Simultaneous analysis of glucocorticosteroid fluticasone propionate and its metabolite fluticasone propionate 17β-carboxylic acid in human plasma by UPLC–MS/MS at sub pg/mL level

    loading  Checking for direct PDF access through Ovid


A highly sensitive and rapid ultra performance liquid chromatography-tandem mass spectrometry method has been developed for the simultaneous determination of fluticasone propionate (FP) and its major metabolite, fluticasone propionate-17beta-carboxylic acid (FP 17β-CA) in human plasma. The analytes and their deuterated internal standards, FP-d3 and FP 17β-CA-d3 were extracted from 500 μL plasma samples by solid phase extraction on Oasis MAX cartridges. The chromatographic analysis was performed on ACQUITY UPLC BEH C18 (50 mm × 2.1 mm, 1.7 μm) column using methanol-acetonitrile (50:50, v/v) and 2.0 mM ammonium trifluroacetate (ATFA) (85:15, v/v) as the mobile phase. Following separation of the analytes, protonated precursor → product ion transitions (FP: m/z 501.1 → 293.2, FP17β-CA: m/z 453.3 → 293.2, FP-d3: m/z 504.2 → 293.2, FP 17β-CA-d3: m/z 456.3 → 293.2) were monitored on FP 17β-CA a triple quadrupole mass spectrometer, operating in multiple reaction monitoring (MRM) and positive ionization mode. The calibration curves were established in the range of 0.5–100 pg/mL with a correlation coefficient (r2) ≥ 0.9992 for both the analytes. The intra-batch and inter-batch accuracy and precision varied from 95.5-103.4% and 0.74-5.06% across quality controls for both the analytes. The mean assay recoveries for FP and FP 17β-CA were 84.2% and 93.5% respectively. The validated method was successfully applied to support a bioequivalence study of 200 μg FP, administered using nasal spray formulation in 18 healthy Indian subjects. Reproducibility of the method was assessed by reanalysis of 98 incurred study samples.

Related Topics

    loading  Loading Related Articles