全 文 ：天然产物研究与开发 NatProdResDev2007, 19:822-826
文章编号:1001-6880(2007)05-0822-05
ReceivedApril20, 2006;AcceptedJune30, 2006
＊CorrespondingauthorTel:86-511-8780226;E-mail:zhangzhicai2001@
yahoo.com.cn
利用 GC-MS技术分析金耳发酵液挥发性成分
张志才 1＊ ,章克昌 2 ,俞建国 3 ,顾立新 3
1江苏大学食品与生物工程学院 ,镇江 212013;
2江南大学生物工程学院生物资源实验室 ,无锡 214036;3江苏泰兴同原堂生物工程公司 , 泰兴 225400
摘 要:利用 GC-MS技术对金耳发酵液中挥发性风味物质进行了分析。样品成分的质谱图与数据库(Nist库
andWiley库)中已知化合物质谱图比较。该研究证明金耳发酵液中匹配指数在 800以上的倍半萜类化合物约
占整个挥发性成分 47%。
关键词:金耳;顶空进样;气相色谱-质谱技术;匹配指数;挥发性香味物质
中图分类号:R284.2 文献标识码:A
AnalysisoftheVolatileCompoundsin
TremelaaurantialbaBrothbyGC-MSMethod
ZHANGZhi-cai1＊ , ZHANGKe-chang2 , YUJian-guo3 , GULi-xin3
1SchoolofFoodScienceandBiotechnology, JiangsuUniversity, Zhenjiang212013 , China;
2LabofBiomassResources, SchoolofBiotechnology, SouthernYangtzeUniversity, Wuxi214036 , China;
3JiangsuTaixingTongyuantangBio-technologicalCorporation, Taixing225400 , China
Abstract:ThevolatileflavorinTremellaaurantialbafermentationbrothwasinvestigatedbyhead-spacegaschromatogra-
phy-massspectrometry(GC-MS).Massspectraldataofthesamplecomponentswerefoundtomatchupwiththoseof
knowncompoundsinadatabase(NistdatabaseandWileydatabase).Thematchindexesof34sesquiterpenesaremore
than800.Theidentifiedsesquiterpenesaccountedformorethan47% ofthetotalflavorcompounds.
Keywords:Tremelaaurantialba;head-space;GC-MS;matchindex;volatileflavour
Introduction
ThegenusTremelaPers.(Tremelales, HigherBasidio-
mycetes)belongstotheso-caled`JelyMushrooms
groupthatform gelatinousfruitingbodies.About80
specieshavebeenidentifiedandnoneofthem are
knowntobepoisonous[ 1] .Jelymushroomsfromthe
genusTremelahavebeenusedasfoodandtraditional
medicinesforcenturiesinmanyAsiancountries.Dur-
ingthepastdecadestheyhavebeenextensivelyinvesti-
gatedfortheirmedicinalefectsinvivoandinvitro.
Clinicalapplicationsweredeveloped fordiferent
groupsofpatients.T.aurantialbafruit-bodypolysac-
charide(TFP)possesestheabilitiesofanti-diabetic
andglucoselowingefect[ 2] .PolysaccharidesfromT.
aurantialbamycelium(TMP)werereportedtoprevent
orreleasethediet-inducedhyperlipidemia[ 3] andal-
loxan-inducedhyperglycermia.T.aurantialba fruit-
bodydemonstratedtheefectsofanti-inflammation[ 4] ,
promotesalivasecretion, moistenlungs[ 5] andstop
cough[ 6] , decreaseitchinginthethroat[ 7] andin-
creaseimmunity[ 5, 8] .ThefermentedproductsfromT.
aurantialbainhibitsignificantlyformationofthrombus,
theperoxidelipidfromdecomposingintoMDAinbrain
tissueandplateletaggregationinducedbyADPinnor-
malrate[ 9] .TheextractfromT.aurnatialbahadbeen
proventodecreasethelevelofplasmglucose, fruc-
tosamineandtriglycerides[ 10] However, theingredients
besidespolysaccharideofT.aurantialbahavenotbeen
investigated.
Sesquiterpenesisalargegroupofcompoundswithvari-
ousandimportantbio-function, suchashepatoprotec-
DOI :10.16333/j.1001-6880.2007.05.025
tive[ 11] , anti-ulcerogenic[ 12] , antifedant[ 13, 14] , autoxi-
dation[ 15] , anti-tumor[ 16] andantimicrobialactivities.
Inthepresentstudy, itwasfoundthatsesquiterpenes
couldbeformedinthesubmerged-culturedbrothofT.
aurantialba, andmorethan34 kindsofsesquiterpenes
inT.aurantialbasubmerged-culturedbrothwerequan-
titativedetectedbytheanalysisofGC-MS.
MaterialsandMethods
Instrumentation
HS-GC-MS:Alexperimentswereperformedwitha
model7694Eheadspaceautosampler(Hewlet-Pack-
ard, PaloAlto, CA, USA)connectedtoamodelFinni-
gangaschromatograph(TraceMS, USA), interfaced
withamodel5973 massdetectoroperatingintheelec-
tronimpact(EI+)mode.Thegaschromatographwas
equippedwithaPEG-20Mcolumn(30 mlong, 0.25
mmi.d., 0.25 μmfilmthicknes, TraceMSUSA).
RubberseptalinedwithPTFE(150 ℃ proof)were
obtainedfromAnalyticalTechnology(Milan, Italy).
PreparationofT.aurantialbaBroth
T.aurantialbastrain, kindlypresentedbyprofessor
WeijingQuofEastChinaNormalUniversity, andkept
withPDAmediumat28℃, wasusedinthisstudy.The
funguswasgrownin35gsolidseedmediumcontaining
15 gbranand20 mLwater.Solidseedwasincubated
at28 ℃ for7 d.Duringthecultivation, theflaskwas
shakenonceperdayfromthethirdtothefifthday.
Thesolidseedmentionedabovewastheninoculatedin
100 mLliquidseedmediumcontaining2% sucrose,
1% cornpowder, 0.5% peptone.Themedium was
sterilizedunder0.1 MPafor30min.Theseedwasin-
cubatedat27℃inarotaryshaker(ShanghaiPharma-
ceuticalIndustrialAcademe, China)at150 rpmfor2
d.10mLseedculturewasfurtherinoculatedin100 mL
liquidfermentationmedium, andthefermentationwas
carriedoutunderthesameconditionfor7 d.Thefer-
mentationmediumcontained4% sucrose, 0.4% pep-
tone, 1% cornpowder, 1% bran, 0.15% KH2PO4 , and
0.075% MgSO4.
AnalysisofGC-MS
10 mLbrothwastransferredintoglas“headspace”
vials(15 mLvolume), whichwereclosedwith20 mm
rubberPTFElinedsepta(150 ℃ proof)andcrimped
withperforatedaluminiumseals.After40 minequili-
brationtimeonarotatingshakerat50 ℃, samples
wereplacedintheauto-samplerwheretheywereheat-
edat250 ℃ andshakenfor3 minbeforethehead
spacewaswithdrawn.Theheadspaceauto-samplerwas
providedwitha1mLloopkeptat180℃;thetransfer
line, heatedat200 ℃, wasconnectedwiththeinjector
oftheGC(split1:20)viaaninterfaceheatedat250
℃.Theoventemperaturewaskeptat36 ℃ duringthe
injection(3min), thenthetemperaturewasincreased
at4 ℃ /minupto120℃ andthento230℃ atarate
of10℃ minandkeptat230℃ for8min.Heliumwas
usedasthecariergasattheconstantflowrateof0.8
mL/min.
Volatileflavourcompoundsweretentativelyidentified
bymatchingmassspectraldataofsamplecomponents
withthoseofknowncompoundsinadatabase(Nistda-
tabaseandWileydatabase).Folowingequationswere
usedtodeterminetheirpercentageconcentrationofthe
identifiedcompounds(w/w).
Ci=Ai∑Ai×100%
WhereCisconcentrationofonecompound;Aispeak
areacounts;∑Aissummationofalpeakareacounts;
subscriptsIrepresentsonecomponent
ResultsandDiscussion
ThemainvolatileingredientsinT.aurantialba
Broth
Fig.1 Totalionchromatogramforthevolatilefla-
vourinT.aurantialbaBroth
Onlysomeofthemajorpeaksarelabeledinordertomaintainthe
clarityofthechromatogram.
ThePEG-20Mcolumnwasapparentlysuitableforiden-
tifyingandseparatingthevolatileflavorcompounds, as
823Vol.19 ZHANGZhi-caietal:AnalysisoftheVolatileCompoundsinTremelaaurantialbaBrothbyGC-MSMethod
thesharpandnon-overlappingtotalionpeakscouldbe
obtainedasshowninFig.1.Table1 showedthesum-
marizedGC-MSdataandtherelevantpercentageof
eachindividualflavorcompoundwithrespecttotheto-
talvolatileflavorscontents.Morethan94diferentvola-
tileflavorcompoundsinthesubmerged-culturedbroth
weredetected.Amongthem, 34kindsofvolatileflavour
compoundswhosematchindexwasmorethan800were
sesquiterpenes, withtheirtotalcontentpercentagea-
bove47%.Inparticular, thecontentpercentageofRos-
ifoliolwasatanextremelyhighlevelof9.37%, with
theircharacteristicretentionpeaksat27.35and29.28
mininthiscase.
Table1 VolatileflavourcompositionofT.aurantialbaBroth
taR Area Concentrationb MWc Molecularformula Tentativeidentityd
3.79 102500354.4 5.02 C4H8O2 88 aceticacidethylester
4.73 3981099.93 0.19 C2H6O 46 ethanol
4.88 86338944.75 4.23 C4H10O 74 isobutylalcohol
9.97 82543246.76 4.04 C10H16 136 1.6-octodiene, 7-methyl-3-methylene
11.86 4198862.69 0.21 C10H30O5Si5 370 Decamethylclopentasiloxane
13.42 217207174.2 10.63 C5H12O 88 isobutylcarbinol
13.98 12814058.04 0.63 C8H16O 126 3-octanone
14.40 7800087.72 0.38 C5H10O 86 isobutylcarbinol
16.77 34598912.31 1.69 C15H24 204 gurjunene
16.89 3604663.48 0.18 C15H24 204
17.21 3803092.26 0.19 C15H24 204 calarene
17.39 16816984.42 0.82 C15H24 204 4.10.10-trimethyl-tricyclo-(4.4.0.2(1.4))dec-6-ene
17.49 23864818.05 1.17 C15H24 204 athujopsene
17.85 12814739.69 0.63 C15H24 204 guaiene
18.23 11203367.81 0.55 C15H24 204 longifolene
18.28 14804171.09 0.72 C15H24 204 3.4-di(T-butyl)-tolene
18.60 74410001.70 3.64 C15H24 204 neoclovene
18.99 35215922.49 1.72 C15H24 204 longifolene
19.03 14753077.03 0.72 C15H24 204 [ +-] -chamigrene
19.17 7005668.29 0.34
19.23 6290766.30 0.31
19.57 12838811.82 0.63
19.66 7468466.61 0.37
19.77 19886015.32 0.97 C15H24 204 4, 10, 10-trimethy-(-tricyclo-(4, 4, 0, 2(1, 4))dec-6-ene
19.85 44813147.58 2.19 C15H24 204 caryophylene
20.39 23640089.55 1.16 C15H24 204 berkheyaradulene
20.49 3854503.67 0.19
20.61 41290968.22 2.02 C15H24 204 naphthalene
20.76 17068372.61 0.84 C15H24 204 (+)-guaiene
21.09 12419654.55 0.61 C15H24 204 longifolene
21.19 3726497.34 0.18
21.36 3829616.91 0.19
21.51 12368215.20 0.61
21.63 18188222.99 0.89
21.85 47279906.46 2.31 C9H20O 144 elemene
21.97 3899641.80 0.19
22.27 5604355.16 0.27
824 NatProdResDev Vol.19
22.44 8484024.54 0.42 C15H24 204 nonylalcohol
22.57 46282649.59 2.27 C8H8O2 136 benzoicacid, methylester
22.70 7947899.95 0.39
23.00 8012657.03 0.39
23.19 29158322.31 1.43
23.27 8647412.32 0.42
23.37 25072905.47 1.23 C15H24 204 1.6.10-dodecatriene, 7, 11-dimethyl-3-methylene
23.44 42534301.74 2.08 C15H24 204 humalene
23.57 20252223.94 0.99 C15H24 204 2-isopropenyl-4a, 8-dimethyl-1.2.3.4.4a.5.6.7-octahyudro
23.65 4416782.03 0.22 C15H24 204 azalene
23.81 6538532.01 0.32
23.95 5109950.58 0.25
24.20 6684227.47 0.33 C15H24 204 eremophila-1(10), 11-diene
24.35 41583818.09 2.04 C15H24 204 selinene
24.43 77814495.28 3.81 C15H24 204 selinene
24.96 28822411.16 1.41 C15H24 204 delta-cadinene
25.05 26601402.81 1.30 C13H26O0 196 oxime, ethyoxy-phenyl
25.15 5996439.78 0.29 C15 H22 202 tetrahydroionol
25.20 7332189.03 0.36 C15H24 204 1-[ 1, 5-dimethyl-4-hexenyl] -4-methyl-bene,
25.32 10933251.25 0.54 C15H22 202 eudesma-3, 7 [ 11] -diene
26.09 12860175.43 0.63 1s, C13-calamene
26.16 4884136.04 0.24 C17H28 232 5-phenyl-Undecane
26.30 4087421.84 0.20
26.38 5260720.05 0.26
26.60 4648196.04 0.23
26.75 3672979.06 0.18
26.82 11298732.34 0.55
27.03 8361586.20 0.41
27.18 40363979.19 1.98 C8H10O 122 phenylethyalcohol
27.28 6790902.66 0.33
27.35 148634559.85 7.28 C15H24O 220 rosifoliol
27.50 6008762.15 0.29
27.55 3914275.25 0.19
27.70 4198840.02 0.21
27.86 3888206.37 0.19
28.21 11001799.76 0.54
28.26 13449855.44 0.66
28.42 42658349.10 2.09 C15H24O 220 rosifoliol
28.62 10533455.54 0.52 C15 H26O 222 ledol
29.10 7068164.06 0.35
29.28 110502232.06 5.41 C9H10O3 166 benzoicacid, 4-methoxy, methylester
29.71 15449561.48 0.76 C10H12O3 180 benzoicacid, 4-methoxy, ethylester
30.26 4063818.71 0.20
30.62 22285124.79 1.09 C15 H18 138 azulene, 1.4-dimethyl-7-(1-methyethyl-)
30.86 5393379.21 0.26 C15H22O 218 4.6.6trimethyl-2-(3-methylbuta-1, 3-dienyl)-3-oxotricyclo[ 5.1.0(2, 4)octane
30.90 3865887.07 0.19
30.96 12406973.07 0.61 C15H22O 218 2(11, 1)naphthalenone, 3, 5, 6, 7, 8, 8a-hexohydro-4, 8a-dime-thyl-6-[ 1-methlethenyl]
31.05 4938914.48 0.24 C14H22O 206 4-[ 2.2-dimethyl-6-methylenecyclohexylidene] -3-methylidene-2-one
825Vol.19 ZHANGZhi-caietal:AnalysisoftheVolatileCompoundsinTremelaaurantialbaBrothbyGC-MSMethod
31.34 5355641.72 0.26
33.39 4965227.23 0.24
34.47 17894503.18 0.88 C9H9BrO2 227 5′-bromo-2′-methoxyacetophenone
35.66 3405072.66 0.17
36.19 5503314.91 0.27
aretentiontime;btheconcentrationofonecompoundisrepresenteditsarea%;cmolecularweight;dthecompoundswerenotgiveninthetablewhosematch
indexwaslessthan800.
PharmaceuticaleffectofT.aurantialba
Theoxygencontainingderivativeofsesquiterpeneshas
strongaromaandbioactivity.Theyexistintheformsof
melow, ketone, lactone, glycoside(hawksworth1995),
orasalkaloidinplants.Becausesesquiterpeneshave
manydiferentandimportantbiologicalfunctionsand
potentialforegroundofapplication, ithasbeenbecom-
ingmoreandmoreatractive.Themajorsourcesfor
producingsesquiterpenesreliesonthenaturalplants
resources.However, naturalplantresourcesforprodu-
cingsesquiterpeneswereverylimited.Asaresult, to
findanalternativesourceofproducingsesquiterpenes
isofthemostinterests.Thisstudyshowedthatsesquit-
erpeneswerethemajorvolatileingredientsofthesub-
merged-cultivationofT.aurantialba.Furtherandcom-
prehensiveinvestigationisscheduledtobeimplemen-
tedinthenextresearchstage.
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