In this study, a new method for extracting and isolating squalene from pumpkin seed oil was developed using magnetic nanoparticles (MNPs). The inhibition effect of extracted squalene on angiogenesis was in- vestigated in chick chorioallantoic membrane (CAM). Squalene extraction from pumpkin seed oil was per- formed in two steps: fatty acids (FAs) separation via hydrothermal adsorption on the magnetic iron oxide nanoparticles and squalene extraction using solvent extraction method at different temperatures. In the first step, the neural network modeling was applied to predict the optimum range of operating param- eters for achieving squalene with the highest purity. Results of transmission electron microscopy (TEM) analysis showed that under optimum conditions, the average diameter of the MNPs coated with FAs was about 20 nm. In the second step, the purity of squalene was estimated at 93% with a separation recovery percentage of 83.24% at the optimum temperature of 25 °C to prevent squalene oxidation. The inhibition effect of extracted squalene on angiogenesis in CAM suggested that squalene extracted from pumpkin seed oil had a significant effect on the inhibition of angiogenesis in the chicken embryo chorioallantoic membrane. Finally, this process yielded two valuable products, i.e. squalene with high-purity and MNPs coated with FAs.

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