Gas Chromatography Mass Spectrometry (GC‐MS) for Identification of Designer Stimulants Including 2C Amines, NBOMe Compounds, and Cathinones in Urine

Hemamalini Ketha1, Milad Webb1, Larry Clayton1, Sean Li1

1 Department of Pathology, University of Michigan Health System, Ann Arbor
Publication Name:  Current Protocols in Toxicology
Unit Number:  Unit 4.43
DOI:  10.1002/cptx.33
Online Posting Date:  November, 2017
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Abstract

Phenethylamine derivatives are being increasingly exploited for recreational use as “designer” stimulants designed to mimic psychostimulant properties of amphetamine or other illicit substances like 3,4‐methylenedioxymethamphetamine (MDMA [ecstasy]). Clandestine operations meticulously design phenethylamines so the user can bypass legal action when detected, as many of these are yet to be regulated by government authorities. Substituted phenethylamines or 2C amines, N‐methoxybenzyl derivatives of the corresponding 2C amines commonly known as NBOMe compounds, and cathinones are among the most commonly abused phenethylamines. Current FDA‐approved assays used in screening for illicit drug use lack the sensitivity needed to detect designer stimulants making it challenging for toxicologists to accurately identify these compounds. Gas chromatography mass spectrometry (GC‐MS) is a sensitive method for identifying designer stimulants. This unit describes and compares two qualitative GC‐MS methods for identifying 2C amines, NBOMe compounds, and cathinones in urine. © 2017 by John Wiley & Sons, Inc.

Keywords: designer amines; GC‐MS; phenethylamines; designer stimulants; designer amphetamines

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Identifying Designer Stimulants in Urine Using Organic Extraction Followed by GC‐MS Analysis
  • Basic Protocol 2: Identifying Designer Stimulants in Urine Using Organic Extraction Followed by Derivatization and GC‐MS Analysis
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Identifying Designer Stimulants in Urine Using Organic Extraction Followed by GC‐MS Analysis

  Materials
  • Urine sample
  • Extraction buffer (see recipe)
  • Extraction solvent (see recipe)
  • 100 µg/ml dioctylphthalate (see recipe)
  • Ethyl acetate, LC‐MS grade (e.g., Sigma‐Aldrich, cat. no. 34972)
  • GC‐MS quality control solution (see recipe)
  • 20 × 125 mm screw‐top glass tube
  • Laboratory rocker
  • Centrifuge
  • 13 × 100 mm test tubes
  • Nitrogen evaporator and evaporation tubes
  • Vortex
  • 250‐µl plastic screw‐cap vials
  • Autosampler
  • GC‐MS system (e.g., Agilent GC‐7890B and MS‐5977A) with capillary and mass ionization detector
  • Additional reagents and equipment for GC‐MS (see Agilent Operation Manual available at https://www.agilent.com/cs/library/usermanuals/Public/G3430‐90011.pdf)

Basic Protocol 2: Identifying Designer Stimulants in Urine Using Organic Extraction Followed by Derivatization and GC‐MS Analysis

  Materials
  • Urine sample
  • 5.0 N aqueous NaOH
  • Hexane (e.g., Sigma‐Aldrich, cat. no. 650420)
  • Pentaflouropropionic acid anhydride (PFPA; e.g., Sigma‐Aldrich, cat. no. 245917)
  • Pyridine, anhydrous (e.g., Sigma‐Aldrich, cat. no. 270970)
  • Acetic anhydride (e.g., Sigma‐Aldrich, cat. no. 45830)
  • Ethyl acetate, LC‐MS grade (e.g., Sigma‐Aldrich, cat. no. 34972)
  • GC‐MS quality control solution (see recipe)
  • 13 × 100 mm glass test tubes
  • Vortex
  • Centrifuge
  • 50°C water bath
  • Nitrogen evaporator
  • Screw‐cap vials
  • Autosampler
  • GC‐MS system (e.g., Agilent GC‐7890B and MS‐5977A) with capillary and mass ionization detector
  • Additional reagents and equipment for GC‐MS (see Agilent Operation Manual available at https://www.agilent.com/cs/library/usermanuals/Public/G3430‐90011.pdf)
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Figures

Videos

Literature Cited

Literature Cited
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  Kanamori, T., Inoue, H., Iwata, Y., Ohmae, Y., & Kishi, T. (2002). In vivo metabolism of 4‐bromo‐2,5‐dimethoxyphenethylamine (2C‐B) in the rat: Identification of urinary metabolites. Journal of Analytical Toxicology, 26, 61–66. doi: 10.1093/jat/26.2.61.
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  Moeller, K. E., Lee, K. C., & Kissack, J. C. (2008). Urine drug screening: Practical guide for clinicians. Mayo Clinic Proceedings, 83, 66‐76. doi: 10.4065/83.1.66.
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  Weaver, M. F., Hopper, J. A., & Gunderson, E. W. (2015). Designer drugs 2015: Assessment and management. Addiction Science & Clinical Practice, 10, 8. doi: 10.1186/s13722‐015‐0024‐7.
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