基于胶体金免疫层析技术制备快速检测中药中黄曲霉毒素b1试纸条的.研究

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1、...都中药大学2012硕士学论文AbstractObjective:TodevelopthecolloidalgoldteststripbasedonthelateralflowimmunechromatographytechniqueforrapiddetectionofaflatoxinB1inlotusseed,angelicasinensis,gingerpowder,pericarpiumcitrireticulatae,panaxquinquefoliumandradixpae

2、oniaealba.TheestablishedmethodfordetectingaflatoxinB1wassimple,fast,economicandefficient.Methods:TheantibodyofAFB1wasevaluatedtiterbyusingenzyme-linkedimmunosorbentassay(ELLISA);Nanogoldparticles(NGP)werepreparedforusingthemethodofcitricacidthreesodi

3、umreduction,colloidalgoldsolutionwasusedtoidentifythemorphologybyocularestimate,observedistributionofapparentbythetransmissionelectronmicroscopeanddeterminatethemaximumabsorptionwavelengthbyUVSpectrophotometry;Theindirectcompetitivecolloidalgoldimmune

4、chromatographytechnology(GICA)wasapplytooptimizeofthecouplingconcentrationofcolloidalgoldlabeledwithmonoclonalantibody,thesolutionpHandtheconcentrationofAFB1-BSA(Textline)andGoatanti-MouseIgG(Controlline);Thepreparationofthecolloidalgoldstripwasprelim

5、inarytestedthecontentofAFB1anddeterminetheminimumdetectionlimit.Theperformanceofthestripwasinspected(includedspecificity,stabilityandreproducibility).ThestripmethodwasusedtodetectedAFB1indifferentbatchsamplesof6kindsoftraditionalChinesemedicine(TCM)an

6、dUFLC-MS/MSmethodwaspreparedtodeterminatedinthecontentofAFB1inTCM.Results:Theresultsofpreparationstrip:Themonoclonalantibodies(McAb)ofAFB1weredeterminedwithcoatingantigenandtheresultsindicatedthecontentofMcAbwas3.8mg/mL.ThetiteroftheMcAbwas1:1.0×104an

7、dthe50%inhibitionconcentrationwas0.01mg/mL.Goldparticlesdiameterwasdeteminatedabout40nmbyTEM.Thecolloidalgoldsolutiongetmaximumabsorptionwavelengthat518nm.ThecouplingconcentrationofcolloidalgoldlabeledwithMcAbwas6µg/mL,undertheconditionofpH8.2andcolloi

8、dalgoldsolutionwascoatedwithMcAbwereobtainedthemaximumabsorbance.TheconcentrationofAFB1-BSA(Textline)andIII......万方数据......都中药大学2012硕士学论文Goatanti-MouseIgG(Controlline)were0.3mg/mLand0.5mg/mL,respectively.Thelimitofdetectionofthecons