Title

A Dilute-and-Shoot Flow-Injection Tandem Mass Spectrometry Method for Quantification of Phenobarbital in Urine

Document Type

Article

Publication Date

1-15-2017

Publication Title

Rapid Communications in Mass Spectrometry

Abstract

RATIONALE: Liquid chromatography/tandem mass spectrometry (LC/MS/MS) is the gold standard of urine drug testing. However, current LC-based methods are time consuming, limiting the throughput of MS-based testing and increasing the cost. This is particularly problematic for quantification of drugs such as phenobarbital, which is often analyzed in a separate run because they must be negatively ionized.

METHODS: This study examined the feasibility of using a dilute-and-shoot flow-injection method without LC separation to quantify drugs with phenobarbital as a model system. Briefly, a urine sample containing phenobarbital was first diluted by 10 times, followed by flow injection of the diluted sample to mass spectrometer. Quantification and detection of phenobarbital were achieved by an electrospray negative ionization MS/MS system operated in the multiple reaction monitoring (MRM) mode with the stable-isotope-labeled drug as internal standard.

RESULTS: The dilute-and-shoot flow-injection method developed was linear with a dynamic range of 50–2000 ng/mL of phenobarbital and correlation coefficient > 0.9996. The coefficients of variation and relative errors for intra- and interassays at four quality control (QC) levels (50, 125, 445 and 1600 ng/mL) were 3.0% and 5.0%, respectively. The total run time to quantify one sample was 2 min, and the sensitivity and specificity of the method did not deteriorate even after 1200 consecutive injections.

CONCLUSIONS: Our method can accurately and robustly quantify phenobarbital in urine without LC separation. Because of its 2 min run time, the method can process 720 samples per day. This feasibility study shows that the dilute-and-shoot flow-injection method can be a general way for fast analysis of drugs in urine. Copyright © 2016 John Wiley & Sons, Ltd.

Comments

We acknowledge Cleveland State University for financially supporting this research and also thank “The National Science Foundation Major Research Instrumentation Grant” (CHE- 0923398) for supporting the requisition of the AB Sciex QTrap 5500 mass spectrometer instrument for this work.

DOI

10.1002/rcm.7763

Volume

31

Issue

1