荧光分光光度法测定乌饭树树叶中总黄酮含量  

Determination of Total Flavonoids in Vaccinium bracteatum Thunb Leaves by Spectrofluorimetry

作　　者：张德谨 陈义勇[2] 

机构地区：[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

出　　处：《食品工业》2014年第6期273-277,共5页The Food Industry

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434 nm,发射波长为484 nm,pH为4.4,5%AlCl3的体积为1 mL,静止时间为5 min,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=193.547 64C-2 396.382,线性相关系数r=0.997 9,线性范围为50 ng/mL^300ng/mL,方法的检出限为3.2×10-3 ng/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。According to the properties of flavonoids and aluminum ion forming a stable fluorescent complexes, Quercetin wasused as standard substance. Total flavonoids from the Vaccinium bracteatum thunb leaves were determined by fluorescencespectrophotometry. Effect of pH, standing time, aluminium chloride volume and ethanol concentration on fluorescence intensity wasinvestigated. Repeated experiment, stability experiment and recovery experiment were performed to evaluate the method. A simpleand accurate method was established to determine total ftavonoids in Vaeeinium bracteatum thub leaves. Method for determination ofthe optimal intensity of fluorescence was as follows The excitation wavelength and emission wavelength were 434 nm and 484 nmrespectively, pH was 4.4. Standing time was 5 min. Volume ofaluminium chloride （5%） was 1 mL. Ethanol concentration was 100%.Under the optimum determination conditions, quercetin concentration showed a good linear relationship with the fluorescenceintensity. The regression equation was F=193.547 64C-2 396.382 （r=0.997 9）. Linear range was 50 ng/mL300 ng/mL, thedetection limit of the method was 3.2 × 10.3 ng/mL, the average recovery and RSD were 95.43% and 5.69% respectively.Speetrophotometry is a simple, accurate and stable method, meanwhile it has good repeatability and high sensitivity,and consequently, the method can be used to determine the total flavonoids in Vaccinium brateatum thunb leaves.

关 键 词：乌饭树叶 黄酮 荧光分光光度法 

分 类 号：Q946[生物学—植物学] O657.3[理学—分析化学]