Common carp viscera, obtained from Tikves Lake in Macedonia, was investigated as a possible source of polyunsaturated (PUFA) fatty acids. Supercritical fluid CO2 extraction (SFE-CO2) was employed for extraction of investigated bioactive components. The GC-FID analysis on the total extract obtained by supercritical fluid CO2 extraction confirmed the assumption of presence of these bioactive components. A three layer artificial neural network was created for prediction and modelling of the extraction yield of polyunsaturated fatty acids from lyophilized viscera matrixes. Operating values of pressure, temperature, mass flow of CO2 and extraction time were defined as input vectors to the ANN where PUFA extraction yield was considered as an output vector. Created ANN model provided adequate fitting of experimental data, with a correlation coefficient of 0.9968 for the entire data set. RSM-3D method was employed for mathematical modelling of the ANN output values as a function of operating variables and their interactions. (C) 2014 Elsevier B.V. All rights reserved.