Biosensors have emerged as powerful tools in ensuring the safety and quality of food products, addressing critical issues such as contamination, adulteration, and spoilage. This chapter provides a comprehensive overview of the application of non-destructive biosensors in food safety and quality testing, exploring their underlying principles, recent advancements, challenges, and future prospects. The basis of biosensors is their capacity to selectively identify target analytes of interest by using biological recognition components, such as enzymes, antibodies, or nucleic acids. Biosensors, when coupled with suitable transduction mechanisms, convert these biochemical interactions into quantifiable signals, providing prompt, sensitive, and on-site detection capabilities that are essential for preserving public health and consumer trust in the food supply chain. Put it all up, biosensors are essential instruments for guaranteeing the security, authenticity, and excellence of food items, tackling changing obstacles in a dynamic and intricate worldwide food distribution network. Biosensors provide quick, accurate, and targeted detection capabilities that are essential for safeguarding consumer confidence and public health. They achieve this by utilizing novel transduction mechanisms and biological recognition features. Realizing the full potential of biosensors in food safety and quality testing will require cooperation between academics, industry, and regulatory bodies as we continue to push the boundaries of biosensor technology.