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Development and validation of a sensitive LC–MS/MS method to analyze NBOMes in dried blood spots: evaluation of long-term stability

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Abstract

Purpose

We evaluated the use of dried blood spots (DBS) to determine seven NBOMes by liquid chromatography–tandem mass spectrometry (LC–MS/MS), and also evaluated the stability of these compounds in this dried matrix.

Methods

An LC–MS/MS method was developed and fully validated to quantify seven NBOMes (25C-, 25H-, 25I-, 25B-, 25G-, 25D- and 25E-NBOMe) in DBS samples. The extraction procedure was optimized using mixture design experiment. Stability study was performed in two different concentrations over 180 days at three different storage temperatures.

Results

Good linearity, and limits of detection and quantitation of 0.05 and 0.1 ng/mL, respectively, were obtained. The interday imprecision (n = 15) and bias (n = 15) were not higher than 11.4 and 10.3%, respectively, and no carryover was observed. All analytes remained stable in DBS at − 20, 4 °C and even at room temperatures for 180 days, except 25B-NBOMe and 25I-NBOMe which experienced degradation (22 and 21%, respectively) of the initial concentration at room temperature after 180 days of study. The method was applied to a DBS of an authentic postmortem blood from an NBOMe user, and it was found to be reliable with good selectivity and specificity.

Conclusions

DBS has been found to allow reliable, sensitive, accurate and robust detection and quantification of seven NBOMes via LC–MS/MS. Also, DBS provided great stability to most of the compounds at room temperature, and no degradation was observed for DBS kept at 4 and − 20 °C. This is the first trial to analyze NBOMes in DBS samples to our knowledge.

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Acknowledgements

The authors thank Fundação de Amparo à Pesquisa do Estado de São Paulo – FAPESP (Process Number 2015/10650-8), Conselho Nacional de Desenvolvimento Científico e Tecnológico – CNPq (Process Number 830525/1999-8), Fundo de Apoio ao Ensino, à Pesquisa e à Extensão – FAEPEX (Agreement 519.292 MF 86739) and Superintendência da Polícia Técnico-Científica.

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Authors and Affiliations

  1. Campinas Poison Control Center, Faculty of Medical Sciences, University of Campinas, Campinas, São Paulo, 13083-859, Brazil

    Kelly Francisco da Cunha & Jose Luiz Costa

  2. ThoMSon Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas, Campinas, São Paulo, 13083-970, Brazil

    Marcos Nogueira Eberlin

  3. Faculty of Pharmaceutical Sciences, University of Campinas, Campinas, São Paulo, 13083-859, Brazil

    Jose Luiz Costa

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  1. Kelly Francisco da Cunha
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Correspondence to Jose Luiz Costa.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee (Comitê de Ética em Pesquisa da UNICAMP – CEP, CAAE 58452716.2.0000.5404 and Superintendência da Polícia Técnico-Científica, ofício No. 766/2015/ATS/SPTC-SSP) and with the 1964 Helsinki Declaration.

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da Cunha, K.F., Eberlin, M.N. & Costa, J.L. Development and validation of a sensitive LC–MS/MS method to analyze NBOMes in dried blood spots: evaluation of long-term stability. Forensic Toxicol 36, 113–121 (2018). https://doi.org/10.1007/s11419-017-0391-8

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