Vanadium–oxygen species are very harmful to the environment and human health, and also some vanadium–oxygen species have good biological and biomedical effects to human beings. Thus, the study of vanadium–oxygen species is very significant. The types of V–O species are very complex. So far, however, the various structures of V–O species cannot be identified clearly. In this work, the surface‐enhanced Raman spectroscopy (SERS) was used to study vanadium–oxygen species in aqueous solution directly. The SERS spectra with high enhancement factor (EF) and detail information were obtained by the enhancement substrates such as gold nanoparticles (AuNPs), Ag mirror, the Au‐deposited Pt electrodes, and rough Au electrodes. The structures of vanadium‐oxygen species in solutions with different concentration and pH had been well‐characterized firstly by the combination of SERS experiments with density functional theory (DFT) calculations. This work will promote the basic study of the V–O species, and it is also a promotion for the SERS and analytical chemistry.