As an important chiral molecule for the preparation of levofloxacin, the optical purity of L-aminopropanol has a crucial effect on the pharmacology and pharmacodynamics of levofloxacin. Therefore, it is of great significance to discriminate D-aminopropanol and L-aminopropanol. In this paper, an effective aminopropanol enantiomer recognition method was established on the basis of the chiral fluorescent silicon nanoparticles (SiNPs) probe. The chiral fluorescent SiNPs were fabricated via a one-step aqueous solution synthesis strategy, which avoided multiple steps, pressurizing operation, and time-consuming postmodified procedures. Significantly, D-aminopropanol could significantly enhance the fluorescence of the chiral SiNPs, while L-aminopropanol could not affect the fluorescence of the chiral SiNPs. This could have occurred because of the stronger interaction between the chiral SiNPs and D-aminopropanol than that of L-aminopropanol. Thus, the rapid and selective recognition of the aminopropanol enantiomer was ideally realized. The mechanism of the chiral SiNPs recognizing aminopropanol was simulated by density functional theory quantum mechanical calculations. Interestingly, this was also proved by the separation of aminopropanol enantiomer using this chiral SiNPs-modified silica column in normal phase liquid chromatography. To the best of our knowledge, this is the first time that the chiral fluorescent SiNPs were synthesized and used to detect the aminopropanol enantiomer successfully. This work will inspire additional syntheses of chiral silicon nanomaterials and other nanomaterials with excellent properties and will enable application of chiral nanomaterials to other fields.