title: Development of SPME-LC-MS method for screening of eight beta-blockers and bronchodilators in plasma and urine samples.
abstract: The current work describes the development and validation of a simple, efficient, and fast method using solid phase microextraction coupled to liquid chromatography-tandem mass spectrometry (SPME-LC-MS/MS) for the concomitant measurement of eight beta-blockers and bronchodilators in plasma and urine. The presented assay enables quantitative determination of acebutolol, atenolol, fenoterol, nadolol, pindolol, procaterol, sotalol, and timolol. In this work, samples were prepared on a high-throughput platform using the 96-well plate format of the thin film solid phase microextraction (TFME) system, and a biocompatible extraction phase made of hydrophilic-lipophilic balance particles. Analytes were separated on a pentafluorophenyl column (100mm×2.1mm, 3μm) by gradient elution using an UPLC Nexera coupled with an LCMS-8060 mass spectrometer. The mobile phase consisted of water-acetonitrile (0.1% formic acid) at a flow rate of 0.4mLmin(-1). The linearity of the method was checked within therapeutic blood-plasma concentrations, and shown to adequately reflect typically expected concentrations of future study samples. Post-extraction addition experiments showed that the matrix effect ranged in plasma from 98% for procaterol to 115% for nadolol, and in urine, from 85% for nadolol and pindolol to 119% for atenolol. The method was successfully validated using Food and Drug Administration (FDA) guidelines, and met all acceptance criteria for bioanalytical assays at five concentration levels for all selected drugs. The final protocol can be successfully applied for monitoring concentrations of the selected drugs in both plasma and urine matrices obtained from patients or athletes.
authors.affiliation: Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toruń, Toruń, Poland. Electronic address: gorynski@cm.umk.pl.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Goryński
authors.affiliation: Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toruń, Toruń, Poland.
authors.forename: Alicja
authors.initials: A
authors.lastname: Kiedrowicz
authors.affiliation: Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toruń, Toruń, Poland. Electronic address: bbojko@cm.umk.pl.
authors.forename: Barbara
authors.initials: B
authors.lastname: Bojko
mesh_terms: D000319:Adrenergic beta-Antagonists / Q000097:blood / Q000652:urine; D001993:Bronchodilator Agents / Q000097:blood / Q000652:urine; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D004300:Doping in Sports / Q000379:methods; D016903:Drug Monitoring / Q000379:methods; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D015394:Molecular Structure; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052617:Solid Phase Microextraction / Q000379:methods*
chemical_list: D000319:Adrenergic beta-Antagonists; D001993:Bronchodilator Agents
keywords: Beta-blockers; High throughput; Liquid chromatography–mass spectrometry; Plasma; SPME; Sample preparation; Solid phase microextraction; Urine