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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)

Separation of Polyphenols from Jordanian Olive Oil Mill Wastewater