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Quantitation of Fentanyl and Metabolites from Liver Tissue Using a Validated QuEChERS Extraction and LC-MS-MS Analysis.
J Anal Toxicol. 2021 Jan 21; 44(9):957-967.JA

Abstract

According to the National Institute on Drug Abuse (NIDA), more than one hundred people die every day from opioid overdose. Overdose fatalities have risen as the availability of potent synthetic opioids, such as fentanyl, has increased. A forensic postmortem toxicological specimen is often in various stages of decomposition, experiencing autolysis and putrefaction, which complicates the extraction, creating a difficult challenge for toxicologists. Isolating the target drug, while creating an efficient and simplified analytical scheme, is a goal for most toxicology laboratories. The validation of a quick, easy, cheap, effective, rugged and safe extraction protocol is presented in this study as an alternative analytical method for efficient extraction and detection of fentanyl and its major metabolites: norfentanyl and despropionyl fentanyl (4-ANPP). The liquid Chromatography with tandem mass spectrometry analysis was validated following the American Academy of Forensic Sciences Standards Board (ASB) standard 036 proposed requirements. Evaluated parameters include selectivity, matrix effects (MEs), linearity, processed sample stability, bias, precision and proof of applicability using liver samples from authentic postmortem cases. MEs (represented as percent ionization suppression or enhancement) at low and high concentrations were -10.0% and 1.4% for fentanyl, -2.1% and -0.3% for 4-ANPP and 3.1% and 2.8% for norfentanyl, respectively. Bias for the three analytes ranged from -8.5% to -19.9% for the low concentrations, -3.6% to -14.7% for the medium concentrations and 1.5% to -16.1% for the high concentrations with all being within the ±20% guideline. Precision for the three analytes ranged from 2.2% to 15.1%. The linear range for the fentanyl and norfentanyl was 0.5-100 and 4-ANPP had a linear range of 0.4-80 μg/kg. The authentic postmortem liver samples ranged in fentanyl concentrations from 56.6 to 462.3 μg/kg with a mean of 149.2 μg/kg (n = 10). The range of norfentanyl concentrations were 1.9 to 50.0 μg/kg with a mean of 14.1 μg/kg (n = 10). The range of 4-ANPP concentrations were 3.2 to 23.7 μg/kg with a mean of 7.5 μg/kg (n = 7).

Authors+Show Affiliations

Department of Forensic and Investigative Science, West Virginia University, 1600 University Avenue, Morgantown, WV 26506, USA.Department of Forensic and Investigative Science, West Virginia University, 1600 University Avenue, Morgantown, WV 26506, USA.Department of Forensic and Investigative Science, West Virginia University, 1600 University Avenue, Morgantown, WV 26506, USA.Department of Forensic and Investigative Science, West Virginia University, 1600 University Avenue, Morgantown, WV 26506, USA.Department of Pathology, Allegheny General Hospital, 320 E. North Avenue, Pittsburgh, PA 15212, USA.Office of the Chief Medical Examiner, 619 Virginia Street West, Charleston, WV 25302, USA.Office of the Chief Medical Examiner, 619 Virginia Street West, Charleston, WV 25302, USA.Office of the Chief Medical Examiner, 619 Virginia Street West, Charleston, WV 25302, USA.Department of Forensic and Investigative Science, West Virginia University, 1600 University Avenue, Morgantown, WV 26506, USA.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32020181

Citation

Cox, Joseph, et al. "Quantitation of Fentanyl and Metabolites From Liver Tissue Using a Validated QuEChERS Extraction and LC-MS-MS Analysis." Journal of Analytical Toxicology, vol. 44, no. 9, 2021, pp. 957-967.
Cox J, Train A, Field A, et al. Quantitation of Fentanyl and Metabolites from Liver Tissue Using a Validated QuEChERS Extraction and LC-MS-MS Analysis. J Anal Toxicol. 2021;44(9):957-967.
Cox, J., Train, A., Field, A., Ott, C., DelTondo, J., Kraner, J., Bailey, K., Gebhardt, M., & Arroyo-Mora, L. E. (2021). Quantitation of Fentanyl and Metabolites from Liver Tissue Using a Validated QuEChERS Extraction and LC-MS-MS Analysis. Journal of Analytical Toxicology, 44(9), 957-967. https://doi.org/10.1093/jat/bkaa006
Cox J, et al. Quantitation of Fentanyl and Metabolites From Liver Tissue Using a Validated QuEChERS Extraction and LC-MS-MS Analysis. J Anal Toxicol. 2021 Jan 21;44(9):957-967. PubMed PMID: 32020181.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Quantitation of Fentanyl and Metabolites from Liver Tissue Using a Validated QuEChERS Extraction and LC-MS-MS Analysis. AU - Cox,Joseph, AU - Train,Alex, AU - Field,Avery, AU - Ott,Colby, AU - DelTondo,Joseph, AU - Kraner,James, AU - Bailey,Kristen, AU - Gebhardt,Myron, AU - Arroyo-Mora,Luis E, PY - 2019/08/30/received PY - 2019/12/04/revised PY - 2019/12/07/accepted PY - 2020/2/6/pubmed PY - 2021/2/5/medline PY - 2020/2/6/entrez SP - 957 EP - 967 JF - Journal of analytical toxicology JO - J Anal Toxicol VL - 44 IS - 9 N2 - According to the National Institute on Drug Abuse (NIDA), more than one hundred people die every day from opioid overdose. Overdose fatalities have risen as the availability of potent synthetic opioids, such as fentanyl, has increased. A forensic postmortem toxicological specimen is often in various stages of decomposition, experiencing autolysis and putrefaction, which complicates the extraction, creating a difficult challenge for toxicologists. Isolating the target drug, while creating an efficient and simplified analytical scheme, is a goal for most toxicology laboratories. The validation of a quick, easy, cheap, effective, rugged and safe extraction protocol is presented in this study as an alternative analytical method for efficient extraction and detection of fentanyl and its major metabolites: norfentanyl and despropionyl fentanyl (4-ANPP). The liquid Chromatography with tandem mass spectrometry analysis was validated following the American Academy of Forensic Sciences Standards Board (ASB) standard 036 proposed requirements. Evaluated parameters include selectivity, matrix effects (MEs), linearity, processed sample stability, bias, precision and proof of applicability using liver samples from authentic postmortem cases. MEs (represented as percent ionization suppression or enhancement) at low and high concentrations were -10.0% and 1.4% for fentanyl, -2.1% and -0.3% for 4-ANPP and 3.1% and 2.8% for norfentanyl, respectively. Bias for the three analytes ranged from -8.5% to -19.9% for the low concentrations, -3.6% to -14.7% for the medium concentrations and 1.5% to -16.1% for the high concentrations with all being within the ±20% guideline. Precision for the three analytes ranged from 2.2% to 15.1%. The linear range for the fentanyl and norfentanyl was 0.5-100 and 4-ANPP had a linear range of 0.4-80 μg/kg. The authentic postmortem liver samples ranged in fentanyl concentrations from 56.6 to 462.3 μg/kg with a mean of 149.2 μg/kg (n = 10). The range of norfentanyl concentrations were 1.9 to 50.0 μg/kg with a mean of 14.1 μg/kg (n = 10). The range of 4-ANPP concentrations were 3.2 to 23.7 μg/kg with a mean of 7.5 μg/kg (n = 7). SN - 1945-2403 UR - https://www.unboundmedicine.com/medline/citation/32020181/Quantitation_of_Fentanyl_and_Metabolites_from_Liver_Tissue_Using_a_Validated_QuEChERS_Extraction_and_LC_MS_MS_Analysis_ L2 - https://academic.oup.com/jat/article-lookup/doi/10.1093/jat/bkaa006 DB - PRIME DP - Unbound Medicine ER -