荧光分光光度法测定乌饭树树叶中总黄酮含量.docx

荧光分光光度法测定乌饭树树叶中总黄酮含量张德谨1,2，陈义勇2*1. 中国矿业大学化工学院（徐州 221008）；2. 常熟理工学院生物与食品工程学院（常熟 215500）摘 要 根据黄酮类化合物与Al3+之间能够形成稳定荧光络合物的性质, 以槲皮素作为标准物, 采用荧光分光光度法检测乌饭树树叶中总黄酮的含量,探讨pH、静止时间、氯化铝体积、乙醇体积分数对荧光强度的影响,确定测定荧光强度的条件。利用重复性试验、稳定性试验以及加标回收率试验对该检测方法进行评价,建立一种检测乌饭树树叶中总黄酮含量的荧光分光光度法。确定的最佳测定荧光强度的方法:激发波长为434nm,发射波长为484nm,pH为4.4,5%AlCl3的体积为1mL,静止时间为5min,采用无水乙醇定容。在最佳测定条件下,槲皮素浓度与荧光强度呈良好的线性关系,线性回归方程为F=193.54764C-2396.382，线性相关系数r=0.9979,线性范围为50ng/mL～300ng/mL,方法的检出限为3.2×10-3ng/mL,平均回收率为95.43%,相对标准偏差RSD(n=9)为5.69%。荧光分光光度法简单、准确、稳定, 重复性好, 灵敏度高, 可用于乌饭树树叶中总黄酮定量检测。DeterminationofTotalFlavonoidsinVacciniumbracteatumThunbLeavesbySpectrofluorimetry关键词乌饭树叶;黄酮;荧光分光光度法ZhangDe-jin1,2,ChenYi-yong2*SchoolofChemicalEngineeringandTechnology, ChinaUniversityofMiningandTechnology(Xuzhou221008);SchoolofBiologyandFoodEngineering,ChangshuInstituteofTechnology(Changshu215500)Abstract According to the properties of flavonoids and aluminum ion forming a stable fluorescent complexes, Quercetinwasused as standard substance. Total fl avonoids from the Vaccinium bracteatum thunb leaves were determined by fluorescence spectrophotometry.EffectofpH,standingtime,aluminiumchloridevolumeandethanolconcentrationonfluorescenceintensitywas investigated.Repeatedexperiment,stabilityexperimentandrecoveryexperimentwereperformedtoevaluatethemethod.AsimpleandaccuratemethodwasestablishedtodeterminetotalflavonoidsinVacciniumbracteatumthubleaves.Methodfordeterminationoftheoptimalintensityoffluorescencewasasfollows:Theexcitationwavelengthandemissionwavelengthwere434nmand484nmrespectively.pHwas4.4.Standingtimewas5min.Volumeofaluminiumchloride(5%)was1mL.Ethanolconcentrationwas100%.Under the optimum determination conditions, quercetin concentration showed a good linear relationship with the fluorescenceintensity.TheregressionequationwasF=193.54764C-2396.382(r=0.9979).Linearrangewas50ng/mL~300ng/mL,thedetectionlimitofthemethodwas3.2×10-3ng/mL,theaveragerecoveryandRSDwere95.43%and5.69%respectively.Spectrophotometryisasimple,accurateandstablemethod,mea