Abstract

Fentanyl and its analogues are highly abused drugs that dominate the illicit drug trade. alpha-Methylfentanyl (A-F) and beta-hydroxyfentanyl (B-F) are two fentanyl analogues that require the development of rapid detection technologies. The current study established and validated a rapid and high-sensitivity liquid chromatography–tandem mass spectrometry (LC–MS-MS) method to measure A-F and B-F concentrations in rat plasma following intravenous drug administration (20 μg/kg). Because fentanyl is primarily metabolized by the liver, we evaluated the concentrations of A-F and B-F in vivo in rats, in a control group and a group with liver damage induced by 55 days of oral ethanol gavage (6.5 g/kg, 22.5% v/v). Liquid–liquid extraction and LC–MS-MS operating in the positive ion multiple reaction monitoring mode were used. A C₁₈ column was used, and the mobile phase consisted of 0.1% formic acid aqueous and acetonitrile. The limit of detection was 3 pg/mL (S/N > 5) for A-F and B-F. The calibration curves were linear within the concentration range of 0.01–5 ng/mL (R² = 0.9991) and 0.005–20 ng/mL (R² = 0.9999) for A-F and B-F, respectively. Extraction recoveries were 91.3%–97.6% with RSD ≤ 11.2% and 90.5%–94.3% with RSD ≤ 10.5% for A-F and B-F, respectively. Plasma matrix effects were 80.61%–84.58% for A-F and 80.67%–81.33% for B-F with RSD ≤ 13.9%. The validated assay indicated no significant differences in pharmacokinetic parameters (AUC_0-t, C_max and T_1/2) derived from the assessment of A-F and B-F plasma concentrations between control and ethanol-exposed rats. This assay, for which the LOD was 3 pg/mL for A-F and B-F may help the forensic science field to determine fentanyl analogue-related causes of death and identify illicit drug tampering.

中文翻译：

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长期暴露于乙醇后大鼠血浆中的α-甲基芬太尼和β-羟基芬太尼LC-MS-MS定量

摘要

芬太尼及其类似物是滥用毒品，在非法毒品贸易中占主导地位。α-甲基芬太尼（AF）和β-羟基芬太尼（BF）是需要快速检测技术发展的两种芬太尼类似物。当前的研究建立并验证了一种快速，高灵敏度的液相色谱-串联质谱法（LC-MS-MS），用于在静脉给药后测量大鼠血浆中的AF和BF浓度（20μg/ kg）。由于芬太尼主要通过肝脏代谢，因此我们评估了体内AF和BF的浓度在大鼠，对照组和55天口服乙醇灌胃（6.5 g / kg，22.5％v / v）引起的肝损伤组中。使用在正离子多反应监测模式下运行的液-液萃取和LC-MS-MS。使用AC ₁₈柱，流动相由0.1％的甲酸水溶液和乙腈组成。AF和BF的检出限为3 pg / mL（S / N> 5）。浓度范围为0.01–5 ng / mL（R ²  = 0.9991）和0.005–20 ng / mL（R ² = 0.9999），分别适用于AF和BF。AF和BF的萃取回收率分别为91.3％–97.6％（RSD≤11.2％）和90.5％–94.3％（RSD≤10.5％）。AF和RSD≤13.9％的高炉血浆基质效应分别为80.61％–84.58％和80.67％–81.33％。经过验证的分析表明，对照组和乙醇暴露大鼠的AF和BF血浆浓度评估得出的药代动力学参数（AUC _0-t，C _max和T _1/2）没有显着差异。AF和BF的LOD为3 pg / mL的这种​​测定法可能有助于法医科学领域确定与芬太尼类似物相关的死亡原因并鉴定非法药物篡改。