Impact of Cold Ischemia on the Proteome and Metabolome of Tissue Samples and Preanalytical Quality Control Using Mass Spectrometry Imaging.

Cold ischemia significantly affects tissue sample quality and analytical accuracy. However, research on the effects of cold ischemia on tissue proteomics and metabolomics is limited, and efficient tools for assessing tissue quality are lacking. In this study, mass spectrometry (MS)-based proteomics and metabolomics were employed to evaluate the impact of cold ischemia times and temperature, and MS imaging (MSI) was explored as a high-throughput platform for tissue quality assessment. Our findings indicated that the proteome exhibits greater overall stability than the metabolome and that cold ischemia at 25 °C causes tissue molecular degradation more rapidly than at 4 °C. We also identified functional pathways particularly sensitive to cold ischemia, including the upregulation of cellular detoxification and small-molecule metabolism, alongside the inhibition of nucleotide excision repair functions. Additionally, MALDI MSI can identify the change in certain highly unsaturated lipids caused by cold ischemia. Specifically, phosphatatidylcholines (40:6) and phosphatatidylcholines (34:3) showed sensitivities of 91.67% and specificities of 88.89% for identifying ″low-quality″ leiomyoma tissues, indicating their potential as quality biomarkers. These insights can guide sample collection and selection strategies based on analytical objectives and provide a theoretical basis for preanalysis quality management.