液相微萃取在藥物分析中的應(yīng)用

液相微萃取技術(shù)在藥物分析中的應(yīng)用樣品的前處理是分析化學的一個重要環(huán)節(jié)，甚至是制約復雜樣品分析的關(guān)鍵環(huán)節(jié)。因此尋找一種高效、快速、簡便、環(huán)境友好的前處理方法，成為復雜樣品分析必需解決的問題，一直是分析化學的研究熱點。液-液萃取作為經(jīng)典的萃取方法，在樣品分離上起著重要的作用，但其萃取耗時長，操作步驟多，消耗大量有機溶劑，易造成環(huán)境的二次污染[張紅，陳玲，陳皓，彭中良．液相微萃取技術(shù)及其在環(huán)境水樣預(yù)處理中的應(yīng)用[J]．環(huán)境監(jiān)測管理與技術(shù)，2004張紅，陳玲，陳皓，彭中良．液相微萃取技術(shù)及其在環(huán)境水樣預(yù)處理中的應(yīng)用[J]．環(huán)境監(jiān)測管理與技術(shù)，2004，16(6)：8-11．ProsenH，L．Zupancic-Kralj．Solid·phaseextractionTrends．AnaLChem，1999，18：272-282．20世紀90年代中后期，He和Lee[HeY,LeeHK.AnalChem,1997,69:4634]、Jeannot和Cantwell[JeannotMA,CantweellFF.AnalChem,1997,69:235]，分別提出了較為成熟的液相微萃取(LiquidPhaseMicroextraction，LPME)方法。其基本原理是目標分析物在樣品與微升級的萃取溶劑之間達到分配平衡,從而實現(xiàn)溶質(zhì)的萃取和凈化。即將樣品前處理所涉及的多個步驟（萃取、濃縮、凈化）以及直接進樣進行GC分析加以組合，大大簡化了樣品前處理的操作，同時也實現(xiàn)了待測組分的富集。液相微萃取方法富集倍數(shù)可高達1000倍以上，操作簡便、萃取速度快、操作成本低、不污環(huán)境，便于與GC、HPLC及CE等高效分離檢測手段聯(lián)用。通過對LPME技術(shù)的不斷發(fā)展與改進，這種新型的萃取方法已成為現(xiàn)代儀器分析領(lǐng)域一種非常重要的樣品前處理技術(shù)，在環(huán)境分析[Lopez-DariasJ,German-HernandezM,PinoV,AfonsoAM.Talanta,2010,80:1611,ZhangJ,LeeHK.Talanta,2010,81:537,HangY,WuC.AnalChimActa,2010,661:161]、食品分析[HeY,LeeHK.AnalChem,1997,69:4634JeannotMA,CantweellFF.AnalChem,1997,69:235Lopez-DariasJ,German-HernandezM,PinoV,AfonsoAM.Talanta,2010,80:1611ZhangJ,LeeHK.Talanta,2010,81:537HangY,WuC.AnalChimActa,2010,661:161LingD,XieH,HeY,etal.JChromatogrA,2010,1217:1856.HashemiM,HabibiA,JahanshahiN.FoodChem.2011,124:1258AmvraziEG,Papade-PsyllouAT,TsiropoulosNG..IntJEnvironAnalChem,2010,90:245.液相微萃取的基本原理1.1.相平衡理論液相微萃取的基本原理與經(jīng)典的液-液萃取相似，可以通過相平衡理論解釋。當目標分析物A在溶劑水（w）和萃取溶劑（o）之間進行萃取，達到萃取平衡時，目標分析物A在兩相中的分配系數(shù)K為一常數(shù)，可表示為：(1)式中Co,為萃取平衡時目標物A在有機相中的濃度,Cw,目標物A在水相中的濃度。根據(jù)質(zhì)量平衡關(guān)系式,則：（2）式中為Cw,initial為樣本溶液中目標物A的最初濃度,Vw為樣本溶液的體積,Vo為有機相體積。由式（1）、式（2）可得：（3）1.2.富集倍數(shù)（EF）富集倍數(shù)（EF）是指當達到萃取平衡時，目標分析物A在萃取溶劑的濃度（Co,eq）和樣品相中的最初濃度（Cw,initial）之比。即：（4）將式（2）、式（3）代入得：（5）通過公式(5)可知,要獲得較高的富集倍數(shù)，可通過降低有機相與水相的體積比Vo/Vw，或選擇具有較高分配系數(shù)的兩相組合來實現(xiàn)。因此，液相微萃取適合將非極性或中等極性的物質(zhì)從水相中富集到有機萃取溶劑中。1.3.萃取回收率（ER）萃取回收率（ER）為當達到萃取平衡時，目標分析物A被萃取到有機相得量（no）占樣品相中分析物初始量（nw,initial）之比。即：（6）將式（1）代入得：（7）在液相微萃取中，Vo較小，Vw較大，分析物的Kow適中，致使KowVo＜＜Vw，此時上式可表達為：。由此可見在降低有機相與水相的體積比Vo/VW，提高富集倍數(shù)的同時也會降低萃取效率。因此需要通過試驗調(diào)整有機相與水相的體積比Vo/VW，達到合適的富集倍數(shù)和萃取回收率。液相微萃取的形式經(jīng)過近十幾年研究，液相微萃取技術(shù)發(fā)展出多種不同的萃取方式。根據(jù)萃取過程的狀態(tài)可以分為靜態(tài)液相微萃取(Staticliquidphasemicroextraction，S-LPME)[L．Zhao，H．K．Lee．Applicationofstaticliquid．phasemicroextractionoftheanalysisoforganochlorinepesticidesinwater．ZChromatogrA，2001，919：381-388．]、動態(tài)液相微萃取(Dynamicliquidphasemicroextraction，D-LPME)[L-Hou,H.KLce.Applicationofstaticanddynamicliquid-phasemicroextractioninthedeterminationofpolycyclicaromatichydrocarbos.J.Chromatogr．A，2002，976(1-2)：377-385.]、連續(xù)流動液相微萃取(Continuousflowmicrocxtraction，CFME)[W.Liu,H.K.Lee.Continuous-flowmicroextractionexceeding1000foldconcentrationofdiluteanalytes．AnalChem．，2000,72(18)：4462-4467.]；根據(jù)萃取劑與樣品的作用形式可以分為直接浸入式液相微萃取(Directimmersionliquidphasemieroextraction，DI-LPME)E.M.Giotl,D.C.Skalkos,Y.C.Fiamegos,C.D.Stalikas.Single-dropliquid-phasemicroextractionforthedeterminationofhyperlein,pseudohyperieinandhyperforininbiologicalfluidsbyhighperformanceliquidchromatography.L．Zhao，H．K．Lee．Applicationofstaticliquid．phasemicroextractionoftheanalysisoforganochlorinepesticidesinwater．ZChromatogrA，2001，919：381-388．L-Hou,H.KLce.Applicationofstaticanddynamicliquid-phasemicroextractioninthedeterminationofpolycyclicaromatichydrocarbos.J.Chromatogr．A，2002，976(1-2)：377-385.W.Liu,H.K.Lee.Continuous-flowmicroextractionexceeding1000foldconcentrationofdiluteanalytes．AnalChem．，2000,72(18)：4462-4467.E.M.Giotl,D.C.Skalkos,Y.C.Fiamegos,C.D.Stalikas.Single-dropliquid-phasemicroextractionforthedeterminationofhyperlein,pseudohyperieinandhyperforininbiologicalfluidsbyhighperformanceliquidchromatography.J.Chromatogr.A,2005,1093(1-2):l-10.LorenaVidal,AntonioCanals,NicolasKalogerakis,ElefleriaPsillakis.Headspacesingle-dropmicroextractionfortheanalysisofchlorobenzenesinwatersamples.J.Chromatogr.A,2005,1089(112):25-30.H.Liu,EK.Dasgupta.Analyticalchemistryinadrop.Solventextracioninamicrodrop.AnalChem.,1996,68(11):1817-1821.S.Pedersen-Bjegaard,K.E.Rasmussen.Liquid-Liquid-Liquidmicroextrationforpreparationofbiologicalfluidspriortocapillaryelectrophoresis.AnalChem.,1999,71(14):2650-2656.單滴液相微萃取（Single-Dropliquidphasemieroextraction，SDLPME）單滴液相微萃取是最早開發(fā)的液相微萃取方法。其主要方式見圖1，是將1滴與水不相溶的有機溶劑液滴浸沒在流動著的大水滴或浸入到被攪動著的樣品溶液中對分析物進行萃取。通常使用微量注射器作為有機溶劑的載體，并在萃取完成后吸取萃取溶劑用于進樣。在此基礎(chǔ)之上發(fā)展得而來的動態(tài)單滴液相微萃取則是將有機溶劑抽進微量注射器中隨后將含被測物的水溶液（樣品溶液）也吸入裝有有機溶劑的注射器中，反復推拉注射器活塞進行萃取。在反復推拉萃取的同時有機溶劑在微量注射器內(nèi)壁形成一層有機溶劑薄膜，在每一次推拉萃取過程中，注射器腔內(nèi)的萃取溶劑和腔壁有機液膜不斷地進行更新和交換，實現(xiàn)了動態(tài)單滴液相微萃取。動態(tài)單滴液相微萃取不僅增加了被測物在兩相中的擴散速度，并且提高了其富集倍數(shù)。KamleshShrivas等利用單滴液相微萃取分析生物樣本中的奎寧時，測得動態(tài)液相微萃取的富集倍數(shù)及靜態(tài)的富集倍數(shù)分別為14和3，最低檢出限分別為0.15和0.8μM[KamleshShrivas,Hui-FenWu.Quantitativebioanalysisofquininebyatmosphericpressure-matrixassistedlaserdesorption/ionizationmassspeKamleshShrivas,Hui-FenWu.Quantitativebioanalysisofquininebyatmosphericpressure-matrixassistedlaserdesorption/ionizationmassspectrometrycombinedwithdynamicdrop-to-dropsolventmicroextraction.AnalyticaChimicaActa2007,605:153-158.圖SEQ圖\*ARABIC1單滴液相微萃取示意圖單滴液相微萃取適合用于萃取分子量相對較大、熔點和沸點較高、在水中有適當溶解度的化合物，這類化合物辛醇-水分配系數(shù)較大（KOW約在2000附近）。常用沸點相對較低的有機溶劑萃?。ㄈ纾杭妆健⒍妆?、正己烷），這類溶劑萃取后可直接進入GC，進行分析。而大多數(shù)極性分子（如酮類、醇類和胺類）極易與溶劑形成氫鍵或產(chǎn)生偶極作用，能被醇類溶劑有效萃取[KokosaJM,PrzyjaznyA,JeannotMA.SolventMicroextraction:TheoryandPractice.NewJersey:JohnWiley&Sons,2009:101-155.]。萃取得到的分析KokosaJM,PrzyjaznyA,JeannotMA.SolventMicroextraction:TheoryandPractice.NewJersey:JohnWiley&Sons,2009:101-155.單滴液相微萃取具有溶劑用量少、儀器設(shè)備簡單、操作快速等優(yōu)點。但是該方法適合于萃取較為潔凈的液體樣品，對樣品基質(zhì)中含有固體顆粒、鹽、不溶性的有機物或能乳化有機溶劑的可溶性蛋白等的復雜基質(zhì)樣品的萃取效果較差。而且懸在微量注射器針頭的有機液滴在樣品攪拌時易于脫落。因此，單滴液相微萃取在實際工作中的應(yīng)用受到了限制。在用于體內(nèi)藥物分析時，最適合用于尿液的分析。對于以離子形式存在的被測物，Ma和Cantwell[M.Ma,F.F.Cantwell,Anal.Chem.1999,71:388–M.Ma,F.F.Cantwell,Anal.Chem.1999,71:388–393.圖SEQ圖\*ARABIC2液-液-液微萃取示意圖表1單滴液相微萃取在藥物分析中的應(yīng)用分析樣品被測物萃取模式分析方法萃取溶劑參考文獻尿樣、血液奎寧動態(tài)SDLPMEAP-MALDI/MS間二甲苯[NOTEREF_Ref391198347\h18]尿樣異丙嗪去甲替林丙咪嗪異丁嗪異丙叉丙酮SDLPMECE甲苯[Hui-FenWua,SureshKumarKailasa,Ja-YiYanHui-FenWua,SureshKumarKailasa,Ja-YiYan,etal.Comparisonofsingle-dropmicroextractionwithmicrovolumepipetteextractiondirectlycoupledwithcapillaryelectrophoresisforextractionandseparationoftricyclicantidepressantdrugs.J.IndustrialandEngineeringChemistry2014,20:2071–2076.尿樣咪唑斯汀馬來酸氯苯那敏非尼拉敏三相SDLPMEHPLC-UV接收相：0.1mol/L磷酸溶液中介相：正辛醇[WenhuaGao,YunshengChen,GaopanChen,etal.,Traceanalysisofthreeantihistaminesinhumanurinebyon-linesingledropliquid–liquid–liquidmicroextractioncoupledtosweepingmicellarelectrokineticchromatographyanditsapplicationtopharmacokineticstudy,[J]WenhuaGao,YunshengChen,GaopanChen,etal.,Traceanalysisofthreeantihistaminesinhumanurinebyon-linesingledropliquid–liquid–liquidmicroextractioncoupledtosweepingmicellarelectrokineticchromatographyanditsapplicationtopharmacokineticstudy,[J].JournalofChromatographyB,904(2012)121-127血漿阿托伐他丁三相SDLPMEHPLC-UV接收相：pH=11的磷酸鹽緩沖液中介相：正辛醇[HadiFarahani,ParvizNorouzi,AbolghasemBeheshti,etal.HadiFarahani,ParvizNorouzi,AbolghasemBeheshti,etal.,Quantitationofatorvastatininhumanplasmausingdirectlysuspendedacceptordropletinliquid–liquid–liquidmicroextractionandhigh-performanceliquidchromatography-ultravioletdetection,[J].Talanta,8(2009)1001-1006,尿液煙酸SDLPMEMALDI–TOF甲苯[KamleshShrivas,DeveshKumarPatel,Quantitativedeterminationofnicotinicacidinmicrolitervolumeofurinesamplebydrop-to-dropsolventmicroextractioncoupledtomatrixassistedlaserdesorption/ionizationmassspectrometry,[J].KamleshShrivas,DeveshKumarPatel,Quantitativedeterminationofnicotinicacidinmicrolitervolumeofurinesamplebydrop-to-dropsolventmicroextractioncoupledtomatrixassistedlaserdesorption/ionizationmassspectrometry,[J].SpectrochimicaActaPartA:MolecularandBiomolecularSpectroscopy,78(2011)253-257中空纖維液相微萃?。╤ollow-fiberliquidphasemieroextraction，HF-LPME）為了克服單滴液相微萃取的缺點（萃取液滴易于脫落，不適合復雜基質(zhì)的分析），Pedersen等人[S.Pedersen-Bjergaard,K.E.Rasmussen,Anal.Chem.71(1999)2650]提出了以多孔纖維膜為基礎(chǔ)的新型液相微萃取，裝置見圖3。該方法將數(shù)微升的萃取溶劑（接受相）注入到浸沒于水溶液（樣品相）中的中空纖維管腔內(nèi)進行萃取，達到萃取平衡后用微量注射器吸管腔內(nèi)的接受相用于分析測定。這樣較好地保護了萃取溶劑，同時由于中空纖維的多孔性,增加了溶劑與樣品接觸的表面積,從而提高了萃取效率,減小了溶劑損失,降低了復雜基質(zhì)的影響,實現(xiàn)了在萃取的同時凈化樣品。S.Pedersen-Bjergaard,K.E.Rasmussen,Anal.Chem.71(1999)2650圖SEQ圖\*ARABIC3中空纖維液相微萃取示意圖（ａ）（ｂ）圖４兩相（ａ）和三相（ｂ）中空纖維液相微萃取原理圖兩相中空纖維液相微萃取圖4.a所示為兩相中空纖維液相微萃取形式，中空纖維管腔內(nèi)部的接受相和吸附在中空纖維壁孔中的液膜為同一種有機溶劑，水相中的被測物質(zhì)由于萃取融入壁孔中及管腔內(nèi)的有機溶劑中，達到富集、精華的作用。溶劑的體積通為10-20μl，萃取之后，吸取接受相直接用于GC分析；也有少數(shù)情況為吸取接受相之后揮干溶劑，在用合適的溶劑溶劑用于HPLC或CE分析。兩相中空纖維液相微萃取適合于多數(shù)在有機相中溶解度大于其在水相中溶解度的弱極性物質(zhì)。兩相中空纖維液相微萃取的萃取溶劑應(yīng)滿足三個條件[YadollahYamini,CurtT.Reimann,AlirezaVatanara,etal.,ExtractionandPreconYadollahYamini,CurtT.Reimann,AlirezaVatanara,etal.,ExtractionandPreconcentrationofSalbutamolandTerbutalinefromAqueousSamplesUsingHollowFiberSupportedLiquidMembraneContainingAnionicCarrier[J].JournalofChromatographyA,2006,1124(1):57QinXiao等[QinXiao,BinHu.Hollowfiber-liquidphasemicroextractioncombinedwithgaschromatographyforthedeterminationofphenothiazinedrugsinurine.J.ChromatographyB.2010,878:1599-1640.]建立了兩相液相微萃取方法用于分析尿液中的四種酚噻嗪類藥物。在他們的試驗中pH=9.0的3mL樣品溶液被置于玻璃小瓶中，以甲苯為萃取溶劑，將在甲苯中浸泡過的15mm中空纖維浸沒在樣品溶液進行萃取。萃取溫度為40℃，萃取時間為10分鐘，磁力攪拌器速度為1000rpm。富集倍數(shù)為98~141。QinXiao,BinHu.Hollowfiber-liquidphasemicroextractioncombinedwithgaschromatographyforthedeterminationofphenothiazinedrugsinurine.J.ChromatographyB.2010,878:1599-1640.三相中空纖維液相微萃取圖4.b所示為三相中空纖維液相微萃取形式，纖維壁上的小孔中充滿了有機相作為液膜，而纖維內(nèi)部通常為水溶液接受相。在這種體系中分析物先從樣品相被萃取到有機溶劑液膜相,再被置于纖維內(nèi)部的接受相進行反萃取。該方法適合用于酸性或者堿性可解離物質(zhì)的前處理。通過調(diào)節(jié)供相和接受相的pH值，實現(xiàn)被測物質(zhì)從供相到接受相的轉(zhuǎn)移。對在于堿性物質(zhì)，供相的pH值應(yīng)調(diào)節(jié)至堿性，被測物處理游離狀態(tài)，抑制其在水相中的溶解。而接受相的pH值相對較低，增加被測物質(zhì)其中的溶解度。從而促使堿性物質(zhì)從供相向接受相轉(zhuǎn)移。Ugland等[HegeGrefslieUgland,MetteKroghb,LéonReubsaet,Three-phaseliquid-phasemicroextractionofweaklybasicdrugsfromwholeblood,[J].JournalofChromatographyB,798(2003)127–135]通過對弱堿性藥物的三相中空纖維液相微萃取的研究，表明接受相的pH應(yīng)低于藥物的pKa-3.3。相反，對于酸性物質(zhì)供相的pH值應(yīng)調(diào)節(jié)至酸性，而接受相的pH值相對較高。完成萃取之后，吸取水溶性的接受相用于HPLC和CE分析。HegeGrefslieUgland,MetteKroghb,LéonReubsaet,Three-phaseliquid-phasemicroextractionofweaklybasicdrugsfromwholeblood,[J].JournalofChromatographyB,798(2003)127–135MahnazGhambarian等[MahnazGhambarian,YadollahYamini,AliEsrafili,Three-phasehollowfibermicroextractionbasedontwoimmiscibleorganicsolventsfordeterminationoftricyclicantidepressantdrugs:Comparisonwithconventionalthree-phasehollowfibermicroextraction,[J].JournalofChromatographyA,1222(2012)5–12]建立了以第二種有機溶劑代替水溶液作為接受相的三相中空纖維液相微萃取方法。由于有機接受相與正十二烷液膜之間的能斯特擴散層較水相與正十二烷液膜之間的擴散層薄，因此有助于被測物質(zhì)從正十二烷中進入接受相。MahnazGhambarian,YadollahYamini,AliEsrafili,Three-phasehollowfibermicroextractionbasedontwoimmiscibleorganicsolventsfordeterminationoftricyclicantidepressantdrugs:Comparisonwithconventionalthree-phasehollowfibermicroextraction,[J].JournalofChromatographyA,1222(2012)5–12表2中空纖維液相微萃取在藥物分析中的應(yīng)用分析樣品被測物萃取模式分析方法供相pH值萃取溶劑參考文獻尿樣異丙嗪普馬嗪氯丙嗪三氟啦嗪兩相GC-FPD/FID9甲苯[NOTEREF_Ref395556409\h27]血漿蛇床子素兩相HPLC-FLDNG正辛醇[JunZhoua,PingZenga,ZhaoHui,etal.,ChengaApplicationofhollowfiberliquidphasemicroextractioncoupledwithhigh-performanceliquidchromatographyforthestudyoftheostholepharmacokineticsincerebralischemiahypoperfusionratplasma,[J].JournalofChromatographyB,879(2011)2304–2310JunZhoua,PingZenga,ZhaoHui,etal.,ChengaApplicationofhollowfiberliquidphasemicroextractioncoupledwithhigh-performanceliquidchromatographyforthestudyoftheostholepharmacokineticsincerebralischemiahypoperfusionratplasma,[J].JournalofChromatographyB,879(2011)2304–2310血清甲狀腺素三相LC-MS/MSNG接受相:1mol/L氨溶液中介相:甲苯[SharonYong,YizhaoChen,TongKooiLee,HianKeeLee,Determinationoftotalthyroxineinhumanserumbyhollowfiberliquid-phasemicroextractionandliquidchromatography–tandemmassspectrometry,[J].Talanta,126(2014)163-169SharonYong,YizhaoChen,TongKooiLee,HianKeeLee,Determinationoftotalthyroxineinhumanserumbyhollowfiberliquid-phasemicroextractionandliquidchromatography–tandemmassspectrometry,[J].Talanta,126(2014)163-169血漿納絡(luò)酮丁丙諾啡諾丁丙諾啡兩相UHPLC-MS/MS8.7辛醇－氯仿－甲苯（２：４：４）[WenjunSun,ShupingQu,ZhenxiaDu,Hollowfiberliquid-phasemicroextractioncombinedwithultra-highperformanceliquidchromatography–tandemmassspectrometryforthesimultaneousdeterminationofnaloxone,buprenorphineandnorbuprenorphineinhumanplasma,[J].JournalofChromatographyB,951(2014)157-163WenjunSun,ShupingQu,ZhenxiaDu,Hollowfiberliquid-phasemicroextractioncombinedwithultra-highperformanceliquidchromatography–tandemmassspectrometryforthesimultaneousdeterminationofnaloxone,buprenorphineandnorbuprenorphineinhumanplasma,[J].JournalofChromatographyB,951(2014)157-163血漿松果菊苷管花苷B類葉升麻苷異麥角甾苷兩相HPLC-UVNG正辛醇[JunZhou,QiongZhang,JiangBingSun,XiaoLiSun,PingZeng,Two-phasehollowfiberliquidphasemicroextractionbasedonmagnetofluidforsimultaneousdeterminationofEchinacoside,TubulosideB,ActeosideandIsoacteosideinratplasmaafteroraladministrationofCistanchesalsaextractbyhighperformanceliquidchromatography,[J].JournalofPharmaceuticalandBiomedicalAnalysis,94(2014)30-35JunZhou,QiongZhang,JiangBingSun,XiaoLiSun,PingZeng,Two-phasehollowfiberliquidphasemicroextractionbasedonmagnetofluidforsimultaneousdeterminationofEchinacoside,TubulosideB,ActeosideandIsoacteosideinratplasmaafteroraladministrationofCistanchesalsaextractbyhighperformanceliquidchromatography,[J].JournalofPharmaceuticalandBiomedicalAnalysis,94(2014)30-35血漿唑吡旦硝西泮阿普唑侖地西泮勞拉西泮三相HPLC-MS7.5接受相：1mol/L鹽酸溶液中介相:壬醇[NOTEREF_Ref395556533\h28]尿液血漿阿米替林丙咪嗪三甲丙咪嗪氯米帕明三相GC-MS12接受相：0.01mol/L鹽酸的甲醇溶液中介相:正十二烷[NOTEREF_Ref395556633\h29]尿液氯胺酮去甲氯胺酮脫氫去甲氯胺酮三相TFAA衍生后GC–MSNG接受相:1.0mol/L鹽酸溶液中介相:桉樹精油[AndréValledeBairros,RafaelLanaro,RafaelMenckdeAlmeida,MauricioYonamine,Determinationofketamine,norketamineanddehydronorketamineinurinebyhollow-fiberliquid-phasemicroextractionusinganessentialoilassupportedliquidmembrane,[J].ForensicScienceInternational,243,(2014)47-54AndréValledeBairros,RafaelLanaro,RafaelMenckdeAlmeida,MauricioYonamine,Determinationofketamine,norketamineanddehydronorketamineinurinebyhollow-fiberliquid-phasemicroextractionusinganessentialoilassupportedliquidmembrane,[J].ForensicScienceInternational,243,(2014)47-54血漿華法林三相HPLC–UV2.3接受相：pH=11的氫氧化鈉溶液中介相:正辛醇[MohammadrezaHadjmohammad,HodaGhambari,Three-phasehollowfiberliquidphasemicroextractionofwarfarinfromhumanplasmaanditsdeterminationbyhigh-performanceliquidchromatography,MohammadrezaHadjmohammad,HodaGhambari,Three-phasehollowfiberliquidphasemicroextractionofwarfarinfromhumanplasmaanditsdeterminationbyhigh-performanceliquidchromatography,[J].JournalofPharmaceuticalandBiomedicalAnalysis61(2012)44–49天然藥物黃酮類化合物三相HPLC-UV2接受相：pH=9.75的緩沖液中介溶劑：辛醇[MohammadrezaHadjmohammadi,HaniehKarimiyan,VahidSharifi,Hollowfibre-basedliquidphasemicroextractioncombinedwithhigh-performanceliquidchromatographyfortheanalysisofflavonoidsinEchinophoraplatylobaDC.andMenthapiperita,[J].FoodChemistry,141(2013)731-735MohammadrezaHadjmohammadi,HaniehKarimiyan,VahidSharifi,Hollowfibre-basedliquidphasemicroextractioncombinedwithhigh-performanceliquidchromatographyfortheanalysisofflavonoidsinEchinophoraplatylobaDC.andMenthapiperita,[J].FoodChemistry,141(2013)731-735注：NG－文獻中為作考察分散相液相微萃取（dispersiveliquidphasemicroextraction,DLPME）分散液相微萃取(DLPME)是自2006年后迅速發(fā)展起來的液相微萃取技術(shù)。分散相液相為萃取技術(shù)以傳統(tǒng)的液-液萃取和濁點萃取技術(shù)為基礎(chǔ)。該技術(shù)是將數(shù)微升的有機萃取溶劑通過一定的方法（如加入分散劑，或/和超聲、旋渦震蕩等）形成極其細微的小液滴，并分散到樣品水溶液中形成白色云霧狀的微乳液，對目標物質(zhì)進行萃取。待達到萃取平衡后,對乳狀液體進行離心或采用其他方法實現(xiàn)兩相分離，最后用微量注射器吸取有機相進行分析。分散相液相微萃取最初被用于分析有機化合物如多環(huán)芳烴、有機磷農(nóng)藥和氯苯化合物等[R.RahnamaKozani,Y.Assadi,F.Shemirani,M.R.MillaniHosseini,M.R.Jamali,R.RahnamaKozani,Y.Assadi,F.Shemirani,M.R.MillaniHosseini,M.R.Jamali,Talanta2007,72:387–393.分散相液相微萃取技術(shù)的基礎(chǔ)是：a.萃取溶劑能夠以微液滴的形式分散到樣品溶液中，使萃取相和水相有極大的接觸面積；b.萃取之后兩相能通過離心等手段進行相分離。與其他液相微萃取方法相比，分散相液相微萃取具有巨大的優(yōu)勢。由于萃取溶劑與水相的接觸表面積大，使得萃取平衡能迅速達到。因此，具有富集倍數(shù)高、操作時間短（一般能在5-10分鐘內(nèi)完成）、重復性好等優(yōu)點。傳統(tǒng)的分散相液相微萃取(Conventionaldispersiveliquidphasemicroextraction,Conventional-DLPME)傳統(tǒng)的分散相液相微萃取使用密度大于水的萃取溶劑，如二氯甲烷、三氯甲烷、四氯化碳、氯苯等。通過使用將萃取溶劑和分散劑快速注入到樣品的水溶液中，形成微乳狀液體使萃取溶劑與水相充分作用，實現(xiàn)被測物質(zhì)的萃取。加入分散劑的目的是促進萃取溶劑在水相中分散形成微小液滴。合適的分散劑即能與萃取溶劑又能與水互溶，能夠促進兩者之間的相互作用。但是分散劑的存在也影響了萃取之后兩相的分離。由于需要使用密度大于水的液體作為萃取溶劑，可供選擇的有機溶劑有限，而且毒性較強。同時這些溶劑密度太大、黏度較高，用微量注射器吸取時較為困難。因此出現(xiàn)了很多新的分散相液相微萃取方法，其中應(yīng)用較為廣泛的是低密度溶劑分散相液相微萃取和漂浮固體液相微萃取。低密度溶劑分散相液相微萃取（Lowdensitysolventdispersiveliquidphasemicroextraction,LDS-DLPME）顧名思義，低密度溶劑分散相液相微萃取使用的萃取溶劑的密度小于水。該方法擴大了萃取溶劑的范圍，可使用許多毒性較小的溶劑，如辛醇、甲苯、正己烷等。但是萃取之后，萃取溶劑處于水相的上層，聚集不完整，難以分離收集。因此不同文獻中先后出現(xiàn)多種萃取設(shè)備，如注射器、細口塑料瓶等。其主要作用都是將分散萃取溶劑集中便于收集。Li-ShaZhang等[Li-ShaZhang,ShuangHu,XuanChen,etal.,Anewionicliquid–water–organicsolventthreephasemicroextractionforsimultaneouspreconcentrationflavonoidsandanthraquinonesfromtraditionalChineseprescription,[J].JournalofPharmaceuticalandBiomedicalAnalysis86(2013)36Li-ShaZhang,ShuangHu,XuanChen,etal.,Anewionicliquid–water–organicsolventthreephasemicroextractionforsimultaneouspreconcentrationflavonoidsandanthraquinonesfromtraditionalChineseprescription,[J].JournalofPharmaceuticalandBiomedicalAnalysis86(2013)36-39漂浮有機液滴凝固液相微萃?。―ispersiveliquid–liquidmicroextractionbasedonthesolidificationoffloatingorganicdroplets,DLLPME-SFOD）DLLPME-SPOD與上述兩種DLLPME基本原理一致。但選用的有機萃取溶劑具有其特殊性：密度比水小且凝固點接近室溫。便于有機溶劑對分析物進行萃取并漂浮于樣品溶液表面，當溫度降低時，含有分析物質(zhì)的有機溶劑能夠凝固，實現(xiàn)相分離。因此該方法適合于高度或中等親脂性物質(zhì)的分離。常用的有機溶劑有：十一醇、１-十二醇、２-十二醇、１-溴代十六烷、正十六烷等。圖４分散相液相微萃取示意圖（Ａ）傳統(tǒng)分散相液相微萃取、（Ｂ）漂浮有機液滴凝固液相微萃取、（Ｃ）低密度溶劑分散相液相微萃取表３分散相液相微萃取在藥物分析中的應(yīng)用分析樣品被測物萃取模式分析方法供相pH值萃取溶劑分散劑參考文獻尿液7-氨基氟硝西泮ConventionalLC-ES-MS/MS含0.2M氨的溶液二氯甲烷異丙醇[MahaveerB.Melwanki,Wei-ShanChen,Hsin-YuBai,et,al.,Determinationof7-aminoflunitrazepaminurinebydispersiveliquid–liquidmicroextractionwithliquidchromatography–electrospray-tandemmassspectrometry,[J].Talanta78(2009)618–622MahaveerB.Melwanki,Wei-ShanChen,Hsin-YuBai,et,al.,Determinationof7-aminoflunitrazepaminurinebydispersiveliquid–liquidmicroextractionwithliquidchromatography–electrospray-tandemmassspectrometry,[J].Talanta78(2009)618–622尿液血漿金剛烷胺ConventionalGC-FID101,2-二溴乙烷甲醇[MirAliFarajzadeh,NinaNouri,AliAkbarAlizadehNabil,Determinationofamantadineinbiologicalfluidsusingsimultaneousderivatizationanddispersiveliquid–liquidmicroextractionfollowedbygaschromatography-flameionizationdetection,[J].JournalofChromatographyB,940(2013)142–149MirAliFarajzadeh,NinaNouri,AliAkbarAlizadehNabil,Determinationofamantadineinbiologicalfluidsusingsimultaneousderivatizationanddispersiveliquid–liquidmicroextractionfollowedbygaschromatography-flameionizationdetection,[J].JournalofChromatographyB,940(2013)142–149尿液丙咪嗪阿米替林氯米帕明地昔帕明去甲替林ConventionalGC–MS12.2四氯化碳甲醇[RieIto,MasaruUshiro,YukiTakahashi,etal.,Improvementandvalidationthemethodusingdispersiveliquid–liquidmicroextractionwithinsituderivatizationfollowedbygaschromatography–massspectrometryfordeterminationoftricyclicantidepressantsinhumanurinesamples,[J].JournalofChromatographyB,879(2011)3714–3720RieIto,MasaruUshiro,YukiTakahashi,etal.,Improvementandvalidationthemethodusingdispersiveliquid–liquidmicroextractionwithinsituderivatizationfollowedbygaschromatography–massspectrometryfordeterminationoftricyclicantidepressantsinhumanurinesamples,[J].JournalofChromatographyB,879(2011)3714–3720血漿阿米替林氯米帕明甲硫達嗪ConventionalHPLC-UV1０四氯化碳乙腈［ChaomeiXiong,JinlanRuan,YalingCai,etal.,Extractionanddeterminationofsomepsychotropicdrugsinurinesamplesusingdispersiveliquid–liquidmicroextractionfollowedbyhigh-performanceliquidchromatography,[J].JournalofPharmaceuticalandBiomedicalAnalysis49(2009)572–578ChaomeiXiong,JinlanRuan,YalingCai,etal.,Extractionanddeterminationofsomepsychotropicdrugsinurinesamplesusingdispersiveliquid–liquidmicroextractionfollowedbyhigh-performanceliquidchromatography,[J].JournalofPharmaceuticalandBiomedicalAnalysis49(2009)572–578血漿阿普唑侖溴基安定氯硝西泮勞拉西泮地西泮四氫西泮ConventionalUPLC-PDA９氯仿甲醇[PurificaciónFernPurificaciónFernándeza,CristinaGonzálezb,M.TeresaPenab，etal.,Arapidultrasound-assisteddispersiveliquid–liquidmicroextractionfollowedbyultra-performanceliquidchromatographyforthesimultaneousdeterminationofsevenbenzodiazepinesinhumanplasmasamples,[J].AnalyticaChimicaActa767(2013)88–96血漿地高辛ConventionalSALDI/MS７氯仿丙酮［Mei-ChingCheng,Kai-MingChi,SarahY.Chang,Detectionofdigoxininurinesamplesbysurface-assistedlaserdesorption/ionizationmassspectrometrywithdispersiveliquid–liquidmicroextraction,[J].Talanta115(2013)123–128Mei-ChingCheng,Kai-MingChi,SarahY.Chang,Detectionofdigoxininurinesamplesbysurface-assistedlaserdesorption/ionizationmassspectrometrywithdispersiveliquid–liquidmicroextraction,[J].Talanta115(2013)123–128水樣磺胺類喹諾酮類ConventionalUHPLC–MS/MS7.6氯仿乙腈［AntonioV.Herrera-Herrera,JavierHernández-Borges,TeresaM.Borges-Miquel,etal.,Dispersiveliquid–liquidmicroextractioncombinedwithultra-highperformanceliquidchromatographyforthesimultaneousdeterminationof25sulfonamideandquinoloneantibioticsinwatersamples,[J].JournalofPharmaceuticalandBiomedicalAnalysis75(2013)130-137chromatography;WaterAntonioV.Herrera-Herrera,JavierHernández-Borges,TeresaM.Borges-Miquel,etal.,Dispersiveliquid–liquidmicroextractioncombinedwithultra-highperformanceliquidchromatographyforthesimultaneousdeterminationof25sulfonamideandquinoloneantibioticsinwatersamples,[J].JournalofPharmaceuticalandBiomedicalAnalysis75(2013)130-137chromatography;Water血清依普沙坦纈沙坦厄貝沙坦氯沙坦替米沙坦離子流體UHPLC-MS/MS離子流體[BMIMPF6]丙酮[R.NageswaraRaoa,S.SatyanarayanaRajub,R.MastanVali,Ionic-liquidbaseddispersiveliquid–liquidmicroextractionfollowedbyhighperformanceliquidchromatographicdeterminationofanti-hypertensivesinratserum,[J].JournalofChromatographyB,931(2013)174–180]R.NageswaraRaoa,S.SatyanarayanaRajub,R.MastanVali,Ionic-liquidbaseddispersiveliquid–liquidmicroextractionfollowedbyhighperformanceliquidchromatographicdeterminationofanti-hypertensivesinratserum,[J].JournalofChromatographyB,931(2013)174–180血漿硝呋莫司芐硝唑離子流體HPLC-UV6.12離子流體[OMIM][PF6]甲醇[J.M.Padro,M.E.Marso,G.E.Mastrantonio,etal.,Developmentofanionicliquid-baseddispersiveliquid–liquidmicroextractionmethodforthedeterminationofnifurtimoxandbenznidazoleinhumanplasma,[J].Talanta107(2013)95–102]J.M.Padro,M.E.Marso,G.E.Mastrantonio,etal.,Developmentofanionicliquid-baseddispersiveliquid–liquidmicroextractionmethodforthedeterminationofnifurtimoxandbenznidazoleinhumanplasma,[J].Talanta107(2013)95–102中藥血漿尿液小檗堿巴馬亭藥根堿黃連堿離子流體熒光分析６離子流體[C8MIM][PF6]甲醇［HaoWu,Li-bingZhang,Li-mingDu,Ionicliquidsensitizedfluorescencedeterminationoffourisoquinolinealkaloids,[J].Talanta85(2011)787–793HaoWu,Li-bingZhang,Li-mingDu,Ionicliquidsensitizedfluorescencedeterminationoffourisoquinolinealkaloids,[J].Talanta85(2011)787–793中藥制劑四種黃酮化合物五種蒽醌化合物三相離子流體HPLC-UV／[C6MIM][PF6]庚醇無［NOTEREF_Ref395733411\h38］血漿西地那非伐地那非愛地那非離子流體后萃取HPLC–UV7.2離子流體［C8MIM][PF6]甲醇[ChangqinXiao,MinqiongTang,JianLi,etal.,Determinationofsildenafil,vardenafilandaildenafilinhumanplasmabydispersiveliquid–liquidmicroextraction-backextractionbasedonionicliquidandhighperformanceliquidchromatography-ultravioletdetection,[J].JournalofChromatographyB,931(2013)111–116]ChangqinXiao,MinqiongTang,JianLi,etal.,Determinationofsildenafil,vardenafilandaildenafilinhumanplasmabydispersiveliquid–liquidmicroextraction-backextractionbasedonionicliquidandhighperformanceliquidchromatography-ultravioletdetection,[J].JournalofChromatographyB,931(2013)111–116血漿華法林LDSHPLC–UV2.3辛醇甲醇[HodaGhambari,MohammadrezaHadjmohammadi,Low-densitysolvent-baseddispersiveliquid–liquidmicroextractionfollowedbyhighperformanceliquidchromatographyfordeterminationofwarfarininhumanplasma,[J].JournalofChromatographyB,899(2012)66–71HodaGhambari,MohammadrezaHadjmohammadi,Low-densitysolvent-baseddispersiveliquid–liquidmicroextractionfollowedbyhighperformanceliquidchromatographyfordeterminationofwarfarininhumanplasma,[J].JournalofChromatographyB,899(2012)66–71血漿尿液唑尼沙胺卡馬西平LDSHPLC-UV辛醇CTBA［MohammadBehbahani,FatemehNajafi,SamanBagheri,etal.,Applicationofsurfactantassisteddispersiveliquid–liquidmicroextractionasanefficientsampletreatmenttechniqueforpreconcentrationandtracedetectionofzonisamideandcarbamazepineinurineandplasmasamples,[J].JournalofChromatographyA,1308(2013)25-31MohammadBehbahani,FatemehNajafi,SamanBagheri,etal.,Applicationofsurfactantassisteddispersiveliquid–liquidmicroextractionasanefficientsampletreatmenttechniqueforpreconcentrationandtracedetectionofzonisamideandcarbamazepineinurineandplasmasamples,[J].JournalofChromatographyA,1308(2013)25-31尿液紅霉素羅紅霉素地紅霉素克拉霉素SFODLC-CAD10.8十二醇甲醇[ShaodongJia,JingLi,So-RaParka,etal.,Combinedapplicationofdispersiveliquid–liquidmicroextractionbasedonthesolidificationoffloatingorganicdropletsandchargedaerosoldetectionforthesimpleandsensitivequantificationofmacrolideantibioticsinhumanurine,[J].JournalofPharmaceuticalandBiomedicalAnalysis86(2013)204–213ShaodongJia,JingLi,So-RaParka,etal.,Combinedapplicationofdispersiveliquid–liquidmicroextractionbasedonthesolidificationoffloatingorganicdropletsandchargedaerosoldetectionforthesimpleandsensitivequantificationofmacrolideantibioticsinhumanurine,[J].JournalofPharmaceuticalandBiomedicalAnalysis86(2013)204–213尿液苯異丙胺脫氧麻黃堿SFODHPLC–UV／十一醇乙腈［TorajAhmadi-Jouibari,NazirFattahi,MojtabaShamsipur,Rapidextractionanddeterminationofamphetaminesinhumanurinesamplesusingdispersiveliquid–liquidmicroextractionandsolidificationoffloatingorganicdropfollowedbyhighperformanceliquidchromatography,[J].JournalofPharmaceuticalandBiomedicalAnalysis94(2014),145-151TorajAhmadi-Jouibari,NazirFattahi,MojtabaShamsip