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Factors Affecting Color Formation During Storage of White Crystal Sugar

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There are many reports of problems caused by incorrect storage of sugar production ‐ hardening, caking, dimming, losses and fires. Given the importance of crystal sugar, one of the main sugarcane‐based products in Latin America or around the World, there is a growing realization of work to increase efficiency and avoid losses in the manufacturing. However, during sugar storage, it is known that environmental factors (temperature, humidity, light and weather) influence the quality of the final product – several types of crystal sugars, such as: very high polarization (VHP); very‐very high polarization (V‐ VHP), and whites – Types 1, 2, 3 or 4. During storage, the temperature must be not exceed and/or sensitive to variations. The optimum relative humidity is 55‐65% with the maximum equilibrium moisture at 65%. This work aimed to assess which factors are important to accentuate the color during the storage process (specially, ICUMSA color and sucrose hydrolysis) of white sugar (Type 1). The results of the amounts of reducing sugars and ICUMSA color were evaluated by the response surface. It was concluded that there were changes in the amounts of reducing sugars, sucrose and ICUMSA color in the analyzed samples, that moisture is the most decisive factor. Therefore, it became clear that you should avoid such conditions of high humidity and temperature in storage sugar, in order to preserve the quality, which is highly perishable.
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doi:10.14355/fmfi.2015.04.001
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FactorsAffectingColorFormationDuring
StorageofWhiteCrystalSugar
ClaudioLimadeAguiar,AnaLauraBodoniRocha,JéssicaRodriguesJambassi,AntonioSampaio
BaptistaandRobertaBergaminLima
UniversidadedeSãoPaulo,ESALQUSP,DepartamentodeAgroindústria,AlimentoseNutrição,SetordeAçúcar
eÁlcool,LaboratórioHugotdeTecnologiaemSucroderivados,Piracicaba,SP.,Brasil.
Abstract
Therearemanyreportsofproblemscausedbyincorrectstorageofsugarproduction‐hardening,caking,dimming,lossesand
fires.Giventheimportanceofcrystalsugar,oneofthemainsugarcanebasedproductsinLatinAmericaoraroundtheWorld,
thereisagrowingrealizationofworktoincreaseefficiencyandavoidlossesinthemanufacturing.However,during
sugarstorage,itisknownthatenvironmentalfactors(temperature,humidity,lightandweather)influencethequalityofthe
finalproductseveraltypesofcrystalsugars,suchas:veryhighpolarization(VHP);veryveryhighpolarization(V
VHP),andwhitesTypes1,2,3or4.Duringstorage,thetemperaturemustbenotexceedand/orsensitivetovariations.The
optimumrelativehumidityis5565%withthemaximumequilibriummoistureat65%.Thisworkaimedtoassesswhichfactors
areimportanttoaccentuatethecolorduringthestorageprocess(specially,ICUMSAcolorandsucrosehydrolysis)ofwhite
sugar(Type1).TheresultsoftheamountsofreducingsugarsandICUMSAcolorwereevaluatedbytheresponse
surface.Itwasconcludedthattherewerechangesintheamountsofreducingsugars,sucroseandICUMSAcolorin
theanalyzedsamples,thatmoistureisthemostdecisivefactor.Therefore,itbecameclearthatyoushouldavoidsuch
conditionsofhighhumidityandtemperatureinstoragesugar,inordertopreservethequality,whichishighlyperishable.
Keywords
Sugar,Sucrose,Storage,Humidity,Temperature
Introduction
Theimportanceofindustryandtradebalanceofsugarcaneanditsderivativesintropicalcountries,especially
sugar,encouragepapersaddressingtheexportperformance.Thissectorhasdirectinvolvementinbothdomestic
andinternationalmarkets,sincesugarcaneisamajorcropintermsofarea,productionvolumeandcost(Alvesand
Bacchi,2004).Sucroseisthecarbohydratemoreinterestedintheprocessingofsugarcane,whichisdesiredin
crystallizedform,anditislikelythattheeffectoftemperature,enzymesandmicroorganismsismoreimportant
(Mantelatto,2005).
However,ABIA(2010)reportedthatduringstorageofcrystalsugar,itisknownthatdifferentenvironmental
factorsaffectthequalityofthefinishedproduct.Thesefactorescanbe:temperature,humidity,presenceoflight,
andthetime,i.e.,theperiodwhentheproductisstored.AccordingtoLegendreandClarke(LegendreandClarke),
thesugarcanejuicecolorandthereforesugaroriginatefromvariouscompounds,suchasflavonoids,phenolic
compounds,andthesepigmentsthatreactwithreducingsugars,whichaffectdirectlythejuicecolorandsugar
quality.Theformationofcoloredcompoundsintheprocessiscarriedoutmainlybydegradationofsugar(sucrose)
andtheformationofthemonosaccharides,glucoseandfructose(Mónicaetal.,2004).
Ingeneral,browningreactionsaredetrimentaltothenutritionalvalueofthefoodinquestion,andmayoccur
duringprocessingandstorageoffoodstuffs.Itisthereforenecessarytofindtheconditionstopreventthese
reactions,therefore,notonlytopreventanychangeindiet,butotherchangesthatmaycausethefood
unacceptablefortheconsumer(Eskinetal.,1971).
Whenrelatedtoinadequatestorage,formingadarkerisstillavisualthemethatcaninfluencethebuyingdecision
oftheconsumertooptforasugarmorewhite.Inaddition,changesintheenvironmentcausechangesintosugar,
becauseitabsorbsmoistureforbalance,andthismoisturedissolvesasmallamountofcrystalsugar.However,
whenthereisadecreaseinhumidity,sugarbecomesdifficultduetowaterevaporationandrecrystallizationofthe
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molecularsucrose.Thecontactpointoftwocrystalsexpandssothattheyholdtogether.Eachtimethisprocess
occurs,sugarturnsintoahardmassthatsignificantlyaffectsthequalityoftheproduct(Howes,1966).According
toSRI(2007),itisgenerallyacceptedthatforevery10ºCriseintemperaturetherateofcolourformationincreases
byafactorofapproximatelythree.AccordingtoTIS(2015),duringstorage,thetemperatureofsugarsmustnot
exceedamaximumand/ortheyaresensitivetotemperaturevariations,andfortheoptimumrelativehumidityis
5565%withthemaximumequilibriummoistureat65%.
Inthiscontext,thisstudywasaimedatassessingthedeterminantsthatemphasizetheprocessofcolorformation
duringstorageofwhitecrystalsugarandhowtheinteractionbetweenthemare,allowingknowledgetoadaptthe
warehousescurrentlyusedtomaintainproductquality.
Material and Methods
SamplePreparationandExperimentDesign
AssayswereperformedatHugotSugarTechnologyLaboratoryfromtheDepartmentofAgriFoodandNutrition
(LAN/ESALQ)UniversityofSanPauloandsamplesofhighpuritysucrosewereused(99%;SynthCo.,SanPaulo,
Brasil).
Toevaluatecolorchangesinthecrystalsugarsamples,thesampleswereplacedinsealeddesiccators,wherethe
relativehumiditywascontrolledwithinthevesselviasaturatedsolutionsof:magnesiumchloride(MgCl2.6H2O)to
obtainequalto30%moisture;magnesiumnitrate(MgN2O6.6H2O)forhumidityequalto50%;andsodiumchloride
(NaCl)forhumidityequalto70%(Rockland,1960).Theassembledgroupwastakentoafurnacewithcontrolled
temperatureat30,40and50°Cfor6,12and24hoursforaccompanyingfactorspotentiallyaffectingtheformation
ofcolorduringstorage,beingthefactorsstudied:temperatureandhumidity,bothfactorsassociatedwith
increasingintervalsoftime.
Forafullfactorialexperimentaldesignused23experimentswith3atthecenterpointreplicates,withatotalof11
trials(Table1).
TheresultsevaluatedthefactorsthatinfluenceICUMSAcolorformationincrystalsbytheresponsesurface
methodologywithallanalyticaldataevaluatedinstatisticalpackageStatSoft(2001),andthedataweresubjectedto
analysisofvariance(ANOVA)usingtheFtestandtheaveragescomparedbyTukeytestat5%probability(p
<0.05).
TABLE1.FACTORS(INPARENTHESES)ANDFULLFACTORIALEXPERIMENTALDESIGNDOMAIN23FOREVALUATINGFACTORSTHATAFFECTTHESUGAR
STORAGE.
Runsfactors
x1:humidity(%)x2:temperature(°C)x3:time(hours)
1()30()30()6
2(+)70()30()6
3()30(+)50()6
4(+)70(+)50()6
5()30()30(+)24
6(+)70()30(+)24
7()30(+)50(+)24
8(+)70(+)50(+)24
9(0)50(0)40(0)12
10(0)50(0)40(0)12
11(0)50(0)40(0)12
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EvaluationofReducingSugarsandICUMSAColorintoCrystalSugarSamples
Theparametersexaminedwere:
(1)Reducingsugars(RS)bytheSomogyiNelson(Nelson,1960)colorimetricmethod,inwhichthereadingwas
takentransmittance(T%)at520nminspectrophotometerUVMini1240(ShimadzuCo.,Kyoto,Japón).Reducing
sugars(mgg1)werecalculatedbythefollowingequationofthestandardcurveofglucose:
  %.
.
0.00157
(2)ICUMSAcolor:themethoddescribedbyLopes(1985),inwhichthesampleswerereadinspectrophotometer
UVMini1240(ShimadzuCo.,Kyoto,Japón)at420nm.Distilledwaterwasusedascontrolsolutiontoevaluatethe
device.Thecolorindexwascalculatedaccordingtothefollowingequation:
  1000   log
bc
T=solutiontransmittance(%);b=measureofthecell(cm);c=solutionconcentration(gmL1);
AnalysisofSugarsUsingUltraFastLiquidChromatograph(UFLC)
ThemethodemployinganUFLCchromatographicsystem(ShimadzuCo.;Kyoto,Japan)equippedwithELSLT
(evaporativelightscatteringatlowtemperature)detectorwascarriedoutat35ºCusingisocraticelutionof
acetonitrile(HPLCgrade;TediaCo.,Farfield,USA)andwater(deionized;Millipore,France)ataflowrateof1.0
mLmin1.Isocraticelutionwasemployedfor12minwithamixtureof70:30(v/v)acetonitrilewater.Nitrogen
(99.0%;AirLiquide,SãoPaulo,Brazil)at350kPawasusedtonebulizetheeffluentcomingfromthecolumn
NH2P504E(250mmx4.6mm)ShodexPacked(ShodexGroup,Kawasaki,Japan)at30ºC,andtheevaporation
temperatureofthechromatographiceluentwas30°C.Beforetheinjection(samplevolume=10μL),thesamples
wereclarifiedwithleadsubacetate,dilutedto1/10(v/v),andfilteredthroughDuraporefilters0.45μmand13mm
(MilliporeMerck,SãoPaulo,Brazil).Allsugars(sucroseGainat3;andglucoseandfructoseGainat7)were
analyzedagainstknownstandardspurchasedbySigmaAldrich(99.0%,MSgrade).
UVvisibleSpectrophotometricAnalysisoftheCrystalSugarSamples
Sampleswereconditionedindesiccatorswithrelativehumiditycontrolled,suchas30,50,and70%,suchas
temperaturesof30,40,and50ºC.Thestoragetimewas6,12,and24h,accordingtoTable1.Toevaluatingthe
spectraofmaximumabsorption(scanning),thesampleswereadded12.5gofsugarandthendilutedindistilled
water(25mL).ThespectrawereobtainedbyUVMini1240spectrophotometer(ShimadzuCo.,Kyoto,Japan)
between250and470nmatdifferenttimeintervals,being:0,24,and96hafterstorage.Thesampleswereexamined
underUVvisiblelightforproximateanalysis.ForUVVISspectrophotometeranalysis,crystalsugarswere
conditionedindesiccatorswithrelativehumiditycontrolled,suchas30,50,and70%,suchastemperaturesof30,
40,and50ºC.Thestoragetimewas6,12,and24h,accordingtoTable1.Toevaluatingthespectraofmaximum
absorption(scanning),thesampleswereadded12.5gofsugarandthendilutedindistilledwater(25mL).The
solutionswerescannedinthewavelengthrangingfrom2001100nmusingUVMini1240spectrophotometer
(ShimadzuCo.,Kyoto,Japan)andthecharacteristicpeaksweredetected.ThepeakvaluesoftheUVVISwere
recordedintriplicate.
Results and Discussion
ByFig.1a,itcanbeinferredthatthereisaprogressiveincreaseinthelevelsofreducingsugarswithtemperature
change.Thismayberelatedtotemperatureincrease,contributingtotheprocessofinversionofsucroseinto
glucoseandfructose(evaluatedasreducingsugarsbySomogyiNelsonmethod).Fig.1banalysisallowedverifying
that,sincethereisnoincreaseinrelativehumidity,obtainedhigherconcentrationsofthereducingsugars,thusthe
reactioniscatalyzedasahigherwatercontentavailableintheenvironment(desiccators).Inthisexperiment,the
levelsofreducingsugarsincreasedastemperaturealsoincreased.
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Rodriguesetal.(2000)instudiesofcatalytichydrolysisofsucrosefoundthatthemostinfluentialfactoristhe
temperature,andthatthisincreasewasdirectlyproportionaltotheinversion(orhydrolysis)ofsucrosetoglucose
andfructose.
FIG.1.RESPONSESURFACEGENERATEDBYTHEINTERACTIONFROMTHEVARIABLES(A)TEMPERATURE(°C)ANDTIME(H)AND
FORINTERACTIONFROMTHEVARIABLES(B)MOISTURE(%)ANDTEMPERATURE(°C)TOOBTAINVALUESREDUCINGSUGARS
(RS)(mgg
1
).
Afterthemelting,sucroseloseswaterandglucoseandfructoseanhydridesorglucosansandlevulosansbecome.
Thereactionisselfcatalyzedwaterformedasafunctionofacceleratingthereaction.Theanhydridesformed
combinewithwatertoproduceacidderivativesandhydrolyzetheremainingsucrose,fructoseandglucose.The
glucosansandlevulosansformedalsocanbecombinedwithwaterandreappearfructoseandglucose(Oettereret
al.,2006).
AccordingtoAraujo(1995),sugarsattemperaturesabovedescribed120°Carepyrolizedatvariousdegradation
productsandhighmolecularweightcalleddarkcaramel.Thisreactioninvolvesthesugardegradationinthe
absenceofaminoacidsorproteins.
Duringthewholetimeofdehydrationandhydrolysisreactionsoccurring,withpredominanceofacidssuchas
aceticandformic,oraldehydessuchasformaldehydeand5hydroxymethylfurfural,diacetyl,carbonylandenol
groups.Thesecompoundsareresponsibleforthearomaandcolor,whichrecombineandformpolymerscalled
melanoidins(Oettereretal.,2006).
EvaluatingthetrendofincreasingICUMSAcolor(Fig.2),itappearsthatunderhighrelativehumidity,regardless
ofthetemperatureinquestion,therewasanincreaseincolor,withthefactthatstorageprocessofsugarwas
unwanted.
Fig.2cshowstheresponsesurfaceforICUMSAcolorfrominteractionbetweenhumidityandtime.Itwasnoted
that,regardlessofthetimeinexperimentaldesign,ICUMSAcolorwasmorepronouncedinthemaximumrelative
humidityused.ICUMSAcolorresultspresentedaninversebehavior,andseveralauthors(Arenaetal.,2001;Ibarz
etal.,2000;SapersandHicks,1989;Namiki,1988)haveshownthatthetimetemperaturerelationshipismarkedly
importantforthehydrolysisofsucrose.However,theformationofpigmentedcompoundspromotessugar’scolor.
Byanalyzingresponsesurfaces,onecanobserveageneraltrendofincreasingICUMSAcolor,andreducingsugars
accordingtoprogressiveincreasesinrelativehumidityoftheenvironment,regardlessofthetemperaturesand
timesused.
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FIG.2.RESPONSESURFACEGENERATEDBYTHEINTERACTIONFROMTHEVARIABLES(A)TEMPERATURE(°C)ANDHUMIDITY(%);
FORINTERACTIONFROMTHEVARIABLES(B)MOISTURE(%)ANDTIME(H);ANDFORINTERACTIONFROMTHEVARIABLES(C)
TEMPERATURE(ºC)ANDTIME(H)TOOBTAINVALUESICUMSACOLOR.
Belowrelatedtablesofanalysisofvariance(ANOVA)fortreatmentsperformedinthesamples.InTable2,wecan
observethedataforanalysisofICUMSAcolorandTable3observesthedataforanalysisofreducingsugars.
TABLE
2.
RESULTSOFANALYSISOFVARIANCEFORICUMSACOLOR
.
 SSdfMSFp
Regression16579,126862763,1886,347490,047578
Residual1741,27894435,3197
Total18320,4057101832,041
 0,904954
TABLE
3.
RESULTSOFANALYSISOFVARIANCEFORREDUCINGSUGARS
.
 SSdfMSFp
Regression573980,976,0095663,4914,90870,010375
Residual25666,494,006416,621
Total599647,4510,0059964,75
0,957197
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AsshowninTables2and3,valuesweresignificantatthe5%levelofconfidenceforallanalyses.Therefore,the
responsesurfaceresultsarepresentedbelow.Throughdataanalysis,itcanbeinferredthatthereisaprogressive
increaseinthereducingsugarcontentswithincreasingoftemperature.Thisfacthasbeenrelatedtothetrendof
increasingtemperature,whichcontributedtotheprocessofinversionofsucrosetoglucoseandfructose.
Echavarriaetal.(2013),instudiesofnonenzymaticbrowninginpotatos,reportedatrendinreductionofsucrose
levelsforincreasingtemperature.
AccordingtoTomasiketal.(1989)andSuarezPereiraetal.(2010),intheoligomerizationreaction,thereisa
formationofbrownandthechangeinmaterialtextureismoreviscous.First,theindividualsugarsreacttoforma
moleculecontaininganewformoftworingsconnectedbyathirdcentralring.Thiscompoundcanalsoreactby
threeways.Atfirst,waterlosingmoleculesformacompositestructurecalledcaramelans(C
12
H
12
O
9
),addingto
formsmalldarkparticlesofthesize0.46μm.Thesecondtypeofmoleculecanform,iscaramelens(C
36
H
18
O
24
),
whichaggregatetoformmoleculeswith0.95μm.Finally,itispossibletoformthecaramelins(C
24
H
26
O
13
),the
combinationoftwofructosedianhydridesandtheeliminationof27watermolecules.Thesecaramelinsare
generallyaggregateswithasizeof4.33μm.
Fig.3(adaptedfromTomasiketal.,1989)showsthedecompositionreactionofsucrose(seeFig.4)andthe
oligomerizationofglucoseandfructoseformedafterinductionbytemperatureandhumidity.
Theanalysisofthechromatograms(Fig.4)showsthattheyareconsistentwiththecharacteristicsucroseprofiles,
andthevariationintheglucoseandfructosecontentsresultsfromthehydrolysisofsucrose.Thedegradationof
sucrosewasrepresentedinFig.4.8.Sampleswerepreparedatconcentrationsof1gmL
1
andsubjectedto
chromatographicanalysisshowingreducedlevelsofsucrose.
FIG.3.CARAMELIZATIONREACTIONCAUSEDBYTHESUCROSEDECOMPOSITIONANDOLIGOMERIZATIONOFTHEGLUCOSE
ANDFRUCTOSE(ADAPTEDFROM20).
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FIG.4.CHROMATOGRAMSPROFILESOFSUCROSESUBMMITEDTODIFFERENTTREATMENTS(TABLE1)OFCRYSTALSUGAR
SAMPLES.
AccordingtoPalashudinSketal.(2012),instudieswithcaramels,itwasobservedthatthesucroseprofilewhen
subjectedtothemaximumabsorptionspectrumwasfoundtobeconsistentwithresultsobtainedinthis
experiment,whichdemonstratesthatchangesinthecurvesarerelatedwithformationofothercompoundsformed
duringthetreatmentandthegreaterthetimewas,thelargerthesechangesalsoare.Thatis,samplesforthechange
intheprofileofthemaximumabsorptionspectrumbetween250and470nmfordifferenttemperaturesandtime
intervalsareshowninFig.5(to30%,50%,and70%humidity).
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FIG.5.SPECTRASCANNINGBETWEEN250TO470nmOFTHESUGARSAMPLESUNDERDIFFERENTSTORAGECONDITIONS.
AnalyzingFig.5,itispossibletoobservechangesinabsorptionprofilesofthescanspectrumsamples,andithas
beenassociatedbyothercompoundssuchasquinones,caramelans,caramelensandcaramelins(PalashudinSket
al.,2012;Zhangetal.,2015;GolonandKuhnert,2013;GolonandKuhnert,2012),whichalsochangethecolorofthe
samplesandareresponsibleforthecharacteristiccaramel.Thereisaclearobservationthatthehigherthereaction
timeinthiscasewasanalyzedintime0,24and96h,thegreaterthechangeintospectraprofiles;andthehigherthe
temperaturewas,whichwastheformationofpigmentedcompounds,themoresamplesweremarked,.
Caramelization,accordingtoAraujo(1995),requiresneitheroxygennornitrogencompounds,occurringatthe
optimumpHof3.0and9.0,theproductionofcaramels.Nonenzymaticbrowningmayalsobetheoxidation
reactionofascorbicacidwhichrequiresoxygen,butdoesnotrequirenitrogencompoundsproducedbetweenpH
3.0and5.0toproducemelanoidins.Themechanismofthisbrowningreaction,accordingtoSeravalliandRibeiro
FocusingonModernFoodIndustryVolume4,2015www.fmfijournal.org
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(2007),isstillunknown.Accordingtotheauthors,itisknownthatheatingcausesbreakageofglycosidicbonds,
andformationofnewglycosidicbonds,andtheformationofunsaturatedpolymers‐caramels.
Morespecifically,Evangelista(2005)reportsthatthebrowningistheresultofthereactionbetweensugars
containinghydroxylandcarbonylgroups.Thisreactiontakesplaceathightemperatureswithsugardehydration
andformationofveryaldehydessuchas5hydroxymethylfurfural(5HMF)responsibleforthecharacteristicodor
ofcaramelizedsugar.
Conclusion
Theanalysisoftheeffectsofdifferentsugarstorageconditionsshowsthatthereareconsistentalterationsinthe
profileofsucrose,glucoseandfructosecontents,andthereiscolorformationduringthestorage.Bythemaximum
absorptionspectraitwaspossibletoobservechangesinabsorptionprofilesofthesamplessubjectedtohigh
temperaturesandhumidity,thusbeingthebinomialtemperaturetimeassociatedwithmoisture,importantfactors
inmaintainingthequalityofsugarwhitestored.
Acknowledgements
TheauthorsacknowledgethesupportoftheFoundationfortheSupportofResearchintheStateofSãoPaulo
FAPESP2009/546351)andNationalCouncilofScientificandTechnologicalDevelopment(CNPq506328/20104)
forfinancialsupportofthisresearchproject.
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