de e io a ion is gene ally a ibu ed o he lipid composi ion and he an ioxidan con en (i.e., phenolic compounds).[65] Mo eo e , g ea e diffe ences could be seen be ween modal- i ies han be ween a ie ies ega ding he ipeness deg ee (Fig- u e 2), wi h some o he ag onomical and physicochemical pa- ame e s eaching he highes alues in all he ipeness s ages o he o ganic samples (Table S1, Suppo ing In o ma ion), mean- ing ha he o ganic samples migh an icipa e a s age o ma u i y. A common end was also obse ed o bo h a ie ies and cul i- a ion modali ies, going om he nega i e side o PC1 and PC2, s ages I and II, o he posi i e side o PC1 and PC2, wi h he high- es ipening s ages (Figu e 3a,b). Mo eo e , al hough he concen a ion o polyphenols among he ipening s ages was diffe en acco ding o he a ie y (i.e., Picual showed an inc ease om s age I o II, and hen i de- c eased while Hojiblanca showed a dec ease in o al polyphenols om s age I o IV) (Table S2, Suppo ing In o ma ion), i ol- lows he same end al eady s udied in he li e a u e: a p og es- si ely inc ease du ing ipening, un il i eaches a maximum a he “spo ed” and “pu ple” pigmen a ion s age, a e which i de- c eases, ollowing a simila end o s abili y and humidi y.[40] Las ly, an influence o ipening in he a y acid was no clea ly obse ed (Table S4, Suppo ing In o ma ion), emained p ac i- cally unchanged as ipening p og essed, as was also obse ed by Jiménez e al.[25] Howe e , i could be obse ed ha he a ios palmi ic/linoleic and oleic/linoleic dec eased h oughou ma u- a ion in all modali ies and a ie ies, in acco dance wi h Jiménez e al.[36] and o he au ho s[19] who epo ed ha monounsa u- a ed/polyunsa u a ed a ios we e no influenced by he i iga- ion le el. On he con a y, he linoleic/linolenic a io inc eased in bo h a ie ies and cul i a ion modali ies as ma u a ion inc eases, eaching he maximum alues in s age IV, and also showed diffe - ences be ween cul i a ion modali ies (mo e in o ganic samples). 4. Conclusions In his wo k, he influence o diffe en aspec s, such as he a- ie y, ui ipening deg ee, he i iga ion and he ype o cul i- a ion (con en ional and o ganic), on he quali y o he EVOO ob ained in simila condi ions (geog aphical a ea, clima e … e c) ha e been s udied simul aneously, aking in o conside a ion he ag onomical and physicochemical pa ame e s, he polyphenols, ocophe ols, and a y acid composi ion and he ola ile and sen- so y p ofiles. In he li e a u e, he e a e simila s udies, bu hey do no conside all hese a iables simul aneously, no de e mine all hese physicochemical pa ame e s, and in mos cases, oli e samples we e no ob ained unde simila condi ions. In addi ion, among he 85 ola ile compounds de e mined in he samples, 1-hyd oxy-2-p opanone, (E)-linalool oxide and 2- ace yl u an ha e been desc ibed o he fi s ime in oli e oil. The esul s ob ained showed ha oli e oils diffe ed fi s acco d- ing o hei a ie y, secondly o he s age o ipeness o he ui s ( he mo e he g ea e he ipening deg ee o he ui ) and finally, wi hin each a ie y, acco ding o he cul i a ion modali y. Thus, o ganic i iga ion diffe ed ma kedly om con en ional modali- ies (wi h and wi hou i iga ion), wi h significan diffe ences be- ing obse ed in he acidi y, s abili y, ocophe ol and polyphenol con en s, a y acid composi ion and senso y a ibu es. The a- ie y and he ipening deg ee o he oli es had an effec on all he pa ame e s s udied, showing a g ea e effec on he ola ile composi ion, and, he e o e, in hei o ganolep ic cha ac e is ics and quali y o EVOOs, han he cul i a ion me hod (o ganic o con en ional). I iga ion was he a iable ha p o ed o ha e he leas influ- ence on he chemical composi ion o he oli e oils. Howe e , in bo h a ie ies, i iga ion inc eased yields. F om a senso ial poin o iew, he senso y a ibu es o g ass, apple, bi e and pungen , made i possible o diffe en ia e he ea lies ipening s a es o he ui s a he ime o ha es ing and we e consis en wi h he amoun s ob ained in o al polyphenols, s abili y, as well as quali y pa ame e s. Eu . J. Lipid Sci. Technol. 2021,123, 2000375 © 2021 Wiley-VCH GmbH 2000375 (9 o 11)