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

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Abstract

This article describes an easy and innovative extraction procedure for cocaine and its primary metabolite, benzoylecgonine (BE), from hair consisting of sonication with H2O/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%.

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Authors and Affiliations

  1. Institute of Legal Medicine, Faculty of Medicine, University of Santiago de Compostela, San Francisco, s/n, 15782, Santiago de Compostela, Spain

    P. López, A. M. Bermejo & M. J. Tabernero

  2. Institute of Legal Medicine, Faculty of Medicine, Catholic University of the Sacred Heart, Largo Francesco Vito 1, 00168, Rome, Italy

    S. Martello, Eleonora De Vincenzi & M. Chiarotti

Authors
  1. P. López
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  2. S. Martello
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  3. A. M. Bermejo
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  4. Eleonora De Vincenzi
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  5. M. J. Tabernero
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  6. M. Chiarotti
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Correspondence to S. Martello.

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López, P., Martello, S., Bermejo, A.M. et al. Validation of ELISA screening and LC–MS/MS confirmation methods for cocaine in hair after simple extraction. Anal Bioanal Chem 397, 1539–1548 (2010). https://doi.org/10.1007/s00216-010-3684-x

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  • DOI: https://doi.org/10.1007/s00216-010-3684-x

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