Automated enzymatic hydrolysis of urine samples for improved systematic toxicological analysis of drug-facilitated sexual assault cases

IF 2.6 3区 医学 Q2 BIOCHEMICAL RESEARCH METHODS
Kathrine Skov, Sys Stybe Johansen, Kristian Linnet, Marie Katrine Klose Nielsen
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Abstract

Drug-facilitated sexual assault (DFSA) is characterized by victim incapacitation due to intoxicating substances. Detection of single drug exposure from DFSA requires a systematic toxicological analysis strategy including sensitive methods covering a broad spectrum of substances. The aim of this study was to develop and validate an UHPLC–MS/MS screening method for analysis of samples from DFSA cases and incorporate an automated enzymatic pre-treatment of urine samples into a robotic sample preparation for an efficient laboratory workflow. The screening method included 144 drugs of abuse, pharmaceuticals, and metabolites relevant to DFSA. The use of a recombinant enzyme showed an efficient glucuronide hydrolysis with an average parent drug recovery of 97%. Investigation of matrix effect showed no pronounced ion enhancement or suppression for most analytes (96%), and extraction recovery was above 80% for 97% of analytes. Process efficiency ranged from 50% to 138% for most analytes. The LODs ranged from 0.0001 mg/L to 2 mg/L depending on analyte, and most analytes met the SOFT recommended minimum performance limits. The validated method was applied to authentic suspected DFSA cases (n = 38). Results showed that drugs of abuse, benzodiazepines, and antidepressants were most commonly found in suspected DFSA cases. Incorporation of an automated enzymatic hydrolysis step during sample preparation enables a fast and simple workflow for simultaneous analysis of blood and urine samples for an improved systematic toxicological analysis strategy for DFSA cases.

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对尿液样本进行自动酶水解,以改进对借助药物的性侵犯案件的系统毒理学分析。
借助药物的性侵犯(DFSA)的特点是受害者因服用麻醉药物而丧失能力。检测 DFSA 中的单一药物暴露需要系统的毒理学分析策略,包括涵盖广泛物质的灵敏方法。本研究旨在开发和验证一种超高效液相色谱-质谱/质谱筛选方法,用于分析 DFSA 案件的样本,并将尿液样本的自动酶预处理纳入机器人样本制备中,以实现高效的实验室工作流程。筛查方法包括与 DFSA 相关的 144 种滥用药物、药品和代谢物。重组酶的使用显示了高效的葡萄糖醛酸水解作用,母体药物的平均回收率为 97%。基质效应调查显示,大多数分析物(96%)没有明显的离子增强或抑制现象,97%的分析物的萃取回收率高于 80%。大多数分析物的处理效率在 50% 到 138% 之间。根据分析物的不同,最低检测限为 0.0001 毫克/升至 2 毫克/升,大多数分析物符合 SOFT 建议的最低性能限。对真实的疑似 DFSA 病例(n = 38)采用了经过验证的方法。结果表明,在疑似 DFSA 病例中最常发现的是滥用药物、苯二氮卓类药物和抗抑郁药物。在样品制备过程中加入自动酶水解步骤可实现快速简便的工作流程,同时分析血液和尿液样品,从而改进DFSA病例的系统毒理学分析策略。
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来源期刊
Drug Testing and Analysis
Drug Testing and Analysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL
CiteScore
5.90
自引率
24.10%
发文量
191
审稿时长
2.3 months
期刊介绍: As the incidence of drugs escalates in 21st century living, their detection and analysis have become increasingly important. Sport, the workplace, crime investigation, homeland security, the pharmaceutical industry and the environment are just some of the high profile arenas in which analytical testing has provided an important investigative tool for uncovering the presence of extraneous substances. In addition to the usual publishing fare of primary research articles, case reports and letters, Drug Testing and Analysis offers a unique combination of; ‘How to’ material such as ‘Tutorials’ and ‘Reviews’, Speculative pieces (‘Commentaries’ and ‘Perspectives'', providing a broader scientific and social context to the aspects of analytical testing), ‘Annual banned substance reviews’ (delivering a critical evaluation of the methods used in the characterization of established and newly outlawed compounds). Rather than focus on the application of a single technique, Drug Testing and Analysis employs a unique multidisciplinary approach to the field of controversial compound determination. Papers discussing chromatography, mass spectrometry, immunological approaches, 1D/2D gel electrophoresis, to name just a few select methods, are welcomed where their application is related to any of the six key topics listed below.
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