Forensic proteomics - a tool for the assessment of biological crime scene traces.

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Mass spectrometry (MS)-based analysis is a promising alternative for identifying biological traces in forensic investigations. In contrast to established forensic tests, MS-based analysis offers high sensitivity and specificity, and multiplexing combines several analytical approaches. We developed a workflow for identifying body fluids such as blood, saliva, semen and vaginal fluid from criminal traces in a single MS-method. Various sample preparation strategies were tested, including in-gel, in-solution, and filter-aided sample preparation-assisted digestion. To identify specific marker peptides, data-independent acquisition was performed, followed by the selection of suitable body fluid-specific peptides. This enabled the establishment of a targeted LC-MS method, incorporating seven markers for blood, six for saliva, six for semen, and seven for vaginal fluid. In parallel, classification criteria for body fluid assignment were defined. The applicability of the MS-method and classification criteria was validated in a blinded study, resulting in a 100% identification rate. The established method is applied to suitable case samples in addition to the procedures established at the State Office of Criminal Investigation of North Rhine-Westphalia, and the results of the analyses provide added value for the classification of the case samples. In addition, new markers are currently under investigation for the classification of body fluids based on blood groups and sex.

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