Minkoff Huestis1*, Buckley Greene

ABSTRACT

Solid-Phase Extraction (SPE) remains the traditional method applied to postmortem blood, offering strong purification capability but presenting notable drawbacks, including operator dependence, limited batch capacity, and relatively high operational costs. Accelerated Solvent Extraction (ASE), an automated extraction technique, has emerged as a promising alternative capable of reducing manual variability, increasing sample throughput, and shortening overall processing times. This study aimed to validate an ASE-based workflow for whole-blood toxicological analysis and assess its extraction performance in comparison with conventional SPE. Whole-blood specimens obtained from cadavers undergoing medico-legal autopsy at the University of Milan were fortified with a panel of psychoactive drugs, antagonists, therapeutic agents, and anesthetics. Each sample was subjected to both ASE and SPE extraction protocols, followed by instrumental analysis using a Thermo Scientific™ TSQ Fortis™ II Triple-Quadrupole Mass Spectrometer. Comprehensive method development and validation procedures were conducted to evaluate accuracy, precision, recovery, matrix effects, and overall suitability of ASE for forensic applications. Comparative assessment of the extractive approaches demonstrated the feasibility and analytical robustness of the ASE technique, supporting its potential as an efficient and operator-independent alternative to traditional SPE in forensic toxicology laboratories. Keywords: Forensic Toxicology; Accelerated Solvent Extraction (ASE); SolidPhase Extraction (SPE); Biological Matrices; riple-Quadrupole Mass Spectrometry; Toxicological Analysis. INTRODUCTION Over the years, numerous extraction approaches have been introduced with the goal of improving efficiency, speeding up sample preparation, and reducing the amount of solvent required. Among these, Solid-Phase Extraction (SPE) continues to be widely used because it offers clear benefits over classical techniques such as liquid–liquid extraction [1]. SPE reduces solvent consumption, minimizes manual handling, shortens overall processing time, and provides better selectivity, which is why it remains a preferred method in several analytical fields, including forensic toxicology [2,3]. SPE has long been established as a dependable samplepreparation method and forms a key component in toxicological investigations. The technique is based on principles similar to partition chromatography, and ov

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