Microchimica Acta ( IF 5.3 ) Pub Date : 2021-08-02 , DOI: 10.1007/s00604-021-04951-6 Fanpeng Ran 1 , Chen Ma 1 , Yueci Xiang 1 , Yixuan Xu 1 , Xiaoyan Liu 1 , Haixia Zhang 1
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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). Au3+, 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.
Graphical abstract
中文翻译：
基于酸性磷酸酶触发阻断内部过滤效应的荧光和比色双通道传感器
基于氧化的 3,3',5,5'-四甲基联苯胺 (oxTMB) 和罗丹明 B (RB) 之间的内部过滤效应，构建了酸性磷酸酶 (ACP) 活性的比色和荧光双通道检测方法。Au 3+原位形成金纳米颗粒 (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的发射和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促进了2-磷酸-l的水解。-抗坏血酸三钠盐（AAP）生成抗坏血酸（AA），可以抑制RB和oxTMB之间的内部过滤作用。测定 ACP 无需准备材料。比色法和荧光法可以分别在 0.06–5.0 mU/mL 和 0.03–5.0 mU/mL 范围内量化 ACP。此外，还构建了智能手机辅助传感平台，用于现场监测0.75-50 mU/mL范围内的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。
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