Department of Forensic and Investigative Science, 1600 University Avenue, Oglebay Hall, Room 302, Morgantown, WV 26506, USA.
Department of Pathology, Allegheny General Hospital, 320 E North Avenue, Pittsburgh, PA 15212, USA.
J Anal Toxicol. 2022 Mar 21;46(3):232-245. doi: 10.1093/jat/bkab009.
Since 2013, drug overdose deaths involving synthetic opioids (including fentanyl and fentanyl analogs) have increased from 3,105 to 31,335 in 2018. Postmortem toxicological analysis in fentanyl-related overdose deaths is complicated by the high potency of the drug, often resulting in low analyte concentrations and associations with toxicity, multidrug use, novelty of emerging fentanyl analogs and postmortem redistribution. Objectives for this study include the development of a quick, easy, cheap, effective, rugged and safe (QuEChERS) extraction and subsequent liquid chromatography-mass spectrometry--mass spectrometry analysis, validation of the method following the American Academy of Forensic Sciences Standards Board (ASB) standard 036 requirements and application to authentic liver specimens for 34 analytes including fentanyl, metabolites and fentanyl analogs. The bias for all 34 fentanyl analogs did not exceed ±10% for any of the low, medium or high concentrations and the %CV did not exceed 20%. No interferences were identified. All 34 analytes were within the criteria for acceptable percent ionization suppression or enhancement with the low concentration ranging from -10.2% to 23.7% and the high concentration ranging from -7.1% to 11.0%. Liver specimens from 22 authentic postmortem cases were extracted and analyzed with all samples being positive for at least one target analyte from the 34 compounds. Of the 22 samples, 17 contained fentanyl and metabolites plus at least one fentanyl analog. The highest concentration for a fentanyl analog was 541.4 μg/kg of para-fluoroisobutyryl fentanyl (FIBF). The concentrations for fentanyl (n = 20) ranged between 3.6 and 164.9 μg/kg with a mean of 54.7 μg/kg. The fentanyl analog that was most encountered was methoxyacetyl fentanyl (n = 11) with a range of 0.2-4.6 μg/kg and a mean of 1.3 μg/kg. The QuEChERS extraction was fully validated using the ASB Standard 036 requirements for fentanyl, metabolites and fentanyl analogs in liver tissue.
自 2013 年以来，涉及合成阿片类药物（包括芬太尼及其类似物）的药物过量死亡人数从 2013 年的 3105 人增加到 2018 年的 31335 人。芬太尼相关药物过量死亡的死后毒理学分析因药物的高效力而变得复杂，通常导致分析物浓度低，并与毒性、多药使用、新兴芬太尼类似物的新颖性和死后再分布有关。本研究的目的包括开发一种快速、简便、廉价、有效、坚固和安全（QuEChERS）的提取方法，随后进行液相色谱-质谱-质谱分析，按照美国法医科学协会标准委员会（ASB）标准 036 的要求对方法进行验证，并应用于 34 种分析物（包括芬太尼、代谢物和芬太尼类似物）的真实肝脏标本。所有 34 种芬太尼类似物的偏差均不超过低、中、高浓度任何一种的±10%，CV 不超过 20%。未发现干扰。所有 34 种分析物均符合可接受的百分离子化抑制或增强标准，低浓度范围为-10.2%至 23.7%，高浓度范围为-7.1%至 11.0%。从 22 例真实死后案例中提取并分析了肝脏标本，所有样本均至少有一种来自 34 种化合物的目标分析物呈阳性。在 22 个样本中，17 个样本中含有芬太尼及其代谢物，再加上至少一种芬太尼类似物。芬太尼类似物的最高浓度为 541.4μg/kg 对氟异丁酰芬太尼（FIBF）。芬太尼（n=20）的浓度范围在 3.6 至 164.9μg/kg 之间，平均值为 54.7μg/kg。最常见的芬太尼类似物是甲氧基乙酰芬太尼（n=11），浓度范围为 0.2-4.6μg/kg，平均值为 1.3μg/kg。QuEChERS 提取方法完全按照 ASB 标准 036 对肝脏组织中的芬太尼、代谢物和芬太尼类似物的要求进行了验证。
