Chemoselective and quantitative detection of acrolein based on pyrene excimer fluorescence via cycloaddition reaction of conjugated imine

Acrolein is a highly reactive and toxic aldehyde generated in various biological and environmental contexts, including tobacco combustion and cellular oxidative stress. Herein, we present a novel quantitative method for acrolein detection using Probe A, which features an ethanolamine skeleton and a pyrene moiety. This method employs a 4 + 4 dimerization reaction, enabling selective interaction with acrolein, resulting in excimer fluorescence emission proportional to the acrolein concentration. This approach offers high sensitivity and selectivity across a broad dynamic range from micromolar to millimolar concentrations. Optimization of the reaction and measurement solvents further improved the detection sensitivity. This protocol, which incorporates chloroform extraction and a direct ethanol reaction, allows for reliable quantification of acrolein in complex biological matrices without contaminant interference. The application of this method enabled the accurate measurement of acrolein produced by tobacco combustion and by living cells subjected to oxidative stress, including the quantification of hydrogen peroxide concentration-dependent acrolein production. This method overcomes the key limitations of conventional acrolein detection methods by providing a simple, sensitive, and versatile platform suitable for diverse sample types, including tobacco smoke and living cells.