title: Fully automated semi-quantitative toxicological screening in three biological matrices using turbulent flow chromatography/high resolution mass spectrometry.
abstract: BACKGROUND
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: CHU Clermont-Ferrand, Service de Pharmacologie Médicale, Laboratoire de pharmacologie-toxicologie, F-63003 Clermont-Ferrand, France; Inserm, UMR 1107 Neuro-Dol, F-63001 Clermont-Ferrand, France. Electronic address: l-roche@chu-clermontferrand.fr.
authors.forename: Lucie
authors.initials: L
authors.lastname: Roche
authors.affiliation: CHU Clermont-Ferrand, Service de Pharmacologie Médicale, Laboratoire de pharmacologie-toxicologie, F-63003 Clermont-Ferrand, France; Inserm, UMR 1107 Neuro-Dol, F-63001 Clermont-Ferrand, France.
authors.forename: Jérémy
authors.initials: J
authors.lastname: Pinguet
authors.affiliation: CHU Clermont-Ferrand, Service de Pharmacologie Médicale, Laboratoire de pharmacologie-toxicologie, F-63003 Clermont-Ferrand, France; Inserm, UMR 1107 Neuro-Dol, F-63001 Clermont-Ferrand, France.
authors.forename: Pauline
authors.initials: P
authors.lastname: Herviou
authors.affiliation: CHU Clermont-Ferrand, Service de Pharmacologie Médicale, Laboratoire de pharmacologie-toxicologie, F-63003 Clermont-Ferrand, France; Clermont Université, Université d'Auvergne, Laboratoire de Pharmacologie Médicale, Faculté de Médecine, F-63001 Clermont-Ferrand, France; Inserm, UMR 1107 Neuro-Dol, F-63001 Clermont-Ferrand, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Libert
authors.affiliation: CHU Clermont-Ferrand, Service de Pharmacologie Médicale, Laboratoire de pharmacologie-toxicologie, F-63003 Clermont-Ferrand, France; Clermont Université, Université d'Auvergne, Laboratoire de Pharmacologie Médicale, Faculté de Médecine, F-63001 Clermont-Ferrand, France; Inserm, UMR 1107 Neuro-Dol, F-63001 Clermont-Ferrand, France.
authors.forename: Chouki
authors.initials: C
authors.lastname: Chenaf
authors.affiliation: CHU Clermont-Ferrand, Service de Pharmacologie Médicale, Laboratoire de pharmacologie-toxicologie, F-63003 Clermont-Ferrand, France; Clermont Université, Université d'Auvergne, Laboratoire de Pharmacologie Médicale, Faculté de Médecine, F-63001 Clermont-Ferrand, France; Inserm, UMR 1107 Neuro-Dol, F-63001 Clermont-Ferrand, France.
authors.forename: Alain
authors.initials: A
authors.lastname: Eschalier
authors.affiliation: CHU Clermont-Ferrand, Service de Pharmacologie Médicale, Laboratoire de pharmacologie-toxicologie, F-63003 Clermont-Ferrand, France; Clermont Université, Université d'Auvergne, Laboratoire de Pharmacologie Médicale, Faculté de Médecine, F-63001 Clermont-Ferrand, France; Inserm, UMR 1107 Neuro-Dol, F-63001 Clermont-Ferrand, France; CHU Clermont-Ferrand, Centre Addictovigilance Auvergne, F-63003 Clermont-Ferrand, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Authier
authors.affiliation: CHU Clermont-Ferrand, Service de Pharmacologie Médicale, Laboratoire de pharmacologie-toxicologie, F-63003 Clermont-Ferrand, France; Inserm, UMR 1107 Neuro-Dol, F-63001 Clermont-Ferrand, France.
authors.forename: Damien
authors.initials: D
authors.lastname: Richard
mesh_terms: D001331:Automation; D002845:Chromatography / Q000379:methods*; D006801:Humans; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D018675:Toxicity Tests
keywords: Clinical toxicology; High resolution mass spectrometry; LC–MS; Orbitrap; Toxicological screening; Turbulent flow chromatography