A sensitive electrochemical analysis of riboflavin (RFN) was done in 0.1 M phosphate buffer solution (PTBS) by constructing a modest and green electrochemically polymerised (EP) valine (VLN) modified graphite paste electrode (GPE). The electrochemical, morphological, resistive, and conductive characteristics of modified electrode (EP(VLN)MGPE) and bare graphite paste electrode (BGPE) surfaces were clarified through cyclic voltammetry (CV), field emission scanning electron microscopy (FE-SEM), and Electrochemical impedance (EI) methods. The EP(VLN) initiated electrode surface parades excellent sensitivity, steadiness, and reproducibility for the redox action of RFN with enhanced peak currents as associated to BGPE. The oxidation peak currents of RFN are improved with increased concentrations of RFN in the range from 2.0 µM to 40.0 µM and the attained limit of detection (LOD) and limit of quantification (LOQ) are found to be 28.69 × 10-8 M and 95.66 × 10-8 M, respectively. The prepared EP(VLN)MGPE was beneficial for the quantitative and qualitative investigation of RFN in medicinal sample.