Development of a fermented quinoa-based beverage

 www.foodscience-nutrition.com Food Sci Nutr. 2017;5:602–608.602 |  © 2016 The Authors. Food Science & Nutrition published by Wiley Periodicals, Inc. Received:13July2016 | Revised:15September2016 | Accepted:25September2016 DOI: 10.1002/fsn3.436 Abstract QuinoaisacropthatoriginatedfromtheAndes.Ithashighnutritionalvalue,outstandingagro-ecologicaladaptability,andlowwaterrequirements.Quinoaisanexcellent cropalternativetohelpovercomefoodshortages,anditcanalsohavearoleinthe preventionofdevelopedworldlifestylediseases,suchastype-2diabetes,cardiovasculardiseases,osteoporosis,inflammatoryandautoimmunediseases,etc.Inorderto expandthetraditionalusesofquinoaandtoprovidenew,healthierandmorenutritiousfoodproducts,afermentedquinoa-basedbeveragewasdeveloped.Twoquinoa varieties(RosadadeHuancayoandPasankalla)werestudied.Thefermentationprocess,viscosity,acidity,andmetabolicactivityduringthepreparationandstorageof thedrinkweremonitored,aswellasthepreliminaryorganolepticacceptabilityofthe product.Thedrinkhadviableandstablemicrobiotaduringthestoragetimeandthe fermentationprovedtobemostlyhomolactic.Bothquinoavarietiesweresuitableas baseforfermentedproducts;Pasankalla,however,hastheadvantageduetohigher proteincontent,lowersaponinconcentration,andlowerlossofviscosityduringthe fermentationprocess.Theseresultssuggestthatthedifferencesbetweenquinoavarietiesmayhavesubstantialeffectsonfoodprocessesandonthepropertiesoffinal products.Thisisafactorthatshouldbetakenintoaccountwhenplanningnovelproductsbasedonthisgrain. KEYWORDS beverage,fermentation,quinoa,starter,varieties 1FoodBiotechnology,InstituteofPublic HealthandClinicalNutrition,Universityof EasternFinland,Kuopio,Finland 2DepartmentofFoodTechnology,Faculty ofFoodEngineering,LaMolinaAgrarian University,Lima,Peru 3CEMIS-Oulu,UniversityofOulu,Kajaani, Finland 4GreenTechnology,LUKENaturalResources InstituteFinland,Jokioinen,Finland Correspondence CarmePlumed-Ferrer,FoodBiotechnology, InstituteofPublicHealthandClinical Nutrition,UniversityofEasternFinland, Kuopio,Finland. Email:[email protected] Funding information MinistryforForeignAffairsofFinland ORIGINAL RESEARCH Development of a fermented quinoabased beverage Fanny Emma Ludena Urquizo1,2 | Silvia Melissa García Torres1,2 | Tiina Tolonen3 | Mari Jaakkola3 | Maria Grazzia Pena-Niebuhr1,2 | Atte von Wright1 | Ritva Repo-Carrasco-Valencia2 | Hannu Korhonen4 | Carme Plumed-Ferrer1 1 | INTRODUCTION Quinoa(Chenopodium quinoaWilld.)isanancientgraincropthatoriginatedfromtheAndeanregionofSouthAmerica.Quinoabelongsto theChenopodiaceaefamilyandincludesaround250speciesand3,000 varieties conserved in germplasm banks (Vega-Galvez etal., 2010). Quinoahas anextremeagro-ecologicaladaptability: Itcanbe cultivatedbothincold,highlandclimates,andin subtropicalconditions; from sea level to above 4000m of altitude (Miranda etal., 2012; Repo-Carrasco, Espinoza, & Jacobsen, 2003). This property gives quinoaagoodpotentialtobeintroducedaroundtheworld.Jacobsen (2003)studiedthecultivationofdifferentvarietiesofquinoainNorth America,Africa,Asia,AustraliaandEurope,showingitsrealisticpotentialasanovelcropintheseregions. The extension of the global cultivation and uses of quinoa couldbe advisable, becausethegrainsarehighlynutritioushaving exceptional protein quality and awide range of vitamins and minerals.Quinoaproteinhasabalancedaminoacidcompositionbeing rich in essential amino acids such as lysine (5.1–6.4%) and methionine (0.4–3.1%). The total dietary fiber content of quinoa grains ThisisanopenaccessarticleunderthetermsoftheCreativeCommonsAttributionLicense,whichpermitsuse,distributionandreproductioninanymedium, providedtheoriginalworkisproperlycited. | 603 LUDENA URQUIZO Et AL. (average of 4.1%) compares favorably with those of wheat (2.7%) and corn (1.7%). Moreover, the amounts of calcium, magnesium, iron, and phosphorus (especially calcium and iron) are significantly higherthaninmostothercereals(Bhargava,Shukla,&Ohri,2006; Repo-Carrasco etal., 2003). Quinoa oil is rich in polyunsaturated fattyacidssuchaslinoleicandlinolenicacid,whichhavethepotentialtohelpindegenerativediseasessuchascardiovasculardiseases, cancer,inflammatoryandautoimmunediseases.Quinoagrainshave highconcentrationsofpolyphenolsandantioxidantssuchasα-and γ-tocopherol—compounds suggested to have anticarcinogenic and anti-inflammatoryactivities.Theyarealsoagoodsourceofvitamin C,E,andfolicacid(Bhargavaetal.,2006;Jancurova,Minarovicova,& Dandar,2009;Repo-Carrascoetal.,2003;Schoenlechner,Wendner, Siebenhandl-Ehn,&Berghofer,2010).Quinoapericarpscontainup to5%saponins,whichgiveabitterandastringenttaste(Vega-Galvez etal.,2010).Itisnecessarytowashouttheseundesiredcompounds beforequinoacanbeconsumed. Dueto the highnutritionalvalue,good agro-ecologicaladaptabilityandlowwaterrequirements,quinoahaslatelyreceivedalot of attention, and several projects on a sustainable production are ongoing, to improve nutrition and to increase food security and farmer income (Giuliani, Hintermann, Rojas, & Padulosi, 2012). Quinoacouldalsohavepotentialtodecreasetheriskoftype-2diabetesandcardiovasculardiseases,forexample,hypertension(Dixit, Azar,Gardner,&Palaniappan,2011;Ranilla,Apostolidis,Genovese, Lajolo,&Shetty,2009).Moreover,quinoaisasuitablesourceofproteinforvegetariansandvegansand,becauseitisgluten-free,itis alsoanalternativecerealforpeoplesufferingfromcoeliacdisease andgluten-allergyproblems.Thus,theuseofquinoaisnotonlyimportantindevelopingcountriesbutalsoinaffluentcountrieswhere thereisaneedtointroducenewandmorenutritiousfoodproducts thatcouldsubstituterefinedcarbohydrate-richgrainssuchaswhite riceandwheat. Quinoahasbeen traditionallyused as cooked forsalads, soups, porridgesandstews,asfriedpatties,anddrinks.Othermorerecent usesareasbreakfastcereals,granolabars,andbeer.Quinoacanalso bepoppedandextruded,andusedas“healthy”snacks.Grainscanbe milledintoflourandusedforbread-making,pasta,biscuits,andother processed foods (Ahamed, Singhal, Kulkarni, & Pal, 1996; Bhargava etal., 2006; Diaz etal., 2013; Giuliani etal., 2012). Although there aremanyquinoavarieties,thevarietymostlyexportedorcultivated inEuropenowadaysisthe“quinoaReal”.Itisacolorlessgrain(creamy), withthe largestgrainsize (2.2mm),andpreferredin the agro-food industryaroundtheworld.However,coloredquinoa(redorblack)varietiesareincreasinglyrequestedbecauseoftheirgoodorganoleptic potential. Inordertoexpandthetraditionalusesofquinoaandtoprovide new, healthierandmorenutritiousfoodproducts,this studyaimed atdevelopingaquinoa-basedfermentedbeverage.Thefermentation process during the preparation of the drink was monitored aswell asthemetabolicactivityduringthe fermentationand storagetime. Moreover,thepreliminaryacceptabilityofthedrink,byvoluntarypanelists,wasevaluated. 2 | MATERIALS AND METHODS 2.1 | Raw materials and nutritional composition Two different quinoa varieties, Rosada de Huancayo (RH) and Pasankalla(PK),bothcultivatedinJunín,Peru,werecomparedduring thestudy.PKwasoriginallyfromtheAndeanPlateauofPuno,Peru (4,000–5,000mofaltitude),buthasbeenadaptedtoloweraltitude inordertoimproveitsyield(Jancurovaetal.,2009).RHhasyieldsof 3–3.5t/haandaphysiologicalmaturityof170days.TheyieldforPK isalittlelower,3t/ha,buttheplantreachesitsphysiologicalmaturity inonly140days.RHisawhitevariety,whereasPKisaredvariety. Thenutritionalcomposition(moisture,protein,fat,crudefiber,and ash)ofthetwoquinoavarietieswasdeterminedbystandardmethods (AOAC,2005).Totalcarbohydratecontentwascalculatedbysubtractingthepercentagesumofmoisture,protein,fat,crudefiber,andash from100%.Saponincontentwasdeterminedbythestandardafrosimetricmethod(Koziol,1991).Briefly,quinoaseeds(0.5g)weremixed with5mlofdistilledwaterinacappedtubeandshakenvigorouslyfor 30s(fourshakes/s,upanddown).Aftera30minrest,thetubewas shakenvigorouslyagainfor30s.Thiswasdonetwiceandafter5min rest,foamheightwasread.Thepercentageofsaponinwasobtained fromthefollowingcalculation: 2.2 | Bacterial strains and culture conditions Threebacterialstrainswereusedasstartercultureforthefermentationofthequinoa-basedfermentedbeverage:Lactobacillus plantarum Q823, Lactobacillus casei Q11, and Lactococcus lactis ARH74. They wereselectedfortheirdiversetechnologicalandfunctionalproperties (RuizRodríguezetal.,2016),L. plantarumbeingapotentialprobiotic bacterium(Vera-Pingitoreetal.,2016).L. plantarum Q823 and L. casei Q11wereisolatedfromquinoagrains.L. lactisARH74isacommercial strain(ValioOy,Helsinki,Finland)wellcharacterizedasanexopolysaccharideproducer(Lehto&Salminen,1997).Overnightbrothculturesofthestrainswerepreparedthedaybeforethefermentation andincubatedat30°C.StrainsQ823andQ11weregrowninMRS broth(LabM,Bury,Lancashire,UK)andstrainARH74inM17broth (OxoidLtd.,Hampshire,UK). 2.3 | Samples preparation Quinoa seeds were separated from impurities (leaves, stones, etc.) and washed thoroughly to remove saponins (foamless). The seeds weresubsequentlydriedat60°Cforexactly8hr,reachingamoisture of2.26%forPKand3.72%forRH.Afterdrying,quinoaseedswere milled.Quinoaflourhadafinalparticlesizeofaround100μminboth varieties. For the processing of the fermented quinoa-based beverage, quinoa flours (each variety separately) were mixed with water at a concentration of 15% (w/v) (the experimentally selected minimum concentration required to prevent syneresis). The resulting quinoa %saponin=0.646[heightoffoam(cm)]−0.104∕sample�sweight(g) ∗10 604 | LUDENA URQUIZO Et AL. slurries(250ml)werethengelatinized/pasteurized(95°Cfor10min) andcooleddowntoambienttemperaturebeforethestartoffermentation.All three bacteriawere inoculated at a concentration of 1% andsamplesweresubsequentlyfermentedfor6hrat30°C(Figure1). Afterfermentation,100mlsampleswerestoredat5-7°Cfor28days. Threebiologicalreplicatesweredoneforeachsample. The pH, Total Titratable Acidity (TTA), and viscosity (Rotary ViscometerPCR-RVI3,Model20,UK)ofthesamplesweremonitored beforeandafterfermentation,andat1,12,and28daysofstorage. TTAwastitratedwith0.1mol/LNaOHtoafinalpHof8.5,detected byapHmeter.TTAwasexpressedasmlof0.1mol/LNaOHneeded toachievepH8.5. 2.4 | Culture viability determination Todeterminethefermentationcapacityofthebacterialstrainsand theirviabilityduringthestoragetimeoftheproduct,bacterialcounts weremeasured.Samplesweretakenbeforeandafterfermentation, andat1,12and28daysofstorage.Bacterialnumbersweredeterminedbyplating0.1mlofthreeappropriatedilutions,induplicate,on MRSagar(LabM)plates.MRSagarplateswereincubatedat30°Cfor 2daysandtotalbacterialcoloniescounted.Moreover,eachbacterial strainwasmonitoredseparatelybasedontheirmorphologicalcolony differentiation:ThecoloniesfromL. plantarumQ823arebigwhiteand shiny;thecoloniesfromL. caseiQ11haveirregularborders;andthe coloniesfromL. lactisARH74aresmall,opaque,flat,andtranslucent. 2.5 | Metabolic activity during fermentation Themetabolicactivityofthefermentedbeveragewasmonitoredby measuringtheamountsofglucose,maltose,andsucrose,aswellas thelactic,acetic,andmalicacid,beforeandafterthefermentation, andat1,12and28daysofstorage.Allanalyseswereperformedwith capillaryzoneelectrophoresis(CE). BeforeCEanalysisofsugars,2gofhomogenized,unfrozen(3hr in room temperature) sample was diluted with 6–7ml of ultrapure waterandmixedthoroughly.Aquantityof100μlofCarrezreagentI (potassiumhexacyanoferrate(II)trihydrate,MERCKKGaA,Darmstadt, Germany)andCarrezreagentII(zincsulfateheptahydrate,J.T.Baker, Deventer,Netherlands)wereaddedtothesuspensionandthepHwas adjustedto7–8with0.1mol/Lsodiumhydroxide(NaOH,J.T.Baker). Themixturewascentrifugedat3000rpm,for2minatroomtemperature. Subsequently, the supernatant was collected and adjusted to 10mlwithultrapurewater,followedbyfiltrationthrougha0.45μm syringefilter(VWRInternational,Darmstadt,Germany). Fortheanalysisoforganicacids,theextractswerepreparedina similar fashion except that the Carrez reagents and subsequent pH adjustmentswereomitted. TheCEinstrumentusedwasP/ACEMDQcapillaryelectrophoresis systemby Beckman CoulterInc. ( Fullerton,CA, USA)with a diode arraydetector.TheUVdetectionwasat270nmandindirectdetectionat232nmforsucroseandorganicacids,respectively.Sugarswere measuredwithmodifiedmethodofRovio,Yli-Kauhaluoma,andSiren (2007)applyingbuffersolutionof130mmol/LNaOHand36mmol/L disodium hydrogen phosphate (Na2HPO4, MERCK) at pH 12.6.The separationswere undertaken at 16°C in uncoated fused-silica capillarywith I.D.of25μmand total length of 40cm (effective length of30cm).Separationvoltagewas12kV.StandardsolutionsofD(+)- sucrose (VWR), D(+)-maltose (Sigma-Aldrich, Steinheim, Germany), D(+)-glucose (VWR), and samples were introduced to capillarywith pressureinjectionof0.5psifor10s. BuffersystemofBIS-Tris/Pyridinedicarboxylicacid(pH6.5)and Tris/Pyridine dicarboxylic acid (pH 8.1) by Analis along with their method (CEofix KIT, Anions 8) were used to analyze organic acids. Uncoated fused-silica capillary with I.D. of 75μm and total length of 60cm (effective length of 50cm) at a temperature of 20°Cwas employed. Before runs, capillary was conditioned with the buffers and flushed after with NaOH and ultrapure water. Separation was achievedwithvoltageof30kVusingreversedpolarity.Pressureinjectionof0.5psifor5swasappliedforsamples,andstandardsolutions offormicacid(ACROSOrganics,Geel,Belgium),L(-)-malicacid(Fluka, FIGURE1 Protocolfortheelaborationofthefermentedquinoa- basedbeverage | 605 LUDENA URQUIZO Et AL. Sigma-Aldrich, St. Louis, USA), and acetic acid (Supelco, Bellefonte, USA)wereusedforquantitativeanalysis. AllCEanalyseswereperformedastriplicatesusingL(+)-arabinose (Fluka)asaninternalstandardforsugarsandquinicacid(MERCK)as aninternalstandardfororganicacids. 2.6 | Preliminary organoleptic acceptability of the final product Thefinalproductswereevaluatedby20volunteers.Fourdifferent productswereevaluated:RH,PK,RHmixedwithbilberryjam(20% bilberryand3%ofsugar),andPKmixedwithdarkchocolate(12%, Fazer Oy, Helsinki, Finland). The additives were chosen according tothemostsuitablefinalcolor.Theacceptabilityoftheappearance, color,texture,andflavorwasexpressedbyahedonicscale(Nicolas, Marquilly,&O’Mahony,2010). 2.7 | Statistical analysis The experimental data were evaluated using analysis of variance (ANOvA)andTukeytest,bothconsideringasignificancelevelofp < .05. TheanalyseswereperformedwithstAtgRAphIcs cENtURIONXVsoftware (StatPointTechnologies,Inc.,Warrenton,VA).Alltheanalyseswere carriedoutintriplicatesexceptforthepHandTTA(duplicates). 3 | RESULTS AND DISCUSSION 3.1 | Nutritional and functional differences between quinoa varieties The superior nutritional value of quinoa compared to many other cereals or grains is well documented (Bhargava etal., 2006; Comai etal.,2007;Vega-Galvezetal.,2010).However,thenutritionalcompositionofquinoavarietiesmaydifferconsiderably(Repo-Carrasco- Valencia, Hellstrom, Pihlava, & Mattila, 2010). Although adequate comparativestudiesin this respect haveapparently not been done yet,itisknownthatthenutritionalcompositionofquinoavarieties isinfluencedbystronggeneticvariability,environmentalandclimatic factors(Gonzalez,Konishi,Bruno,Valoy,&Prado,2012).Inthisstudy, thenutritionalvalueoftwoquinoavarietieswasstudied.QuinoaPK showedsignificantly(p < .05)highercontentofproteinandfiber,and lowercontentoftotalcarbohydratesandsaponinthanRH,whereas thefatcontentwassimilarbetweenboththevarieties(Table1). The quinoa flourswere mixed withwater at a concentration of 15% (w/v). This concentration was assessed beforehand (data not shown)anditwastheminimumconcentrationrequiredtoobtaina drinkwithout syneresis(waterphaseseparation) duringthestorage time for both varieties. However, the viscosities of the flour-water slurriesweresubstantiallydifferentinthebeginningoftheprocess. Quinoa’sstarchgranuleshaveverygoodpastingpropertiesandcan beusedtoproducehigh-viscositydough.Quinoastarchhasalsoexcellentstabilityunderfreezingandretrogradationprocesses(Abugoch James, 2009; Ahamed etal., 1996). However, differences between quinoavarietiesarenotwellstudied.Inourstudy,beforefermentation,theRHdrinkwassignificantly(p < .05)moreviscousthanthePK one,whichwasnotexpectedduetothefactthathigherproteininPK isknowntoleadtoaharderandmorecohesivetexture(Wu,Morris, &Murphy,2014).Afterfermentation,althoughtheviscositywasreducedwithbothquinoavarieties,theeffectwasmoreprominentwith RH.Consequently,bytheendofthestoragetime,bothquinoavarietieshadthesameviscosity(Table2). The pH decreased andTTA increased significantly in the drinks asaresultofthefermentation.FermentedbeveragesrequireacidpH (4.0–4.5) in order to survive storage (Gupta, Cox, & Abu-Ghannam, 2010). No significant change was seen during the storage time althoughtherewasaslightdecreaseinpHandproportionalincreasein TTA(Table2).Similarbehaviorwasobservedinoat-basedfermented beverageswherethesefactordynamicswerelow(Angelov,Gotcheva, Kuncheva,&Hristozova,2006;Guptaetal.,2010).Betweenquinoa varieties,therewasnostatisticalsignificantdifferenceinpHandTTA. Theseresultsshowedthatnotonlyquinoavarietieshavesignificantlydifferentnutritionalcontentsbutthatthefloursbehavedifferentlywhenslurriedinwater.Thesedifferenceswillbestudiedinmore detailedinthefuture. 3.2 | Bacterial strains, growth, and viability Fermentationas a foodprocessing technique is not a novel procedure.However,fermentationwithproperstarterculturescanreduce theuseofartificialadditivessuchasstabilizers,thickeners,orflavors (Tiwari,Norton,&Holden,2013).Fermentationbyknownlacticacid bacteriawasusedfortheelaborationofthequinoa-basedfermented beveragereportedherein.Theinoculationof1%(v/v)ofeachbacterialstrainintothequinoabeverageresultedininitialbacterialcounts ofapproximatelylog8CFU/ml(Figure2).After6hrat30°C,these bacteriawereabletogrowtoaleveloflog9.5CFU/mlanddecrease thepHtoaround4.Duringthestorageperiod,thebacteria(withthe exceptionofARH74)provedtobequitestablewithevensomeincreaseinnumbersinthePKbeverage(Figure2).Aftera28-daystorageperiod,L. plantarum Q823 and L. casei Q11 were detected at levels TABLE1 Nutritionalcompositionoftwoquinoavarieties(asg 100gdrymatterand%) Quinoa varieties Rosada de Huancayo Pasankalla Moisture 10.52 ± 0.05a10.61 ± 0.00b Protein(N×6.25) 12.75 ± 0.01a14.08 ± 0.27b Fat 5.18 ± 0.12a5.07 ± 0.06a Crudefiber 2.70 ± 0.03a2.83 ± 0.07b Ash 2.51 ± 0.07a2.29±0.05b Totalcarbohydrates 66.35 ± 0.18a65.12 ± 0.33b Saponincontent 0.66%a0.00%b Theresultssharingthesuperscriptletter(withintherows)arenotsignificantlydifferent(p < .05). 606 | LUDENA URQUIZO Et AL. morethanlog9CFU/ml.PreviousstudieshavealsoreportedhighstabilitywithL. plantarumduringstorageat4°Cinoat-fermentedbeveragesfor21days(Angelovetal.,2006;Guptaetal.,2010).However, L. lactis ARH74 was lost during the storage time (data not shown). TheuseofPKorRHvarietydidnotsignificantlyaffectthegrowthor viabilityofthestrains. Thenumberofbacterialcellsin foodproductsthatclaim probioticpropertiesandthenumberassociatedwithsignificantoutcomes inclinicaltrialsareintherangeof1–10billionCFUperdose(Naidu, Adam,&Govender,2002;Reid,2005;Guarneretal.2012).Wehave previouslyreportedthatL. plantarumQ823cansurvivethepassage throughthehumanintestinaltractandthus,beapotentialprobiotic bacterium(Vera-Pingitoreetal., 2016).Moreover,the quinoa-based fermentedbeveragedevelopedandreportedhereinisabletoreacha L. plantarumQ823populationhigherthanlog9CFU/mlandthusbe ontherangeforhavingprobioticactivities.Consequently,thequinoa- basedfermentedbeveragehasthepotentialtobeusedasafunctional food even though the actual health benefits should be proven in a long-termhumanclinicaltrialinordertoclaimthatanyfoodproduct hasprobioticproperties. 3.3 | Metabolic activity Fermentationisafoodprocessingtechniquethatcanhelptoimprovetexture, structure, nutritionalvalue, staling rate, andshelf life of food products. These qualities are associated with the TABLE2 ChangesinthepH,totaltitratableacidity(TTA),andviscosityduringthefermentationandstorageofthequinoa-basedbeverage Days pH TTA* Viscosity (Pas) RH PK RH PK RH PK Mean SD Mean SD Mean SD Mean SD Mean SD Mean SD 0†6.47a0.07 6.47a0.07 2.20a0.10 2.25a0.15 53.79a11.47 28.48a1.89 0.25‡4.20b0.20 4.39b0.01 7.70b0.20 7.30b0.10 36.76ab 16.27 21.76ab 2.74 14.09b0.11 4.28bc 0.08 8.25bc 0.25 7.90b0.10 25.90ab 17.71 17.49bc 3.93 12 3.84b0.16 4.14bc 0.04 9.25bc 0.25 8.35b0.45 14.97b10.31 12.73c4.26 28 3.86b0.15 3.97c0.07 9.50c0.50 8.60b0.60 10.62b5.20 10.59c2.05 *TTA,mlof0.1mol/LNaOHper10g. †Beforefermentation(0hr). ‡Afterfermentation(6hr). Theresultssharingthesuperscriptletter(withinthecolumns,inlowercase)arenotsignificantlydifferent(p < .05). FIGURE2 Stabilityandviabilityofthe bacteriainthequinoa-basedfermented beverage FIGURE3 Metabolicactivityinthe quinoa-basedfermentedbeverage | 607 LUDENA URQUIZO Et AL. production of organic acids, exopolysaccharides, aroma compounds, and antifungal compounds by lactic acid bacteria (LAB) (Wolteretal.,2014).Inthisstudy,sugarsandorganicacidswere monitoredduringfermentationandstoragetimeofthedeveloped foodproduct. The concentration of glucose, sucrose, and maltose before fermentationshowsthattherearesignificantdifferencesbetweenthe quinoavarieties(Figure3).GlucoseconcentrationinRHwas7.4mg/g, whereasinPK,itwas9.0mg/g.Glucoseconcentrationsdecreasedin bothvarietiesduringthestoragetimetolevelsof5.2and8.0mg/gin RHandPK,respectively.Sucroseandmaltosewerenotsignificantly differentbetweenquinoavarieties(Figure3). There was a rapid increase in lactic acid concentration but not inaceticormalicacidduringthefermentation(Figure3).Duringthe storageperiod,lacticacidcontinuedtoincreasereachingconcentrationsof7.3and7.5mg/ginRHandPK,respectively. Althoughendogenousenzymesfromtheflourscaninfluencethe metabolicactivityofthedrinks,thedecreaseinglucoseandincrease inlacticacidwasmostlikelyduetotheactivityofthelacticacidbacteria.Surprisingly,aceticacidwaspracticallyabsentindicatingthatthe fermentationoftheproductwasmostlyhomolactic.Itis,however,unknownthewidedifferenceinglucoseconcentrationbytheendofthe storagetimeinbothvarieties,especiallybecausetheendlacticacid concentrationwasalmostthesame.Thisinformationwillbestudied moredeeplyinthefuture. 3.4 | Preliminary organoleptic acceptability of the final products Theorganoleptic acceptability offour final products was evaluated (Table3).RHandPKwerenotwellreceivedassuchduetothecharacteristicsourtaste.However,theacceptabilityofthesebeverages was very good when bilberries (Vaccinium myrtillus) and chocolate wereusedasflavorings.Therewerenotsignificantdifferencesinthe acceptabilityoftheproductmadefromPKflourcomparedtoRHalthoughthecolorofthefinalproductwasdark-brownandthusnotso appealing. Althoughthisorganolepticacceptabilitytrialwasverypreliminary anditisclearthatextensiveresearchshouldstillbedoneonsensory attributes,itshowsthatquinoa-baseddrinkshavethepotentialtobe wellreceivedbyconsumers. 4 | CONCLUSIONS A fermented quinoa-based beverage was successfully developed. RosadadeHuancayo(RH)andPasankalla(PK)canbothbeconsidered goodvarietiestobeusedinfoodprocessingwith special attention toPKduetoitshigherproteinandlowersaponincontent,itslower lossofviscosity,anditshighersugarcontent.Thedevelopmentofa foodproductbasedonfermentationprovideda“spoonable”beverage,withoutphaseseparationandsafelowpH,andtheseproperties werestableduringthe28-daystorageperiod.Theseproductscould beagoodsourceofprotein,fiber,vitaminsandminerals,makingthem notonlyagoodsnackforthecoeliacandlactose-intolerantpopulationbutalsoanewandexoticalternativetoconsumersingeneral. Moreover,they mightsupportthegrowth andviabilityofprobiotic bacteria,suchasL. plantarumQ823providedthatthestrainhasactual long-termhealthbenefits. ACKNOWLEDGEMENTS This work was part of the project “Improving Nutrition of Andean andAmazonianPopulation:Health-promotingBioactiveCompounds inAndeanandAmazonianFoodMaterialsandDiets”fundedbythe MinistryforForeignAffairsofFinland. CONFLICT OF INTEREST Theauthorsreportnoconflictofinterest. REFERENCES Abugoch James, L. E. (2009). Quinoa (chenopodium quinoa willd.): Composition, chemistry, nutritional, and functional properties. Advances in Food and Nutrition Research,58,1–31. Ahamed, N., Singhal, R., Kulkarni, P., & Pal, R. (1996). Physicochemical andfunctionalpropertiesofChenopodium quinoastarch.Carbohydrate Polymers,31,99–103. Angelov, A., Gotcheva, V., Kuncheva, R., & Hristozova, T. (2006). Developmentofanewoat-basedprobioticdrink.International Journal of Food Microbiology,112,75–80. AOAC. (2005). Official methods of analysis (18 edn). Gaithersburg, MD: AOACIntl. 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