In this work, a simple, selective, and sensitive probe for hemoglobin based on the quenched fluorescence of silicon nanoparticles (SiNPs) was fabricated. The SiNPs were synthesized by a simple hydrothermal treatment from N-[3-(trimethoxysilyl)propyl]ethylenediamine and sodium citrate. The as-prepared SiNPs exhibited good water-solubility and high fluorescence with the quantum yield of 70%. The fluorescence of the SiNPs could be remarkably quenched by hemoglobin. A wide linear range was obtained from 50 nM to 4000 nM with a LOD of 40 nM. The quenching mechanism was investigated by UV-Vis absorption spectrometry and time-resolved fluorescence spectrometry.