Comprehensive Analysis of Drugs of Abuse in Urine Using Disposable Pipette Extraction *

  • Ellison S
  • Brewer W
  • Morgan S
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Abstract

Materials DPX-CX 1-mL tips (containing sulfonated functional groups on divinyl benzene sorbent) were obtained from DPX Labs (Columbia, SC). GC vials, inserts, and caps were purchased from Agilent (Palo Alto, CA). Acetonitrile, ethyl acetate, hex-ane, methylene chloride, isopropanol, ammonium hydroxide, and hydrochloric acid were purchased from Fisher Scientific (Pittsburgh, PA) as analytical-grade reagents. Heptafluorobu-tyric anhydride, MTBSTFA, BSTFA, and β-glucuronidase (Type H-2, from Helix pomatia) were purchased from Sigma Aldrich (St. Louis, MO). Drug-free urine was obtained from a lab vol-unteer. Previously analyzed urine case samples were provided by a local forensic toxicology laboratory. Ampoules of 6-monoacetylmorphine (6-MAM) (0.1 mg/mL), 6-MAM-d 3 (0.1 mg/mL), benzoylecgonine (BE) (0.1 mg/mL), BE-d 3 (0.1 mg/mL), morphine (1.0 mg/mL), morphine-d 3 (0.1 mg/mL), codeine (0.1 mg/mL), codeine-d 3 (0.1 mg/mL), oxy-codone (1.0 mg/mL), oxycodone-d 3 (0.1 mg/mL), am-phetamine (0.1 mg/mL), amphetamine-d 6 (0.1 mg/mL), methamphetamine (0.1 mg/mL), methamphetamine-d 5 (0.1 mg/mL), methylenedioxymethamphetamine (MDMA) (0.1 mg/mL), MDMA-d 5 (0.1 mg/mL), phencyclidine (PCP) (0.1 mg/mL), PCP-d 5 (0.1 mg/mL), 11-nor-∆ 9 -tetrahydrocannabi-nol-9-carboxylic acid (THC-COOH) (0.1 mg/mL), THC-COOH-d 3 (0.1 mg/mL), and pentobarbital-d 5 (0.1 mg/mL) were pur-chased from Cerilliant (Round Rock, TX). Quik-Check drug solutions (0.5 mg/mL) containing 20 acidic, neutral, and basic general drugs and SKF 525 A (Proadifen HCl) were purchased from Alltech (State College, PA). Working standard solutions of analytes were prepared by diluting the analytes to 10 μg/mL in methanol. Methods Spiked solutions of urine (0.2 mL) containing appropriate concentrations of analytes were prepared by adding aliquots of working standard solutions. After vortex mixing, 100 μL of 0.1 M HCl and 100 μL of acetonitrile were added and mixed. For the general drug screen, pentobarbital-d 5 and PCP-d 5 were used as internal standards for the acidic and basic drugs, re-spectively. For the targeted drugs such as BE, morphine, codeine, 6-MAM, amphetamine, and methamphetamine, deuterated standards of each analyte were used as internal standards (added prior to extraction) for reproducibility and linearity studies and external standards (added following the extraction) for recovery studies. Extraction DPX-CX 1-mL tips were used to extract drugs of abuse from the prepared urine samples. No conditioning step was per-formed prior to extraction. The wash solvent consisted of 500 μL water. The first elution solvent (for acidic/neutral drugs) consisted of 500 μL of acetonitrile; the second elution solvent (for basic drugs) consisted of 500 μL of 78:20:2 (v/v) of methy-lene chloride/isopropanol/ammonium hydroxide. The tips were loaded with the sample solution by slowly aspirating the

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Authors

  • Sparkle T Ellison

  • William E Brewer

  • Stephen L Morgan

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