Vitamin B6 (Vit B6), also known as pyridoxine, is pivotal in fundamental physiological and metabolic processes within the body. Insufficient levels of this essential nutrient may contribute to various health complications. We introduce an electrochemical sensor designed to determine Vit B6 levels precisely. This sensor is constructed through a two-step process: first, by modifying a bare carbon fiber paper electrode (CFP) with poly(2-aminothiazole) (PAT), and second, by electrodepositing platinum nanoparticles onto the modified electrode surface, giving the final working electrode- Pt/PAT/CFP. Electrochemical impedance spectroscopy (EIS) and Cyclic voltammetry (CV) were utilized to examine the electrochemical characteristics of the developed sensor. The characterization of the sensor was done through a range of analytical techniques, including X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), and optical profilometric studies. Furthermore, we optimized the sensor’s performance by assessing the impact of pH, scan rates, and analyte concentrations. The sensor showed a wide linear dynamic range of 5.0 nM–80 µM and a low detection limit of 0.054 µM. We have successfully quantified Vit B6 levels in tablet formulations and dried palm date fruits. The outcomes of this study hold the promise of substantial progress in Vit B6 quantification, with far-reaching implications across pharmaceuticals, healthcare, and nutritional science.