Polydopamine coated copper nanoclusters with aggregation-induced emission for fluorometric determination of phosphate ion and acid phosphatase activity

The preparation of aggregation-induced emission-type copper nanoclusters (CuNCs) capped with polydopamine (PDA) is described. PDA was formed via in situ polymerization of dopamine in the presence of alkaline polyethylenimine. The PDA-capped CuNCs (PDA-CuNCs) exhibit orange fluorescence with maximal emission at 580 nm upon excitation at 340 nm, a storage stability of at least 2 weeks, and a quantum yield (QY) of 2.54% in aqueous solution. The QY is 28-fold higher than that of sole CuNCs. The fluorescence of the PDA-CuNCs is quenched by Fe3+ ion while it is recovered by PO43- due to its stronger affinity for Fe3+. On this basis, a fluorometric phosphate assay was developed that has a 1.5 nM detection limit and a linear range over 0.003-70 mu M. The method was satisfactorily applied to the determination of phosphate in local tap water and human sera, and the results agreed well with those obtained by a colorimetric method. In the presence of acid phosphatase (ACP), PO43- is produced by the catalytic hydrolysis of adenosine triphosphate (ACP substrate). Thus, a fluorogenic assay for screening ACP activity was established. Response is linear over the activity range 0.0012-25 U L-1, with a detection limit of 0.001 U L-1 (at S/N=3).