微囊藻毒素-LR阻抗型免疫電化學(xué)檢測(cè)方法的研究.pdf

1、江南大學(xué)碩士學(xué)位論文微囊藻毒素LR阻抗型免疫電化學(xué)檢測(cè)方法的研究姓名：石慧申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：營(yíng)養(yǎng)與食品衛(wèi)生學(xué)指導(dǎo)教師：王洪新20090701AbstractAbstractRecently，withtheoccurrenceofwidespreadheavycyanobacterialblooms，severalseriouswaterandfoodsafetyepisodesinvolvingmicrocystinsisattr

2、actingglobalattentionMicrocystin—LR(MCLR)jsthemostcommonandtoxicinallcyanophyceantoxinExistinganalyticalmethodsforMCLRhavemanydisadventagessuch舔highcost，tedioussamplepreparationandlowefficiencyTherefore，thedevelopmentofa

3、highlysensitiveandrapiddetectionmethodforMCLRisofsignificanceInthispaperthroughthepreparationofimmuneantigenandspecificantibodiestoMC—LR，animmunosensorwhichbaSedonimmunoassaytechniqueandselfaSsemblednanomaterialWaSdevelo

4、pedforthedeterminationofMCLRThemainpointsofthisdissertationaresummarizedasfollows：Theimmunoanddetectionantigenwerepreparedusingthemethodsof“activeester’’and“EDCcatalyze”HighselectivitypolyclonalantibodyagainstMCLRWasobta

5、inedfromtheNewZealandrabbitimmunizedwiththeimmunoantigenTheantiserumwastestedbyindirectedELISAThetiterofthisantibodyishigherthan52x10珥afterpurification；111ecalibrationcurvebyindirectedELISAshowsthattherangeofquantitative

6、detectionWaS025“4pg／L麗廿1artIC50valueof068“g／L11lccrossreactivitywithMCRRwas219％，andwerealllowerthanl％謝tllMCLWandMC—LFAlabelfreeimmunosensorfordetectionofMCLRhasbeendevelopedbyimmobilizingantiMCLRonNanoAu／Lcysteinecoatede

7、lectrodeUsingthedevelopedmethodMCLRcouldbedeterminedwithadetectionlimitof182x10。2ng／mL(S／N=3)andlinearitybetweenO05and3000nedmLThewholedetectingtimewasonly15min111eaveragerecoveryrateWasintherangeof968～1125％11leimmunosen

8、sorexhibitedlong—termstabilityandgoodreproducibilityofthesignalcouldbeobtainedupto42timesbyCV，nlecoefficientofvariationWas358％；Furthermore，goodstabilityin30daysat4℃Immobilizationofanti—MCLRhasbeenbasedonselfassembletechn

9、iqueof1，6一hexanedithiolandNanoAu11圮resultingimmunosensordisplayedahighsensitivityandawidelinearrangeforthedetectionofMCLRandrespondedtotheMC—LRintherangeof025～950ng／mLwithalowdetectionlimitof85x10之rig／mE(S／N=3)Moreoverth

10、erelativestandarddeviationforeightparalleldeterminationsis748％andgoodstabilityin30daysat4℃111eaveragerecoveringWasintherangeof926％。1183％11ledetetiontimeWaS15minImmobilizingantiMCLRonmultiwalledcarbonnanotubes(MWNTsyroomt

11、emperatureionicliquids(RTILs)coatedelectrode，animmunosensorfordetectionofMCLRWasdevelopedMCLRcouldbedeterminedwithalowerdetectionlimitof17x103ng／mL(S／N=3)andlinearitybetween5x10jng／mLand1xl0zng／mLneaveragerecoverywasinth

12、erangeof8095％12723％SomecommonionsinwaterhavenointerferencewitllMCLRInviewofstorageconditionsofimmunosensors，asolutionwhichcanprotectelectrodesisfirstputforwardThemodifiedelectrodeswerestoredinapH74phosphatebuffersolution