全 文 ：第 28卷第 1期
2008年 2月
林　产　化　学　与　工　业
ChemistryandIndustryofForestProducts
Vol.28 No.1
Feb.2008
ChemicalConstituentsandAntioxidantActivitiesofthe
ExtractivesfromJuglansmandshuricaMaxim.Bark
　　收稿日期:2006-12-21
　　基金项目:中国博士后科学基金(20070410198);天津科技大学引进人才科研启动基金(20060434)
　　作者简介:司传领 (1975-), 男 , 山东菏泽人 , 副教授 , 博士 , 主要从事林产化工 、天然产物方面的研究;
E-mail:sichli@yahoo.com。
SIChuan-ling　　
SIChuan-ling1, 2 , LIUZhong1 , HUILan-feng1 , KIMJinKyu2 , BAEYoungSoo2
(1.TianjinKeyLaboratoryofPulp＆Paper, TianjinUniversityofScience＆Technology, Tianjin300222,
China;2.ColegeofForestSciences, KangwonNationalUniversity, Chuncheon200-701, Korea)
Abstract:Onthebasisofchemicalandspectroscopicevidence, 11 compounds, including3 flavanols:
pinobanksin(1), taxifolin(2)andampelopsin(3), 3 flavonols:kaempferol(4), quercetin(5)and
myricetin(6), and5 flavonoidglycosides:afzelin(7), astragalin(8), quercitrin(9), isoquercitrin
(10)andmyricitrin(11), wereisolatedfromthestembarkofJuglansmandshuricaMaxim.Thiswastheisolationofcompounds
1, 3, 5, 6, 8 and10fromthisspeciesforthefirsttime.Theantioxidantactivitiesofthecrudeextract, partitionedfractionsand
isolatedcompoundswereevaluatedbyDPPHfreeradical-scavengingassay.ResultsshowedthatEtOAcandH2Opartitioned
fractionsandcompounds1-6 exhibitedstrongantioxidantactivitiescomparedwithcontrols.
Keywords:JuglansmandshuricaMaxim.;bark;antioxidantactivity
CLCnumbers:TQ91;Q949.754.7　　　　　Documentcode:A　　　　　ArticleID:0253-2417(2008)01-0029-04
核桃楸树皮提取物的化学成分及其抗氧化活性研究
司传领 1, 2 , 刘 忠 1 , 惠岚峰1 , 金辰奎 2 , 裵映寿 2
(1.天津科技大学天津市制浆造纸重点实验室 , 天津 300222;
2.韩国江原大学 森林科学学院 , 韩国 春川 200-701)
摘　要:研究了核桃楸树皮的化学成分及其抗氧化活性。采用 SephadexLH-20柱色谱及薄层色谱等方法进行分离 ,从
其 70%丙酮提取物乙酸乙酯及水溶性部分中分到 11种化合物 , 经波谱分析及理化性质鉴定化合物分别为:短叶松素
(1)、 花旗松素(2)、蛇葡萄素(3)、 山奈酚(4)、 槲皮素(5)、杨梅素(6)、 阿福豆苷 (7)、 紫云英苷 (8)、 槲皮苷(9)、
异槲皮苷(10)、杨梅苷(11)。化合物 1、 3、 5、 6、 8、 10为首次从该植物中分得。经 DPPH试验 , 测定了正己烷溶性 、二
氯甲烷溶性 、乙酸乙酯溶性和水溶性部分以及粗提物和分得化合物的抗氧化活性。其中乙酸乙酯和水溶性部分及化合
物 1 ～ 6与对照组相比具有很强的抗氧化活性。
关键词:核桃楸;树皮;抗氧化活性
JuglansmandshuricaMaxim.(Juglandaceae)isafast-growingdeciduoustreewidelydistributedin
China, SiberiaandKoreanPeninsula.Thetreehasbeenusedasafolkmedicinalplantfortreatmentof
esophageal, gastric, cardiacandlungcancer.Itisalsoreportedthatchemicalsvolatilizedfromthespecies
inhibitthegrowthoftheneighboringplantsandcanbedevelopedforchemurgy.Severalnaphthoquinones,
naphthalenylglycosides, α-tertalonylglucopyranosides, flavonoidsanddiarylheptanoylglucopyranosideshave
beenisolatedfromthisplant, andthesecompoundshavebeenshowntohaveanti-HIV-1 activity, cytotoxic
activitytohumancancercellinesandinhibitoryefectsonDNApolymeraseandRNaseHactivitiesofHIV
reversetranscriptase[ 1-6] .Inthispapertheisolationof11 flavonoidsandflavonoidglycosidesfromthestem
barkofthisplantandtheirantioxidantactivitiesarereported.
30　 林　产　化　学　与　工　业 第 28卷
1　Experimental
1.1　Plantmaterials
J.mandshuricatreesampleswereobtainedintheexperimentalforestofKangwonNationalUniversity,
KoreainApril, 2005 andthestembarkwasstrippedfromthefreshlyfalentree.
1.2　Apparatusandreagents
1Hand13CNMRspectrawererecordedinMeOH-d4 withanAvanceDPX400instrument.FAB-MSwere
obtainedinapositivemodewithamicromassautospecM363spectrometer.Meltingpointsweredeterminedon
anElectroThermal9100 apparatusandareuncorrected.OpticalrotationsweremeasuredwithaJASCODIP-
1000digitalpolarimeterinMeOH.SephadexLH-20wasusedforcolumnchromatographyand25DC-Plastikfo-
lienCeluloseF(Merk)platesforTLC, whichweredevelopedinHOAc-H2O(3∶47 , volumeratio, sameas
infolowing)andt-BuOH-HOAc-H2O(3∶1∶1).SubstancesweredetectedbyUVlightorbyspraying
vanilin-HCl-EtOH(60∶0.15∶6)folowedbyheating.
Alreagents(DaejungCo.Ltd.Korea)usedinthisworkwereofanalyticalgrade.
1.3　Experimentalprocedures
1.3.1　Extractionandfractionation　Air-driedandgroundstembark(3.2kg)wasextractedwithacetone-
H2O(7∶3).Removalofthesolventyieldedadarkbrownextract, whichwassuccessfulyfractionationedwith
n-hexane, CH2Cl2 andEtOAcinaseparatoryfunnelandfreeze-driedtogive15.6gofn-hexane, 20.6gof
CH2Cl2 , 130gofEtOAcand337.6gofH2Opartitionedfractions.
1.3.2　Isolation　AportionofEtOAcparticipatedpowder(32.0g)waschromatographedonaSephadex
LH-20 columnelutedwithMeOH-H2O(4∶1)togive5majorfractionslabeledasE1(1.06g), E2(17.1g),
E3(7.9g), E4(3.2g)andE5(2.3g), respectively.FractionE3 wasreappliedonacolumnforfurtherpuri-
ficationwithMeOH-H2O(2∶1)afording3 subfractionsasE31-E33.E31 waselucidatedascompound10
(15mg).SubfractionE32 wasretreatedonacolumnforfurtherpurificationwithMeOH-H2O(1∶2 and1∶5)
andEtOH-hexane(3∶2 and1∶2), afordingcompounds1 (20mg), 2(64mg), 3(84mg)and8(64mg).
FractionE1 wasrechromatographedonacolumnwithMeOH-H2O(2∶1 and1∶3)togivecompounds6
(12mg)and7 (40mg).
AportionoftheresultingH2Oparticipatedpowder(31.5g)wasalsosubjectedtoaSephadexLH-20
columnwithMeOH-H2O(1∶1)aselutingsolventtogive4 mainfractionsasW1 -W4.FractionW1 wasalso
appliedonacolumnwithMeOH-H2O(1∶3 , 1∶5and3∶1)toisolatecompounds4 (26mg), 5(30mg), 9
(22mg)and11 (18mg).
1.3.3　DPPHfreeradicalscavengingassay　Theantioxidantactivitywasdeterminedonthebasisofthe
scavengingactivityofthestableDPPHfreeradicalmethodintroducedbyYoshida, etal[ 7] withslightmodifica-
tion.MeOHsolutions(4mL)ofsamplesatdiferentmassconcentrations(2-40mg/L)wereaddedtoasolu-
tionofDPPH(1.5×10-4 mol/L, 1mL)inMeOH.Aftermixinggentlyandstandingatroomtemperaturefor
30min, theopticaldensitywasmeasuredat517nmwithaUV-visiblespectrophotometer(LibraS32, Bio-
chromLTD).Theresultswerecalculatedbytakingthemeanofaltriplicatedvalues.IC50 valueswere
obtainedthroughextrapolationfromconcentrationofsamplenecessarytoscavenge50% oftheDPPHfree
radicals.BHTandα-tocopherolwereusedascontrols.
2　Resultsanddiscussion
2.1　Identificationofisolatedcompounds
Threeflavanols:pinobanksin(1)20mg, taxifolin(2)64mgandampelopsin(3)84mg, 3 flavonols:
第 1期 司传领 ,等:核桃楸树皮的化学成分及其抗氧化活性研究 31　
kaempferol(4)26mg, quercetin(5)30mgandmyricetin(6)12mg, and5 flavonolglycosides:afzelin
(7)40mg, astragalin(8)64mg, quercitrin(9)12mg, isoquercitrin(10)15mgandmyricitrin(11)
18mg, asshowninFig.1wereisolatedasyelowishpowderswhichshowedphysicalandspectraldatavirtualy
identicaltothosewhichhadbeenreportedbefore[ 8-10] .
2.2　Antioxidantactivity
Theantioxidantactivitiesofthecrudeextract, participatedfractionsand11 isolatedcompoundswere
evaluatedbyDDPHassay.ResultsweresummarizedinTable1.Amongthem, theEtOAcandH2Oparticipa-
tedfractions, pinobanksin, taxifolin, ampelopsin, kaempferol, quercetinandmyricetinexhibitedpotent
Fig.1　StructuresofisolatedcompoundsfromJ.
mandshuricastembark
antioxidantactivitiescomparabletoα-tocopherolandBHT,
whichwereusedascontrols, whiletherestcompounds, par-
ticipatedfractionsandcrudeextractshowednegligibleactivi-
ties.TheresultsindicatedthattheextractivesofJ.mand-
shuricahadhighantioxidantpotentialandcanbeusedasa
usefulsourceforantioxidants.
Table1　Antioxidantactivities(IC50 values)ofthecrude
extract, participatedfractionsandisolated
compoundsfromJ.mandshurica
items samples IC50 /(mg·L-1)
controls α-tocopherol 12BHT 14
partitionedfractions
crudeextract 22
n-hexaneparticipatedfraction 43
CH2Cl2 participatedfraction 49
EtOAcparticipatedfraction 10
H2Oparticipatedfraction 12
isolatedcompounds
pinobanksin(1) 　 8.7
taxifolin(2) 8.3
ampelopsin(3) 7.8
kaempferol(4) 8.8
quercetin(5) 8.5
myricetin(6) 8.0
afzelin(7) 64
astragalin(8) 35
quercitrin(9) 16
isoquercitrin(10) 26
myricitrin(11) 33
　　Ingeneral, theflavonoidswithfreehydroxylgroupsactedasfreeradical-scavengers, andthehydroxyl
groupontheB-ringenhancedtheirantioxidantactivities:Forflavanols, ampelopsin(7.8mg/L)>taxifolin
(8.3mg/L)>pinobanksin(8.7mg/L);Forflavonols, myricetin(8.0mg/L)>quercetin(8.5mg/L)>
kaempferol(8.8mg/L).
Flavanolsshowedhigherantioxidantactivitiesthantheircorrespondingflavonols:ampelopsin(7.8mg/L)>
myricetin(8.0mg/L), taxifolin(8.3mg/L)>quercetin(8.5mg/L), andpinobanksin(8.7mg/L)>
kaempferol(8.8mg/L).
Kaempferol(8.8mg/L), quercetin(8.5mg/L)andmyricetin(8.0mg/L)showedcomparatively
higherantioxidatntactivitiesthanafzelin(64mg/L)andastragalin(35mg/L), quercitrin(16mg/L)and
isoquercitrin(26mg/L), andmyricitrin(33mg/L), respectively, indicatingthatthefreeradicalscavenging
activitiesofglycosidicderivativesweresignificantlyreducedcomparedwiththeiroriginalflavonoidaglycones.
32　 林　产　化　学　与　工　业 第 28卷
3　Conclusion
Threeflavanols:pinobanksin(1), taxifolin(2)andampelopsin(3), 3 flavonols:kaempferol(4),
quercetin(5)andmyricetin(6), and5 flavonolglycosides:afzelin(7), astragalin(8), quercitrin(9),
isoquercitrin(10)andmyricitrin(11), wereisolatedfromtheJ.mandshuricaM.stembarkbycolumn
chromatographyusingSephadexLH-20 andthechemicalstructureswereelucidatedbyspectroscopyand
chemicalevidence.Thoughcompounds2, 4, 7, 9 and11 havepreviouslybeenreportedfromthistree[ 11] , to
ourknowledge, thiswasthefirst-timeisolationofcompounds1, 3, 5, 6, 8and10 fromthisspecies.
Theresultsindicatedthatisolatedcompoundscontainingmanyhydroxylgroupsexhibitedbeterantioxi-
dantactivities.Theantioxidantactivitiesoftheflavanolswerestrongerthantheircorespondingflavonols, and
thoseofflavonoidglycosidesweredecreasedcomparedwiththeircorrespondingaglycones.
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(上接 22页)
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