Koh-2014-Identification of an(Glycine max (L.) Merr.) varieties
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ORIGINAL
ARTICLE
Identificationofanthocyaninsinblacksoybean
(Glycinemax(L.)Merr.)varieties
KwangohKoh&JungEunYoun&Hee-SeonKim
Revised:2August2011/Accepted:5August2011/Publishedonline:17August2011#AssociationofFoodScientists&Technologists(India)2011
AbstractAnthocyaninpigmentsinblacksoybean(Glycine
max(L.)Merr.)varietiesasTawonkong(TW)and
Geomjeongkong-2(G2)wereidentifiedtoevaluatetheir
potentialsasnutritionalfunction,naturalcolorantorfunctional
foods.Anthocyaninextractionwasconductedwithacidified
methanolwith0.1MHCl(85:15,v/v).Identificationof
anthocyaninwasconductedbycomparisonwithpurified
standardsbyHPLCandmassspectrometry(MS).G2showed
sixdifferenttypesofpigmentsbyHPLC,whereasTW
showedsevenpigments.Threemajoranthocyanins(peaks1,
3,4)weredetectedinbothvarietiesandpeak1was
characterizedasdelphinidin-3-O-β-D-glucoside,peak3as
cyanidin-3-O-β-D-glucoside(C3G),andpeak4aspetunidin-
3-O-β-D-glucosidebycomparisonofchromatographicprop-
ertieswithauthenticstandardsandMS.Minorpeaks5,6and
7inTWweretentativelyidentifiedaspelargonidin-3-O-
glucoside,pelargonidin-3-O-(6″-malonylglucoside)andcyani-
dinonthebasisofMS.MSwithmajorionsat287and449of
peak2wereexactlysameasthoseofpeak3meaningthat
peak2hascyanidinandotherhexosedifferentfromglucose.
Afteracidhydrolysisoffractionedpeak2,HPLCshowedthe
hexoseasgalactose,andpeak2wasidentifiedasC3Glactose.Themostabundantanthocyanininblacksoybean(Glycine
max(L.)Merr.)wasC3GandG2showedthehigheramount
oftotalanthocyaninsthanTW(p<0.001).
KeywordsAnthocyanin.Blacksoybean.Natural
colorant.Massspectrometry(MS)
Introduction
Anthocyaninsaregroupsofreddishorpurpleflavonoids
thathavebeenusedwidelyasnaturalcoloringagentsinthe
foodindustry(BridleandTimberlake1997).Theyhave
beenrecognizedashealth-promotingfunctionalfood
ingredientsduetotheirantioxidantactivity(Nametal.
2006;Philpottetal.2006;Satue-Graciaetal.1997),which
hasbeenreportedtoreducetheriskofcoronaryheart
disease(StockerandO’Halloran2004;Waterhouse1995).
Anthocyaninsarealsoknowntohaveanti-cancer(Hyun
andChung2004;Kameietal.1995;Zhaoetal.2004),
hypoglycemic(Tsudaetal.2003),andanti-inflammatory
effects(Tsudaetal.2002)andhavebeenusedinthe
treatmentofvariouscirculatorydisorders(Bettinietal.
1985).Furthermore,thesefunctionsprovidesynergic
effectswithvariousnutrientsinvivo,sotheirnutritional
valuesareofgreatinterestrecently.
Consumptionsofblacksoybeansarerapidlygrowing
duetotheirnutritionalvaluesandpotentialstodevelopas
healthyfunctionalfoodingredientsinKorea.Anthocyanins,
agroupofreddishorpurpleflavonoids,arereportedtobe
theprimarypigmentsintheblacksoybeanvarieties
(Choungetal.2001;KurodaandWada1933;Leeetal.
2009;YoshikuraandHamaguchi1969).Blacksoybeans
containanthocyaninsmainlyintheseedcoat.Theblack
pigmentationisduetoaccumulationofanthocyaninsinthe
epidermispalisadelayeroftheseedcoat(ToddandVodkinK.KohDepartmentofChemistry,SoonchunhyangUniversity,646Eupnae-ri,Shinchang-myun,Asan-si,Choongnam336-745,RepublicofKorea
J.E.Youn:H.-S.Kim(*)DepartmentofFoodScienceandNutrition,SoonchunhyangUniversity,646Eupnae-ri,Shinchang-myun,Asan-si,Choongnam336-745,RepublicofKoreae-mail:hskim1@sch.ac.kr
J.E.YounKoreaFoodandDrugAdministration,Osongsaengmyeong2ro,Cheongwon-gun,Choongbuk363-951,RepublicofKoreaJFoodSciTechnol(February2014)51(2):377–381DOI10.1007/s13197-011-0493-y1993),whichcouldbeseparatedintoanthocyanin-rich
fractionsforuseasfunctionalcolorantsorfunctionalfood
ingredients.Althoughanextensivescientificliteratureon
thecompositionofanthocyaninsinfruitsandvegetables
exists(Francis2000;MazzaandGao2005),reportson
anthocyaninsindifferentvarietiesofblacksoybeanare
limitedwithcontroversialresults.Severalgroups(Choung
etal.2001;KurodaandWada1933;Yoshidaetal.1996;
YoshikuraandHamaguchi1969)haveidentifiedonlyone
tothreedifferentanthocyanins[cyanidin-3-O-β-D-glucoside
(C3G),delphinidin-3-O-β-D-glucoside(D3G)andpetunidin-
3-O-β-D-glucoside(Pt3G)],whileLeeetal.(2009)recently
reportedninedifferentanthocyanins,whichincluded
threemajoranthocyaninsandseveralotheranthocyanins.
Thesestudiesshowsubstantialdifferencesinanthocyanin
contentsandcompositionsamongvarieties.Therefore,further
investigationincludingminorpigmentsisrequiredtoproduce
ananthocyaninprofileoftheblacksoybeanfromdifferent
varieties.
Thepresentstudywasaimedatcharacterizinganthocy-
anincompositionindifferentvarietiesofblacksoybeanto
identifyananthocyanin-richvarietyforthefutureindustry
utilizationordevelopmentoffunctionalfoods.Theidentity
ofanthocyanincompoundswasbasedonthedataobtained
fromHPLCandliquidchromatography-massspectrometry
(LC-MS).
Materialsandmethods
Samplepreparation
Twodifferentvarietiesofblacksoybean,Tawonkong(TW)
andGeomjeongkong-2(G2),wereselectedforuseinthis
study.Theblacksoybeanswerecultivatedandgrownfor