Determining asphyxia as the cause of death is crucial but is based on an exclusive strategy because it lacks sensitive and specific morphological characteristics in forensic practice. In some cases where the deceased has underlying heart disease, differentiation between asphyxia and sudden cardiac death (SCD) as the primary cause of death can be challenging. Herein, Raman spectroscopy was employed to detect pulmonary biochemical differences to discriminate asphyxia from SCD in rat models. Thirty-two rats were used to build asphyxia and SCD models, with lung samples collected immediately or 24 h after death. Twenty Raman spectra were collected for each lung sample, and 640 spectra were obtained for further data preprocessing and analysis. The results showed that different biochemical alterations existed in the lung tissues of the rats that died from asphyxia and SCD and could be used to distinguish between the two causes of death. Moreover, we screened and used 8 of the 11 main differential spectral features that maintained their significant differences at 24 h after death to successfully determine the cause of death, even with decomposition and autolysis. Eventually, seven prevalent machine learning classification algorithms were employed to establish classification models, among which the support vector machine exhibited the best performance, with an area under the curve value of 0.9851 in external validation. This study shows the promise of Raman spectroscopy combined with machine learning algorithms to investigate differential biochemical alterations originating from different deaths to aid determining the cause of death in forensic practice.
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