title: Quantitative analysis of unconjugated and total bisphenol A in human urine using solid-phase extraction and UPLC-MS/MS: method implementation, method qualification and troubleshooting.
abstract: The aim of the presented investigation was to document challenges encountered during implementation and qualification of a method for bisphenol A (BPA) analysis and to develop and discuss precautions taken to avoid and to monitor contamination with BPA during sample handling and analysis. Previously developed and published HPLC-MS/MS methods for the determination of unconjugated BPA (Markham et al. Journal of Analytical Toxicology, 34 (2010) 293-303) [17] and total BPA (Markham et al. Journal of Analytical Toxicology, 38 (2014) 194-203) [20] in human urine were combined and transferred into another laboratory. The initial method for unconjugated BPA was developed and evaluated in two independent laboratories simultaneously. The second method for total BPA was developed and evaluated in one of these laboratories to conserve resources. Accurate analysis of BPA at sub-ppb levels is a challenging task as BPA is a widely used material and is ubiquitous in the environment at trace concentrations. Propensity for contamination of biological samples with BPA is reported in the literature during sample collection, storage, and/or analysis. Contamination by trace levels of BPA is so pervasive that even with extraordinary care, it is difficult to completely exclude the introduction of BPA into biological samples and, consequently, contamination might have an impact on BPA biomonitoring data. The applied UPLC-MS/MS method was calibrated from 0.05 to 25ng/ml. The limit of quantification was 0.1ng/ml for unconjugated BPA and 0.2ng/ml for total BPA, respectively, in human urine. Finally, the method was applied to urine samples derived from 20 volunteers. Overall, BPA can be analyzed in human urine with acceptable recovery and repeatability if sufficient measures are taken to avoid contamination throughout the procedure from sample collection until UPLC-MS/MS analysis.
authors.affiliation: TNO Triskelion BV, Utrechtseweg 48, Zeist, The Netherlands.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Buscher
authors.affiliation: TNO Triskelion BV, Utrechtseweg 48, Zeist, The Netherlands.
authors.forename: Dick
authors.initials: D
authors.lastname: van de Lagemaat
authors.affiliation: Currenta GmbH & Co., OHG, Leverkusen, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Gries
authors.affiliation: Bayer Pharma Aktiengesellschaft, Wuppertal, Germany. Electronic address: dieter.beyer@bayer.com.
authors.forename: Dieter
authors.initials: D
authors.lastname: Beyer
authors.affiliation: The Dow Chemical Company, Midland, MI, USA.
authors.forename: Dan A
authors.initials: DA
authors.lastname: Markham
authors.affiliation: The Dow Chemical Company, Midland, MI, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Budinsky
authors.affiliation: Saudi Basic Industries Corporation (SABIC), Pittsfield, MA, USA.
authors.forename: Stephen S
authors.initials: SS
authors.lastname: Dimond
authors.affiliation: SABIC Research & Technology Pvt., Ltd., Bangalore, India.
authors.forename: Rajesh V
authors.initials: RV
authors.lastname: Nath
authors.affiliation: Covestro LLC, Pittsburgh, PA, USA.
authors.forename: Stephanie A
authors.initials: SA
authors.lastname: Snyder
authors.affiliation: American Chemistry Council, Washington DC, USA.
authors.forename: Steven G
authors.initials: SG
authors.lastname: Hentges
mesh_terms: D001559:Benzhydryl Compounds / Q000737:chemistry* / Q000302:isolation & purification / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D016014:Linear Models; D010636:Phenols / Q000737:chemistry* / Q000302:isolation & purification / Q000652:urine*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D000098826:Bisphenol A Compounds
chemical_list: D001559:Benzhydryl Compounds; D010636:Phenols; C006780:bisphenol A; D000098826:Bisphenol A Compounds