Fluorescence detection of protamine, heparin and heparinase II based on a novel AIE molecule with four carboxyl.

In this research, a new DSA (Distyryl-anthracene) derivative with four carboxyl groups was designed and synthesized. This molecule exhibits aggregation-induced emission property (AIE). With the AIE character, a convenient and sensitive fluorescent probe for the detection of protamine, heparin and heparinase has been developed. The protamine can directly induce the aggregation of probe, which caused by electrostatic attraction. In this way, the turn-on detection of protamine is achieved, and the detection limit is as low as 30 ng mL. When heparin appears, the probes will be redisperse in solution, which causes a decrease in fluorescence intensity. Besides, this method also shows good selectivity and sensitivity, and the linear range of heparin is from 0.08 to 8 μg mL with detection limit of 37 ng mL. After hydrolyzing heparin by heparinase, the probes rebind with protamine and the fluorescence enhance. The fluorescence enhancement was linearly related to the concentration of heparinase in the range of 0.02-2.0 μg mL and detection limit as low as 143.7 ng mL. In addition, the results exhibited that the recovery percentage of heparinase in bovine samples reached to 96-101%.