Most of the studies focused on V2O5 have been devoted to obtaining specific morphology and microstructure for its intended applications. Two dimensional (2D) V2O5 has the most valuable structure because of its unique planar configuration that can offer more active sites. In this study, a bottom-up and low-cost method that is hydrothermal combined with spin-coating and subsequent annealing was developed to prepare 2D V2O5 nanosheets film on quartz substrate. First, VOOH nanosheets were prepared by the hydrothermal method using V2O5 powders and EG as raw materials. Further, V2O5 nanosheets with an average lateral size over 500 nm and thickness less than 10 nm can be prepared from the parent VOOH nanosheets by annealing at 350 °C for 15 min in air. The prepared V2O5 nanosheets film was assembled of multiple nanosheets. The structural, morphological, microstructural and optical properties of the films were respective investigated by XRD, SEM, TEM and UV-Vis. The photodetector based on V2O5 nanosheets film shows good photoresponse with a response time of 2.4 s and a recovery time of 4.7 s.