Synthesis of gold and copper bimetallic nanoclusters with papain for fluorescence detection of cortisone in biological samples.

In this work, bimetallic gold and copper nanoclusters (Au-Cu NCs) were fabricated by using papain as a ligand. The as-synthesized papain-Au-Cu NCs displayed red fluorescence at 365 nm of UV lamp. The as-prepared papain-Au-Cu NCs display a strong fluorescence at 656 nm when excited at 390 nm. Papain encapsulated Au-Cu NCs exhibit distinct interaction site towards cortisone, which results red fluorescence "turn-off". Papain-Au-Cu NCs exhibited a rapid response and remarkable fluorescence quenching with the addition of cortisone as a biomarker. The developed probe is demonstrated for the quantification of cortisone by plotting the calibration graph between the fluorescence ratio (I/I) and cortisone concentration (0.031-1.00 μM) with an impressive detection limit of 1.89 nM. Interestingly, the response of papain-Au-Cu NCs towards other biomarkers and common interfering species is quite silent, signifying the selectivity of papain-Au-Cu NCs for the detection of cortisone. The probe was successfully applied to assay cortisone biomarker in urine and plasma samples. More importantly, the analytical validation of the probe was assessed by assaying cortisone in intra- and inter-day, demonstrating papain-Au-Cu NCs could serve as promising probe for the rapid detection of cortisone in biological samples.