Many studies have been conducted regarding the separation behavior of carriers (electrons and holes) because of involving the generation of superoxide radicals (O2−), hydroxyl radicals (HO), and hydrogen peroxide (H2O2) in the photocatalytic process of heteropolyacids. Instead, relatively little attention has been focused on the potential Coulomb interactions between photogenerated electrons (e−) and holes (h+). Herein, choline-phosphotungstic acid (Ch3-HPW) was synthesized via one-step acid-base neutralization reaction method, and characterized. The electronic excited state analysis of Ch3-HPW showed that the formation of singlet oxygen (1O2) was related to the electron-hole interactions in the photocatalytic process of ground state molecular oxygen (3O2) activation. Subsequently, a facile fuel photocatalytic oxidative desulfurization and extraction system was established on the basis of Ch3-HPW, air, and acetonitrile (MeCN), to better understand the 3O2 activation in specific applications. The main photocatalytic reaction conditions affecting the desulfurization process, including the amount of Ch3-HPW, the volume ratio of MeCN to model oil, the initial S-concentration, air/N2 bubbling, sulfur compounds, and fuel composition, were systematically investigated under UV radiation. The sulfur removal for model oil and straight-run gasoline in the system were 99.6% and 89.9%, respectively. The results of radical scavenger experiments, electron spin-resonance (ESR) spectroscopy, and density functional theory (DFT) calculations further demonstrated that 1O2, H2O2, and h+ played important roles in the oxidation of sulfur-containing compounds. A new method was developed for the desulfurization of liquid fuels using green and inexpensive O2 in this work to promote the development of photocatalytic process of exciton-involved HPA-based photocatalysts.