In dietary contexts, fish are renowned for the diverse array of unsaturated fatty acids they provide to the body. While data regarding the fatty acid composition of freshwater fish are readily available in most developed countries, research on African freshwater fish remains notably limited. In this study, we investigated the fatty acid composition of fifteen freshwater fish species from Africa, primarily Afrotropical regions. Lipids were extracted using a solvent mixture of methanol, water, and chloroform. Subsequently, total fatty acids were determined by saponification, and the fatty acid profile was analyzed via GC–MS following methylation in the presence of BF3 and methanol. The results revealed lipid levels in fish fillets ranging from 1.99 % for T. guineensis to 6.41 % for M. bananensis. Total fatty acid percentages of lipids were found to be 81.40 ± 9.15 % for S. nigripinnis, 83.75 ± 6.12 % for L. niloticus, and 92.99 ± 11.30 % for C. dageti. The fatty acid profiles of 15 properly identified fish species have been determined, with the majority being identified for the first time. The composition of health-important omega-3 fatty acids EPA and total DHA ranged from 4.82 ± 0.2 % for Cyprinus lepidotus to 17.43 ± 2.50 % for Mugil bananensis. Unsupervised multivariate data analysis of fatty acid profiles of the different fish species showed significant differences. When the fish were grouped into 4 categories based on their similarities, 12 discriminant fatty acids characteristic of each group were identified. Among these fatty acids are omega-6 acids such as γ-linolenic acid and arachidonic acid, and omega-3 7,10,13,16,19-docosapentaenoic acid (DPA). This research opens avenues for consumers to make informed dietary choices aligned with their specific fatty acid needs and preferences regarding fish consumption.