Florou Dimitra, Orfanidis Amvrosios, Boumba Vassiliki A
Department of Forensic Medicine and Toxicology, Faculty of Medicine, School of Health Sciences, University of Ioannina, University Campus, Ioannina, 45500, Greece.
Department of Forensic Medicine and Toxicology, Faculty of Medicine, School of Health Sciences, University of Ioannina, University Campus, Ioannina, 45500, Greece; University Hospital of Ioannina, Ioannina, 45110, Greece.
J Chromatogr B Analyt Technol Biomed Life Sci. 2025 Nov 1;1265:124750. doi: 10.1016/j.jchromb.2025.124750. Epub 2025 Aug 7.
Urine toxicological analysis serves as a significant tool in both clinical and forensic contexts, facilitating the diagnosis of acute intoxications, determination of causes of death, monitoring of substance use in occupational settings, and identification of drug-facilitated crimes. In this regard, the dilute-and-shoot method, when integrated with liquid chromatography-tandem mass spectrometry (LC-MS/MS), represents a promising analytical approach due to its efficacy, reliability, and wide-ranging applicability. This study presents the development and validation of an LC-MS/MS method that simultaneously detects and quantitates 115 drugs and metabolites comprising analytes from different categories, such as drugs of abuse (DOA), new psychoactive substances (NPS), prescription and over-the-counter drugs. Sample pretreatment was studied by applying different solvents (acetonitrile, water or methanol), and the best results were obtained after adding 100 μL of sample, 200 μL of a mixture of methanol: acetonitrile (3:1, v/v). The mixture was vortexed, centrifuged and the supernatant directly injected into the instrument. The analysis took place on a C18 column with a gradient elution over 7.5 min. The method was found to be selective and sensitive, offering LOD/LOQ ranging from 0.01 to 1.5/ 0.05-5 ng/mL, respectively. Validation of the method included evaluation of recovery, carryover, matrix effect, accuracy, precision, selectivity, stability and dilution integrity. The method performed satisfactorily and was therefore applied to urine samples that collected over a 12-week period from individuals enrolled in a rehabilitation program. The proposed method was applied as a follow-up tool aiming to detect prescribed (or not) medicine, as well as other illegal substances.
尿液毒理学分析在临床和法医领域都是一项重要工具，有助于急性中毒的诊断、死因判定、职业环境中物质使用情况的监测以及药物辅助犯罪的识别。在这方面，稀释并注射法与液相色谱 - 串联质谱法（LC-MS/MS）相结合时，因其有效性、可靠性和广泛的适用性，是一种很有前景的分析方法。本研究介绍了一种LC-MS/MS方法的开发与验证，该方法可同时检测和定量115种药物及代谢物，这些分析物包括不同类别，如滥用药物（DOA）、新型精神活性物质（NPS）、处方药和非处方药。通过应用不同溶剂（乙腈、水或甲醇）研究了样品预处理，加入100 μL样品、200 μL甲醇与乙腈的混合物（3:1，v/v）后获得了最佳结果。将混合物涡旋、离心，然后将上清液直接注入仪器。分析在C18柱上进行，梯度洗脱7.5分钟。该方法具有选择性和灵敏性，检测限/定量限分别为0.01至1.5/0.05 - 5 ng/mL。该方法的验证包括回收率、残留、基质效应、准确性、精密度、选择性、稳定性和稀释完整性评估。该方法表现令人满意，因此应用于从参加康复计划的个体收集的为期12周的尿液样本。所提出的方法作为一种后续工具，旨在检测已开（或未开）药物以及其他非法物质。
