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9 protocols using codeine

1

Extraction and Analysis of Opioids

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Meconin-d3 and meconin were purchased from Cayman Chemical (Ann Arbor, MI) as solids and codeine, codeine-d3, morphine, morphine-d3, 6-monoacetylmorphine-d3 (MAM-d3), and 6MAM were purchased from Cerilliant (Round Rock, TX) as 1 mg/mL ampoules. All reagents (ACS grade) and all solvents (HPLC grade) were purchased from Thermo-Fisher Scientific (Hanover Park, IL). Solid-phase extraction (SPE) cartridges (ZSDAU020, 200-mg bed, 10 mL reservoir) were purchased from United Chemical Technologies (Bristol, PA). Extracts were evaporated in a TurboVap LV II purchased from Biotage (Charlotte, NC) under a stream of nitrogen at 60°C. Umbilical cord tissue aliquots were homogenized in a Next Advance Bullet Blender (Averill Park, NY) using 3 stainless steel wood screws.
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2

Quantification of Illicit Drugs in Cough Syrups

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Carbinoxamine, chlorpheniramine, and chlorpheniramine-d6 were purchased from TRC (North York, Canada). Individual solutions of 6-AM, 7-aminonitrazepam, 7-aminonimetazepam, amphetamine, benzoylecgonine, C6G, codeine, codeine-d6, ketamine, methamphetamine, mephedrone, 4-methylephedrine, 3,4-methylenedioxymethamphetamine hydrochloride, M3G, M6G, morphine, morphine-d6, and norketamine (each with a concentration of 1 mg mL−1) were purchased from Cerilliant (Round Rock, TX, USA). Three brands of cold syrups (Consrine, ssBuron, and Pabron) manufactured in Taiwan were purchased from local pharmacies. Consrine contains codeine phosphate (0.48 mg mL−1), chlorpheniramine (0.25 mg mL−1), dl-methyl ephedrine HCl (1.6 mg mL−1), and glycyrrhizic acid (0.25 mg mL−1), ssBuron contains codeine phosphate (0.5 mg mL−1), guaifenesin (3.33 mg mL−1), chlorpheniramine maleate (0.14 mg mL−1), and caffeine (1.33 mg mL−1), Pabron contains codeine phosphate (0.48 mg mL−1), dl-methyl ephedrine HCl (2.5 mg mL−1), and carbinoxamine maleate (0.4 mg mL−1). Formic acid and methanol (high-performance LC grade) (≥ 98%) were purchased from J.T. Baker (Pittsburgh, PA, USA). Ultrapure water (18.2 mΩ·cm) obtained from a Milli-Q ultrapure water system (Millipore, Billerica, MA, USA) was used in this study.
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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

Analytical Characterization of AAIDs

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AAID standards and all the reagents used in the study were of analytical grade. Diclofenac, ibuprofen, indomethacin, ketoprofen, naproxen, and paracetamol were purchased from Fluka (Switzerland). Acetylsalicylic acid, mefenamic acid, and phenylbutazone were obtained from Sigma (Switzerland), while codeine was purchased from Cerilliant (TX, USA). Physical and chemical properties of AAIDs are given in Table S1. HCl and NaOH were supplied from Merck Co (Darmstadt, Germany). A nylon filter with a 0.45 μm pore diameter was obtained from Sartorius (Göttingen, Germany). Deionized water was obtained from a Millipore Milli-Q Plus water purifier (Merck, MA, USA). The nitrogen gas for high performance liquid chromatography-mass spectrometry (HPLC–MS) was acquired from a nitrogen generator (Peak Scientific, Scotland, UK).
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5

Quantitative Analysis of Illicit Drugs and Novel Psychoactive Substances

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LC–MS-grade methanol (purity, ≥99.9%) was purchased from Merk (Darmstadt, Germany). HPLC-grade acetonitrile (purity, ≥99.9%) was purchased from J.T. Baker (Phillipsburg, NJ, USA). Ten standards of illicit drugs, methamphetamine (1 mg/mL), amphetamine (1 mg/mL), ketamine (1 mg/mL), norketamine (1 mg/mL), morphine (0.1 mg/mL), codeine (1 mg/mL), Tetrahydrocannabinol (THC, 1 mg/mL), 11-nor-9-carboxy-THC (THC-COOH, 0.1 mg/mL), 3,4-methylenedioxymethamphetamine (MDMA, 1 mg/mL), mephedrone (1 mg/mL), flunitrazepam (1 mg/mL), 7-aminoflunitrazepam (1 mg/mL), nimetazepam (1 mg/mL), 7-aminonimetazepam (1 mg/mL), and zopiclone (1 mg/mL) were purchased from Cerilliant (Round Rock, TX, USA). Three standards of NPSs, 4′-chloro-alpha-pyrrolidinopropiophenone (4-Cl-α-PPP, 1 mg, purity, ≥98%), 4′-Fluoro-α-pyrrolidinopentiophenone (4-F-α-PVP, 1 mg, purity, ≥98%), and 1-pentyl-3-(2-bromophenylacetyl)indole (JWH-249, 1 mg, purity, ≥98%) were purchased from Cayman Chemicals (Ann Arbor, MI, USA). Sodium azide (500 mg, purity, ≥99%) was purchased from Sigma-Aldrich (St. Louis, MO, USA).
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6

Analytical Standards for Pharmaceutical Analysis

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AAID standards, and all chemicals were of analytical grade. While codeine was purchased from Cerilliant (TX, USA), acetylsalicylic acid, mefenamic acid and phenylbutazone were obtained from Sigma (Switzerland). Acetaminophen, diclofenac, ibuprofen, indomethacin, ketoprofen, and naproxen were purchased from Fluka (Switzerland). HCl, NaOH, FeCl2, FeCl3, NH3, HNO3, HCOOH, NH4OH, NH4HCO2 and C2H3NaO2 were also obtained from Merck Co (Darmstadt, Germany). A nylon filter with a 0.45 μm pore diameter was obtained from Sartorius (Göttingen, Germany). Deionized water was obtained from a Millipore Milli-Q Plus water purifier (Merck, MA, USA). The nitrogen gas for high performance liquid chromatography-mass spectrometry (HPLC–MS) was acquired from a nitrogen generator (Peak Scientific, Scotland, UK).
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7

Analytical Standards for Opioid Quantification

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The analytical reference standards for codeine, C6G, norcodeine, morphine, M3G, M6G and the internal standards d6-morphine, d6-codeine and d3-M6G were obtained from Cerilliant Corporation, TX, USA. High performance liquid chromatography grade or better solvents were purchased from the following manufacturers: acetonitrile (ACN) and water from Burdick & Jackson, MI, USA; methanol and buffer reagents from Thermo Fisher Scientific, NJ, USA; and formic Acid, 98%, from Sigma-Aldrich, MO, USA.
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8

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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9

Deuterated Opioid Extraction from Plasma

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Codeine, hydrocodone and their deuterated analogues were purchased from Cerilliant (Round Rock, TX, USA), and used without further purification. HPLC grade acetonitrile and methanol were bought from Caledon Laboratory Chemicals (Georgetown, ON, Canada). Distilled deionized water (18 MΩ) was produced in-house using a Millipore (Billerica, MA, USA) Integral 10 water purification system. Plasma samples were purchased from Bioreclamation IVT (Baltimore, MD, USA). Target analytes and internal standards (deuterated analogues) were spiked in the plasma (v/v) the day before extraction. Bio-SPME mixed mode probes (i.e. C18-SCX particles, 45 μm thickness, 15 mm coating length) were kindly provided by Millipore-Sigma (Supelco; Bellefonte, PA, USA).
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