title: Biomonitoring method for bisphenol A in human urine by ultra-high-performance liquid chromatography-tandem mass spectrometry.
abstract: An ultra-high-performance liquid chromatography-tandem mass spectrometry method for the measurement of total bisphenol A in human urine was developed and validated. The method utilized liquid/liquid extraction with 1-chlorobutane and a human urine aliquot size of 800μL. Chromatography was performed on an Acquity UPLC(®) system with a Kinetex(®) Phenyl-Hexyl column. Mass spectrometric analysis was with negative electrospray ionization on a Quattro Premier XE™. The surrogate matrix method was used for the preparation of calibration standards in synthetic urine due to the presence of BPA in control human urine. The validated calibration range was 0.75-20ng/mL with a limit of detection of 0.1ng/mL. The internal standard was d16-bisphenol A. Method validation utilized quality control samples at three concentrations in both synthetic urine and human urine. Bisphenol A mono-glucuronide was fortified in synthetic urine in each analytical run to monitor the enzymatic conversion of the glucuronide conjugate to BPA by β-glucuronidase. Validated method parameters included linearity, accuracy, precision, integrity of dilution, selectivity, re-injection reproducibility, recovery/matrix effect, solution stability, and matrix stability in human urine. Acceptance criteria for analytical standards and QCs were ±20% of nominal concentration. Matrix stability in human urine was validated after 24h at ambient temperature, after three freeze/thaw cycles, and after frozen storage at -20°C and -80°C for up to 218 days. The method has been applied to the analysis of over 1750 human urine samples from a biomonitoring study. The median and mean urine BPA concentrations were 2.71ng/mL and 4.75ng/mL, respectively.
authors.affiliation: Center for Human Toxicology, University of Utah, 30 South 2000 East, Room 201, Salt Lake City, UT 84112, United States. Electronic address: anderson.dave@utah.edu.
authors.forename: David J
authors.initials: DJ
authors.lastname: Anderson
authors.affiliation: Center for Human Toxicology, University of Utah, 30 South 2000 East, Room 201, Salt Lake City, UT 84112, United States.
authors.forename: Eric M
authors.initials: EM
authors.lastname: Brozek
authors.affiliation: Department of Family and Preventive Medicine, Division of Public Health, University of Utah, 375 Chipeta Way, Suite A, Salt Lake City, UT 84108, United States.
authors.forename: Kyley J
authors.initials: KJ
authors.lastname: Cox
authors.affiliation: Department of Family and Preventive Medicine, Division of Public Health, University of Utah, 375 Chipeta Way, Suite A, Salt Lake City, UT 84108, United States.
authors.forename: Christina A
authors.initials: CA
authors.lastname: Porucznik
authors.affiliation: Center for Human Toxicology, University of Utah, 30 South 2000 East, Room 201, Salt Lake City, UT 84112, United States; Department of Pathology, University of Utah, 15 North Medical Drive, Salt Lake City, UT 84112, United States.
authors.forename: Diana G
authors.initials: DG
authors.lastname: Wilkins
mesh_terms: D001559:Benzhydryl Compounds / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D010636:Phenols / Q000652:urine*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D000098826:Bisphenol A Compounds
chemical_list: D001559:Benzhydryl Compounds; D010636:Phenols; C006780:bisphenol A; D000098826:Bisphenol A Compounds
keywords: Bisphenol A; Human urine; Liquid chromatography; Tandem mass spectrometry