Liquid chromatography tandem mass spectrometry determination of total budesonide levels in dog plasma after inhalation exposure.
ABSTRACT A sensitive and selective method to quantify budesonide in dog plasma samples was developed and fully validated. Liquid-liquid extraction was followed by solid-phase extraction and liquid chromatography-tandem mass spectrometry with electrospray ionization. After reconstitution of the analytes in the mobile phase, samples were analysed by reversed-phase liquid chromatography with isocratic elution. d8-Budesonide was used as an internal standard, and characteristic transitions of d8-budesonide and budesonide were used for quantification. The method was validated with respect to selectivity, specificity, linearity, recovery, repeatability, reproducibility and limits of detection and quantification. The validated method was successfully applied to monitor the plasma levels of budesonide in dogs exposed to clinical doses of inhaled and intravenous drug.
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ABSTRACT: Budesonide (BUD) is used as a mixture of 22R and 22S epimers for the topical treatment of asthma, rhinitis, and inflammatory bowel disease. To study stereoselectivity in the pharmacokinetics of each epimer, we developed a stereoselective and sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry method for the quantitative determination of 22R and 22S epimers of BUD in human plasma. The epimers of BUD were extracted from plasma using n-hexane/dichloromethane/isopropanol (2:1:0.1, v/v/v) under alkaline conditions. Baseline separation was obtained within 7min on an Acquity UPLC BEH C (50mm×2.1mm, 1.7μm) column using an isocratic mobile phase consisting of acetonitrile/5mM ammonium acetate/acetic acid (29:71:0.142, v/v/v) at a flow rate of 0.7mL/min. Mass spectrometric detection was performed in a multiple reaction monitoring mode using the m/z 489→357 transition for BUD epimers and the m/z 497→357 transition for the internal standard d-BUD epimers. Calibration curves were linear over the concentration ranges of 5.0-500 and 5.0-3000pg/mL for 22R-BUD and 22S-BUD, respectively. The lower limit of quantification was 5.0pg/mL for both epimers. The method was successfully applied in a pharmacokinetic study of BUD controlled-release capsules in humans. Consistent differences in the pharmacokinetics of the 22R and 22S epimers were observed, the AUC of 22S-BUD was approximately six times higher than that of 22R-BUD, and the 22S-/22R-BUD ratio of total body clearance was 0.17.Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 03/2013; 921-922:27-34. · 2.78 Impact Factor