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4 protocols using furanyl fentanyl

1

Opioid and Synthetic Opioid Separations

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The run buffer used for opioid and synthetic opioid separations was 12 mmol/L acetic acid, 3.3 mmol/L ammonium acetate pH 4.4 (Sigma-Aldrich, St. Louis, MO, USA) in ultrapure water (Millipore Milli-Q, 18.0 MΩ-cm). Acetaminophen, caffeine, mannitol, procaine, quinine, and methanol were all purchased from Sigma-Aldrich (St. Louis, MO, USA).
All fentanyl analogues (acetyl fentanyl, acryl fentanyl, benzyl fentanyl, butyryl fentanyl, furanyl fentanyl, methacryl fentanyl, methoxyacetyl fentanyl, phenyl fentanyl, valeryl fentanyl) were purchased as hydrochloride salts from Cayman Chemical (Ann Arbor, MI, USA) or, when possible, as 1 mg·mL−1 methanolic solutions. Powdered standards and samples were dissolved gravimetrically in methanol (Sigma-Aldrich, St. Louis, MO). The remaining opioids and narcotics (heroin, fentanyl, cocaine, U-47700) were purchased as 1 mg·mL−1 solutions in methanol or acetonitrile from Cerilliant (Round Rock, TX, USA). All sample dilutions were prepared in run buffer, and methanol was added as necessary to allow for a constant 4 % by volume organic solvent across all analyses. When increased analyte concentrations were desired, standards were concentrated to assure each sample contained a maximum of 4 % (v/v) solvent.
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2

Opioid Receptor Ligand Characterization

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U-47700 and furanyl fentanyl were procured from Cayman Chemical. Hydromorphone hydrochloride was obtained from Fagron. Buprenorphine, norBuprenorphine, fentanyl, loperamide, carfentanil, and naloxone were obtained from LGC Standards. Dulbecco's modified Eagle's medium, Opti-MEM I Reduced Serum Medium, penicillin and streptomycin (5000 IU/mL and 5000 g/mL), amphotericin B (250 g/mL), glutamine (200 mmol/L), the restriction enzyme SacI, and the DNA polymerase (Phusion polymerase) were purchased from Thermo Fisher Scientific. Primers were procured from Eurofins Genomics. [d-Ala 2 -MePhe 4 -Gly-ol]encephalin (DAMGO), fetal bovine serum (FBS), and poly-D-lysine were from Sigma Aldrich. Detailed information on chemical reagents used for processing of blood samples can be found elsewhere (13 ) .
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3

Quantification of Novel Fentanyl Analogs

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Acetylfentanyl, acrylfentanyl, furanylfentanyl and 4-fluoro-isobutyrylfentanyl were purchased as powders from Cayman Chemical (Ann Arbor, USA), diclofenac from Sigma (Stockholm, Sweden). Cryopreserved hepatocytes LiverPool (10-donor-pool), InVitro Gro HT and InVitro Gro KHB media were obtained from Bioreclamation IVT (Brussels, Belgium). Liquid chromatography mass spectrometry grade acetonitrile, water, methanol and formic acid were from Fisher Scientific (Gothenburg, Sweden), ammonium formate from Fluka (Sigma-Aldrich, Stockholm, Sweden), ammonium acetate and acetic acid from Merck (Darmstadt, Germany). The βglucuronidase/sulfatase mixture (Helix pomatia) was purchased from Roche (Mannheim, Germany). Ultra-pure water was prepared in-house using a MilliQ Gradient 10 production unit (Millipore, Billerica, USA).
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4

Sourcing and Characterization of Fentanyl Analogues

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Fentanyl, 4-ANPP, Valeryl Fentanyl, Furanyl Fentanyl, Methoxyacetyl Fentanyl, Acetyl Fentanyl, Acryl Fentanyl, and 2′-fluoro ortho-FluoroFentanyl were purchased from Cayman Chemical Company (Ann Arbor, MI). Perchloric acid was obtained from Fisher Scientific (Fair Lawn, NJ). Potassium chloride was received from Sigma-Aldrich (St. Louis, MO). Water (18.2 MΩ) was obtained using a Millipore Direct-Q ® UV water purification system (Billerica, MA).
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