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  • Title: Validated assay for the simultaneous determination of cortisol and budesonide in human plasma using ultra high performance liquid chromatography-tandem mass spectrometry.
    Author: Szeitz A, Manji J, Riggs KW, Thamboo A, Javer AR.
    Journal: J Pharm Biomed Anal; 2014 Mar; 90():198-206. PubMed ID: 24389462.
    Abstract:
    An ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC/MS/MS) method was developed and validated for the quantification of cortisol and budesonide in human plasma. Charcoal stripped human plasma was used as the blank matrix during validation. Cortisol, budesonide, and dexamethasone (internal standard) were extracted from human plasma with methyl-tert-butyl ether, and the chromatographic separation of the peaks was achieved using a Waters Acquity UPLC BEH C18, 1.7 μm, 2.1 mm × 50 mm column with a run time of 4.0 min. Cortisol, budesonide, and dexamethasone were monitored at the total ion current of their respective multiple reaction monitoring transition signals. The UHPLC/MS/MS system consisted of an Agilent 1290 Infinity ultra high performance liquid chromatograph coupled with an AB Sciex Qtrap(®) 5500 hybrid linear ion-trap triple quadrupole mass spectrometer. The method was validated for accuracy, precision, linearity, range, selectivity, lower limit of quantification (LLOQ), recovery, matrix effect, dilution integrity, and evaluation of carry-over. All validation parameters met the acceptance criteria according to regulatory guidelines. The LLOQ was 1.0 ng/mL for both compounds requiring 100 μL of sample. To our knowledge, this is the first validated LC/MS/MS method for the simultaneous quantitative analysis of cortisol and budesonide in human plasma. The method was applied successfully in a clinical investigation of the impact of nasally administered Pulmicort (budesonide) on the hypothalamic-pituitary-adrenal axis of patients with chronic rhinosinusitis.
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