Identification of new citrus norisoprenoids in orange juice using time intensity GC-0 and GC-MS

2004·3.4 MB·English

by  Mahattanatawee, Kanjana

IDENTIFICATIONOFNEWCITRUSNORISOPRENOIDSINORANGEJUICE
USINGTIMEINTENSITYGC-OANDGC-MS
By
KANJANAMAHATTANATAWEE
ADISSERTATIONPRESENTEDTOTHEGRADUATESCHOOL
OFTHEUNIVERSITYOFFLORIDAINPARTIALFULFILLMENT
OFTHEREQUIREMENTSFORTHEDEGREEOF
DOCTOROFPHILOSOPHY
UNIVERSITYOFFLORIDA
2004
Copyright2004
by
KanjanaMahattanatawee
Thisdissertationisdedicatedtomylatefather.
ACKNOWLEDGMENTS
IwouldespeciallyliketothanktoDr.RussellL.Rouseff,mysupervisor,forhis
friendshipandguidancethroughoutmygraduateprogram. Heprovidedmewith
intellectual,thoughtfuldiscussion;encouragement;andtime. Withouthishelpand
supportIwouldnothavecomefromThailandandcontinuedattheUniversityofFlorida.
Hishighethicalstandardsandphilosophicalviewswillneverbeforgotten. Iwouldlike
tothankDr.R.M.Goodrichforallofhersupport,guidanceandfriendship. Shewasvery
kindandhelpfulwheneverIneededherhelp. Ialsothanktheothermembersofmy
supervisorycommittee.Dr.D.H.PowellandDr.M.R.Marshall,Jr,fortheirkindnessand
valuable,thoughtfuldiscussiontowardmyresearch. EspeciallywarmthanksgotoDr.
KevinL.Goodnerforhisadviceandsupportoninstrumental. Myspecialthanksgoto
JohnSmootandDr.FilominaValimwhoassistedmeanytimeIneededthem. My
appreciationgoestomydearfriendDr.FahiemEl-BoraiKorawhoassistedmewhenever
Ineededhishelp. IextendmyappreciationtoallmyfriendsatCitrusResearchand
EducationCenter(CREC)inLakeAlfredfortheirfriendshipandsupport. Iwouldlike
alsotothankmyprofessorsbackinThailandexpeciallyDr.TweeHormchong,who
taughtmewhatgoodscientistsandteachersare. Mygreatthanksgotomybeloved
family. Mylovelywonderfulmother,father,brothersandsistersalwaysencouragedme
tofollowmydream,andwithouttheirloveandsupport,IcouldneverbewhoIamtoday.
Finally,myappreciationgoestotheSiamUniversityforgivingmethefinancialsupport
necessarytoobtainmyPh.DfromtheUniversityofFlorida.
iv
TABLEOFCONTENTS
cage
ACKNOWLEDGMENTS iv
LISTOFTABLES viii
LISTOFFIGURES ix
ABSTRACT xii
CHAPTER
1 INTRODUCTION 1
2 LITERATUREREVIEW 3
OrangeJuiceAroma 3
Carotenoids 3
Norisoprenoids 4
NorisoprenoidsFormationfromCarotenoids 5
Apple 7
Tomato 8
Saffron 9
GrapeandWine 10
GasChromatography-Olfactometry U
SolidPhaseMicroextraction 13
OrangeJuiceNorisoprenoids 14
3 HPLCDETERMINATIONOFCAROTENOIDNORISOPRENOID
PRECURSORSINORANGEJUICE 15
Introduction 15
Objectives 17
MaterialsandMethods 17
CarotenoidExtraction 17
CarotenoidSaponification 17
HPLCProcedure 18
ResultsandDiscussion 19
CarotenoidsofInterest 19
HydrolysisConditions 20
V
HPLCSeparation 21
CarotenoidIdentification 23
Conclusions 25
4 IDENTIFICATIONOFNORISOPRENOIDSINORANGEJUICEUSING
TIMEINTENSITYGC-0ANDGC-MS 26
Introduction 26
Objectives 28
MaterialsandMethods 28
OrangeJuiceSamplesandProcessing 28
Chemicals 29
OrangeJuiceHeadspaceExtraction 29
GasChromatography:GC-FIDandGC-OIfactometer 30
GasChromatography-MassSpectrometry 31
AromaPeakIdentification 32
ResultsandDiscussion 32
ExtractionandConcentrationofJuiceNorisoprenoids 32
GC-Olfactometry 35
MassSpectrometryNorisoprenoidIdentifications 36
Conclusion 41
5 QUANTIFICATIONANDDETERMINATIONOFTHERELATIVEIMPACT
OFNORISOPRENOIDSINORANGEJUICE 43
Introduction 43
Objectives 45
MaterialsandMethods 45
QuantificationofNorisoprenoidsinOrangeJuice 45
DeterminationoftheRelativeImpactofNorisoprenoidsinOrangeJuice 47
ResultsandDiscussion 47
QuantificationofNorisoprenoidsinOrangeJuice 47
NorisoprenoidQuantificationusingStandardAdditions 50
DeterminationofRelativeAromaImpactofNorisoprenoids 54
NorisoprenoidContributiontoTotalFloralAroma 58
Conclusion 61
6 THERMALDEGRADATIONOFBETA-CAROTENEINMODEL
SOLUTION 62
Introduction 62
Objective 63
MaterialsandMethods 63
Crystallization 63
ModelSolutions 64
AnalyticalMethods 64
Resultsanddiscussion 65
vi
GC-0AnalysisofP-CaroteneDecompositionat35°C 68
MSIdentification 70
Conclusion ^2
7 CONCLUSIONS 73
LISTOFREFERENCES 75
BIOGRAPHICALSKETCH 84
vii
LISTOFTABLES
Table ESge
3-1 HPLCretentiontimes,spectralcharacteristicsoforangejuicecarotenoids 24
4-1 Identification,retentioncharacteristicsandaromadescriptionsofaromaactive
compoundsinfreshorangejuice 36
5-1 ReproducibilityofSPMEexposuretime45minat40°C 49
5-2 Concentrationofnorisoprenoidsinfreshorangejuiceasdetermindedby
standardadditiontechnique 53
5-3 ConcentrationofP-damascenoneinfresh,pasteurizedandreconstituted
concentrate 53
5-4 Aromaactivecompoundsinorangejuicegroupedbycitrusy/minty 55
5-5 Aromaactivecompoundsinorangejuicegroupedbymetallic/mushroom/
geranium 56
5-6 Aromaactivecompoundsinorangejuicegroupedbyroasted/cooked/meaty/
spice 56
5-7 Aromaactivecompoundsinorangejuicegroupedbyfatty/soapy/green 56
5-8 Aromaactivecompoundsinorangejuicegroupedbysulfury/solventy/medicine..57
5-9 Aromaactivecompoundsinorangejuicegroupedbyfloral 57
5-10 Aromaactivecompoundsinorangejuicegroupedbysweet/fruity 57
5-11 Aromaactivecompoundsinorangejuicegroupedbygreen/grassy 58
5-12 Norisoprenoidsinorangejuiceandpeeloil 60
6-1 AromaactivecompoundsfromP-carotenethermaldegradationinmodelsolution
pH3.8,storageat35°Cfor2weeks 69
viii
LISTOFnOURES
Figure page
2-1 Examplesofcaroteneandxanthophyllcarotenoidstructures 4
2-2 Majorfragmentclassesofcarotenoidbiodegradation 5
2-3 Generalstepsfortheconversionofcarotenoidsintoflavorcompounds,showing
theformationofp-iononeandp-damascenonefromP-caroteneandneoxanthin
respectively 6
2-4 Formationofnorisoprenoidsaromacompoundsfromdifferentclassesof
precursors(i.e.,polyols,glycosides,andglucoseesters) 8
2-5 Steven'slaw,comparingtwodifferencecompounds:A=compoundA,B=
compoundB 12
3-1 PossibledegradationpathwaysfortheformationofP-cyclocitralandp-ionone
fromP-carotene 15
3-2 Carotenoidprecursorsofselectednorisoprenoidsincludingneoxanthin,the
indirectprecursorofP-damascenone 19
3-3 Chromatogramofsaponifiedcarotenoidextractfromorangejuiceseparated
usingaYMCC30reversephasecarotenoidcolumnandawater,MeOH,MTBE
ternarysolventgradient 22
3-4 AbsorbancespectraforP-carotene(a),leutoxanthin(b),andneoxanthin(c),peak
24,12and4respectively 23
4-1 GC-FID(top)andaveragetime-intensityoffourGC-0runsbytwopanelists
(inverted,bottom)offreshorangejuiceonZB-5column. Peaks5,19,21and23
correspondtonorisoprenoids,allnumbersreferstocompoundsinTable 4-1 34
4-2 Comparisonbetweentotalionchromatogramandselectedionchromatograms
(SIC)A:P-cyclocitral,B:P-ionone,C:a-ionone 37
4-3 Upper,totalioncurrentchromatogramfromorangejuiceheadspace,other
chromatogramsusingSIMatm/z190 38
IX
4-4 UpperspectrafromorangejuiceMSatRT=17.68,bottomspectraofP-
cyclocitralfromdatabaseNIST2002 40
4-5 UpperspectrafromorangejuiceMSatRT=21.94,bottomspectrafrom
standardP-iononeusingidenticaliontrapMSatidenticalretentiontime 40
4-6 UpperspectrafromorangejuiceMSatRT=20.87,bottomspectrafrom
standarda-iononeusingidenticaliontrapMSatidenticalretentiontime 41
5-1 ExposuretimebetweenSPMEfiberandtheheadspaceoforangejuicespiked
withstandardsat40°C, =P-cyclocitral, =P-damascenone,A=a-ionone,
•=P-ionone 48
5-2 StandardadditiondataforP-cyclocitralpeakareavs.addedconcentrationinfresh
orangejuice. Regressionlinecalculatedfrompeakareaatselectedmass137 50
5-3 Standardadditiondatafora-iononepeakareavs.addedconcentrationinfresh
orangejuice. Theregressionlinecreatedbypeakareaatselectedmass177vs.
a-iononeconcentration 51
5-4 StandardadditiondataforP-iononepeakareavs.addedconcentrationinfresh
orangejuice. Theregressionlinecreatedbypeakareaatselectedmass177vs.
P-iononeconcentration 52
5-5 StandardadditionP-damascenonepeakareavs.addedconcentrationinfresh
orangejuice. GC-quadrupolemassspectrometerinSIMmodeatm/z190 52
5-6 StandardadditiondataofP-damascenonepeakareavs.addedconcentrationin
reconstitutedfromconcentrateorangejuice 53
5-7 Aromagroupprofilesoffresh(),pasteurized(), andreconstitutedfrom
concentrate(•)orangejuice 58
5-8 Upperbarnorisoprenoidscontributemainlytothetotalfloralcategory,fresh=
78%,pasteurized=78%,andreconstituted=59%,lowerbarrepresentnon-
norisoprenoidsincludinglinaloolandunknown(LRI=1255) 60
6-1 ThestandardP-carotene(99%purity)asreceived(nopurification) 65
6-2 HeadspacevolatilesfromP-caroteneinmodelsolutionpH3.5at0day 66
6-3 HeadspacevolatilesfromP-caroteneinmodelsolutionpH3.5afterstorage1day
at35°C:1=p-ionone,a=sweet/raspberry 66
6-4 DegradationofP-caroteneinmodelsolutionatdifferencecarbonbonds 67
X