Department of Forensic Science, Sam Houston State University, 1905 University Ave, Huntsville, TX 77340, USA.
J Anal Toxicol. 2020 Jul 31;44(6):589-595. doi: 10.1093/jat/bkaa013.
Fentanyl analogs (novel and traditional) continue to impact the ever-growing opioid epidemic. Furanylfentanyl (FuF) is one analog equipotent to fentanyl that has documented involvement in thousands of intoxication and fatality cases around the world. Due to its prevalence, toxicologists need to improve detection and understanding of this analog. A method for the quantification of FuF and its metabolites (4-ANPP, furanyl norfentanyl (FuNorF)) in a small volume (100 μL) of human plasma by LC-MS-MS was developed and validated according to ANSI/ASB Standard. The method was cross validated in rat plasma for a future pharmacokinetic (PK)/pharmacodynamic (PD) study. In human plasma, calibration ranges were 0.025-25 ng/mL (FuF and 4-ANPP) and 0.5-25 ng/mL (FuNorF). Limits of detection were 0.0125 ng/mL (FuF and 4-ANPP) and 0.25 ng/mL (FuNorF). Lower limits of quantification coincided with lowest calibrator concentrations of 0.025 ng/mL (FuF and 4-ANPP) and 0.5 ng/mL (FuNorF). Precision and bias values were determined to be acceptable for all analytes. Matrix effects were acceptable for all analytes (-8.6-25.0%), except FuNorF with suppression >25%. Extraction recoveries ranged from 84.5 to 98.1%. No carryover or endogenous interferences were observed. Qualitative interferences with 4-ANPP were observed from some n-acyl substituted fentanyl analogs predicted to be low-concentration standard impurities. Analytes were stable under all conditions and dilution integrity was sustained. The method was successfully cross validated in rat plasma with acceptable bias (-7.4-8.4%), precision (within-run < 19%CV and between-run < 12.6%CV), matrix effects (-9.3-17.2%, except FuNorF with >25% suppression), recoveries (79.2-94.5%) and dilution integrity (1/2 and 1/10).
芬太尼类似物（新型和传统）继续影响着日益严重的阿片类药物泛滥问题。呋喃芬太尼（FuF）是一种与芬太尼等效的类似物，在全球范围内有数千例中毒和死亡病例与它有关。由于其普遍存在，毒理学家需要提高对这种类似物的检测和理解。本文建立并验证了一种通过 LC-MS-MS 从小体积（100 μL）人血浆中定量检测 FuF 及其代谢物（4-ANPP、呋喃去甲芬太尼（FuNorF））的方法，该方法符合 ANSI/ASB 标准。该方法在大鼠血浆中进行了交叉验证，以便进行未来的药代动力学（PK）/药效动力学（PD）研究。在人血浆中，校准范围为 0.025-25ng/mL（FuF 和 4-ANPP）和 0.5-25ng/mL（FuNorF）。检测限为 0.0125ng/mL（FuF 和 4-ANPP）和 0.25ng/mL（FuNorF）。定量下限与最低校准浓度 0.025ng/mL（FuF 和 4-ANPP）和 0.5ng/mL（FuNorF）一致。所有分析物的精密度和偏差值均符合要求。除 FuNorF 有超过 25%的抑制作用外，所有分析物的基质效应均在可接受范围内（-8.6-25.0%）。萃取回收率为 84.5-98.1%。未观察到携带污染或内源性干扰。4-ANPP 与一些预测为低浓度标准杂质的 N-酰取代芬太尼类似物发生定性干扰。在所有条件下分析物均稳定，稀释完整性得以维持。该方法在大鼠血浆中成功进行了交叉验证，偏差（-7.4-8.4%）、精密度（批内<19%CV，批间<12.6%CV）、基质效应（-9.3-17.2%，FuNorF 除外，有超过 25%的抑制作用）、回收率（79.2-94.5%）和稀释完整性（1/2 和 1/10）均符合要求。
