The green coffee samples were collected from three zones in the Amhara region, assigned to ensure representation of the producing areas, namely the Awi zone, South Gondar zone, and Zegie peninsulae. A total of forty-four samples were collected. Of these, thirty samples from the Awi zone, nine samples from the Bahir Dar Administrative zone and five samples from the South Gondar Zone were collected during the 2021 crop season using a judgmental sampling technique. The Folch method was used to extract lipids from green coffee beans. 0.500 g of coffee powder was extracted with chloroform and methanol (2:1 ratio v/v) by shaking at 270 rpm for 24 hours on a platform shaker with the aid of a test tube. A total of twelve fatty acids, eight saturated and four unsaturated fatty acids were detected in all green coffee bean samples. The highest fatty acid content of green coffee beans in three zones were myristic, pentadecanoic ,palmitic, oleic ,linoleic, arachidic, gondoic, behenic, tricosanoic, and lignoceric acid. Linoleic acid was the most abundant fatty acid in the green coffee beans and accounted for 44.27– 47.31% of the total fatty acid content. The level of palmitic acid found in this study ranged from 42.87% to 53.47%.