中药砂仁挥发油化学成分及其抗菌活性

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

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

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

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

O.19O.66

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

0.462.43

0.∞

、roL23

ZHANGSheng-tan。da/:CompositionandAnfimierobialActivitiesofEssentialOilofFru斌uaAmom/467

10口-Pine舱

7.73011Bicyclo[3

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]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]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】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-)..-

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

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

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

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

.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

_

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)

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

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

icrnhiaI

activitiess,9]Ttlcsc

472

NatProdResD印

VoL23

chemicalcomponents

on

exerttheir

antimicrobial

activity

microorganismsthroughthedisruptionofbacteriaessential

oilsexerts

membrane

integrity.…Some

reportedthat

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

HuYL,ZhangZY,Lin

JM.11leresearchofthechemical

on

reports

are

inagreementwithfindings.Inad—

compositionandpharmacological

Fructus

Amoral.JChin

dition,itwasalsopossible

thattheminor

MedMat,2005,28(1),72-74.

Mei

components

QX,BiHX.Modem

Pharmacology

Manual.Beijing:Chi・

such嬲limonene,linalooloxide,verbenol,ot・terpineol,myrtenol,famesol,eugenol,caryphyUeneoxide,etc,

mightbeinvolvedinsometypeof

na.TraditionalChineseMedicinePress,1999:207.

ThePharmacopoeiaCommissionofPRC.PharmacopoeiaofthePeople’s

antibacterial

syner-

RepublicofChina.Beijing:ChemicalIndustry

sismwithotheractivecomponentsofessentialoil.[12】

Press,2005,AppendixX56.

While。asthedifferencesofkindsand

contentofcorn—

MurrayPR,Baron

F_j,PfallerMA,eta1.Manualofclinical

portentsinessentialoilsamples.there锄=esomediffer-

ences

microbiology(6thed.).Washington:ASM.1995.

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.

CakirA,KordaliS,ZenginH,eta/.Compositionandantifun—gal

roleintheantimicrobialprocessremainstobefurtherstudied.Theresultsoftable2andactivitytestshowedthattheantibacterialactivityofessentialoilhadcloselyrelationtothekindsvinyl

activityofessentialoilsisolatedfromHypericumhyssopi。

Hypericum

foti吼and

62-68.

heterophfllum.JFla

Frag,2004,19,

and

relativecontentofthetotalof

containsalkenes,alcohols

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—

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

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

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

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

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

O.19O.66

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

0.462.43

0.∞

、roL23

ZHANGSheng-tan。da/:CompositionandAnfimierobialActivitiesofEssentialOilofFru斌uaAmom/467

10口-Pine舱

7.73011Bicyclo[3

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]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]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】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-)..-

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

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

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

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

.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

_

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)

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

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

icrnhiaI

activitiess,9]Ttlcsc

472

NatProdResD印

VoL23

chemicalcomponents

on

exerttheir

antimicrobial

activity

microorganismsthroughthedisruptionofbacteriaessential

oilsexerts

membrane

integrity.…Some

reportedthat

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

HuYL,ZhangZY,Lin

JM.11leresearchofthechemical

on

reports

are

inagreementwithfindings.Inad—

compositionandpharmacological

Fructus

Amoral.JChin

dition,itwasalsopossible

thattheminor

MedMat,2005,28(1),72-74.

Mei

components

QX,BiHX.Modem

Pharmacology

Manual.Beijing:Chi・

such嬲limonene,linalooloxide,verbenol,ot・terpineol,myrtenol,famesol,eugenol,caryphyUeneoxide,etc,

mightbeinvolvedinsometypeof

na.TraditionalChineseMedicinePress,1999:207.

ThePharmacopoeiaCommissionofPRC.PharmacopoeiaofthePeople’s

antibacterial

syner-

RepublicofChina.Beijing:ChemicalIndustry

sismwithotheractivecomponentsofessentialoil.[12】

Press,2005,AppendixX56.

While。asthedifferencesofkindsand

contentofcorn—

MurrayPR,Baron

F_j,PfallerMA,eta1.Manualofclinical

portentsinessentialoilsamples.there锄=esomediffer-

ences

microbiology(6thed.).Washington:ASM.1995.

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.

CakirA,KordaliS,ZenginH,eta/.Compositionandantifun—gal

roleintheantimicrobialprocessremainstobefurtherstudied.Theresultsoftable2andactivitytestshowedthattheantibacterialactivityofessentialoilhadcloselyrelationtothekindsvinyl

activityofessentialoilsisolatedfromHypericumhyssopi。

Hypericum

foti吼and

62-68.

heterophfllum.JFla

Frag,2004,19,

and

relativecontentofthetotalof

containsalkenes,alcohols

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—

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

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.


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