National Institute of Legal Medicine and Forensic Sciences, South Branch, Rua Manuel Bento de Sousa n°3, 1169-201 Lisbon, Portugal.
National Institute of Legal Medicine and Forensic Sciences, South Branch, Rua Manuel Bento de Sousa n°3, 1169-201 Lisbon, Portugal.
J Pharm Biomed Anal. 2018 Jan 5;147:634-644. doi: 10.1016/j.jpba.2017.02.046. Epub 2017 Feb 27.
A method for the simultaneous determination of 11 illicit drugs, using the dried blood spot (DBS) sampling technique combined with the UPLC-MS/MS technology was developed to study its applicability within the forensic toxicology. The DBS samples, prepared from a blood volume of 50μL and using the Whatman® BFC 180 bloodstain cards, were extracted with a methanol/acetonitrile mixture. The chromatographic separation was performed using an Acquity UPLC HSS T3 column (100mm×2.1mm, 1.8μm) and an acetonitrile/2mM ammonium formate (0.1% formic acid) gradient. The detection was accomplished with a TQ Detector, operating in the ESI+ and MRM modes. The method was validated in terms of selectivity, matrix effect, extraction recovery (42%-91%), carryover, LOD and LOQ (0.5-1ng/mL and 1-5ng/mL, respectively), linearity (LOQ to 500ng/mL), intraday and interday precision (3.8-14% and 5.3-13%, respectively), accuracy (-9.3% to 7.9%) and dilution integrity. An eight months stability study at room temperature, 2-8°C and -10°C, was also performed, with the best results obtained at -10°C. The procedure was applied to 64 real samples (92 positive results for substances included in this study). The results were compared with the methodologies routinely applied in the laboratory and the statistical analysis allowed to establish an acceptable correlation. This study permitted to determine that the DBS can represent an alternative or a complement to conventional analytical and sampling techniques, responding to some of the present issues concerning the different forensic toxicology applications.
建立了一种同时测定 11 种非法药物的方法，采用干血斑（DBS）采样技术结合 UPLC-MS/MS 技术，研究其在法医毒理学中的适用性。DBS 样品由 50μL 血液体积制备，使用 Whatman® BFC 180 血斑卡，用甲醇/乙腈混合物提取。采用 Acquity UPLC HSS T3 柱（100mm×2.1mm，1.8μm）和乙腈/2mM 甲酸铵（0.1%甲酸）梯度进行色谱分离。采用 TQ 检测器，在 ESI+和 MRM 模式下进行检测。该方法在选择性、基质效应、提取回收率（42%-91%）、携带污染、LOD 和 LOQ（0.5-1ng/mL 和 1-5ng/mL）、线性（LOQ 至 500ng/mL）、日内和日间精密度（3.8%-14%和 5.3%-13%）、准确度（-9.3%至 7.9%）和稀释完整性方面进行了验证。还进行了为期八个月的室温、2-8°C 和-10°C 稳定性研究，-10°C 时获得的结果最佳。该程序应用于 64 个实际样本（包括本研究中物质的 92 个阳性结果）。将结果与实验室常规应用的方法进行比较，统计分析允许建立可接受的相关性。这项研究表明，DBS 可以替代或补充传统的分析和采样技术，解决法医毒理学应用中存在的一些问题。
