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.

中文翻译：

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使用 QuEChERS 萃取和 LC-MS-MS 分析对肝脏组织中的 34 种芬太尼类似物进行定量和验证

自 2013 年以来，涉及合成阿片类药物（包括芬太尼和芬太尼类似物）的药物过量死亡人数从 3,105 人增加到 2018 年的 31,335 人。芬太尼相关药物过量死亡的尸检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以及与毒性、多药使用、新兴芬太尼类似物的新颖性和死后重新分配的关联。本研究的目标包括开发一种快速、简单、廉价、有效、耐用且安全的 (QuEChERS) 提取方法以及随后的液相色谱-质谱/质谱分析，并根据美国法医学会标准委员会验证该方法（ ASB) 标准 036 对 34 种分析物（包括芬太尼）的真实肝脏标本的要求和应用，代谢物和芬太尼类似物。对于任何低、中或高浓度，所有 34 种芬太尼类似物的偏差均不超过 ±10%，并且 %CV 不超过 20%。没有发现干扰。所有 34 种分析物均在可接受的电离百分比抑制或增强标准范围内，低浓度范围为 -10.2% 至 23.7%，高浓度范围为 -7.1% 至 11.0%。提取并分析了来自 22 个真实死后病例的肝脏样本，所有样本对 34 种化合物中的至少一种目标分析物均为阳性。在 22 个样品中，17 个含有芬太尼及其代谢物以及至少一种芬太尼类似物。芬太尼类似物的最高浓度为 541.4 μg/kg 对氟异丁酰芬太尼 (FIBF)。芬太尼 (n = 20) 的浓度范围在 3.6 和 164 之间。9 μg/kg，平均值为 54.7 μg/kg。最常见的芬太尼类似物是甲氧基乙酰芬太尼（n = 11），范围为 0.2-4.6 μg/kg，平均值为 1.3 μg/kg。使用 ASB 标准 036 对肝组织中芬太尼、代谢物和芬太尼类似物的要求对 QuEChERS 提取进行了全面验证。