Optimized and Validated DBS/MAE/LC–MS Method for Rapid Determination of Date-Rape Drugs and Cocaine in Human Blood Samples—A New Tool in Forensic Analysis

Paweł Stelmaszczyk,Renata Wietecha-Posłuszny,Ewa Gacek

doi:10.3390/separations8120249

Paweł Stelmaszczyk, Renata Wietecha-Posłuszny + Show 1 more

Open Access

https://doi.org/10.3390/separations8120249

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Journal: Separations	Publication Date: Dec 18, 2021
Citations: 8	License type: CC BY 4.0

Affiliation: Jagiellonian University

Abstract

The aim of this work was to develop a new method for the determination of selected substances from the date-rape drugs group: ketamine, benzodiazepines and cocaine. The method is based on the dried blood spot method which seems to be a suitable tool in the analysis of tested substances. The extraction process based on microwave-assisted extraction was optimized to enable optimal conditions for the isolation of a wide range of analytes from blood samples collected on DBS cards. The extraction with ethyl acetate with a buffer of pH = 9 carried out at a temperature of 50 °C for 15 min ensured high extraction efficiency of the tested analytes. The optimized method was validated. Limits of detection (LOD = 4.38–21.1 ng/mL) and quantification (LOQ = 14.6–70.4 ng/mL), inter- and intra-day precision (CV = 1.37–13.4% and 3.39–14.8%, respectively), recovery (RE = 93.0–112.4%) and matrix effect (ME = 98.4–101.6%) were determined. The validation results indicate the possibility of using the proposed method in the analysis of real blood samples collected from victims of sexual assault.

Highlights

Drug-facilitated sexual assault (DFSA) has been a major area of forensic work in recent years
The developed and validated dried blood spot (DBS)/microwave assisted extraction (MAE)/liquid chromatography with mass spectrometry (LC–MS) method presented in this work could be used for the obtainment of blood samples and their qualitative and quantitative analysis
The conditions of MAE extraction were optimized during this study, with the optimal parameters chosen based on the experimental design

Summary

Introduction

Drug-facilitated sexual assault (DFSA) has been a major area of forensic work in recent years. DRDs usually cause helplessness and loss of consciousness, which in turn leads to the victim’s inability to repel the attacker’s assault. These substances are generally tasteless, colorless, odorless and dissolve well in water and alcohol [2]. Benzodiazepines, flunitrazepam and diazepam, ketamine, γ-hydroxybutanoic acid (GHB), cocaine and antidepressants are the most commonly used chemicals in sexual assaults. Many of these drugs may cause depression of the central nervous system (CNS), for example, benzodiazepines, which cause somnolence, sedation and muscle relaxation, and ketamine, which at high doses causes hallucinations and withdrawal from reality [3]. A small part of a given substance remains unchanged in the blood and the resulting metabolites are quickly eliminated from the body through urine [4]

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