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Development and validation of an ultra-performance liquid chromatography–tandem mass spectrometric method for the determination of 25 psychoactive drugs in cerumen and its application to real postmortem samples

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Abstract

Purpose

In the present study, a method for the detection of 25 psychoactive substances in cerumen was developed and validated. This method targets opiates, cocaine, antidepressants, benzodiazepines, antipsychotics and antiparkinsons.

Methods

Analysis was performed on a SCIEX Triple Quad 6500+ system after liquid–liquid extraction. Methanol with 1% acetic acid was chosen as the extraction solvent. After the addition of the solvent, samples were vortexed, sonicated, centrifuged and directly injected into the liquid chromatography–tandem mass spectrometry system.

Results

The method was found to be selective and sensitive (limit of detection: 0.017 ng–0.33 ng/mg), the assay was linear for all analytes with linear regression coefficient ranging 0.9911–1.00. The values for intra-assay precision was between 4.34 and 14.6% and inter-assay precision between 5.81 and 17.7%, with accuracy within the acceptable criteria for all analytes. All analytes in cerumen specimens were stable for 48 h at 4 °C and 72 h at − 20 °C, whilst no significant matrix effect or carryover was observed. Applicability was proven by analyzing cerumen samples from 25 deceased with a history of drug abuse. All analytes were detected in real samples, thus confirming the sensitivity of the developed method.

Conclusions

According to our knowledge, it is the first time that a method for the simultaneous detection of 25 psychoactive drugs in cerumen was developed, fully validated and finally applied to 25 postmortem samples.

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Acknowledgements

The authors wish to acknowledge the technical employees of the Forensic Service of Thessaloniki for providing the biological samples.

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Authors and Affiliations

  1. Toxicology Laboratory, Forensic Service of Thessaloniki, Ministry of Justice, Thessaloniki, Greece

    Orthodoxia Mastrogianni & Heleni Zagelidou

  2. Laboratory of Forensic Medicine and Toxicology, Medical School, Aristotle University of Thessaloniki, Thessaloniki, Greece

    Orthodoxia Mastrogianni, Amvrosios Orfanidis, Evdokia Brousa, Dimitrios-Phaedon Kevrekidis & Nikolaos Raikos

Authors
  1. Orthodoxia Mastrogianni
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  2. Amvrosios Orfanidis
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  3. Evdokia Brousa
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  5. Heleni Zagelidou
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  6. Nikolaos Raikos
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Correspondence to Orthodoxia Mastrogianni.

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The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in this study were in accordance with the ethical standards of the national committee and with the 1964 Declaration of Helsinki. Biological fluids (blood, urine) and cerumen collection from decedents was made by judicial authorities, and the samples were sent to our institute for toxicological analysis at their request. This article does not contain any studies with living human participants or animals performed by any of the authors.

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Mastrogianni, O., Orfanidis, A., Brousa, E. et al. Development and validation of an ultra-performance liquid chromatography–tandem mass spectrometric method for the determination of 25 psychoactive drugs in cerumen and its application to real postmortem samples. Forensic Toxicol 41, 94–104 (2023). https://doi.org/10.1007/s11419-022-00640-y

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