Effective use of quadrupole time of flight mass spectrometry in forensic toxicological testing regimes

High-resolution mass spectrometry (HRMS) has emerged as a transformative technology in forensic toxicology, enabling comprehensive drug screening through sensitivity and adaptability. Despite its potential, the lack of standardized protocols for compound identification, validation and data management limits its integration into routine workflows, particularly as laboratories strive to meet evolving regulatory standards. This study aims to address these challenges by developing, validating and implementing a rapid and comprehensive quadrupole time-of-flight mass spectrometry (QTOF-MS) method using sequential window acquisition of all theoretical fragment ion spectra (SWATH) for routine workflows in the detection and direct reporting of over 900 compounds in blood and urine samples. The context of this dissertation will include the following publications: (1) The use of sequential window acquisition of all theoretical fragment ion spectra (SWATH), a data-independent acquisition high-resolution mass spectrometry approach, in forensic toxicological regimes: A review; (2) A Validated Screening and Confirmation Method for 946 Drugs and Metabolites Using LC-QTOF-MS with SWATH Acquisition; (3) Characterization, Optimization and Selection of Identification Criteria for LC-QTOF-MS; (4) Method Design and Automation of a Forensic Toxicology LC-QTOF-MS Technique to Meet High-Throughput Routine Applications; and (5) Workflow and Application of LC-QTOF-MS in Routine Forensic Casework. Together, these efforts establish a comprehensive framework for the integration of LC-QTOF-MS into routine forensic toxicology. The outcomes not only demonstrate the method’s analytical rigor and legal defensibility but also provide a foundation for future advancements in high-resolution mass spectrometry applications.