A simple validated multi‐analyte method for detecting drugs in oral fluid by ultra‐performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS)

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EarlyView Article

  • Published: Dec 18, 2017
  • Author: Yufang Zheng, Erik Sparve, Mats Bergström
  • Journal: Drug Testing and Analysis


Oral fluid sampling offers several advantages compared to other methods of detecting drugs in biological matrices due to its easy sampling procedure and simple supervision. Hence, the use of oral fluid for drug testing is increasing in workplaces and in roadside testing. As a result, more laboratories are required to perform analyses of drugs in oral fluid.

A multi‐method using ultra performance liquid chromatographytandem mass spectrometry (UPLCMS/MS) for tracing drugs in oral fluid was developed. Oral fluid was collected using the collection device Oral‐Eze. Samples were prepared by dilution with acetonitrile and methanol followed by centrifugation prior to UPLCMS/MS analysis. The UPLC separation was achieved by using a BEH‐C18 column. Mass detection was performed in positive ion mode, and the most common drugs/metabolites were detected for amphetamines, benzodiazepines, cocaine, benzoylecgonine, opiates, opioids, phencyclidine, and Δ9‐ tetrahydrocannabinol.

Thirty‐seven analytes were validated using the developed UPLCMS/MS multi‐analyte method with a total run time of 5 minutes. Calibration curves were linear for all analytes over the concentration range 1.5600 ng/mL. Within‐ and between‐day CVs varied from 1.2% to 20% and from 0.7% to 20%, respectively, for most substances. Extraction recoveries from the oral fluid device were >70%, a few had a yield of 50%.

The developed multi‐method was successfully validated and applied in a clinical routine laboratory to detect drugs in oral fluid samples that were sent from different workplaces and clinics. High sensitivity, simple sample pretreatment and short analysis time are advantages of this method.

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