Sensitive, precise, and accurate HPLC method developed for levodopa analysis in methanolic extract of Mucuna utilis. Sample preparation followed by separation on Eurosphere C18 column using methanol:0.5% acetic acid (70:30 v/v). Absorbance measured at 284 nm. Method validated for linearity, precision, accuracy, LOD, and LOQ; suitable for quantitative determination of levodopa in herbal extracts. © 2017 Elsevier B.V., All rights reserved.