In this work, we synthesized NixCo1-xFe2O4 (0 ≤ x ≤ 0.8) magnetic nanoparticles (MNPs) by using ultrasonication-assisted reverse chemical co-precipitation method. The X-ray diffraction (XRD) patterns confirm the single-phase with mixed spinel structure of the NixCo1-xFe2O4 (0 ≤ x ≤ 0.8) MNPs with the crystallite size range between 12 nm - 17 nm. The molecular dynamics and oleic acid coated Ni-substituted CoFe2O4 magnetic nanoparticles were confirmed by FTIR measurements. The surface composition of NixCo1-xFe2O4 (0 ≤ x ≤ 0.8) MNPs (Ni, Co, Fe and O) and their oxidation states were estimated using X-ray photoelectron spectroscopy. The NixCo1-xFe2O4 (0 ≤ x ≤ 0.8) MNPs are spherical in shape and polycrystalline in nature, confirmed by FESEM and HRTEM measurements. The Ni-substituted CoFe2O4 MNPs with higher zeta potential (ζ) from −24 mV to -41 mV with an increase in Ni concentration, signifies the stable colloidal stability due to the electrostatic repulsion between MNPs. The DC magnetization (M-vs-H) measurements reveal the ferrimagnetic behavior of NixCo1-xFe2O4 (0 ≤ x ≤ 0.8) MNPs, which suppress the saturation magnetization (MS) from 48.6 emu/g to 5.3 emu/g with the enhanced coercivity (HC) from 335 Oe to 1700 Oe with the substitution of Ni2+ ions is in corroboration with electron paramagnetic resonance (EPR) measurements. The enhanced EPR resonance field (Hres = 2335 Oe - 3261 Oe) with the substitution of Ni2+ ions is attributed to the generation of oxygen vacancies and defects, which are predominant in Ni0.8Co0.2Fe2O4, resulting in significantly enhanced electrochemical performance. Therefore, the three-electrode system was utilized to investigate the electrochemical properties of NixCo1-xFe2O4 (0 ≤ x ≤ 0.8) MNPs. The Ni-substituted CoFe2O4 MNPs have demonstrated significantly enhanced capacity retention of 77 % even after 5000 cycles in 1 M H2SO4 electrolyte solution. NixCo1-xFe2O4 (0 ≤ x ≤ 0.8) MNPs exhibit an excellent specific capacity (Cs) of 794.5 F/g at a current density of 1 A/g, signifies the rate of redox reaction at surface electrolyte interface (SEI) is greatly influenced by the surface to volume ratio. Thus, with the enhancement of Ni2+ ions, redox reaction kinetics might become more efficient resulting in the enhanced charge storage capacity.