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1、基于發(fā)光金納米粒子熒光增強(qiáng)法測定溶菌酶摘要參照?SymbolmAs增加到8.96 ?SymbolmAs,這進(jìn)一步說明溶菌酶和GNPs之間的靜電作用可能使得GNPs表面鈍化,從而導(dǎo)致GNPs的熒光增強(qiáng)。 圖4離子強(qiáng)度對體系熒光增敏的影響?yīng)ぁ ig.4EffectofNaClonthefluorescenceenhancementofGNPsat0.01mol/LpH7.0PBSbuffer ConcentrationsoflysozymeandGNPses;10-6and4.010-6mol/L,respectively.
2、圖5熒光衰減曲線 Fig.5Fluorescencedecaycurves λex=320nm,λem=410nm. 3.3實(shí)驗(yàn)條件優(yōu)化 3.3.1發(fā)光金納米粒子濃度選擇實(shí)驗(yàn)表明,GNPs的熒光強(qiáng)度隨著其濃度的增加而增大。但GNPs濃度過大(>5.010-5mol/L,依據(jù)BSA濃度計(jì)算)時(shí),會(huì)出現(xiàn)自猝滅現(xiàn)象,熒光強(qiáng)度反而降低。控制較低的GNPs的濃度,能降低熒光本底,對提高測定靈敏度有利。但過低的GNPs濃度,會(huì)導(dǎo)致發(fā)光不穩(wěn)定,也減小了測定溶菌酶的線性范圍。本實(shí)驗(yàn)
3、GNPs濃度選擇4.010-6mol/L。 3.3.2酸度與介質(zhì)的影響圖6給出在4.010-6mol/L溶菌酶存在和不存在的條件下,pH值對發(fā)光金納米粒子熒光強(qiáng)度的影響。圖6在溶菌酶存在(F)和不存在(F0)條件下,pH值對發(fā)光金粒子熒光強(qiáng)度的影響?yīng)ぁ ig.6EffectofpHonfluorescenceintensityofGNPsinthepresence(F)andabsence(F0)oflysozyme GNPs和溶菌酶濃度均為4.010-6mol/L(Boththeconcentrationoflysoz
4、ymeandGNPses;10-6mol/L)。發(fā)光金粒子本身在不同pH值熒光強(qiáng)度呈現(xiàn)一定的波動(dòng),在pH>11時(shí)強(qiáng)度較大,這與 13MEiF,HeXA,SchaaffTG,,ZhouDB,HaoJY,LuYH,ChenSM.SpectrochimicaActaPartA,2009,71(5):1795~1798 17HeH,XieC,RenJC.Anal.Chem.,2008,80(15):5951~5957 18LIUMei-Gui,CAOChun,CAOMing,ZHUChang-Qing(劉玫瑰,曹春,曹明,朱昌青).Chines
5、eJ.Anal.Chem.(分析化學(xué)),2009,37(10):1503~1506 19LiuL,ZhengHZ,ZhangZJ,HuangYM,ChenSM,HuYF.SpectrochimicaActaPartA,2008,69(3):701~705 20ZhengJ,PettyJT,DicksonRM.J.Am.Chem.Soc.,2003,125(26):7780~7781 21MohamedMB,VolkovV,LinkS,El-SayedMA.Chem.Phys.Lett.,2000,317(6):517~523
6、22BigioniTP,ethodforDeterminationofLysozymeUsing FluorescentGoldNanoparticlesasProbe QIANZhang-Sheng,LIUMei-Gui,TIANDa-Hui,HAODan,ZHUChang-Qing (AnhuiKeyLaboratoryofChemo-Biosensing,CollegeofChemicalandMaterialsScience, AnhuiNormalUniversity,ehasthepropertyofdissolving
7、somebacteria,itcanbeappliedinmedicaltreatment,biologicalengineering,andespeciallyinfoodantisepsistoreplacechemicallysynthesizedones.Therefore,thedevelopmentofasimpleanalyticalmethodforlysozymeassayisveryimportant.Here,odifiedfluorescentgoldnanoparticles(GNPs)eethodforthedeterminatio
8、noflysozymehasbeend