研究目的
To develop a rapid and sensitive fluorometric assay for the determination of alkaline phosphatase (ALP) activity based on the aggregation-induced quenching of the fluorescence of copper nanoclusters (CuNCs).
研究成果
The developed fluorometric assay based on CuNCs and the inner filter effect provides a sensitive, selective, and rapid method for the determination of ALP activity. The method has potential applications in biomedical research and diagnostics.
研究不足
The study may have limitations in terms of the specificity of the fluorescence quenching mechanism and potential interference from other substances in complex biological samples.
1：Experimental Design and Method Selection:
The study utilized L-histidine as the stabilizer and ascorbic acid (AA) as the reductant for the synthesis of CuNCs. The fluorescence quenching effect of PNP on CuNCs was exploited to develop a fluorometric assay for ALP activity.
2：Sample Selection and Data Sources:
The experiments involved the use of CuNCs, PNPP, and ALP in phosphate buffer solutions. Human serum samples were also tested for ALP activity.
3：List of Experimental Equipment and Materials:
Instruments included a LS55 fluorescence spectrometer, a spectrum one FTIR spectrophotometer, a TecnaiG20 transmission electron microscope, and a Lambda 35 UV analyzer. Chemicals included CuCl2·2H2O, L-Histidine, AA, PNPP, PNP, and ALP.
4：Experimental Procedures and Operational Workflow:
CuNCs were synthesized and characterized. The fluorescence quenching effect of PNP on CuNCs was studied. ALP activity was determined by measuring the fluorescence intensity changes in the presence of PNPP and ALP.
5：Data Analysis Methods:
The relationship between fluorescence intensity and ALP concentration was analyzed to determine the linear range and detection limit of the assay.
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