天然产物研究与开发NatProdResDel,2011.23:464-472
文章编号:1001-688012011)03-0464-09
中药砂仁挥发油化学成分及其抗茵活性
张生潭1,王兆玉2,汪铁山1,李苗霞1,林敬明h
1南方医科大学珠江医院药剂科,广州510282;2广东药学院生命科学与生物制药学院,广州510006摘要:本研究将Bligh-Dyer溶剂提取法首次应用于植物挥发油的提取,通过GC-MS方法分析,从干燥成熟的砂仁种子和果壳四种挥发油提取物中共鉴定出138种化学成分。挥发油成分主要包括乙酸龙脑酯(5%一47%)、樟脑(4%-17%)、龙脑(1.5%一6%)、莰烯(O.2%一3%)、仅-蒎烯(O.2%一3%)、B・蒎烯(0.2%一5%)以及Ot.柯巴烯(O.1%一2%)等,其中总烯类物质相对含量占总挥发油总成分的10%一40%。从该植物中首次鉴定出蓝桉醇、二环大根香叶烯、薄荷烯醇、二十三烷、二十四烷等新的化学成分35个。所得挥发油对部分真菌及细菌的抑制试验表明其对红色毛癣菌、须毛癣菌、石膏样小孢子癣菌、金黄色葡萄球菌和粪肠球菌均表现出显著的抑制活性。因此,砂仁不仅可作为抗菌药物应用于临床,也可以作为食品原料或天然的食品防腐剂用于食品工业的生产。
关键词:砂仁;挥发油;CC—MS;抗菌活性;BliSh—Dyer法中图分类号:1t284.2
文献标识码:A
CompositionandAntimicrobialActivitiesofEssentialOilofFructusAmomi
ZHANGSheng.talll,WANGZhao.yu2,WANGTie—shahl,LIMiao—xial,LINJing.min91’
1Department
ofPharmacy,ZhujiangHospital,SouthernMedicalUniversity,GuangzhouSl0282,China;
2College矿舻ScienceandBiopharmacology,GuangdongPharmaceutical
Univenity,仇纰删JDD嘶,China
Abstract:’Ik
essentialoil
extractswere
respectivdyobtainedfromthes∞d8andhusksofAmomumvillosuml_ourby
u-
sing
Bligh・Dyermethodsfor
thefirsttime.Onehundredandtllirty-eightcompositions
were
identifiedbyGC・MS
from
fouressentialoilextracts.,11lemaincomponentsoftheessentialoilwerebomylacetate,camphor,bomeol,俚-pincne,13・
pinene
and
d.eopalene.etc.The
relative
contentsoftotalalkenes
and
ketonescomposition
accountedfor10%40%in
integralofessentialoil.Thirty・fivecompositionssuch
a8
neophytadiene,globulol,bicyclogermacrene,etc
were
identified
fromthis
plant铀dy.‰resultsofinhibitory
effect
forbacteriaandfun#showedthattheessentialodhadremarkable
inhibitoryeffectsforTrichophytonrubnlm,Tfichophytonmentagrophyton,Micresporumgypseum,Staphyleccecus
811reus,
andEntemcoccusfaecalls.Ther商ore.the
essential
oilCanbeusedinthefoodindustry
a8rswfoodmaterial
or
natural
foodpreservation.
Keywords:FructusAmom/;essentialoil;C,C--MS;antibacterialactivity;Bligh-Dyermethod
spleenand
antidiarrhea,etc.[21Inaddition,itisalso
a
keyflavoringinbothfood
anddrink.Theseactivities
Amom/istheripenedf,抛itsofseveralphntsofhavebeenlinked
to
thepresenceofessentialoil.L3J
Amomum(Zingiberaceae)includingA.villosum
Inthisstudy,allessentialoilwasextractedfrom
the
Lour.Vat.xanthioidesT.L.Wu
et
seedsandhusksofFructusAmom/byusingBligh・DyerandA.IongiligulareT.L.Wu.…Ithasbeen
methodandHydrodistillationmethodrespectively.Oneas
an
ingedientintraditionalchinesemedicine
hundredandthirty・eightcompositionswereanalyzedeliminatingdampness,promotingappetite,warming
andidentifiedbyGC—MSfromthefoursamples.ni啊・
fivecompositionssuch
as
neophytadiene,globulol,ger-
macrene
Received
August
26.2010;AcceptedDecember31。2010
D,fluorenone,bicyclogermacrene,piperitol,
FoundationItem:This
pmjcctW85鲫pponedbyGuant弘houMunicipal
tricosane。tetracosane。etc
were
firstidentified
inthis
ScienceandTechnologySupportProgramme(2008Zl—FA61)
toconventional・Corresponding.author
Tel:86-20-61643557;E—mllil:linjml231@210n.c锄
plant.Comparedhydrodistillationmeth-
Introduction
Fructusgenus
Lour,A.villosum
Senjen
used
for
VoL23
ZHANGSheng—tan。eta/:CompositionandAnfimicrobialActivitiesofEssentialOnofFructusAmom/465
od.21kindsofnewchemicalcompositionswereidenti.
fled.AndtheextractionefficiencyofBligh—Dyermethod
ismuch
higherthanthatofhydrodistillation
method.
ne
resultsofpharmacologicalactivityshowedthatthe
essentialoilofFructusAmomihad
significant
inhibitory
effectsforchosenbacteriaandfungi.Inthispaper,a
new
applicationforFructusAmomiisprovidedinanti—
biotics
and
foodpreservation,whilerelated
scientifictheoryandexperimentalevidenceare
alsopresentedforutilizingessentialoilofFructusAmomiinthephanna.ceuticalindustryandfoodindustryofdevelopmentandutilizatio玎.
Materials
and
Methods
General
GC-MSsystem(GC6890seriesII;MSD5973,Hew.
1ett
Packard,America),RotaryEvaporator(R-200,
BUCHI
company,Switzerland),ultrasoniccleaning
e・
quipment(KQ-600DE,KunshanUltrasonicequipment
company,China),electronicscales(BL-2000S,setracompany,America),Incubator(DNP-9162,Ningbo
SoutheastInstrumentCompany,China),biologicalsafe—
ty
cabinet(BHC.1300IIA/B3,Suzhou
Antai
Air’rech
Co,Ltd,China);Antimicrobialsusceptibility
test
discs(5×50discs,OxoidLtd,England),Medium(OxoidLtd,England).PlantMaterials
Ourresearchwascarried
out
on
FructusAmom/,inclu—
dingseeds
and
husksofdryingandripeningfruits.The
FructusAmomiwaspurchased
in
Guangzhou
herbal
company.Theoriginalpurchasingplacewas
Guang
dong
province,China(.Batch
number:
Materials
rrichophytonrubrumCMCC(F)TIa,Trichophyton
mell—
CMCC(F)T5a,andMicrosporum
gypseum
were
purchasedinSkinResearchInsti.
ofChineseAcademyofMedicalSciences.Esche・
coliATCC35218,StaphyloccocusaureR¥Rosen-ATCC29213,Enterococcusfaeeal括ATCC29212
Enterobacter
cloacaeATCC700323wer@providedMedicalTestingCenter,Zhujiang
hospital.
andanalysisoftheessentialoil
HydrodistillationmethodSeeds
andhusks
ofFructus
Amomiwerepeeledand
crushedseparately,with24meshscreen.Onehundredgramsof
plant
materialsweredistilledwith
water(1
L)fh
6hin
an
extractionequipmentinaccordance
withthedescriptionofthePharmacopoeiaofthePeo-pie’sRepublicof
China(2005)E4J.弧e
essentialoil
was
collectedanddried
over
anhydrousNa2S04,then
storedinsealedvialsat4℃.
居2趣出一助r删肠d
卟e
plant
materials(100g)and500mLofextracting
solvent(the
volumeratioofmethanol:chloroform:wa.
ter
was1:l:0.5)wereplacedina
Erlenmayerflask
(1500mL)andsoakedforthreedays.Andthenthe
flaskwastreatedthreetimeswith
ultrasonic(40kHz)
at
40±l℃f.or45min..11le
extracts
werecentrifuged
and
thesupernatantswerecombined.After
evaporation
ofthesolvent,theessentialoil
wascollectedandstored
砒4℃.
Gaschromatography-massspectrometryanalysis
弧e
essentialoilextractswereanalyzedbygas・chroma-
tographieally.’IheC,Coperatingconditionswere
as
fol-
lows:columntemperaturewasincreasedfrom50to250
℃at
arateof
5'U/min;injectortemperaturewas
set
to
250℃.andthedetector
temperaturewassetto280
℃:thecarriergas
was
He
at
a
rateof1.0mL/min.
’11leMSoperatingconditionswere船follows:carriergas
was
Heat
arate
of1.0mL/min;massspectrometryi-
onization
energy
was70
eV,and
electron
multiplier
tubevoltagewas
1905V;theionsourcetemperature
was
setto
230℃.andthequadrupoletemperature
was
setto
150℃ithemass
scan
range
was30-550amu.
’nleethylacetatesolutionoftheessentialoilwasdis-
solvedwith
1曲EtOAcandinjectedinsplitmode.
711lecompositionsinessentialoilwere
identifiedbyGC-MSbymassfragmentationpattern
and
spectralcompar-isonwithstandardsintheWiley275and
Nist05librar-
ies.
Determinationofantibacterialactivity
Invitroantibacterialactivitiesoftheessentialoil
ex.
wereevaluatedbydiscdiffusionmethodusing
Mueller-HintonAgarforbacteriaandSabouraud
Agar
for
fungi
with
determinationof
inhibition
zones
medicine20090601).
Microbiologicaltagrophyton
CMCC(F)M2a
tute
richiatracts
bachandbyPreparation
NatProdResD朗
VoL23
(Iz).‘51A100trLsuspensionoftestedbacteriaand
onon
organismsaftermacroscopicevaluation,wasdetermined
fungi(108CFU/mL)spreadedAgarmediums.106
innl
嬲MIC.whichwasexpressedinms/mL.DeterminationofMinimumBactericidaltration(M吣C)
TheMBCofessentialoilweredeterminedaccordingtotheMICvalues.Ofeachwellshowingcompleteabsenceof
Concen.
止of
and
essentialoilwasdropped
on
filterpapem
place
theAgarsurface.Standard
as
antibioticand
blackcontroldiskswereused
reference.Plateswere
incubatedat28℃for96hforfungiandat37℃for24hforbacteria,respectively.Theinhibiton
zones
were
growth,20灿was
transferred
to
agarplatesandin—24hat37℃for
es-
measuredi/i
diameters.The
nLrn
scaleofmeasurementwas
was
cubated72hat28oCforfungi
and
thefollowing:>15lO-15mm
zone
zone
stronglyinhibitory,
toni
bacteria.respectively.’11lelowestconcentrationofsentialoilwhich
no
moderately
inhibitory.and<10
viablebacteria
or
fungi
wereidenri—
,
zoneweaklyinhibitory.
fledwastheMBC.
of船nhn啪In蜥呻Co眦嘶昀。De协粤曲ption(MIC)
The
MICweremeasuredbymicrodilurionRPMI一1640
assay
recommended
by
NCCLS
susceptibility
ResuIts锄d……一一………conclusio璐
Chemicalcompositionsoftheessentialoilextracts
Onehundredandthiay・・eightcompositionswereidenri・-fledfromthefouressentialoilsamples(Table1).Theyieldsofessentialoilfromseedsdistillationmethodwere
(2000).161%e
wellconmined
96wellmicroplatewasused.each
bacteria
RPMI-1640
liquid
10止of
test
and
husksbyhydro.
medium(bacteriamL)and
concentrationof0.5-5×105CFU/
essentialoilsolution.10斗L
3.57%(g/g)and0.97%
thosebyBligh—Oyermethod
90止of
(s/g),respectively,while
were
sterilenormalsalinewasblackcontr01.Anappropriate
7.01%(s/g)and5.37%(s/g),respectively.
maincompositionsofessentialoilwerebomyl
ace.
amount
ofTween-80wasaddedtomakeitinto
a
5%
The
solution。ThisconcentrationofTween.80didnotaffectthegrowthofbacteria
rate,camphor,borneol,ot—pinene,伊pinene,and
other
and
fungi.11】efinalconcentra—components.%e
and
69.85%in
relativecontentsofcompositionsiden-
fionsoftheessentialoilinwellsrangedfrom0.00977rifledwereaccountedfor93.39%。92.18%。74.85%,
fouressentialoils,respectively,and
compositionswere
TIC
ofoil
ms/mL
were
to
72h
ms/mL.’rheincubationconditions
at28℃forfungiand24hat37℃forbac-
10.000
thoseoftotalalkenesandketonesaccountedfor10%-40%(TablesamplesshowninFigure1—4.
teria,respectively,andeachtreatmentwasduplicate
performed
in
2).The
thrice.ne
lowestconcentrationofthetest
samples,whichdidnotshow
Table1
anyvisualgrowthoftested
ChemicalconstituentsoftheessentialoilextractsofFructusAmomi
Ethylbenzenep-Xylcne
4.9635.1596.3206.3686.4536.5817.0197.1047.S78
0.020.221.48O.02
一
O.19O.66
一
Tricydo[2
21
0(2,6)]heptane,1,7,7-trimethyl-
O.cr7
●
Bicycio[310】hexane,4-methyl・1・(1-methylethyl)・
a-Thujene
a・Pina坼CampheneVerbeneneSabinene
0.7l
0.402.26O.270.090.24
2.0r70.2lO.2l
一
0.462.43
●
一
0.∞
、roL23
ZHANGSheng-tan。da/:CompositionandAnfimierobialActivitiesofEssentialOilofFru斌uaAmom/467
10口-Pine舱
7.73011Bicyclo[3
1
1】heptane,6,6-dimethyl-2-methylene-
7.73212口・Myrcene7.985131.Phellandrene8.49614a-Terpinene
8.78615Benzene,l-methyl-2・(1-methylethyl)
9.024161-1sopropyl-4-methylbenzene
9.03017Limonend
9.12518卢-Thujene
9.200191-Terplnene9.95320Linalooloxide
10.32921ds爷-Terpineol
10.37622n薯n8-Sahinenehydrate
10.38123Terpinolene
10.71924口-Campholenealdehyde
10.85525Benzene。1-methyl-4-(1・methylethenyl)・
10.91426Linalool
11.Z驺
27ci8・Sabinenehydrate
11.”7
281.6-OetMien-3-oi,3。7--climethyl-
11.28529Bicyclo[3
1
1]heptan-2-one,6,6-dimethyl
11.325301,5,7-Octauien-3-oi,3,7-dimethyl-
11.34731a-Thujone
11.70232D-Fenchylalcohol
11.88033(4-)-Nopinone
12.34334Bicyelo[3
1
1]heptan-3..ol,6,6-dimethyl-2-methylene-
12.45735hans.Pinocarveol
12.55636Camphor12.67037a.Phellandren-8-01.
12.74638a-Thujone12.89439
2(i0)-Pinen-3-one
12.979柏lsoborneol
13.12041Camphol
13.405423-Cyclohexen-I-ol。4-methyl-1-(1一methylethyl)-
13.57143
4-Terpinenol
13.641
44Bieyclo[3ll】hept-2-ene-2-carboxa
ldehyde,6,6-dimethyl・
13.92845Myrtenal
13.96746Bieyclo[3
l
l】hept-2-ene-2一methanol,6,6-dimethyl・
14.02847Myrtenol
14.10448Bicyclo[3ll】hept-3-en-2-one,4,6,6-trimethyl・。(1s)・
14.28349Verben帆e
14.3295011'1m8・Piperltol
14.41251
n哪hyl
acetate
14.490
O.2l
0.630.06O.10
O.030.06
0.42
15.40
0.705.54O.50
O.06
0.104.23
1.28O.060.050.06O.03
O.381.03
0.04
O.72O.161.050.28
0.07
0.040.410.94O.10
0.47
2.47
O.200.07O.19O.32
1.99
16.76
3.95O.26O.39O.770.41O.233.843.28
0.22
3.41
1.32
3.450.140.∞
0。04
5.5l
1.040.39
0.34
0.180.54
1.08O.62
O.45
1.00
4.71
O.5l
1.57O.56
1.00
1.49
O.23
tley5-homtrdey.mha7x3eh664.53.92123
让¥舛¥铂卯鼹毋∞缸配酩矾醪酯酊Cyclohe瑚e,l。ethe“yl・l。methyl.2,4.biB
醯Bicvolo[3,。netehpytl《et—enheney,Dz,-6.寸n删hyl.6-(4一methyl-3.pentenyl)
谚∞n彤瑞心2三3:印咖。’2哪科茹
hylerie-2.(-
4.‘
methy|-3-pentenyl)他钐"巧埘芝黑‘兕嚣警一冀烹1眦吣,龆啊黯灿’25.1ene,[1BR-(laa,4币,7sa,7够,7ha)]
%竹.
methylene..(11-methyletllyl).。(
10l
,
4aa
,
8aa)
鸺锣町缸昭№嚣..,蓼鬣:.-・(-11methylethyl)la・,(,4aa,8阳)-・
∞钳酬0:=:搿。裂4。(5-)..-
l
metllylene-4.hexenyl
)-
跖跖盯船的2。哪州讪絮伽’::,4a,5,6,7,.8‘
。
hydroxy-44a-dimethyl
●
●
∞虬窨{鲐
NatProdResDev0.08
46.08
O.16
0.100.122.850.14
21.0120.稻
22:29l…~
I.58
3.02
480…
0.130.53O.63
s铂…
O.1l
1.150.080.122.18
O.160.20
O.681.24O.08
0.∞
O.34
…
0.070.100.15
O.3l0.07
O.090.9439.18
8.90O.22O.13
0.130.0r7
O.0r7
O.∞
2.70
1.19
O.160.07
O.31O.9l
2.171.98
0.15O.390.44
0.32
O.27
O.26
0.67
1.540.48
0.38
0.740.18
0.29
0.88
O.8l
VoL23
0.485.26
O.60
O.52
…
VoL23
ZHANGSheng-tan.da/:CompositionandAntimicnDbial
Activities
ofEssentialOil
of,-九疵皤Amom/469
94Spathulend34.54695
Caryopylleneo五de
34.672%
(・)・Globulol
34.91l
1H-Cyclopropa[a]naphthalene,la,2,3,5,6,7,7a,7b・
97
octahydro一1.1,7。7a-tetramethyl-,
35.145[1aR-(1aa,7a,7aa,7ha)]-
98Viridifloml
35.424996-acetyl-7・hydmxy-2,2-dimethylbenzopyran
35.560100Caryophylh-3,g(13)-dien-Sa-d
35.78S101
But-3-enal,2-methyl-4・(2,6,6-trimethyl-1-cyclohexenyl)・
37.072
耋|Widdtene
37.2跖
mMuurdol37.760饼a-Cadinol
38.228{宝Cyclohexene.6-ethenyl-6・methyl-1・(1・38.484瞒methylethyl)-3・(I-methylethylldene)・Caryophylla-3,8(13)-dien05卢-ol
38.661研Cm-yophyllenol-II38.904螂.y-Himacha]ene39.402埘w-Santald39.699m
a・Bergamotene40.126m
9.Fluorenone
40.902m(+/・)一(1RS,4aRS,BaRS)-decahydro-5,5,
41.4538a-trimethyl-2一methylene-1一naphthylmethanol
mPhenanthrene42.248卅Neophytadiene
43.809;竺2-Pentadee.anone,6,10,14-re'methyl-
43.943;曼Pentadecandcacid44.785m9.Hexadecenoieacid46.523mDibutylphthalate46.599mHexadecanoicacid
47.075啪
FluorantIIerie
49.001m
Methylheptadecylketone
49.713mPhytol49.831mLinoleicacid50.467m9,17・Octadecadienal,
50.573协9。12-Octadeeadienoic
add(z,z)-
50。584蜥9-OetadecenoiCadd50.650阱Octadecanoicacid
51.006m2-methoxy・10-oxo-5。lla—dlmethyl-6。7,
51.574m1.(3.廿u8te,9n-ytlq卜tm)h,g,d.啦dlpmheetnna。n】【yth旬re.n18e叩mpyp
53.044瑚
2-methyl-3,4-<lihydronaphtlmhne
I删ecello
53.134m
%洲"
53.294耋!2-(Fench-2-y1)fenchane
54.392mTetracoume
54.996m
l-Nonadeeene
56.294
O.18O.77
一
0.12
-O.10
0.07
0.08
O.24
O.260.50
O.23O.20
1.421.021.77
1.37
O.15
O.06
2.90
0.14
5.83
0.160.32
0.080.14
7.90
一
一
0.42O.180.89
O.112.03O.27
O.22O.08O.1l1.470.49
O.1l0.37
0.34
3.150.12
O.09
2.34
1.67
O.23O.33
0.29
O.10
2.68
O.52
5.03
3.7l5.63
0.75
O.铂
16.85
470
NatProdResDevV01.23
.NodetectedAbundance1.4e+071.3e+071e+07800000060000004000000200000011me
.|JlI』.
I。I。.。I^J。
川J
l
13me
5.001000150020002500360035004000450050005500
5.0010.0015.0020.0025.0030.0035.0040.0045.0050.00
Fig.1
TICofseedsbyHydrodislil・laflonmethod
Fig.3
TIC
ofhunksbyHydrodistU-Iafion
method
’,
Abundance1.4e+071.3e+071e+078000000
lIl.1
13me
Fig.2
Table2
6000000
1.LI…It一.“.-l
J
.1
40000002000000Time
t^J‘
JJJ_|IJ上}…。J也
Fig.4
uLk
5.0010001500200025003000350040004500500055005.00la00150020002500300035004000450050005500
TIC
of
seeds
by
B-D
methodTIC
ofhunks
by
B-D
method
CompositionsinthedifferentpartsofFructusAmomibydifferentextractionmethods
Resultsofantibacterialactivitiesofessentialoiltracts
ex-
m/haddifferent
antibacterial
two
activitiesagainstofthetested.Theinhibitionfor
fungi
threefungusand
activityWaSaSsessed
bacterias
extracts
ne
antibacterial
byevaluating
zbnesize
ofdifferentandbacteria
inFig.6.
thepresenceofIZ,MIC
and
MBCvalues.ResultsofIZ
showninFig.5,theinhibitoryeffects
shown
(Table3)showed
thattheessentialoilofFructusAmo—
Table3
Thebacterialinhibition
zone
of
different
extracts(孑±j,删。罪=3)
Bligh,-Dky…erl,“positivc
method(husks)11.0sO.4610.4sO.45
Hydrodi雠illationHydrodistiUationBlish-Dyer
(seeds)
Trichophytonrubrum
14.2±O.6013.9±1.24
(husks)
14.8sO.2513.0sO.79
method(seeds)
13.7sO.5712.6sO.88
………c∞hDl
25.3±0.7532.I±1.10
撕矾啦ytonmemagrophyhm
VoL23
ZHANGShengtandd:Comp031tionandAntlmicmlliH]A。Ilv…%ofEwentbdOil
of胁dⅡ^mmI
47
Indi∞te
no
antibscIerlal
aetivhy
_
0
习0.0融饥'慨fi
‘啪ds’‘nu5‰瘩髫5’
画and
badeHa
Table4
圈。圜。
Fig.6
Effectofessentialoilagainst
Mkr㈣m
gypse-
ⅡⅢ(a)andTdchophymn用6n胛(b)
A
U∞k∞ntml;B—ntia]o】1.C”唧uveconsul.Dpositive
contml
显舞蛊。毖絮乒微船“端髫‘
‘hu8k3’
TheinhibitionzoⅡbofdrfferentextracbforfun-
TheM]CandMBCvaluesforfungiandbacteria(table
4),which
weresensitivetotheessentialoilofFructmAm-
Fig5
omiwcmintherangeof0033and0210mg/mL-_.]O
mg/舢10
000,g/mL
OCO叫mL,respectively
The
MIC
and
MBC
ofdifferentextracIsrn=3
‘m∥mL:。¨∥mL
Conclusion
Theresultsshowed山aEtheessentialoilofFructusAm-om‘haddifierentinhibifionfordlfierentfungiandbac—teriaTheinitibltionforfungibacteria,and
wRs
was
Discussion
In
recent
years,severalresearchels
hadreportedthat
theoxygenatedmono-orsesquiterpenesandmoDo-or
sesquiterpene
betterthanthatfor
bacteria
an—
hydrocarbons
are
the
major
J
components
the
inhibitionforgram—positive
“theessenlialoiIextractedfrom¥everalplantswhichexhibitpotentialantibacterialactivities【7Inliteratures,itshowedhadmonoterpenessuch
theessential
as
betterthanthatforgram・negativebacterianle
fibacterialactivityofessentialoilfromseedswasbetter山an山atfromhusksAndtheantibactetialactivityob—minedthatby
was
oilofthisplant
a—pinene,口-pinene,caryo—
byhydrodistillation
methodwKs
stronger山an
Bll曲一Dyermethod,butthe
Bli曲一Dycr
yieldofessentialoilmethod
lowerthanthatby
㈣tiba【eri日1㈣d
ne.and
borneol
phyHene,cadinene.bergamotene,mymene,d-cuhebe—
as
the
anti
major
components
withstrong
i
icrnhiaI
activitiess,9]Ttlcsc
472
NatProdResD印
VoL23
chemicalcomponents
on
exerttheir
antimicrobial
activity
microorganismsthroughthedisruptionofbacteriaessential
oilsexerts
a
membrane
integrity.…Some
reportedthat
1
11lePharmacopoeiaCommissionthePeople’sRepublicofPress,20Q5,177・177.
of
PRC.Pharmacopoeiaof
Industry
greaterinhibitoryactivityagainstGram—positivebacteri-a.Asitis
Gram-negative
bacteria
are
China.Beijing:Chemical
moreresistanttothethese
plant.based
essentialoil[1l】.and
our
2
HuYL,ZhangZY,Lin
JM.11leresearchofthechemical
on
reports
are
inagreementwithfindings.Inad—
compositionandpharmacological
Fructus
Amoral.JChin
dition,itwasalsopossible
thattheminor
MedMat,2005,28(1),72-74.
3
Mei
components
QX,BiHX.Modem
Pharmacology
Manual.Beijing:Chi・
such嬲limonene,linalooloxide,verbenol,ot・terpineol,myrtenol,famesol,eugenol,caryphyUeneoxide,etc,
mightbeinvolvedinsometypeof
4
na.TraditionalChineseMedicinePress,1999:207.
ThePharmacopoeiaCommissionofPRC.PharmacopoeiaofthePeople’s
antibacterial
syner-
RepublicofChina.Beijing:ChemicalIndustry
sismwithotheractivecomponentsofessentialoil.[12】
Press,2005,AppendixX56.
While。asthedifferencesofkindsand
contentofcorn—
5
MurrayPR,Baron
F_j,PfallerMA,eta1.Manualofclinical
portentsinessentialoilsamples.there锄=esomediffer-
ences
microbiology(6thed.).Washington:ASM.1995.
6
National
Committee
fortheessentialoilextractsininhibitoryactivity.
acetate
f打ClirilealLaboratoryStandard(NC・
susceptibility
tests
Bomyl
and
camphor
are
main
components
in
CI.S).Methods
forbacteriathatA5,New7
fordilutionantimierobial
theessential
oil,andtheirtotalproportioninseedsis
growaerobically.ApprovedStandard,M7-
upto70%,whetherbomylacetateand
camphorplay
Jersey,USA.2000.
a
CakirA,KordaliS,ZenginH,eta/.Compositionandantifun—gal
roleintheantimicrobialprocessremainstobefurtherstudied.Theresultsoftable2andactivitytestshowedthattheantibacterialactivityofessentialoilhadcloselyrelationtothekindsvinyl
activityofessentialoilsisolatedfromHypericumhyssopi。
Hypericum
foti吼and
62-68.
heterophfllum.JFla
Frag,2004,19,
and
relativecontentofthetotalof
containsalkenes,alcohols
8
DebaF,XnanTD,YasudaM,eta1.Chemicalcomposition
and
compositions,which
antioxidant,antibacterialandantifungalactivitiesofthe
Bidenspilosa
l_inn。Vat.Rad/ata.Food
andaldehydes.The
bacterialactivity.
morekinds
and
contentofthees-
essentialoilsfrom
sentialoilcontained,thestrongeritperformedfor
anti—
9
Control,2008,19,346-352.
Qiu
B,Lv
Q,BacFK,税a/.GC-MS
analysis
andantimierobi・
roots
Bligh.Dyer
minoacids
crowave
method‘1
31
isusedtoextracttheLipids.A.
alactivityofessentialoilsfromthefreshanddried
of
and
other
components,and
higher
ultrasonic
or
mi—
10
Codonopsiseordifolioidea.NatProd
449,465.
ResDev,2010,22:445-
methodcouldmake
salne
extractionefficien—
KnobloehK,PauliP,IbetlB,eta1.Antibacterialandantifun—
galproperties
of
cy.Atthe
are
time,thecomponent
damage
and
loss
essentialoilcomponents.JEssentOilRes,
not
easy,while
a
astheorganicsolventextraction,1989,1。603和8.
11
ReynoldsRoyal1996.12
MarinoM,Bersanistudythe
C,Comi
theremayhave
certainextract
ual.Soand
intheactualprocessof
organicsolventsresid・productionandapplica・
the
specificre—
JEF.Martindalethe
extra
Hannacopocia,31sted.
Great
Britain,London.
PharmaceuticalSociety
of
tion,wecouldselectappropriateessentialoilextracts
extraction
methods
accordingto
G.Impedancemeasurements
to
antimierobialactivityofessentialoilsfrom
Lami’
quirements.
而e
acea℃e
contentof出e
essentialoilisdeterminedtobe4%.
195.
essen-
andCompositae.IntJFoodMicrobi01.2001,67,187-
7%inFructusAmomi.whichindicatedthatthetialoilis
all
13耽gll
ED,Dyer
WJ.Arapidmethodfortotalllipidextraction
abundant
naturalresourceforthepharma-
andpurification.CanJBioehemphysiot,1959,37,911-917.
ceuticalindustryandfoodproductionprocessingindus-
田.
天然产物研究与开发NatProdResDel,2011.23:464-472
文章编号:1001-688012011)03-0464-09
中药砂仁挥发油化学成分及其抗茵活性
张生潭1,王兆玉2,汪铁山1,李苗霞1,林敬明h
1南方医科大学珠江医院药剂科,广州510282;2广东药学院生命科学与生物制药学院,广州510006摘要:本研究将Bligh-Dyer溶剂提取法首次应用于植物挥发油的提取,通过GC-MS方法分析,从干燥成熟的砂仁种子和果壳四种挥发油提取物中共鉴定出138种化学成分。挥发油成分主要包括乙酸龙脑酯(5%一47%)、樟脑(4%-17%)、龙脑(1.5%一6%)、莰烯(O.2%一3%)、仅-蒎烯(O.2%一3%)、B・蒎烯(0.2%一5%)以及Ot.柯巴烯(O.1%一2%)等,其中总烯类物质相对含量占总挥发油总成分的10%一40%。从该植物中首次鉴定出蓝桉醇、二环大根香叶烯、薄荷烯醇、二十三烷、二十四烷等新的化学成分35个。所得挥发油对部分真菌及细菌的抑制试验表明其对红色毛癣菌、须毛癣菌、石膏样小孢子癣菌、金黄色葡萄球菌和粪肠球菌均表现出显著的抑制活性。因此,砂仁不仅可作为抗菌药物应用于临床,也可以作为食品原料或天然的食品防腐剂用于食品工业的生产。
关键词:砂仁;挥发油;CC—MS;抗菌活性;BliSh—Dyer法中图分类号:1t284.2
文献标识码:A
CompositionandAntimicrobialActivitiesofEssentialOilofFructusAmomi
ZHANGSheng.talll,WANGZhao.yu2,WANGTie—shahl,LIMiao—xial,LINJing.min91’
1Department
ofPharmacy,ZhujiangHospital,SouthernMedicalUniversity,GuangzhouSl0282,China;
2College矿舻ScienceandBiopharmacology,GuangdongPharmaceutical
Univenity,仇纰删JDD嘶,China
Abstract:’Ik
essentialoil
extractswere
respectivdyobtainedfromthes∞d8andhusksofAmomumvillosuml_ourby
u-
sing
Bligh・Dyermethodsfor
thefirsttime.Onehundredandtllirty-eightcompositions
were
identifiedbyGC・MS
from
fouressentialoilextracts.,11lemaincomponentsoftheessentialoilwerebomylacetate,camphor,bomeol,俚-pincne,13・
pinene
and
d.eopalene.etc.The
relative
contentsoftotalalkenes
and
ketonescomposition
accountedfor10%40%in
integralofessentialoil.Thirty・fivecompositionssuch
a8
neophytadiene,globulol,bicyclogermacrene,etc
were
identified
fromthis
plant铀dy.‰resultsofinhibitory
effect
forbacteriaandfun#showedthattheessentialodhadremarkable
inhibitoryeffectsforTrichophytonrubnlm,Tfichophytonmentagrophyton,Micresporumgypseum,Staphyleccecus
811reus,
andEntemcoccusfaecalls.Ther商ore.the
essential
oilCanbeusedinthefoodindustry
a8rswfoodmaterial
or
natural
foodpreservation.
Keywords:FructusAmom/;essentialoil;C,C--MS;antibacterialactivity;Bligh-Dyermethod
spleenand
antidiarrhea,etc.[21Inaddition,itisalso
a
keyflavoringinbothfood
anddrink.Theseactivities
Amom/istheripenedf,抛itsofseveralphntsofhavebeenlinked
to
thepresenceofessentialoil.L3J
Amomum(Zingiberaceae)includingA.villosum
Inthisstudy,allessentialoilwasextractedfrom
the
Lour.Vat.xanthioidesT.L.Wu
et
seedsandhusksofFructusAmom/byusingBligh・DyerandA.IongiligulareT.L.Wu.…Ithasbeen
methodandHydrodistillationmethodrespectively.Oneas
an
ingedientintraditionalchinesemedicine
hundredandthirty・eightcompositionswereanalyzedeliminatingdampness,promotingappetite,warming
andidentifiedbyGC—MSfromthefoursamples.ni啊・
fivecompositionssuch
as
neophytadiene,globulol,ger-
macrene
Received
August
26.2010;AcceptedDecember31。2010
D,fluorenone,bicyclogermacrene,piperitol,
FoundationItem:This
pmjcctW85鲫pponedbyGuant弘houMunicipal
tricosane。tetracosane。etc
were
firstidentified
inthis
ScienceandTechnologySupportProgramme(2008Zl—FA61)
toconventional・Corresponding.author
Tel:86-20-61643557;E—mllil:linjml231@210n.c锄
plant.Comparedhydrodistillationmeth-
Introduction
Fructusgenus
Lour,A.villosum
Senjen
used
for
VoL23
ZHANGSheng—tan。eta/:CompositionandAnfimicrobialActivitiesofEssentialOnofFructusAmom/465
od.21kindsofnewchemicalcompositionswereidenti.
fled.AndtheextractionefficiencyofBligh—Dyermethod
ismuch
higherthanthatofhydrodistillation
method.
ne
resultsofpharmacologicalactivityshowedthatthe
essentialoilofFructusAmomihad
significant
inhibitory
effectsforchosenbacteriaandfungi.Inthispaper,a
new
applicationforFructusAmomiisprovidedinanti—
biotics
and
foodpreservation,whilerelated
scientifictheoryandexperimentalevidenceare
alsopresentedforutilizingessentialoilofFructusAmomiinthephanna.ceuticalindustryandfoodindustryofdevelopmentandutilizatio玎.
Materials
and
Methods
General
GC-MSsystem(GC6890seriesII;MSD5973,Hew.
1ett
Packard,America),RotaryEvaporator(R-200,
BUCHI
company,Switzerland),ultrasoniccleaning
e・
quipment(KQ-600DE,KunshanUltrasonicequipment
company,China),electronicscales(BL-2000S,setracompany,America),Incubator(DNP-9162,Ningbo
SoutheastInstrumentCompany,China),biologicalsafe—
ty
cabinet(BHC.1300IIA/B3,Suzhou
Antai
Air’rech
Co,Ltd,China);Antimicrobialsusceptibility
test
discs(5×50discs,OxoidLtd,England),Medium(OxoidLtd,England).PlantMaterials
Ourresearchwascarried
out
on
FructusAmom/,inclu—
dingseeds
and
husksofdryingandripeningfruits.The
FructusAmomiwaspurchased
in
Guangzhou
herbal
company.Theoriginalpurchasingplacewas
Guang
dong
province,China(.Batch
number:
Materials
rrichophytonrubrumCMCC(F)TIa,Trichophyton
mell—
CMCC(F)T5a,andMicrosporum
gypseum
were
purchasedinSkinResearchInsti.
ofChineseAcademyofMedicalSciences.Esche・
coliATCC35218,StaphyloccocusaureR¥Rosen-ATCC29213,Enterococcusfaeeal括ATCC29212
Enterobacter
cloacaeATCC700323wer@providedMedicalTestingCenter,Zhujiang
hospital.
andanalysisoftheessentialoil
HydrodistillationmethodSeeds
andhusks
ofFructus
Amomiwerepeeledand
crushedseparately,with24meshscreen.Onehundredgramsof
plant
materialsweredistilledwith
water(1
L)fh
6hin
an
extractionequipmentinaccordance
withthedescriptionofthePharmacopoeiaofthePeo-pie’sRepublicof
China(2005)E4J.弧e
essentialoil
was
collectedanddried
over
anhydrousNa2S04,then
storedinsealedvialsat4℃.
居2趣出一助r删肠d
卟e
plant
materials(100g)and500mLofextracting
solvent(the
volumeratioofmethanol:chloroform:wa.
ter
was1:l:0.5)wereplacedina
Erlenmayerflask
(1500mL)andsoakedforthreedays.Andthenthe
flaskwastreatedthreetimeswith
ultrasonic(40kHz)
at
40±l℃f.or45min..11le
extracts
werecentrifuged
and
thesupernatantswerecombined.After
evaporation
ofthesolvent,theessentialoil
wascollectedandstored
砒4℃.
Gaschromatography-massspectrometryanalysis
弧e
essentialoilextractswereanalyzedbygas・chroma-
tographieally.’IheC,Coperatingconditionswere
as
fol-
lows:columntemperaturewasincreasedfrom50to250
℃at
arateof
5'U/min;injectortemperaturewas
set
to
250℃.andthedetector
temperaturewassetto280
℃:thecarriergas
was
He
at
a
rateof1.0mL/min.
’11leMSoperatingconditionswere船follows:carriergas
was
Heat
arate
of1.0mL/min;massspectrometryi-
onization
energy
was70
eV,and
electron
multiplier
tubevoltagewas
1905V;theionsourcetemperature
was
setto
230℃.andthequadrupoletemperature
was
setto
150℃ithemass
scan
range
was30-550amu.
’nleethylacetatesolutionoftheessentialoilwasdis-
solvedwith
1曲EtOAcandinjectedinsplitmode.
711lecompositionsinessentialoilwere
identifiedbyGC-MSbymassfragmentationpattern
and
spectralcompar-isonwithstandardsintheWiley275and
Nist05librar-
ies.
Determinationofantibacterialactivity
Invitroantibacterialactivitiesoftheessentialoil
ex.
wereevaluatedbydiscdiffusionmethodusing
Mueller-HintonAgarforbacteriaandSabouraud
Agar
for
fungi
with
determinationof
inhibition
zones
medicine20090601).
Microbiologicaltagrophyton
CMCC(F)M2a
tute
richiatracts
bachandbyPreparation
NatProdResD朗
VoL23
(Iz).‘51A100trLsuspensionoftestedbacteriaand
onon
organismsaftermacroscopicevaluation,wasdetermined
fungi(108CFU/mL)spreadedAgarmediums.106
innl
嬲MIC.whichwasexpressedinms/mL.DeterminationofMinimumBactericidaltration(M吣C)
TheMBCofessentialoilweredeterminedaccordingtotheMICvalues.Ofeachwellshowingcompleteabsenceof
Concen.
止of
and
essentialoilwasdropped
on
filterpapem
place
theAgarsurface.Standard
as
antibioticand
blackcontroldiskswereused
reference.Plateswere
incubatedat28℃for96hforfungiandat37℃for24hforbacteria,respectively.Theinhibiton
zones
were
growth,20灿was
transferred
to
agarplatesandin—24hat37℃for
es-
measuredi/i
diameters.The
nLrn
scaleofmeasurementwas
was
cubated72hat28oCforfungi
and
thefollowing:>15lO-15mm
zone
zone
stronglyinhibitory,
toni
bacteria.respectively.’11lelowestconcentrationofsentialoilwhich
no
moderately
inhibitory.and<10
viablebacteria
or
fungi
wereidenri—
,
zoneweaklyinhibitory.
fledwastheMBC.
of船nhn啪In蜥呻Co眦嘶昀。De协粤曲ption(MIC)
The
MICweremeasuredbymicrodilurionRPMI一1640
assay
recommended
by
NCCLS
susceptibility
ResuIts锄d……一一………conclusio璐
Chemicalcompositionsoftheessentialoilextracts
Onehundredandthiay・・eightcompositionswereidenri・-fledfromthefouressentialoilsamples(Table1).Theyieldsofessentialoilfromseedsdistillationmethodwere
(2000).161%e
wellconmined
96wellmicroplatewasused.each
bacteria
RPMI-1640
liquid
10止of
test
and
husksbyhydro.
medium(bacteriamL)and
concentrationof0.5-5×105CFU/
essentialoilsolution.10斗L
3.57%(g/g)and0.97%
thosebyBligh—Oyermethod
90止of
(s/g),respectively,while
were
sterilenormalsalinewasblackcontr01.Anappropriate
7.01%(s/g)and5.37%(s/g),respectively.
maincompositionsofessentialoilwerebomyl
ace.
amount
ofTween-80wasaddedtomakeitinto
a
5%
The
solution。ThisconcentrationofTween.80didnotaffectthegrowthofbacteria
rate,camphor,borneol,ot—pinene,伊pinene,and
other
and
fungi.11】efinalconcentra—components.%e
and
69.85%in
relativecontentsofcompositionsiden-
fionsoftheessentialoilinwellsrangedfrom0.00977rifledwereaccountedfor93.39%。92.18%。74.85%,
fouressentialoils,respectively,and
compositionswere
TIC
ofoil
ms/mL
were
to
72h
ms/mL.’rheincubationconditions
at28℃forfungiand24hat37℃forbac-
10.000
thoseoftotalalkenesandketonesaccountedfor10%-40%(TablesamplesshowninFigure1—4.
teria,respectively,andeachtreatmentwasduplicate
performed
in
2).The
thrice.ne
lowestconcentrationofthetest
samples,whichdidnotshow
Table1
anyvisualgrowthoftested
ChemicalconstituentsoftheessentialoilextractsofFructusAmomi
Ethylbenzenep-Xylcne
4.9635.1596.3206.3686.4536.5817.0197.1047.S78
0.020.221.48O.02
一
O.19O.66
一
Tricydo[2
21
0(2,6)]heptane,1,7,7-trimethyl-
O.cr7
●
Bicycio[310】hexane,4-methyl・1・(1-methylethyl)・
a-Thujene
a・Pina坼CampheneVerbeneneSabinene
0.7l
0.402.26O.270.090.24
2.0r70.2lO.2l
一
0.462.43
●
一
0.∞
、roL23
ZHANGSheng-tan。da/:CompositionandAnfimierobialActivitiesofEssentialOilofFru斌uaAmom/467
10口-Pine舱
7.73011Bicyclo[3
1
1】heptane,6,6-dimethyl-2-methylene-
7.73212口・Myrcene7.985131.Phellandrene8.49614a-Terpinene
8.78615Benzene,l-methyl-2・(1-methylethyl)
9.024161-1sopropyl-4-methylbenzene
9.03017Limonend
9.12518卢-Thujene
9.200191-Terplnene9.95320Linalooloxide
10.32921ds爷-Terpineol
10.37622n薯n8-Sahinenehydrate
10.38123Terpinolene
10.71924口-Campholenealdehyde
10.85525Benzene。1-methyl-4-(1・methylethenyl)・
10.91426Linalool
11.Z驺
27ci8・Sabinenehydrate
11.”7
281.6-OetMien-3-oi,3。7--climethyl-
11.28529Bicyclo[3
1
1]heptan-2-one,6,6-dimethyl
11.325301,5,7-Octauien-3-oi,3,7-dimethyl-
11.34731a-Thujone
11.70232D-Fenchylalcohol
11.88033(4-)-Nopinone
12.34334Bicyelo[3
1
1]heptan-3..ol,6,6-dimethyl-2-methylene-
12.45735hans.Pinocarveol
12.55636Camphor12.67037a.Phellandren-8-01.
12.74638a-Thujone12.89439
2(i0)-Pinen-3-one
12.979柏lsoborneol
13.12041Camphol
13.405423-Cyclohexen-I-ol。4-methyl-1-(1一methylethyl)-
13.57143
4-Terpinenol
13.641
44Bieyclo[3ll】hept-2-ene-2-carboxa
ldehyde,6,6-dimethyl・
13.92845Myrtenal
13.96746Bieyclo[3
l
l】hept-2-ene-2一methanol,6,6-dimethyl・
14.02847Myrtenol
14.10448Bicyclo[3ll】hept-3-en-2-one,4,6,6-trimethyl・。(1s)・
14.28349Verben帆e
14.3295011'1m8・Piperltol
14.41251
n哪hyl
acetate
14.490
O.2l
0.630.06O.10
O.030.06
0.42
15.40
0.705.54O.50
O.06
0.104.23
1.28O.060.050.06O.03
O.381.03
0.04
O.72O.161.050.28
0.07
0.040.410.94O.10
0.47
2.47
O.200.07O.19O.32
1.99
16.76
3.95O.26O.39O.770.41O.233.843.28
0.22
3.41
1.32
3.450.140.∞
0。04
5.5l
1.040.39
0.34
0.180.54
1.08O.62
O.45
1.00
4.71
O.5l
1.57O.56
1.00
1.49
O.23
tley5-homtrdey.mha7x3eh664.53.92123
让¥舛¥铂卯鼹毋∞缸配酩矾醪酯酊Cyclohe瑚e,l。ethe“yl・l。methyl.2,4.biB
醯Bicvolo[3,。netehpytl《et—enheney,Dz,-6.寸n删hyl.6-(4一methyl-3.pentenyl)
谚∞n彤瑞心2三3:印咖。’2哪科茹
hylerie-2.(-
4.‘
methy|-3-pentenyl)他钐"巧埘芝黑‘兕嚣警一冀烹1眦吣,龆啊黯灿’25.1ene,[1BR-(laa,4币,7sa,7够,7ha)]
%竹.
methylene..(11-methyletllyl).。(
10l
,
4aa
,
8aa)
鸺锣町缸昭№嚣..,蓼鬣:.-・(-11methylethyl)la・,(,4aa,8阳)-・
∞钳酬0:=:搿。裂4。(5-)..-
l
metllylene-4.hexenyl
)-
跖跖盯船的2。哪州讪絮伽’::,4a,5,6,7,.8‘
。
hydroxy-44a-dimethyl
●
●
∞虬窨{鲐
NatProdResDev0.08
46.08
O.16
0.100.122.850.14
21.0120.稻
22:29l…~
I.58
3.02
480…
0.130.53O.63
s铂…
O.1l
1.150.080.122.18
O.160.20
O.681.24O.08
0.∞
O.34
…
0.070.100.15
O.3l0.07
O.090.9439.18
8.90O.22O.13
0.130.0r7
O.0r7
O.∞
2.70
1.19
O.160.07
O.31O.9l
2.171.98
0.15O.390.44
0.32
O.27
O.26
0.67
1.540.48
0.38
0.740.18
0.29
0.88
O.8l
VoL23
0.485.26
O.60
O.52
…
VoL23
ZHANGSheng-tan.da/:CompositionandAntimicnDbial
Activities
ofEssentialOil
of,-九疵皤Amom/469
94Spathulend34.54695
Caryopylleneo五de
34.672%
(・)・Globulol
34.91l
1H-Cyclopropa[a]naphthalene,la,2,3,5,6,7,7a,7b・
97
octahydro一1.1,7。7a-tetramethyl-,
35.145[1aR-(1aa,7a,7aa,7ha)]-
98Viridifloml
35.424996-acetyl-7・hydmxy-2,2-dimethylbenzopyran
35.560100Caryophylh-3,g(13)-dien-Sa-d
35.78S101
But-3-enal,2-methyl-4・(2,6,6-trimethyl-1-cyclohexenyl)・
37.072
耋|Widdtene
37.2跖
mMuurdol37.760饼a-Cadinol
38.228{宝Cyclohexene.6-ethenyl-6・methyl-1・(1・38.484瞒methylethyl)-3・(I-methylethylldene)・Caryophylla-3,8(13)-dien05卢-ol
38.661研Cm-yophyllenol-II38.904螂.y-Himacha]ene39.402埘w-Santald39.699m
a・Bergamotene40.126m
9.Fluorenone
40.902m(+/・)一(1RS,4aRS,BaRS)-decahydro-5,5,
41.4538a-trimethyl-2一methylene-1一naphthylmethanol
mPhenanthrene42.248卅Neophytadiene
43.809;竺2-Pentadee.anone,6,10,14-re'methyl-
43.943;曼Pentadecandcacid44.785m9.Hexadecenoieacid46.523mDibutylphthalate46.599mHexadecanoicacid
47.075啪
FluorantIIerie
49.001m
Methylheptadecylketone
49.713mPhytol49.831mLinoleicacid50.467m9,17・Octadecadienal,
50.573协9。12-Octadeeadienoic
add(z,z)-
50。584蜥9-OetadecenoiCadd50.650阱Octadecanoicacid
51.006m2-methoxy・10-oxo-5。lla—dlmethyl-6。7,
51.574m1.(3.廿u8te,9n-ytlq卜tm)h,g,d.啦dlpmheetnna。n】【yth旬re.n18e叩mpyp
53.044瑚
2-methyl-3,4-<lihydronaphtlmhne
I删ecello
53.134m
%洲"
53.294耋!2-(Fench-2-y1)fenchane
54.392mTetracoume
54.996m
l-Nonadeeene
56.294
O.18O.77
一
0.12
-O.10
0.07
0.08
O.24
O.260.50
O.23O.20
1.421.021.77
1.37
O.15
O.06
2.90
0.14
5.83
0.160.32
0.080.14
7.90
一
一
0.42O.180.89
O.112.03O.27
O.22O.08O.1l1.470.49
O.1l0.37
0.34
3.150.12
O.09
2.34
1.67
O.23O.33
0.29
O.10
2.68
O.52
5.03
3.7l5.63
0.75
O.铂
16.85
470
NatProdResDevV01.23
.NodetectedAbundance1.4e+071.3e+071e+07800000060000004000000200000011me
.|JlI』.
I。I。.。I^J。
川J
l
13me
5.001000150020002500360035004000450050005500
5.0010.0015.0020.0025.0030.0035.0040.0045.0050.00
Fig.1
TICofseedsbyHydrodislil・laflonmethod
Fig.3
TIC
ofhunksbyHydrodistU-Iafion
method
’,
Abundance1.4e+071.3e+071e+078000000
lIl.1
13me
Fig.2
Table2
6000000
1.LI…It一.“.-l
J
.1
40000002000000Time
t^J‘
JJJ_|IJ上}…。J也
Fig.4
uLk
5.0010001500200025003000350040004500500055005.00la00150020002500300035004000450050005500
TIC
of
seeds
by
B-D
methodTIC
ofhunks
by
B-D
method
CompositionsinthedifferentpartsofFructusAmomibydifferentextractionmethods
Resultsofantibacterialactivitiesofessentialoiltracts
ex-
m/haddifferent
antibacterial
two
activitiesagainstofthetested.Theinhibitionfor
fungi
threefungusand
activityWaSaSsessed
bacterias
extracts
ne
antibacterial
byevaluating
zbnesize
ofdifferentandbacteria
inFig.6.
thepresenceofIZ,MIC
and
MBCvalues.ResultsofIZ
showninFig.5,theinhibitoryeffects
shown
(Table3)showed
thattheessentialoilofFructusAmo—
Table3
Thebacterialinhibition
zone
of
different
extracts(孑±j,删。罪=3)
Bligh,-Dky…erl,“positivc
method(husks)11.0sO.4610.4sO.45
Hydrodi雠illationHydrodistiUationBlish-Dyer
(seeds)
Trichophytonrubrum
14.2±O.6013.9±1.24
(husks)
14.8sO.2513.0sO.79
method(seeds)
13.7sO.5712.6sO.88
………c∞hDl
25.3±0.7532.I±1.10
撕矾啦ytonmemagrophyhm
VoL23
ZHANGShengtandd:Comp031tionandAntlmicmlliH]A。Ilv…%ofEwentbdOil
of胁dⅡ^mmI
47
Indi∞te
no
antibscIerlal
aetivhy
_
0
习0.0融饥'慨fi
‘啪ds’‘nu5‰瘩髫5’
画and
badeHa
Table4
圈。圜。
Fig.6
Effectofessentialoilagainst
Mkr㈣m
gypse-
ⅡⅢ(a)andTdchophymn用6n胛(b)
A
U∞k∞ntml;B—ntia]o】1.C”唧uveconsul.Dpositive
contml
显舞蛊。毖絮乒微船“端髫‘
‘hu8k3’
TheinhibitionzoⅡbofdrfferentextracbforfun-
TheM]CandMBCvaluesforfungiandbacteria(table
4),which
weresensitivetotheessentialoilofFructmAm-
Fig5
omiwcmintherangeof0033and0210mg/mL-_.]O
mg/舢10
000,g/mL
OCO叫mL,respectively
The
MIC
and
MBC
ofdifferentextracIsrn=3
‘m∥mL:。¨∥mL
Conclusion
Theresultsshowed山aEtheessentialoilofFructusAm-om‘haddifierentinhibifionfordlfierentfungiandbac—teriaTheinitibltionforfungibacteria,and
wRs
was
Discussion
In
recent
years,severalresearchels
hadreportedthat
theoxygenatedmono-orsesquiterpenesandmoDo-or
sesquiterpene
betterthanthatfor
bacteria
an—
hydrocarbons
are
the
major
J
components
the
inhibitionforgram—positive
“theessenlialoiIextractedfrom¥everalplantswhichexhibitpotentialantibacterialactivities【7Inliteratures,itshowedhadmonoterpenessuch
theessential
as
betterthanthatforgram・negativebacterianle
fibacterialactivityofessentialoilfromseedswasbetter山an山atfromhusksAndtheantibactetialactivityob—minedthatby
was
oilofthisplant
a—pinene,口-pinene,caryo—
byhydrodistillation
methodwKs
stronger山an
Bll曲一Dyermethod,butthe
Bli曲一Dycr
yieldofessentialoilmethod
lowerthanthatby
㈣tiba【eri日1㈣d
ne.and
borneol
phyHene,cadinene.bergamotene,mymene,d-cuhebe—
as
the
anti
major
components
withstrong
i
icrnhiaI
activitiess,9]Ttlcsc
472
NatProdResD印
VoL23
chemicalcomponents
on
exerttheir
antimicrobial
activity
microorganismsthroughthedisruptionofbacteriaessential
oilsexerts
a
membrane
integrity.…Some
reportedthat
1
11lePharmacopoeiaCommissionthePeople’sRepublicofPress,20Q5,177・177.
of
PRC.Pharmacopoeiaof
Industry
greaterinhibitoryactivityagainstGram—positivebacteri-a.Asitis
Gram-negative
bacteria
are
China.Beijing:Chemical
moreresistanttothethese
plant.based
essentialoil[1l】.and
our
2
HuYL,ZhangZY,Lin
JM.11leresearchofthechemical
on
reports
are
inagreementwithfindings.Inad—
compositionandpharmacological
Fructus
Amoral.JChin
dition,itwasalsopossible
thattheminor
MedMat,2005,28(1),72-74.
3
Mei
components
QX,BiHX.Modem
Pharmacology
Manual.Beijing:Chi・
such嬲limonene,linalooloxide,verbenol,ot・terpineol,myrtenol,famesol,eugenol,caryphyUeneoxide,etc,
mightbeinvolvedinsometypeof
4
na.TraditionalChineseMedicinePress,1999:207.
ThePharmacopoeiaCommissionofPRC.PharmacopoeiaofthePeople’s
antibacterial
syner-
RepublicofChina.Beijing:ChemicalIndustry
sismwithotheractivecomponentsofessentialoil.[12】
Press,2005,AppendixX56.
While。asthedifferencesofkindsand
contentofcorn—
5
MurrayPR,Baron
F_j,PfallerMA,eta1.Manualofclinical
portentsinessentialoilsamples.there锄=esomediffer-
ences
microbiology(6thed.).Washington:ASM.1995.
6
National
Committee
fortheessentialoilextractsininhibitoryactivity.
acetate
f打ClirilealLaboratoryStandard(NC・
susceptibility
tests
Bomyl
and
camphor
are
main
components
in
CI.S).Methods
forbacteriathatA5,New7
fordilutionantimierobial
theessential
oil,andtheirtotalproportioninseedsis
growaerobically.ApprovedStandard,M7-
upto70%,whetherbomylacetateand
camphorplay
Jersey,USA.2000.
a
CakirA,KordaliS,ZenginH,eta/.Compositionandantifun—gal
roleintheantimicrobialprocessremainstobefurtherstudied.Theresultsoftable2andactivitytestshowedthattheantibacterialactivityofessentialoilhadcloselyrelationtothekindsvinyl
activityofessentialoilsisolatedfromHypericumhyssopi。
Hypericum
foti吼and
62-68.
heterophfllum.JFla
Frag,2004,19,
and
relativecontentofthetotalof
containsalkenes,alcohols
8
DebaF,XnanTD,YasudaM,eta1.Chemicalcomposition
and
compositions,which
antioxidant,antibacterialandantifungalactivitiesofthe
Bidenspilosa
l_inn。Vat.Rad/ata.Food
andaldehydes.The
bacterialactivity.
morekinds
and
contentofthees-
essentialoilsfrom
sentialoilcontained,thestrongeritperformedfor
anti—
9
Control,2008,19,346-352.
Qiu
B,Lv
Q,BacFK,税a/.GC-MS
analysis
andantimierobi・
roots
Bligh.Dyer
minoacids
crowave
method‘1
31
isusedtoextracttheLipids.A.
alactivityofessentialoilsfromthefreshanddried
of
and
other
components,and
higher
ultrasonic
or
mi—
10
Codonopsiseordifolioidea.NatProd
449,465.
ResDev,2010,22:445-
methodcouldmake
salne
extractionefficien—
KnobloehK,PauliP,IbetlB,eta1.Antibacterialandantifun—
galproperties
of
cy.Atthe
are
time,thecomponent
damage
and
loss
essentialoilcomponents.JEssentOilRes,
not
easy,while
a
astheorganicsolventextraction,1989,1。603和8.
11
ReynoldsRoyal1996.12
MarinoM,Bersanistudythe
C,Comi
theremayhave
certainextract
ual.Soand
intheactualprocessof
organicsolventsresid・productionandapplica・
the
specificre—
JEF.Martindalethe
extra
Hannacopocia,31sted.
Great
Britain,London.
PharmaceuticalSociety
of
tion,wecouldselectappropriateessentialoilextracts
extraction
methods
accordingto
G.Impedancemeasurements
to
antimierobialactivityofessentialoilsfrom
Lami’
quirements.
而e
acea℃e
contentof出e
essentialoilisdeterminedtobe4%.
195.
essen-
andCompositae.IntJFoodMicrobi01.2001,67,187-
7%inFructusAmomi.whichindicatedthatthetialoilis
all
13耽gll
ED,Dyer
WJ.Arapidmethodfortotalllipidextraction
abundant
naturalresourceforthepharma-
andpurification.CanJBioehemphysiot,1959,37,911-917.
ceuticalindustryandfoodproductionprocessingindus-
田.