Melia azedarach L. is a plant with wide use in folk medicine since it contains many bioactive compounds of interest. The present study aimed to extract bioactive compounds from M. azedarach fruits by a sequential process in fixed bed using various solvent mixtures. Extractions were performed at 50 degrees C and 300 bar in four sequential steps using supercritical CO2 (scCO(2)), scCO(2)/ethanol, pure ethanol, and ethanol/water mixture as solvents, respectively. The efficacy of the extraction process was evaluated by extraction yield and kinetics, and analysis of extracts by: (1) thin layer chromatography (TLC), (2) phenolics content, (3) reduction of surface tension of water, (4) gas chromatography (GC-MS), (5) electrospray ionization mass spectrometry (ESI-MS) and (6) antiviral activity. The overall extraction yield reached 45% and TLC analysis showed extracts with different composition. extract obtained from CO2/ethanol mixture (SCEE) exhibited the greatest ability to reduce surface tension of water from 72.4 mN m(-1) [1] of pure water to 26.9 mN m(-1) of an aqueous solution of 40 g L-1. The highest phenolics contents were observed in both the hydroalcoholic extract and scCO(2)/ethanolic extract. Volatile oils were not detected in the supercritical extracts by GC-MS. MS analyses identified the fatty acids: linoleic, palmitic and myristic acid in the supercritical extract (SCE), and the phenolics: caffeic acid and malic acid in the other extracts. In addition, SCE and SCEE extracts showed significant inhibition percentage against Herpes Simplex Virus Type 1. The extraction process proposed in the present study produced extracts with significant potential for application in food and pharmaceutical industries. (C) 2014 Elsevier B.V. All rights reserved.