Master Program in Medical Biotechnology, Department of Laboratory Medicine and Biotechnology, Tzu Chi University, Hualien, Taiwan.
Department of Laboratory Medicine and Biotechnology, Tzu Chi University, Hualien, Taiwan.
J Anal Toxicol. 2019 Mar 1;43(2):96-103. doi: 10.1093/jat/bky071.
Benzodiazepines is a large drug group used extensively to treat sleep disorders and anxiety; these drugs are frequently associated with various crimes such as murder and drug-facilitated sexual assault. With growing use and misuse of these compounds, confirmatory assays are increasingly required in clinical laboratories. In this study, 4 β-glucuronidase enzymes were systematically evaluated for their hydrolysis efficiencies. Additionally, the matrix effects and extraction recoveries of three protein precipitation plates were systematically evaluated. The recombinant IMCSzyme β-glucuronidase enzyme showed higher hydrolysis efficiency than did β-glucuronidase from abalone, Patella vulgata and Helix pomatia. Lower ion suppression and more favorable overall recovery were observed in the Supelco protein precipitation plate than in the two other plates. Analytes were separated on a superficially porous particle column (ultra biphenyl) within 4.5 min and characterized through electrospray ionization-tandem mass spectrometry in the positive ion multiple-reaction monitoring mode. The accuracy, carryover, extraction recovery, detection limit, matrix effect, quantification limit and precision of the method were validated. Sixty opioid-positive urine samples were analyzed; a high incidence of benzodiazepines was detected in these samples. The proposed technique is robust and suitable for efficiently monitoring the numerous benzodiazepines that occur in urine samples.
苯二氮䓬类药物是一组广泛用于治疗睡眠障碍和焦虑症的药物；这些药物常与各种犯罪行为相关，如谋杀和药物辅助性性侵犯。随着这些化合物的使用和滥用不断增加，临床实验室越来越需要确证检测。在本研究中，系统评估了 4 种β-葡萄糖醛酸酶对其水解效率。此外，还系统评估了三种蛋白质沉淀板的基质效应和提取回收率。重组 IMCSzymeβ-葡萄糖醛酸酶的水解效率高于鲍鱼、鲍螺和海兔的β-葡萄糖醛酸酶。与另外两种板相比，Supelco 蛋白质沉淀板的离子抑制作用更低，整体回收率更好。分析物在 4.5 分钟内在表面多孔颗粒柱（超双苯）上分离，并通过电喷雾电离串联质谱在正离子多重反应监测模式下进行特征描述。对该方法的准确性、携带污染、提取回收率、检测限、基质效应、定量限和精密度进行了验证。对 60 份阿片类阳性尿液样本进行了分析；这些样本中检测到苯二氮䓬类药物的发生率很高。该技术具有稳健性，适用于有效地监测尿液样本中存在的大量苯二氮䓬类药物。
