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Clinical Chemistry 55: 2004-2018, 2009. First published October 1, 2009; 10.1373/clinchem.2008.122341
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(Clinical Chemistry. 2009;55:2004-2018.)
© 2009 American Association for Clinical Chemistry, Inc.

Drug Monitoring and Toxicology

Simultaneous Screening and Quantification of 29 Drugs of Abuse in Oral Fluid by Solid-Phase Extraction and Ultraperformance LC-MS/MS

Nora Badawi1, Kirsten Wiese Simonsen1, Anni Steentoft1, Inger Marie Bernhoft2 and Kristian Linnet1,a

1 Section of Forensic Chemistry, Department of Forensic Medicine, Faculty of Health Sciences, University of Copenhagen; 2 Department of Transport, Technical University of Denmark, Kgs. Lyngby, Denmark.

aAddress correspondence to this author at: Section of Forensic Chemistry, Department of Forensic Medicine, Faculty of Health Sciences, University of Copenhagen, Frederik V’s vej 11, 3. DK-2100, Denmark. Fax +45-3532-6085; e-mail kristian.linnet{at}forensic.ku.dk.

Background: The European DRUID (Driving under the Influence of Drugs, Alcohol And Medicines) project calls for analysis of oral fluid (OF) samples, collected randomly and anonymously at the roadside from drivers in Denmark throughout 2008–2009. To analyze these samples we developed an ultra performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) method for detection of 29 drugs and illicit compounds in OF. The drugs detected were opioids, amphetamines, cocaine, benzodiazepines, and {Delta}-9-tetrahydrocannabinol.

Method: Solid-phase extraction was performed with a Gilson ASPEC XL4 system equipped with Bond Elut Certify sample cartridges. OF samples (200 mg) diluted with 5 mL of ammonium acetate/methanol (vol/vol 90:10) buffer were applied to the columns and eluted with 3 mL of acetonitrile with aqueous ammonium hydroxide. Target drugs were quantified by use of a Waters ACQUITY UPLC system coupled to a Waters Quattro Premier XE triple quadrupole (positive electrospray ionization mode, multiple reaction monitoring mode).

Results: Extraction recoveries were 36%–114% for all analytes, including {Delta}-9-tetrahydrocannabinol and benzoylecgonine. The lower limit of quantification was 0.5 µg/kg for all analytes. Total imprecision (CV) was 5.9%–19.4%. With the use of deuterated internal standards for most compounds, the performance of the method was not influenced by matrix effects. A preliminary account of OF samples collected at the roadside showed the presence of amphetamine, cocaine, codeine, {Delta}-9-tetrahydrocannabinol, tramadol, and zopiclone.

Conclusions: The UPLC-MS/MS method makes it possible to detect all 29 analytes in 1 chromatographic run (15 min), including {Delta}-9-tetrahydrocannabinol and benzoylecgonine, which previously have been difficult to incorporate into multicomponent methods.






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