College of Chemistry & Material Science, Huaibei Normal University, Huaibei 235000, China.
Henan Key Laboratory of Biomolecular Recognition & Sensing, College of Chemistry & Chemical Engeering, Henan Joint International Research Laboratory of Chemo/Biosensing & Early Diagnosis of Major Diseases, Shangqiu Normal University, Shangqiu 476000, China.
Spectrochim Acta A Mol Biomol Spectrosc. 2022 Feb 5;266:120468. doi: 10.1016/j.saa.2021.120468. Epub 2021 Oct 6.
The photometric method is widely used in real clinical tests due to its simple operation, low cost and convenient. Many of the reported colorimetric ALP assays so far are non- ratiometric because the detection was based on changes in absorbance at a single wavelength. The development of novel colorimetric and ratiometric assay is of importance for quantitatively measuring target with high accuracy. The challenge in the design of ratiometric photometric assay is that the chromophore must have a significant spectral shift before and after binding to the target. Here, we report a colorimetric and ratiometric photometric sequential assay of AA and ALP based on the complexation between ARS and Cu and redox reaction between AA and Cu. The absorption band of ARS centered at 425 nm (yellow color), which could be shifted to 510 nm (red color) upon Cu binding. However, as far as we know, this classic color reaction has not been used to develop a ratiometric photometric method to sequentially detect AA and ALP, although photometric methods based on the regulation of other color reagents with oxidizing metal ions have been reported. The proposed sensing system shows a limit of detection for ALP at 0.24 U L and could be applied for detecting ALP in newborn calf serum. The established sensing system makes a useful contribution to the detection of ALP in complex clinical samples.
由于其操作简单、成本低、方便，光度法在实际临床测试中得到了广泛应用。到目前为止，许多报道的比色 ALP 测定法都是非比率的，因为检测是基于在单个波长处的吸光度变化。新型比色和比率测定法的发展对于定量测量具有高精度的目标非常重要。比率光度测定法设计的挑战在于，生色团在与靶标结合前后必须有明显的光谱位移。在这里，我们报告了一种基于 ARS 和 Cu 的络合以及 AA 和 Cu 之间的氧化还原反应的 AA 和 ALP 的比色和比率光度顺序测定法。ARS 的吸收带位于 425nm（黄色），结合 Cu 后可转移到 510nm（红色）。然而，据我们所知，尽管已经报道了基于调节其他带有氧化金属离子的显色试剂的光度法，但这种经典的显色反应尚未用于开发比率光度法来顺序检测 AA 和 ALP。所提出的传感系统对 ALP 的检测限为 0.24 U L，可用于检测新生小牛血清中的 ALP。所建立的传感系统为复杂临床样本中 ALP 的检测做出了有益的贡献。
