This study reports the synthesis of gold nanoparticles (AuNPs) using an unconventional, eco-friendly method that utilizes cinnamon bark extract as a reducing agent and stabilizer. The synthesized AuNPs were characterized using UV–Vis spectroscopy and transmission electron microscopy (TEM). UV–Vis spectrum shows the surface plasmon resonance (SPR) peak around 535 nm. TEM shows the spherical shape of AuNPs and the particle size distribution at around 35 nm. The ability of the synthesized AuNPs as a quencher of eosin Y dye was monitored using a fluorometer. It was found that in the presence of AuNPs, the fluorescence peak of eosin is quenched. The fluorescence of the mixture of eosin and AuNPs is enhanced in the presence of bovine serum albumin (BSA) protein. TEM shows that the average size of the AuNPs is reduced from 35 to 5 nm with the addition of eosin, and the size of AuNPs then increased to 26 nm upon adding BSA to the mixture of ANPs and eosin. These fluorescence fluctuations may be used for trace identification and biosensing.