Development of luminol-fluorescamine-PVP chemiluminescence system and its application to sensitive tyrosinase determination

Fluorescamine is a popular fluorescent probe. We report for the first time that luminol chemiluminescence (CL) can be enhanced by fluorescamine in the presence of PVP. The CL intensity of luminol-fluorescamine-PVP is about 26 times stronger than that of luminol. Both the removal of oxygen and the addition of superoxide dismutase (SOD) decrease CL intensity, thiourea and NaN3 have little effect on CL intensities, indicating that O-2(center dot-) is critical for CL. Interestingly, o-quinone generated from phenol by tyrosinase obviously inhibited the CL intensity. Inspired by such quenching effect on the luminol-fluorescamine-PVP CL system, a sensitive CL sensing for the determination of tyrosinase activity was developed. The method can detect tyrosinase in the range of 0.07-1.5 mu g mL(-1) (0.19-4.02 U mL(-1)) with the detection limit of 0.035 mu g mL(-1) (0.094 U mL(-1)). Moreover, this method exhibits satisfied recoveries for the spiked human serum samples.