School of Materials Science and Engineering, University of Jinan, Jinan, 250022, People's Republic of China.
College of Chemistry, Chemical Engineering and Materials Science, Key Laboratory of Molecular and Nano Probes (Ministry of Education), Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Institute of Molecular and Nano Science, Shandong Normal Univeristy, Jinan, Shandong, 250014, People's Republic of China.
Mikrochim Acta. 2019 Feb 22;186(3):202. doi: 10.1007/s00604-019-3303-2.
The authors describe a fluorometric method for determination of the activity of alkaline phosphatase (ALP) and its inhibitors. Nitrogen and boron co-doped carbon dots (C-dots) with excitation/emission peaks at 490/540 nm act as the fluorescent probe. The C-dots were prepared by hydrothermal carbonization starting from 3-aminophenylboronic acid as the sole precursor. On the basis of the boronic acid-triggered specific reaction with cis-diols, the boronic acid modified C-dots can bind to ascorbic acid that is generated by ALP-catalyzed hydrolysis of ascorbic acid 2-phosphate. This results in particle aggregation and quenching of fluorescence. If the ALP inhibitor NaVO is introduced into the system, the activity of ALP is reduced and the fluorescence of C-dots recovers. This fluorometric method allows for the determination of ALP activity in the range from 0.2 to 6.0 mU mL with a detection limit of 0.16 mU mL. The IC value for the inhibitor NaVO is 3.6 μM. The method is convenient and cost-effective. It does not require complicated operations and in our perception widens the scope of applications of C-dots in bioanalytical sciences. Graphical abstract Schematic presentation of the nitrogen and boron co-doped carbon dot-based fluorometric method for determination of alkaline phosphatase (ALP) activity.
作者描述了一种用于测定碱性磷酸酶（ALP）及其抑制剂活性的荧光法。具有激发/发射峰在 490/540nm 的氮硼共掺杂碳点（C-dots）用作荧光探针。C-dots 通过以 3-氨基苯硼酸为唯一前体制备水热碳化法制备。基于硼酸与顺式二醇的特异性反应，硼酸盐修饰的 C-dots 可以与 ALP 催化水解抗坏血酸 2-磷酸生成的抗坏血酸结合。这导致颗粒聚集和荧光猝灭。如果将 ALP 抑制剂 NaVO 引入体系，则 ALP 的活性降低，C-dots 的荧光恢复。该荧光法可在 0.2 至 6.0mU mL 的范围内测定 ALP 活性，检测限为 0.16mU mL。抑制剂 NaVO 的 IC 值为 3.6μM。该方法方便且具有成本效益，不需要复杂的操作，在我们看来，它拓宽了 C-dots 在生物分析科学中的应用范围。
