The development of a novel AND logic based fluorescence probe for the detection of peroxynitrite and GSH.

We have developed a novel AND logic based fluorescence probe for the simultaneous detection of ONOO- and GSH ( GSH-PF3 ). The GSH-PF3 probe was synthesised over three steps starting from commercially available fluorescein. The probe was constructed by attaching the GSH reactive motif, 2,4-dinitrobenzenesulfonyl, to the previously reported boronate fluorescence based probe, PF3 . GSH-PF3 produced only a small fluorescence response towards the addition of GSH or ONOO- separately. However, when the probe was exposed to both analytes, there was a significant (40-fold) fluorescence enhancement. GSH-PF3 demonstrated an excellent selectivity towards both GSH and ONOO- . In cellular imaging experiments the probe was shown to be cell permeable with no 'turn-on' response observed for the addition of either GSH or ONOO- separately. However, in the presence of both analytes, a clear fluorescence response was observed in live cells. GSH-PF3 was further able to monitor the co-existence of metabolically produced ONOO- and GSH by exogenous stimulation.