Laboratory for Forensic Chemistry, Department of Analytical Chemistry, Faculty of Chemistry, Jagiellonian University, 2, Gronostajowa St., 30-387 Kraków, Poland.
Laboratory for Forensic Chemistry, Department of Analytical Chemistry, Faculty of Chemistry, Jagiellonian University, 2, Gronostajowa St., 30-387 Kraków, Poland.
J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Nov 10;1157:122339. doi: 10.1016/j.jchromb.2020.122339. Epub 2020 Aug 21.
The aim of this study was to develop a new approach to sample preparation of biological material based on a combination of the Dried Blood Spot (DBS) method and capillary electrophoresis coupled with mass spectrometry (CE-MS) for the analysis of blood samples collected in vivo or post-mortem. The proposed approach allowed the identification of typical drugs from different groups, such as tricyclic antidepressants (amitriptyline, imipramine), selective serotonin reuptake inhibitors (citalopram), benzodiazepines (tetrazepam) and hypnotics (zolpidem). In this study, a blood sample was spotted on FTA DMPK C cards, then dried, and 6-mm discs were cut out. The sample preparation procedure involved microwave-assisted extraction (MAE). Various extraction agents, temperatures and durations of extraction were examined in order to achieve the highest efficiency of the process. The method was subjected to a validation procedure. Limits of detection (LOD = 1.76 - 14.7 ng/mL) and quantification (LOQ = 5.25 - 49.0 ng/mL), inter- (CV = 1.31 - 9.43%) and intra- (CV = 3.26 - 18.52%) day precision of the determinations, recovery (RE = 85.0-105.4%) and matrix effect on ionization of analytes (ME = 98.6-105.5%) were determined. Furthermore, the developed DBS/MAE/CM-MS method was selective and analytes present in the blood applied on DBS cards were found to be stable after 7 and after 14 days. Moreover, the developed method was successfully applied to the analysis of both post-mortem samples and blood samples taken from patients treated with the analyzed drugs.
本研究旨在开发一种新的生物材料样品制备方法,该方法基于干血斑 (DBS) 方法和毛细管电泳与质谱联用 (CE-MS) 的结合,用于分析体内或死后采集的血液样本。所提出的方法允许鉴定来自不同组的典型药物,例如三环抗抑郁药(阿米替林、丙咪嗪)、选择性 5-羟色胺再摄取抑制剂(西酞普兰)、苯并二氮䓬类(地西泮)和催眠药(唑吡坦)。在这项研究中,将血液样本点在 FTA DMPK C 卡上,然后干燥,然后切出 6-mm 圆盘。样品制备过程涉及微波辅助提取 (MAE)。研究了各种提取剂、提取温度和提取时间,以达到最高的过程效率。该方法经过验证程序。检测限 (LOD=1.76-14.7ng/mL) 和定量限 (LOQ=5.25-49.0ng/mL)、日内 (CV=1.31-9.43%) 和日间 (CV=3.26-18.52%) 精密度、回收率 (RE=85.0-105.4%) 和基质对分析物电离的影响 (ME=98.6-105.5%)。此外,开发的 DBS/MAE/CM-MS 方法具有选择性,并且应用于 DBS 卡上的血液中的分析物在 7 天和 14 天后被发现稳定。此外,该方法成功应用于分析死后样本和接受分析药物治疗的患者的血液样本。