The excessive input of heavy metals such as vanadium (V) into the environment has been one of the consequences of global industrial development. Excessive exposure to V can pose a potential threat to ecological safety and human health. Due to the heterogeneous composition and reactivity of the various elements in soils and sediments, quantitative analysis of the chemical speciation of V in different environmental samples is very complicated. The analysis of V chemical speciation can further reveal the bioavailability of V and accurately quantify its ecotoxicity. This is essential for assessing for exposure and for controlling ecological risks of V. Although the current investigation technologies for the chemical speciation of V have grown rapidly, the lack of comprehensive comparisons and systematic analyses of these types of technologies impedes a more comprehensive understanding of ecosystem safety and human health risks. In this review, we studied the chemical and physical extraction methods for V from multiple perspectives, such as technological, principle-based, and efficiency-based, and their application to the evaluation of V bioavailability. By sorting out the advantages and disadvantages of the current technologies, the future demand for the in situ detection of trace heavy metals such as V can be met and the accuracy of heavy metal bioavailability prediction can be improved, which will be conducive to development in the fields of environmental protection policy and risk management.