Folic acid (FA) as one of the water-soluble vitamins contributes to the construction of healthy cells, as FA deficiency is one of the leading causes of anaemia. Based on reports, reduced folate level can lead to development of carcinogenesis. Hence, its analysis from the clinical point of view is highly demanded. In the present work, NiFe2O4 nanoparticles was successfully synthesized and used for modified novel voltammetric sensor for determination of folic acid. Differential pulse voltammetry response shows the linear increment of oxidation signals with an increase in the concentration of folic acid in the range of 1.0 × 10-7-5.0 × 10-4 M with limit of detection 3.4 × 10-8 M. The modified electrode displays an excellent selectivity towards the determination of FA even in the presence of various interfering species. Finally, the screen printed electrode (SPE) consists of three main parts which are a graphite counter electrode, a silver pseudo-reference electrode and a graphite working electrode modified by NiFe2O4 nanoparticles (NFO) was applied for FA determination folic acid tablet and urine samples whose accuracy was attested by means of addition and recovery assays (97.0-103.5%) as well as by differential pulse voltammetry.