Bates Maia N, Helm Abby E, Barkholtz Heather M
Department of Chemistry, College of Letters and Science, University of Wisconsin-Madison, 1101 University Avenue, Madison, Wisconsin 53706, United States.
Forensic Toxicology Section, Environmental Health Division, Wisconsin State Laboratory of Hygiene, 2601 Agriculture Drive, Madison, Wisconsin 53718, United States.
Chem Res Toxicol. 2024 Apr 15;37(4):571-579. doi: 10.1021/acs.chemrestox.3c00379. Epub 2024 Apr 4.
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 (10× low) 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% (average: 110.5%) at low concentrations and from 8.6 to 383.5% (average: 93.6%) at high concentrations. Extraction recovery ranged from 8.5 to 330.5% (average: 105.3%) at low concentrations and from 9.4 to 127.5% (average: 82.7%) at high concentrations. This variability was also captured as precision, ranging from 4.7 to 135.2% (average: 36.5%) at low concentrations and from 0.9 to 59.0% (average: 21.7%) at high concentrations. The method described in this work 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.
法医和临床实验室需要对各种分析物进行样本快速筛查；然而，非法药物的情况不断变化，使得分析变得复杂。迫切需要非靶向方法来帮助这些实验室进行广泛的药物筛查。液相色谱与高分辨率质谱联用（LC-HRMS）因其在复杂基质中具有卓越的灵敏度和检测能力，已成为一种用于非靶向筛查和推定鉴定滥用药物的常用技术。评估了一种非靶向提取和数据采集方法，用于全血中高优先级滥用药物的广泛筛查。使用支持液液萃取法，从全血中在生物学相关的低浓度和高浓度（低浓度的10倍）下共鉴定并提取了35种法医相关目标分析物。使用超高效液相色谱和四极杆飞行时间质谱完成数据非依赖型采集。筛查分析的结果是可接受的，大多数分析物的检测限等于或低于推荐的低浓度临界值。低浓度下分析物的离子化率在30.1%至267.6%之间（平均：110.5%），高浓度下在8.6%至383.5%之间（平均：93.6%）。低浓度下的提取回收率在8.5%至330.5%之间（平均：105.3%），高浓度下在9.4%至127.5%之间（平均：82.7%）。这种变异性也以精密度表示，低浓度下在4.7%至135.2%之间（平均：36.5%），高浓度下在0.9%至59.0%之间（平均：21.7%）。如对166份真实的疑似酒驾司机和尸检样本的分析所示，本研究中描述的方法对于定性法医毒理学筛查是高效且有效的。也就是说，建立非靶向LC-HRMS筛查分析的实验室必须了解不同药物类别和化学结构的优势和局限性，这一点至关重要。
