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Long-Term Stability of 13 Fentanyl Analogs in Blood.
J Anal Toxicol. 2021 May 19 [Online ahead of print]JA

Abstract

Fentanyl analogs continue to play a major role in proliferating the opioid epidemic in the United States. With high rates of overdose deaths, forensic laboratories experience backlogs, which may lead to false negative results due to drug instability. To address this issue, a quantitative method was validated for fentanyl analogs (3-methylfentanyl, 4-ANPP, 4-fluoro-isobutyrylfentanyl (4-FIBF), acetylfentanyl, acrylfentanyl, butyrylfentanyl, carfentanil, cyclopropylfentanyl, fentanyl, furanylfentanyl, methoxyacetylfentanyl, p-fluorofentanyl, and valerylfentanyl) in blood using liquid chromatography-quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS) and used to assess long-term stability under various temperature conditions (-20°C, 4°C, ~25°C, and 35°C) for 9 months. Authentic specimens were also analyzed 6 months apart for applicability to postmortem blood. Method validation resulted in calibration ranges of 1-100 ng/mL and limits of detection (LOD) of 0.5 ng/mL. Precision and bias were acceptable (within ±7.2%CV and ±15.2%, respectively). Matrix effects exhibited ion enhancement for all analytes, except carfentanil and 4-ANPP in low quality control (>25%). For long-term stability, fentanyl analogs (except acrylfentanyl) remained stable under room temperature and refrigerated conditions at low and high concentrations (81.3-112.5% target) for 9 months. While most fentanyl analogs remained stable frozen, degradation was observed after 2 weeks (4 freeze/thaw cycles). At elevated temperatures, most analytes were stable for 1 week (74.2-112.6% target). Acrylfentanyl was unstable after 24h under elevated (70% loss) and room temperatures (53-60% loss), 48-72h refrigerated (28-40% loss), and 4 weeks frozen (22% loss). In authentic bloods (n=7), initial furanylfentanyl (FuF) and 4-ANPP concentrations were 1.1-3.6 and 1.4-6.4 ng/mL, respectively. Percent loss of FuF and 4-ANPP over 6 months were 16.3-37.4% and 0.2-26.8%, respectively. Samples suspected to contain fentanyl analogs are recommended to be stored refrigerated or frozen with limited freeze/thaw cycles. Due to instability, in the event of an acrylfentanyl overdose, samples should be analyzed immediately or stored frozen with analysis within 1 month.

Authors+Show Affiliations

Department of Forensic Science, College of Criminal Justice, Sam Houston State University, Huntsville, TX, USA.Department of Forensic Science, College of Criminal Justice, Sam Houston State University, Huntsville, TX, USA.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

34008837

Citation

Palmquist, Kaitlyn B., and Madeleine J. Swortwood. "Long-Term Stability of 13 Fentanyl Analogs in Blood." Journal of Analytical Toxicology, 2021.
Palmquist KB, Swortwood MJ. Long-Term Stability of 13 Fentanyl Analogs in Blood. J Anal Toxicol. 2021.
Palmquist, K. B., & Swortwood, M. J. (2021). Long-Term Stability of 13 Fentanyl Analogs in Blood. Journal of Analytical Toxicology. https://doi.org/10.1093/jat/bkab051
Palmquist KB, Swortwood MJ. Long-Term Stability of 13 Fentanyl Analogs in Blood. J Anal Toxicol. 2021 May 19; PubMed PMID: 34008837.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Long-Term Stability of 13 Fentanyl Analogs in Blood. AU - Palmquist,Kaitlyn B, AU - Swortwood,Madeleine J, Y1 - 2021/05/19/ PY - 2021/03/09/received PY - 2021/04/26/revised PY - 2021/05/18/accepted PY - 2021/5/19/entrez PY - 2021/5/20/pubmed PY - 2021/5/20/medline KW - Fentanyl analogs KW - LC–QTOF-MS KW - Stability JF - Journal of analytical toxicology JO - J Anal Toxicol N2 - Fentanyl analogs continue to play a major role in proliferating the opioid epidemic in the United States. With high rates of overdose deaths, forensic laboratories experience backlogs, which may lead to false negative results due to drug instability. To address this issue, a quantitative method was validated for fentanyl analogs (3-methylfentanyl, 4-ANPP, 4-fluoro-isobutyrylfentanyl (4-FIBF), acetylfentanyl, acrylfentanyl, butyrylfentanyl, carfentanil, cyclopropylfentanyl, fentanyl, furanylfentanyl, methoxyacetylfentanyl, p-fluorofentanyl, and valerylfentanyl) in blood using liquid chromatography-quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS) and used to assess long-term stability under various temperature conditions (-20°C, 4°C, ~25°C, and 35°C) for 9 months. Authentic specimens were also analyzed 6 months apart for applicability to postmortem blood. Method validation resulted in calibration ranges of 1-100 ng/mL and limits of detection (LOD) of 0.5 ng/mL. Precision and bias were acceptable (within ±7.2%CV and ±15.2%, respectively). Matrix effects exhibited ion enhancement for all analytes, except carfentanil and 4-ANPP in low quality control (>25%). For long-term stability, fentanyl analogs (except acrylfentanyl) remained stable under room temperature and refrigerated conditions at low and high concentrations (81.3-112.5% target) for 9 months. While most fentanyl analogs remained stable frozen, degradation was observed after 2 weeks (4 freeze/thaw cycles). At elevated temperatures, most analytes were stable for 1 week (74.2-112.6% target). Acrylfentanyl was unstable after 24h under elevated (70% loss) and room temperatures (53-60% loss), 48-72h refrigerated (28-40% loss), and 4 weeks frozen (22% loss). In authentic bloods (n=7), initial furanylfentanyl (FuF) and 4-ANPP concentrations were 1.1-3.6 and 1.4-6.4 ng/mL, respectively. Percent loss of FuF and 4-ANPP over 6 months were 16.3-37.4% and 0.2-26.8%, respectively. Samples suspected to contain fentanyl analogs are recommended to be stored refrigerated or frozen with limited freeze/thaw cycles. Due to instability, in the event of an acrylfentanyl overdose, samples should be analyzed immediately or stored frozen with analysis within 1 month. SN - 1945-2403 UR - https://www.unboundmedicine.com/medline/citation/34008837/Long_Term_Stability_of_13_Fentanyl_Analogs_in_Blood_ L2 - https://academic.oup.com/jat/article-lookup/doi/10.1093/jat/bkab051 DB - PRIME DP - Unbound Medicine ER -
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