Abstract:In order to develop a rapid, sensitive and accurate real-time detection technique to assess the degree of deterioration of pork during cold chain breaks, in this study, lipidomic data of pork samples with different cold chain interruption times were analyzed by rapid evaporative ionization mass spectrometry (REIMS) combined with an intelligent knife (iKnife). The results showed that the relative abundance of the characterized ions differed significantly among the samples (P < 0.05). Temperature fluctuations occurring during cold chain logistics of chilled meat led to lipid oxidation. A total of six ions were identified as potential markers, including m/z 353.2703, m/z 507.44259 ([FAHFA(16:1(9Z)/5-O-16:0)]?), m/z 352.2861 ([N-palmitoyl proline]?), m/z 271.2284 ([2-hydroxy palmitic acid]?), m/z 331.2494 ([9,10,13-trihydroxy-Octadecanoic acid]?), and m/z 309.2438 ([9R-HODE, methyl ester]?), which could serve as markers for cold chain deterioration. By incorporating the typical marker m/z 353.2703 into the REIMS scanning scope through target enhancement, the overall recognition accuracy improved significantly, increasing from 55.77% (non-targeted acquisition mode) to 97.60% (target enhancement scanning). In conclusion, the target-enhanced iKnife-REIMS method can accurately discriminate cold chain interrupted pork according to different levels of lipid deterioration, providing near real-time results.