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Determination of antidepressants in whole blood using hollow-fiber liquid-phase microextraction and gas chromatography–mass spectrometry

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Abstract

A hollow-fiber liquid-phase microextraction (HF-LPME), used in three-phase mode, and combined with gas chromatography–mass spectrometry (GC–MS), was developed to quantify antidepressants and their major metabolites (amitriptyline, nortriptyline, imipramine, desipramine, clomipramine, desmethylclomipramine, fluoxetine, and norfluoxetine) in whole blood samples, using their deuterated analogs as internal standards. The HF-LPME system comprised a disposable 8-cm polypropylene porous hollow fiber, 4.0 ml of sample solution (0.5 ml of blood added to 3.5 ml of 0.1 M NaOH: donor phase), dodecane (organic phase), and 0.1 M formic acid (acceptor phase) for extraction. After stirring the system, the acceptor phase was evaporated under a nitrogen stream and resuspended in 30 μl of methanol. Derivatization was not required. A 2.0-μl aliquot of this solution was injected into a GC–MS system. The method was validated after the optimization of several parameters that may influence the extraction efficiency. The limits of quantification for all antidepressants were below the therapeutic levels (20.0 ng/ml). The average intraday and interday precisions were within 9.7 and 9.8 %, respectively, for all analytes. The calibration curves were linear in the concentration range of 20–1,200 ng/ml. The developed method was applied to seven actual postmortem samples. Tricyclic antidepressants were detected in all of the analyzed cases. To our knowledge, this is the first demonstration of usefulness of HF-LPME for analysis of antidepressants in postmortem forensic cases.

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Acknowledgments

This study was supported by the Faculty of Medicine, University of São Paulo (LIM 40/HC-FMUSP). Financial support provided by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP Grant No. 2009/08314–9) and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq Grant No. 470643/2009–9) is also gratefully acknowledged. Marcelo Filonzi dos Santos is a fellow of the FAPESP (Grant No. 2010/06530-3). Caio Caleiras Ferri is a fellow of the Programa Institucional de Bolsas de Iniciação Científica-CNPq. The authors thank the Blood Bank of Sírio Libanês Hospital for having donated blank blood samples for development of the analytical method.

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Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, University of São Paulo, Av. Professor Lineu Prestes, 580 13B, São Paulo, SP, 05508-000, Brazil

    Marcelo Filonzi dos Santos, Caio Caleiras Ferri & Mauricio Yonamine

  2. Faculty of Medicine, University of São Paulo, São Paulo, Brazil

    Saskia Carolina Seulin, Vilma Leyton & Daniel Romero Muñoz

  3. Death Verification Service, University of São Paulo, São Paulo, Brazil

    Carlos Augusto Gonçalves Pasqualucci

Authors
  1. Marcelo Filonzi dos Santos
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  2. Caio Caleiras Ferri
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  3. Saskia Carolina Seulin
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  4. Vilma Leyton
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  5. Carlos Augusto Gonçalves Pasqualucci
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  6. Daniel Romero Muñoz
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  7. Mauricio Yonamine
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Correspondence to Marcelo Filonzi dos Santos.

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dos Santos, M.F., Ferri, C.C., Seulin, S.C. et al. Determination of antidepressants in whole blood using hollow-fiber liquid-phase microextraction and gas chromatography–mass spectrometry. Forensic Toxicol 32, 214–224 (2014). https://doi.org/10.1007/s11419-014-0226-9

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  • DOI: https://doi.org/10.1007/s11419-014-0226-9

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