Detection of Morphine and Opioids in Fingernails: Immunohistochemical Analysis and Confirmation with Ultra-High-Performance Liquid Chromatography Coupled with High-Resolution Mass Spectrometry

Abstract

This study aimed to investigate the detection of morphine in fingernails from forensic autopsies using immunohistochemistry (IHC), with confirmation by ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS). A primary antibody specific to morphine and a secondary antibody conjugated to horseradish peroxidase (HRP) was used. IHC on specimens of Subjects A and B (both drug addicts) resulted in the detection of morphine on a cell layer of the nail plate matrix. UHPLC-HRMS and GC-MS analysis showed that Subject A had a morphine concentration of 0.35 ng/mg in the fingernail and 472 ng/mL in the blood, while Subject B reached 1.23 ng/mg in the fingernail and 360 ng/ml in the blood. Most of those matrices were positive for codeine, methadone, EDDP, and 6-MAM. The use of IHC in Subject C (a former addict) showed no positivity for morphine in the fingernail, while the UHPLC-HRMS analysis confirmed its absence in the fingernail and blood. Additionally, an analysis of the scalp or pubic hair of the subjects was carried out using UHPLC-HRMS. The results suggest that IHC can be used to establish the site of accumulation of morphine in the nail matrix; for postmortem diagnosis; and that basic substances can be detected by UHPLC-HRMS. There are no previous studies on the use of IHC as a technique for forensic purposes in unconventional matrices, such as nails.

Detection of Three Opioids (Morphine, Codeine and Methadone) and Their Metabolites (6-Monoacetylmorphine and 2-Ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine) in Larvae of Lucilia sericata Species by UHPLC-TF-MS and Validation

“…However, the analysis on unconventional matrices is very important in cases where the remains are in an advanced state of decomposition; thus, conventional matrices are no more available. Different studies are already present in the literature for the detection of opioids in alternative matrices [ 22 , 23 ]. In unconventional matrices such as larvae, quantitative data could be even more difficult in relation to the mechanism of death.…”

Machine learning-enhanced drug testing for simultaneous morphine and methadone detection in urinary biofluids

“…Over the past decades, various methodologies have emerged to detect MOR, MET, and UA accurately. These encompass high-performance liquid chromatography (HPLC) 14 , liquid chromatography-mass spectrometry (LC–MS) 15 , spectroscopy 16 , supercritical fluid chromatography 17 , and chemiluminescence 18 . Although these techniques demonstrate commendable sensitivity and remarkable reliability, their practical utility is constrained by certain drawbacks, including time-consuming procedures, the necessity for sample pre-treatment, analytical complexity, and the associated costs and labor intensiveness 19 , 20 .…”

The Identification of Drug Abuse

“…Scendoni and Coll. first report results for morphine determination in an unusual post-mortem matrix: in fingernails [ 12 ]. Data were first acquired by immunohistochemistry and subsequently confirmed by ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry.…”