Exploring Cerebrospinal Fluid: Validation of a New Method for Quantification of 39 Drugs of Abuse by LC-MS/MS

The use of cerebrospinal fluid (CSF) in post-mortem (PM) toxicological analysis is an under-addressed topic, likely due to the technical complexity of the collection of a proper sample. However, it is a matrix of significant interest since it has similar chemical and physical properties to the blood...

Full description

Saved in:
Bibliographic Details
Main Authors: Alexandra Dimitrova, Emma Beatrice Croce, Gaia Iaquinta, Elisa Ferri, Simone Faccioli, Giulia Chiavacci, Barbara Gualco, Martina Focardi, Simone Grassi, Fabio Vaiano
Format: Article
Language:English
Published: MDPI AG 2024-10-01
Series:Forensic Sciences
Subjects:
Online Access:https://www.mdpi.com/2673-6756/4/4/36
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850050173861888000
author Alexandra Dimitrova
Emma Beatrice Croce
Gaia Iaquinta
Elisa Ferri
Simone Faccioli
Giulia Chiavacci
Barbara Gualco
Martina Focardi
Simone Grassi
Fabio Vaiano
author_facet Alexandra Dimitrova
Emma Beatrice Croce
Gaia Iaquinta
Elisa Ferri
Simone Faccioli
Giulia Chiavacci
Barbara Gualco
Martina Focardi
Simone Grassi
Fabio Vaiano
author_sort Alexandra Dimitrova
collection DOAJ
description The use of cerebrospinal fluid (CSF) in post-mortem (PM) toxicological analysis is an under-addressed topic, likely due to the technical complexity of the collection of a proper sample. However, it is a matrix of significant interest since it has similar chemical and physical properties to the blood and it is less exposed to risks like PM redistribution and diffusion due to its anatomical location. This study aimed to validate a sensitive analytical method for the quantification of drugs of abuse and their metabolites (i.e., cocaine, ketamine, amphetamine, MDPV, 6-monoacetylmorphine, morphine, codeine, and methadone) through liquid chromatography–tandem mass spectrometry (LC-MS/MS). CSF was collected through ventricular puncture, and 200 µL was deproteinated with acetonitrile (600 µL). Quantification was carried out, acquiring two MRM transitions for each compound in positive ionization mode. Chromatographic separation was achieved with a C18 column. Limits of quantification ranged from 0.05 to 5 ng/mL. Bias and precision were always within the acceptance criteria. Ion enhancement and suppression effects were observed depending on the substance. The method validated here was applied to a real case, proving to be suitable for PM analysis. CSF and blood were positive for methadone (460 vs. 280 ng/mL), cocaine (125 vs. 69 ng/mL), benzoylecgonine (4640 vs. 3160 ng/mL), and lorazepam (19 vs. 25 ng/mL). In the future, this will be useful for the evaluation of CSF as a valuable alternative matrix in PM investigations.
format Article
id doaj-art-f9d55db17f314e32a79d55e69143da0b
institution DOAJ
issn 2673-6756
language English
publishDate 2024-10-01
publisher MDPI AG
record_format Article
series Forensic Sciences
spelling doaj-art-f9d55db17f314e32a79d55e69143da0b2025-08-20T02:53:33ZengMDPI AGForensic Sciences2673-67562024-10-014454655410.3390/forensicsci4040036Exploring Cerebrospinal Fluid: Validation of a New Method for Quantification of 39 Drugs of Abuse by LC-MS/MSAlexandra Dimitrova0Emma Beatrice Croce1Gaia Iaquinta2Elisa Ferri3Simone Faccioli4Giulia Chiavacci5Barbara Gualco6Martina Focardi7Simone Grassi8Fabio Vaiano9Forensic Toxicology Laboratory, Department of Health Sciences, University of Florence, 50134 Florence, ItalyForensic Toxicology Laboratory, Department of Health Sciences, University of Florence, 50134 Florence, ItalyForensic Toxicology Laboratory, Department of Health Sciences, University of Florence, 50134 Florence, ItalyForensic Medical Sciences, Department of Health Sciences, University of Florence, 50134 Florence, ItalyForensic Medical Sciences, Department of Health Sciences, University of Florence, 50134 Florence, ItalyForensic Medical Sciences, Department of Health Sciences, University of Florence, 50134 Florence, ItalyForensic Medical Sciences, Department of Health Sciences, University of Florence, 50134 Florence, ItalyForensic Medical Sciences, Department of Health Sciences, University of Florence, 50134 Florence, ItalyForensic Medical Sciences, Department of Health Sciences, University of Florence, 50134 Florence, ItalyForensic Toxicology Laboratory, Department of Health Sciences, University of Florence, 50134 Florence, ItalyThe use of cerebrospinal fluid (CSF) in post-mortem (PM) toxicological analysis is an under-addressed topic, likely due to the technical complexity of the collection of a proper sample. However, it is a matrix of significant interest since it has similar chemical and physical properties to the blood and it is less exposed to risks like PM redistribution and diffusion due to its anatomical location. This study aimed to validate a sensitive analytical method for the quantification of drugs of abuse and their metabolites (i.e., cocaine, ketamine, amphetamine, MDPV, 6-monoacetylmorphine, morphine, codeine, and methadone) through liquid chromatography–tandem mass spectrometry (LC-MS/MS). CSF was collected through ventricular puncture, and 200 µL was deproteinated with acetonitrile (600 µL). Quantification was carried out, acquiring two MRM transitions for each compound in positive ionization mode. Chromatographic separation was achieved with a C18 column. Limits of quantification ranged from 0.05 to 5 ng/mL. Bias and precision were always within the acceptance criteria. Ion enhancement and suppression effects were observed depending on the substance. The method validated here was applied to a real case, proving to be suitable for PM analysis. CSF and blood were positive for methadone (460 vs. 280 ng/mL), cocaine (125 vs. 69 ng/mL), benzoylecgonine (4640 vs. 3160 ng/mL), and lorazepam (19 vs. 25 ng/mL). In the future, this will be useful for the evaluation of CSF as a valuable alternative matrix in PM investigations.https://www.mdpi.com/2673-6756/4/4/36cerebrospinal fluiddrugs of abusepostmortem analysisLC-MS/MS
spellingShingle Alexandra Dimitrova
Emma Beatrice Croce
Gaia Iaquinta
Elisa Ferri
Simone Faccioli
Giulia Chiavacci
Barbara Gualco
Martina Focardi
Simone Grassi
Fabio Vaiano
Exploring Cerebrospinal Fluid: Validation of a New Method for Quantification of 39 Drugs of Abuse by LC-MS/MS
Forensic Sciences
cerebrospinal fluid
drugs of abuse
postmortem analysis
LC-MS/MS
title Exploring Cerebrospinal Fluid: Validation of a New Method for Quantification of 39 Drugs of Abuse by LC-MS/MS
title_full Exploring Cerebrospinal Fluid: Validation of a New Method for Quantification of 39 Drugs of Abuse by LC-MS/MS
title_fullStr Exploring Cerebrospinal Fluid: Validation of a New Method for Quantification of 39 Drugs of Abuse by LC-MS/MS
title_full_unstemmed Exploring Cerebrospinal Fluid: Validation of a New Method for Quantification of 39 Drugs of Abuse by LC-MS/MS
title_short Exploring Cerebrospinal Fluid: Validation of a New Method for Quantification of 39 Drugs of Abuse by LC-MS/MS
title_sort exploring cerebrospinal fluid validation of a new method for quantification of 39 drugs of abuse by lc ms ms
topic cerebrospinal fluid
drugs of abuse
postmortem analysis
LC-MS/MS
url https://www.mdpi.com/2673-6756/4/4/36
work_keys_str_mv AT alexandradimitrova exploringcerebrospinalfluidvalidationofanewmethodforquantificationof39drugsofabusebylcmsms
AT emmabeatricecroce exploringcerebrospinalfluidvalidationofanewmethodforquantificationof39drugsofabusebylcmsms
AT gaiaiaquinta exploringcerebrospinalfluidvalidationofanewmethodforquantificationof39drugsofabusebylcmsms
AT elisaferri exploringcerebrospinalfluidvalidationofanewmethodforquantificationof39drugsofabusebylcmsms
AT simonefaccioli exploringcerebrospinalfluidvalidationofanewmethodforquantificationof39drugsofabusebylcmsms
AT giuliachiavacci exploringcerebrospinalfluidvalidationofanewmethodforquantificationof39drugsofabusebylcmsms
AT barbaragualco exploringcerebrospinalfluidvalidationofanewmethodforquantificationof39drugsofabusebylcmsms
AT martinafocardi exploringcerebrospinalfluidvalidationofanewmethodforquantificationof39drugsofabusebylcmsms
AT simonegrassi exploringcerebrospinalfluidvalidationofanewmethodforquantificationof39drugsofabusebylcmsms
AT fabiovaiano exploringcerebrospinalfluidvalidationofanewmethodforquantificationof39drugsofabusebylcmsms