title: Simultaneous Screening and Quantification of Basic, Neutral and Acidic Drugs in Blood Using UPLC-QTOF-MS.
abstract: An analytical method using ultra performance liquid chromatography (UPLC) quadrupole time-of-flight mass spectrometry (QTOF-MS) was developed and validated for the targeted toxicological screening and quantification of commonly used pharmaceuticals and drugs of abuse in postmortem blood using 100 µL sample. It screens for more than 185 drugs and metabolites and quantifies more than 90 drugs. The selected compounds include classes of pharmaceuticals and drugs of abuse such as: antidepressants, antipsychotics, analgesics (including narcotic analgesics), anti-inflammatory drugs, benzodiazepines, beta-blockers, amphetamines, new psychoactive substances (NPS), cocaine and metabolites. Compounds were extracted into acetonitrile using a salting-out assisted liquid-liquid extraction (SALLE) procedure. The extracts were analyzed using a Waters ACQUITY UPLC coupled with a XEVO QTOF mass spectrometer. Separation of the analytes was achieved by gradient elution using Waters ACQUITY HSS C18 column (2.1 mm x 150 mm, 1.8 μm). The mass spectrometer was operated in both positive and negative electrospray ionization modes. The high-resolution mass spectrometry (HRMS) data was acquired using a patented Waters MSE acquisition mode which collected low and high energy spectra alternatively during the same acquisition. Positive identification of target analytes was based on accurate mass measurements of the molecular ion, product ion, peak area ratio and retention times. Calibration curves were linear over the concentration range 0.05-2 mg/L for basic and neutral analytes and 0.1-6 mg/L for acidic analytes with the correlation coefficients (r2) > 0.96 for most analytes. The limits of detection (LOD) were between 0.001-0.05 mg/L for all analytes. Good recoveries were achieved ranging from 80% to 100% for most analytes using the SALLE method. The method was validated for sensitivity, selectivity, accuracy, precision, stability, carryover and matrix effects. The developed method was tested on a number of authentic forensic samples producing consistent results that correlated with results obtained from other validated methods.
authors.affiliation: Forensic Toxicology Laboratory, NSW Forensic & Analytical Science Service, Lidcombe, NSW, 2141, Australia.
authors.forename: Sergei
authors.initials: S
authors.lastname: Bidny
authors.affiliation: Forensic Toxicology Laboratory, NSW Forensic & Analytical Science Service, Lidcombe, NSW, 2141, Australia.
authors.forename: Kim
authors.initials: K
authors.lastname: Gago
authors.affiliation: Forensic Toxicology Laboratory, NSW Forensic & Analytical Science Service, Lidcombe, NSW, 2141, Australia.
authors.forename: Phuong
authors.initials: P
authors.lastname: Chung
authors.affiliation: Forensic Toxicology Laboratory, NSW Forensic & Analytical Science Service, Lidcombe, NSW, 2141, Australia.
authors.forename: Desdemona
authors.initials: D
authors.lastname: Albertyn
authors.affiliation: Centre for Forensic Science, University of Technology Sydney, Ultimo, NSW, 2007, Australia.
authors.forename: Daniel
authors.initials: D
authors.lastname: Pasin
mesh_terms: D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D053593:Forensic Toxicology / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D013287:Illicit Drugs / Q000097:blood / Q000737:chemistry; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction; D013058:Mass Spectrometry / Q000379:methods*; D004364:Pharmaceutical Preparations / Q000097:blood* / Q000737:chemistry; D012015:Reference Standards; D015203:Reproducibility of Results
chemical_list: D013287:Illicit Drugs; D004364:Pharmaceutical Preparations