Centro de Investigação em Ciências da Saúde (CICS-UBI), Universidade da Beira Interior, Av. Infante D. Henrique, 6201-556, Covilhã, Portugal.
Laboratório de Fármaco-Toxicologia, UBIMedical, Universidade da Beira Interior, Convento de Sto. António, 6201-001, Covilhã, Portugal.
Anal Bioanal Chem. 2018 Dec;410(30):7955-7964. doi: 10.1007/s00216-018-1417-8. Epub 2018 Nov 7.
This work describes the optimization of a new method for the determination of five organophosphorus insecticides in whole blood. The analytes were extracted from the matrix (50 μL) using the dried blood spot (DBS) approach and were analyzed by gas chromatography (GC) coupled to tandem mass spectrometry (MS/MS). The studied compounds (diazinon, chlorpyrifos, parathion-ethyl, chlorfenvinphos and quinalphos) were chosen based on the statistics of intoxications in Portugal, and ethion was used as internal standard. The method was fully validated, taking into account international guidelines for bioanalytical method validation, such as those of the Food and Drug Administration (FDA), International Conference on Harmonization (ICH) and Scientific Working Group for Forensic Toxicology (SWGTOX). A linear range of 0.1-25 μg/mL was obtained for all compounds, except for diazinon and quinalphos (0.05-25 μg/mL and 0.25-25 μg/mL, respectively), presenting determination coefficients above 0.99. Concerning precision, the coefficients of variation (CVs) were lower than 14% for all compounds. Those compounds were found to be stable in the samples. Although the values obtained for recovery were low (between 1 and 12%), the method proved to be sensitive, since detection limits between 0.05 and 0.1 μg/mL were obtained. The novelty is the use of the DBS approach in the extraction of these compounds, and this is the first paper reporting it: DBS is a recent technique of bioanalysis in the field of toxicology, and in addition to its simplicity and sensitivity, it is applicable routinely in both clinical and forensic toxicology situations. Graphical abstract ᅟ.
这项工作描述了一种新方法测定全血中五种有机磷杀虫剂的优化。采用干血斑(DBS)法从基质(50μL)中提取分析物,并用气相色谱(GC)串联质谱(MS/MS)进行分析。选择研究化合物(二嗪农、毒死蜱、乙基对硫磷、氯芬磷和喹硫磷)是基于葡萄牙中毒统计数据,并用乙硫磷作为内标。该方法是完全验证的,考虑到生物分析方法验证的国际指南,如美国食品和药物管理局(FDA)、国际协调会议(ICH)和法医毒理学科学工作组(SWGTOX)的指南。除二嗪农和喹硫磷(0.05-25μg/mL 和 0.25-25μg/mL)外,所有化合物的线性范围均为 0.1-25μg/mL,测定系数均大于 0.99。关于精密度,所有化合物的变异系数(CVs)均低于 14%。这些化合物在样品中稳定。虽然回收率值较低(在 1%到 12%之间),但该方法证明是敏感的,因为检测限在 0.05 到 0.1μg/mL 之间。新颖之处在于采用 DBS 法提取这些化合物,这是首次报道:DBS 是毒理学领域生物分析的一项新技术,除了简单和敏感之外,它还适用于临床和法医毒理学情况的常规应用。