Sensitive method for detection of cocaine and associated analytes by liquid chromatography-tandem mass spectrometry in urine

J Anal Toxicol. 2009 Oct;33(8):447-55. doi: 10.1093/jat/33.8.447.

Abstract

Cocaine (COC) is a potent CNS stimulant that is metabolized to benzoylecgonine (BE) and further metabolized to minor metabolites such as m-hydroxybenzoylecgonine (m-HOBE). COC is also metabolized to norcocaine (NC). Cocaethylene (CE) is formed when cocaine and ethyl alcohol are used simultaneously. Anhydroecgonine methyl ester (AEME) is a unique marker following smoked cocaine, and anhydroecgonine ethyl ester (AEEE) is found in cocaine smokers who also use ethyl alcohol. We developed a liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for the detection and quantitation of COC, BE, NC, CE, m-HOBE, AEME, and AEEE in urine. Two hundred samples previously analyzed by gas chromatography (GC) coupled with MS were extracted using solid-phase extraction. Chromatographic separation was achieved using a gradient consisting of mobile phase A [20 mM ammonium formate (pH 2.7)] and mobile phase B (methanol/acetonitrile, 50:50), an XDB-C(8) (50 x 2.1 mm, 1.8 microm) column and a flow rate of 270 microL/min. Concentrations were calculated by comparing the peak-area with the internal standard and plotted against a standard curve. The assay displayed linearity from 1.0 to 100 ng/mL. Within- and between-run coefficients of variation were < 10% throughout the linear range. A method comparison between GC-MS and LC-MS-MS showed good correlation for COC (r(2) = 0.982) and BE (r(2) = 0.955). We report here on a sensitive method to identify clinically and forensically relevant cocaine and associated analytes at concentrations as low as 1.0 ng/mL.

MeSH terms

  • Chromatography, High Pressure Liquid
  • Cocaine / analogs & derivatives
  • Cocaine / urine*
  • Cocaine-Related Disorders / diagnosis
  • Cocaine-Related Disorders / urine*
  • Dopamine Uptake Inhibitors / urine*
  • Ethanol / blood
  • Gas Chromatography-Mass Spectrometry
  • Humans
  • Reproducibility of Results
  • Solid Phase Extraction
  • Spectrometry, Mass, Electrospray Ionization
  • Substance Abuse Detection / methods*
  • Tandem Mass Spectrometry

Substances

  • Dopamine Uptake Inhibitors
  • 3-hydroxybenzoylecgonine
  • Ethanol
  • norcocaine
  • anhydroecgonine methyl ester
  • benzoylecgonine
  • Cocaine
  • ethyl ecgonine