同濟(jì)-有機(jī)物分析方法課件

溶液中有機(jī)物分析表征

溶液中有機(jī)物分析表征1主要內(nèi)容1.常用有機(jī)物表征方法2.有機(jī)物分離方法3.WR最新難降解有機(jī)物研究進(jìn)展4.沼液相關(guān)研究進(jìn)展主要內(nèi)容2常用有機(jī)物表征方法2.簡單、單一、未（已）知物1.復(fù)雜、混合、未（已）知物宏觀分析：COD,MW,FT-IR,NMR,UV,親疏水性等分離：GC,LC,SEC,IC,SPE分析方法FT-IR,MS,NMR等常用有機(jī)物表征方法2.簡單、單一、未（已）1.復(fù)雜、混合、未3MWdistributionanalysisMW

distributionanalysishasbeenextensivelyapplied

towastewatersamples,andissuitabletobeusedasafirststep

inanalyzingcomplexsamples.Sizedistributionanalysishasbeen

widelyused,normallycarriedoutusingultrafiltration(UF)or

sizeexclusionchromatography(SEC).UFmolecularweightcut-offs:1kDa,3kDa,5kDa,10kDa,100kDa,300kDaMWdistributionanalysisMWdis4SECisusefulforMWscreeningof

samplesbeforeusingUFtoobtaintheseparatedsamplefor

analysis.Thereare

alsosomedrawbacks:thechemicalinteraction

betweenthecolumnmaterials,thesolvent,andthe

organiccompoundsinSECcanoverestimate/underestimate

theMWprofile.Anotherdisadvantage

isthatthepeaksfromSECcanbeverydifficulttodifferentiate

betweeninsomesamples.SECisusefulforMWscreening5GPCiswhenthe

mobilephaseisanorganicsolvent,andGFCwhenthemobile

phaseisaqueous.gel-filtration-chromatography(GFC)gel-permeationchromatography(GPC)SECGPCiswhenthemobilephasei6UFismoresuitableforidentificationofcompounds,sincelargeamountsofthesamplecanbe

recoveredforfurthercharacterization.InUF,therearealsomany

factorssuchasmembraneporesizedistribution,sample

temperature,stirredcellpressure,pH,andmembranematerials

thatcaninfluencethetransportoforganicsthroughthe

membranes.UFismoresuitableforidenti7按照Alexetal.提出的有機(jī)碳物理尺寸概念，有機(jī)碳可分為顆粒有機(jī)碳(Particulateorganiccarbon,POC)、膠體有機(jī)碳(Colloidalorganiccarbon,COC)、微膠體有機(jī)碳(Finecolloidalorganiccarbon,FCOC)和溶解性有機(jī)碳(Dissolvedorganiccarbon,DOC)。POC能通過1.2μm濾膜但0.45μm濾膜截留；COC能通過0.45μm濾膜但被0.1μm濾膜截留；FCOC能通過0.1μm濾膜但被0.025μm濾膜截留；DOC能通過0.025μm濾膜。同濟(jì)-有機(jī)物分析方法課件8Evaluationofon-sitebiologicaltreatmentforlandfillleachates

anditsimpact:Asizedistributionstudy（WR）Evaluationofon-sitebiologic9Evaluationofon-sitebiologicaltreatmentforlandfillleachates

anditsimpact:Asizedistributionstudy（WR）Evaluationofon-sitebiologic10HPLC+SECcolumn:Thisseparationmethodwaseffectiveinrevealingthesimilaritiesanddifferencesbetweenmicrobialproductsfromtwodifferentactivatedsludgeplants,anddemonstratingthegenerationanddegradationofcompounds.Analyticalmethodsforsolublemicrobialproducts(SMP)andextracellularpolymers(ECP)inwastewatertreatmentsystems:Areview（WR）HPLC+SECcolumn:11HydrolysisofhighMWcompoundsForlowMWcompounds,GC-MScanbe

usedforidentificationbymatchingwithexistinglibraries.ForhighMWcompoundsitismorecomplicated.ThereisnoeasymethodtoidentifyhighMWcompoundsincomplex

tein/carbohydratesaminoacidsandmonosaccharideGC-MSHydrolysis:acidichydrolysis,microwaveradiationinducedhydrolysis,alkalinehydrolysis,andenzymatichydrolysishydrolysisHydrolysisofhighMWcompound12ProteinanalysisLowrymethodexcitation-emissionmatrixspectroscopy（EEM）resonancelights

cattering（共振光散射）proteomics（蛋白質(zhì)組）ion-exchangechromatographyProteinanalysisLowrymethod13Multi-dimensionalproteinidentificationtechnology(MudPIT)isbecomingaprevalentproteomicapproachduetoitshigh-throughput

separationsandaccuratemassdetection.In-solutiondigestioninvolvessimultaneousdigestionofhundredsorthousandsofproteins.（HPLC+tandemMS）Exploringhumanplasmaproteomestrategies:Highefficiency

in-solutiondigestionprotocolformulti-dimensionalprotein

identificationtechnology（JournalofChromatographyA）同濟(jì)-有機(jī)物分析方法課件14LowrymethodReaction:Cu2++peptidebonds（肽鍵）（pH＞7）Shortcoming:Proteinsareeasytodegrademonopeptides,polypeptides,and/orotherorganicpolymers,sotheidentificationof“proteins”isverylikelytobeanoverestimation.Theresultshouldbeconfirmedwithaproteomicstudy,orionexchangechromatography.Lowrymethod15ProteomicsProteomicsisthelarge-scalestudyofproteins,particularl

theirstructureandfunction.Steps：1.fractionationand/orproteinconcentration2.digestion3.separation(HPLC)4.detection(MS)However,thereisascarcityofproteomicstudiesappliedtoaerobicandanaerobicbiologicaltreatmentsystems.Proteomics16CarbohydrateanalysistitrationgravimetriccolorimetricenzymaticchromatographicCarbohydrateanalysistitration17ColorimetricuseAnthroneandPhenol-SulfuricAcidforquantificationofcarbohydratesChromatographicmethodsarealternativetechniquesfor

analyzingmonosaccharides(單糖)andoligosaccharides(多糖).Carbohydratescanbeseparatedbasedontheirdifferentialadsorptioncharacteristics.Thereareseveraltypesofcolumns

whichcanbeusedfortheanalysisofsugars,suchas

ion-exchange,amino,andreversed-phasecolumnsColorimetricuseAnthroneand18Usingaresonancelight-scatteringtechniquelowconcentrationofproteinsandcarbohydratescanbedetermined.CongoredandNeutralreddyeswereusedforthedeterminationofproteinsandcarbohydrates,respectively.Usingaresonancelight-scatte19UV-VisspectrometryHumicsubstancesin

bioreactoreffluentswerecharacterizedbyhavingahighdegree

ofaromaticity.specificUVA:TheratioofUVabsorbanceat254nmandtheDOCofthesample.Thisrepresentsameasureofaromaticityoftheorganicsinthesample.ResearchershavehaveusedspecificUVAtoevaluatethearomaticityofSMPsandECPsdandsuggestedthataromaticitycanincreaseafteranaerobictreatment.UV-VisspectrometryHumicsubst20Fluorescenceexcitation-emissionmatrix

spectroscopy(EEM)EEMhasbeenappliedasasubstituteforsomechemical

analysistoidentifycompoundsinthesamplebycomparing

theirEEMspectralfingerprints.EachEEMpresentsspectralinformationaboutthechemicalcompositionofthe

samples.EEMtechniquecanonlydetectfluorescence

activecompounds,whereasothercomponentssuchaspolysaccharides

werenotdetectableFluorescenceexcitation-emissi21Fouriertransforminfraredspectrometry(FTIR)FTIRhasbeenusedtodeterminethefunctional

groupsoforganicmatter,andtopredictthemajorcomponents

inwastewatertreatmentsystems.Butisnotsuitableforcompoundidentification.ItisnoteasytouseFTIRasaquantitativeanalyticaltechniqueduetothedifficultiesrelatedtosamplepreparation.Fouriertransforminfraredspe22NMRNMR可檢測的核素有很多,如1H、13C、15N、19F、23Na、31P等。由于大多數(shù)藥物都含有質(zhì)子,最常用的是1H-NMR和13C-NMR。對于一些含有氟、磷等原子的化合物,可借助于19F譜、31P譜為研究手段。

通過化學(xué)位移及其峰面積等可以得到相應(yīng)的官能團(tuán)、化學(xué)鍵信息。NMRNMR可檢測的核素有很多,如1H、13C、15N、192313C-NMR圖譜Characteristictransformationofhumicacidduring

photoelectrocatalysisprocessanditssubsequent

disinfectionbyproductformationpotential（WR）13C-NMR圖譜Characteristictransf24DOMfractionDOMfraction25Effectiveremovalofeffluentorganicmatter(EfOM)

frombio-treatedcokingwastewaterbyarecyclable

aminatedhyper-cross-linkedpolymer（WR）Effectiveremovalofeffluent26Contributionofeffluentorganicmatter(EfOM)to

ultrafiltration(UF)membranefouling:Isolation,

characterization,andfoulingeffectofEfOM

fractions（WR）Contributionofeffluentorgan27有機(jī)物提純、分離方法固相萃取毛細(xì)管電泳超臨界流體色譜有機(jī)物提純、分離方法固相萃取28固相萃取（SPE）

固相萃取是采用用固體物質(zhì)作為萃取劑,采用高效、高選擇性的固定相進(jìn)行萃取的樣品預(yù)處理技術(shù)。

SPE是一種吸附劑萃取，樣品通過填充吸附劑的一次性萃取柱，分析物和雜質(zhì)被保留在柱上，然后分別用選擇性溶劑去除雜質(zhì)，洗脫出分析物，從而達(dá)到分離的目的。SPE的分離模式主要取決于填充劑的類型和溶劑的性質(zhì)。固相萃?。⊿PE）固相萃取是采用用固體物質(zhì)作為萃取劑,29

SPE柱的填料粒徑（>40μm）要比HPLC填料（3~10μm）大。由于短的柱床和大的粒徑，SPE柱效比HPLC色譜柱低得多。因此，用SPE只能分開保留性質(zhì)有很大差別的化合物，且SPE柱是一次性使用。SPE柱的填料粒徑（>40μm）要比HPLC填料（330一個完整的固相萃取步驟包括固相萃取柱的預(yù)處理、上樣、洗去干擾物質(zhì)、洗脫及收集分析物四個步驟。一個完整的固相萃取步驟包括固相萃取柱的預(yù)處理、上樣、洗去干擾31毛細(xì)管電泳一、毛細(xì)管區(qū)帶電泳(CZE)capillaryzoneelectrophoresis二、膠束電動毛細(xì)管色譜(MEKC)micelleelectrokineticchromatography三、毛細(xì)管凝膠電泳（CGE）capillarygelelectrophoresis四、毛細(xì)管電色譜(CEC)capillaryelectrochromatography毛細(xì)管電泳一、毛細(xì)管區(qū)帶電泳(CZE)32毛細(xì)管區(qū)帶電泳

帶電粒子的遷移速度=電泳和電滲流速度的矢量和。正離子：兩種效應(yīng)的運(yùn)動方向一致，在負(fù)極最先流出；中性粒子：無電泳現(xiàn)象，受電滲流影響，在陽離子后流出；

陰離子：兩種效應(yīng)的運(yùn)動方向相反；ν電滲流>ν電泳時,陰離子在負(fù)極最后流出,在這種情況下,不但可以按類分離,同種類離子由于差速遷移被相互分離。這是最基本、應(yīng)用廣的分離模式；毛細(xì)管區(qū)帶電泳帶電粒子的遷移速度=電泳和電滲流速度的33

1.緩沖溶液中加入離子型表面活性劑，其濃度達(dá)到臨界濃度，形成一疏水內(nèi)核、外部帶電的膠束。

在電場力的作用下，膠束在柱中移動。膠束電動毛細(xì)管色譜1.緩沖溶液中加入離子型表面活性劑，其濃度達(dá)到臨界濃342.電泳流和電滲流的方向相反，且u電滲流>u電泳，負(fù)電膠束以較慢的速度向負(fù)極移動；3.中性分子在膠束相和溶液（水相）間分配，疏水性強(qiáng)的組分與膠束結(jié)合的較牢，流出時間長；4.可用來分離中性物質(zhì)，擴(kuò)展了高效毛細(xì)管電泳的應(yīng)用范圍；5.色譜與電泳分離模式的結(jié)合。2.電泳流和電滲流的方向相反，且u電滲流>u電泳，負(fù)35毛細(xì)管凝膠電泳

將聚丙烯酰胺等在毛細(xì)管柱內(nèi)交聯(lián)生成凝膠。其具有多孔性，類似分子篩的作用，試樣分子按大小分離。能夠有效減小組分?jǐn)U散，所得峰型尖銳，分離效率高。

蛋白質(zhì)、DNA等的電荷/質(zhì)量比與分子大小無關(guān)，CZE模式很難分離，采用CGE能獲得良好分離，DNA排序的重要手段。特點(diǎn)：抗對流性好，散熱性好，分離度極高。無膠篩分技術(shù)：采用低粘度的線性聚合物溶液代替高粘度交聯(lián)聚丙烯酰胺。柱便宜、易制備。毛細(xì)管凝膠電泳將聚丙烯酰胺等在毛細(xì)管柱內(nèi)交聯(lián)生成凝膠36毛細(xì)管電色譜將細(xì)粒徑固定相填充到毛細(xì)管中或在毛細(xì)管內(nèi)壁鍵合固定相，毛細(xì)管兩端施加高電壓，以電滲流驅(qū)動操作緩沖液。它也包含了電泳和色譜兩種機(jī)制，溶質(zhì)根據(jù)它們在流動相和固定相中的分配系數(shù)不同和自身淌度的差異得以分離。整合了毛細(xì)管電泳與微徑液相色譜的優(yōu)點(diǎn)，大大提高了樣品的分離能力，代表了分析學(xué)界高效微分離的趨勢，尤其適用于復(fù)雜生物及化學(xué)體系的研究。毛細(xì)管電色譜將細(xì)粒徑固定相填充到毛細(xì)管中或在毛細(xì)管內(nèi)壁鍵合固37超臨界流體色譜與高效液相色譜法比較實(shí)驗(yàn)證明SFC法的柱效一般比HPLC法要高：當(dāng)平均線速度為0.6cm·S-1時，SFC法的柱效可為HPLC法的3倍左右，在最小板高下載氣線速度是4倍左右；因此SFC法的分離時間也比HPLC法短。這是由于流體的低粘度使其流動速度比HPLC法快，有利于縮短分離時間。超臨界流體色譜與高效液相色譜法比較38

與氣相色譜法比較

出于流體的擴(kuò)散系數(shù)與粘度介于氣體和液體之間，因此SFC的譜帶展寬比GC要??；另外，SFC中流動相的作用類似LC中流動相，流體作流動相不僅載帶溶質(zhì)移動，而且與溶質(zhì)會產(chǎn)生相互作用力，參與選擇競爭。

如果我們把溶質(zhì)分子溶解在超臨界流體看作類似于揮發(fā)，這樣，大分子物質(zhì)的分壓很大，因此可應(yīng)用比GC低得多的溫度，實(shí)現(xiàn)對大分子物質(zhì)、熱不穩(wěn)定性化合物、高聚物等的有效分離。與氣相色譜法比較39

下圖描繪了SFC與其他色譜方法測定相對分子質(zhì)量范圍的比較。由圖看出SFC比起GC法測定相對分子質(zhì)量的范圍要大出好幾個數(shù)量級，基本與LC法相當(dāng)。下圖描繪了SFC與其他色譜方法測定相對分子質(zhì)量范圍的比較。40超臨界流體色譜的應(yīng)用

超臨界流體色譜法被廣泛應(yīng)用于糖類、蛋白質(zhì)、氨基酸、天然產(chǎn)物、藥物、表面活性劑、高聚物、多聚物、農(nóng)藥、炸藥和火箭推進(jìn)劑等物質(zhì)的分離和分析。超臨界流體色譜的應(yīng)用超臨界流體色譜法被廣泛應(yīng)用于糖類、蛋41WR中難降解有機(jī)物研究NumTitleContent1Removalofmicropollutantsduringtertiarywastewatertreatmentbybiofiltration:Roleofnitrifiersandremovalmechanisms反硝化細(xì)菌處理微污染2Treatmentofoliveoilmillwastewaterbycombinedprocesselectro-Fentonreactionandanaerobicdigestion電芬頓與厭氧消化處理油廠廢水3Textilewastewatertreatment:Aerobicgranularsludgevsactivatedsludgesystems好氧顆粒污泥與活性污泥處理印染廢水的對比4Biogenicmetalsfortheoxidativeandreductiveremovalofpharmaceuticals,biocidesandiodinatedcontrastmediainapolishingmembranebioreactor膜反應(yīng)器氧化還原處理制藥等廢水5Solarphotocatalyticoxidationofrecalcitrantnaturalmetabolicby-productsofamoxicillinbiodegradation先用微生物處理AMX，代謝產(chǎn)物沒有了抗菌性，但依然難降解，后采取光催化方式去除副產(chǎn)物。WR中難降解有機(jī)物研究NumTitleContent1Rem42NumTitleContent1Newchlorinatedamphetamine-type-stimulantsdisinfection-by-productsformedduringdrinkingwatertreatment自來水消毒中產(chǎn)生的新的副產(chǎn)物2Treatmentofmunicipalsolidwasteleachateusingasubmergedanaerobicmembranebioreactoratmesophilicandpsychrophilic

temperatures:AnalysisofrecalcitrantsinthepermeateusingGC-MS不同溫度對SAMBR處理污泥滲濾液的影響及其MS對難降解有機(jī)物的分析38.Photooxidationandsubsequentbiodegradabilityofrecalcitranttri-alkylphosphatesTCEPandTBPinwater光催化聯(lián)合生物方法處理難降解有機(jī)物4Characterisationandremovalofrecalcitrantsinreverseosmosisconcentratesfromwaterreclamationplants反滲透濃液中有機(jī)物特別是有色及含氮有機(jī)物的表征及去除，去除研究了鐵、鋁絮凝去除及其高級氧化去除。5Ultrasonictreatmentofwatercontaminatedwithibuprofen超聲處理含布洛芬的廢水NumTitleContent1Newchlorinate4344寫在最后成功的基礎(chǔ)在于好的學(xué)習(xí)習(xí)慣Thefoundationofsuccessliesingoodhabits44寫在最后成功的基礎(chǔ)在于好的學(xué)習(xí)習(xí)慣結(jié)束語當(dāng)你盡了自己的最大努力時，失敗也是偉大的，所以不要放棄，堅持就是正確的。WhenYouDoYourBest,FailureIsGreat,SoDon'TGiveUp,StickToTheEnd演講人：XXXXXX

時間：XX年XX月XX日

結(jié)束語4546寫在最后成功的基礎(chǔ)在于好的學(xué)習(xí)習(xí)慣Thefoundationofsuccessliesingoodhabits46寫在最后成功的基礎(chǔ)在于好的學(xué)習(xí)習(xí)慣結(jié)束語當(dāng)你盡了自己的最大努力時，失敗也是偉大的，所以不要放棄，堅持就是正確的。WhenYouDoYourBest,FailureIsGreat,SoDon'TGiveUp,StickToTheEnd演講人：XXXXXX

時間：XX年XX月XX日

結(jié)束語47溶液中有機(jī)物分析表征

溶液中有機(jī)物分析表征48主要內(nèi)容1.常用有機(jī)物表征方法2.有機(jī)物分離方法3.WR最新難降解有機(jī)物研究進(jìn)展4.沼液相關(guān)研究進(jìn)展主要內(nèi)容49常用有機(jī)物表征方法2.簡單、單一、未（已）知物1.復(fù)雜、混合、未（已）知物宏觀分析：COD,MW,FT-IR,NMR,UV,親疏水性等分離：GC,LC,SEC,IC,SPE分析方法FT-IR,MS,NMR等常用有機(jī)物表征方法2.簡單、單一、未（已）1.復(fù)雜、混合、未50MWdistributionanalysisMW

distributionanalysishasbeenextensivelyapplied

towastewatersamples,andissuitabletobeusedasafirststep

inanalyzingcomplexsamples.Sizedistributionanalysishasbeen

widelyused,normallycarriedoutusingultrafiltration(UF)or

sizeexclusionchromatography(SEC).UFmolecularweightcut-offs:1kDa,3kDa,5kDa,10kDa,100kDa,300kDaMWdistributionanalysisMWdis51SECisusefulforMWscreeningof

samplesbeforeusingUFtoobtaintheseparatedsamplefor

analysis.Thereare

alsosomedrawbacks:thechemicalinteraction

betweenthecolumnmaterials,thesolvent,andthe

organiccompoundsinSECcanoverestimate/underestimate

theMWprofile.Anotherdisadvantage

isthatthepeaksfromSECcanbeverydifficulttodifferentiate

betweeninsomesamples.SECisusefulforMWscreening52GPCiswhenthe

mobilephaseisanorganicsolvent,andGFCwhenthemobile

phaseisaqueous.gel-filtration-chromatography(GFC)gel-permeationchromatography(GPC)SECGPCiswhenthemobilephasei53UFismoresuitableforidentificationofcompounds,sincelargeamountsofthesamplecanbe

recoveredforfurthercharacterization.InUF,therearealsomany

factorssuchasmembraneporesizedistribution,sample

temperature,stirredcellpressure,pH,andmembranematerials

thatcaninfluencethetransportoforganicsthroughthe

membranes.UFismoresuitableforidenti54按照Alexetal.提出的有機(jī)碳物理尺寸概念，有機(jī)碳可分為顆粒有機(jī)碳(Particulateorganiccarbon,POC)、膠體有機(jī)碳(Colloidalorganiccarbon,COC)、微膠體有機(jī)碳(Finecolloidalorganiccarbon,FCOC)和溶解性有機(jī)碳(Dissolvedorganiccarbon,DOC)。POC能通過1.2μm濾膜但0.45μm濾膜截留；COC能通過0.45μm濾膜但被0.1μm濾膜截留；FCOC能通過0.1μm濾膜但被0.025μm濾膜截留；DOC能通過0.025μm濾膜。同濟(jì)-有機(jī)物分析方法課件55Evaluationofon-sitebiologicaltreatmentforlandfillleachates

anditsimpact:Asizedistributionstudy（WR）Evaluationofon-sitebiologic56Evaluationofon-sitebiologicaltreatmentforlandfillleachates

anditsimpact:Asizedistributionstudy（WR）Evaluationofon-sitebiologic57HPLC+SECcolumn:Thisseparationmethodwaseffectiveinrevealingthesimilaritiesanddifferencesbetweenmicrobialproductsfromtwodifferentactivatedsludgeplants,anddemonstratingthegenerationanddegradationofcompounds.Analyticalmethodsforsolublemicrobialproducts(SMP)andextracellularpolymers(ECP)inwastewatertreatmentsystems:Areview（WR）HPLC+SECcolumn:58HydrolysisofhighMWcompoundsForlowMWcompounds,GC-MScanbe

usedforidentificationbymatchingwithexistinglibraries.ForhighMWcompoundsitismorecomplicated.ThereisnoeasymethodtoidentifyhighMWcompoundsincomplex

tein/carbohydratesaminoacidsandmonosaccharideGC-MSHydrolysis:acidichydrolysis,microwaveradiationinducedhydrolysis,alkalinehydrolysis,andenzymatichydrolysishydrolysisHydrolysisofhighMWcompound59ProteinanalysisLowrymethodexcitation-emissionmatrixspectroscopy（EEM）resonancelights

cattering（共振光散射）proteomics（蛋白質(zhì)組）ion-exchangechromatographyProteinanalysisLowrymethod60Multi-dimensionalproteinidentificationtechnology(MudPIT)isbecomingaprevalentproteomicapproachduetoitshigh-throughput

separationsandaccuratemassdetection.In-solutiondigestioninvolvessimultaneousdigestionofhundredsorthousandsofproteins.（HPLC+tandemMS）Exploringhumanplasmaproteomestrategies:Highefficiency

in-solutiondigestionprotocolformulti-dimensionalprotein

identificationtechnology（JournalofChromatographyA）同濟(jì)-有機(jī)物分析方法課件61LowrymethodReaction:Cu2++peptidebonds（肽鍵）（pH＞7）Shortcoming:Proteinsareeasytodegrademonopeptides,polypeptides,and/orotherorganicpolymers,sotheidentificationof“proteins”isverylikelytobeanoverestimation.Theresultshouldbeconfirmedwithaproteomicstudy,orionexchangechromatography.Lowrymethod62ProteomicsProteomicsisthelarge-scalestudyofproteins,particularl

theirstructureandfunction.Steps：1.fractionationand/orproteinconcentration2.digestion3.separation(HPLC)4.detection(MS)However,thereisascarcityofproteomicstudiesappliedtoaerobicandanaerobicbiologicaltreatmentsystems.Proteomics63CarbohydrateanalysistitrationgravimetriccolorimetricenzymaticchromatographicCarbohydrateanalysistitration64ColorimetricuseAnthroneandPhenol-SulfuricAcidforquantificationofcarbohydratesChromatographicmethodsarealternativetechniquesfor

analyzingmonosaccharides(單糖)andoligosaccharides(多糖).Carbohydratescanbeseparatedbasedontheirdifferentialadsorptioncharacteristics.Thereareseveraltypesofcolumns

whichcanbeusedfortheanalysisofsugars,suchas

ion-exchange,amino,andreversed-phasecolumnsColorimetricuseAnthroneand65Usingaresonancelight-scatteringtechniquelowconcentrationofproteinsandcarbohydratescanbedetermined.CongoredandNeutralreddyeswereusedforthedeterminationofproteinsandcarbohydrates,respectively.Usingaresonancelight-scatte66UV-VisspectrometryHumicsubstancesin

bioreactoreffluentswerecharacterizedbyhavingahighdegree

ofaromaticity.specificUVA:TheratioofUVabsorbanceat254nmandtheDOCofthesample.Thisrepresentsameasureofaromaticityoftheorganicsinthesample.ResearchershavehaveusedspecificUVAtoevaluatethearomaticityofSMPsandECPsdandsuggestedthataromaticitycanincreaseafteranaerobictreatment.UV-VisspectrometryHumicsubst67Fluorescenceexcitation-emissionmatrix

spectroscopy(EEM)EEMhasbeenappliedasasubstituteforsomechemical

analysistoidentifycompoundsinthesamplebycomparing

theirEEMspectralfingerprints.EachEEMpresentsspectralinformationaboutthechemicalcompositionofthe

samples.EEMtechniquecanonlydetectfluorescence

activecompounds,whereasothercomponentssuchaspolysaccharides

werenotdetectableFluorescenceexcitation-emissi68Fouriertransforminfraredspectrometry(FTIR)FTIRhasbeenusedtodeterminethefunctional

groupsoforganicmatter,andtopredictthemajorcomponents

inwastewatertreatmentsystems.Butisnotsuitableforcompoundidentification.ItisnoteasytouseFTIRasaquantitativeanalyticaltechniqueduetothedifficultiesrelatedtosamplepreparation.Fouriertransforminfraredspe69NMRNMR可檢測的核素有很多,如1H、13C、15N、19F、23Na、31P等。由于大多數(shù)藥物都含有質(zhì)子,最常用的是1H-NMR和13C-NMR。對于一些含有氟、磷等原子的化合物,可借助于19F譜、31P譜為研究手段。

通過化學(xué)位移及其峰面積等可以得到相應(yīng)的官能團(tuán)、化學(xué)鍵信息。NMRNMR可檢測的核素有很多,如1H、13C、15N、197013C-NMR圖譜Characteristictransformationofhumicacidduring

photoelectrocatalysisprocessanditssubsequent

disinfectionbyproductformationpotential（WR）13C-NMR圖譜Characteristictransf71DOMfractionDOMfraction72Effectiveremovalofeffluentorganicmatter(EfOM)

frombio-treatedcokingwastewaterbyarecyclable

aminatedhyper-cross-linkedpolymer（WR）Effectiveremovalofeffluent73Contributionofeffluentorganicmatter(EfOM)to

ultrafiltration(UF)membranefouling:Isolation,

characterization,andfoulingeffectofEfOM

fractions（WR）Contributionofeffluentorgan74有機(jī)物提純、分離方法固相萃取毛細(xì)管電泳超臨界流體色譜有機(jī)物提純、分離方法固相萃取75固相萃?。⊿PE）

固相萃取是采用用固體物質(zhì)作為萃取劑,采用高效、高選擇性的固定相進(jìn)行萃取的樣品預(yù)處理技術(shù)。

SPE是一種吸附劑萃取，樣品通過填充吸附劑的一次性萃取柱，分析物和雜質(zhì)被保留在柱上，然后分別用選擇性溶劑去除雜質(zhì)，洗脫出分析物，從而達(dá)到分離的目的。SPE的分離模式主要取決于填充劑的類型和溶劑的性質(zhì)。固相萃?。⊿PE）固相萃取是采用用固體物質(zhì)作為萃取劑,76

SPE柱的填料粒徑（>40μm）要比HPLC填料（3~10μm）大。由于短的柱床和大的粒徑，SPE柱效比HPLC色譜柱低得多。因此，用SPE只能分開保留性質(zhì)有很大差別的化合物，且SPE柱是一次性使用。SPE柱的填料粒徑（>40μm）要比HPLC填料（377一個完整的固相萃取步驟包括固相萃取柱的預(yù)處理、上樣、洗去干擾物質(zhì)、洗脫及收集分析物四個步驟。一個完整的固相萃取步驟包括固相萃取柱的預(yù)處理、上樣、洗去干擾78毛細(xì)管電泳一、毛細(xì)管區(qū)帶電泳(CZE)capillaryzoneelectrophoresis二、膠束電動毛細(xì)管色譜(MEKC)micelleelectrokineticchromatography三、毛細(xì)管凝膠電泳（CGE）capillarygelelectrophoresis四、毛細(xì)管電色譜(CEC)capillaryelectrochromatography毛細(xì)管電泳一、毛細(xì)管區(qū)帶電泳(CZE)79毛細(xì)管區(qū)帶電泳

帶電粒子的遷移速度=電泳和電滲流速度的矢量和。正離子：兩種效應(yīng)的運(yùn)動方向一致，在負(fù)極最先流出；中性粒子：無電泳現(xiàn)象，受電滲流影響，在陽離子后流出；

陰離子：兩種效應(yīng)的運(yùn)動方向相反；ν電滲流>ν電泳時,陰離子在負(fù)極最后流出,在這種情況下,不但可以按類分離,同種類離子由于差速遷移被相互分離。這是最基本、應(yīng)用廣的分離模式；毛細(xì)管區(qū)帶電泳帶電粒子的遷移速度=電泳和電滲流速度的80

1.緩沖溶液中加入離子型表面活性劑，其濃度達(dá)到臨界濃度，形成一疏水內(nèi)核、外部帶電的膠束。

在電場力的作用下，膠束在柱中移動。膠束電動毛細(xì)管色譜1.緩沖溶液中加入離子型表面活性劑，其濃度達(dá)到臨界濃812.電泳流和電滲流的方向相反，且u電滲流>u電泳，負(fù)電膠束以較慢的速度向負(fù)極移動；3.中性分子在膠束相和溶液（水相）間分配，疏水性強(qiáng)的組分與膠束結(jié)合的較牢，流出時間長；4.可用來分離中性物質(zhì)，擴(kuò)展了高效毛細(xì)管電泳的應(yīng)用范圍；5.色譜與電泳分離模式的結(jié)合。2.電泳流和電滲流的方向相反，且u電滲流>u電泳，負(fù)82毛細(xì)管凝膠電泳

將聚丙烯酰胺等在毛細(xì)管柱內(nèi)交聯(lián)生成凝膠。其具有多孔性，類似分子篩的作用，試樣分子按大小分離。能夠有效減小組分?jǐn)U散，所得峰型尖銳，分離效率高。

蛋白質(zhì)、DNA等的電荷/質(zhì)量比與分子大小無關(guān)，CZE模式很難分離，采用CGE能獲得良好分離，DNA排序的重要手段。特點(diǎn)：抗對流性好，散熱性好，分離度極高。無膠篩分技術(shù)：采用低粘度的線性聚合物溶液代替高粘度交聯(lián)聚丙烯酰胺。柱便宜、易制備。毛細(xì)管凝膠電泳將聚丙烯酰胺等在毛細(xì)管柱內(nèi)交聯(lián)生成凝膠83毛細(xì)管電色譜將細(xì)粒徑固定相填充到毛細(xì)管中或在毛細(xì)管內(nèi)壁鍵合固定相，毛細(xì)管兩端施加高電壓，以電滲流驅(qū)動操作緩沖液。它也包含了電泳和色譜兩種機(jī)制，溶質(zhì)根據(jù)它們在流動相和固定相中的分配系數(shù)不同和自身淌度的差異得以分離。整合了毛細(xì)管電泳與微徑液相色譜的優(yōu)點(diǎn)，大大提高了樣品的分離能力，代表了分析學(xué)界高效微分離的趨勢，尤其適用于復(fù)雜生物及化學(xué)體系的研究。毛細(xì)管電色譜將細(xì)粒徑固定相填充到毛細(xì)管中或在毛細(xì)管內(nèi)壁鍵合固84超臨界流體色譜與高效液相色譜法比較實(shí)驗(yàn)證明SFC法的柱效一般比HPLC法要高：當(dāng)平均線速度為0.6cm·S-1時，SFC法的柱效可為HPLC法的3倍左右，在最小板高下載氣線速度是4倍左右；因此SFC法的分離時間也比HPLC法短。這是由于流體的低粘度使其流動速度比HPLC法快，有利于縮短分離時間。超臨界流體色譜與高效液相色譜法比較85

與氣相色譜法比較

出于流體的擴(kuò)散系數(shù)與粘度介于氣體和液體之間，因此SFC的譜帶展寬比GC要??；另外，SFC中流動相的作用類似LC中流動相，流體作流動相不僅載帶溶質(zhì)移動，而且與溶質(zhì)會產(chǎn)生相互作用力，參與