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. 2023 Jan;41(1):94-104. doi: 10.1007/s11419-022-00640-y. Epub 2022 Sep 13.

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.

Keywords: Cerumen; Liquid–liquid extraction; Postmortem samples; Psychoactive substances; Ultra-performance liquid chromatography–tandem mass spectrometry.

MeSH terms

  • Cerumen* / chemistry
  • Chromatography, Liquid / methods
  • Psychotropic Drugs / analysis
  • Reproducibility of Results
  • Solvents
  • Tandem Mass Spectrometry* / methods

Substances

  • Psychotropic Drugs
  • Solvents