Vitamin D deficiency is becoming a severe burden worldwide, the rapid lifestyle changes are an important cause of that. As a result, rapid, cost-effective, and discriminating sensors for the determination of vitamin D are highly required. Herein, we report, a non-enzymatic sensitive electrochemical method for the determination of vitamin D3 using boron-doped microporous carbon (BMC) as a sensing matrix. The Boron was systematically integrated into the microporous carbon (MC) matrix to increase the electrochemical property. Among the synthesized materials, the MC modified with 5 wt.% boron showed better sensor performance than the others. The kinetics and concentration studies indicate that the rate of electro oxidation on the surface of the modified electrode is significantly faster than that of the unmodified electrode. The scan rate effect and pH studies provide the number of electron transfers, transfer coefficient, and diffusion coefficient of vitamin D3 which are 2, 0.52, and 18.49x10–12 cm2s-1, respectively. The electrolytic pH had a large impact on the oxidation potential, and its activity was significantly closer to that of the previously reported boron-doped diamond electrode. The BMC-5 modified electrode has a great linear range (0.5 μM–42 μM) and a low detection limit (1.45 μM). Hence, from this study, it is concluded that the low-cost and stable BMC is an alternate matrix for the determination of vitamin D3.