Abstract

In clinical and forensic toxicology, fast and specific methods are needed for the screening of different classes of drugs. A complete general unknown screening procedure was developed using turbulent flow chromatography with electrospray ionization and Orbitrap mass spectrometry.After protein precipitation, samples were injected directly into the turbulent flow chromatographic system and analyzed with an Orbitrap mass spectrometer. The Exactive® operated in positive and negative modes with alternated high collision dissociation in order to obtain characteristic fragments. We built a library containing 616 compounds by analyzing a reference standard for all the molecules.Identification was based on retention time, accurate measured mass, isotopic pattern and presence of specific fragments. For each substance, we set a calibration range encompassing infra-therapeutic, therapeutic, supra-therapeutic and toxic concentrations in order to generate semi-quantitative result. For 65% of the components, the limit of detection was below 5 ng/mL. The validation process showed the approach to be selective, sensitive, accurate and precise.The method has been accredited by COFRAC (French Accreditation Committee) according to the ISO 15189 standard. Applicability was successfully tested by analyzing authentic serum, urine and whole blood samples.

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