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7 protocols using hydromorphone

1

Quantification of Opioids and Cannabinoids in Wastewater

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Morphine, oxycodone, hydrocodone, oxymorphone, hydromorphone, Δ9-tetrahydrocannabinol (THC) and its metabolites 11-nor-9-carboxy-tetrahydrocannabinol (THCCOOH) and THCCOOH-glucuronide were purchased from Cerilliant (Round Rock, TX, USA). The deuterated analogs THC-d3, THCCOOH-d3, THCCOOH-glucuronide-d3, Morphine-d3, oxycodone-d6, hydrocodone-d6, oxymorphone-d3 and hydromorphone-d6 were also purchased from Cerilliant. Strata XC 33 µm polymeric strong cation exchange cartridges of 3 mL/60 mg for calibrators and 6 mL/200 mg for quality control (QC) and authentic wastewater samples were purchased from Phenomenex (Torrance, CA, USA). Ultra-high performance liquid chromatography (UHPLC) grade methanol, dichloromethane and ammonium hydroxide were purchased from Pharmco-Aaper (Brookfield, CT). Isopropanol, liquid chromatography mass spectrometry grade acetonitrile, and Whatman glass microfiber filters (outside diameter 4.7 cm, particle retention 1.6 μm, and thickness 0.26 mm) were from Thermo Fisher Scientific (Waltham, MA, USA).
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2

Opioid Receptor Binding Assay

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Morphine, hydromorphone, oxymorphone, butorphanol, levorphanol, nalbuphine, nalorphine, naltrexone, 6β-naltrexol, buprenorphine, pentazocine, and naloxone, were purchased from Cerilliant. Adenosine 5’-triphosphate (ATP), sodium dodecyl sulfate (SDS), dithiothreitol (DTT), dimethyl sulfoxide (DMSO), 3-(N-morpholino)propanesulfonic acid (Mops), Trizma base (Tris), isopropyl-β-D-thiogalactopyranoside (IPTG), and silica gel thin-layer chromatography (TLC) plates were from Sigma Chemical Company. Polyethyleneimine (PEI) cellulose TLC plates were from Brinkmann. Ecolume scintillation cocktail was purchased from MP Biomedical. Carrier-free sodium [35S]sulfate was from PerkinElmer. HepG2 human hepatoma cell line (ATCC HB-8065) was obtained from American Type Culture Collection. Pooled human lung S9 fraction, liver cytosol, small intestine (duodenum and jejunum) S9 fraction, and kidney S9 fraction were purchased from XenoTech, LLC. All other chemicals were of the highest grade commercially available.
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3

Certified Reference Standards for Drugs

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Certified reference standards of 6-monoacetylmorphine, alprazolam, amphetamine, buprenorphine, benzoylecgonine, carisoprodol, chlordiazepoxide, clonazepam, cocaine, codeine, cotinine, dihydrocodeine, diazepam, EDDP, fentanyl, flurazepam, hydrocodone, hydromorphone, lorazepam, MDA, MDEA, MDMA, methamphetamine, meperidine, morphine, methylphenidate, methadone, naloxone, norbuprenorphine, nicotine, nordiazepam, norfentanyl, noroxycodone, alpha-hydroxyalprazolam, oxazepam, oxycodone, oxymorphone, tapentadol, temazepam, ll-nor-9-Carboxy-Δ9-THC, and cis-tramadol were purchased, each at a concentration of 1 mg/mL, from Cerilliant Corporation (Round Rock, TX). Isotopically labeled internal standards (IS) morphine-D3, codeine- D6, hydrocodone-D3, 6-monoacetylmorphine-D6, hydromorphone-D6, oxymorphone-D3, noroxycodone-D3, and naloxone-D5 were purchased, each at a concentration of 100 μg/mL, from Cerilliant Corporation (Round Rock, TX). Drug-free urine was obtained from Bio-Rad Laboratories (Irvine, CA). High-performance liquid chromatography (HPLC) grade water, methanol, and formic acid were purchased from Fisher Scientific (Pittsburgh, PA) and ammonium formate was purchased from Sigma Aldrich (St. Louis, MO).
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4

Comprehensive Opioid Quantification Protocol

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Midazolam, Midazolam-d4 maleate, alpha-hydoxyMidazolam, alpha-hydroxyMidazolam-d4, morphine, morphine-d3, morphine-3-beta-d-glucuronide, morphine-3-beta-d-glucuronide-d3, morphine-6-beta-d-glucuronide, morphine-6-beta-d-glucuronide-d3, hydromorphone, hydromorphone-d6, fentanyl, fentanyl-d5, norfentanyl, and norfentanyl-d5-oxalate were all purchased from Cerilliant (Round Rock, Texas). Dexmedetomidine HCl was purchased from Athenex Pharmaceuticals (Buffalo, New York) and dexmedetomidine-d4-L tartrate was purchased from Toronto Research Chemical (Toronto, Ontario). LC-MS grade reagents; methanol, acetonitrile, formic acid and water were purchased from Fisher Scientific (Pittsburgh, PA, USA). Six lots of human plasma was purchased from Lampire Biological Laboratories (Pipersville, Pa).
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5

Quantification of Opioid Analytes in Serum

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Two independent lot numbers of certified reference standards of morphine, morphine-3-β-glucuronide, morphine-6-β-glucuronide, normorphine, hydromorphone as well as a single lot of the isotopically labeled internal standards (IS) morphine-d3 morphine-3-β-glucuronide-d3, morphine-6-β-glucuronide-d3, and hydromorphone-d6 were all purchased from Cerilliant Corporation (Round Rock, TX). Interference standards (see Supplemental Table 1) were obtained from Cerilliant Corporation. Drug-free serum was obtained from Bio-Rad Laboratories (Irvine, CA). High-performance liquid chromatography (HPLC) grade water and methanol were purchased from Fischer Scientific (Pittsburgh, PA) and formic acid was acquired from Sigma Aldrich (St. Louis, MO). For selectivity, matrix effects and endogenous studies, remnant human serum was acquired from vacutainer serum separator tubes (SST) (BD, Franklin Lakes, NJ) and plasma for the plasma to serum comparison from lithium heparin tubes (BD, Franklin Lakes, NJ) via an Institutional Review Board (IRB)-approved protocol through The Johns Hopkins University School of Medicine.
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6

Comprehensive Analytical Reference Standards

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Certified reference materials of amitriptyline, bupropion, citalopram, desipramine, desmethylcitalopram, duloxetine, fluoxetine, imipramine, mirtazapine, nortriptyline, paroxetine, sertraline and trazodone were purchased from LGC Standards (Teddington, London, UK); certified reference materials of clomipramine, desmethylvenlafaxine, doxepin, hydroxybupropion, norfluoxetine, norsertraline, trimipramine, venlafaxine, 7-aminoclonazepam, 7-aminoflunitrazepam, alprazolam, bromazepam, clonazepam, diazepam, flunitrazepam, lorazepam, midazolam, nitrazepam, nordiazepam, oxazepam, temazepam, zolpidem, 6-monoacetylmorphine, N-desmethyltramadol, buprenorphine, codeine, fentanyl, hydrocodone, hydromorphone, meperidine, methadone, morphine, naloxone, naltrexone, oxycodone, oxymorphone, tramadol, bupropion-d 9 , citalopram-d 6 , clonazepam-d 4 , codeine-d 3 , diazepam-d 5 , duloxetine-d 3 and morphine-d 3 from Cerilliant (Round Rock, TX, USA); methanol, acetonitrile, ammonium formate and formic acid from Merck (Darmstadt, Germany); ultrapure deionized water was purified by Milli-Q from Millipore (Billerica, MA, USA). All solvents used in the extraction procedure were HPLC grade. Quantisal™ oral fluid collection devices and elution buffer were purchased from Immunalysis (Pomona, CA, USA).
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7

Quantification of Opioid Compounds

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Methanolic solutions of morphine, M3G, morphine-6-glucuronide (M6G), normorphine, hydromorphone and 6monoacetylmorphine (MAM) (1 mg/mL) and the deuterated internal standards (IS) morphine-d3, M3G-d3, M6G-d3 and MAM-d3 (0.1 mg/mL) were obtained from Cerilliant (delivered by Sigma-Aldrich, Buchs, Switzerland). Water was purified with a Purelab Ultra millipore filtration unit (Labtech, Villmergen, Switzerland) and acetonitrile of HPLC grade was obtained from Fluka (Buchs, Switzerland). All other chemicals used were from Merck (Zug, Switzerland) and of the highest grade available.
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