1、超臨界流體萃取,層析及其他應用Supercritical Fluid Extraction,Chromatography and Other Applications林華經引光生物科技有限公司第1页，共50页。What is a Supercritical Fluid?When the pressure and temperature of a substance is raised above its critical pressure and critical temperature(the critical point)the substance enters the supercrit
2、ical state.A Supercritical Fluid is a substance with both gas-and liquid-like properties.第2页，共50页。Critical Temperature and PressurenThe Critical Temperature(Tc)is the maximum temperature at which a gas can be converted to a liquid by increasing the pressure.nThe Critical Pressure(Pc)is the maximum p
3、ressure at which a liquid can be converted to a gas by increasing the temperature.第3页，共50页。Phase Diagram of Carbon DioxideTemperatureSupercriticalFluidSolidLiquidTriple PointCritical Point31.3 oC,1072 psiPressureGas第4页，共50页。Physical Properties of CO2PhaseGasSCFLiquidDensity0.62.0 x 10-30.2 0.90.8 1.
4、0viscosity0.53.5 x 10-42.09.9 x 10-40.32.4 x 10-2Diffusivity0.01 1.00.53.3 x 10-40.52.0 x 10-5第5页，共50页。Advantages of Supercritical Fluids as solventsnSolvating power related to density(at constant T)nGas-like mass transport propertiesnFacile penetration into porous material第6页，共50页。Critical Paramete
5、rs of Common FluidsTc(C)Pc(atm)c(g mL-1)CO231.372.90.47N2O36.572.50.45SF245.537.10.74NH3132.5112.50.24H2O3742270.34n-C4H1015237.50.23n-C6H1219733.30.23Xe16.658.41.10CCl2F211240.70.56CHF325.946.90.52Source:“SFE and Its use in Chromatographic Sample Preparation”Ed.S.Westwood.Chapter 1第7页，共50页。Advantag
6、es of CO2 for SFEnLow critical parametersnInert,Non-toxic,Nonflammable,Non-corrosivenEasily purified(inexpensive)nNonpolar:dielectric constant similar to hexanenModifiers can be used to increase polarity第8页，共50页。Supercritical Fluid Extraction(SFE)Basic Theory第9页，共50页。RestrictorCO2sample cellin heate
7、dchamberCO2TrapSolid or LiquidModifier PumpSFE System ComponentsCO2 Pump(high pressure)Liquid Carbon Dioxide(requires a dip tube)第10页，共50页。Advantages of SFE over Solvent ExtractionnFasterlResults in minutes rather than hoursnSelectivity results in cleaner extractsnLow Critical ParameterslHandling of
8、 thermally labile analytesnNon-hazardous solventsnAutomatednCost per test is lowernLimited or no solvent removal requirednNo solvent disposal costs第11页，共50页。The Mechanism of SFEA Three Step ProcessDissolving/Resolving analyte(s)2.Sweeping the analyte(s)from the matrix3.Trapping the analyte(s)第12页，共5
9、0页。SFE MechanismSC-CO2+dissolved analyte to the trapHigh PressureLiquidCO2SC-CO2 diffuses matrix,dissolves and resolvesanalyte from the matrixHeat第13页，共50页。TrappingTrapping SolidHigh Surface AreaAdequate AmountTrapping LiquidHigh Surface TensionAnalyte is SolubleLow VolatilityPressurized Cryogenical
10、ly CooledGaseousCO2GaseousCO2CO2 is changingfrom a SF(2 ml/min)to an expanded gas(1 L/min)Analyte no longer solubleMechanical movementof analyte due to therapid expansion requiresthe use of trappingmaterial第14页，共50页。SFE-Method DevelopmentnPressurelIncrease pressure increases density=increase in solu
11、bilizing power.nTemperaturelIncrease temperature may decreases density=decrease in solubilizing power(ie CO2 at 100 bar)nExtraction TimenFlow RatenFluid CompositionlCo-solvents/modifierslReactant AdditivenStatic vs.Dynamic Extraction第15页，共50页。Why Use Modifiers?Analytes that have polar components req
12、uire the use of a co-solventTriglycerides CH2OCOR1R2COOCH CH2OCOR3 CH2OCOR1R2COOCH O CH2OPOR3 O-PhospholipidsR 1&2 groups are long chain hydrocarbons(nonpolar),while R 3 contains phosphorus and nitrogen and is polarR 1,2&3 groups are long chain hydrocarbons(nonpolar)第16页，共50页。Modifiers(Co-solvents)i
13、n SFERole of Modifiers in SFEChanges in Solvent Polarity*Interaction with Matrix*Interaction with AnalyteMethods of Addition*Directly into extraction cell(spiking)*On line modifier addition(uses a second pump)第17页，共50页。SFE vs.Traditional Sample Extraction MethodsConclusionSFE can be versatile,select
14、ive and fasterSFE reduces hazardous solvent use and costSFE can produce cleaner,more concentrated extracts for post extraction analysis第18页，共50页。Analysts Time Allotment第19页，共50页。Routine and Novel Applications of Analytical SFE第20页，共50页。SFE for Research第21页，共50页。Environmental SFE ApplicationsMatrices
15、 Soil Tissue Clay Sandy Loam Sludge River Sediment Marine Sediment Fly Ash Incinerator Ash Target AnalytesTPHPAHPCBsPesticidesDibenzofuransDioxins第22页，共50页。Approved Methods EnvironmentalUS EPA 3560-TPH in Soil:Supercritical Fluid Extraction of Total Recoverable Petroleum Hydrocarbons US EPA 3561-PAH
16、 in Soil:Supercritical Fluid Extraction of Polyaromatic Hydrocarbons US EPA 3562-PCB and OCP:Supercritical Fluid Extraction of Polychlorinated Biphenyls(PCBs)and Organochlorine US EPA 3545:Pressurized Fluid Extraction(PFE)USDOE STD-3013-99:Determination of Residual Water in Impure Plutonium OxidesAO
17、AC draft:SFE-GC/MS determination of pesticide residues in non-fatty fruits and vegetables 第23页，共50页。Pharmaceutical/Natural Product Applications第24页，共50页。SFE of Natural Products-Roger M.Smith LC-GC International,Jan.1996,9-15Catharanthus roseusGerman chamomileMagnolia grandifloraTansyChamomile flower
18、headsGingerPeppermintThymeClove OilKola nutsPimentoTurmericDragon headLavenderPoppy seedsWheat germ oilEnglish yewLemon grassRosemaryFeverfewLemon peelSavory第25页，共50页。Microbial Natural ProductsR.M.Smith,op.cit.OrganismExtractAgaricus speciesCarboxylic and fatty acidsBeuveria niveaCyclosporinFilament
19、ous fungiPolyunsaturated fatty acidsFlour,moldy bread,mushroomsErgosterolMoldy branSterolMoldy grainAflatoxin第26页，共50页。Extracts from BiomassR.M.Smith,op.cit.MicroorganismExtractActinomycete speciesMycolutein and luteoreticulinActinomycete speciesOligomycin AAspergillus fumigatusSydowinin B and epoxi
20、deBipolaris urochloaeOphiobolin APenicillium expansumChaetoglobosin APenicillium sclerotium(+)-SclerotiorinStreptomyces speciesElaiophylin第27页，共50页。Summary R.M.Smith,op.cit.Extracts typically cleaner than those obtained with organic solvents.Mild conditions minimize degradation.SFE methods are faste
21、r than organic solvent extractions.第28页，共50页。Extraction of Pharmaceuticals Using Pressurized Carbon DioxideJ.R.Dean,S.Khundker,J.Pharm.&Biomed.Anal,15(1997)875-886 Recoveries from 81%-95+%CO2 and CO2 with modifiers Generally faster than other methods with better selectivity for target analytes.Preco
22、ncentration steps could be eliminated in some cases.Liquid matrices required immobilization on solid support or SPE cartridge.第29页，共50页。Animal Feeds J.R.Dean,S.Khundker,op.cit.AnalyteMatrixMenadioneRat chowTipradaneRodent dietHypolipidermic drugRat feedHalogenated aromatic phenoxy derivativesDog fee
23、d/rodent feedAtovaquoneRat feedFluconazoleAnimal feedPropanolol,Tamoxifen,ZM 95527,169369Rodent diet第30页，共50页。FormulationsJ.R.Dean,S.Khundker,op.cit.AnalyteMatrixMegesterol AcetateTabletFelodipineTabletBenzodiazipines(7)Tablet/capsuleCaffeine,vanillinTabletVitamin A,ETabletRetinol palmitate,tocopher
24、ol acetateOintmentPolymyxin B sulphateCream/OintmentAcylvoirOintmentSulfamethazole,trimethoprimSeptra infusionTriamincinoloneDermatological patchesMisoprostolHydroxypropyl methylcellulose第31页，共50页。Biological MatricesJ.R.Dean,S.Khundker,op.cit.AnalyteMatrixVeterinary drugs(4)Pig kidneyNitrobenzamide
25、residueLiverCodeine,morphine,ethyl morphineHairKetorolac,flavonePlasmaMebervine alcoholDog plasmaMorphineSerumBeudesonidePlasmaCaffeineKola nutsTaxanesYew tree needlesChinese herbal medicinesPlantsDiosgeninTubers of Dioscorea nipponicaTaxol and baccatin IIINeedles of Taxus cuspidataZingiber zerumet
26、rhizomesPlantsMevinolin and hydroxy acid formFermentation brothPhylloquinoneSoy protein and infant formula第32页，共50页。MiscellaneousJ.R.Dean,S.Khundker,op.cit.AnalyteMatrixTriprolidine,pseudoephedrineAqueousSteroids(10)AqueousIbuprofenAqueous第33页，共50页。Natural Materials StudiedM.J.Noh,et.al.,op.cit.Spec
27、ific NamePart UsedLycium chineseFruitSchizandre chinensisFruitCitrus unshiuFruit barkAngelica gigasRootCornus officinalisFruitCnidium officinaleRhizomeGinko bilobaLeafAralia cordataRootEvodia officinalisFruitCrataegus pinnatifidaFruitPaeonia lactifloraRootLeonurus sibricusAllSophora japonicaFlowerAr
28、temisia capillarisAllPlatago asiaticaSeed第34页，共50页。Natural Materials Studied,contd.M.J.Noh,et.al.,op.cit.Specific NamePart UsedEphedra sinicaAllAconitum carmichaeliTuberScolopendra subspinesAllPaeonia suffruticosaRootPueraria thunbergianaRootPolygala tenuifoliaRootCoptis japonicaRhizomeAstragalus me
29、mbranaceusRootEucommia ulmoidesStem barkBupeuri falcatumRootAcanthopanax sessiliflorumBarkEpimedium koreaumAllMorus albaRoot barkArtium lappaFruitSpirodela polyrhizaAll第35页，共50页。Summary M.J.Noh,et.al.,op.cit For many materials,SFE yielded extracts with higher bioactivity than LSE.SFE was found to be
30、 more selective than LSE for target compounds.SFE conditions could be optimized to produce maximum levels of bioactivity.第36页，共50页。Drug ResiduesAnalyteMatrixReferenceSulfamethazineSwine Muscle TissueCross,et.al.Anabolic SteriodsBovine Tissue(Muscle and Liver)Houpalahti and HenionOpiatesHair,blood an
31、d tissueMultiple AuthorsTemazepamWhole BloodScott and OliverCocaine,benzoylecgonine,codeine and morphineHairBrewer,et.al.第37页，共50页。Study Summary Compared to a conventional SPE method,the SFE method was more efficient and gave cleaner extracts with recoveries above 80%K.S.Scott,J.S.Oliver,J.Anal.Toxi
32、col.21(1997)297.第38页，共50页。Supercritical Fluid Chromatography SFC is a separation technique similar to HPLC and GC where the mobile phase or carrier gas is replaced by a supercritical fluid第39页，共50页。Limitations of GC and HPLCGC Sample Limitations:Volatility Thermal stability Low molecular weight HPLC
33、 Analytical Limitations:No universal detector Low efficiency Low resolution 第40页，共50页。SFC Overcomes Limitations of GC and HPLC Extends molecular weight range of GC Lower operating temperature than GC Faster separation time than HPLC Higher separation efficiency than HPLC Universal detector can be us
34、ed,FID Both packed(HPLC-type)and GC-type columns can be used第41页，共50页。Carbon Dioxide,CO2has desirable properties as a SFC solvent Inexpensive Highly pure Very low UV absorbance NO FID background noise Low critical pressure and temperature Non-toxic Supercritical CO2 behaves as a nonpolar solvent suc
35、h as heptane Polar organic modifiers can be mixed with CO2 for more polar samples 第42页，共50页。SFC ApplicationsIndustrial:Synthetic oligomers,polymers/additives Surfactants(polyglycols)Oligo/polysaccharides,sucrose polyesters Pesticides Isocyanates Dyes Waxes第43页，共50页。SFC ApplicationsBiochemical:Steroi
36、ds Prostaglandins Fatty acids/lipids Antibiotics Drugs of abuse第44页，共50页。SFC ApplicationsFossil Fuels:Fractionation of petroleum and coal-derived fluids Hydrocarbon group analysis Simulated distillation第45页，共50页。Other Applications of Supercritical Fluid Supercritical Fluid Cleaning Supercritical Flu
37、id Drying Supercritical Fluid Reactions Micro Particles Formation Supercritical Water Oxidation System Others第46页，共50页。Applications of Supercritical Fluid Technologies in Taiwan IN THE PAST第47页，共50页。Applications of Supercritical Fluid Technologies in Taiwan AT PRESENT第48页，共50页。Applications of Supercritical Fluid Technologies in Taiwan IN THE FUTURE第49页，共50页。Thanks For Your Attention!林華經國立清華大學化工系學士國立清華大學生科所碩士國立清華大學化工系博士班友翔實業股份有限公司 儀器部經理引光生物科技有限公司 研發部經理第50页，共50页。
