title: Determination of bronchoalveolar lavage bile acids by solid phase microextraction liquid chromatography-tandem mass spectrometry in combination with metabolite profiling: comparison with enzymatic assay.
abstract: A thin-film solid-phase microextraction (SPME) method coupled to liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis was developed for high-throughput determination of bronchoalveolar lavage bile acids. The proposed method was validated according to the bioanalytical method validation guidelines. LOQ and LLOQ were 0.007 and 0.02 μmol/L, respectively. The accuracy and the precision were <7 and <4%, respectively. The performance of the proposed method was also compared with an optimized enzymatic cycling assay. Results showed a weak correlation between total concentration of bile acid (BAs) obtained with enzymatic assay and cumulative concentration of specific BAs determined with SPME-LC-MS/MS. This discrepancy was probably due to the presence of other BAs in bronchoalveolar lavage fluid (BALF) samples. Metabolites profiling of BALF samples extracts using a high-resolution mass spectrometer (HRMS) revealed the presence of additional BAs, which were not included in the proposed method. After considering these additional BAs, a strong correlation was found between the LC-MS method and the enzymatic assay. Unsupervised statistical analysis conducted on HRMS data also showed clear separation within BALF samples, depending on the presence of BAs and other lipids. SPME-LC-MS-based metabolites profiling may provide additional information for diagnosis occurrence and severity of gastric reflux/aspiration in lung transplant patients.
authors.affiliation: Department of Chemistry, University of Waterloo, Waterloo, Ontario N2L 3G1 Canada.
authors.forename: Vincent
authors.initials: V
authors.lastname: Bessonneau
authors.affiliation: Department of Chemistry, University of Waterloo, Waterloo, Ontario N2L 3G1 Canada; Department of Medicinal Chemistry, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University, Toruń, Poland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Bojko
authors.affiliation: Mount Sinai Services Inc., Toronto, Ontario M5G 1Z5, Canada.
authors.forename: Azar
authors.initials: A
authors.lastname: Azad
authors.affiliation: Division of Thoracic Surgery, Toronto General Hospital, University Health Network, Toronto, Ontario M5G 2C4, Canada.
authors.forename: Shalf
authors.initials: S
authors.lastname: Keshavjee
authors.affiliation: Division of Thoracic Surgery, Toronto General Hospital, University Health Network, Toronto, Ontario M5G 2C4, Canada.
authors.forename: Sassan
authors.initials: S
authors.lastname: Azad
authors.affiliation: Department of Chemistry, University of Waterloo, Waterloo, Ontario N2L 3G1 Canada. Electronic address: janusz@uwaterloo.ca.
authors.forename: Janusz
authors.initials: J
authors.lastname: Pawliszyn
mesh_terms: D001647:Bile Acids and Salts / Q000032:analysis* / Q000378:metabolism; D018893:Bronchoalveolar Lavage; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods*; D006801:Humans; D015203:Reproducibility of Results; D052617:Solid Phase Microextraction / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D001647:Bile Acids and Salts
keywords: Bile acids; Bronchoalveolar lavage; Enzymatic assay; LC-MS; Metabolomics; SPME