中南大學(xué)中藥現(xiàn)代化研究中心

ChemometricsandMultivariateResolutionanditsApplicationinanalysisofTCM中南大學(xué)中藥現(xiàn)代化研究中心梁逸曾Chemometrics

Chemometricsisanewchemicaldisciplinethatusesthetheoryandmethodsfrommathematics,statistics,computerscienceandotherrelateddisciplinestooptimizetheprocedureofchemicalmeasurement,andtoextractchemicalinformationasmuchaspossiblefromchemicaldata.Chemometricscouldbedefinedasadisciplineoffundamentaltheoryandmethodologyofchemicalmeasuring.化學(xué)計量學(xué)運用數(shù)學(xué)、統(tǒng)計學(xué)、計算機科學(xué)、以及其他相關(guān)學(xué)科的理論與方法，優(yōu)化化學(xué)量測過程，并從化學(xué)量測數(shù)據(jù)中最大限度地獲取有用的化學(xué)信息，可以說是一門化學(xué)量測的基礎(chǔ)理論與方法學(xué)。BriefHistoryofChemometricsSvanteWolduseditfirstlyforapplyingscientificprojectin1970inSweden;WoldandKowalskifoundedtheInternationalChemometricSocietyin1974;AnalyticalChemistrypublishedspecialreviewon“Chemometrics”everytwoyearsince1978;Twospecialchemometricinternationaljournalsnamed“J.Chemom.”and“ChemLab”appearedin1987frombothAmericanandEurope.ContentsinchemometricsChemometricsconsistsoffundamentalandmethodologyofchemicalmeasurements.NecessaryfundamentalknowledgeofstatisticsandlinearalgebraVectorandMatrixIsMathematicsreallyusefulforchemists?Dataexploding;Extractchemicalinformationfromthedata;RevolutionofInformationtechnique;ProgressinComputerability;VectorinanalyticalchemistryAllthespectra,chromatogramsandetc.canbenumeratedintoagroupofnumbers,whichiscalledavectorinmathematics.

HyphenatedInstruments,suchasHPLC-DAD,GC-MS,GC-IR,HPLC-MSTwo-waydatacontainingbothchromatographyandspectra;Datamatrixwithmorethan10Megabytes;DatabaseoflotsofchemicalstandardsThemixturespectrumoftwodifferentchemicalcompoundsaandbaccordingtotheLambert-BeerlawGeometricsenseofvectoradditionGeometricsenseofvectorsubtractionDirectionandlengthofvectorThedirectionofavectorisdecidedbyallitselements,sincethedifferentratiosbetweenthemcandefinesdifferentdirectionsinlinearsubspace;Thelengthofavectorisalsodecidedbymagnitudesofitselements,sincewehave,||a||=(a12+….+an2)1/2differentratiosbetweenthemcandefinesdifferentdirectionsinlinearsubspaceSubtractionoftwovectorsdefinesthedistancebetweenthetwopointsinndimensionalspaceNumericalmultiplicationofvectorsThespectraofdifferentconcentrationsInnerproductandouterproductbetweenthevectors

InnerordotproductbetweentwovectorsproducinganumberGeometricsenseofinnerproductbetweentwovectorsInnerproductandprojectionbetweenvectorsOuterproductbetweentwovectorsproducingabilinearmatrix,whichisofspecialimportanceinmultivariateresolutionWhatisthechemicalmeaningofrankofamatrix?Linearlyindependent?Rankofamatrix=thenumberofthechemicalcomponentsinthemixture?Mixturenumberandcompoundnumber?Apartofarealtwo-waydatameasuredfor

CortexCinnamomi(肉桂)

Lambert-BeerLawTheproblemhereforchemiststosolveisthat,withthemeasurementmatrixathand,oneneedstofindout:Thenumberofabsorbingchemicalcomponents:AThespectrumofeachchemicalcomponent:si(i=1,2,…,A)(firststepof

qualifications)Theconcentrationprofileofeachchemicalcomponent:ci(i=1,2,…,A)(firststepfor

quantification)Isitpossible?Yes,ifwehavethetwo-waydata!!MixturespectraandcompoundspectraThisisatwocompoundsystem;Tworedvectorsarethecompoundspectra;Sevenbluevectorsaremixturespectra;

LinearlyindependentThetwospectraarefromtwodifferentchemicalcompounds,sotheyareindependentwitheachotherSevenspectraareallfromthespectraofthetwochemicalcompounds,sotheyaredependentupontheconcentrationsofthetwocompoundsinthemixtureMixturenumberandcompoundnumber

Rankofamatrix=thenumberofthechemicalcomponentsinthemixture?

Unchangedarechemicalcompounds;Rankofamatrixisalsocertain,whichisnotchanged;Thus,ifwecollectallthespectratoformamatrix,therankofwhichshouldbe2.Howcanwefindtherankofamatrixwithmeasurementnoise?Whatarenoisesandhowdotheyinfluencethedataanalysis?Thus,weneedstatisticsandalsoalgebra;Usefultechnique:Principalcomponentanalysis!!Thisisreallyadifficultproblem!Now,let’sgothroughthisproblemstepbystep.SinglevaluedecompositionQuestion2Wehaveadatamatrixathand,weneedtoknowtherearehowmanychemicalcomponentsinit.Whattechniquedoyouwanttouse?Principalcomponentanalysis?Canweusesingularvaluedecompositiontosolvetheproblem?MultivariatecalibrationandmultivariateresolutionThisisthemajortopicinanalyticalchemometrics,wewilldiscussitinsomedetail.SinglevaluedecompositionandnumberofchemicalcomponentinthesystemLocalfactoranalysisandZerocomponentregionsEvolvingfactoranalysis(EFA)Fix-sizedmovingwindowevolvingfactoranalysis(FSMWEFA);Eigenstructuretrackinganalysis(ETA)EvolvinglatentprojectionsEvolvingFactorAnalysisEvolvingfactoranalysisFix-sizedMovingWindowEvolvingFactorAnalysis(FSMWEFA)Fix-sizedMovingWindowEvolvingFactorAnalysis(FSMWEFA)SomeproblemswithlocalfactoranalysisLocalrankZerocomponentregions,WindowsizeEigenstructuretrackinganalysisRankmapandlocaldatastructureEvolvinglatentprojectionsPrincipalComponentanalysisandLatentprojectionsEvolvinglatentprojectionsSelectiveinformationandEvolvinglatentprojections

ZerocomponentregionandnoiselevelAsimpleexampleResolutionintopurechromatogramsandspectraBecauseA=CStIfwehavepurespectraS,thenwecanusethefollowingequation:

C=AS(StS)-1Resolutionintopurechromatogramsandspectra美國標準局16組分PAH混合物標樣(Sulpeco)

已知峰9為苯并[a]蒽和屈，峰14為苯并芘和二苯并蒽的二組分重疊峰Peaks5and6intheplot峰5、峰6的演進特征投影圖

峰5的前5個特征值依次為16382，2436，1294，22，11分辨所得的芴、苊、菲、蒽的色譜與光譜J.CGiddingsin“Statisticaltheoryofcomponentoverlapinmulticomponentchromatograms”claimedthatItisshownthat,relativetothemaximumpeakcontentorpeakcapacityforcloselyspacedpeaks,arandomchromatogramwillnevercontainmorethanabout37%ofitspotentialpeaksand,worstofallfromananalyticalpointofview,18%ofitspotentialsingle-componentpeaks.Thenumberofobservedpeaksisnot,then,thesameasthenumberofdistinctchemicalcomponents.Thelossofanalyticalinformationresultingfromthisoverlapisbyitselfserious,buttheseverityoftheproblemisgreatlymagnifiedifwedonothaveagoodestimateofthemagnitudeoftheloss.

Anal.Chem.1983,55,418-424

ApplicationtoenvironmentalchemistryPAHsrepresentaclassoforganicpollutantsthatarewidelyfoundintheenvironment.Theyhavebeenshowntoexertaverystrongcarcinogenicactiononanimals.ThisisanexampleforqualitativeandquantitativeanalysisofPAHsinair-borneparticulatesinHongKongcitybythechemometricresolutionmethod.

1.8.2

HELP分辨

直觀推導(dǎo)式演進特征投影法充分利用了二維數(shù)據(jù)的色譜信息和波譜化學(xué)信息，以逐步推導(dǎo)的方式分辨各組分的純色譜和波譜。具體步驟簡述如下：

1）將響應(yīng)矩陣X沿色譜方向分解為一系列相對簡單的子體系Xi，檢測各個子體系Xi存在的背景與噪聲，扣除背景及噪聲

2）利用演進特征投影圖和特征結(jié)構(gòu)跟蹤分析與秩圖確定其組分數(shù)以及各組分的選擇性區(qū)域和零濃度區(qū)域

3）利用各組分的選擇性區(qū)域和零濃度區(qū)域分辨出各組分的純色譜和波譜

4）將解析所得的各組分的純色譜和波譜乘積線性加和與實際數(shù)據(jù)比較，驗證解析結(jié)果的可靠性

Figure2.6’TheamplifiedtotalioncurrentchromatogramoffractionN6obtainedfromGC-MS.

1.9定性分析

用質(zhì)譜庫相似檢索定性，結(jié)合色譜保留指數(shù)確正質(zhì)譜的定性結(jié)果

色譜峰B分辨所得六種物質(zhì)定性，它們依次是：6-methoxy-2-methyl-beta-carboline(C13H12N2O)、3,3’-dimethyl-[1,1’-biphenyl]-4,4’-diamine(C14H16N2)、2,5-dimethyl-4-[3-amino-4-methylphenyl]pyridine(C14H16N2)5-acetyl-4-amino-2-methylthio-thiophene(C8H8N2OS2)、4,9-dimethyl-naphtho-[2,3-b]-thiophene(C14H12S)、1,2,5,6-tetramethylacenaphthylene(C16H16)

依此原理，分辨七個餾分的所有色譜峰，對各個組分進行定性分析。在1200多組分中鑒別出130多種含氮化合物

化學(xué)計量學(xué)與中藥現(xiàn)代化研究中草藥的現(xiàn)代化與國際化是勢不可擋的世界潮流美國自30到60年代，幾乎停止植物藥的研究（Interestinmedicalplantsprogressingsignificantlydiminishedinthe1930sandbotancalsfellintoalmostcompletedisuseuntil1960s,RoyUpton,HerbalistExecutiveDirector,AmericanHerbalPharmacopoeia），到現(xiàn)在（2000）

GlobalHerbalMarket:US$19.6billion!!（196個億美金）；德國和法國聯(lián)合開發(fā)銀杏葉提取物EG6761，找到最佳銀杏提取物的指紋圖譜，導(dǎo)致植物藥的在歐美重新流行HealthProducts(WesternCountries)GinggoBiloba

(銀杏)St.Jone’sWort(貫頁連翹)HealthProducts(WesternCountries)Greentea(綠茶)Echinacea(紫錐花)TraditionalChineseMedicine(TCM)HealthProductsDongQuai(當歸)Ginseng(人參)TraditionalChineseMedicine(TCM)HealthProductsCordyceps(冬蟲夏草)Galic(大蒜)NaturalProductMarketGlobalHerbalMarket2000:US$19.6billion!!

NaturalProductMarket

缺乏適當或可接受的評價傳統(tǒng)草藥的研究方法Despiteitsexistenceandcontinueduseovermanycenturies,anditspopularityandextensiveuseduringthelastdecade,traditionalmedicinehasnotbeenofficiallyrecognizedinmostcountries.…

Thequantityandqualityofthesafetyandefficacydataontraditionalmedicinearefarfromsufficienttomeetthecriterianeededtosupportitsuseworldwide.

Thereasonsforthelackofresearchdataareduenotonlytohealthcarepolicies,

butalsotoalackofadequateoracceptedresearchmethodologyforevaluatingtraditionalmedicine.傳統(tǒng)植物化學(xué)方法研究中藥的窘境每個化學(xué)物質(zhì)都分離了（耗時），但藥效似乎都不強，找到活性單體藥無法與同類西藥相比；中藥的君臣佐使，協(xié)同作用難以體現(xiàn)；日本人、德國人（銀杏）的發(fā)現(xiàn)，指紋圖譜的提出；中藥現(xiàn)代化需要新思路，新方法。中草藥現(xiàn)代化與國際化研究中的幾個難點問題中草藥的化學(xué)基礎(chǔ);(多組分復(fù)雜混合體系)中藥復(fù)方制劑;(“膏丹丸散,神仙難辯”);中藥國際化的主要難點：中藥基地軌范化(GAP和GEP)，中藥生產(chǎn)標準化(GMP)，基于化學(xué)指紋圖譜的中藥質(zhì)量控制;中藥作用機制研究（君臣佐使,協(xié)同作用）中藥藥效及藥代動力學(xué)方法的建立;中藥譜（化學(xué)指紋圖譜）效（藥理藥效）學(xué)的創(chuàng)立;中醫(yī)藥理論科學(xué)化的艱巨任務(wù)廣西玉林廣東肇慶越南云南中藥指紋圖譜在中藥現(xiàn)代化研究中的核心地位EssentialConstituentsofRamulusCinnamomi(桂枝）

中藥現(xiàn)代化的機遇與挑戰(zhàn)基于中藥指紋圖譜的質(zhì)量控制、基于中藥指紋圖譜的藥效研究、基于中藥指紋圖譜的藥代動力學(xué)研究、餾分組合（雞尾酒方法），新型更有效復(fù)方制劑的發(fā)現(xiàn)與開發(fā)、全依賴于中藥指紋圖譜的快速定性定量解析！中藥色譜指紋圖譜的測定方法以解決中藥長期處于“丸散膏丹，神仙難辨”的被動局面InstrumentsCommonlyusedforChemicalAnalysisofChineseMedicineThinLayerChromatography(TLC)(薄層色譜)

qualitativeanalysis&semi-quantitativeanalysisHighPerformanceLiquidChromatography(HPLC)(高效液相色譜)

Bothqualitativeanalysis&quantitativeanalysisGasChromatography(GC)(氣相色譜) Bothqualitativeanalysis&quantitativeanalysisDNAAnalysis(DNA分析)CapillaryElectrophoresis(CE)(毛細管電泳)中藥指紋圖譜剖析的新工具聯(lián)用色譜儀器HyphenatedInstrument(聯(lián)用儀器)Mostmodernandadvancedanalyticalinstrument.

e.g.HPLC-DAD,GC-MS,CE-DAD,etc.Combinationoftwoormoreinstrumentse.g. HPLC DAD(diode-arraydetector)

GC MS(Mass-spectrometer)SeparationInstrumentSignal

InstrumentGetmoredata/informationHPLC-DAD3DchromatogramHPLCchromatogramofnucluesideofCordycepsSinensis(冬蟲草)atonewavelengthHyphenatedInstrument(聯(lián)用儀器)HyphenatedInstrument(聯(lián)用儀器)GCinstrumentGCchromatogramof

pepticpowder(平胃散)Massspectrumtakenatretentiontime10.2minutesHyphenatedInstrument(聯(lián)用儀器)Advantages:Moredataisobtainedataretentiontime.Morespectralinformationacquiredtogivethreedimensional(3D)data.MuchmoreinformationavailabletoanalyzecomplicatedsystemlikeTCM.HyphenatedInstrument(聯(lián)用儀器)Disadvantages:Hugeamountof3Ddataisobtained.Needdataprocessingmethodsforinformationextraction,patternrecognition,etc.Computerpowerwaspoorbefore.Usualpractice:UseafewdataobtainedfromHPLC-DAD/GC-MSto findmarkercomponentsoractiveingredients.

Now,everythingbecomespossiblewithhightech.!

國家藥品監(jiān)督管理局付局長任德權(quán)先生在現(xiàn)代化中藥產(chǎn)業(yè)關(guān)鍵技術(shù)系列研討會的發(fā)言中指出：

實際上，從分析化學(xué)學(xué)科自身發(fā)展看，純品的分析現(xiàn)已無特殊困難，而對復(fù)雜有機混合物體系給出快速定性定量分析，...。從現(xiàn)代化學(xué)計量學(xué)的觀點看，儀器分析數(shù)據(jù)的傳統(tǒng)處理方法“如光譜、波譜只取其峰值，對色譜只計算其面積等，這樣做的后果只能造成大量有用信息的浪費?！泵鎸π涡紊亩嘟M分體系，在充分利用化學(xué)量測儀器所產(chǎn)生的化學(xué)信號特點的基礎(chǔ)上，化學(xué)計量學(xué)運用統(tǒng)計學(xué)和應(yīng)用數(shù)學(xué)及計算機，最大限度地從中抽取不同樣本中的定性、定量化學(xué)信息。在這方面我國學(xué)者梁逸曾先生按對被分析的多組分體系的定性組成知悉程度，提出了白色、灰色和黑色多組分體系三種分析體系。現(xiàn)代分析化學(xué)學(xué)科，特別是化學(xué)計量學(xué)的發(fā)展，為中成藥指紋圖譜用于定性定量分析，提供了現(xiàn)代科學(xué)技術(shù)基礎(chǔ)?！瓪v史的挑戰(zhàn)與機遇并存,我們則無旁貸,充分利用時代的條件,去解決時代的問題.開創(chuàng)打破“神仙難辯”,逐步實現(xiàn)中藥定性定量分析的新時代.中藥指紋圖譜的定性定量剖析聯(lián)用色譜與化學(xué)計量學(xué)TraditionalChinesemedicines我們對二十幾種單味藥和復(fù)方制劑進行了HPLC-DAD和GC-MS分析;對某些單味藥的道地性、季節(jié)性影響也進行了系統(tǒng)研究，以得到它們的mono-graphs；對我省正清制藥集團的魚腥草和風(fēng)痛寧注射液指紋圖譜的建立進行了系統(tǒng)的方法學(xué)研究。Chemometrics:CordycepssinensisChromatogramsofthefungalpartofCordycepssinensisatwavelength(a)234nm,(b)260nmandthelarvapartofCordycepssinensisatwavelength(c)234nm,(d)260nm.AsimpleexampleChemometrics:Cordycepssinensis(a)Theselectivechromatogramsofthefunguspartfrom11.0to14.0minuteswithanintervalof5nmand(b)showsthespectraofthefunguspartintherangeof195to312nmwithanintervalof0.1minute.(c)and(d)showthecorrespondingeigenvaluesplotandthelatentprojectgraphof.(e)and(f)showtheresolvedchromatogramsandspectraforcomponentsf1andf2inthefungalpart.Chemometrics:CordycepssinensisTheHELPmethodwasusedforresolvingthesechromatograms.TheoverallresolvedchromatogramofthefungalpartTheoverallresolvedchromatogramofthelarvalpartFanGong,Yizengliang,F.t.Chau,Anal.Lett.,33（2000）2105-2128GanFengandYizengLiang,AnalyticalScience,16(2000)603-607ChengjianXu,YizengLiangandJianhuiJiang,AnalyticalLetters,33(2000)2105-2128HailinShen,YizengLiang,O.K.KvalheimandR.Manne,ChemometricsandIntelligentLaboratorySystems,51(2000)49-59(inEnglish)HailinShen,L.Stordrange,R.Manne,O.M.KvalheimandYizengLiang,ChemometricsandIntelligentLaboratorySystems,51(2000)37-47(inEnglish)HailinShen,XiaoningLi,YizengLiang,ChineseScienceBulltin（科學(xué)通報）,45(2000)587-592HailinShen,Youqunsong,HuiCui,YizengLiang,ActaChimicaSinica（化學(xué)學(xué)報）,58(2000)438-442QingsongXu,YizengLiangandKaitaiFang,ChemometricsandIntelligentLaboratorySystems,52(2000)155-166.Pepticpowder PepticpowderisanancientconcentratedpreparationinChina.

Itcanclearawaypathogenicdampness,moisturizespleen,promotethecirculationofqiandregulatethestomach.Theformulationiscomposedoffoursingleherbs.TheyareRhizomaAtractylodis(atractylodesrhizome),PericarpiumCitriReticulatae(tangerinepeel),CortexMagnoliaeOfficinalis(magnoliabark)andRadixGlycyrrhizae(licoriceroot).Thevolatileconstituentsofpepticpowderarepharmacologicalactive.GongFan,Y.Z.Liang,HuiCui,F.T.Chau,BennyT.P.Chan,Determinationofvolatilecomponentsinpepticpowderbygaschromatography-massspectrometryandchemometricresolution,J.ChromatographyA,905（2001）193-205(inEnglish)GongFan,Y.Z.Liang,Qing-SongXu,F.T.Chau,Gaschromatography-massspectrometryandchemometricresolutionappliedtodeterminationofessentialoilsinCortexCinamomi,J.ChromatographyA,909（2001）237-247(inEnglish)龔范,梁逸曾,宋又群，彭源貴,崔卉,A.K.M.Leung,Foo-timChau，高等學(xué)校化學(xué)學(xué)報,2001(inpress).GanFeng,Jia-hongYangandY.Z.Liang,Liberarysearchofmassspectrawithanewmatchingalgorithmbasedonsubstructuresimilarity,AnalyticalScience,17(2001)635-638(inEnglish)Y.Z.LiangandO.M.Kvalheim,Resolutionoftwo-waydata:theoreticalbackgroundandpracticalproblemsolvingPart1:theoreticalbackgroundandmethodology,FreneniusAnal.Chem.,370(2001)694-704.(inEnglish)MinghaoZhangandYizengLiang,Analyst,

(2001)(inpress).GongFan,Y.Z.Liang,Qing-songXu，F(xiàn).T.Chau,King-manNg,Anal.Chim.Acta,2001(inpress).(inEnglish)Y.Z.Liang,K.T.Fang,Q.S.Xu,ChemometricsandIntelligentLaboratorySystems,2001(inpress).

ChengjianXu,Y.Z.Liang,You-QunSongandJi-shanLi,FreneniusAnal.Chem.,(2001)(inpress)

fingerprintsforsomeherbalmedicinestToppart:(蒼術(shù)揮發(fā)油);Lowerpart:(厚樸揮發(fā)油)FindingFingerprint:HELPGC-MSofpepticpowder(平胃散)pepticpowder(平胃散)Therearefourherbsinpepticpowder:sayRhizomaAtractylodis(atractylodesrhizome),PericarpiumCitriReticulatae(tangerinepeel),CortexMagnoliaeOfficinalis(magnoliabark),andRadixGlycyrrhizae(licoriceroot).Theresultsobtainedsofarshowedthatthevolatileoilinpepticpowderarealmostthemixtureoftheabovefourherbsproportionaltotheirrelativeamountstakeninthepreparation.

FindingFingerprint:

ChemicalCompositionApproachATCMpreparation:Si-wudecoction(四物湯)100gRadixAngelicaeSinensis,當歸80gRhizomaChuanxiong,川芎120gRadixPaeoniaeAlba,白芍

120gRadixRehmanniaePraeparata,熟地VolatileoilextractionAccordingtothestandardextractionmethodinChinesePharmacopoeiaFindingFingerprint:

ChemicalCompositionApproachRhizomaChuanxiong川芎RadixAngelicaeSinensis當歸FindingFingerprint:

ChemicalCompositionApproachPreliminaryresults

ComponentsfoundComponentsidentifiedRelativecontentRadixAngelicaeSinensis當歸805092.09%RhizomaChuanxiong

川芎1278284.09%Si-wudecoction

四物湯975785.27%當歸、川芎與復(fù)方四物湯揮發(fā)油化學(xué)組分的定性定量結(jié)果鑒定了當歸、川芎及四物湯揮發(fā)油的50、82和57個化學(xué)組分，約占總量的92.02%、84.09%和85.27%。結(jié)果表明,在16個具有藥理活性的主要化學(xué)組分中，有6個組分含量明顯增加，4個組分基本不變，1個組分減少，另有5個組分在四物湯揮發(fā)油中檢測不到。反式藁本內(nèi)酯的相對含量增加得很多，而它正是四物湯復(fù)方中抗膽堿、抑制子宮收縮最強的組分；丁基酞內(nèi)酯、丁烯基酞內(nèi)酯相對含量也是成倍增加，這2個組分均是復(fù)方具有解痙作用的主要成分，同時兼有抑制子宮收縮與平喘的作用；4-松油醇、

-松油醇、香茅醇具有平喘、消毒與抑制病菌的功能，其相對含量比理論計算值也略有增加。為詳盡地研究由單味藥配伍成復(fù)方四物湯時各化學(xué)成分在質(zhì)和量上的變化，以更好地說明該復(fù)方配伍的科學(xué)性和各單味藥之間的協(xié)同作用，我們將對復(fù)方與各單味藥提取揮發(fā)油后的水相部分進行更進一步的定性定量檢測。利用化學(xué)計量學(xué)方法分析遼五味子揮發(fā)性成份的GC/MS數(shù)據(jù)遼五味子是木蘭科植物五味子的干燥成熟果實，主產(chǎn)我國東北，認為有益氣、明目、補不足、養(yǎng)五臟、壯筋骨等作用，中醫(yī)作為收斂、滋補強壯劑。遼五味子粉碎后有愉快的香氣。

六種不同提取方法得到樣品的TIC圖兩個來自不同樣品的