This publication describes the development and validation of a single confirmatory assay to detect human blood, saliva, and semen using high-performance liquid chromatography and multiple-reaction monitoring mass spectrometry.
Researchers at the New York City Office of Chief Medical Examiner (OCME) have developed and validated a single confirmatory assay to detect human blood, saliva, and semen using high-performance liquid chromatography and multiple-reaction monitoring mass spectrometry. The assay uses a multi-tiered system to confirm each fluid, targeting proteins, protein fragments and peptides. There are three marker proteins from blood, three from semen, and four from saliva; detection of multiple peptides from each protein and four fragment ions from each peptide increases assay confidence. The validation was performed with diluted and degraded body fluid samples, which were used to prepare dried mock forensic stains for analysis. Non-targeted body fluids (e.g., vaginal fluid) and blood, saliva, and semen from other species were also tested. The assay can also detect and distinguish body fluid mixtures. The OCME successfully validated their body fluid assay and plans to publish their protocol in the future. This assay has been approved by the ANSI National Accreditation Board and the New York State Commission on Forensic Science and is currently being used in casework at the OCME.