State Key Laboratory of Applied Organic Chemistry, College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou, 730000, China.
Mikrochim Acta. 2021 Aug 2;188(8):282. doi: 10.1007/s00604-021-04951-6.
A colorimetric and fluorescent dual-channel detection method for acid phosphatase (ACP) activity has been constructed, based on the internal filtering effect between oxidized 3,3',5,5'-tetramethylbenzidine (oxTMB) and rhodamine B (RB). Au, which in situ form gold nanoparticles (AuNPs), can oxidize colorless 3,3',5,5'-tetramethylbenzidine (TMB) to oxTMB (blue color). The fluorescence of RB can be quenched by oxTMB due to the spectral overlap of emission of RB and absorption of oxTMB. By means of the above process, ACP can be determined because ACP promotes the hydrolysis of 2-phospho-L-ascorbic acid trisodium salt (AAP) to generate ascorbic acid (AA), which can inhibit the internal filtering effect between RB and oxTMB. No material preparation was needed for the determination of ACP. The colorimetric and fluorimetric methods can quantify ACP in the range 0.06-5.0 mU/mL and 0.03-5.0 mU/mL, respectively. Furthermore, a smartphone-assisted sensing platform has been constructed for on-site monitoring of ACP in the range 0.75-50 mU/mL, and the detection limit is 0.3 mU/mL. The methods developed can measure ACP in human serum successfully.
构建了一种基于氧化 3,3',5,5'-四甲基联苯胺（oxTMB）与若丹明 B（RB）之间的内滤效应的酸性磷酸酶（ACP）活性的比色和荧光双通道检测方法。金原位形成金纳米粒子（AuNPs），可以将无色的 3,3',5,5'-四甲基联苯胺（TMB）氧化为 oxTMB（蓝色）。由于 RB 的发射光谱与 oxTMB 的吸收光谱重叠，RB 的荧光可以被 oxTMB 猝灭。通过上述过程，可以测定 ACP，因为 ACP 可以促进 2-磷酸-L-抗坏血酸钠三钠盐（AAP）的水解，生成抗坏血酸（AA），从而抑制 RB 和 oxTMB 之间的内滤效应。该测定方法不需要进行材料制备。比色法和荧光法分别可以在 0.06-5.0 mU/mL 和 0.03-5.0 mU/mL 的范围内定量 ACP。此外，构建了一个智能手机辅助传感平台，用于在 0.75-50 mU/mL 的范围内现场监测 ACP，检测限为 0.3 mU/mL。所开发的方法可以成功地测量人血清中的 ACP。
