The present study compared the fatty acids, cholesterol, low density lipoprotein (LDL), and high density lipoprotein (HDL) of four commercially important fresh-water fish species, Clarias gariepinus (CG), Oreochromis niloticus (ON), Synodontis budgetti (SB), and Mormyrops deliciosus MD), using Gas Chromatographic and High Performance Lipid Chromatographic methods. Results showed that the percent polyunsaturated fatty acids (PUFAs) determined from CG were, 35.709, 14.310, 45.845 and 32.345, from Amassoma, Swali, Tombia (in Bayelsa State, and Lagos rivers respectively. The prominent omega-3 fatty acid determined were eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA); while the major omega-6 fatty acid was eicosatetraenoic acid (AA). Cholesterol content of fishes from the study rivers ranged from 5.080 to 5.570 (mg/100g DW) and was more or less significantly lower than that from Lagos river (in a populous city in the South-west of Nigeria), (p < 0.05). Cholesterol, HDL and LDL content in the four fish species were according to the decreasing order CG > ON >SR .MD. For the HDL content of fish, higher value was obtained from Amassoma than those of Swali and Tombia rivers but (with the exception of that of O. niloticus in Amassoma) were all found to be significantly lower than that of Lagos, (p < 0.05). Fishes from Amassoma and Lagos rivers were prominent in n-3 PUFA particularly DHA and EPA; while those from SR showed the lowest levels. This information should find application not only for institutional feed and culture development in fisheries but for individual / institutional nutrition guide and dietary recommendation.