Institute of Forensic Medicine, Forensic Toxicology, Kantonsspital St. Gallen, St. Gallen 9007, Switzerland.
Anal Chem. 2023 Feb 7;95(5):2723-2731. doi: 10.1021/acs.analchem.2c03516. Epub 2023 Jan 27.
Consumption of opioids is a growing global health problem. The gold standard for drugs of abuse screening is immunochemical assays. However, this method comes with some disadvantages when screening for a wide variety of opioids. Detection of the binding of a compound at the human μ-opioid receptor (MOR) offers a promising alternative target. Here, we set up a urine assay to allow for detection of compounds that bind at the MOR, thus allowing the assay to be utilized as a screening tool for opioid intake. The assay is based on the incubation of MOR-containing cell membranes with the selective MOR-ligand DAMGO and urine. After filtration, the amount of DAMGO in the eluate is analyzed by liquid chromatography tandem mass spectroscopy (LC-MS/MS). The absence of DAMGO in the eluate corresponds to a competing MOR ligand in the urine sample, thus indicating opiate/opioid intake by the suspect. Sensitivity and specificity were determined by the analysis of 200 consecutive forensic routine casework urine samples. A pronounced displacement of DAMGO was observed in 29 of the 35 opiate/opioid-positive samples. Detection of fentanyl intake proved to be the most challenging aspect. Applying a cut-off value of, e.g., 10% DAMGO binding would lead to a sensitivity of 83% and a specificity of 95%. Consequently, the novel assay proved to be a promising screening tool for opiate/opioid presence in urine samples. The nontargeted approach and possible automation of the assay make it a promising alternative to conventional methods.
阿片类药物的消费是一个日益严重的全球健康问题。滥用药物筛选的金标准是免疫化学分析。然而，当筛选各种阿片类药物时，这种方法存在一些缺点。检测化合物与人μ-阿片受体（MOR）的结合提供了一个有前途的替代靶标。在这里，我们建立了一种尿液分析方法，以允许检测与 MOR 结合的化合物，从而使该分析方法可作为阿片类药物摄入的筛选工具。该分析方法基于在含有 MOR 的细胞膜与选择性 MOR 配体 DAMGO 和尿液孵育。过滤后，通过液相色谱串联质谱（LC-MS/MS）分析洗脱液中 DAMGO 的含量。洗脱液中不存在 DAMGO 对应于尿液样品中竞争 MOR 配体的物质，从而表明嫌疑犯摄入了阿片类药物/阿片类物质。通过分析 200 个连续的法医常规案例尿液样本确定了灵敏度和特异性。在 35 个阿片类药物/阿片类药物阳性样本中观察到 DAMGO 明显置换。检测芬太尼摄入被证明是最具挑战性的方面。例如，应用 10% DAMGO 结合的截止值将导致灵敏度为 83%，特异性为 95%。因此，新的分析方法被证明是一种有前途的尿液样本中阿片类药物/阿片类物质存在的筛选工具。该非靶向方法和分析的可能自动化使其成为传统方法的有前途的替代方法。
