A simple method has been developed for the isolation of Levodopa from the seeds of Mucuna pruriens. The isolated Levodopa was characterized by UV, TLC, FTIR, LC-MS/MS, HPLC and HPTLC. Quantification of Levodopa was done in two varieties of seeds and a single component herbal formulation containing Mucuna pruriens. The quantification was done by development of a blue colour chromogen of Levodopa using Folin – Ciocalteu Reagent (FCR) in presence of 1% sodium carbonate solution. The λmax of the blue chromogen was found at 731.4nm. The developed method was validated according to ICH guidelines. The method was found to be highly specific, simple, accurate, and reproducible.
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