The separation and estimation of natural products by chromatographic techniques such as high-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC) are widely preferred. The present work reveals the quantification of morphine (a regulated narcotic agent) in the Ayurvedic formulation ̀Kamini Vidrawan Raś using reversed-phase HPLC (RP-HPLC). The results obtained herein were compared with the results of earlier reported work using HPTLC. The HPTLC separation of morphine was performed on an aluminum-backed layer of silica gel 60 F254 using ethyl acetate–methanol–ammonia solution as the mobile phase, while RP-HPLC was performed on Kromasil C8-column (150 mm × 4.6 mm, 5 µm) using a mobile phase comprising of N-heptane sulfonate sodium salt–acetonitrile (70:30, v/v) at a flow rate of 1.5 mL min-1. The International Conference on Harmonisation (ICH) guidelines were followed for validation of both the chromatographic methods. Both the developed chromatographic methods are simple, rapid, accurate, and sensitive.