The study combines the reaction of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) with reduce discoloration and high-performance thin layer chromatography to establish a new method for the determination of rutin in healthcare products by HPTLC-DPPH. The effects of DPPH color reaction time, mobile phase, and optical density scanning parameters on chromatographic separation and densitometric results are investigated as well as recovery and precision of the method. The standard addition recovery rate of the method is 82%~97%, and the precision RSD is <10%. It was demonstrated that the method is able to circumvent the interferences from sample matrices with a simplified sample pretreatment. Therefore, the method is suitable for rapid detection of rutin in healthcare products.