Coffee Arabica is highly cultivated in different areas of Amhara region, specifically East Gojjam, West Gojjam, and Awi zones. However, this coffee has not been well characterized and branded for the national and international market. Therefore, this study was aimed to profile the fatty acid contents of green coffee beans from West Gojjam, Awi, and East Gojjam zones of the Amhara region. Twenty four samples of green coffee (Coffea Arabica) beans collected from the major producing zones, comprising various Woredas, were studied for variations in their fatty acid compositions. The lipid fractions of the powdered green coffee beans were extracted using solvent extraction method and the fatty acid portion of the lipid fraction was derivatized to the corresponding fatty acid methyl esters (FAMEs). The individual fatty acids were identified and quantified using gas chromatography coupled with mass spectrometry (GC-MS) following standard procedures. Data were analyzed with SPSS version 22.0. Twenty three fatty acid types were detected: Among these myristic (C14:0), pentadecanoic acid (C15:0), palmitic (C16:0), margaric acid (C17:0), stearic (C18:0), oleic (C18:1), linoleic (C18:2), arachidic (C20:0), gondoic acid (C20:1),14-methyleicosanoic acid (C21:0), behenic acid (C22:0), tricosanoic acid (C23:0) and lignoceric acid (C24:0) were the major ones. There was a significant difference (p<0.05) between zones and woredas in the concentration of total fatty acids. The total fatty acid contents of West Gojjam (102. 74 mg g -1 ) significantly higher than from East Gojjam and Awi zones were 84.56 and 90.55 mg g -1 respectively and Bure city administration coffees samples from West Gojjam zone contained (123.54 mg g -1 ) of the total fatty acid content significantly higher than the rest woredas coffee types ranged from (78.45−104.88 mg g -1 ). Myristric, palmitic, linoleic, stearic, arachidic and trieicosanoic acid were identified as the most important compounds for discrimination of coffee samples based on their geographical origin. The studied fatty acids provide adequate information for use as descriptors of the geographical origin of coffee beans from Amhara region. Keywords: Green Coffee Beans; Fatty acids; Geographical origin.