Validation of ELISA screening and LC-MS/MS confirmation methods for cocaine in hair after simple extraction

Anal Bioanal Chem. 2010 Jun;397(4):1539-48. doi: 10.1007/s00216-010-3684-x. Epub 2010 Apr 17.

Abstract

This article describes an easy and innovative extraction procedure for cocaine and its primary metabolite, benzoylecgonine (BE), from hair consisting of sonication with H(2)O/0.1% formic acid for 4 h. The same extract was used for screening with an enzyme-linked immunoassay (ELISA) and confirmation by liquid chromatography-tandem mass spectrometry (LC-MS/MS). For the ELISA screening test a cutoff of 0.5 ng/mg was used according to the Society of Hair Testing recommendations. LC-MS/MS limits of detection (LODs) were established to be 10 pg/mg and 1 pg/mg for cocaine and BE, respectively. Linearity was obtained over a range of 0.2-5 ng/mg for BE (target analyte) in the ELISA screening test, while in the LC-MS/MS method the range was 0.10-10 ng/mg for cocaine and 0.01-10 ng/mg for BE. Intra- and interbatch coefficients of variation and mean relative errors were less than 20% for all analytes and concentrations studied. The validated ELISA and LC-MS/MS methods were applied to 48 hair samples and the results of both methods were compared; ELISA demonstrated a sensitivity and specificity of 89.2% and 10.8%.

Publication types

  • Validation Study

MeSH terms

  • Adult
  • Chromatography, Liquid
  • Cocaine / analogs & derivatives
  • Cocaine / analysis*
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Hair / chemistry*
  • Humans
  • Male
  • Sensitivity and Specificity
  • Tandem Mass Spectrometry
  • Young Adult

Substances

  • Cocaine