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Validation and Cross-Reactivity Data for Fentanyl Analogs With the Immunalysis Fentanyl ELISA.
J Anal Toxicol. 2019 Jan 01; 43(1):18-24.JA

Abstract

Every year new fentanyl analog compounds, or fentanyls, appear on the drug scene. Development of immunoassays dedicated for screening individual molecules is challenging due to the short-lived presence of these compounds on the recreational drug market. Therefore, we investigated the detecting capabilities of the immunalysis fentanyl direct enzyme-linked immunosorbent assay (ELISA) kit against fentanyl in whole blood, and determined the cross-reactivity of nine fentanyl analogs (2-fluorofentanyl, acetylfentanyl, acrylfentanyl, carfentanil, cyclopropylfentanyl, tetrahydrofuranylfentanyl, furanylfentanyl, ocfentanil, valerylfentanyl) to confirm its validity for the general screening of fentanyls. Immunalysis ELISA assay was used to test whole blood samples fortified with fentanyl on a TECAN Freedom EVOlyzer platform, according to manufacturer specifications. The kit successfully was validated for fentanyl screening with a cutoff set at 0.5 ng/mL, and all tested analogs, with the exclusion of carfentanil, were detected. The lowest cross-reactivity with the kit was obtained with furanylfentanyl (20% ± 1, 95% confidence intervals (CI)) and 4-fluoroisobutyrfentanyl (25% ± 1, 95% CI), while the highest was recorded using acetylfentanyl (99% ± 11, 95% CI) and acrylfentanyl (94% ± 10, 95% CI). Post-mortem samples containing fentanyl, acrylfentanyl, cyclopropylfentanyl, THF-fentanyl and 4-fluoroisobutyrfentanyl were screened, and sensitivity and specificity of each analog were calculated. Positive screening results were generated by all post-mortem cases containing fentanyl (n = 14), acrylfentanyl (n = 11), cyclopropylfentanyl (n = 14), tetrahydrofuranylfentanyl (n = 13) and 4-fluoroisobutyrfentanyl (n = 10). Concentration of post-mortem fentanyl samples ranged from 0.5 ng/mL (cutoff) to 230 ng/mL, while the range for analogs was 3.4-36 ng/mL (cyclopentylfentanyl), 0.76-370 ng/mL (4-fluoroisobutyrfentanyl), 0.02-12 ng/mL (acrylfentanyl) and 2-26 ng/mL (tetrahydrofuranylfentanyl). The immunalysis fentanyl direct ELISA kit was successfully validated and showed significant cross-reactivity for all tested fentanyls, except carfentanil, making it a suitable technique for fentanyl and fentanyl analogs screening.

Authors+Show Affiliations

Department of Medical and Health Science, Division of Drug Research, Linköping University, Linköping, Sweden.Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Linköping, Sweden.Department of Medical and Health Science, Division of Drug Research, Linköping University, Linköping, Sweden. Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Linköping, Sweden.Department of Medical and Health Science, Division of Drug Research, Linköping University, Linköping, Sweden. Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Linköping, Sweden.

Pub Type(s)

Journal Article
Validation Study

Language

eng

PubMed ID

30215771

Citation

Guerrieri, Davide, et al. "Validation and Cross-Reactivity Data for Fentanyl Analogs With the Immunalysis Fentanyl ELISA." Journal of Analytical Toxicology, vol. 43, no. 1, 2019, pp. 18-24.
Guerrieri D, Kjellqvist F, Kronstrand R, et al. Validation and Cross-Reactivity Data for Fentanyl Analogs With the Immunalysis Fentanyl ELISA. J Anal Toxicol. 2019;43(1):18-24.
Guerrieri, D., Kjellqvist, F., Kronstrand, R., & Gréen, H. (2019). Validation and Cross-Reactivity Data for Fentanyl Analogs With the Immunalysis Fentanyl ELISA. Journal of Analytical Toxicology, 43(1), 18-24. https://doi.org/10.1093/jat/bky060
Guerrieri D, et al. Validation and Cross-Reactivity Data for Fentanyl Analogs With the Immunalysis Fentanyl ELISA. J Anal Toxicol. 2019 Jan 1;43(1):18-24. PubMed PMID: 30215771.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Validation and Cross-Reactivity Data for Fentanyl Analogs With the Immunalysis Fentanyl ELISA. AU - Guerrieri,Davide, AU - Kjellqvist,Fanny, AU - Kronstrand,Robert, AU - Gréen,Henrik, PY - 2018/03/02/received PY - 2018/9/15/pubmed PY - 2019/3/15/medline PY - 2018/9/15/entrez SP - 18 EP - 24 JF - Journal of analytical toxicology JO - J Anal Toxicol VL - 43 IS - 1 N2 - Every year new fentanyl analog compounds, or fentanyls, appear on the drug scene. Development of immunoassays dedicated for screening individual molecules is challenging due to the short-lived presence of these compounds on the recreational drug market. Therefore, we investigated the detecting capabilities of the immunalysis fentanyl direct enzyme-linked immunosorbent assay (ELISA) kit against fentanyl in whole blood, and determined the cross-reactivity of nine fentanyl analogs (2-fluorofentanyl, acetylfentanyl, acrylfentanyl, carfentanil, cyclopropylfentanyl, tetrahydrofuranylfentanyl, furanylfentanyl, ocfentanil, valerylfentanyl) to confirm its validity for the general screening of fentanyls. Immunalysis ELISA assay was used to test whole blood samples fortified with fentanyl on a TECAN Freedom EVOlyzer platform, according to manufacturer specifications. The kit successfully was validated for fentanyl screening with a cutoff set at 0.5 ng/mL, and all tested analogs, with the exclusion of carfentanil, were detected. The lowest cross-reactivity with the kit was obtained with furanylfentanyl (20% ± 1, 95% confidence intervals (CI)) and 4-fluoroisobutyrfentanyl (25% ± 1, 95% CI), while the highest was recorded using acetylfentanyl (99% ± 11, 95% CI) and acrylfentanyl (94% ± 10, 95% CI). Post-mortem samples containing fentanyl, acrylfentanyl, cyclopropylfentanyl, THF-fentanyl and 4-fluoroisobutyrfentanyl were screened, and sensitivity and specificity of each analog were calculated. Positive screening results were generated by all post-mortem cases containing fentanyl (n = 14), acrylfentanyl (n = 11), cyclopropylfentanyl (n = 14), tetrahydrofuranylfentanyl (n = 13) and 4-fluoroisobutyrfentanyl (n = 10). Concentration of post-mortem fentanyl samples ranged from 0.5 ng/mL (cutoff) to 230 ng/mL, while the range for analogs was 3.4-36 ng/mL (cyclopentylfentanyl), 0.76-370 ng/mL (4-fluoroisobutyrfentanyl), 0.02-12 ng/mL (acrylfentanyl) and 2-26 ng/mL (tetrahydrofuranylfentanyl). The immunalysis fentanyl direct ELISA kit was successfully validated and showed significant cross-reactivity for all tested fentanyls, except carfentanil, making it a suitable technique for fentanyl and fentanyl analogs screening. SN - 1945-2403 UR - https://www.unboundmedicine.com/medline/citation/30215771/Validation_and_Cross_Reactivity_Data_for_Fentanyl_Analogs_With_the_Immunalysis_Fentanyl_ELISA_ L2 - https://academic.oup.com/jat/article-lookup/doi/10.1093/jat/bky060 DB - PRIME DP - Unbound Medicine ER -