title: 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.
abstract: 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 500μL plasma samples by solid phase extraction on Oasis MAX cartridges. The chromatographic analysis was performed on ACQUITY UPLC BEH C18 (50mm×2.1mm, 1.7μm) column using methanol-acetonitrile (50:50, v/v) and 2.0mM 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 0.5-100pg/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 200μg FP, administered using nasal spray formulation in 18 healthy Indian subjects. Reproducibility of the method was assessed by reanalysis of 98 incurred study samples.
authors.affiliation: Department of Chemistry, School of Sciences, Gujarat University, Navrangpura, Ahmedabad 380009, Gujarat, India.
authors.forename: Sneha G
authors.initials: SG
authors.lastname: Nair
authors.affiliation: Laboratory of Metabolism, Center for Cancer Research, National Cancer Institute, Bethesda, MD 20892, United States.
authors.forename: Daxesh P
authors.initials: DP
authors.lastname: Patel
authors.affiliation: Department of Chemistry, St. Xavier's College, Navrangpura, Ahmedabad 380009, Gujarat, India.
authors.forename: Mallika
authors.initials: M
authors.lastname: Sanyal
authors.affiliation: Bioanalytical Department, Alkem Laboratories Ltd., Lower Parel, Mumbai 400013, India.
authors.forename: Puran
authors.initials: P
authors.lastname: Singhal
authors.affiliation: Department of Chemistry, School of Sciences, Gujarat University, Navrangpura, Ahmedabad 380009, Gujarat, India. Electronic address: pranav_shrivastav@yahoo.com.
authors.forename: Pranav S
authors.initials: PS
authors.lastname: Shrivastav
mesh_terms: D000305:Adrenal Cortex Hormones / Q000097:blood / Q000378:metabolism; D000730:Androstadienes / Q000097:blood* / Q000378:metabolism; D002138:Calibration / Q000592:standards; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000592:standards; D000068298:Fluticasone / Q000097:blood* / Q000378:metabolism; D005938:Glucocorticoids / Q000097:blood* / Q000378:metabolism; D006801:Humans; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards
chemical_list: D000305:Adrenal Cortex Hormones; D000730:Androstadienes; D005938:Glucocorticoids; C549234:fluticasone propionate-17-carboxylic acid; D000068298:Fluticasone
keywords: Fluticasone propionate; Fluticasone propionate 17β-carboxylic acid; Human plasma; Nasal spray; Sub pg/mL; UPLC–MS/MS