School of Pharmacy, University of Oslo, P.O. Box 1068, Blindern, 0316, Oslo, Norway.
Department of Forensic Sciences, Oslo University Hospital, P.O. Box 4950, Nydalen, 0424, Oslo, Norway.
Anal Bioanal Chem. 2018 Aug;410(20):4967-4978. doi: 10.1007/s00216-018-1147-y. Epub 2018 Jun 8.
Benzodiazepines (BZD) and Z-hypnotics are frequently analyzed in forensic laboratories, and in 2012, the designer benzodiazepines (DBZD) emerged on the illegal drug scene. DBZD represent a particular challenge demanding new analytical methods. In this work, parallel artificial liquid membrane extraction (PALME) is used for sample preparation of DBZD, BZD, and Z-hypnotics in whole blood prior to UHPLC-MS/MS analysis. PALME of BZD, DBZD, and Z-hypnotics was performed from whole blood samples, and the analytes were extracted across a supported liquid membrane (SLM) and into an acceptor solution of dimethyl sulfoxide and 200 mM formic acid (75:25, v/v). The method was validated according to EMA guidelines. The method was linear throughout the calibration range (R > 0.99). Intra- and inter-day accuracy and precision, as well as matrix effects, were within the guideline limit of ± 15%. LOD and LLOQ ranged from 0.10 to 5.0 ng mL and 3.2 to 160 ng mL, respectively. Extraction recoveries were reproducible and above 52%. The method was specific, and the analytes were stable in the PALME extracts for 4 and 10 days at 10 and - 20 °C. No carry-over was observed within the calibration range. PALME and UHPLC-MS/MS for the determination of DBZD, BZD, and Z-hypnotics in whole blood are a green and low-cost alternative that provides high sample throughput (96-well format), extensive sample clean-up, good sensitivity, and high reproducibility. The presented method is also the first method incorporating analysis of DBZD, BZD, and Z-hypnotics in whole blood in one efficient analysis. Graphical abstract.
苯二氮䓬类药物（BZD）和 Z 类催眠药经常在法医实验室中进行分析，2012 年，出现了苯二氮䓬类药物（DBZD）这一非法药物。DBZD 是一个特殊的挑战，需要新的分析方法。在这项工作中，平行人工液膜萃取（PALME）被用于在 UHPLC-MS/MS 分析之前对全血中的 DBZD、BZD 和 Z 类催眠药进行样品制备。从全血样本中进行 BZD、DBZD 和 Z 类催眠药的 PALME 萃取，将分析物穿过支撑液膜（SLM）并进入二甲基亚砜和 200mM 甲酸（75:25，v/v）的接受溶液中。该方法是根据 EMA 指南进行验证的。该方法在整个校准范围内呈线性（R>0.99）。日内和日间准确度和精密度以及基质效应均在±15%的指南限值内。LOD 和 LLOQ 范围分别为 0.10 至 5.0ng/mL 和 3.2 至 160ng/mL。萃取回收率可重现，高于 52%。该方法具有特异性，分析物在 PALME 提取物中 4 天和 10 天在 10°C 和-20°C 下稳定。在校准范围内未观察到拖尾现象。PALME 和 UHPLC-MS/MS 用于全血中 DBZD、BZD 和 Z 类催眠药的测定是一种绿色且低成本的替代方法，可提供高通量（96 孔格式）、广泛的样品净化、良好的灵敏度和高重现性。所提出的方法也是首次将 DBZD、BZD 和 Z 类催眠药的分析纳入一种高效分析的方法。
