title: The determination of budesonide and fluticasone in human sputum samples collected from COPD patients using LC-MS/MS.
abstract: A bioanalytical method for the quantitative determination of budesonide and fluticasone in human sputum was developed. Sputolysin(®) Reagent was added to the sputum samples. After incubation (37°C; 60-70 min under shaking) and automated solid phase extraction the extracts were analysed using LC-MS/MS. Budesonide and fluticasone showed good linearity (r>0.99) over the range 0.1-100 nM in the first and second validation batch, and over the range 0.25-10,000 nM in the third and fourth validation batch. The lower limit of quantification (LLOQ) achieved was 5 nM for budesonide and fluticasone in 100 μL human sputum. Intra-run and inter-run RSD for four quality control levels (5-100 nM) were within 6.9% (budesonide) and 8.0% (fluticasone). The accuracy ranged from -11.4% to -1.6% (budesonide), and from -11.8% to 0.4% (fluticasone). The validated method was applied to clinical sputum samples from COPD patients.
authors.affiliation: TNO Triskelion BV, Analytical Research Department, Zeist, The Netherlands. brigitte.buscher@tno.nl
authors.forename: B A P
authors.initials: BA
authors.lastname: Buscher
authors.forename: H
authors.initials: H
authors.lastname: Jägfeldt
authors.forename: H
authors.initials: H
authors.lastname: Sandman
authors.forename: R
authors.initials: R
authors.lastname: Brust-van Schaik
authors.forename: F
authors.initials: F
authors.lastname: van Schaik
authors.forename: L P
authors.initials: LP
authors.lastname: Brüll
mesh_terms: D000730:Androstadienes / Q000032:analysis* / Q000493:pharmacokinetics / Q000627:therapeutic use; D000893:Anti-Inflammatory Agents / Q000032:analysis / Q000493:pharmacokinetics / Q000627:therapeutic use; D019819:Budesonide / Q000032:analysis* / Q000493:pharmacokinetics / Q000627:therapeutic use; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D000068298:Fluticasone; D006801:Humans; D016014:Linear Models; D029424:Pulmonary Disease, Chronic Obstructive / Q000188:drug therapy* / Q000378:metabolism*; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D013183:Sputum / Q000737:chemistry* / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013696:Temperature
chemical_list: D000730:Androstadienes; D000893:Anti-Inflammatory Agents; D019819:Budesonide; D000068298:Fluticasone