Wang Yang, Wang Wei, Fang Yuan, Tao Dengwu, Tang Kaiyan, Nie Jinshan, Chang Baisong
Suzhou Chien-Shiung Institute of Technology, No. 1 Chien-Shiung Road, New Town of Science and Education, Taicang, Jiangsu, 215411, CHINA.
Nhwa Pharma. Corporation, No.18, Yangshan Road, Economic Development Zone, Xuzhou, 221000, CHINA.
Nanotechnology. 2024 Oct 10. doi: 10.1088/1361-6528/ad8582.
Accurate and point-of-care cholesterol detection is of paramount significance for the prevention of cardiovascular diseases. The colorimetric assay based on peroxidase is a commonly used approach for cholesterol detection, without requiring any complicated biomolecular labeling or sophisticated instrumentation. Copper nanoclusters (CuNCs), exhibiting luminescent properties and peroxidase activity, have garnered significant attention in biomedical application recently. Herein, the glutathione-stabilized copper nanoclusters (GSH-CuNCs) were prepared with an easy one-pot method, employing glutathione as both a reducing agent and stabilizer. An optimization of the GSH-CuNCs preparation was carried out to obtain the highest peroxidase-like activity. UV-Vis absorption was measured to explore the steady-state kinetics of the GSH-CuNCs-catalyzed oxidation of 3,3',5,5' - tetramethylbenzidine (TMB) by H2O2. A colorimetric method for cholesterol detection was developed by combining the catalytic reaction of CuNCs and the enzymatic oxidation of cholesterol with cholesterol oxidase (ChOx). Under the optimized conditions, the UV-Vis absorbance of oxidized TMB (oxTMB) is proportional to the concentration of cholesterol within the range of 6.2-187.5 μM, and the limit of detection (LOD) is determined to be 3.0 μM. More importantly, cholesterol levels can be directly distinguished with the naked eye. Furthermore, the practicality of the method for detecting cholesterol in human serum has been verified with promising results. As expected, this simple, cost-effective, and easy-to-operate colorimetric method for cholesterol detection has potential applications in clinical diagnosis and provides valuable insights into the colorimetric sensing based on CuNCs.
准确且即时的胆固醇检测对于预防心血管疾病至关重要。基于过氧化物酶的比色测定法是一种常用的胆固醇检测方法，无需任何复杂的生物分子标记或精密仪器。具有发光特性和过氧化物酶活性的铜纳米簇（CuNCs）最近在生物医学应用中受到了广泛关注。在此，采用简单的一锅法制备了谷胱甘肽稳定的铜纳米簇（GSH-CuNCs），使用谷胱甘肽作为还原剂和稳定剂。对GSH-CuNCs的制备进行了优化，以获得最高的类过氧化物酶活性。通过紫外可见吸收光谱测量来探究GSH-CuNCs催化H2O2氧化3,3',5,5'-四甲基联苯胺（TMB）的稳态动力学。通过将CuNCs的催化反应与胆固醇氧化酶（ChOx）对胆固醇的酶促氧化相结合，开发了一种胆固醇比色检测方法。在优化条件下，氧化TMB（oxTMB）的紫外可见吸光度在6.2 - 187.5 μM范围内与胆固醇浓度成正比，检测限（LOD）确定为3.0 μM。更重要的是，胆固醇水平可以直接用肉眼区分。此外，该方法检测人血清中胆固醇的实用性已得到验证，结果令人满意。正如预期的那样，这种简单、经济高效且易于操作的胆固醇比色检测方法在临床诊断中具有潜在应用，并为基于CuNCs的比色传感提供了有价值的见解。
