College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu Shandong, 273165, PR China.
College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu Shandong, 273165, PR China.
Talanta. 2020 May 15;212:120768. doi: 10.1016/j.talanta.2020.120768. Epub 2020 Jan 21.
This study demonstrates a novel and convenient ratiometric fluorescent method for the detection of alkaline phosphatase (ALP) activity. Amino-functionalized mesoporous silica nanoparticle-gold nanoclusters (MSN-AuNCs) nanocomposites were integrated with o-phenylenediamine (OPD) to form a ratiometric fluorescence nanoplatform. The presence of ALP induced the generation of quinoxaline (QX) derivative which called 3-(dihydroxyethyl)furo[3,4-b]quinoxaline-1-one (DFQ) with strong fluorescence emission at 450 nm, while the orange-red fluorescence of MSN-AuNCs at 580 nm was slightly quenched. Meanwhile, an obvious fluorescence color change from orange-red to purple and finally to blue can be observed by naked eyes with the increasing of ALP concentration. Therefore, employing the fluorescence emission of DFQ at 450 nm as the reporter signal and the fluorescence emission of MSN-AuNCs at 580 nm as a reference signal, a sensitive ratiometric detection method for ALP was developed. Quantitative detection of ALP activity in the linear range from 0.2 to 80 U/L with a detection limit of 0.1 U/L can be realized in this way, which endows the assay with high sensitivity enough for practical detection of ALP in human serum samples.
本研究展示了一种新颖、简便的比率荧光法用于检测碱性磷酸酶（ALP）活性。氨基功能化介孔硅纳米粒子-金纳米簇（MSN-AuNCs）纳米复合材料与邻苯二胺（OPD）结合形成比率荧光纳米平台。ALP 的存在诱导了喹喔啉（QX）衍生物的生成，称为 3-（二羟乙基）呋喃[3,4-b]喹喔啉-1-酮（DFQ），其在 450nm 处具有强荧光发射，而 MSN-AuNCs 的橙红色荧光在 580nm 处略有猝灭。同时，随着 ALP 浓度的增加，可以通过肉眼观察到明显的荧光颜色从橙红色变为紫色，最后变为蓝色。因此，采用 450nm 处的 DFQ 荧光发射作为报告信号，580nm 处的 MSN-AuNCs 荧光发射作为参考信号，开发了一种用于 ALP 的灵敏比率检测方法。通过这种方法，可以实现 ALP 活性在 0.2 至 80 U/L 的线性范围内的定量检测，检测限为 0.1 U/L，这使得该测定法具有足够的高灵敏度，可用于实际检测人血清样品中的 ALP。
