Recently, Costa et al. published an article about promising biomarkers for estimating the postmortem interval. Instead of postmortem blood, antemortem blood was putrefied in vitro by exposing the blood to a temperature gradient. However, in this way several other influencing factors were excluded, hence, the accuracy of the proposed model is doubtful. Therefore, the aim of this comment is to discuss the methodology, results and shortcomings of the study of Costa et al.