Total volatile basic nitrogen (TVB-N) value is a recognized national standard for evaluating physical and chemical indicators of meat freshness. In this paper, the measurement uncertainty for the amount of TVB-N in fresh meat was evaluated using semi-micro method of Kay’s fixed nitrogen in order to accurately measure the volatile basic nitrogen content of the fresh meat and assess the freshness of meat objectively. The evaluation result indicated that the major factors of uncertainty were the actual consumption volume of hydrochloric acid for sample solution and the repeatability of measurement outcome.