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Screening for Forensically Relevant Drugs Using Data-Independent High-Resolution Mass Spectrometry

NCJ Number
Date Published
9 pages

The research presented in this article sought to evaluate an untargeted workflow for broad toxicological screening of the most recent Tier I drugs of abuse and metabolites; the article describes method performance measures, as well as the assessment of the extraction recovery, ion suppression or enhancement, and precision.


Forensic and clinical laboratories are expected to provide a rapid screening of samples for a wide range of analytes; however, the ever-changing landscape of illicit substances makes analysis complicated. There is a great need for untargeted methods that can aid these laboratories in broad-scope drug screening. Liquid chromatography hyphenated with high-resolution mass spectrometry (LC-HRMS) has become a popular technique for untargeted screening and presumptive identification of drugs of abuse due to its superior sensitivity and detection capabilities in complex matrices. An untargeted extraction and data acquisition method was evaluated for the broad screening of high-priority drugs of abuse in whole blood. A total of 35 forensically relevant target analytes were identified and extracted at biologically relevant low and high concentrations from whole blood using supported liquid extraction. Data-independent acquisition was accomplished using ultraperformance liquid chromatography and a quadrupole time-of-flight mass spectrometry. Results were acceptable for screening assays, with limits of detection at or below the recommended low-concentration cutoffs for most analytes. Analyte ionization varied from 30.1 to 267.6 percent (average: 110.5 percent) at low concentrations and from 8.6 to 383.5 percent (average: 93.6 percent) at high concentrations. Extraction recovery ranged from 8.5 to 330.5 percent (average: 105.3 percent) at low concentrations and from 9.4 to 127.5 percent (average: 82.7 percent) at high concentrations. This variability was also captured as precision, ranging from 4.7 to 135.2 percent (average: 36.5 percent) at low concentrations and from 0.9 to 59.0 percent (average: 21.7 percent) at high concentrations. The method described here is efficient and effective for qualitative forensic toxicology screening, as demonstrated by analysis of 166 authentic suspected impaired driver and postmortem specimens. That said, it is critical that laboratories establishing untargeted LC-HRMS screening assays be aware of the strengths and limitations across diverse drug categories and chemical structures. (Published Abstract Provided)

Date Published: January 1, 2024