A water-soluble and electroactive composite - Pt nanoparticles/polyelectrolyte-functionalized ionic liquid (PFIL)/graphene sheets (GS) nanocomposite was synthesized in one pot. The structure and composition of the Pt/PFIL/GS nanocomposite were studied by means of ultraviolet-visible (UV-vis) and X-ray photoelectron spectra (XPS). Scanning electron microscopy (SEM) and transmission electron microscope (TEM) images reveal Pt nanoparticles are densely dispersed on the transparent thin PFIL-functionalized graphene sheets. The obtained Pt/PFIL/GS nanocomposite-modified electrode was fabricated to simultaneously determine ascorbic acid (AA) and dopamine (DA) by cyclic voltammetry. It is worthwhile noting that the difference between the two peak potentials of AA and DA oxidation is over 200mV, which leads to distinguishing AA from DA. The detection of increasing concentrations of AA in the presence of DA and the oxidation of continuous addition of DA in the presence of AA were also studied using differential pulse voltammetry. The proposed sensor in real sample analysis was also examined in human urine samples. Three independent oxidation peaks appear in urine sample containing AA and DA. Therefore, the Pt/PFIL/GS nanocomposite might offer a good possibility for applying it to routine analysis of AA and DA in clinical use.