This research aims at separation of polyphenols from Jordanian olive mill wastewater which have possible applications in pharmaceutical industry. The phenolic compounds were isolated using silica column chromatography based on using different solvents after extracting the acidified solution with n-hexane and ethyl acetate. The structural elucidation of the separated compounds was achieved using 1H-NMR, 13C-NMR and mass spectrometry. The concentrations of these compounds were determined by GC-MS after derivatization with N, O-bis(trimethylsilyl)trifluoroacetamide (BSTFA). The concentrations of the main isolated phenolic compounds in the Jordanian olive mill wastewater were ferulic acid (93.6？mg/L), trans-cinnamic acid (105.3？mg/L), p-coumaric acid (117.0？mg/L), vanillic acid (128.7？mg/L), caffeic acid (140.4？mg/L), tyrosol (210.6？mg/L), and hydroxytyrosol (315.9？mg/L). 1. Introduction Olive mill wastewater (OMWW) is a dark red-to-black-colored, mildly acidic liquid of high conductivity, obtained from mechanical olive processing during olive oil production [1]. Three phase-extraction systems involve the addition of large amounts of water (up to 50？L/100？kg olive paste), resulting in the worldwide production of more than 30 million m3 per year of OMWW [2]. This represents a great environmental problem, since this by-product is characterized by a high inorganic and organic load. Organic substances found in OMWW include sugars, tannins, phenolic compounds, polyalcohols, pectins, and lipids [3]. The toxicity, the antimicrobial activity, and the consequent difficult biological degradation of OMWW are mainly due to the phenolic fraction [2, 4]. The treatment of OMWW is extremely difficult due to its large volume and the high concentration of organic matter. The major factor of the environmental problems imposed by the OMWW is the high concentration of polyphenols. These compounds are difficult to decompose [5, 6] and present phytotoxicity [7, 8], toxicity against aquatic organisms [9], or suppression of soil microorganisms [10]. The olive fruit is very rich in phenolic compounds, but only 2% of the total phenolic content of the olive fruit passes in the oil phase, while the remaining amount is lost in the OMWW (approx. 53%) and in the pomace (approx. 45%) [11]. In general, polyphenols are thought to deliver health benefits by several mechanisms, including: (1) direct free radical quenching, (2) protection and regeneration of other dietary antioxidants, (3) chelation of metal ions [12]. So they act as antioxidant [13], antibiotic/antiviral [12],
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