The existence of carcinogenic nitrites in food and the natural environment has attracted much attention. Therefore, it is still urgent and necessary to develop nitrite sensors with higher sensitivity and selectivity and expand their applications in daily life to protect human health and environmental safety. Herein, one-dimensional honeycomb-like carbon nanofibers (HCNFs) were synthesized with electrospun technology, and their specific structure enabled controlled growth and highly dispersed bismuth nanoparticles (Bi NPs) on their surface, which endowed the obtained Bi/HCNFs with excellent electrocatalytic activity towards nitrite oxidation. By modifying Bi/HCNFs on the screen-printed electrode, the constructed Bi/HCNFs electrode (Bi/HCNFs-SPE) can be used for nitrite detection in one drop of solution, and exhibits higher sensitivity (1269.9 μA mM−1cm−2) in a wide range of 0.1~800 μM with a lower detection limit (19 nM). Impressively, the Bi/HCNFs-SPE has been successfully used for nitrite detection in food and environment samples, and the satisfactory properties and recovery indicate its feasibility for further practical applications.