多肽荧光探针HWEHH对水中铜离子的高通量检测

High-throughput detection of copper ions in water by polypeptide probe HWEHH

目的:开发组氨酸—色氨酸—谷氨酸—组氨酸—组氨酸(HWEHH)水溶性荧光探针结合酶标仪实现水中Cu^(2+)的高通量检测。方法:通过选择性试验研究荧光探针结合Cu^(2+)的特异性;滴定试验确定探针检测Cu^(2+)的线性范围以及检测限,样品加标回收探究探针检测水中Cu^(2+)的准确度。结果:多肽探针溶液浓度为2μmol/L时,其检测Cu^(2+)的线性范围为0~1.5μmol/L,线性方程Y=-1135.0857 X+2048.1876,络合常数为1.3×106 mol/L,检测限为0.036μmol/L,高通量检测的线性范围为0~1.5μmol/L,加标回收率为93.6%~101.9%。结论:HWEHH多肽荧光探针可以实现水溶液中Cu^(2+)的高通量检测。

Objective:A high-throughput detection method for Cu^(2+) was developed with a well designed water-soluble fluorescent probe(HWEHH)and the microplate reader.Methods:The specificity of the fluorescent probe for Cu^(2+) determination was studied with the selectivity experiment.The linear range and detection limit of the probe for Cu^(2+) determination was detected with titration experiment,and the accuracy of the probe for the detection of Cu^(2+) in water was studied with sample spiked recovery method.Results:The fluorescent probe was able to specifically detect Cu^(2+).When the concentration of peptide probe was 2μmol/L,the linear range of Cu^(2+) detection was 0~1.5μmol/L.The linear equation was Y=-1135.0857 X+2048.1876,and the detection limit was 0.036μmol/L.The binding constant of HWEHH-Cu^(2+) complex was 1.3×106 mol/L.The linear range of high-throughput detection method was 0~1.5μmol/L,and the recovery rate was 93.6%~101.9%.Conclusion:HWEHH peptide fluorescent probe can be used for high-throughput detection of Cu^(2+)in aqueous solution.