Fat content and fatty acid composition are important nutritional properties of shea fruits. Farmers inUganda report the presence of local shea tree ethno-varieties, but it is necessary to investigate their relative fat content and fatty acid composition to evaluate the economic importance of these ethno-varieties. Near infrared spectrophotometry (NIRS) was used to determine the fat content as well as the fatty acid composition of 44 ethno-varieties.Wet chemistry (soxtec petroleum - ether fat extraction and gas chromatography)methodswere used to validate the results fromNIRS. Fat content ranged from43.9%to 58.4%while fatty acid composition was dominated by oleic (47-62%) and stearic acid (25-38%). Other fatty acids present were palmitic, vaccenic, linoleic, linolenic and arachidic acids. There was no significant difference in stearic, palmitic and oleic acid composition between ethno-varieties. However, significant variation of fat content, vaccenic and linoleic acids was observed between some ethno-varieties, perhaps due to locality, climatic and tree-totree differences. These findings can be utilized for the selection of ethno-varieties that are suitable for commercial production of shea oil in Uganda.