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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 that flavonoids can form stable fluorescent complex with Al3 +, using quercetin as standard, the content of total flavonoids in leaves of M. officinalis L. was detected by fluorescence spectrophotometry. The effects of pH, rest time, chlorination Aluminum volume, volume fraction of ethanol on the fluorescence intensity, determine the determination of fluorescence intensity conditions. The repeatability test, stability test and spike recovery test were used to evaluate the detection method, and a fluorescence spectrophotometry method was established for the determination of total flavonoids in the leaves of Ulva pertussis. The best determination of the fluorescence intensity method: excitation wavelength of 434 nm, emission wavelength of 484 nm, pH of 4.4, 5% AlCl3 volume of 1 mL, the rest time of 5 min, with ethanol volume. Under the optimum conditions, the linear relationship between quercetin concentration and fluorescence intensity was linear, the linear regression equation was F = 193.547 64C-2 396.382, the linear correlation coefficient was 0.997 9, the linear range was 50 ng / mL to 300 ng / mL. The detection limit was 3.2 × 10-3 ng / mL. The average recovery was 95.43%. The relative standard deviation (RSD) was 5.69%. Fluorescence spectrophotometry is simple, accurate, stable, reproducible, and highly sensitive. It can be used for the quantitative determination of total flavonoids in the leaves of Ulva rubra.