939KKU Res. J. 2012; 17(6)

Study of Volatile Compounds in Thai Rice Wine (Sato) Produced from Wheat

Thanut Amatayakul1*, Numchok Somsap2, and Premsiri Rotsatchakul2

1Faculty of Agricultural Product Innovation and Technology, Srinakharinwirot University, 10110, Thailand2Food Technology Program, Mahidol University, Kanchanaburi Campus, Kanchanaburi, 71150, Thailand*Correspondent author: tanut@swu.ac.th

Abstract

Metabolismofnitrogensourcesuchasaminoacidsisanimportantfactorcontributingtotheproduction

ofvolatilecompoundsinalcoholicbeverages.Inricewineproduction,aminoacidscomefromtheproteolysis

ofriceduringkojifermentation.ThisresearchstudiedvolatilecompoundsinSatoasaffectedbyrawmaterial

containinghighnitrogencontentsuchaswheatrice(WR)comparingwithglutinousrice(GR,controlsample).

Totalsolublesolid(TSS)andtotalnitrogen(TN)weredeterminedinkojiattheendoffermentation(7days).

Freeaminoacid,ethanolandvolatilecompoundsweremeasuredevery2days.TheTSSvaluesofkojimade

usingWRandGRwere17.13and21.53oBrix,respectively.TheTNvaluesofthosesampleswere0.240%and

0.085%,respectively.Duringalcoholicfermentation,higherreductioninTNwasobservedintheWRsample

(0.09%reduction)comparedwiththecontrolsample(0.04%reduction)duringthefirst2days,anditsTNcontent

continuouslydecreaseduntiltheendoffermentation.Thisinverselycorrelatedwithanincreaseinalcoholpattern

of the twosamples.At theendoffermentation, therewere12.7and10.8%(v/v)alcohol in theWRandGR

samples,respectively.Higherconcentrationof3-methyl-1-butanol,phenylethanol,phenetylacetateanddiethyl

succinatewasobservedintheWRsamplecomparedwiththeGRsample.However,someofthesevolatileswere

observedatthefirstdayofalcoholicfermentationsuggestingthatthesecompoundsarealreadyproducedduring

kojifermentation.

Keywords: Sato;Aminoacids;volatilecompounds

KKU Res. J. 2012; 17(6):939-949http : //resjournal.kku.ac.th

940 KKU Res. J. 2012; 17(6)

1. Introduction

Flavor is one ofmany important quality

characteristics of alcoholic beverages.Both volatile

and non-volatile compounds contributed to flavor

characteristics.Manyfactorshavebeenshowntoaffect

the productionof volatile compounds.Garde-Cerdan

andAncín-Azpilicueta(1)observedthatwinefermented

from nitrogen deficient must supplemented with

aminoacidsresultedinwinewithincreasedtotalesters,

isoamylacetateandphenylethylacetate.Thestudyby

Hernández-Orte,Ibarz,CachoandFerreira(2)showed

thatwineproducedfrommustwhichtheconcentration

of amino acids was doubled had fruity and fusel

characteristics.Theoretically,aminoacidsaremetabolized

to higher alcohols byyeast throughEhrlich pathway

(3,4).Hernández-Orte, Ibarz,Cacho andFerreira (5)

found that isoamyl alcohol, isobutanol and b-phenyl

ethanolwereproducedalongalcoholfermentation.In

addition, they reported that therewas lowereffectof

aminoacidonproductionofisoamylalcoholcompared

withb-phenylethanol.Thelaterwasespeciallyobvious

whentheconcentrationofphenylalaninewashigh.Esters

canbeproducedfromeitherthroughenzymaticreaction

(alcoholacetyltransferases)duringprimaryfermentation

aswellaschemicalesterificationbetweenfattyacidsand

alcoholsduringwineaging(6).

Due to the fact that higher alcohols are a

precursorinvoledinthesynthesisofester,metabolism

ofaminoacidsbyyeastwouldaffectconcentrationof

esterinalcoholicbeveragesanditsflavor.Inricewine

production, amino acids come from the proteolysis

of rice by acid proteases.These acid proteaseswere

producedduringkoji fermentation (7).However, the

aminoacidsdigestionoccurredmainlyduringalcohol

fermentation(8).Therewereseveralstudiesabouteffect

ofammoniumandaminoacidadditiononfermentation

kinetic andproductionof volatile compounds during

wine fermentation (2,5,6,9).However, only limited

informationhasbeenfoundfortheeffectofaminoacid

onproductionofvolatilecompoundsinricewine.As

inthecaseofwine,metabolismofaminoacidsyields

positiveresultsofhigheralcoholaswellasestersformation.

Therefore,theuseofrawmaterialthatishighinprotein

contentotherthanriceorglutinousrice,incaseofThai

ricewine,shouldyieldproductwithbettervolatileflavor.

Thisstudyinvestigatedvolatilecompounds,particularly

higher alcohols and acetate esters, inSato (Thai rice

wine) as affected by rawmaterial containing high

proteincontentsuchaswheatrice(WR)comparingwith

glutinousrice(GR)whichisusedasacontrolsample.

2. Materials and Methods

2.1 Experimental plan

Thairicewine(Sato)wasmadeusingeither

glutinous rice orwheat as rawmaterial.Aspergillus

nigerTISTR3257(MicrobiologicalResourceCentre,

Thailand Instituted of Scientific andTechnological

Research,Thailand)was used for koji fermentation.

Saccharomyces cerevisiaeSC90(kindlyprovidedby

AssociateProfessorDr.TawornVinijsanun,Mahidol

University, Kanchanaburi Campus, Thailand)was

usedduringalcoholicfermentation.Attheendofkoji

fermentation, total nitrogen (TN), total soluble solid

(TSS)ofsugaryliquidweremeasured.Duringalcoholic

fermentation, alcohol level, amino acid content and

concentrationofvolatilecompoundsweredetermined.

Allexperimentswerecarriedoutintriplicate.

2.2 Materials

Glutinous ricewas purchased from a local

market inKanchanaburi, Thailand.Wheat ricewas

kindlyprovidedfromPiyasattree,Ltd.,Nakhonpathom,

Thailand.Theserawmaterialswerekeptinadessicator

941KKU Res. J. 2012; 17(6)

atroomtemperaturethroughoutexperiment.

2.3 Production of Sato

Glutinous rice (Oryza satava var glutinosa)

andwheatrice(Triticum aestivumL.)weremixedwith

waterat1:2and1:3forglutinousriceandwheatrice,

respectively.Theseratioswereusedtoensureoptimum

kojifermentation(datanotshown).Theyweresoaked

overnightpriortosubjecttoheatat121oCfor15min.

TenmillilitersofA. nigerTISTR3257sporesuspension

(107spores/mL)weremixedwiththecooledrice(600g).

Thekojifermentationwascarriedoutat30oCfor7days.

Afterthat,theTSSofkojiwasadjustedto22oBrix.Free

sulfurdioxidewasadjustedto200ppmusingsodium

metabisulphite.Ammonium sulphatewas added into

thesampleto1g/Lconcentration.Then,S. cerevisiae,

previouslygrownovernight(107CFU/mL)inpineapple

juicesupplementedwith1g/Lofammoniumsulphate,

was inoculated at 4% (v/v). The fermentationwas

carriedoutin1LErlenmeyerflaskunderstaticcondition

at30oCfor8days.

2.4 Analysis during koji and alcoholic

fermentation

Glutinousriceandwheatricewereanalyzed

fortheircompositionsincludingmoisture,protein,total

nitrogen,fatandashcontentaccordingtoAOAC(10).

Total soluble solid was measured using a hand

refractometer.Alcoholcontentofsampleswasexamined

usinganEbuliometer.Freeaminoacidcontentsincluding

leucine, isoleucine, phenylalanine and valinewere

determined by using high performance liquid

chromatography (HPLC 200 series, Perkin elmer,

U.S.A.)connectedwithdiodearraydetector(338nm)

accordingtomethoddescribedbySouflerosandothers

(11).ThecolumnusedwasaUPSC18,5 mcolumn,4.6 x 250mm,VerticalChromatography,Thailand).

Volatile compounds including 3-methyl-1-butanol,

2-methyl-1-propanol,phenylethanol,isobutylacetate,

isopentylacetate,phenetylacetateanddiethylsuccinate

were determinedby using gas chromatography-mass

spectrophotometry(GC-MS7890A,AgilentTechnologies,

U.S.A.)couplingwitha007-CWcapillarycolumn(30m

x0.25mmdiameter,0.25mmfilmthickness(Quadrex,

U.S.A.).Liquid-liquidextractionandrunningcondition

were carried out according tomethod described by

Ortega,López,CachoandFerreira(12).

2.5 Statistical analysis

Theexperimentalresultswereanalyzedusing

Student’sttestat95%confidence.

3. Results and Discussion

3.1 Composition of glutinous and wheat rice

Compositions ofGlutinousRice (GR) and

Wheat Rice (WR) are shown in Table 1. TheWR

sample contains significantly higher protein content

(~6.6),totalnitrogen(~1.2),fatcontent(~0.9)andash

content(~1.4)thantheGRsample(p<0.05).Incontrast,

theGRsamplehassignificantlyhighermoisturecontent

(~3% higher) and total carbohydrate (~9% higher)

comparedwiththeWRsample.

942 KKU Res. J. 2012; 17(6)

Table 1.CompositionsofGlutinousriceandWheatrice(%,drybasis)

Composition (%) Glutinous rice Wheat rice

Moisturecontent 13.17±0.05a 10.60±0.07b

Proteincontent 6.98±0.07b 13.53±0.06a

Totalnitrogen 1.17±0.01b 2.37±0.01a

Fatcontent 1.65±0.07b 2.59±0.08a

Ashcontent 0.31±0.02b 1.70±0.01a

Totalcarbohydrate* 91.06±0.08a 82.18±0.13b

a,bMeanvaluesshowingdifferentsuperscriptletterdiffersignificantly(p<0.05)

*Totalcarbohydrate=100–(Protein–Totalnitrogen–Fat–Ashcontent)

3.2 Total soluble solid (oBrix) and total nitrogen (%,

dry basis) of koji

Attheendofkojifermentation,bothsamples

(GRandWRsample)wereanalyzedfortotalsoluble

solid (Figure 1a) and total nitrogen (Figure 1b).The

totalsolublesolidintheGRsampleishigherthaninthe

samplemadefromwheat,21.53vs.17.13oBrix,respectively.

Ontheotherhand,theWRsamplecontainshighertotal

nitrogen content comparedwith the control sample,

0.24vs.0.085%(w/w),respectively.Itiswellknown

thatAspergillus spp.producesbothamylasesandacid

proteases(5,13,14).Theseenzymeshydrolyzesubstrates

suchasstarchandproteinintosugarsandaminoacids.

Theresultsoftotalsolublesolidandtotalnitrogenshow

similar trendsasthatfoundincompositionalanalysis

ofrawmaterial(Table1, totalcarbohydrateandtotal

nitrogen). Thismight suggest that the difference in

concentrationofsugarsandnitrogenofkoji isdueto

compositionof rawmaterials used.Nevertheless, the

findingmaybepartlyduetoinfluenceofrawmaterialson

activityofhydrolyzingenzymesproducedbythefungus.

Figure 1.Totalsolublesolid(oBrix)andtotalnitrogen

(%,drybasis)ofkojiattheendoffermentation(7days)

3.3 Total soluble solid, total nitrogen and

alcohol content during alcohol fermentation

Theresultsoftotalsolublesolid,totalnitrogen

andalcoholcontentduringalcoholicfermentationare

presentedinFigure2.Atthebeginningoffermentation,

thesugarlevelofbothsampleswasadjustedto22oBrix

(Figure 2a).The sugar level decreased rapidly in the

943KKU Res. J. 2012; 17(6)

samplemadeusingwheat riceduring thefirst2days

from22to10oBrix.Ontheotherhand,inthecaseofthe

controlsample,thesugarlevelslowlydeclinedfrom22

to17oBrix.Thereafter,theutilizationofsugarreached

aplateauatday6fortheGRsampleandday4forthe

WRsample,respectively.Patternofnitrogenreduction

issimilar to thatofsugarutilizationwhere therewas

higher reductionof total nitrogenof theWR sample

comparedwiththecontrolsampleduringthefirst2days,

anditcontinuedtodeclineuntiltheendoffermentation

(Figure2b).However,inthecaseoftotalnitrogen,the

initialconcentrationintheWRsample(2.8%)washigher

thanthatofthecontrolsample(1.4%)(Figure2b).By

takingthevalueoftotalnitrogenintoconsideration,the

increaseinsugarutilizationintheWRsampleislikely

tobedue to itsnitrogencontent.Similar justification

shouldalsobesuitableforexplainingtheresultofalcohol

content(Figure2c).Thealcoholcontentofbothsamples

rosesteeplyduringthefirst2dayswiththeWRsample

showinghigher rate of alcohol production compared

with thecontrolsample.Then, thealcoholcontentof

both samples slowly increasedwith comparable rate

untiltheendoffermentation.Attheendoffermentation,

theGRandWRsamplescontain10.8and12.7%(v/v),

respectively. Similar results about the effect of total

nitrogen (both free amino acids and ammonium) on

sugar utilization and alcohol production have been

reportedbyHernández-Orte,Ibarz,CachoandFerreira

(5),Arias-Gil,Garde-CerdánandAncín-Apilicueta(15)

andTaillandier,Portugal,FusterandStrehaiano(16).It

hasbeenshownthatyeastconsumedaminoacidsmainly

inthefirstquarterofalcoholfermentation(5).Inaddition,

therateofaminoacidconsumptionispositivelycorrelated

with its initial concentration (16). However, their

experimentdidnotfoundacorrelationbetweenrateof

sugarassimilationandconcentrationofnitrogen.

Figure 2.Totalsolublesolid(oBrix),totalnitrogen(%,

wetbasis)(b)andalcoholcontent(c)duringalcoholic

fermentation

3.4 Concentration of valine, phenylalanine,

leucine and isoleucine during alcohol fermentation

Concentrationoffreeaminoacidsincluding

valine,phenylalanine,leucineandisoleucinemeasured

duringalcoholfermentationoftheGRandWRsamples

isshowninFigure3.Theconcentrationofallaminoacids

studiedintheGRsample(10-80mg/L)waslowerthan

thatoftheWRsample(15-130mg/L).Thismightbedue

tothedifferenceoftheirinitialnitrogenconcentration

(Figure1b).Moreover,intheGRsample,therewasan

increasingtrendinconcentrationofallfouraminoacids

944 KKU Res. J. 2012; 17(6)

fromthebeginningtotheendoffermentation(Figure

3a).On the contrary, in theWRsample (Figure 3b),

onlyconcentrationofvalineand isoleucine increased

to theirmaximum at day 4 and 6, respectively, and

declinedafterward.Thisisnotthecaseforphenylalanine

andleucine.Theirconcentrationfirstlydeclinedduring

thefirst2daysfollowedbyasteeply increased to its

maximumatday4(phenylalanine)andday6(leucine).

Then,theirconcentrationdecreasedagainfromday6

today8.

Figure 3.Concentrationof free amino acids (valine,

phenylalanine, leucine and isoleucine) in the sample

madewithglutinousrice(a)andwheatrice(b)measured

2daysintervalduringalcoholfermentation

Ascanbeseen,theresultsoffreeaminoacid

content (Figure 3) contradicts to the change of total

nitrogen (Figure 2b).Generally, the amino acids and

peptidesareusedbyyeast(17).However,someamino

acids,suchasisoleucineandleucine,canbeproducedby

yeastduringwinefermentation(15).Iemura,Yamada,

Takahashi, Furukawa andHara (18) observed that

whentheinitialaminoacidsinmediumarehigh,yeast

preferstoassimilateaminoacidsfromthemediumover

peptides.Thisdecreasesfinalaminoacidconcentration.

Barrajón-Simancas,Giese,Arévalo-VillenaandBriones

(17)reportedsimilarresultthattheconcentrationofmost

freeaminoacidsinwinedecreasesfromthebeginning

to the endof fermentation.Thesefindingsoppose to

ourresult that theconcentrationofsomeaminoacids

increasedcontinuouslyduringalcoholfermentation.This

discrepancymightbeduetothefactthattheproteolysis

ofriceproteinbyenzymeproducedfromkojioccurred

duringfermentationasobservedbyNegiandBanerjee

(13,14)andHashizumeet al.(8),whiletheproteolysis

of grapeprotein is limitedduringwine fermentation.

Althoughthereisnoenoughevidencefromourexperiment

aboutthecauseofsuchresults,itispossiblethatthereis

higherrateoffreeaminoacidproduction(proteolysisby

acidproteasesorproductionbyyeast)thanrateofamino

acidutilization(yeastmetabolism).Highconcentration

offreeaminoacidsintheWRsamplecouldbedueto

effectsofhighnitrogencontentofwheatrice(Table1.).

However,thedifferenceinproteaseactivitybetweenthe

GRandWRkojishouldalsobetakenintoconsideration.

3.5 Monitor of volatile compounds

Production of some volatile compounds

commonly detected in alcoholic beverages including

3-methyl-1-butanol, 2-methyl-1-propanol, phenyl

ethanol, isobutyl acetate, isopentyl acetate, phenetyl

acetate and diethyl succinateweremonitored during

fermentation (Figure 4).These compounds represent

pleasantandunpleasantodordescriptionasdescribed

inTable 2.The concentrationof 3-methyl-1-butanol,

phenylethanol,phenetylacetateanddiethylsuccinate

washigherintheWRsamplecomparedwiththeGR

sample. The first two compounds increased steeply

during the first 2 days from 20 to 110mg/L for

945KKU Res. J. 2012; 17(6)

3-methyl-1-butanolandfrom36to69mg/Lforphenyl

ethanolinthesample,respectively.Theirconcentration

rosetothemaximum(122mg/Lfor3-methyl-1-butanol

and89mg/L forphenyl ethanol) at day6.Then, the

concentrationofphenylethanoldeclineddramatically.

Theconcentrationoftheothertwocompounds(phenetyl

acetate and diethyl succinate) just slightly increased

duringfermentation.Theirconcentrationwerebetween

6.0 to 7.0mg/L (GR sample), 7.0 to 8.0mg/L (WR

sample)forphenetylacetateand0.2to0.4mg/L(GR

sample), 0.4 to 0.5mg/L (WR sample) for diethyl

succinate. For isopentyl acetate and isobutyl acetate,

theirconcentrationswererelativelyconstantinthecase

ofWRsample.However,therewashigherfluctuationin

concentrationfortheGRsample.Theconcentrationsof

bothcompoundswerebetween2.0to3.5mg/L(isopentyl

acetate)and1.0to3.0mg/L(isobutylacetate)inboth

samples.TheGRsamplecontainedhigherconcentration

of2-methyl-1-propanolcomparedwiththeWRsample

throughoutalcoholfermentation.Theirconcentrations

werebetween18to50mg/LfortheGRsampleand15

to35mg/LfortheWRsample.

In general, these compounds belong to two

groups,whicharehigheralcohol(3-methyl-1-butanol,

2-methyl-1-propanol,phenylethanol)andester(isobutyl

acetate, isopentyl acetate, phenetyl acetate, diethyl

succinate).Estersgivefloralandfruityaromatoalcoholic

beveragessuchaswine,beerandsake(4).Higheralcohols

atasuitableconcentration(less than300mg/L)have

beenreportedtogivecomplexitytowine(19,20).When

theconcentrationsofthemarehigherthan400mg/L,they

giveanunpleasantaroma.Valine,phenylalanine,leucine

andisoleucineareprecursorsof2-methyl-1-propanol,

phenyl ethanol, 3-methyl-1-butanol, and 2-methyl-

1-butanol, respectively (3).Higher alcohols, one of

thesubstrateinvolvedinestersformation,areformed

fromaminoacidsavailableinfermentingmediumby

Ehrlich pathway (20).Amino acids are converted to

a-ketoacidfollowedbyconversiontohigheralcohols.

Acetateesters(phenethylacetate,isopentylacetate)are

formed from the reaction of acetyl-CoAandhigher

alcohols.Ascanbeseenfromtheresultsoffreeamino

acids(Figure3),higherconcentrationofhigheralcohol

(3-methyl-1-butanol and phenyl ethanol) in theWR

samplecouldbeduetothehighconcentrationofleucine

andphenylalanine.Theincreaseofphenylethanoland

phenylacetateinthisstudyissimilartotheresultsof

Hernández-Orte, Ibarz,CachoandFerreira (5)which

theincreasedvolatilecompoundswasassociatedwith

anincreaseinconcentrationofphenylalanine,aspartic

acid,threonineandalanine.However,highconcentration

ofvalineintheWRsampledidnotcauseanyincrease

in2-methyl-1-propanol.Inaddition,theproductionof

acetateestersdoesnotcorrelatewiththeconcentrationof

theiraminoacidprecursors.Theseresultscouldbedueto

yeaststrainsasobservedbyPatelandShibamoto(21)and

Torrea,Fraile,CardeandAncín(22).Theyobservedthat

differentstrainsofS. cerevisiaeshoweddifferentability

onproductionofvolatilecompounds.Foracetateesters,

theconcentrationsofthetwosampleswererelatively

constant.Thissuggeststhatyeastdoesnotproducethese

compoundsduringalcoholfermentationregardlessofthe

effectofaminoacidconcentration.Ourresultscontradict

tothoseofotherstudies(2,5,9,15,16).Verstrepenetal.

(4)hasreviewedthatoxygenisonecontributingfactor

forrepressionofalcoholacetyltransferases,theenzymes

responsible for esters formation. In our study, the

fermentationwas carried out in a Erlenmeyerflask

coveredwithacottonplug.It ispossiblethatoxygen

can penetrate into the flask. This would repress

alcohol acetyltransferases of yeast andprevent esters

fermentationinourstudy.

946 KKU Res. J. 2012; 17(6)

Figure 4.Volatilecompoundsincluding3-methyl-1-butanol(a),2-methyl-1-propanol(b),phenylethanol(c),

isobutylacetate(d),isopentylacetate(e),phenetylacetate(f)anddiethylsuccinate(g)ofricewinemadeusing

glutinousrice(◆)andwheatrice(■)duringalcoholfermentation.

947KKU Res. J. 2012; 17(6)

Interestingly, some of the volatiles studied

wereobservedatthefirstdayofalcoholicfermentation

suggestingthatthesecompoundsarealreadyproduced

duringkojifermentation.AccordingtothestudyofMo,

Xu andFan (23), they observed volatile compounds

producedinQu,amoldedcerealusedforChineserice

wineproduction.Thiswouldconfirmourspeculation

thatthevolatilecompoundsarealreadyproducedduring

kojifermentation.However,therewasnodifferencein

theinitialconcentrationofvolatilecompoundsamong

two samples. Thismight suggest that therewas no

effectofrawmaterial,whichishighinnitrogencontent,

onproductionofthevolatilecompoundsstudiedduring

kojifermentation.

Withregardtoodoractivity,theodoractive

value (OAV),which represents activityofvolatile in

sample,canbecalculatedbydividingtheconcentration

ofvolatilecompoundswiththeodorthresholdvalue(24).

TheOAVof3-methyl-1-butanol,2-methyl-1-propanol,

phenyl ethanol, Isobutyl acetate, Isopentyl acetate,

phenetyl acetate and diethyl succinate at the end of

fermentationare1.51,0.63,0.20,1.18,18.25,3.84and

0.19fortheGRsampleand1.88,0.33,0.24,2.19,15.19,

4.09and0.43fortheWRsample,respectively(Table

2.). This result shows that esters are potent volatile

compoundinricewine(valuemorethan1).Basedon

theOAVvaluesof3-methyl-1-butanol,isobutylacetate,

isopentylacetateandphenetylacetate,theGRandWR

sampleswould have different flavor characteristics

(fruity,sweet,bananaandflowery).Inaddition,although

there are volatile compounds that have the OAV

value lower than 1 in both samples,Ryan, Prenzler,

SalibaandScollary(25)havepointedoutthatvolatile

compoundswhichhavelowimpactodorants(OAVless

than1.0)influenceglobalsensoryperceptionofwine.

Therefore, theWR sample containing higherOAV

valuesofphenylethanolanddiethylsuccinate(giving

positiveflavorcharacteristics) thanintheGRsample

maygivebetterflavorcharacteristicovertheGRsample.

However,othervolatilesthathavenotbeendetermined

inthisstudyalsocontributetoflavorcharacteristicof

ricewine.Thoroughanalysisofvolatilecompoundsas

wellassensoryevaluationisneededtobecarriedoutto

confirmourpresumption.

Table 2.Odoractivevalue(OAV)ofvolatilecompounds

Volatile compounds Odor threshold (mg/L)* Glutinous rice Wheat rice Odor description

3-methyl-1-butanol 60.00 1.51±0.03 1.88±0.15 Solvent

2-methyl-1-propanol 75.00 0.63±0.05 0.33±0.01 Alcohol,nailpolish

phenylethanol 200.00 0.20±0.07 0.24±0.10 Rose,honey

Isobutylacetate 1.60 1.18±0.54 2.19±0.79 Fruity,apple,banana

Isopentylacetate 0.16 18.25±2.13 15.19±1.01 Banana,fruity,sweet

phenetylacetate 1.80 3.84±0.31 4.09±0.09 Flowery

diethylsuccinate 1.20 0.19±0.01 0.43±0.01 Fruity,melon

*Valuesofodorthreshold(mg/L)comefromPeinado,Moreno,Bueno,MorenoandMauricio(26)

948 KKU Res. J. 2012; 17(6)

4. Conclusion

Inconclusion,itcanbeseenthattheuseofraw

materialthatishighinnitrogencontentsuchaswheatrice

usedinthisstudyaffectskojiandalcoholfermentation

aswellassomevolatilecompounds.Theeffectoffree

amino acids as a result of proteolysis during alcohol

fermentationonwinevolatilecompoundsmainlyinfluences

higheralcohols(3-methyl-1-butanolandphenylethanol).

Ascertainvolatilecompoundscanbedetectedatthefirst

dayoffermentation,thissuggeststhattheyarealready

producedbymoldduringkojifermentation.

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