Sneha G. Nair, Daxesh P. Patel, M. Sanyal
Feb 20, 2017
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Influential Citations
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Journal
Journal of Pharmaceutical and Biomedical Analysis
Abstract
&NA; 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 17&bgr;‐CA) in human plasma. The analytes and their deuterated internal standards, FP‐d3 and FP 17&bgr;‐CA‐d3 were extracted from 500 &mgr;L 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 &mgr;m) 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, FP17&bgr;‐CA: m/z 453.3 → 293.2, FP‐d3: m/z 504.2 → 293.2, FP 17&bgr;‐CA‐d3: m/z 456.3 → 293.2) were monitored on FP 17&bgr;‐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 17&bgr;‐CA were 84.2% and 93.5% respectively. The validated method was successfully applied to support a bioequivalence study of 200 &mgr;g FP, administered using nasal spray formulation in 18 healthy Indian subjects. Reproducibility of the method was assessed by reanalysis of 98 incurred study samples. Graphical abstract Figure. No caption available. HighlightsSub pg/mL level estimation of fluticasone propionate(FP) and FP 17&bgr;‐carboxylic acid.Simultaneous extraction of analytes from human plasma using SPE on MAX cartridge.Almost complete absence of FP 17&bgr;‐carboxylic acid in plasma samples of subjects.Bioequivalence study with 200 &mgr;g FP after nasal administration in healthy subjects.Method reproducibility is established by reanalysis of 98 incurred samples.