We report that ultra-small, monodisperse, water-dispersible magnetite (Fe3O4) nanoparticles can be synthesized by a facile one-pot approach using trisodium citrate as crystal grain growth inhibitor and stabilizer in polyol solution. The resultant Fe3O4 nanoparticles exhibit an excellent long-term colloidal stability in various buffer solutions without any modification. They are also superparamagnetic at room temperature and their magnetic property relies heavily on their size. Due to the low magnetization and good water-dispersibility, the 1.9 nm-sized Fe 3O4 nanoparticles reveal a low r2/r1 ratio of 2.03 (r1 = 1.415 mM-1 s-1, r 2 = 2.87 mM-1 s-1), demonstrating that they can be efficient T1 contrast agents. On the other hand, because of the excellent magnetic responsivity, the 13.8 nm-sized Fe3O4 nanoparticles can be readily modified with nitrilotriacetic acid and used to separate the protein simply with the assistance of a magnet. In addition, these Fe3O4 nanoparticles may be useful in other fields, such as hyperthermia treatment of cancer and targeted drug delivery based on their size-dependent magnetic property and excellent stability. ? 2013 The Royal Society of Chemistry.