Abstract: | Vitamin D play a vital role in health, therefore, it is necessary to find a sensitive, selective as well as quick and easy technique for its determination. This review focuses on the Vitamin D investigations of electrochemical biosensors that have been conducted in recent years. According to the research, the practical use of electrochemical biosensors is attributed to the existence of UV radiation and transactivation of pharmaceutical items, food, or even human blood plasma in the detection of Vitamin D from diverse samples, including Vitamin D production in nature. Among the most commonly used electrochemical biosensors for vitamin D detection are Ab-25OHD/SPE/ FMTAD, CYP27B1/GCE, SiO2/GO/Ni(OH)2/GCE, BSA/Ab-VD2/CD-CH/ITO, BSA/Anti VD/Fe3O4 PANnFs/ITO, BSA/Ab-VD/Asp-Gd2O3NRs/ITO, 25OHD Antibody, 25OHD, 25OHD Antibody, IoT Enabled Enzyme Embossed Biosensor, Au-Pt NPs/APTES/FTO and GCN- β -CD/Au nanocomposite. The proposed electrochemical biosensors utilized in the previous publications studied were based on glassy carbon, carbon dots, or carbon paste, functionalized with the various electrochemical biosensors. Further research should be conducted on existing problems and future opportunities of the present electrochemical sensors for the determination of vitamin D.