First-Line Toxicological Screening with Fully Automated Extraction

Area of Science:

• Clinical Chemistry
• Analytical Chemistry
• Forensic Toxicology

Background:

• Clinical toxicology laboratories require rapid, unambiguous methods for identifying compounds in overdose or poisoning cases.
• Existing methods may lack the speed and comprehensive identification needed for timely clinical decisions.
• The demand for high-throughput screening in toxicology necessitates advanced analytical techniques.

Purpose of the Study:

• To develop and validate a fully automated screening procedure for identifying and quantifying 245 clinically relevant compounds.
• To achieve unambiguous compound identification and reliable quantification in a single analytical run.
• To provide a rapid and high-confidence screening method for clinical toxicology laboratories.

Main Methods:

• Development of a fully automated sample preparation using a programmable liquid handler.
• Coupling of automated extraction with liquid chromatography-tandem mass spectrometry (LC-MS-MS).
• Utilized positive/negative ionization modes with up to 15 multiple reaction monitoring (MRM) transitions per compound for identification and quantification.

Main Results:

• The method successfully identified and quantified 245 compounds with high confidence using 15 MRM transitions per compound.
• Validated quantitative performance showed satisfactory intra- and inter-assay imprecision and inaccuracy (CV% < 20%).
• Excellent correlation (R² = 0.96) was observed when compared to dedicated quantitative methods using 175 sample pairs.

Conclusions:

• The developed automated LC-MS-MS screening procedure provides high confidence in compound detection for clinical toxicology.
• The method's speed (8 min sample prep + 10 min chromatography) and reliability make it suitable for core laboratories.
• This first-line screening approach aids in rapid and accurate diagnosis of drug overdose and poisoning.