Shandong Provincial Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu City, Shandong Province 273165, China.
Shandong Provincial Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu City, Shandong Province 273165, China; Qinghai Key Laboratory of Qinghai-Tibet Plateau Biological Resources, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining City, Qinghai Province 810001, China.
Talanta. 2017 Dec 1;175:457-462. doi: 10.1016/j.talanta.2017.07.070. Epub 2017 Jul 24.
A novel colorimetric method for the detection of tyrosinase (TYR) and its inhibitor by taking utilization of Ag-3,3',5,5'-tetramethylbenzidine (TMB) detection system has been proposed. Ag could oxidize TMB to oxidized TMB (oxTMB) and induce a blue color solution corresponding to an absorption peak centered at 652nm. The addition of dopamine (DA) could cause the reduction of oxTMB which resulted in the fading of the blue color and a decrease of the absorbance at 652nm. However, in the presence of TYR, DA could be oxidized to dopaquinone, which inhibited the reduction of oxTMB by DA, resulting in a blue color recovery and an increase of the absorbance at 652nm. Based on this finding, we propose a method to quantitatively detect TYR activity with the help of UV-vis spectroscopy. The developed assay is highly sensitive with a low detection limit of 0.010U/mL. More importantly, this method is fairly simple and inexpensive without the use of complicated nanomaterials. In addition, it constructs a useful platform for TYR inhibitor screening.
提出了一种基于 Ag-3,3',5,5'-四甲基联苯胺（TMB）检测体系利用检测酪氨酸酶（TYR）及其抑制剂的新型比色法。Ag 可以将 TMB 氧化为氧化 TMB（oxTMB），并诱导蓝色溶液，其对应于吸收峰中心在 652nm。多巴胺（DA）的加入会导致 oxTMB 的还原，从而导致蓝色褪色和 652nm 处吸光度的降低。然而，在 TYR 的存在下，DA 可以被氧化为多巴醌，这抑制了 DA 对 oxTMB 的还原，导致蓝色恢复和 652nm 处吸光度的增加。基于这一发现，我们提出了一种利用紫外-可见光谱定量检测 TYR 活性的方法。该方法具有较高的灵敏度，检测限低至 0.010U/mL。更重要的是，该方法非常简单且廉价，无需使用复杂的纳米材料。此外，它构建了一个用于 TYR 抑制剂筛选的有用平台。
