This study investigates the neuropharmacological efficacy of L-DOPA isolated from Mucuna pruriens seeds, focusing on its antidepressant and antiparkinsonian activities in rodent models, to validate its traditional use in neurological disorders. Seeds underwent rigorous pharmacognostic standardization, confirming authenticity via macroscopic (mean seed dimensions: 12.34 × 9.87 × 7.65 mm) and microscopic analyses (lignified macrosclereids, no starch grains). Acidified ethanolic extraction (1:1 ethanol-water, 0.1% citric acid) yielded a 12.5% w/w crude extract (MPE), with bioactivity-guided fractionation isolating L-DOPA (12.35% w/w in ethyl acetate fraction) via column chromatography and confirmed by FTIR, HR-ESI-MS, and NMR spectroscopy. Acute oral toxicity (OECD 423) in mice established safety (LD₅₀>2000 mg/kg). Antidepressant effects were assessed in mice using Forced Swim (FST) and Tail Suspension Tests (TST), where L-DOPA (20, 40 mg/kg, p.o.) dose-dependently reduced immobility time, rivaling imipramine (15 mg/kg, i.p.). Antiparkinsonian activity was evaluated in mice via haloperidol-induced catalepsy and chronic rotenone models. L-DOPA (40 mg/kg) significantly reversed catalepsy, restored locomotor activity, and in the rotenone model, ameliorated motor deficits, elevated striatal dopamine (6.52 ± 0.27 ng/mg), and mitigated oxidative stress (reduced MDA, increased GSH/SOD). These findings confirm L-DOPA as the primary bioactive mediating Mucuna pruriens’ neuroprotective effects, supporting its potential as a natural therapeutic for depression and Parkinson’s disease, warranting clinical translation.