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J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Feb 1;947-948:83-95. doi: 10.1016/j.jchromb.2013.12.014. Epub 2013 Dec 25.

Screening and quantitative determination of drugs of abuse in diluted urine by UPLC-MS/MS.

Author information

  • 1Department of Clinical Pharmacology, St. Olav University Hospital, Trondheim, Norway. Electronic address:
  • 2Waters Corporation Sweden AB, Sollentuna, Sweden.
  • 3Department of Clinical Pharmacology, St. Olav University Hospital, Trondheim, Norway; Department of Laboratory Medicine, Women's and Children's Health, Norwegian University of Science and Technology, Trondheim, Norway.
  • 4Department of Clinical Pharmacology, St. Olav University Hospital, Trondheim, Norway.


The purpose of this work was to develop and evaluate a fast, robust and specific UPLC-MS/MS screening platform for the determination and quantification of a variety of commonly used drugs of abuse in urine, i.e. a high-throughput quantitative analysis. Substances in the drug classes opioids, central nervous system stimulants and benzodiazepines and related agents were included in addition to cannabis and pregabalin, a total of 35 different analytes. Based on the concentrations and the physico-chemical properties of the substances, three UPLC-MS/MS methods were developed in parallel. Prior to analysis, sample preparation consisted of two different simple dilutions with 60 and 100 μL urine, respectively, using a Tecan Freedom Evo pipetting robot platform. A Waters Xevo TQ-S tandem quadrupole mass spectrometer coupled to a Waters I-class UPLC was used for quantitative analysis of one quantitative and one qualifying MRM transition for each analyte, except for tramadol for which the metabolite O-desmethyl-tramadol was included in the MRM method to confirm tramadol identity. Deuterated analogs were included as internal standards. The between-assay relative standard deviations varied from 2% to 11% and the limits of quantification were in the range 1-200 ng/mL for the various analytes. After development and initial testing, the method has been successfully implemented and routinely used at our hospital for quantitative screening of drugs of abuse in more than 35,000 urinary samples.

Copyright © 2013 Elsevier B.V. All rights reserved.


Benzodiazepines; Central nervous system stimulants; Drugs of abuse; Opioids; Screening; UPLC–MS/MS

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