Mucuna pruriens Linn. (Leguminosae), commonly known as the cowhage or velvet bean or atmagupta in India, is a climbing legume endemic in India and in other parts of the tropics including Central and South America. It is used for the management of male infertility, nervous disorders and also as an aphrodisiac. L-Dopa is commonly used as a marker compound. Literature survey reveals that there are very few validated HPTLC methods for the estimation of, L-Dopa in seeds collected from different geographical sources as well as their quantification in selected over the counter (OTC) market formulations. Hence an attempt has been made in the present study to develop, validate and apply HPTLC method for evaluation of these marker compound in seeds collected from different geographical sources along with their pharmacognostical evaluation and marketed single and polyherbal formulations.